Datasets:

farleyknight

/

big_patent_10_percent

like 1

the present invention preferably comprises a mixture of natural products having anti - inflammatory , anti - irritant and wound healing properties , ( hereinafter “ active ingredients ”) in a polyhydroxy solvent wherein said solvent is a mixture of one or more glycol compounds or dihuydric alcohol compounds and one or more glycerin compounds , or trihydric alcohol compounds e . g . a diglycerine . the active ingredients may comprise from about 0 . 5 to about 10 %, by weight , of said ear drop composition . more preferably , said active ingredients may comprise from about 0 . 75 to about 5 %, by weight , e . g . from about 1 to about 3 %, by weight , of said ear drop composition . the glycol compounds may comprise dihydroxyalkyl compounds , such as 1 , 3 butylenegylcol , ethylene glycol , 1 , 2 propylene glycol or hydroxyalkyloxyalkyl compounds such hydroxymethyleneoxyethylene glycol , etc . most preferably , said glycol is 1 , 3 butyleneglycol , which besides having the necessary solvent properties provides anti - fungal and anti - bacterial properties . the glycerin compound or trihydric alcohol may comprise glycerin from natural or synthetic sources , preferably from natural sources . in addition , to functioning as a solvent in the eardrop compositions of this invention glycerin provides emollient , humectant , and lubrication properties to the present compositions . an anti - irritant agent derived from a natural source , e . g . an avenanthramide wherein m is an integer , e . g . of from 1 to 4 , i . e . 1 , 2 or 3 . a preferred avenanthramide source is a product sold as drago - calm available from syrmise of teterboro , new jersey , 00768 . drago - calm is reported to be an oat ( avena satvia ) kernel extract having a guaranteed active constituent concentration of 100 ppm avenanthramides this product has superior anti - irritant and anti - oxidant properties . the avenanthramides include in this product are analogues of tranilast ™, which is a potent anti - histamine the anti - irritant agent may comprise from about 0 . 1 to about 5 %, by weight , preferably from 0 . 5 to about 1 . 5 %, by weight , of the eardrop composition of this invention . the composition of this invention may further comprise one or more compounds having natural wound healing activity . preferably , said wound healing compounds may comprise aloe and / or alpha - bisbolol . aloe is a gel obtained by crushing the mucilaginous cells found in the inner tissue of the plant leaf . aloe is a wound healing agent and has emollient properties . the emollient effect of this gel is largely attributed to a constituent polysaccharide ( glucomannan ) similar to guar gum . the term “ aloe ” refers to a solid residue obtained by evaporating the latex derived from the outer layers of the plant leaf . aloe latex also contains the anthraquinone barbaloin ( a glucoside of aloe - emodin ). topical application of aloe in the compositions of this invention alleviates irritation , inhibit infection and promote healing of abrasions , minor wounds , and bums . the beneficial effects of aloe gel in relieving skin irritation in dry ear may be due , in part , to its moisturizing activity which reduces drying of abraded or injured skin . some clinical studies have demonstrated acceleration of wound healing from aloe . alpha bisabolol or (+) epi - alpha - bisobolol is a terpenoid which is the wound healing principal of peperomia galioides and also has anti - inflammatory properties . natural alpha bisabolol is available from basf . the wound healing components of the present eardrop compositions , i . e . aloe and / or alpha bisabolol , may comprise from about 0 . 01 to about 1 %, by weight , preferably from about 0 . 05 to about 0 . 15 %, by weight . the compositions of the present invention preferably may comprise further anti - inflammatory agents , which agents may , in addition , have anti - allergy and analgesic properties such as salts of glycyrrhic acid , e . g . dipotassium glycyrrhizate and white willow bark extract , respectively . the bark of the white willow tree is a source of salicin and other salicylic — compounds which are similar in structure to aspirin ( acetyl salicylic acid ) native americans are thought to have used ground willow bark a as a medicinal remedy for everything from pain relief to fevers . today , white willow bark is often used as a natural alternative aspirin — one of the most common uses in dietary supplements is as a adjunct for weight loss . thus , the extract of white willow tree bark is utilized in the present compositions as a pain reliever ( headaches , arthritis , minor injuries ) and an anti - inflammatory agent . the primary active compound in white willow bark is salicin . in the body salicin can be converted into salicylic acid , which has powerful effects as an anti - inflammatory and pain reliever . the anti - inflammatory agents may comprise from about 0 . 001 to about 1 %, by weight of the compositions of this invention , e . g . from about 0 . 002 to about 0 . 05 %, by weight , of such compositions . the compositions of the present invention may further comprise additional humectants in addition to glycerin discussed above . preferably , the ear drop composition will comprise a natural humectant such as pyrrolidone carboxylic acid , such as ajidew n - 50 which is the sodium salt of said acid and is available from ajinomoto usa , inc . of paramus , n . j . 07652 . finally , the compositions of the present invention may include one or more surfactants to stabilize the compositions . preferably , at least one of the surfactants is a cationic surfactant , e . g . a quatenary amine , such as stepanquat 50 nf which is a dialkyl dimethylammonium chloride and is available from stepan company . more preferably , the cationic surfactant stabilized compositions additionally include a nonionic surfactant such as tween 20 . the surfactants may comprise from about 0 . 02 to about 1 % by weight of said compositions , more preferably from about 0 . 2 to about 0 . 45 % by weight . the composition below discloses a preferred embodiment of this invention and the sources of the individual components . table percentage formula % by weight weight range 1 , 3 - butylene glycol 47 . 8300 % 40 - 50 diglycerine 801 50 . 0000 40 - 50 dragocalm # 2 / 060910 1 . 0000 0 . 5 - 1 . 5 ajidew n - 50 0 . 5000 0 . 25 - 0 . 75 abs white willow bar extract , powder 0 . 0100 . 005 -. 015 oristar dpg 0 . 2000 0 . 1 - 0 . 25 stepanquat 50 nf 0 . 1000 0 . 05 - 0 . 15 alpha - bisbolol , natural 0 . 1000 0 . 05 - 0 . 15 tween 20 0 . 2500 0 . 15 - 0 . 30 activaloe , # aa1210a 0 . 0100 0 . 005 -. 015 100 . 0000 % compounding formula batch size : 1 , 000 lbs . by weight part a 1 , 3 - butylene glycol 458 . 3000 lbs . diglycerine 801 500 . 000 drgocalm # 2 / 060910 10 . 000 ajidew n - 50 5 . 000 part b 1 , 3 - butylene glycol 20 . 000 abs white willow bark extract , powder 0 . 1000 oristar dpg 2 . 0000 stepanquat 50 nf 1 . 0000 alpha - bisbolol , natural 1 . 0000 tween 20 2 . 5000 activaloe , # aa1210a 0 . 1000 1 , 000 . 0000 lbs . in a separate tank , heat part b ingredients to 45 ° c . mix until all the solids are dissolved and the batch is uniform . add part b to part a . mix until uniform . sample for qc check . 1 . dd is a 56 caucasian female who has had recurrent external ear infections . she has been treated with various antibacterial and antifungal preparations . after finally getting the infections cleared , she has had no cerumen production in the external canal . with the use of the emollient and lubricating drops , she has successfully remained infection free for greater than six months while continuing her water activity . 2 . ah is a 12 year old asian american female with sensorineural hearing loss with a history of very dry ear canals and external ear infections . despite multiple medications it was difficult to clear her ear canals for long periods of time so that she could wear her hearing aid . once the infections had cleared with the use of the , emollient and lubricating drops , she has been able to successfully wear her hearing aids and follow up visits only every three months .

a method of treating diseases and conditions associated with dry ear comprising topically applying to the ear canal of a subject that is suffering therefrom a liquid composition comprising an anti - irritant agent , a wound healing agent ; and an anti - inflammatory agent dissolved in a polyhydroxy liquid solvent .

fig1 shows a stair climbing apparatus 5 comprised of a frame 10 on which is mounted a treadmill or endless belt 20 that has a rotation axis ax . the treadmill or belt 20 is mounted for rotation in a backwardly downwardly direction bd on pulleys 30 that drive the rotation of the treadmill 20 via gear teeth 32 adapted to mesh with complementary apertures in the links of chains , belts or the like 40 that support the individual forward link 50 and rearward link 60 of each stair via a rod 45 that extends between the drive chains 40 , belts or the like that are meshed or driven by front and rear pulleys 30 f , 30 r . the angle of incline a of the axis ax is selectively adjustable by the user 7 to any desired angle a relative to horizontal by conventional devices such as by way of a controller 14 having a manually operable user interface 14 i , the controller 14 being interconnected to and containing instructions that controllably drive a motor or other drive device 12 that is mechanically interconnected to the treadmill assembly in a manner that controllably moves the front axle 38 f or pulley 30 f in a generally up and down ud direction relative to the rear axle 38 r and rear pulley 30 r . most preferably the instructions contained within the controller 14 enable the user 7 to adjust the angle a of the longitudinal axis ax to an angle that enable the user to perform a stepping exercise or motion or action as shown in fig1 such that the longitudinal axis la of the user &# 39 ; s upper torso 9 is disposed in a generally vertical v orientation when the user &# 39 ; s upper torso or spine is disposed in a generally straight orientation as shown in fig1 . preferably the instructions contained within the controller 14 enable the longitudinal axis ax to be adjusted to an angle a of between about 30 and about 36 degrees , most preferably between about 32 and about 34 degrees . beginning from a start exercise motionless position of the treadmill or belt 20 at which point in time the treadmill is motionless , the treadmill 20 is held in a motionless position by an on - off brake 102 that prevents rotational movement of the pulleys 38 r , 38 f and the treadmill 20 until the on - off brake 102 is released by the user 7 . upon release of the on - off brake 102 , movement of the treadmill or belt 20 is preferably resisted during a run - up from 0 rotational speed to x in rotational speed , x being a preselected rotational speed greater than zero , solely by drag , friction and weight of the moving mechanical components of the apparatus such as the treadmill 20 , pulleys 30 f , 30 r , chains or belts 40 and associated components such as bearings and the like . starting from the motionless position , the treadmill 20 can be rotatably driven upwards in rotational velocity br in a backwardly downwardly bd direction by the user &# 39 ; s stepping on a stair surface ss or by exerting the force of the user &# 39 ; s weight wt or a downwardly user exerted force df , fig1 by the user &# 39 ; s muscles . the apparatus 5 is adapted so that the treadmill 20 can be driven from 0 rotational speed upwardly to a preselected rotational speed br at which preselected rotational speed a resistance device 100 begins to exert a resistance to rotation br . the treadmill 20 is preferably interconnected to a resistance device 100 , such as an eddy current brake device that exerts a resistance to rotation br that increases exponentially or non - linearly with an increase of the rotational speed br of the treadmill or belt 20 . as shown in fig1 - 4 , the treadmill 20 is comprised of a plurality of serially or successively pivotably interconnected stairs 70 , each stair comprising a forward link 50 and rear link 60 that are pivotably or hingedly interconnected along a line 72 that extends across the width w of the step surface ss . the hinge along line 72 is formed by a series of complementary forward fingers 72 ff ( each comprised of lateral fingers 300 f , fig1 a , 12b ) and forward recesses 72 fr formed on rear edge of the forward link 50 that mate with a complementary series of rearward recesses 72 rr and rearward fingers 72 rf ( each comprised of lateral fingers 302 f ) formed on a forward edge of the rear link 60 . a connecting pin 72 p is insertable through a complementary aperture pa provided in fingers 72 ff and 72 rf so as to pivotably connect the forward link 50 to the rear link 60 along the hinge line 72 . the step hinge line 72 is disposed some selected distance along the overall step length sl that extends between the forward edge 74 f and the rear edge 74 r of each step surface ss of each stair 70 . the step surface ss is comprised of a forward step surface ssf formed on the upper surface of the forward link 50 and a rearward step surface ssr formed on the upper surface of the rear link 60 . the forward ssf and rear ssr step surfaces are generally planar such that the two surfaces ssf and ssr form an overall step surface ss that is generally continuous and planar when the forward 50 and rear 60 links pivot into angular alignment with each other along a portion pot of the overall path of rotational travel of the treadmill 20 . preferably , the hinge line 72 is spaced a distance rsl of between about 1 and about 5 inches from the rear edge 72 of a step surface ss . typically , the overall length sl of a step surface is between about 10 and about 15 inches . each stair 70 is also comprised of an upper riser ur and a lower riser lr . the upper riser ur of each stair 70 extends downwardly from the rear edge 74 r of the stair surface ss and terminates at a lower edge in a second hinge line 82 where the ur riser portion of the stair 70 pivotably interconnects to an upper edge of the lower riser lr portion of stair 70 . the lower riser portion lr of stair 70 comprises an upward extension of the first link 50 s of an immediately successive stair 70 s to the immediately preceding stair 70 as shown in fig2 . the lower riser lr extends upwardly from the forward step surface ssf such that the riser hinge line 82 of the lower riser is disposed a normal height or distance of between about 1 and about 5 inches above or away from the forward step surface ssf . similar to hinge line 72 , the hinge is formed along riser hinge line 82 by the mating of complementary upper recesses 82 uf formed on the lower edge of link 60 with lower riser fingers 82 lf formed on the upper edge of the lower riser lr . a connecting pin 82 p is insertable through a complementary aperture pa provided in fingers 82 uf and 82 lf so as to pivotably connect the forward link 50 s to the rear link 60 along the hinge line 82 . fig5 is a side view of the fig4 components showing the forward and rear links disengaged from each other and showing details of the hinge components . the forward link 50 includes a series of forward fingers 72 ff each having a curved connection portion 90 f ( shown in dashed lines ). forward link 50 also includes a series of forward recesses 72 fr each having at least one curved guide unit 94 f and a concave surface 96 f ( both shown in dashed lines ). similarly , the rear link 60 includes a series of rearward fingers 72 rf each having a curved connection portion 90 r . furthermore , rear link 60 includes a series of rearward recesses 72 rr each having at least one curved guide unit 94 r and a concave surface 96 r . the curved connection portions 90 f and 90 r angularly extend from the generally planar step surface toward the lower surface of the step . in some embodiments , the curved connection portions 90 f and 90 r can extend from the generally planar step surface along an arc between about 45 degrees and about 135 degrees . as shown in fig6 - 8 , each of the curved connection portions is rotatably received between the complementary curved guide unit ( s ) and concave surface to pivotably interconnect the forward link 50 to the rear link 60 via forward and rearward fingers 72 ff and 72 rf . a connecting pin 72 p is insertable through a complementary aperture pa provided in fingers 72 ff and 72 rf to secure the connection between the forward link 50 and the rear link 60 along the hinge line 72 . once connected , the forward and rearward fingers 72 ff and 72 rf completely enclose their complementary forward and rearward recesses to form a continuous exterior surface throughout all angles of rotation of the pivotable interconnection . when the forward and rearward step surfaces are substantially perpendicular to one another , they can be connected by their hinge as shown in fig6 . each curved connection portion 90 f included in the forward fingers 72 ff of the forward link 50 is rotatably received between the complementary curved guide unit ( s ) 94 r and concave surface 96 r included in the complementary rearward recess 72 rr of the rear link 60 . similarly , each curved connection portion 90 r in the rearward fingers 72 rf disposed on the rear link 60 is rotatably received between the complementary curved guide unit ( s ) 94 f and concave surface 96 f in the complementary forward recess 72 fr disposed on the forward link 50 . furthermore , fig6 shows that each of the forward and rearward fingers 72 ff and 72 rf completely encloses their complementary forward and rearward recesses 72 fr and 72 rr to form a continuous exterior surface 104 . fig7 is a side view showing the front and rear links in an angular position where the step surfaces are angled at about 45 degrees relative to each other . in comparison with fig6 , it can be seen that each of the curved connection portions 90 f and 90 r is further rotatably engaged between the complementary curved guide unit ( s ) and concave surface . moreover , the forward and rearward recesses 72 fr and 72 rr remain completely enclosed by each of the forward and rearward fingers 72 ff and 72 rf thereby maintaining the continuous exterior surface 104 . fig8 is a side view similar to fig6 and 7 showing the front and rear links in an angular position where the step surfaces are coplanar . in this configuration , each of the curved connection portions 90 f and 90 r is fully rotatably engaged between the complementary curved guide unit and concave surface . when the front and rear step components are coplanar , a radially inward protrusion in each of the curved connection portions 90 f and 90 r contacts a front face of the complementary curved guide unit to form an abutment 108 . furthermore , fig8 shows that each of the forward and rearward fingers 72 ff and 72 rf still completely encloses their complementary forward and rearward recesses 72 fr and 72 rr and the continuous exterior surface 104 remains . fig9 and 11 show a comparison between stair 70 having a hinge disposed along hinge line 72 and a stair 120 ( shown in dashed lines ) having a hinge disposed along hinge line 112 . hinge line 72 is disposed some selected distance along the overall step length that extends between the forward edge 74 f and the rear edge 74 r of the step surface of stair 70 . on the other hand , hinge line 112 is disposed on the rear edge 74 r of the step surface . each of stairs 70 and 120 are pivotably connected to a belt or chain 40 at points 122 and 124 and are configured to move along a path outlined by belt or chain 40 in a backward and downward direction bd . fig9 illustrates a first stair position in which both stair 70 and stair 120 form a generally continuous and planar step surface . as a user steps on the step surface of stairs 70 and 120 , the user &# 39 ; s weight or a downwardly user exerted force causes each of stairs 70 and 120 to move in a backward and downward direction bd into a second stair position illustrated in fig1 . in the second stair position , stairs 70 and 120 have pivoted about their respective hinge lines 72 and 112 to accommodate the path outlined by the chain or belt 40 . due to the difference in the position of hinge lines 72 and 112 , stair 120 extends a distance h below stair 70 . fig1 shows in schematic a single stair 70 of the apparatus of fig1 - 10 mounted on a chain 40 that is driven by the pulley 30 r and in particular the path of travel epn of the outermost edge 74 r of a stair 70 according the fig1 - 10 apparatus relative to the path of travel epc of the outermost edge 700 e of a conventional treadmill stair 500 , 600 . as shown , the radial distance rdn to which the outermost edge 74 r of a stair 70 according to the invention extends from the center of the pulley 38 r is significantly reduced or less relative to the radial distance rdc to which the outermost edge of a conventional stair 500 , 600 extends where the conventional stair 500 , 600 has about the same step length csl and riser length crl as the step length ( fsl plus rsl ) and riser length ( url plus lrl ) of a stair 70 according to the invention . such a reduction in distance of extension rdn relative to conventional stair distance of extension rdc . such reduction in extension rdn in particular reduces the vertical height h of the outermost edge 74 r of the stair 70 off of the ground g when a stair 70 is in its closest to ground g position at the most vertical v disposition over the course of rotation of the treadmill 20 around its angled path of travel . such reduction and thus reduces the minimum height off of the ground by which the stair surface ss can be mounted which in turn reduces the overall height of the apparatus relative to ground g and the height off the ground that a user must step up in order to step on the lowest stair on the treadmill 20 when the apparatus 5 is mounted on the frame 10 . fig1 a , 12b show the details of a preferred hinge 72 structure of an apparatus 5 according to the invention . as shown the hinge 72 comprises at least one male hinge member 300 disposed on one of the rear edge 50 e of the step and the front edge 60 e of the riser and at least a pair of male hinge members 302 disposed on the other of the rear edge 50 e of the step and the front edge 60 e of the riser , the pair of male hinge members 302 being spaced apart to form an aperture or recess 7 rr that is complementary to and receives the at least one male hinge member 300 . each of the male hinge members 300 , 302 comprises a center arcuate member 300 c , 302 c having an outside wall 300 w , 302 w that extends continuously from the upper step surface ss of the step and the outside surface os of the riser at least about 180 degrees ( angle b ) around the rear 50 e and front 60 e edges of the step 50 and the riser 60 respectively . each of the male hinge members 300 , 302 further comprises a pair of lateral arcuate fingers 300 f , 302 f disposed on opposing lateral sides of the center arcuate member 300 c , 302 c , the lateral arcuate fingers 300 f , 302 f having an outside surface 300 fw , 302 fw that extends continuously from the upper step surface ss of the step 50 and from the outside surface os of the riser about 90 degrees ( angle c ) around the rear 50 e and front edges 60 e of the step and the riser . the male hinge members 300 , 302 each have a longitudinal aperture 300 ap , 302 ap with a common axis apx for receiving a pin or rod 72 p that extends through the longitudinal aperture 300 ap , 302 ap along the axis apx and pivotably interconnects the step 50 to the riser 60 forming the hinge 72 . the longitudinal apertures 300 ap , 302 ap , the center arcuate members 300 c , 302 c and the lateral fingers 300 f , 302 f are adapted to arrange the outside wall 300 w , 302 w of the center arcuate member 300 c , 302 c and the outside surfaces 300 fw , 302 fw of the lateral arcuate fingers 300 f , 302 f such that a continuous surface is formed around the hinge when the step 50 and the riser 60 are pivoted relative to each other around at least 90 degrees .

a stair stepping apparatus comprising : a plurality of stairs forming a treadmill or endless belt on which a user steps , wherein each stair comprises : a first link comprised of a major step portion joined to a minor riser portion by a rigid elbow , and a second link comprised of major riser portion joined to a minor step portion by a rigid elbow .

the methods and compositions of the invention are useful for increasing the sensitivity of cells to chemotherapeutic agents such as , for example , anti - cancer agents like paclitaxel , analgesics , opiates , hormones or antibiotics and the like . in particular the methods and compositions of the invention are useful for increasing the sensitivity of cells associated with cellular proliferative disorders ( eg ., a neoplasm ) by increasing the efficacy without concomitant toxicity to non - cancer cells the invention provides methods and compositions useful for treating tumors and preventing or reducing the chances of relapse and death as a result of cytotoxicity . in addition , the invention eliminates or reduces the number of multidrug resistant cells by eliminating cancer cells prior to any mutation inducing a multidrug resistant phenotype . accordingly , by reducing multi - drug resistant tumor cells from arising , the invention satisfies the shortcomings of current therapeutic modalities . the term “ subject ” as used herein refers to any animal having a disease or condition which requires treatment with a chemotherapeutic agent wherein the chemotherapeutic agent has reduced efficacy relative to that desired . preferably the subject is suffering from a cellular proliferative disorder ( eg ., a neoplastic disorder ). subjects for the purposes of the invention include , but are not limited to , mammals ( eg ., bovine , canine , equine , feline , porcine ) and preferably humans . by “ cell proliferative disorder ” is meant that a cell or cells demonstrate abnormal growth , typically aberrant growth , leading to a neoplasm , tumor or a cancer . cell proliferative disorders include , for example , cancers of the breast , lung , prostate , kidney , skin , neural , ovary , uterus , liver , pancreas , epithelial , gastric , intestinal , exocrine , endocrine , lymphatic , haematopoietic system or head and neck tissue . generally , neoplastic diseases are conditions in which abnormal proliferation of cells results in a mass of tissue called a neoplasm or tumor . neoplasms have varying degrees of abnormalities in structure and behaviour . some neoplasms are benign while others are malignant or cancerous . an effective treatment of neoplastic disease would be considered a valuable contribution to the search for cancer preventive or curative procedures . the methods of this invention involve in one embodiment , ( 1 ) the administration of hyaluronan , prior to , together with , or subsequent to the administration of a chemotherapeutic agent ; or ( 2 ) the administration of a combination of hyaluronan and a chemotherapeutic agent . as used herein , the term “ therapeutically effective amount ” is meant an amount of a compound of the present invention effective to yield a desired therapeutic response . for example to prevent cancer or treat the symptoms of cancer in a host or an amount effective to treat cancer . the specific “ therapeutically effective amount ” will , obviously , vary with such factors as the particular condition being treated , the physical condition of the patient , the type of mammal being treated , the duration of the treatment , the nature of concurrent therapy ( if any ), and the specific formulations employed and the structure of the compounds or its derivatives . as used herein , a “ pharmaceutical carrier ” is a pharmaceutically acceptable solvent , suspending agent or vehicle for delivering the hyaluronan and / or chemotherapeutic agent to the animal or human . the carrier may be liquid or solid and is selected with the planned manner of administration in mind . as used herein , “ cancer ” refers to all types of cancers or neoplasm or malignant tumours found in mammals . cancer includes sarcomas , lymphomas and other cancers . the following types are examples , but are , but is not intended to be limited to these particular types of cancers : prostate , colon , breast , both the mx - 1 and the mcf lines , pancreatic , neuroblastoma , rhabdomysarcoma , home , lung , murine , melanoma , leukemia , pancreatic , melanoma , ovarian , brain , head & amp ; neck , kidney , mesothelioma , sarcoma , kaposi &# 39 ; s , sarcoma , stomach , and uterine . as used herein , the term “ cell ” include but is not limited to mammalian cells ( eg ., mouse cells rat cells or human cells ). the hyaluronan and / or chemotherapeutic agents may be administered orally , topically , or parenterally in dosage unit formulations containing conventional non - toxic pharmaceutically acceptable carriers , adjuvants , and vehicles . the term parenteral as used herein includes subcutaneous injections , aerosol , intravenous , intramuscular , intrathecal , intracranial , intrasternal injection or infusion techniques . the present invention also provides suitable topical , oral , and parenteral pharmaceutical formulations for use in the novel methods of treatment of the present invention . the compounds of the present invention may be administered orally as tablets , aqueous or oily suspensions , lozenges , troches , powders , granules , emulsions , capsules , syrups or elixirs . the composition for oral use may contain one or more agents selected from the group of sweetening agents , flavouring agents , colouring agents and preserving agents in order to produce pharmaceutically elegant and palatable preparations . the tablets contain the active ingredient in admixture with non - toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets . these excipients may be , for example , ( 1 ) inert diluents , such as calcium carbonate , lactose , calcium phosphate or sodium phosphate ; ( 2 ) granulating and disintegrating agents , such as corn starch or alginic acid ; ( 3 ) binding agents , such as starch , gelatin or acacia ; and ( 4 ) lubricating agents , such as magnesium stearate , stearic acid or talc . these tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period . for example , a time delay material such as glyceryl monostearate or glyceryl distearate may be employed . coating may also be performed using techniques described in the u . s . pat . nos . 4 , 256 , 108 ; 4 , 160 , 452 ; and 4 , 265 , 874 to form osmotic therapeutic tablets for control release . the hyaluronan as well as the chemotherapeutic agents useful in the method of the invention can be administered , for in vivo application , parenterally by injection or by gradual perfusion over time independently or together . administration may be intravenously , intraperitoneally , intramuscularly , subcutaneously , intracavity , or transdermally . for in vitro studies the agents may be added or dissolved in an appropriate biologically acceptable buffer and added to a cell or tissue . preparations for parenteral administration include sterile aqueous or non - aqueous solutions , suspensions , and emulsions . examples of non - aqueous solvents are propylene glycol , polyethylene glycol , vegetable oils such as olive oil , and injectable organic esters such as ethyl oleate . aqueous carriers include water , alcoholic / aqueous solutions , emulsions or suspensions , including saline and buffered media . parenteral vehicles include sodium chloride solution , ringer &# 39 ; s dextrose , dextrose and sodium chloride , lactated ringer &# 39 ; s intravenous vehicles include fluid and nutrient replenishers , electrolyte replenishers ( such as those based on ringer &# 39 ; s dextrose ), and the like . preservatives and other additives may also be present such as , for example , anti - microbials , anti - oxidants , chelating agents , growth factors and inert gases and the like . it is envisioned that the invention can be used to treat pathologies associated cell proliferative disorders , including , for example , neoplasms , cancers ( eg ., cancers of the breast , lung , prostate , kidney , skin , neural , ovary , uterus , liver , pancreas , epithelial , gastric , intestinal , exocrine , endocrine , lymphatic , haematopoietic system or head and neck tissue ), fibrotic disorders and the like . the methods and compounds of the invention may also be used to treat other diseases associated with chemotherapeutic treatment such as neurodegenerative disorders , hormonal imbalance and the like . therefore , the present invention encompasses methods for ameliorating a disorder associated with cell proliferation , neoplasms , cancers and the like , including treating a subject having the disorder , at the site of the disorder , with hyaluronan and a chemotherapeutic agent in an amount sufficient to inhibit or ameliorate the cell &# 39 ; s proliferation or the disorder . generally , the terms “ treating ”, “ treatment ” and the like are used herein to mean affecting a subject , tissue or cell to obtain a desired pharmacologic and / or physiologic effect . the effect may be prophylactic in terms of completely or partially preventing a cell proliferative disorder or sign or symptom thereof , and / or may be therapeutic in terms of a partial or complete cure for a disorder and / or adverse effect attributable to , for example , aberrant cell proliferation . “ treating ” as used herein covers any treatment of , or prevention of a cell proliferative disorder in a vertebrate , a mammal , particularly a human , and includes : ( a ) preventing the disorder from occurring in a subject that may be predisposed to the disorder , but has not yet been diagnosed as having it ; ( b ) inhibiting the disorder , i . e ., arresting its development ; or ( c ) relieving or ameliorating the disorder , i . e ., cause regression of the disorder . the invention includes various pharmaceutical compositions useful for ameliorating cell proliferative disorder , including neoplasms , cancers and the like . the pharmaceutical compositions according to one embodiment of the invention are prepared by bringing hyaluronan , analogue , derivatives or salts thereof and one or more chemotherapeutic agents or combinations of hyaluronan and one or more chemotherapeutic agents into a form suitable for administration to a subject using carriers , excipients and additives or auxiliaries . frequently used carriers or auxiliaries include magnesium carbonate , titanium dioxide , lactose , mannitol and other sugars , talc , milk protein , gelatin , starch , vitamins , cellulose and its derivatives , animal and vegetable oils , polyethylene glycols and solvents , such as sterile water , alcohols , glycerol and polyhydric alcohols . intravenous vehicles include fluid and nutrient replenishers . preservatives include antimicrobial , anti - oxidants , chelating agents and inert gases . other pharmaceutically acceptable carriers include aqueous solutions , non - toxic excipients , including salts , preservatives , buffers and the like , as described , for instance , in remington &# 39 ; s pharmaceutical sciences , 15th ed . easton : mack publishing co ., 1405 - 1412 , 1461 - 1487 ( 1975 ) and the national formulary xiv ., 14th ed . washington : american pharmaceutical association ( 1975 ), the contents of which are hereby incorporated by reference . the ph and exact concentration of the various components of the pharmaceutical composition are adjusted according to routine skills in the art . see goodman and gilman &# 39 ; s the pharmacological basis for therapeutics ( 7th ed .). the pharmaceutical compositions are preferably prepared and administered in dose units . solid dose units are tablets , capsules and suppositories . for treatment of a subject , depending on activity of the compound , manner of administration , nature and severity of the disorder , age and body weight of the subject , different daily doses can be used . under certain circumstances , however , higher or lower daily doses may be appropriate . the administration of the daily dose can be carried out both by single administration in the form of an individual dose unit or else several smaller dose units and also by multiple administration of subdivided doses at specific intervals . the pharmaceutical compositions according to the invention may be administered locally or systemically in a therapeutically effective dose . amounts effective for this use will , of course , depend on the severity of the disease and the weight and general state of the subject . typically , dosages used in vitro may provide useful guidance in the amounts useful for in situ administration of the pharmaceutical composition , and animal models may be used to determine effective dosages for treatment of particular disorders . various considerations are described , eg ., in langer , science , 249 : 1527 , ( 1990 ). formulations for oral use may be in the form of hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent , for example , calcium carbonate , calcium phosphate or kaolin . they may also be in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium , such as peanut oil , liquid paraffin or olive oil . aqueous suspensions normally contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspension . such excipients may be ( 1 ) suspending agent such as sodium carboxymethyl cellulose , methyl cellulose , hydroxypropylmethylcellulose , sodium alginate , polyvinylpyrrolidone , gum tragacanth and gum acacia ; ( 2 ) dispersing or wetting agents which may be ( a ) naturally occurring phosphatide such as lecithin ; ( b ) a condensation product of an alkylene oxide with a fatty acid , for example , polyoxyethylene stearate ; ( c ) a condensation product of ethylene oxide with a long chain aliphatic alcohol , for example , heptadecaethylenoxycetanol ; ( d ) a condensation product of ethylene oxide with a partial ester derived from a fatty acid and hexitol such as polyoxyethylene sorbitol monooleate , or ( e ) a condensation product of ethylene oxide with a partial ester derived from fatty acids and hexitol anhydrides , for example polyoxyethylene sorbitan monooleate . the pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension . this suspension may be formulated according to known methods using those suitable dispersing or wetting agents and suspending agents which have been mentioned above . the sterile injectable preparation may also a sterile injectable solution or suspension in a non - toxic parenterally - acceptable diluent or solvent , for example , as a solution in 1 , 3 - butanediol . among the acceptable vehicles and solvents that may be employed are water , ringer &# 39 ; s solution , and isotonic sodium chloride solution . in addition , sterile , fixed oils are conventionally employed as a solvent or suspending medium . for this purpose , any bland fixed oil may be employed including synthetic mono - or diglycerides . in addition , fatty acids such as oleic acid find use in the preparation of injectables . hyaluronan together with a chemotherapeutic agent of the present invention may also be administered in the form of liposome delivery systems , such as small unilamellar vesicles , large unilamellar vesicles , and multilamellar vesicles . liposomes can be formed from a variety of phospholipids , such as cholesterol , stearylamine , or phosphatidylcholines . dosage levels of the compounds of the present invention are of the order of about 0 . 5 mg to about 10 mg per kilogram body weight , with a preferred dosage range between about 5 mg to about 20 mg per kilogram body weight per day ( from about 0 . 3 gms to about 1 . 2 gms per patient per day ). the amount of active ingredient that may be combined with the carrier materials to produce a single dosage will vary depending upon the host treated and the particular mode of administration . for example , a formulation intended for oral administration to humans may contain about 5 mg to 1 g of an active compound with an appropriate and convenient amount of carrier material which may vary from about 5 to 95 percent of the total composition . dosage unit forms will generally contain between from about 5 mg to 500 mg of active ingredient . it will be understood , however , that the specific dose level for any particular patient will depend upon a variety of factors including the activity of the specific compound employed , the age , body weight , general health , sex , diet , time of administration , route of administration , rate of excretion , drug combination and the severity of the particular disease undergoing therapy . in addition , some of the compounds of the instant invention may form solvates with water or common organic solvents . such solvates are encompassed within the scope of the invention . the compounds of the present invention may additionally be combined with other compounds to provide an operative combination . it is intended to include any chemically compatible combination of chemotherapeutic agents , as long as the combination does not eliminate the activity of the hyaluronan of this invention . the invention will now be further described by way of reference only to the following non - limiting examples . it should be understood , however , that the examples following are illustrative only , and should not be taken in any way as a restriction on the generality of the invention described above . in particular , while the invention is described in detail in relation to cancer , it will be clearly understood that the findings herein are not limited to treatment of cancer . for example , ha may be used for treatment of other conditions . ha used in all of the in vitro and in vivo studies were obtained from kyowa hakko kogyo ( yamaguchi , japan ). 5 - fu was obtained from sigma , st . louis , usa . and adraimycin from cytomix , northcote , melbourne , australia . a standard profile of the ha used is shown in table 1 . a 10 mg / ml stock of ha solution was prepared by dissolving desiccated ha ( modal mr 7 . 5 × 10 5 kda ) in pyrogen - free injection grade water . to ensure a homogenous solution the ha was dissolved overnight at 4 ° c . followed by thorough vortexing . to ensure that the ha had maintained its molecular weight during the preparation of the stock solution , the solution was analysed on a sephacryl s - 1000 size exclusion gel with column specifications of 1 . 6 cm × 70 cm , sample size 2 ml , flow rate 18 ml / h and 2 ml fraction size . hyaluronan was detected in column fractions by the uronic acid assay . the uronic acid assay was used to detect the presence of hyaluronan qualitatively from the fractions collected from the gel filtration chromatography procedure . a 25 μl aliquot of each fraction was then transferred into a 96 well plate . 250 μl of a carbazole reagent ( 3m carbazole / 0 . 025m borate in h 2 so 4 ) was then added to these fractions . the 96 well plate was incubated for 45 - 60 min at 80 ° c . a dynatech mr7000 plate reader with a 550 nm filter was used to read the 96 well plate . the absorbance was considered to be significant when it was & gt ; 3 standard deviations above the background absorbance . the background was calculated by taking an equal number of sample points before and after v o and v t where the average number taken was 16 ( fraser et al . 1998 ). a stock solution of 5 - fu was prepared by dissolving powdered 5 - fu in 01m naoh ( ph 8 . 9 ) and brought to a concentration of 1 mg / ml with pyrogen - free injection grade 0 . 9 % w / v nacl . the stock solution was filtered through a 0 . 22 μm filter to ensure sterility . the 5 - fu was diluted by adding the required volume of stock solution to the cell - line specific growth medium as specified above . a 10 mg / ml solution of adriamycin in 0 . 9 % nacl was obtained from cytomix . human breast adenocarcinoma cell lines mda - mb - 468 , mda - mb - 435 and mda - mb - 231 were selected based on ha binding affinity ( culty et al , 1994 ), and the expression of the ha receptors of cd44 and rhamm ( wang et al , 1996 ). the characteristics of these cell lines are shown in table 2 . cell lines mda - mb - 468 , mda - mb - 435 and mda - mb - 231 were routinely grown and subcultured as a monolayer in 175 cm 2 culture flasks in leibovitz l - 15 medium supplemented with 10 % foetal calf serum ( fcs ) and antibiotic / antimycotic reagents at 37 ° c . in humidity controlled incubator with 100 % ( v / v ) air . leibovitz - l - 15 with glutamine ( 10 × concentrate ) rpmi ( 10 × concentrate ), eagles basal medium ( ebm , 10 × concentrate ), 20 mm hepes , 0 . 09 % w / v bicarbonate , hanks &# 39 ; balanced salt solution ( hbss , 10 × concentrate ) and dulbecco &# 39 ; s phosphate buffered saline without calcium and magnesium ( pbs , 10 × concentrate ) were purchased from sigma ( st louis , mo ., usa ). powder concentrates were dissolved in the required volume of reverse osmosis deionised pyrogen - free distilled water to make a single strength solution , sterilised by 0 . 22 μm high pressure filtration ( millipore corporation , ma . u . s . a . ), and stored at 4 ° c . fcs ) were purchased from the csl ltd ., australia . fcs was stored at − 20 ° c . antibiotic / antimycotic solution ( 100 × concentrate ) containing 10 , 000 units penicillin , 10 mg streptomycin and 25 μg amphotericin u / ml was obtained from sigma ( st louis , usa ). trypsin / edta solution ( 10 × concentrate ) containing 5 g porcine trypsin and 2 g edta / l in 0 . 9 % w / v sodium chloride was obtained from sigma ( st louis , mo ., usa ). all breast cancer cell lines were purchased from american tissue culture collection ( rockville , usa ). all plastic disposable culture vessels were purchased from greiner ( austria ). eight - welled , tissue culture microscope slides were obtained from linbro ( flow laboratories , va , usa ). for the tests , mda mb - 468 , mda mb - 231 and mda mb - 435 cell line were grown in 90 % leibovitz l - 15 medium supplemented with 10 % fcs . when confluent the cultures were washed 1 × in hbss and trypsinised in 0 . 25 % trypsin / 0 . 05 % edta . the cell suspensions were counted with an automated cell counter ( zm - 2 coulter counter ) by adding 15 ml saline + 0 . 2 ml of cell suspension . the cells were plated into 48 - well plates ( 1 cm 2 surface area ) by adding 1 ml of cell suspension per well . cells were allowed to attach for 24 h , before the media was removed , monolayers washed . the test media was ; growth media containing 0 - 1 μm adriamycin or 5 - fluorouracil with or without the addition of 0 - 1 μm of ha ( modal mw 750 , 000 ). the cells were exposed to the several combinations of ha and drugs for different times and at different concentrations ( table 3 ). after the incubation and growth periods the cell monolayers were washed with hbss and trypsinised in 0 . 25 % trypsin / 0 . 05 % edta . the cell suspensions were counted with an automated cell counter ( zm - 2 coulter counter ) by adding 15 ml saline + 0 . 2 ml of cell suspension . results were expressed as % of no drug control which was calculated as : cell ⁢ ⁢ count × 100 cells ⁢ ⁢ in ⁢ ⁢ no ⁢ ⁢ drug ⁢ ⁢ control or depending on the experiment as % of drug control , calculated as : exponentially growing human breast cancer cells mda mb 231 as described in example 2 were incubated with 0 - 5 mg / ml ha ( modal mr 750 , 000 d ) for 24 h . at 24 h the cells were counted and photographed with cpr , 1600 film rolls from eastman kodak company , rochester , usa . when ha was incubated with breast cancer cells for 30 min , 1 h , 24 h or 3 days a varied response was observed , where the reduction in breast cancer cell number ranged from 0 - 29 % ( see table 4 ). when human breast cancer cells were incubated with ha specific morphological changes ( see fig1 ) were also observed such as swelling of the plasma membrane , greater granularity of cytosolic components . when human breast cancer cells were exposed to ha for 30 min , 1 h , 24 h or 3 days followed by exposure toadriamycin , it became evident that ha ehanced the cytotoxicity of the drug ( fig3 & amp ; table 5 ). all figures represent the range of 2 - 3 separate experiments , where the numerical values are the multiplication factor decrease in ic 50 which is exerted by the addition of ha to drug or pre - sensitization of cancer cells with ha before the addition of drug . based on the results from the in vitro drug sensitivity experiments in example 2 , evaluation of the treatment efficacy of hyaluronan as a sole agent , and as a chemosensitizer in the treatment human breast carcinomas in vivo was undertaken . from the results in example 2 the carcinoma cell line mda - mb - 468 was selected as the cancer cell inoculant for the generation of any nude mouse human tumour xenografts . cells were routinely grown and subcultured as a previously described in example 2 . for injection into mice , cells were grown to 100 % confluency , trypsinised in 0 . 025 % trypsin / 0 . 01 % edta solution , washed twice by centrifugation in a beckman tj - 6 bench centrifuge at 400 g av for 10 min , counted using a model - zm coulter counter and resuspended in serum - free leibovitz l - 15 medium at 1 × 10 8 cells / ml . six to eight weeks old athymic cba / wehi nude female mice , purchased from the walter and eliza hall research institute , melbourne australia , were maintained under specific pathogen - free conditions , with sterilised food and water available ad libitum . each mouse received one injection containing 5 × 10 6 cells in 50 μl . the cells were injected with a 26 gauge needle into the mammary fat pad directly under the first nipple ( lamszus et al , 1997 ). tumour measurements were made weekly by measuring three perpendicular diameters ( d 1 d 2 d 3 ). tumour volume was estimated using the formula : treatment with 5 - fu ± ha was commenced approximately 4 - 8 weeks after the cancer cell inoculation . the mean tumour size for mice used in each study is summarised in table 6 . approximately 8 weeks after tumour induction two tumour - bearing mice were given a lethal dose of nembutal . within 3 min of killing the mice , tumours were surgically removed and immediately fixed in 10 % buffered formalin for 12 h . the fixed tumour was dehydrated overnight in a series of 70 - 100 % ethanol , followed by paraffin embedding from which 2 - 4 μm sections were cut . the sections were placed on slides , de - waxed , and brought to water . slides were washed 3 × 5 min in pbs . heterophile proteins were blocked by incubation with 10 % foetal calf serum for 10 min , followed by a pbs rinse . secondary antibodies used in the visualisation of ha and ha synthase antibodies were purchased from dako ( california , u . s . a .). 3 , 3 ′- diaminobenzidine ( sigma fast dab ) tablets were obtained from sigma , st . louis , usa . the detection antibodies were applied for 60 min at rt . the detection antisera or antibodies were against rhamm , cd44h and cae . the slides were washed 3 × 5 min in pbs and endogenous peroxidase activity blocked by immersion in 0 . 3 % h 2 o 2 in methanol for 20 min . following a further pbs wash , the peroxidase - conjugated swine anti - rabbit secondary antiserum was applied for 60 min at rt , followed by 3 × 5 min washes in pbs . sigma fast 3 , 3 ′- diaminobenzidine tablets ( dab ) were prepared according to the manufacturer &# 39 ; s instructions and the dab solution was applied for 5 - 10 min at rt . the slides were washed in tap water for 10 min , counterstained with haematoxylin , dehydrated and mounted . individual injections of 5 - fu were prepared according to individual mouse masses , with the aim of delivering 30 mg / kg 5 - fu in 50 μl ( equivalent to human therapeutic dose of 10 . 5 mg / kg for a mean body weight of 60 kg ; inaba et al , 1988 ). ha injection comprising a final ha concentration equivalent to 12 . 5 mg / kg of mouse mass were prepared so that deliver of 12 . 5 mg / kg ha in 50 μl could be effected . with this quantity of ha injected into the body , saturation kinetics would be observed for the period of the experimentation ( fraser et al , 1983 ). one of the most commonly used treatment regimens for human breast cancer is cyclophosphamide , methotrexate and 5 - fluorouacil , which is administered on day 1 and 8 of a 28 day cycle in human breast cancer the initial treatment regimen is for 6 cycles at which time the patient condition is re - assessed , therefore we tried to simulate the human treatment regimen as closely as possible by exposing the mice to 6 cycles ( 6 months ) of treatment in a long term efficacy study and a 6 cycles ( 6 week ) short term efficacy study . considering the life cycle of a mouse is approximately 2 years we commenced both short - term and long - term treatment protocols ( see table 7 ). mice were randomly divided into 7 groups of 8 animals per group for the short term study and 5 groups of 8 animals for the long term study ( refer to table 7 for dosage and treatment administration schedule ). the treatment was not extended over the 6 month regimen since it has been demonstrated that chemotherapy lasting more than six months has not generally been associated with greater benefit ( harris et al , 1992 ). animals were weighed and tumour volumes measured on the day of treatment application for long term study . in the 6 - week study animals were weighed and tumour volumes measured on a daily basis . animals were individually placed in an injection box , and the injections were administered via the tail vein . it has been experimentally proven that stress can be a major factor in a patients response to chemotherapy ( shackney et al , 1978 ), therefore we ensured that equal numbers of mice were allocated to each cage , the animal number per cage varied from 5 - 8 depending on the stage of experimentation . the experimental end - point occurred when the animal had to be euthanised due to degree of disease progression or when the 6 month ( long term ) or 6 week ( short term ) treatment regimen was completed . due to the animal ethics guidelines the animals were monitored fortnightly by an independent animal ethics officer who assessed the degree of disease progression . the following criteria were used to determine if an animal had reached the stage of experimental end - point of necessary death : 2 ). animal was not eating or drinking and had experienced dramatic weight loss ; or at the experimental end - point the animals were anaesthetized by a 0 . 1 ml intra - peritoneal injection of nembutal ( 60 mg / ml ), blood was collected followed by killing of the animals using cervical dislocation . immediately after killing the mouse the tumour , liver , heart , spleen , bladder , left and right kidneys , uterus , lungs , stomach , intestines , brain and lymph nodes were excised and placed in 4 % formalin buffered with 0 . 06m phosphate ph 7 . 5 , and cetylpyridinium chloride , 1 . 0 % w / v . the tissue was fixed for 16 - 24 h before histological processing . fixed tissue was dehydrated stepwise to 100 % ethanol and embedded in paraffin blocks from which 2 - 4 μm sections were placed on glass microscope slides . staining the tissue sections with a haematoxylin nuclear stain and eosin cytoplasmic stain highlighted any pathological features that could indicate treatment toxicity . nine to 11 lymph nodes were collected per animal , ensuring that all nodes which drained the tumour area were collected . there are currently two methods used for the detection of lymph node metastasis both methods of metastasis detection were employed in this study . not all commercially available cea antibodies react with human breast cancer cells , so we tested the reactivity of 5 different antibodies ( dako , amersham and kpl ). the haematoxylin and eosin stained lymph nodes were examined by dr p . allen ( certified pathologist ) where each node was microscopically examined for the presence of tumour cells . the cea immunostained lymph nodes were microscopically examined , where any positively stained nodes were counted and considered positive for lymph node metastasis . tumour volume was monitored on a daily or weekly basis by calliper measurements and tumour volume calculated as previously described . at the end of the 6 week study , tumour mass was determined where the ha chemosensitizing therapy had significantly smaller tumours than the saline group , ha and 5 - fu groups ( p = 0 . 005 ) as seen in fig6 . no significant differences in tumour response were noted in the initial 2 weeks of treatment , but thereafter the ha followed by 5 - fu tumour growth was retarded in comparison to the other treatment groups . during the 6 weeks of treatment interesting differences were noted in the number of tumour doubling cycles . mice receiving the saline treatment underwent an average of 4 tumour doublings , while the incorporation of ha into the treatment regimen significantly increased the tumour doubling time where ha / 5 - fu animals underwent an average of one tumour doubling cycle , once again highlighting the effect of ha on 5 - fu cytotoxicity . all animals displayed lymph node metastasis in lymph nodes that were adjacent to the primary tumour . the percentage of lymph node involvement ( number of metastatic nodes per animal ) was greatly reduced by the ha followed by 5 - fu , 5 - fu and ha treatment , where the saline group demonstrated a 6 - fold increase in the amount of lymph node involvement . the other treatment groups demonstrated a significantly smaller percentage at 12 . 2 - 14 . 3 % ( dunnett &# 39 ; s multiple comparison test , p =& lt ; 0 . 001 ). the co - administration of ha resulted in a significant reduction in non - lymphoid metastasis . with the exception of the mice receiving the ha therapy , new tumours were observed around the neck or underarm region of the area adjacent to the primary tumour . one of the most common toxic effects of 5 - fu is on the gastro - intestinal tract where haemorrhagic enteritis and intestinal perforation can occur ( martindale , 1993 ). animals were monitored daily for gi tract upset such as diarrhoea and weekly for more severe toxicity manifestations such as weight loss . weight loss was monitored by calculating net body weight as estimated by subtracting tumour weight , which was calculated as 1 g × tumour volume ( cm 3 ) as cited in shibamoto et al , 1996 . for demonstration of any weight changes the animal body weight was normalised to the body weight at the time of treatment commencement as no treatment toxicity was noted throughout the 6 - week study . in comparison to the 5 - fu treatment group the mice receiving ha therapy , that is as a sole agent or as a chemosensitizer , demonstrated enhanced well being where the animal did not loose weight , but maintained its body mass ( fig4 ). as one of the major toxicities associated with 5 - fu treatment is depression of the bone marrow and subsequent drop in white blood cells it was necessary to assess any treatment associated blood toxicity . upon anaesthetising the animals , blood was collected from the heart or great vessels using a needle and syringe . estimation of white blood cell number by making a 1 / 50 dilution of blood in mouse tenacity saline ( m ) and counting it on a haemocytometer . a differential blood count was performed by counting - neutrophils , lymphocytes , and erythrocytes . the total estimation of blood cell sub - populations was compared to published data for mouse blood . the total white cell count and sub - population differential were not significantly different , regardless of the treatment . to ensure that treatments did not induce organ atrophy or enlargement , the organs were removed and weighed during the post mortem . the mass of each organ was calculated as a % of the overall net body weight , and compared to the organ masses of the saline only group ( group 1 ). the overall patient survival time was calculated as the time ( days or weeks ) that the animal lived after the commencement of treatment . all animals in each treatment group completed the 6 - week treatment program . in relation to organ mass , the ha therapy did not result in any dramatic toxicity . mice receiving 5 - fu exhibited an enlarged spleen ( 61 % increase in mass ), while the co - administration of ha and 5 - fu significantly counteracted this enlargement by 31 % ( student t - test , p & lt ; 0 . 001 ). the 5 - fu therapy resulted in a shrinkage of the uterus ( 22 %), once again the ha / 5 - fu therapy reduced this toxic effect by 10 % ( student t - test , p = 0 . 04 ). it was also clearly defined that the addition of ha to the treatment regimen , when co - administered or administered the day before , significantly decreased the primary tumour mass in comparison to the saline treatment group ( student t - test , p = 0 . 006 ). no other differences in organ mass were noted between treatments . mda - mb 468 , mda - mb 435 and mda - mb 231 cells were cultured as described in example 2 . when the cultures had reached 70 - 80 % confluency they were washed in 1 × hbss at 37 ° c . and trypsinised in 10 ml of 0 . 25 % trypsin / 0 . 05 % edta until cells have fully detached . after add 1 ml of fcs to neutralise trypsin the cells were counted , centrifuged at 1 , 200 rpm for 5 min and resuspended as follows : cells were then plated into 48 - well plates and incubated in accordance with suppliers &# 39 ; instructions . after 24 h media was removed and replaced with the following test media : 40 nm adriamycin media : 450 ml ( stock adriamycin is 1 . 7 mm , therefore 1 , 700 , 000 / 40 = 42 , 500 ; 450 , 000 / 42500 = 10 . 6 ul of 1 . 7 mm adriamycin + 450 ml media ). this study has definitively proven that ha , can enhance the cytotoxicity of anti - cancer drugs , 5 - fu and adriamycin , both in vitro and in vivo . more specifically : 1 ). as a sole agent ha can exert a cytotoxic effect on cancer cells both in vitro and in vivo ( fig5 ); 2 ). evaluation of the therapeutic efficacy of ha sole therapy or chemosensitizing therapy demonstrated that it was not toxic to normal tissue and it did not enhance the toxicity profile of the drug . in fact , mice receiving the therapy displayed a significant weight gain over the 6 - week treatment period and a reduction in lymph node metastasis . the co - administration of ha and 5 - fu had a dramatic effect on the reduction of the primary tumour volume ; and 3 ) mice who had ha incorporated into the treatment regimen did not display the formation of any secondary tumour ( fig6 ). experiments are presently being conducted on the use of ha for in vivo treatment of breast cancer . these experiments are focusing on the effect of ha concentration and molecular weight and on the cytotoxicity of adriamycin . it is the aim of these studies to also establishing drug and ha exposure time and regimens , as well as the mechanism of action of ha , ie : receptor mediated transport and / or effect on cell membrane . further data on the role of ha in chemosensitizing drug - resistant cancer cells will also be collected . all studies will be conducted on breast cancer cell lines that express differing levels of ha receptors , cd 44 and rhamm . cell lines to be tested , mda - mb 435 , mda - mb 231 , mda - mb 468 , zrl - 751 and several mdr - 1 expressing breast cancer cell lines . investigation of the effect of ha / adriamycin exposure times and concentration on drug - resistant and drug - sensitive breast cancer cells . four mdr - 1 positive and 4 mdr - 1 negative cell lines will be exposed to adriamycin at 1 , 2 . 5 , 5 , 10 , 20 , 40 , 60 , 80 and 100 nm , the following variables will be tested : 1 ). 1 h drug ± 100 nm ha exposure followed by 3 days of drug - free growth ; 2 ). constant drug exposure ± 100 nm ha for 3 days 30 min 100 nm ha exposure , followed by drug for 1 h , cells grown drug - free for 3 days ; and 3 ). 24 h 100 nm ha exposure , followed by drug for 1 h , cells grown drug - free for 3 days . these experiments will establish ; optimal ha exposure times and regimens , magnitude of increased adriamycin cytotoxicity when combined with ha and whether ha can overcome efflux pump resistance in breast cancer cells . to date the ic 50 of adriamycin has been determined as 90 nm using 90 nm of adriamycin the ha ( 700 kd ) concentration will be varied to 1 , 3 , 10 , 30 , 100 , 300 nm , 1 μm , 3 μm , 10 μm , 30 μm and 100 μm . the incubation variables to be tested are : 1 ). 30 min ha exposure followed by 1 h drug exposure cells grown drug - free for 3 days ; 2 ). 24 ha exposure followed by 1 h drug exposure cells grown drug - free for 3 days ; and 3 ). ha ± drug exposure for 1 hr , cells grown drug - free for 3 days . any detached cells will be tested for cell viability since it has been suggested that ha can play a pivotal role in cancer cell detachment and migration . if detached cells are viable the ha receptor status will be determined using facs surface epitope identification . similar experiments will be performed with short ha oligiosaccharides , ie : 4 sacc , 6 sacc , 12 sacc , 5600 da , 50 , 000 da , 100 , 000 da , 250 , 000 da . these experiments will demonstrate the optimal ha : drug ratio in vitro , optimal ha exposure time and regimen , effect of ha molecular weight on adriamycin cytotoxicity . after determining the optimal ha concentration , the ic 50 of adriamycin will be used in a series of time course experiments to observe any effect of ha on adriamycin metabolism . the [ 14 c ] adriamycin will be exposed to the cells for 30 min , 1 h , 2 h , 4 h , 8 h , 16 h and 24 h . the experimental conditions will be : 1 ). exposure of cells to ha for 30 min followed by drug ; and 2 ). exposure of cells to ha for 24 followed by drug co - exposure of ha / adriamycin . cells will be removed , hypotonically lysed and centrifuged at 113 , 000 gav for 1 hr . the membrane pellet and supernatant will be counted and analysed for metabolites using hplc . cells will also be grown on coverslips , where they will be exposed to adriamycin ± ha ( exposures regimen as above ) and a confocal photography time course will be used to track the cytosolic uptake and movement of the drug . identification of ha receptors on mdr - 1 positive and negative breast cancer cell lines , facs quantitation of the cd44s , cd44v6 , cd44v10 and rhamm receptors will be conducted . quantitation of the ha / receptor binding and saturation kinetics using fitc / ha and facs analysis will also be done . we will be able to determine any of these block cd44s and rhamm receptors . the receptor status of any viable cells will be quantitated using surface epitope facs analysis . if blocking of the ha receptors decreases the normally observed synergism between adriamycin and ha , the membrane bound and cytosolic adriamycin will be quantited ± ha receptor blocking . ha degradation by cell lines using [ 3 h ] ha and gel filtration chromatography ± receptor blocking will be studied . ha of molecular weight , 4 sacc , 6 sacc , 12 sacc , 5600 da , 50 , 000 da , 100 , 000 da , 250 , 000 da , 750 , 000 da and 1 , 500 , 000 da will be incubated with breast cancer cell lines at pre - determined “ observed - effect ” concentrations and the following will be parameters investigated : extracellular and intracellular calcium flux ( cellular probe assays ). regulation of cytoskeletal components ( micro - array of cytoskeletal genes ), effect on volume of cells ( coulter size analysis ) and mobility of cancer cells ( boyden chamber matrigel assays ) will also be conducted . the effect of ha on the cell cycle will be undertaken by incubating ha of molecular weight , 4 sacc , 6 sacc , 12 sacc , 5600 da , 50 , 000 da , 100 , 000 da , 250 , 000 da , 750 , 000 da and 1 , 500 , 000 da with breast cancer cell lines at pre - determined “ observed - effect ” concentrations . cells will be labelled with potassium iodide and subjected facs analysis . the number of cells in each stage of the cell cycle will be determined . comparisons of the in vitro efficacy of the liposomal doxorubicin and ha / doxorubicin preparations will be conducted using the optimal ha / doxorubicin preparation and the dosage range used by the liposome company in the pre - clinical testing of the liposomal doxorubicin . before progression of the ha / adriamycin anti - cancer therapy into phase i human breast cancer trials it is necessary to conduct preliminary toxicity experiments . the experiments will focus on : 1 ). effect of hyaluronan on adriamycin uptake in mouse body organs and fluids ; 2 ). establish a preliminary dose range for adriamycin determine if ha targets adriamycin to human breast tumour xenografts in nude mice ; 3 ). compare the commercial liposomal doxorubicin to ha / doxorubicin uptake in mice ; and from inaba et al , ( 1988 ) the dose of adriamycin in nude mice was 4 mg / kg which is a human - equivalent dose of 60 mg / m 2 . nude mice bearing human tumours will be injected with adriamycin ± ha . using adraimycin concentrations of 4 mg / kg ± 12 . 5 mg / kg ha . the experimental protocol will include the following treatment groups : using adriamycin ± ha will be quantitatively injected into the tail vein of the mouse . at the time intervals of 2 , 15 , 30 , 60 min and 1 . 5 , 2 , 4 , 8 , 24 and 48 h ( 4 animals / time point ) the mice will be killed by a 0 . 1 ml ip injection of nembutal . all body organs , skeletal muscle , lymph nodes , bone marrow , urine and blood will be removed and the adriamycin content determined using hplc and fluorescence . human breast tumours will be generated in nude mice ( wehi cba strain ) the mice will be injected with : the above mentioned will be quantitatively injected into the tail vein of the mouse ( 8 animals / group ) on days 2 , 4 , 6 of a weekly cycle . tumour volume , body mass , food intake and functionality of the mice will be monitored on a daily basis . at the completion of the 8 - week study the mice will be killed by a 0 . 1 ml ip injection of nembutal . all body organs , tumour , skeletal muscle , lymph nodes , bone marrow , urine and blood will be removed processed for pathological assessment . to answer some basic questions about the effect of ha anti - cancer therapy on colon cancer cells the following experiments should be conducted . investigation of the effect of ha / 5 - fu exposure times and concentration on drug - resistant and drug - sensitive colon cancer cells . three resistant and 3 sensitive cell lines will be exposed to 5 - fu at 1 , 2 . 5 , 5 , 10 , 20 , 40 , 60 , 80 and 100 nm , the following variables will be tested : 1 ). 1 h drug ± 100 nm ha exposure followed by 3 days of drug - free growth ; 3 ). 30 min 100 nm ha exposure , followed by drug for 1 h , cells grown drug - free for 3 days ; and 4 ). 24 h 100 nm ha exposure , followed by drug for 1 h , cells grown drug - free for 3 days . using the ic 50 of 5 - fu as determined as above , ha ( 700 kd ) concentration will be varied to 1 , 3 , 10 , 30 , 100 , 300 nm , 1 μm , 3 μm , 10 μm , 30 μm and 100 μm . the incubation variables to be tested : 1 ). 30 min ha exposure followed by 1 h drug exposure cells grown drug - free for 3 days ; 2 ). 24 ha exposure followed by 1 h drug exposure cells grown drug - free for 3 days ; and 3 ). ha ± drug exposure for 1 hr , cells grown drug - free for 3 days . any detached cells will be tested for cell viability since it has been suggested that ha can play a pivotal role in cancer cell detachment and migration . if detached cells are viable the ha receptor status will be determined using facs surface epitope identification . similar experiments will be performed with short ha oligiosaccharides , ie : 4 sacc , 6 sacc , 12 sacc , 5600 da , 50 , 000 da , 100 , 000 da , 250 , 000 da . after determining the optimal ha concentration , the ic 50 of 5 - fu will be used in a series of time course experiments to observe any effect of ha on adriamycin metabolism . the [ 3 h ] 5 - fu will be exposed to the cells for 30 min , 1 h , 2 h , 4 h , 8 h , 16 h and 24 h . the experimental conditions will be : 1 ). exposure of cells to ha for 30 min followed by drug ; and 20 . exposure of cells to ha for 24 followed by drug co - exposure of ha / 5 - fu . cells will be removed , hypotonically lysed and centrifuged at 113 , 000 gav for 1 hr . the membrane pellet and supernatant will be counted and analysed for metabolites using hplc . cells will also be grown on coverslips , where they will be exposed to 5 - fu ± ha ( exposures regimen as above ) and a confocal photography time course will be used to track the cytosolic uptake and movement of the drug . identification of ha receptors on resistant and sensitive colon cancer cell lines , facs quantitation of the cd44s , cd44v6 , cd44v10 and rhamm receptors , quantitation of ha / receptor binding and saturation kinetics using fitc / ha and facs analysis will be done . blocking of cd44s and rhamm receptors with inhibitory antibodies , apply 5 - fu ± ha following the protocols of : 1 ). exposure of cells to ha for 30 min followed by drug ; and 2 ). exposure of cells to ha for 24 followed by drug co - exposure of ha / 5 - fu . cells will be counted . the receptor status of any viable cells will be quantitated using surface epitope facs analysis . if blocking of the ha receptors decreases the normally observed synergism between 5 - fu and ha , the membrane bound and cytosolic 5 - fu will be quantited ± ha receptor blocking . ha degradation by cell lines using [ 3 h ] ha and gel filtration chromatography ± receptor blocking will be studied . effect of ha on the plasma membrane hyaluronan of molecular weight , 4 sacc , 6 sacc , 12 sacc , 5600 da , 50 , 000 da , 100 , 000 da , 250 , 000 da , 750 , 000 da and 1 , 500 , 000 da will be incubated with breast cancer cell lines at pre - determined “ observed - effect ” concentrations and the following will be parameters investigated : an investigation of the role of ha neo - adjuvant therapy on the inhibition of organ metastasis will be undertaken . in comparison to other treatment groups , mice receiving the ha therapy have demonstrated that : 1 ). reduced lymph node metastasis as compared to other treatment groups ; these results highlight the possible role of ha anti - cancer therapy as an efficient means of reducing the spread of cancer . through the obligatory choice of a pre - clinical model there is a restriction , whereby the spread of the secondary cancer normally occurs in the surrounding lymph nodes . it would be advantageous to use a model where we can examine the spread of the cancer to every organ and the bone . by using a model known as the bag vector metastasis model we would be able to monitor the spread of cancer to every organ and the bone . in brief , the bag vector consists of a neomycin - resistant lacz gene that can be stably transfected into human breast cancer cells . after intracardiac injections into the nude mice , followed by a 6 - week treatment program it is possible to pcr detect the lacz gene in any metastasizing cells / organs . faxitron scanning with detection of bone lesions would detect any bone metastasis . the below treatments will be administered on day 1 , day 2 of a weekly cycle , for 6 weeks . the treatment groups ( 5 animals per group ) will consist of : 10 . 12 . 5 mg / kg ha on day 1 , 15 mg / kg mtx on day 2 , 12 . 5 mg / kg ha on day 3 , 15 mg / kg mtx on day 4 ; 12 . 15 mg / kg mtx , 30 mg / kg 5 - fu , 30 mg / kg cyclophosamide on day 1 , day 2 ; and 13 . 15 mg / kg mtx , 30 mg / kg 5 - fu , 30 mg / kg cyclophosamide + 12 . 5 mg / kg ha on day 1 , day 2 ; 12 . 5 mg / kg ha on day 1 , ( 15 mg / kg mtx , 30 mg / kg 5 - fu , 30 mg / kg cyclophosamide ) on day 2 , 12 . 5 mg / kg ha on day 3 , ( 15 mg / kg mtx , 30 mg / kg 5 - fu , 30 mg / kg cyclophosamide ) on day 4 . mouse mass and well being will be monitored daily for 6 weeks . on completion of the treatment cycle , each mouse will be scanned for bone lesions . after scanning each organ and body fluid will be removed . a sufficient cross section of the organ will be kept for possible future pathological analysis , while the remaining tissue will be homogenized and subjected to competitive pcr for the detection of the lacz gene . any organs which exhibit metastasis will be histologically processed and the pattern of colonization of the cancer cells will be noted using galactosidase staining of the lac z gene . culty , m ., shizari , m ., erik , w ., thompson . and underhill , c . b . ( 1994 ). binding and degradation of hyaluronan by human breast cancer cell lines expressing different forms of cd44 : correlation with invasive potential . journal of cellular physiology 160 : pp 275 - 286 . culty , m ., nguyen , h a , and underhill , c b . ( 1992 ). the hyaluronan receptor ( cd44 ) participates in the uptake and degradation of hyaluronan . j cell biol 116 ( 4 ): pp 1055 - 1062 . lang f ., ritter m ., volkl h and haussinger d ( 1993 ). the biological significance of cell volume ren physiol biochem . 16 : pp . 48 - 65 . wang , c ., zhang , s . and turley , e a . ( 1996 ). the role of hyaluronan and hyaluronan receptors in breast cancer cell invasion , motility and proliferation . in : fourth international workshop on hyaluronan in drug delivery . ( editor : willoughby , d . a ) roy . soc . med . press . pp 37 - 53 . wang , c ., tammi , m ., guo , h . and tammi , r . ( 1997 ). hyaluronan distribution in the normal epithelium of esophagus , stomach , and colon and their cancers . american journal of pathology . 148 ( 6 ): pp 1861 - 1869 .

the present invention relates to the enhancement of bioavailability of chemotherapeutic agents for the treatment of disease . in particular , the present invention relates to a method of enhancing the bioavailability of a chemotherapeutic agent comprising the step of administering to a subject in need thereof a therapeutically effective amount of hyaluronan .

fig1 shows a schematic of the apparatus for vep recording using virtual reality goggles ( 1 ), which present the display to the subject . the goggles are connected to a computer ( 2 ) with a linked video board that generates the multifocal stimulus . recording electrodes on the scalp ( 5 ) and a ground reference electrode ( shown on the earlobe ), detect the vep signal from one or more recording channels ( in this case four channels are shown ). the signals are conducted to an amplifier ( 3 ), before being processed by software for presentation on the operators display ( 4 ). results can be compared for each eye , or between the two eyes of a subject , with respect to normal reference values . fig2 shows a schematic of the apparatus for multifocal erg recording using virtual reality goggles ( 1 ). the set up is the same as in fig1 except that the recording electrode is placed in contact with the eye or eyelid . a ground electrode is required ( shown on the earlobe ). only one channel recording is required for the erg . fig3 is an example of a multi focal multichannel vep recording from the right and left eye of a normal subject . fig3 a shows the responses achieved using a conventional screen ( 22 inch hitachi monitor ) to present the stimulus . a cortically scaled dartboard stimulus was generated with 60 different areas of pattern stimulation using the objectivision perimeter . the trace array shown in the figure represents the responses generated from each part of the visual field tested out to 27 degrees of eccentricity temporally and 34 degrees nasally . for graphics purposes the central areas are relatively enlarged to show the raw vep signal within that area . fig3 b shows a multifocal multichannel vep recordings from the same normal subject as in fig3 a , recorded using virtual reality goggles to present the same stimulus instead of the conventional monitor . the same objectivision system was used . the responses are of similar order of magnitude in the two techniques , although there is some variation in amplitude across the field . due to the specifications of the goggles used , the display was limited to 21 degrees temporally and 27 degrees nasally . fig4 provides a comparison between subjective perimetry findings and the objective vep assessment of the visual field using virtual reality goggles . fig4 a shows the grayscale and pattern deviation printout from a subjective humphrey visual field test of the right eye of a glaucoma patient . an inferior arcuate scotoma ( blind spot ) is shown in the visual field . fig4 b shows the multifocal multichannel vep recording from the same eye as in fig4 a , recorded using virtual reality goggles . analysis of the signals demonstrates loss of vep responses corresponding to the inferior scotoma in fig4 a , with more extensive reductions in the superior field than seen on the humphrey . the amplitude deviation plot shades areas according to probability of abnormality when compared to a reference range of normal values extrapolated from the conventional screen objectivision system . this suggests that the technique is capable of detecting visual field loss in glaucoma , just as it is with the use of the conventional large screen . it may also demonstrate more significant glaucomatous damage than suspected on conventional humphrey field testing . five glaucoma patients have been tested with the virtual reality goggles and the scotomas were detected in all five cases . examination of multifocal vep data from normal subjects using conventional crt monitors demonstrated that the amplitude of the multi - focal vep is not age - dependant ( contrary to most electrophysiology parameters , eg the pattern erg ). in fact , some elderly people produce vep responses of higher amplitude . individual variation in the thickness of the scalp or subcutaneous tissue may cause inter - individual differences in vep amplitude due to variable impedance of bone and fat . direct measurement of the thickness or impedance of these tissues is not currently practical . however , the impedance will also affect the amplitude of the spontaneous brain activity ( eeg ) in a similar fashion to the vep . to confirm this we conducted a study using the objectivision vep perimeter of the correspondence between spontaneous eeg amplitude ( 99 % confidence interval ) and multifocal vep amplitude ( largest amplitude of a trace ). the study included 34 normal subjects . the results demonstrated a strong correlation between the eeg amplitude and vep ( correlation coefficient r = 0 . 81 ). the scatterplot for the correlation is shown in fig5 . an alternative method to measure background eeg activity is to calculate a fourier power spectrum of the eeg . therefore , if the level of spontaneous eeg activity is calculated during the recording , it provides an indirect measure of the overall registration of brain signals for that individual for the electrode positions used . whilst it is recognised that eeg amplitude is determined by many additional factors other than conductivity , it is proposed that scaling of an individual &# 39 ; s vep responses according to their eeg levels , relative to normal population eeg values , helps to reduce inter - individual vep variability . the eeg amplitude is approximately 1000 × the amplitude of the vep , so it is reasonable to assume that the vep signals themselves will have little contribution to the raw eeg levels . in analysis of multifocal vep recordings the eeg raw data is actually examined by cross - correlation techniques to extract the vep signals . when recording from an individual , the overall level of the raw eeg ( 99 % confidence interval ) as recorded during each run of the vep recording , can be used to provide an individual &# 39 ; s scaling factor . the vep extracted is then scaled by the eeg scaling factor . the value of the technique of the invention of vep scaling was confirmed by examining the data from 50 normals . the coefficient of variation for all 60 visual field test points had a mean value of 50 . 1 %. when the results were scaled according to background eeg values the coefficient of variation for all 60 visual field test points was reduced to 28 . 2 %. by using eeg scaling , the sensitivity of the test was also improved . in a study of 60 glaucoma cases using the objectivision system for multifocal vep perimetry , several glaucoma cases were not flagged as abnormal using the unsealed data since the subjects had overall large signals compared with normal , even though focal relative reductions could be seen when examining the trace arrays . with the data scaled according to eeg levels however , these subjects were identified as having localised reductions in their vep amplitudes and the scotomas were flagged appropriately . the eeg raw data can contain a large component of alpha rhythm signals and also spikes of electrocardiogram signals . if these are not excluded from the scaling factor applied , then some subjects will have their data inadvertently scaled down lower than is appropriate . this can introduce false positive results in the vep . one technique for rectifying this problem is to examine the raw signal by fourier analysis and any alpha - rhythm spikes and electrocardiogram signals can be identified . these can then be excluded from the spectrum before calculating a scaling coefficient . therefore scaling of the vep amplitude based on amplitude of spontaneous brain activity eliminates part of the variability between individuals caused by differences in conductivity of tissues . this technique has application in analysing multifocal vep signals recorded with conventional crt monitors , plasma screens , lcd screens , or with virtual reality goggles . the method and system of this invention will find wide use in the medical field , specifically in the field of ophthalmology . the foregoing describes only some embodiments of the invention and modifications can be made thereto without departing from the scope of the invention . 1 . baseler h a & amp ; sutter e e . vis research 1997 ; 37 ( 6 ): 675 - 790 2 . klistorner a i , et al invest ophthalmol vis sci 1998 ; 39 ( 6 ): 937 - 950 3 . klistorner a i , et al aust n z j ophthalmol 1998 ; 26 : 91 - 94 . 4 . graham s l , & amp ; klistorner a . aust n z j ophthalmol 1998 ; 26 : 71 - 85 5 . graham s l , et al surv ophthalmol 1999 ; 43 ( suppl1 ): s199 - 209 6 . graham s l & amp ; klistorner a . curr opin ophthalmol 1999 ; 10 : 140 - 146 . 7 . graham s l , et al j glaucoma 2000 ; 9 , 10 - 19 8 . kondo , m , et al invest ophthalmol vis sci , 1995 ; 36 : 2146 - 2150 10 . johnson c a , et al j glaucoma 2000 ; 9 ( ags abstract ): 110 12 . graham s et al vol 40 invest ophthalmol vis sci , 1999 , 40 ( 4 ) arvo abstract # 318 15 . sarwate & amp ; pursley . proc ieee , 1980 , vol 68 ( 5 ) 593 - 619 17 . kamaletdinov b . problems of information transmission , 1988 , vol 23 ( 2 ) 104 - 107

for electrophysiological assessment of visual function using a head mounted stereo display for displaying a stimulus which is used to generate a retinal or cortical response . in particular , a method for objective electrophysiological assessment of visual function of at least one eye of a subject includes presenting a visual stimulus to at least one eye of the subject , recording at least one of a retinal response and , a cortical response generated as a result of the presenting ; analyzing said response and , as a result of said analyzing , forming a map of the visual function of the at least one eye of the subject 6 . the invention also relates to as system for such electrophysiological assessment .

referring to fig1 , a machine 10 is shown having a tractor 12 and a front aerator 14 . front aerator 14 may be mounted at the rear of tractor 12 or at the front of tractor 12 . the primary requisite of the aerator is that it be capable of providing an aeration of the entire windrow and also movement of the windrow 16 laterally while at the same time exposing the floor 26 of the shed 27 . several machines 10 are available for aerating and moving the windrow . a preferred machine is made by brown bear corporation , corning , iowa . fig2 - 7 show a typical shed 27 . referring to fig2 the litter 24 covers the entire floor 26 of shed 27 . the litter 24 can be wood chips , sawdust , or other material that is loose , but covers the entire floor 26 of shed 27 . a partial windrow 22 is formed by a first pass that removes the litter 24 from the floor 26 and places it upon the litter 24 adjacent the exposed floor 26 . referring to fig3 a first windrow 28 is formed by again passing the machine 10 along the partial windrow 22 and moving the windrow 28 and the litter 24 below it to the position shown in fig3 . this leaves an exposed floor 26 . then a partial windrow 30 and a partial windrow 32 are provided by passing the machine 10 at the outside walls of the floor as shown in fig4 . by making passes five and six it is possible to form the first windrow 34 , the first windrow 36 and the first windrow 28 as shown in fig5 . fig8 shows leaving of the windrow 28 in the position shown in fig8 for approximately 72 hours . during this time the center portion 50 achieves a temperature of 130 ° f . or greater . similarly , the temperature 52 at the outermost regions of the windrow 28 achieves a temperature that is less 130 ° f . the windrow 28 has a height 1½ to 3½ feet . similarly the width of the windrow 28 is approximately 6 to 9 feet . this defies earlier judgments that the greater the windrow height and width , the greater temperature would be achieved at hot spot 50 . however , it has been found that a height of 1½ to 3½ feet and a width of 6 to 9 feet results in a hot spot 50 that is greater than 130 ° f . the process as shown in fig6 shows an additional pass of aeration equipment 14 to create second windrows 38 , 40 , 42 . essential to the creation of second windrows 38 , 40 and 42 is an aeration of the first windrows 28 , 34 , 36 and also the moving to a portion at floor 26 to create the second windrows 38 , 40 , 42 . a third turning and aeration of the windrows as shown in fig7 creates a third windrow 44 , 46 , and 48 . similar to the encounter by machine 10 , the windrows 44 , 46 , 48 should be aerated and also should be removed from the floor 26 to a third position on floor 26 . referring to fig9 , a graph 54 , a graph 56 , and a graph 58 are shown . graph 54 shows a single windrowing wherein the windrow is not turned . graphs 56 and 58 show turning the windrow a second and a third time . there can be additional turnings of the windrows , and each turning shows additional pasteurization . however , a second or a third time are preferred . as can be shown in fig9 , the graph 54 quickly heats to in excess of 130 ° f . in approximately one day . graph 54 reaches a height of approximately 150 ° f . and then begins its descent to below 130 ° f . at approximately day 7 . the graph 54 continues its descent below 130 ° f . this can be shown in fig8 wherein the hot spot 50 represents the temperature which exceeds 130 ° f . however , the outside temperature 52 is less than 130 ° f . and requires additional turning of graphs 56 , 58 as shown in fig9 . graph 56 shows a reduction in the temperature of hot spot 50 , but quickly shows it increasing to a temperature exceeding 140 ° f . at the sixth day the material is turned again into a third windrow , and while initially there is a drop in the temperature between the sixth and seventh day , there is an increase thereafter . as can be seen in graphs 56 , 58 , the temperature of hot spot 50 continues to stay above 130 ° f . whereas the graph 54 shows a continuing decline . thus it is possible to turn the graph 54 between the fourth and sixth day as shown in graph 56 , and then turn the graph between six and nine days to create a third windrow . while graph 56 is sufficient to pasteurize all of the pathogens , additional windrowing of the third windrow 44 , 46 , 48 results in increased reduction of pathogens . among the factors that affect windrowing , is the bedding material . this is a source of carbon to compost high nitrogen poultry litter . it involves rice hulls , sawdust blend , shavings , or other materials . rice hulls are the best because they are available in carbon and are good particle size . shavings suffice because they are high in carbon , but they are sometimes not available , and some have large particle size . it may be possible to combine the rice hulls and the sawdust to compost the material . other materials may be used . another factor is the moisture in the litter . preferably the moisture in the litter should exceed 30 % by weight . 35 to 60 % is ideal , but 65 % or exceeding that is the least desirable . how the deep the litter is is also important . preferably there is less than 4 inches and they should be placed in two or three windrows in 36 to 43 foot wide houses . four to six inches is ideal . exceeding six inches should have some material taken to the litter shed for further composting or for composting mortalities . how long since the house was totally cleaned out to the floor is also important . after 4 - 6 sets of chickens the house should be cleaned out to the ground and then start composting the next flock . the time the grower has between flocks 21 to 28 days is ideal . more percentage of disease bacteria and pathogen is killed using this process and the nutrients are stabilized . this is best for beetle control . 20 - 11 days also will work . there is potential for ammonia and may need some litter amendment . some beetle control but not as good as 21 - 28 days . 11 - 4 days is still good . possible high ammonia and may need more litter amendment . good beetle control . windrows should be turned every three to five days for 21 - 28 days and then as needed to dry litter below the 35 % moisture area . beatles , mites , and disease bacteria and pathogens should be eliminated by this process . before windrowing poultry houses , all water and feed lines need to be raised as high as they can go . old cables , cords , box heaters , sorting tubes , trash and curtains should be cleaned out of the litter . the house should be washed or blown down so that bacteria and pathogens and bugs in the dust and cobwebs also can get composted so as to not reinfect the house . the advantages of turning windrows are that the turning assures aerates the litter and gets more of the windrow in the core heat zone . it also provides better pathogen reduction , and moves the windrow so that the floor will dry out under the windrow . flash off of steam or moisture and the ammonia gas is also achieved . the process breaks up chunks of the cake , and blends dry with wet materials . an advantage involves the trapping of beetles and beetle larvae in the core heat zone . in the drawings and specification there has been set forth a preferred embodiment of the invention , and although specific terms are employed , these are used in a generic and descriptive sense only and not for purposes of limitation . changes in the form and the proportion of parts as well as in the substitution of equivalents are contemplated as circumstance may suggest or render expedient without departing from the spirit or scope of the invention as further defined in the following claims .

this process comprises aerating chicken litter into a first windrow and aerating a second time into a second windrow . each of the windrows are permitted to achieve a temperature of approximately 130 ° f . internally . the litter is then spread to completely cover the entire area and a second set of chickens is placed on the litter . the more aeration times that the windrow is made the more the litter is pasteurized and the pathogens and beetles are removed .

referring now to fig1 and 2 , there are shown side and partially exploded perspective views , respectively , of a preferred embodiment of an access needle constructed according to the teachings of the present invention , said access needle being represented generally by reference numeral 11 . access needle 11 comprises a stylet assembly 13 and a cannula assembly 15 . referring now to fig1 through 3 , stylet assembly 13 comprises a stylet ( or sounding needle ) 17 and a stylet hub 19 . stylet 17 , which is preferably made of stainless steel , is a solid cylindrical member of small diameter , preferably 20 gauge , having a sharp , three - sided tip 21 at its distal end . hub 19 comprises a body 22 and a plug 23 . body 22 , which is preferably made of plastic , is a generally cylindrical hollow member having a proximal end wall 25 , a circular side wall 27 and an open distal end . the outer surface of side wall 27 is provided with a plurality of longitudinally extending ribs 29 to facilitate the gripping of body 22 . the inner surface of side wall 27 is provided with a helical thread 31 , the purpose of which will be described below . plug 23 , which is preferably made of plastic , is a generally cylindrical solid member fixedly mounted near its proximal end 33 within an opening ( not shown ) provided in end wall 25 , plug 23 extending coaxially with side wall 27 to define therebetween an annular space 28 . stylet 17 , which extends coaxially with plug 23 , is fixed in distal end 35 of plug 23 , preferably by insert - molding . referring now to fig1 , 2 , 4 and 5 , cannula assembly 15 comprises a cannula 45 and a cannula hub 47 . cannula 45 , which is preferably made of stainless steel , is a tubular member of small diameter , preferably 18 gauge , that is appropriately dimensioned for stylet 17 to be removably inserted thereinto . a circumferential band 51 , which may be produced , for example , by chemical - etching or the like , extends proximally a short distance ( e . g ., approximately 3 / 10 inch or more ) from the distal end 53 of cannula 45 . band 51 , which is readily distinguishable in appearance , e . g ., by color , from stylet 17 , permits facile differentiation of stylet 17 and cannula 45 , which is desirable for reasons to become apparent below . cannula hub 47 , which is preferably made of plastic , is a unitary member shaped to include a distal portion 61 , an intermediate portion 63 , and a proximate portion 65 . a plurality of longitudinal ribs 62 are formed on the outer surface of distal portion 61 to engage a protective sleeve ( not shown ) removably mounted over cannula 45 and stylet 17 when needle 11 is not in use . a plurality of laterally extending gripping elements 64 are formed on the outer surface of intermediate portion 63 to facilitate the gripping of hub 47 by a user . proximate portion 65 is appropriately dimensioned to be received in annular space 28 of stylet hub 19 . threads 66 are formed on the outside surface of proximate portion 65 and are adapted to engage thread 31 , thereby permitting hub 19 and hub 47 to be screwed ( or twist - locked ) together so as to prevent undesired relative longitudinal movement between stylet 17 and cannula 45 . a longitudinal bore 67 is formed in hub 47 , bore 67 having a distal region 69 disposed in distal portion 61 , an intermediate region 71 disposed in intermediate portion 63 and a proximal region 73 disposed in proximate portion 65 . distal region 69 is appropriately dimensioned to securely receive therewithin the proximal end of cannula 45 . proximal region 73 has a comparatively large diameter to facilitate the insertion of stylet 17 thereinto , and intermediate region 71 tapers in diameter from proximal region 73 to distal region 69 so as to facilitate the insertion of stylet 17 into cannula 45 . stylet 17 and cannula 45 are dimensioned so that , when hub 19 and hub 47 are coupled together in the twist - lock manner described above , stylet 17 extends distally beyond distal end 53 of cannula 45 for a sufficient distance so that , as will hereinafter be described , tip 21 of stylet 17 may be inserted into a jejunum , without also inserting cannula 45 thereinto , and a snare may be tightened around the thus inserted portion of stylet 17 . in the present embodiment , stylet 17 extends approximately 1 . 5 inch beyond distal end 53 of cannula 45 , cannula 45 having a length of about 2 . 5 inches . ( it should be understood , however , that the above dimensions of stylet 17 and cannula 45 may be varied . for example , stylet 17 and cannula 45 could be dimensioned so that cannula 45 has a length that is about ⅔ the length of stylet 17 .) the use of access needle 11 in the implantation of a pej tube in a patient in accordance with the teachings of the present invention will now be described . referring now to fig6 , after properly administering an anesthetic to a patient , an endoscope 8 having an illuminating channel is inserted through the patient &# 39 ; s mouth and into the digestive tract of the patient until it reaches the patient &# 39 ; s jejunum . scope 8 is used to transilluminate the jejunum through the abdominal wall , thereby identifying the location of the jejunum to a surgeon in order to facilitate the surgeon &# 39 ; s insertion of access needle 11 through the abdominal wall 10 and into the jejunum 12 of the patient . referring now to fig7 , the distal end of stylet 17 , but not cannula 45 , is then inserted through abdominal wall 10 and into jejunum 12 until it is visible to endoscope 8 through an observation channel therein . a snare 14 , which is inserted into the jejunum 12 through a snare channel of endoscope 8 , is then used to securely capture the inserted end of stylet 17 and to couple stylet 17 to snare 14 . with stylet 17 and snare 14 thus coupled , stylet 17 is pulled proximally so as to engage the wall of jejunum 12 . further pulling of stylet 17 results in jejunum 12 being held stationary against abdominal wall 10 . referring now to fig8 , with jejunum 12 held stationary against abdominal wall 10 by snare 14 and stylet 17 , cannula 45 is de - coupled from stylet 17 ( by unscrewing hub 19 from hub 47 ), and the distal end 53 of cannula 45 is then moved distally until it penetrates jejunum 12 through the opening previously created by stylet 17 . referring now to fig9 , snare 14 is loosened slightly from the inserted portion of stylet 17 and is then moved from stylet 17 to the inserted portion of cannula 45 . snare 14 is then tightened around the inserted portion of cannula 45 . referring now to fig1 , with snare 14 now tightened around cannula 45 , stylet 17 is then withdrawn proximally from cannula 45 . at this point , it is the combination of cannula 45 and snare 14 that hold jejunum 12 stationary against abdominal wall 10 . with stylet 17 removed , the distal end of an insertion wire 16 ( or a suture ) is inserted through cannula 45 and into jejunum 12 . snare 14 is then moved from around the inserted portion of cannula 45 to capture the distal end of insertion wire 16 . with snare 14 holding onto the distal end of insertion wire 16 , snare 14 and the distal end of insertion wire 16 are then withdrawn from the patient through the jejunum , the stomach and , ultimately , the mouth of the patient . referring now to fig1 , a pej tube 18 is then attached to the distal end of insertion wire 16 , which has previously been pulled out of the patient &# 39 ; s mouth . the proximal end of insertion wire 16 is then pulled proximally , causing pej tube 18 to be pulled through the patient until its proximal end extends out through the abdominal wall of the patient and its distal end is disposed within the jejunum of the patient . referring now to fig1 , the distal end of pej tube 18 can be seen to have an enlarged end , which serves to anchor the distal end of pej tube 18 in jejunum 12 . an external bolster 20 may be positioned over pej tube 18 to hold pej tube 18 in place on the patient . with pej tube 18 thus implanted , liquid nutrition materials and / or medications may be fed into the jejunum through pej tube 18 . it should be appreciated that , instead of positioning the pej tube in the patient by the pull - method as described above , the pej tube could alternatively be positioned in the patient using the push method . it should also be appreciated that the foregoing method and device could also be used to implant a peg tube in a patient or to implant other types of devices elsewhere in a patient . the present method and device could also be used , more generally , to position an organ within the body . the embodiments of the present invention described above are intended to be merely exemplary and those skilled in the art shall be able to make numerous variations and modifications to it without departing from the spirit of the present invention . for example , although stylet 17 is described herein as being a solid member , stylet 17 could instead be hollow . in addition , although cannula 45 is disclosed herein as having a blunt distal end 53 , distal end 53 could instead be tapered . moreover , whereas hubs 19 and 47 are described herein as being secured by a twist - lock , other removable securing means , such as a latch or a clip , could be used . also , instead of using a band to differentiate stylet 17 and cannula 45 , stylet 17 and cannula 45 could be colored differently , stylet and / or cannula 45 could be provided with graduated depth markers , or stylet and / or cannula could be provided with surface markings , textures or patterns . it should also be understood that the diameters of stylet 17 and cannula 45 could be varied . in addition , barbs or like means could be provided on the distal end of stylet 17 and / or cannula 45 to facilitate the grasping thereof by the snare . furthermore , various types of coatings ( e . g ., antithrombotic , antimicrobial , hydrophilic or hydrophobic ) could be applied to access needle 11 . all such variations and modifications are intended to be within the scope of the present invention as defined in the appended claims .

method for implanting a percutaneous endoscopic jejunostomy tube in a patient and access needle for use in the method . in one embodiment , the method comprises the steps of providing an access needle wherein the distal end of the stylet extends distally for an appreciable distance beyond the distal end of the cannula ; inserting the stylet , but not the cannula , into the jejunum ; grabbing the stylet with an endoscopically - positioned snare ; anchoring the jejunum against the abdominal wall using the snared stylet ; loosening the snare slightly while advancing the cannula into the jejunum and into the loosened snare ; tightening the snare around the cannula ; removing the stylet from the cannula ; inserting a guide wire or suture through the cannula into the jejunum ; and proceeding in the conventional fashion to implant a pej tube into the patient using the guide wire or suture .

referring now to the figures of the drawings in detail and first , particularly to fig1 thereof , there is shown a frame 1 that has a base part 2 that is , at the same time , a filter and corner stays 3 connected thereto . on a side opposite the base part 2 , the corner stays 3 are connected to a frame part 4 . the corner stays 3 can be made as separate parts and , then , attached to the base part 2 and connected to the frame part 4 . but , as shown in the embodiment , it is also possible to have the base part 2 , the corner stays 3 , and the frame part 4 be a monobloc configuration . in the corner regions of the frame 1 — in the illustrated embodiment in two corner stays 3 of the frame part 4 — receptacles 5 and 6 are formed . for example , a non - illustrated container that can be filled with a washing and / or cleaning agent can be incorporated into one receptacle 5 and a device for water softening 7 can be built into the other receptacle 6 . the frame 1 has wall openings 8 , which are configured as receptacles and which can , in turn , be sealed by separate wall parts 9 and 10 . the receptacles 8 fully peripherally enclose the wall parts 9 and 10 and , optionally , also the base 2 and / or lid parts attached to the frame 1 . the wall parts 9 and 10 , of the illustrated embodiment are connected suitably to the frame 1 , e . g ., by adhesion , welding or the like . of wall parts 9 and 10 , in the illustrated embodiment , one wall part 9 is configured as a simple wall part having only a closing function . in comparison , the other wall part 10 is configured as a heat exchanger . it is usual in dishwashing machines to utilize heat exchangers for economizing on power , through which preheating of the washing liquid utilized in rinse cycles following an introductory rinse cycle takes place , or that are filled in a drying cycle with cold water , to create large condensation surfaces . through integration of heat exchangers in one or several wall parts 10 of the washing container , there is practically no additional space requirement and a higher heat recovery and / or improved drying can be achieved from configuring several wall parts 10 as heat exchangers . if the washing container is a washing container that can be withdrawn from the housing of a dishwashing machine , then slide grooves 11 can also be provided on the frame 1 , with which the washing container can , then , be slid on slide rails available on the housing of the dishwashing machine . if appropriate , the modular structure of the washing container also allows the use of wall , base , and / or lid parts 2 , 9 , 10 being of various materials , for example , plastic or metal . if the base part 2 , the corner stays 3 , the frame part 4 and , optionally , a non - illustrated lid part that can be set onto the frame part 4 are configured as separate parts , then washing containers can be manufactured easily with different dimensions . because the individual parts exhibit relatively simple forms , namely , rod or plane form , they are also easy and cost - effective to manufacture . the invention has succeeded , therefore , in configuring a washing container that can easily be adapted to varying predefined requirements according to the configuration of the dishwasher .

a washing container for a dishwasher having several parts joined together , including a frame with walls , a base , and / or a cover attached thereto . the container can easily be adapted to varying predefined requirements according to the design of the dishwasher .

the waste is loaded into a housing defining a chamber ( 1 ) of strong sheet metal with sides protected by anti - wear shields , preferably made of manganese steel . the chamber is closed by a cover ( 2 ) manually or hydraulically actuated by means of a cylinder ( 22 ) and a hydraulic unit ( 24 ). inside the chamber , a vaned rotor ( 3 ), with highly resistant replaceable steel blades ( 4 ), is rotated at high speed , for example at 1500 rpm . the rotor is actuated by a powerful electric motor ( 5 ) and a system of belts and pulleys ( 6 ) tensioned by a belt tightener ( 7 ). a hydraulic coupling ( 23 ) for reducing absorption on starting and for compensating the forces owing to breakdowns or overloading can be mounted on the large - scale machines . as an alternative to the electrical system , actuation can be achieved by a hydraulic motor and an oleodynamic unit . preferably , sturdy , stationary , steel striker plates 8 , of which the purpose is to strike , crush and restrain the material such that the kinetic energy can be converted into heat energy , are mounted in the lower part of the chamber . preferably , the rotary shaft carries a central mounting suitable for receiving a test - tube carrier ( 9 ) with holes which can be opened and charged with samples of particular spores or bacteria ( such as , for example , bacillus stearothermophilus or bacillus subtilis ) such that the correct functioning of the sterilizing or disinfecting machine can be checked periodically in a suitable test cycle . at the end of the cycle , the material is discharged by the opening of a shutter ( 10 ), by means of a hydraulic system ( 11 ) or another motorized device . the apparatus is provided with safety devices to protect against accidents , and in particular with a microswitch device ( 12 ) which prevents the rotor starting if the cover is not closed , and a hydraulic device consisting of an automatic pump ( 13 / 1 ) and of a piston ( 13 / 2 ) which prevent the cover opening when the rotor is rotating . the temperature inside the chamber is measured and controlled by an indicator -- regulator ( 14 ), the sensitive bulb of which can be installed on the cover but preferably inside a stationary vane which actuates the opening of a solenoid valve ( 16 ) and the injection of water from a nozzle ( 15 ) in specific stages of the program . the emissions which are released during the treatment cycle pass into a small furnace ( 17 ) in which a high temperature ( approximately 800 ° c .) is achieved by electrical resistances such that these emissions are sterilized . after passing through the furnace , the emissions are absorbed in a water absorber with a nozzle ( 18 ) which is actuated by a solenoid valve ( 19 ). at the end of the step during which the temperature is maintained , the treated material is cooled by slowing down the speed of the rotor , by the dosing of the water , by placing the housing under a slight vacuum , by increasing the water at the nozzle ( 18 ), and by circulating air in a space ( 20 ) surrounding the housing , by means of a fan ( 21 ), or water driven by a motorized valve . the apparatus is completed by a hydraulic unit ( 24 ) for actuating the service pistons and valves , as well as an electric control panel ( 25 ). apparatus provided with a chamber which is 600 mm in diameter and 800 mm high is equipped with a coaxial , bladed rotor rotating at 750 - 1500 rpm and consisting of two blade - carrying vanes connected to a 50 kw electric motor . anti - wear shields and six stationary blades are disposed along the lower circumference of the chamber . 30 kg of infected hospital waste and 7 kg of a coloured polythenic master are introduced into the chamber and the cover is closed . the machine is started according to a programmed cycle in which a temperature of 160 ° c . is reached and maintained . the temperature is reached in 5 minutes , after which the water regulating and injecting system comes into action and maintains the temperature at this level for approximately 30 minutes . at the end of this step , the speed of the rotor automatically drops to 750 rpm and the dosing of the water decreases the temperature to 80 ° c . in 2 minutes . the cooling step is started by air being blown onto the jacket for 10 minutes ; thereafter , the temperature of the material is 60 ° c ., the opening of the shutter causes the granulated and sterilized material to be discharged automatically in approximately 1 minute . the machine then stops and is ready to perform a new cycle . apparatus provided with a chamber which is 1200 mm in diameter and 800 mm high is equipped with a coaxial , bladed rotor rotating at between 0 and 1500 rpm and consisting of two blade - carrying vanes connected to a 250 kw hydraulic motor . anti - wear shields and 25 stationary blades are disposed along the lower circumference of the chamber . 120 kg of infected hospital waste and 30 kg of coloured polypropylenic master are introduced into the chamber and the cover is closed . the machine is started according to a programmed cycle of which the first step is to reach a temperature of 150 ° c . this temperature is reached in 10 minutes , whereupon the water regulating and injection system comes into action and maintains the temperature at this level for 20 minutes . at the end of this step , the dosing of water is automatically suspended since the second step of the cycle program provides for the further heating of the mass to a temperature of 180 ° c . which is reached in 1 minute . the speed of the rotor then drops automatically to 500 rpm and the dosing of water decreases the temperature to 80 ° c . in 1 minute . the cooling step is started under a vacuum and by air being blown onto the jacket for 10 minutes , after which the temperature of the material is 50 ° c . the opening of the shutter causes the granulated and sterilized material to be discharged automatically in approximately 2 minutes . the machine then stops and is ready to perform a new cycle . in a further embodiment of the invention , a concentrated sodium hypochlorite acqueous solution is injected onto the mass of waste , within the housing during the sterilization cycle . thus the water , which is sprayed and dosed , during the cycle , may advantageously be in solution with sodium hypochlorite , preferably at a concentration of 12 - 15 % wt . active chlorine . it has been found that the concentrated sodium hypochlorite solution , at the high temperature reached within the housing , reacts with carbon dioxide to form undissociated hypochlorous acid which in turn dissociates to gaseous chlorine monoxide ( cl 2 o ) which is an effective chlorinating and biocidal agent . in this embodiment the waste material being comminuted is sprayed with the concentrated naclo solution when heated to a temperature higher than 150 ° c . and air , including carbon dioxide , is introduced within the housing . the amount of co 2 inherently present in the air is sufficient to cause chlorine monoxide evolution , which under the strong stirring conditions penetrates within the bulk of the waste being treated and flows through the apparatus thus improving the sterilization effect . residual amounts of chlorine monoxide , still present at the end of the treatment , if any , may be absorbed in the water absorber to give acqueous hypochlorous acid . thus , not only the waste is further sterilized , but also the emissions formed during the treatment and the internal surfaces of the apparatus . in this embodiment there is no need for feeding the emissions to furnace ( 17 ) prior to feeding them to the water absorber . naturally , the principle of the invention remaining the same , the forms of embodiment and details of description can be varied widely with respect to what has been described and illustrated above purely by way of non - limiting example .

a method for heat sterilizing or disinfecting hospital waste includes the steps of grinding and / or comminuting the waste under a cutting force and for an amount of time such that a quantity of heat which is sufficient to reach and maintain the sterilization or disinfection temperature within the massive waste is generated by friction . the apparatus includes a rotor provided with vanes or blades for grinding or comminuting the waste and a plurality of fixed strikers in the form of vanes or blades associated with the walls of the housing .

referring to the drawings and particularly to fig3 and 4 , one form of the stethoscope head covering apparatus of the present invention is there shown and generally designated by the numeral 12 . apparatus 12 here comprises a plurality of cover dispensing units 14 of the character illustrated in fig2 of the drawings and a feed unit 16 of the character shown in fig3 and 4 of the drawings . in a manner presently to be described , feed unit 16 functions to sequentially dispense the plurality of cover dispensing units 14 . however , it is to be understood that , as previously described , the cover dispensing units 14 can be packaged and distributed as individual handheld units that can be used by the physician to interconnect the protective cover with the stethoscope head . as best seen in fig1 and 2 of the drawings , each cover dispensing unit 14 comprises a positioning device 18 that includes a base 20 having a generally planar portion 22 and an upstanding rim 24 that is connected to portion 22 in the manner shown in fig1 . for a purpose presently to be described , generally planar portion 22 of the base is provided with an open segment 23 . similarly , upstanding rim 24 is provided with an open segment 24 a that is disposed in alignment with open segment 23 of the planar portion of the base . connected to each of the cover - positioning devices 18 to form a dispensing unit 14 ( see fig2 ) is a cover assembly 26 that comprises a uniquely constructed disposable cover 28 . as best seen in fig1 , disposable cover 28 has a thin , yieldably deformable , membrane - like central portion 28 a and a peripheral portion 28 b . connected to peripheral portion 28 b is an elastomeric bead 30 that is removably receivable over the upstanding rim 24 of the base 20 in the manner shown in fig2 of the drawings . considering once again the dispenser unit 16 of the apparatus of the invention , as illustrated in fig3 and 4 of the drawings , dispenser unit 16 here comprises a base 32 having a forward portion 32 a and a rearward portion 32 b . as indicated in fig3 of the drawings , forward portion 32 a is provided with a generally key - hole - shaped opening 34 , the purpose of which will presently be described . connected to the rearward portion 32 b of base 32 is an upstanding dispensing tower 36 having an interior chamber 36 a ( fig5 ) that , in the manner shown in fig5 , functions to hold a plurality of stacked cover dispensing units 14 . for a purpose presently to be described , the chamber is provided with a slot 36 a that is formed proximate the rearward portion 32 b of the base ( see fig5 ). slidably receivable within slot 36 a for movement between a first retracted position and a second dispensing position is an ejector slide 38 . when the selector slide has been moved from its first retracted position to its second dispensing position , a selected one of the cover dispensing units 14 , such as the device identified in fig3 and 4 by the numeral 14 x , has been moved along the base 32 from the inward position shown in fig3 into the outward position shown in fig4 and 5 wherein it resides over the generally key - hole - shaped opening 34 . with the cover dispensing units designated as 14 x in position over the generally key - hole - shaped opening 34 ( fig5 ), the disposable cover assembly 26 can be interconnected with the stethoscope head in the manner illustrated in fig5 , 5 a and 5 b of the drawings . more particularly , in order to interconnect the disposable cover with the stethoscope head , the stethoscope head is moved in the direction of the arrow 41 of fig5 a from the position shown in the phantom lines in fig5 into the downward position shown by the phantom lines in fig5 a , where it is in engagement with the yieldably deformable central portion 28 a of the cover . a continued downward movement of the stethoscope head in the direction of the arrow 41 of fig5 a will cause a deformation of the central portion of the cover in the manner shown in fig5 a . a further downward movement of the stethoscope head to the position shown in fig5 b will cause it to move through the generally key - hole - shaped opening 34 ( see fig3 ) and into the position shown in fig5 b of the drawings . as the stethoscope head moves downwardly through the keyhole shaped opening , the air - filled hollow tubes “ t ” of the stethoscope will pass through the segmented openings 23 and 24 a formed in the base 20 and in the upstanding rim 24 , respectively . as the stethoscope head moves downwardly , the elastomeric bead 30 will release from the rim portion of the base and , in the manner illustrated by the arrows 43 and fig5 a , will move into engagement with the peripheral portion “ p ” of the stethoscope head , thereby removably securing the cover 28 to the stethoscope head and moving the flexible central membrane portion 28 a of the cover into close engagement with the lower surface “ s ” of the stethoscope head . once the cover is in position over the stethoscope head , the assemblage thus formed can be removed from the apparatus 12 in the manner illustrated in fig5 c . following the examination of the patient , the cover assembly 26 can be conveniently removed from the stethoscope head and discarded into a suitable contaminated waste receptacle . turning next to fig6 through 10 , an alternate form of the stethoscope head covering apparatus of the present invention is there shown and generally designated by the numeral 42 . apparatus 42 , which is similar in many respects to the embodiment shown in fig1 through 5 , here comprises a plurality of cover dispensing units 44 of the character illustrated in fig8 and 9 of the drawings and a feed unit 46 of the character shown in fig6 and 7 of the drawings . in a manner presently to be described , feed unit 46 functions to sequentially dispense the plurality of cover dispensing units 44 . however , it is to be understood that , as previously described , the cover dispensing units 44 can be packaged and distributed as individual handheld units that can be used by the physician to interconnect the protective cover with the stethoscope head . as best seen in fig8 through 10 of the drawings , each cover dispensing unit 44 comprises a positioning device 48 that includes a positioning base 50 having a generally planar portion 52 ( fig8 ) and a rim 54 that is connected to positioning base 50 in the manner shown in fig1 . for a purpose presently to be described , generally planar portion 52 of the positioning base is provided with an open segment 53 . similarly , upstanding rim 54 is provided with an open segment 54 a that is disposed in alignment with open segment 53 of the positioning base . connected to each of the cover - positioning devices 48 to form a dispensing unit 44 ( see fig8 ), is a cover assembly 56 that comprises a uniquely constructed disposable cover 58 . as best seen in fig9 disposable cover 58 has a thin , yieldably deformable , membrane - like central portion 58 a and a peripheral portion 58 b . connected to peripheral portion 58 b is an elastomeric bead 60 that is removably receivable over rim 54 of the base 50 in the manner shown in fig9 and 11 of the drawings . considering once again the dispenser unit 46 of the apparatus of the invention , as illustrated in fig6 and 7 of the drawings , dispenser unit 46 here comprises a base assembly 62 that includes a base 63 having a forward portion 63 a and a rearward portion 63 b . a novel feature is illustrated in fig7 where the base assembly 62 can be attached to a vertical wall by fasteners “ s ” and anchors “ c ”. as indicated in fig7 of the drawings , rearward portion 63 b is provided with a feed unit receiving chamber 65 , while forward portion 63 a is provided with a generally key - hole - shaped opening 64 , the purpose of which will presently be described . receivable within feed unit receiving chamber 65 is the previously identified feed unit 46 that includes an upstanding dispensing tower 66 having an interior chamber 66 a ( fig7 ) that , in the manner shown in fig6 and 7 , functions to hold the plurality of stacked cover dispensing units 44 . as best seen in fig7 , to insure correct alignment of the feed unit 46 , base 63 of base assembly 62 is provided with a pair of index slots 63 c that closely receive a pair of indexing protuberances 46 a formed on feed unit 46 ( fig6 ). for a purpose presently to be described , base 63 is also provided with a guide channel 67 that communicates with feed unit receiving chamber 65 ( fig7 ) and functions to slidably receive a selected one of the stacked cover dispensing units 44 for movement between a first retracted position and a second dispensing position . base 63 is also provided with a longitudinal slot 69 that slidably receives the pusher segment 70 a of a finger engaging dispenser knob 70 that is adapted for movement in the direction of the arrow 73 of fig7 , between a first retracted position and a second dispensing position . when the finger engaging dispenser knob 70 is moved from its first retracted position to its second dispensing position , the pusher segment 70 a of a finger engaging dispenser knob engages a selected one of the cover dispensing units 44 , such as the device identified in fig1 by the numeral 44 x , and is moved within guide channel 67 from the inward position shown in fig1 to an outward position wherein it resides beneath the generally key - hole - shaped opening 64 . with the cover dispensing unit designated as 44 x , is in position beneath the generally key - hole - shaped opening 64 , the disposable cover assembly 56 can be interconnected with the stethoscope head in the manner illustrated in fig1 and 20 of the drawings . more particularly , in order to interconnect the disposable cover with the stethoscope head , the stethoscope head is moved from the position shown in fig1 into the downward position shown in fig2 where it is in engagement with the yieldably deformable central portion 58 a of the cover . a continued downward movement of the stethoscope head to the position shown in fig2 will cause it to move through the generally key - hole - shaped opening 64 and into the stethoscope covered position shown in fig2 and 21 of the drawings . as a stethoscope head moves downwardly through the keyhole shaped opening , the air - filled hollow tubes “ t ” of the stethoscope will pass through the segmented openings 53 and 54 a formed in the base 50 and in the rim 54 , respectively . importantly , as the stethoscope head moves downwardly into the position shown in fig2 and 21 of the drawings , the elastomeric bead 60 will release from the rim portion of the base and will move into engagement with the peripheral portion “ p ” of the stethoscope head , thereby removably securing the cover 56 to the stethoscope head . in this position , the flexible central membrane portion 58 a of the cover resides in close engagement with the lower surface “ s ” of the stethoscope head . once the cover is in position over the stethoscope head , the assemblage thus formed can be removed from the apparatus in the manner illustrated by the phantom lines in fig2 . following the examination of the patient , the cover assembly 56 can be conveniently removed from the stethoscope head and discarded into a suitable contaminated waste receptacle . in using this apparatus of the invention , the feed unit 46 is first mated with the base assembly 62 in the manner shown in fig1 of the drawings . this done , movement of the finger engaging dispenser knob 70 from its first retracted position into the position shown in fig1 of the drawings will cause the pusher segment 70 a of the finger engaging dispenser knob to engage the cover dispensing unit 44 x and move it forwardly within guide channel 67 . further movement of the finger engaging dispenser knob will cause the cover dispensing unit 44 x to move into the second outward position wherein it resides beneath the generally key - hole - shaped opening 64 ( see fig1 and 18 ). at the same time , the cover dispensing unit identified in fig1 and 18 by the numeral 44 y will drop downwardly into the position earlier occupied by cover dispensing unit 44 x . with the cover dispensing unit designated as 44 x thusly positioned beneath the generally key - hole - shaped opening 64 , the stethoscope head is moved from the position shown in fig1 into the downward position shown in fig1 where it is in engagement with the yieldably deformable central portion 58 a of the cover . a continued downward movement of the stethoscope head to the position shown in fig2 will cause it to move through the generally key - hole - shaped opening 64 and into the stethoscope covered position shown in fig2 and 21 of the drawings . as the stethoscope head moves downwardly through the keyhole shaped opening , the elastomeric bead 60 will release from the rim portion of the base and move into engagement with the peripheral portion “ p ” of the stethoscope head , thereby removably securing the cover 56 to the stethoscope head . once the cover is in position over the stethoscope head , the assemblage thus formed can be removed from the apparatus in the manner illustrated by the phantom lines in fig2 . following the examination of the patient , the cover assembly 56 can be conveniently removed from the stethoscope head and discarded into a suitable contaminated waste receptacle . to cover the stethoscope head with a fresh cover assembly 56 , finger engaging dispenser knob 70 is returned to its first starting position wherein the pusher segment 70 a is in engagement with the cover dispensing unit 44 y . forward movement of the finger engaging dispensing knob 70 from this starting position will cause the pusher segment 70 a of the finger engaging dispenser knob to move the cover dispensing unit 44 y forwardly within guide channel 67 into the second outward position wherein it resides beneath the generally key - hole - shaped opening 64 ( see fig1 and 18 ). at the same time , cover dispensing unit 44 y will urge cover dispensing unit 44 x outwardly of the base assembly . with a cover dispensing unit 44 y disposed beneath the keyhole shaped opening 64 , the fresh cover assembly 56 of cover dispensing unit 44 y can be removably affixed to the stethoscope head in the manner previously described . having now described the invention in detail in accordance with the requirements of the patent statutes , those skilled in this art will have no difficulty making changes and modifications in the individual parts or their relative assembly in order to meet specific requirements or conditions . such changes and modifications may be made without departing from the scope and spirit of the invention , as set forth in the following claims .

a protective cover for a stethoscope head and an apparatus for quickly and easily interconnecting the protective cover with the stethoscope head . the apparatus includes a cover - positioning device that positions the protective cover so that it can be quickly and easily interconnected with the stethoscope head . the cover positioning device includes a base having a planar portion and an upstanding rim connected to the planar portion , the upstanding rim defining an opening for receiving the stethoscope head therethrough . following examination of the patient using the stethoscope with the protective head covering , the protective covering can be quickly and easily removed from the stethoscope head and suitably disposed of so as to prevent skin bacteria and like contaminants that may have contaminated the protective cover during patient examination , from undesirably being transmitted to the next patient .

the inventor has made the novel discovery that mrna expression of constitutive androstane receptor ( car ; nr1i3 ) in the tissues of mammals is related to expression of genes involved in metabolic processes that influence metabolism of nutritional byproducts that are known to cause sensory problems of odor and taste in meat . specifically , the present discovery demonstrates that mrna expression of car is highly correlated with mrna expression of target genes important in fat metabolism as well as with genes involved in metabolism of endobiotics and xenobiotics . endobiotics refer to compounds that are byproducts of normal metabolism that may be toxic if not transformed and eliminated . xenobiotics refer to compounds derived from feed or other exogenous sources that also are potentially toxic to the animal . for example , the inventor has shown that increased expression of car mrna is related to increased expression of liver genes that control the metabolism of compounds that cause off - odors in pork . thus , one aspect of the present invention is to feed a supplement composed of one or more of the subject herbs , extracts , or vitamins such that car expression and / or activity is induced ( car inducers ) for the purpose of improving sensory qualities of odor and taste of meat . the subject supplement may be fed to male pigs for the purpose of reducing the presence of compounds in fat that cause sensory defects of odor and taste , a phenomenon known as “ boar taint ”. it is also expected that induction of car expression could be used to improve sensory qualities of meat from other genders of pigs , such as castrates and females . similarly , it is expected that induction of car could be used to improve sensory qualities of milk , as from a dairy cow . induction of car expression and activity may also be used to facilitate expression of enzymes involved in metabolism and excretion of potentially toxic foreign compounds ( xenobiotics ) as well as potentially toxic byproducts of metabolism ( endobiotics ) such as bilirubin . for example , car inducers may be used to mitigate the negative effects of toxic substances , such as mycotoxins on growth and reproductive traits in animals . it is known that particular plants , plant extracts , and vitamins affect metabolism by their influence on expression of nuclear receptors and nuclear receptor target genes in animal tissues , such as liver . one nuclear receptor in particular that is affected by these compounds is constitutive androstane receptor ( car ). artemisia spp ., tagetes lucida , and citrus peel contains scoparone , a compound known to induce car expression . in addition , alpha - tocopherol also is known to increase expression of car . gingko biloba has also been shown to increase expression of car target genes , such as those in the cyp2b family . two pilot studies have been conducted to demonstrate proof of concept for the use of artemisia capillaris , or other feed supplements containing the active component ( s ), to be fed to pigs for the reduction of boar taint compounds in pig fat . in the first study , entire male pigs ( non - breeding boars , 5 - 6 months of age ) were divided into three groups of fifteen boars each , comprising a control group and two treatment groups . the treatment groups were fed a dried aqueous extract of artemisia . capillaris at two different inclusion rates ( 15 g / day or 60 g / day ). after 8 days of treatment , pigs were transported and slaughtered at a plant by electrical stunning , and samples taken of kidney fat , back fat and liver . fat samples were placed in plastic bags and kept on ice before being trimmed and stored in vacuum bags at − 20 c . fat samples were analyzed according to previously validated methods ( dehnhard , m ., r . claus , m . hillenbrand , and a . herzog , 1993 : high performance liquid chromatographic ( hplc ) method for the determination of 3 - methylindole ( skatole ) and indole in adipose tissue of pigs . j . chromatogr . 616 , 205 - 209 ). the results showed that feeding of artemisia capillaris extract for a period of eight days prior to slaughter caused a reduction in the concentrations of skatole in back fat obtained from treated pigs . in the low - dose group , skatole levels were 50 % less than controls , and in the high dose group skatole levels were 60 % lower than controls . the reduction in skatole in both treatment groups compared with controls was statistically significant ( p & lt ; 0 . 05 ). a second pilot study was conducted in order to evaluate whether the aqueous extract used was more efficacious than feeding the dried artemisia capillaris . entire male pigs ( non - breeding boars , 5 - 6 months of age ; 15 per group ) were divided into a control and two treatment groups . the feed supplement treatments were either an aqueous extract of artemisia capillaris at an inclusion level of 30 g / day , or dried artemisia capillaris at an inclusion rate of 75 g / day . the results clearly indicated that feeding of the aqueous extract to boars for a period of eight days caused a reduction in the concentrations of skatole in the backfat at slaughter . fig1 : skatole concentrations in backfat of boars fed either aqueous extract or dried preparations of artemisia capillaris compared with controls . extract dried a . capillaris controls skatole ( μg / g fat ) 0 . 07 ± 0 . 01 * 0 . 11 ± 0 . 04 0 . 16 ± 0 . 03 * mean ± standard error ; extract versus controls , p & lt ; 0 . 02 . importantly , the percentage of boars that exhibited skatole concentrations in the backfat that were higher than the generally accepted sensory threshold (≧ 0 . 20 μg / g fat ) were reduced from 33 % in the controls , 13 . 3 % in the dried herb group , and to 0 . 0 % in the aqueous extract group . the reduction in the frequency of boars with high skatole levels in the extract group compared with controls was statistically significant ( p & lt ; 0 . 05 ; chi square analysis ). the use of the plant extracts and vitamins of the present invention for the reduction of off - odors and off - tastes associated with boar taint is novel and has not been reported or suggested in the prior art . artemisia capillaris has been suggested for use as a feed ingredient for improving growth rates of pigs ( pct wo 2006 / 006768 ), and was suggested to improve meat quality as measured solely by increasing fat content of meat . however the application for reducing boar taint and eliminating the need for castration of male pigs has not been indicated in the prior art . importantly , the present invention is novel in that the time period for feeding the compositions of the present invention are quite short , only from 2 to 21 days prior to slaughter . preferably the livestock feed of the present invention contains plant extracts obtained from artemisia species , such as artemisia capillaris , artemisia scoparia , artemisia annua , or from gingko biloba , and or peel or leaf extracts from citrus species . in a preferred embodiment , the plant extract includes an aqueous extract of artemisia species or , gingko biloba , or citrus peel . methods for preparation of aqueous plant extracts are well known in the prior art . it is also preferred to prepare a premix of plant extracts or vitamins , which may comprise an amount equivalent of 1 % to 80 % of such a premix . the premix may comprise other dietary supplements and may be mixed with conventional feed to obtain the feed of the invention , but this depends on the amount of premix to be used in the final feed composition . the amount of premix used is typically between 0 . 1 to 20 % based on the total weight of the mixed feed . for the purposes of reducing boar taint , the livestock feed of the present invention is intended to be fed for a short time period prior to slaughter , such as from 2 to 21 days . it is preferred to feed the supplement for 4 - 7 days prior to slaughter .

this invention relates to products and methods for improving sensory qualities of meat obtained from livestock . in particular , this invention relates to products and methods for reducing off - odors and off - tastes in meat from uncastrated male pigs or from other livestock fed on pasture . according to the invention , this is obtained by administering a product that contains an inducer of the constitutive androstane receptor . these include plant products derived from artemisia spp , citrus spp ., gingko biloba , tagetes lucida , as well as vitamin e . the car - inducer products of the invention may be used to reduce boar taint and thereby eliminate the need for castration of male pigs .

fig1 shows a footbath system 20 for livestock , including dairy animals such as 22 . a footbath pan 24 is provided for livestock to walk through . the pan has an inlet manifold 25 , fig1 , 4 , for receiving footbath liquid , and walls 26 , 28 , 30 , 32 containing the footbath liquid therein , for example as shown at 34 . a combined water containing and chemical mixing tank 36 , fig2 , 3 , is separate from pan 24 . tank 36 receives water from a water source 38 , and receives one or more chemicals from one or more chemical sources such as 40 for mixing in tank 36 to provide a pre - mixed footbath liquid . tank 36 supplies the pre - mixed footbath liquid to pan 24 , to be described . tank 36 , fig2 , has an inlet conduit 42 receiving water from water source 38 . tank 36 has one or more outlet conduits 44 , 46 , 48 , 50 supplying the pre - mixed footbath liquid to respective footbath pans such as 24 . tank 36 is disposed in serial fluid flow communication between inlet conduit 42 and outlet conduits 44 - 50 , such that water flows from upstream to downstream from inlet conduit 42 into tank 36 and then from tank 36 to outlet conduits 44 - 50 . tank 36 isolates outlet conduits 44 - 50 from inlet conduit 42 such that liquid pressure in outlet conduits 44 - 50 supplying pre - mixed footbath liquid to a respective pan such as 24 , e . g . at conduit 44 , fig4 , is isolated from water pressure in inlet conduit 42 supplying water from water source 38 . each of the outlet conduits 44 - 50 has a respective valve 52 , 54 , 56 , 58 , each having an on - state permitting liquid flow from tank 36 through the respective outlet conduit to the respective pan such as 24 , and having an off - state blocking liquid flow from tank 36 through the respective outlet conduit to the respective pan . a pump 60 , fig2 , 3 , pumps liquid from tank 36 via drain outlet conduit 62 then through outlet conduits 44 - 50 to a respective pan such as 24 at a pressure independent of water pressure from water source 38 . valves 52 - 58 are provided in respective outlet conduits 44 - 50 downstream of pump 60 . tank 36 has a chemical inlet conduit 64 , fig2 . chemical source 40 is a chemical container storing one or more chemicals and supplying the one or more chemicals through chemical conduit 64 to tank 36 . in one embodiment , container 40 is a hopper storing one or more powder chemicals , and chemical conduit 64 includes an auger transporting the powder chemicals to tank 36 . powder chemicals may be desirable in various applications for the convenience of the dairy farmer enabling him to merely dump the powder into hopper 40 for storage and subsequent usage . the chemicals typically include , but are not limited to , germicides , bacteriacides , other medical treatments , and so on , to treat the hooves of livestock . tank 36 has the noted outlet conduits 62 , 44 - 50 , fig2 , 3 , supplying the pre - mixed footbath liquid to one or more footbath pans such as 24 , fig4 , 24 a , fig5 , etc . the outlet conduit includes an upstream segment 62 receiving the pre - mixed footbath liquid from tank 36 , and a plurality of parallel downstream segments 44 - 50 receiving the pre - mixed footbath liquid in parallel from upstream segment 62 and supplying the pre - mixed footbath liquid to respective pans such as 24 , 24 a , and so on . valves 52 - 58 are provided in respective downstream segments 44 - 50 of the outlet conduit . in the preferred embodiment , footbath pan 24 is axially elongated along a longitudinal axis 70 , fig4 , 6 . the noted walls include upstream and downstream end walls 26 and 30 , and a pair of sidewalls 28 and 32 extending axially longitudinally therebetween . end walls 26 , 30 and sidewalls 28 , 32 have a height sufficient to contain footbath liquid around the livestock &# 39 ; s hooves . the livestock initially steps into the pan over upstream end wall 26 and then walks axially ( rightwardly in fig1 , 4 , 6 , 7 ) while between sidewalls 28 , 32 and then exits the pan by stepping over downstream end wall 30 . each of end walls 26 and 30 has a respective length extending laterally along lateral direction 72 between sidewalls 28 and 32 . each of sidewalls 28 , 32 has a respective length extending longitudinally along longitudinal axial direction 70 between end walls 26 and 30 . the above noted outlet conduit 44 from tank 36 is connected to a pan inlet 74 supplying liquid into pan 24 at manifold 25 . a door 76 , fig4 - 8 , has a closed position , fig4 , retaining liquid in pan 24 , and has an open position , fig5 , 7 , 8 , draining liquid from the pan , as shown at arrow 78 . the door forms at least a portion of , and preferably most or all of , a designated one of the noted walls 26 - 32 , preferably downstream end wall 30 . door 76 preferably has a length of at least 50 % of the length of the noted designated wall , e . g . downstream end wall 30 , for reasons noted below . door 76 has an upper edge 80 pivoted on a hinge 82 about an upper pivot axis , and has a lower edge 84 swingable in an arc 86 about the noted upper pivot axis between the closed position and the open position . door 76 is preferably at the downstream end wall and extends laterally along lateral direction 72 substantially the entire lateral length of downstream end wall 30 , which in the preferred embodiment enhances desired flow , noted below . door 76 is actuated between the closed and open positions by pneumatic cylinder 88 . pan inlet 74 , fig4 , preferably supplies liquid in non - turbulent flow into and along pan 24 via inlet manifold 25 . inlet manifold 25 has one or more flow ports 90 , fig4 , provided by one or more slots or openings or the like , along a given lateral span 91 and preferably discharging liquid at high volume , low velocity flow , namely selected to provide a reynolds number less than 600 , 000 , to provide non - turbulence . in a further preferred embodiment , the flow is selected to provide a reynolds number between 300 , 000 and 600 , 000 , to provide non - turbulent transitional flow . in a yet further preferred embodiment , the flow is selected to provide a reynolds number less than 300 , 000 , to provide non - turbulent laminar sheet flow . door 76 is distally opposite flow ports 90 and preferably has a length at least as great as the noted lateral span 91 thereof . in the preferred embodiment , the noted non - turbulent flow is along a rectilinear flow path from the inlet at flow ports 90 to the outlet at door 76 without eddy currents , and further preferably in the noted laminar sheet flow . at the downstream end , if the lateral length of door 76 is not as great as the lateral length of downstream end wall 30 , then it is preferred that tapered ramp surfaces be provided as shown at 92 , 94 , to guide the noted flow in non - turbulent manner , and without eddy currents , to door 76 . the present system provides a method for treating hooves of livestock , including dairy animals . the method includes the steps of providing a footbath system including a footbath pan 24 for livestock to walk through , the pan having walls 26 - 32 containing footbath liquid therein , providing a combined water containing and chemical mixing tank 36 separate from pan 24 , supplying water from a water source to the tank , supplying one or more chemicals from a chemical source 40 to the tank , mixing the water and the one or more chemicals in the tank to provide a pre - mixed footbath liquid , and supplying the pre - mixed footbath liquid from the tank to the pan . the method includes providing the tank with an inlet conduit 42 receiving water from the water source 38 , providing the tank with an outlet conduit 62 , 44 - 50 , supplying the pre - mixed footbath liquid to one or more pans 24 , 24 a , etc ., disposing the tank 36 in serial fluid flow communication between inlet conduit 42 and outlet conduit 62 , 44 - 50 , supplying water to flow from upstream to downstream from inlet conduit 42 into tank 36 and then from tank 36 to outlet conduit 62 , 44 - 50 , isolating outlet conduit 62 , 44 - 50 from inlet conduit 42 by tank 36 therebetween such that liquid pressure in outlet conduit 62 , 44 - 50 supplying the pre - mixed footbath liquid to pans 24 , 24 a , etc ., is isolated from water pressure in inlet conduit 42 from water source 38 . the method further includes providing tank 36 with a chemical inlet conduit 64 , providing the chemical source 40 as a chemical container storing one or more chemicals , and supplying the one or more chemicals from the container 40 through the chemical conduit 64 to tank 36 . the method further includes providing the container 40 as a hopper , storing one or more powder chemicals in the hopper , providing the chemical conduit 64 as an auger , and transporting powder chemicals with the auger to tank 36 . the method further includes providing the pan with a door 76 having a closed position retaining liquid in the pan , and having an open position draining liquid from the pan , providing the pan with an inlet 74 including inlet manifold 25 , and supplying liquid from the inlet manifold 25 at flow ports 90 in non - turbulent flow into and along pan 24 . the method includes supplying the liquid in non - turbulent flow into and along pan 24 in each of the noted closed and open positions of door 76 . in one embodiment , the system has a drain mode , a flush mode , and a fill mode . in the drain mode , the method preferably includes opening door 76 without liquid flow into pan 24 at inlet 74 . in the flush mode , the method preferably includes opening door 76 and supplying liquid in non - turbulent flow from the inlet at flow ports 90 into and along pan 24 . in the fill mode , the method preferably includes closing door 76 and supplying the liquid at inlet 74 in non - turbulent flow into and along pan 24 . in the fill mode , the method further preferably includes additionally supplying one or more chemicals into pan 24 through the same inlet 74 and same manifold 25 and same flow ports 90 supplying water into the pan in non - turbulent flow . the present system desirably eliminates high velocity jet nozzle flow into the pan , and consequent turbulence and eddy currents . prior art systems typically include an agitation phase prior to the drain phase , wherein high velocity turbulent and eddy current flow is used for agitation , followed by draining and flushing . the present system desirably eliminates turbulent agitation and eddy current flow because of undesirable splatter and jet spray , and undesirable release of bacteria and odor upon break - up and / or dissolution of manure and the like . laterally elongated door 76 is desired over prior smaller discharge orifices because door 76 facilitates easy drainage without agitation and turbulence . the high volume , low velocity inlet flow at ports 90 at reynolds number less than 600 , 000 , and preferably less than 300 , 000 to provide laminar sheet flow , is further desired because it enables the noted chemicals to be introduced through the same inlet flow ports 90 as the water , without requiring a second separate set of one or more chemical inlets as in the prior art using a first set of high velocity jet nozzle ports for water inlet , and a second set of ports for chemical inlet . in another embodiment , one or more liquid chemical containers 102 , 104 , 106 , fig2 , may be used instead of , or in addition to , powder chemical container 40 . the liquid chemicals are pumped by respective pumps 108 , 110 , 112 through respective conduits 114 and 116 , 118 and 120 , 122 and 124 , from respective storage tank containers 102 , 104 , 106 to mixing tank 36 . the footbath liquid may include water plus one or more chemicals , or water only , or one or more chemicals only . the system may be manually controlled , or in another embodiment may be automated including a control panel 126 or the like responsive to livestock count , sensed chemical conditions in the footbaths , timing patterns , including time of day or week , and so on . in a further embodiment , one or more of the footbaths may have folding hatch doors such as 128 , 130 , fig5 , for closing and covering the footbath when not in use . in further embodiments , auger 64 may instead be a conveyor or some other transport mechanism transporting chemicals therealong to tank 36 . in further embodiments , the various chemical inlets may be unused or not connected , e . g . for a water - only flush , fill , etc ., wherein tank 36 only contains water , which water is the sole constituent of the footbath liquid . in the foregoing description , certain terms have been used for brevity , clearness , and understanding . no unnecessary limitations are to be implied therefrom beyond the requirement of the prior art because such terms are used for descriptive purposes and are intended to be broadly construed . the different configurations , systems , and method steps described herein may be used alone or in combination with other configurations , systems and method steps . it is to be expected that various equivalents , alternatives and modifications are possible within the scope of the appended claims .

a footbath system for livestock includes a water and / or chemical containment tank , a footbath pan with a drainage exit door , non - turbulent flow , and a multiple branch system .

before describing in detail the particular game or toy in accordance with the present invention , it should be observed that the present invention resides primarily in a novel and non - obvious combination of hardware components . accordingly , the hardware components have been represented by conventional elements in the figures , showing only those specific details that are pertinent to the present invention so as not to obscure the disclosure with structural details that would be readily apparent to those skilled in the art having the benefit of the description herein . [ 0008 ] fig1 illustrates a game 10 including a base 12 . a plunger 16 and a rotatable handle 18 extend above the base 12 for manipulation by the player . also affixed to the base 12 is a support member 20 for supporting a pitching mechanism 22 . the support member 20 and the pitching mechanism 22 are joined by a hinge 24 that allows the pitching mechanism 22 to be rotated relative to the support member 20 . affixed to the pitching mechanism 22 is a hollow cup or container 26 . further affixed to the pitching mechanism 22 is a hook 28 . a spring member 30 is affixed near the top edge of the support member 20 as shown . further affixed to the base is a hook 32 . as a first step in operating the game , the pitching mechanism 22 is rotated away from the base 12 as shown by the phantom representation in fig1 . this movement causes flexing of the spring member 30 as also shown by the phantom representation in fig1 . after the pitching mechanism 22 is rotated into this position , the hook 28 is engaged to the hook 32 so as to hold the pitching mechanism biased against the spring member 30 . in a second step of the game , the player chooses a random number that can range from one to about 20 or 30 . in fact , any number can be chosen , but to make the game manageable the maximum chosen number should be in the range of 20 to 30 . the player then rotates the rotatable handle 18 through a number of turns equal to the chosen random number . turning to fig2 it is seen that the rotatable handle 18 is affixed to a disk 38 , and rotates within a well 42 of a bottom frame member 34 . a top frame member 33 and a bottom frame member 34 of the base 12 are connected at the ends thereof by vertical members 35 and 36 . rotation of the rotatable handle 18 turns the disk 38 . the disk 38 includes a plurality of notches 42 located around the circumference thereof . see fig3 . returning to fig2 there is shown an arm 44 extending from a support member 46 , which is attached to the top frame member 33 . the arm 44 , which in one embodiment is a relatively thin metallic piece , sequentially engages the notches 42 as the disk 38 is rotated by the rotatable handle 18 , which creates a detent mechanism . as the player rotates the rotatable handle 18 , each notch is in turn engaged by the arm 44 , so that the game player simply counts the number of detent actions until the chosen random number is reached . note further from fig3 that the disk 38 includes a plurality of randomly spaced holes 50 . as the disk 38 is rotated , one of these holes may be rotated into position beneath the plunger 16 . the plunger 16 is biased upwardly by a helical spring 52 , but can be pushed downwardly by the player . after the random number is entered by rotating the rotatable handle 18 , the player pushes downward on the plunger 16 . if none of the holes 50 is aligned with the plunger 16 , the plunger will simply strike the top surface of the disk 38 and return to its upward position through operation of the helical spring 52 . alternatively , if one of the holes 50 has been rotated into position below the plunger 16 , the downward force on the plunger 16 , extends the plunger through the hole 50 , where it contacts a lever member 54 . see fig2 . as shown , the lever member 54 is attached at the fulcrum end to a u - shaped spring steel member 58 , which biases the lever member 54 into the position shown . in one embodiment , the rotatable handle 18 passes through the lever member 54 . in another embodiment a groove or notch can be cut into the lever member 54 , allowing sufficient clearance for the rotatable handle 18 . when the plunger 16 is depressed through a hole 50 into contact the lever member 54 , the lever member 54 rotates against the bias provided by the spring steel member 58 . this movement of the lever member 54 causes movement of the hook 32 as shown by the arrow in fig2 . as the hook 32 rotates about its attachment point with the support member 20 , it disengages from the hook 28 attached to the pitching mechanism 22 . at this point , the bias provided by the spring member 30 drives the pitching mechanism 22 into its equilibrium position , at which point water or another substance located within the cup or container 26 is accelerated out of the cup . typically , during the operation of rotating the rotatable handle 18 and depressing the plunger 16 , the player will be situated in front of the game 10 and therefore , in line with the material accelerated out from the cup or container 26 . thus the selection of the right random number may result in the player getting wet and thereby being declared a loser . as can be appreciate by those skilled in the art , several different embodiments are possible for the present invention . for instance , the spring member 30 and the spring steel member 58 can be replaced by hinge and spring mechanisms to provide an equilibrium position for the associated members while also serving to apply forces to move the associated members as required . while the invention has been described with reference to a preferred embodiment , it will be understood by those skilled in the art that various changes may be made and equivalent elements may be substituted for elements thereof without departing from the scope of the present invention . in addition , modifications may be made to adapt a particular situation more material to the teachings of the invention without departing from the essential scope thereof . therefore , it is intended that the invention not be limited to the particular embodiment disclosed as the best mode contemplated for carrying out this invention , but that the invention will include all embodiments falling within the scope of the appended claims .

a game or toy device into which the player enters a random number is disclosed . the random number is entered by rotating a handle after which a plunger is depressed . entry of the random number may permit free depression of the plunger and thereby allow the plunger to activate a pitching mechanism for ejecting a substance in the direction of the game player .

the use of sub - units ( a chain and b chain of the ricin toxin ) was attempted to address concerns about the safety and efficacy of the prior art vaccines to protect from pathological effects of exposure to ricin toxin . since the sub - units are far less toxic than ricin ( 2 - 6 logs difference in toxicity ) it was thought that use of subunits would provide reliable protection against the ricin toxin without exposure to untoward side effects . the problems encountered in use of sub - units varied . it was found that the ricin a chain was very sensitive to treatment with formalin . when the ricin a chain is treated with formalin , it is possible to inactivate toxic properties by use of formalin alone without heating . it is possible to protect susceptible mammals with either the a or b chain . vaccination by ricin b chain will protect from systemic challenge , but will not protect from aerosol challenge with the ricin whole toxin . when the animals vaccinated with only the b chain survive ( infrequently ) aerosol challenge there is always measurable titer to ricin a chain in their sera . when ricin a chain ( rta ) is used to immunize , protection against both systemic and aerosol challenge is effected . hence , the immunization using ricin a chain is a preferred form of protection against pathological effects of ricin toxin . during preliminary studies , administrations of ricin b chain and ricin a chain were studied . untreated ricin chains a and b were introduced separately into mice , with the a chain being administered at a dosage of 5 . 0 μg / mouse / injection and the b chain administered at dosage of 3 . 0 μg / mouse / injection . the time between administration of the separate subunits varied from 24 hour to 7 days . the protective response was assessed by exposing the mice to challenge with lethal ricin aerosol . the results are shown below : ______________________________________time between dosingwith a chain and b chain survival rate______________________________________24 hrs . 6 / 748 hrs . 5 / 57 days 4 / 4controls : 0 / 8______________________________________ a second test employed the administration of b chain followed in two days by administration of a chain . the administration of the dosages with the two day separation between the chains was repeated with one week between the first administration of the a chain and the second administration of the b chain followed by a third similar dosing . twelve days after the administration of the last dose of a chain the mice were challenged with aerosol ricin . all twelve animals survived . the two component chains of the ricin toxin may be purchased separately (≧ 99 % pure ). systemically each of the chains can be shown to be at least two logs less toxic than the native toxin . when both untreated chains were introduced simultaneously death resulted . co - introduction of one treated and one untreated chain or two treated chains leads to active immunity . when the untreated a and b chains are introduced separately with at least a 24 hour interval between introduction of the second chain , active immunity resulted without lethal results . the order of introduction of the chains is not important . when there was co - introduction of one treated and one untreated chain or of two chains wherein both chains had been treated with formalin the animals developed effective immunity without lethal effects . it is suggested that timed release using biodegradable polymer could be administered together if the system avoids administration of both a chain and b chain in such close proximity that both would be present in the host at the same time . ricin a chain only was used to immunize mice with dosages of 5 . 0 μg / mouse / injection and 3 . 0 μg / mouse / injection being administered . repeat dosages were administered two weeks apart . ______________________________________dosage number of doses results______________________________________5 . 0 μg 2 1 / 5control 0 / 83 . 0 μg 3 5 / 12control 0 / 10______________________________________ when the immunizations were administered three times at two week intervals at dosage of 5 . 0 μg / mouse / injection , all mice in four independent tests survived challenge with aerosol administration of ricin . hence , it is seen that with administration of only ricin a chain ( rta ) it is possible to provide effective immunization against the lethal effects of aerosol administered ricin without use of the b chain as an antigen . it is also seen that the survival is related to dosage amount and number of doses administered . the antigenic agents of the invention may be administered by any means that will result in the active agent contacting tissue capable of immune response . administration may be , for example , by injection ( for example , subcutaneous , intradermal , or intramuscular ), by contact with the nasopharyngeal mucosa , or by application to abraded dermal tissue . the compositions for administration may contain adjuvants such as alum . the administration of rta alone or preceded by or followed by administration of the b chain or administration of the treated , separate chains to larger animals or to cells in vitro can also be used to elicit antibodies for administration to non - protected patients who have been exposed to the ricin toxin . because the chains administered separately are far less toxic , the living systems used to provide the antibodies are not adversely affected thereby . the sub - units used for protection by methods of the invention can also be produced in culture by recombinant means known in the art . eukaryotic cells , e . coli and yeast are suggested cells use in making recombinant rta . a example of such production is seen in u . s . pat . no . 4 , 689 , 401 to ferris , which is incorporated herein by reference . equivalents of the native rta should contain the fragment which interacts with the 60s ribosome .

it is now possible to immunize susceptible mammals against the pathological effect of exposure to rice , including inhalation of ricin , by administration of an immunogenic effective amount of ricin toxin sub - units , including subunits of both the a chain and the b chain of the ricin toxin given separately to provide safe , efficacious protection .

key targets in a strategy to retard cancer cell proliferation are the prenylation enzymes , such as the enzyme ftase . by reducing or destroying ftase activity , we have found that we can prevent ras farnesylation , which in turn prevents localization of ras at the inner surface of the cancer cell membrane , thereby preventing ras from causing the cancer cells to divide and proliferate . many substances are known to block ftase activity and prevent farnesylation of ras . they consist of inhibitors of the enzyme ftase , and they generally operate by blocking the binding of ras or fpp ( or both ) to ftase . without the normal substrates ( ras and fpp ) being able to bind to ftase , ftase cannot transfer the farnesyl group from fpp to ras . inhibitors structurally mimic one or both of the natural substrates of the enzyme , ras and / or fpp . their binding is noncovalent and reversible . in other words , the binding of the inhibitor to ftase does not involve the formation of covalent bonds between the inhibitor and ftase . instead , hydrophobic forces , hydrogen bonding , electrostatic attraction , etc . are the forces principally responsible for binding of the inhibitor to the enzyme ftase . these binding forces allow the inhibitor to block the site on ftase where the normal substrates need to bind for farnesylation of ras to take place . we have found that a potentially more useful method of preventing ftase from farnesylating ras can be achieved , namely inactivation of ftase . interaction of ftase with substances that are inactivators results in the covalent attachment of a part or all of the inactivator to ftase . in principle , the covalent attachment can be irreversible or nearly irreversible . also , the specificity of the inhibitor for the target enzyme ftase can be more finely tuned because it is based on two parameters : ( 1 ) structural similarity to the normal substrates ( fpp and / or ras ); ( 2 ) a reactivity that is appropriate for the chemical groups that are a part of the enzyme ftase , particularly those that participate in the actual process catalyzed by ftase ( i . e ., those that more - or - less directly participate in the transfer of the farnesyl group of fpp to ras ). the fact that fpp loses pyrophosphate , which is anionic , might be indicative of the presence of a structural feature in ftase that would assist in the displacement of pyrophosphate , for example by ionic bonding to a positively charged group in the enzyme &# 39 ; s active site . one such possibility is the side chain of arginine [ r — nh ═ c ( nh 2 ) 2 + ]. indeed , the x - ray crystallographic structure determination of ftase reveals the presence of arginine residues in the vicinity of the hypothetical binding site of fpp , which further suggests the possible role of arginine in assisting with the removal of pyrophosphate as the farnesyl group becomes bonded to ras . some chemical reagents that react with the side chain of arginine residues in proteins possess a functional group that comprises at least two adjacent carbonyl groups , i . e ., o ═ c — c ═ o . such a functional group binds covalently to the nitrogens of the side chain of arginine , causing the vicinity of the arginine to become blocked and possibly the positive charge to be lost . hypothetical structures of the adduct ( s ) of the arginine side chain and the dicarbonyl compound phenylglyoxal are shown below : our invention consists of chemical agents bearing at least two adjacent carbonyl groups ( o ═ c — c ═ o ) designed to react ( i . e ., covalently interact ) with the arginine side chain . furthermore , to target better the substance to ftase in preference to other enzymes , and to target better the substance to the active site of ftase in preference to the external or noncatalytic portion of the enzyme , a preferred embodiment of the invention consists of a substance with at least one dicarbonyl functional group and other structural features which favor the binding of the agent to the active site of ftase . for example , the active site of ftase is hydrophobic to bind the natural substrate fpp which contains a hydrophobic region ( i . e ., the farnesyl group ). a hydrophobic substance that also contains the dicarbonyl functional group is preferred , a class represented by the following generalized structural fomula : wherein r and r ′ represent hydrogen and / or a hydrophobic group , such as an alkyl , alkenyl , alkynyl , and aryl group . the group can be cyclic or noncyclic , branched or unbranched , with or without unsaturation ( e . g ., aryl , alkenyl ) or substituents . preferably the group is designed to impart specificity of the substance for binding to and / or inactivation of ftase . the term “ alkyl ,” as used alone or in combination herein , refers to an unsubstituted or optionally substituted , straight , or branched chain saturated hydrocarbon group containing from one to twenty - five carbon atoms , preferably from one to fifteen carbons , such as methyl , ethyl , n - propyl , n - butyl , pentyl , hexyl , heptyl , octyl , the various branch chain isomers thereof , such as isopropyl , isobutyl , sec - butyl , tert - butyl , isohexyl and the like . the alkyl group may be optionally substituted by one or more substituents , and generally no more than three , and most often just one substituent . preferred optional substituents include halo , alkoxy , amino , mono - and di - substituted amino , aryl , carboxylic acid , heterocyclo , heteroaryl , cycloalkyl , hydroxy , trifluoromethoxy and the like . the term “ lower alkyl ” refers to such alkyl groups containing from one to five carbon atoms . the term “ alkoxy ,” as used alone or in combination herein , refers to an alkyl group , as defined above , covalently bonded to the parent molecule through an — o - linkage , such as methoxy , ethoxy , propoxy , isopropoxy , butoxy , t - butoxy and the like . the term “ alkoxyalkyl ” refers specifically to an alkyl group substituted with an alkoxy group . the term “ aryloxy ,” as used alone or in combination herein , refers to an aryl group , as defined below , covalently bonded to the parent molecule through an — o - linkage . an example of an aryloxy is phenoxy . the term “ cycloalkoxy ,” as used alone or in combination herein , refers to a cycloalkyl group , as defined below , covalently bonded to the parent molecule through an — o - linkage . the term “ alkylthio ,” as used alone or in combination herein , refers to an alkyl group , as defined above , covalently bonded to the parent molecule through an — s - linkage . the term “ alkenyl ,” as used alone or in combination herein , refers to an alkyl group , as defined above , containing one or more carbon - carbon double bonds , preferably one or two double bonds . examples of alkenyl include ethenyl , propenyl , 1 , 3 - butadienyl , and 1 , 3 , 5 - hexatrienyl . the term “ alkynyl ,” as used alone or in combination herein , refers to an alkyl group , as defined above , containing one or more carbon - carbon triple bonds , preferably one or two triple bonds . the term “ cycloalkyl ,” as used alone or in combination herein , refers to an unsubstituted or optionally substituted , saturated cyclic hydrocarbon group containing three to eight carbon atoms . the cycloalkyl group may optionally be substituted by one or more substituents , and generally no more than three , and most often just one substituent . preferred optional substituents include alkyl , halo , amino , mono - and di - substituted amino , aryl , hydroxy and the like . the term “ haloalkyl ” is a species of alkyl as defined herein , and particularly refers to an alkyl , preferably a lower alkyl , substituted with one or more halogen atoms , and preferably is a c 1 to c 4 alkyl substituted with one to three halogen atoms . one example of a haloalkyl is trifluoromethyl . the term “ alkanoyl ” as used alone or in combination herein refers to an acyl radical derived from an alkanecarboxylic acid ( alkyl - c ( o )—), particularly a lower alkanecarboxylic acid , and includes such examples as acetyl , propionyl , butyryl , valeryl , and 4 - methylvaleryl . the term “ aroyl ” means an acyl radical derived from an aromatic carboxylic acid , such as optionally substituted benzoic or naphthoic acids and specifically including benzoyl and 1 - naphthoyl . the term “ aminocarbonyl ” means an amino - substituted carbonyl ( carbamoyl or carboxamide ) wherein the amino group is a primary amino (— nh 2 ). substituted aminocarbonyl refers to secondary ( mono - substituted amino ) or tertiary amino ( di - substituted amino ) group , as defined below , preferably having as a substituent ( s ) a lower alkyl group . the term “ aminoalkanoyl ” means an amino - substituted alkanoyl wherein the amino group is a primary amino group (- alkyl - c ( o )— nh 2 ). the term “ substituted aminoalkanoyl ” refers to related secondary ( mono - substituted amino ) or tertiary amino ( di - substituted amino ) group , as defined below . the term “ carbocycloalkyl ” when used alone or in combination refers to an unsubstituted or optionally substituted , stable , saturated or partially unsaturated monocyclic , bridged monocyclic , bicyclic , and spiro ring carbocycles of 3 to 15 carbon atoms such as cyclopropyl , cyclobutyl , cyclopentyl , cyclohexyl , cycloheptyl , bicyclohexyl , bicyclooctyl , bicyclononyl , spirononyl and spirodecyl . cycloalkyls are thus one specific subset of carbocycloalkyls . the term “ optionally substituted ” as it refers to “ carbocycloalkyl ” herein indicates that the carbocycloalkyl group may be substituted at one or more substitutable ring positions by one or more groups independently selected from alkyl ( preferably lower alkyl ), alkoxy ( preferably lower alkoxy ), nitro , monoalkylamino ( preferably a lower alkylamino ), dialkylamino ( preferably a di [ lower ] alkylamino ), cyano , halo , haloalkyl ( preferably trifluoromethyl ), alkanoyl , aminocarbonyl , monoalkylaminocarbonyl , dialkylaminocarbonyl , alkylamido ( preferably lower alkylamido ), alkoxyalkyl ( preferably a lower alkoxy [ lower ] alkyl ), alkoxycarbonyl ( preferably a lower alkoxycarbonyl ), alkylcarbonyloxy ( preferably a lower alkylcarbonyloxy ) and aryl ( preferably phenyl ), said aryl being optionally substituted by halo , lower alkyl and lower alkoxy groups . generally , there is no more than one optional substituent . the term “ heterocyclo ” as used , alone or in combination , herein refers to an unsubstituted or optionally substituted , stable , saturated , or partially unsaturated , monocyclic , bridged monocyclic , bicyclic , and spiro ring system containing carbon atoms and other atoms selected from nitrogen , sulfur and / or oxygen . preferably , a heterocyclo group is a 5 or 6 - membered monocyclic ring or an 8 - 11 membered bicyclic ring which consists of carbon atoms and contains one , two , or three heteroatoms selected from nitrogen , oxygen and / or sulfur . heterocyclo includes benz - fused monocyclic cycloalkyl groups having at least one such heteroatom . the term “ optionally substituted ” as it refers to “ heterocyclo ” herein indicates that the heterocyclo group may be substituted at one or more substitutable ring positions by one or more groups independently selected from alkyl ( preferably lower alkyl and including haloalkyl ( preferably trifluoromethyl )), alkoxy ( preferably lower alkoxy ), nitro , monoalkylamino ( preferably a lower alkylamino ), dialkylamino ( preferably a di [ lower ] alkylamino ), cyano , halo , alkanoyl , aminocarbonyl , monoalkylaminocarbonyl , dialkylaminocarbonyl , alkylamido ( preferably lower alkylamido ), alkoxyalkyl ( preferably a lower alkoxy [ lower ] alkyl ), alkoxycarbonyl ( preferably a lower alkoxycarbonyl ), alkylcarbonyloxy ( preferably a lower alkylcarbonyloxy ) and aryl ( preferably phenyl ), said aryl being optionally substituted by halo , lower alkyl and lower alkoxy groups . generally , there is no more than one optional substituent . several non - limiting examples of such heterocyclo groups are illustrated below : the heterocyclo group may be , and generally is attached to the parent structure through a carbon atom , or alternatively may be attached through any heteroatom of the heterocyclo group that results in a stable structure . the term “ heteroaryl ” as used alone or in combination , herein refers to an unsubstituted or optionally substituted , stable , aromatic monocyclic or bicyclic ring system containing carbon atoms and other atoms selected from nitrogen , sulfur and / or oxygen . preferably , a heteroaryl group is a 5 - or 6 - membered monocyclic ring ( optionally benzofused ) or an 8 - 11 membered bicyclic ring which consists of carbon atoms and contains one , two , or three heteroatoms selected from nitrogen , oxygen and / or sulfur . the term “ optionally substituted ” as it refers to “ heteroaryl ” herein indicates that the heteroaryl group may be substituted at one or more substitutable ring positions by one or more groups independently selected from alkyl ( preferably lower alkyl and including haloalkyl ( preferably trifluoromethyl )), alkoxy ( preferably lower alkoxy ), nitro , monoalkylamino ( preferably a lower alkylamino ), dialkylamino ( preferably a di [ lower ] alkylamino , cyano , halo , alkanoyl , aminocarbonyl , monoalkylaminocarbonyl , dialkylaminocarbonyl , alkylamido ( preferably lower alkylamido ), alkoxyalkyl ( preferably a lower alkoxy [ lower ] alkyl ), alkoxycarbonyl ( preferably a lower alkoxycarbonyl ), alkylcarbonyloxy ( preferably a lower alkylcarbonyloxy ) and aryl ( preferably phenyl ), said aryl being optionally substituted by halo , lower alkyl and lower alkoxy groups . generally , there is no more than one optional substituent . several non - limiting examples of such heteroaryl groups are illustrated below : the heteroaryl group may be , and generally is attached to the parent structure through a carbon atom or alternatively may be attached through any heteroatom of the heteroaryl group that results in a stable structure . in the foregoing structures it also is contemplated that a nitrogen could be replaced with an n - oxide . both heterocyclo and heteroaryl also are intended to embrace benzo fused structures such as 1 , 2 - methylenedioxybenzene and 1 , 4 - benzodioxan . the terms “ halo ” and “ halogen ” as used herein to identify substituent moieties , represent fluorine , chlorine , bromine or iodine , preferably chlorine or fluorine . the term “ aryl ,” when used alone or in combination , refers to an unsubstituted or optionally substituted monocyclic or bicyclic aromatic hydrocarbon ring system having 6 to 12 ring carbon atoms . preferred are optionally substituted phenyl , 1 - naphthyl , or 2 - naphthyl groups . the aryl group may optionally be substituted at one or more substitutable ring positions ( generally at no more than three positions and most often at one or two positions ) by one or more groups independently selected from alkyl ( including haloalkyl ( preferably trifluoromethyl and difluoromethyl )), alkenyl , alkynyl , alkoxy , aryloxy , nitro , hydroxy , amino , mono - and di - substituted amino , cyano , halo , alkanoyl , aminocarbonyl , carboxylic acid , carboxylic acid esters , carboxylic acid amide , an optionally substituted phenyl ( optionally substituted by halo , lower alkyl and lower alkoxy groups ), heterocyclo , or heteroaryl . preferably , the aryl group is phenyl optionally substituted with up to four and more usually with one or two groups , preferably selected from lower alkyl , lower alkoxy , as well as cyano , trifluoromethyl and halo . the terms “ aralkyl ” and “( aryl ) alkyl ,” alone or in combination are a species of alkyl as defined herein , and particularly refers to an alkyl group as defined above in which one hydrogen atom is replaced by an aryl group as defined above , and includes benzyl , and 2 - phenylethyl . the terms “( heterocyclo ) alkyl ” and “( heteroaryl ) alkyl ” alone or in combination can be considered a species of alkyl as defined herein , and particularly refers to an to an alkyl group as defined above in which one hydrogen atom is replaced by a heterocyclo group as defined above , or by a heteroaryl group as defined above . the term “ alkoxycarbonyl ” alone or in combination means a radical of the formula — c ( o )- alkoxy , in which alkoxy is as defined above . the term “ alkylcarbonyloxy ” alone or in combination means a radical of the formula — o — c ( o )- alkyl , in which alkyl is as defined above . the term “ alkoxyalkanoyl ” alone or in combination means a radical of the formula — alkyl - c ( o )— o - alkyl . the term “ carboxyalkyl ” alone or in combination means a radical of the formula — alkyl - c ( o )— oh . the term “ substituted amino ” embraces both mono and di - substituted amino . these terms , alone , or in combination , mean a radical of the formula — nr 1 r ″, where , in the case of mono - substitution , one of r ′ and r ″ is a hydrogen and the other is selected from alkyl , cycloalkyl , aryl , heterocyclo , ( aryl ) alkyl , ( heterocyclo ) alkyl , heteroaryl and hetero ( aryl ) alkyl ; in the case of di - substitution , r ′ and r ″ are independently selected from alkyl , cycloallyl , aryl , heterocyclo , and heteroaryl , or r ′ and r ″ together with the nitrogen atom to which they are both attached form a three to eight - membered heterocyclo or heteroaryl radical . the term “ amido ” refers to the group (— nh —) and the term “ substituted amido ” embraces a radical of the formula (— nr —) where r ′ has the meaning above in connection with substituted amino . the terms “ alkanoylamido ,” “ aroylamido ,” “ heterocyclocarbonylamido ” and “ heteroaroylamido ” mean groups of the formula r — c ( o )— nh — where r is an alkyl , aryl , heteroaryl or heterocyclo group . the terms “ heteroaroyl ” and “ heterocyclocarbonyl ” when used alone or in combination means groups of the formula r — c ( o )— where r is a heteroaryl or heterocyclo group . unless otherwise defined , the term “ optionally substituted ” as used herein , refers to the substitution of a ring system at one or more positions with one or more groups selected from : c 1 - 5 alkyl , c 1 - 5 alkoxy , an optionally substituted phenyl , cyano , halo , trifluoromethyl , c 1 - 5 alkoxycarbonyl , c 1 - 5 alkyl carbonyloxy , mono - and bis -( c 1 - 5 alkyl )- carboxamide , c 1 - 5 alkylamido , nitro , and mono - and bis -( c 1 - 5 alkyl )- amino . applicants recognize that there may be some overlap in some of the definitions of the various radical groups . specific groups are mentioned , however , such as ( aryl ) alkyl , and may be particularly identified in the claims , in order to emphasize their positive inclusion in the described subject matter , as not only an optional substituent . as used herein , when a particular radical generally understood to have a single point of attachment to a core structure , such as an alkyl group , is identified in connection with a structure that must have two points of attachment in the structural core ( such as with the element l in formula ( ii )), it is understood that the named radical , e . g ., alkyl , refers to the parent radical with a hydrogen or a site of unsaturation removed to create the second point of attachment so as to provide the required structure . anionic groups according to the present invention , designated as “ a ” herein , include carboxylate , sulfate , sulfonate , sulfinate , sulfonamides , sulfones , phosphate , phosphonate , phosphinate , tetrazoles , thiophosphate , pyrophosphate , enolate , or a precursor group that can be converted to an anionic group , either spontaneously or enzymatically in cells . a specific embodiment of this invention is phenylglyoxal , shown below , although many other compounds that can achieve the desired effect are readily apparent to those skilled in the art , having regard for this disclosure : two additional categories of specific embodiments , in which an aliphatic chain is capable of binding to the active site of ftase by hydrophobic interactions , are defined as follows : also , substances with complex substituents might be better mimics of the farnesyl group of the substrate fpp and function more effectively as inhibitors , as for example , the compounds below : an additional compound that consists of the general formula r —( c ═ o )—( c ═ o )— r ′, or a masked form of such a group as denoted elsewhere in this application ( e . g . acetal , ketal , etc . ), and is contemplated by this disclosure is the following : the synthesis of the above compound may be carried out as follows : an additional compound that consists of the general formula r —( c ═ o )—( c ═ o )— r ′, or a masked form of such a group as denoted elsewhere in this application ( e . g . acetal , ketal , etc . ), and is contemplated by this disclosure is the following : the synthesis of the above compound may be carried out according to the following scheme : a further example of the class of compounds of the type r —( c ═ o )—( c ═ o )— r ′, or a masked form of such a group as denoted elsewhere in this application ( e . g . acetal , ketal , etc . ), is exemplified by dehydroascorbic acid 6 - palmitate , shown below : this compound may advantageously be synthesized by oxidation of ascorbic acid 6 - palmitate , as follows : other embodiments contemplated include compounds in which the —( ch 2 ) 14 ch 3 group shown in the structural formula of the 6 - palmitate is replaced by a group r as defined elsewhere herein . these include other 6 - esters of dehydroascorbic acid , such as dehydroascorbic acid 6 - farnesenoate , in which the —( ch 2 ) 14 ch 3 group shown in the structural formula of the 6 - palmitate is replaced by the — ch ═ c ( ch 3 ) ch 2 ch 2 ch ═ c ( ch 3 ) ch 2 ch 2 ch ═ c ( ch 3 ) 2 group . dehydroascorbic acid esters contemplated by this disclosure are inclusive of the 5 - isomers as well as the 6 - isomers . some cancer cells in which farnesylation of ras is blocked employ the related prenylation reaction geranylgeranylation to attach a hydrophobic group to ras to accomplish membrane localization and continued cancerous behavior of the cell . the enzyme that attaches the geranylgeranyl group to ras protein to facilitate the latter &# 39 ; s localization at the inner surface of the cancer cell membrane is geranylgeranyl protein transferase , also known as protein geranylgeranyltransferase ( herein referred to as ggtase ). the geranylgeranyl group becomes attached to ras by reaction with geranylgeranyl diphosphate , also known as geranylgeranyl pyrophosphate ( herein referred to as ggpp ). in other words , ggtase catalyzes the following reaction , in which ras becomes attached to the geranylgeranyl group by displacement of pyrophosphate ( pp i ): the newly formed geranylgeranyl - ras localizes at the inner surface of the cancer cell membrane and causes the cancer cell to divide without restraint . thus , a key target in a strategy to retard cancer cell proliferation is the enzyme ggtase . by reducing or destroying ggtase activity , either in combination with inactivation of ras farnesylation or independently , we hope to prevent ras geranylgeranylation , which in turn should prevent localization of ras at the inner surface of the cancer cell membrane , thereby preventing ras from causing the cancer cells to divide and proliferate . geranylgeranylation of ras results in the loss from ggpp of pyrophosphate , which is anionic , a process that might be facilitated by a chemical group in ggtase &# 39 ; s active site that would favorably interact with pyrophosphate , such as by ionic bonding to a positively charged group like the side chain of arginine [ r — nh ═ c ( nh 2 ) 2 + ]. thus , chemical reagents that react with the arginine side chain , such as those that possess the functional group consisting of two adjacent carbonyl groups , i . e ., o ═ cc ═ o , might effectively prevent one or more arginine residues in ggtase from assisting in the pyrophosphate departure , thereby blocking or impeding geranylgeranylation of ras . furthermore , to target better the substance to ggtase in preference to other enzymes , and to target better the substance to the active site of ggtase in preference to the external or noncatalytic portion of the enzyme , a preferred embodiment of the invention would consist of a substance with the dicarbonyl functional group and other structural features that would direct or otherwise favor the binding of the agent to the active site of ggtase . to fine - tune the inactivator to fit the active site of ftase or ggtase , variation of the distance between the arginine - binding end and the farnesyl - mimicking or geranylgeranyl - mimicking end can be achieved through alteration of the length of the spacer , i . e ., by variation of “ n ” from 0 to 10 in the generalized diagram below : additional inactivators incorporating aromatic groups for enhanced binding to the hydrophobic binding site of ftase or ggtase are exemplified by the following compound , although there are numerous possible variants of this type of compound ( e . g ., the location of the aromatic ring and the pattern of substitution as ortho , meta , or para ; a heterocyclic ring ; a nonaromatic ring ; multiple rings ; etc . ): a further embodiment of this invention is based on the observation that there are two arginine residues in the active site of ftase . because such arginine residues are at a characteristic distance unique to ftase , selective reaction with ftase over other enzymes and proteins can be accomplished by use of a substance in which two dicarbonyl functionalities are separated by a spacer , such substances being represented by the following generalized formula : in which d is a dicarbonyl functional group , o ═ c — c ═ o , or a masked form of such a group as denoted elsewhere in this application ( e . g . acetal , ketal , etc . ), and l is a linker group that is compatible with and / or has an affinity for the prenylation enzyme . one embodiment of this invention , in which two dicarbonyl functional groups are connected via a linker , is 4 , 4 ′- biphenyldiglyoxaldehyde , shown below : this category of compounds is further exemplified by the bis ( α - oxoaldehyde ) below : the spacer , ( ch 2 ) n , can be varied in length and in other characteristics ( e . g ., steric bulk , etc .) to optimize the selectivity for ftase and the ability to block the ftase active site . alternatives to the bis ( α - oxoaldehyde ) above include substances of the bis ( α - oxoketone ) type or compounds with both an α - oxoketone and α - oxoaldehyde functionality , as well as others readily apparent to those skilled in the art , having regard for this disclosure . thus , reaction of ftase with compounds such as the bis ( dicarbonyl ) compound shown above might occur with high efficiency and selectivity , and the covalent attachment of such compounds to ftase might be particularly effective at blocking the active site from utilizing the natural substrates fpp and ras protein , which need to bind to ftase in the vicinity of the two arginine residues . another embodiment of the invention utilizes the characteristics of the c - terminus of the ras protein , i . e ., the negative charge , for noncovalent binding to one of the arginine residues and the dicarbonyl functionality for covalent binding to the other arginine residue . the theory has been put forth that one of the arginine residues holds the ras protein in the active site through electrostatic binding of the arginine residue &# 39 ; s positive charge to the carboxyl terminus of ras protein , which is negatively charged . the other arginine residue binds fpp through the electrostatic attraction of the negative charge of fpp to the positive charge of the arginine side chain . thus , one such class of compounds that interfere with protein prenylation that is contemplated by this disclosure consists of four - part molecules symbolized as follows : a is ( 1 ) an anionic group , e . g ., carboxylate , sulfate , sulfonate , sulfinate , phosphate , phosphonate , thiophosphate , pyrophosphate , enolate , sulfonamide , sulfone , or tetrazole , ( 2 ) a group that can become anionic at physiological ph , e . g . co 2 h by deprotonation , or ( 3 ) a group that can be acted upon by an enzyme to reveal a group that can become anionic at physiological ph , e . g . the ester r ′ co 2 r is hydrolyzed by endogenous esterases to form the acid r ′ co 2 h , which subsequently ionizes at physiological ph to r ′ co 2 − ; l is a linker group that is compatible with and / or has an affinity for the prenylation enzyme , for example an alkyl , alkenyl , alkynyl , or aryl group . d is a dicarbonyl functional group ( o ═ c — c ═ o ) or a masked form of such a group as denoted elsewhere in this application ( e . g ., acetal , ketal , etc .). r is a group selected from the group consisting of an alkyl , alkenyl , alkynyl , or aryl group . specific embodiments of such a four - part molecule are the following camphorquinone sulfonamides : in which n is advantageously equal to 4 , 5 , 6 , or 7 , or another value for optimal binding to prenyltransferases . it will be apparent to those skilled in the art and having regard for this disclosure that other linking groups l , other anion - generating or anionic groups a , and other dicarbonyl groups d , or masked forms of such groups as denoted elsewhere in this application ( e . g . acetals , ketals , etc . ), can be advantageously substituted for the polymethylene chain —( ch 2 ) n —, carboxyl group , and camphorquinone moiety , respectively , in the above structure . for example , a prodrug , i . e ., a substance that has advantages of stability , solubility , cell permeability , etc ., but that liberates the active or a more active substance in the living tissues , e . g . spontaneously and / or by action of endogenous enzymes , light , and / or heat , could advantageously be formulated to be the camphorquinone sulfonamide esters represented by the structure shown below : in which r may be any of a very wide variety of groups , such as simple straight - chain , branched , or cyclic and heterocyclic aliphatic groups ( including functional groups containing heteroatoms , such as ether linkages , amino and hydroxyl groups ), and aromatic groups , including heterocycles . both of the enantiomeric camphorquinone sulfonamides ( i . e ., both (+) and (−) stereoisomers ) are contemplated by this patent and might be advantageously used for interference with protein prenylation . the synthesis of camphorquinone sulfonamides can be carried out as illustrated below : additionally , the compound below , when in its ionized form ( the predominant form present at physiological ph ), has the characteristics required for the noncovalent binding of the carboxylate terminus to one arginine residue , and the covalent binding of the dicarbonyl functionality to the other arginine residue : when m = 9 , the compound is 11 , 12 - dioxododecanoic acid , a preferred compound of the invention . typically m = 1 to 25 , more preferably 5 - 13 . other embodiments can combine the carboxylic acid group and an α - oxoaldehyde or , alternatively , a carboxylic acid group and an α - oxoketone group within the same molecule , or other combinations that are readily apparent to those skilled in the art , having regard for this disclosure . by modification of the linker group &# 39 ; s structure ( e . g ., length , steric bulk , etc . ), the specificity of the compound for ftase and the effectiveness of the compound against ftase can be fine - tuned . use of known non - covalent inhibitors of ftase and ggtase as linker or r groups is preferred to increase specificity for the enzyme active site . for example , the tetrapeptide portion of ras that is recognized by ftase ( referred to as caax , in which c represents cysteine , a represents an aliphatic amino acid , and x represents a limited subset of the naturally occurring amino acids , of which serine is an example ) can be utilized in combination with a dicarbonyl functionality to target the inactivator to its target , ftase . such a compound might be a useful inactivator . it might , however , be hydrolyzed by cellular proteases , so a peptidomimetic analogue less prone to cellular hydrolysis might show superior lifetime in a cell , thereby enhancing its chances of combining with ftase and inactivating it . an embodiment of a peptidomimetic analogue of the tetrapeptide in which a dicarbonyl functionality is present is shown below : in the structure above , r 1 is a substituent suitable for recognition and binding to ftase , and r 2 is typically h or ch 3 , for reaction with an arginine residue in the ftase active site . many other embodiments of this type are readily apparent to those skilled in the art , having regard for this disclosure . there are numerous ways to synthesize a - dicarbonyl compounds . one is by oxidation of a monocarbonyl compound with selenium dioxide ( seo 2 ), shown below : another method of preparation of α - dicarbonyl compounds is by alkylation of 2 , 3 - butanedione , illustrated below : another means of synthesis of α - dicarbonyl compounds is illustrated below , starting with citronellal : other embodiments of the invention are substances in which one or more carbonyl groups are masked as a hydrate [ c ( oh ) 2 ], a hemiacetal or hemiketal [ c ( oh )( or ″)], an acetal or a ketal [ c ( or ″)( or ′″)], an acylal or related compound [ c ( oc (═ o ) r ″)( oc (═ o ) r ′″)], a bisulfite addition compound [ c ( oh )( so 3 — )], an enol ( c ═ coh ), an enol ether ( c ═ cor ″), an enol ester [ c ═ coc (═ o ) r ″], and so forth , wherein r ″ and r ′″, which may be the same or different , are alkyl , alkenyl , and / or aryl groups . such masked carbonyl groups may produce the carbonyl form of the reagent in solution . acetals and ketals with hydrophobic side chains , including those that resemble the farnesyl group , are shown below : the examples below show that ftase is sensitive to inactivation by phenylglyoxal and by 2 - oxododecanal , both of which are α - dicarbonyl compounds . new opportunities exist for the development of - dicarbonyl compounds and related substances that might prove to be inactivators of prenylation enzymes with varying potencies and specificities . a specific protein designated k - ras is the most common mutant ras protein found in human cancers . farnesylation inhibitors , however , cannot prevent cellular prenylation of k - ras because geranylgeranylation of k - ras takes place . if ggtase has an active - site arginine , - dicarbonyl compounds like pgo might likewise be effective inactivators of that enzyme as well . the ability of the α - dicarbonyl functional group to achieve ftase inactivation is described in the examples below , and compounds of this type offer a new approach to the treatment and control of cancer . suitable levels of enzyme activity reduction range from at least 10 % to at least 95 % or even 100 %. any level can be selected as desired , using appropriate chemical substitutents to achieve at least 10 , 20 , 30 , 40 , 50 , 60 , 70 , 80 , or 90 % reduction in enzyme activity . typical routes of administration of these substances that might be suitable for clinical applications are oral and intravenous , although other routes might prove beneficial in specific cancers ( e . g ., intraperitoneal , intramuscular , subcutaneous , intrathecal , or topical for skin cancers ). doses and frequency of administration are to be selected based upon considerations of beneficial effects ( e . g ., tumor growth reduction or tumor shrinkage ) compared to side effects ( e . g ., systemic toxicity ). combination with other anticancer agents to produce synergistic effects of benefit to the patient are also possible . combination therapy might be based on two strategies . one is to interfere with different biochemical processes to increase tumor cell killing . another is to hamper development of drug resistance , which is less likely to occur simultaneously in tumor cells exposed to anticancer agents based on interference with different biochemical pathways in the tumor cells . compounds according to the invention may be purified free of impurities , preferably to a level of at least 50 , 70 , 90 , or 95 %. the compounds may be formulated in suitable pharmaceutically acceptable diluents , excipients or carriers . these can provide for additional properties , such as enhanced absorption , slow release , tissue or organ targeting , etc . in summary an improved method of interference with protein prenylation in tumor cells has been described that may prevent or hamper the proliferation of tumor cells , possibly resulting in a decrease in tumor size and / or disappearance of the cancer . an example of an embodiment of this invention is phenylglyoxal monohydrate , shown below , which is in equilibrium with the dicarbonyl form : the application of phenylglyoxal monohydrate to solutions of ftase resulted in the rapid and virtually complete inactivation of the enzyme in a matter of seconds to minutes , depending upon the concentration of phenylglyoxal monohydrate applied . for example , the catalytic ability of ftase was monitored by an assay procedure in which ftase transferred the farnesyl group from fpp to a peptide substitute for ras . the peptide substitute for ras in this assay had structural elements in common with ras , in particular a sequence of four amino acids cysteinyl - valyl - lysyl - serine ( cys - val - lys - ser ). in addition the peptide contained an amino acid residue that had a fluorescent marker attached , dansyl - glycine ( dns - gly ). farnesyl group transfer by ftase from fpp to dns - gly - cys - val - lys - ser resulted in an increase in fluorescence due to the attachment of the hydrophobic group to the cysteine near the dansyl group . thus , enzyme activity is revealed by an increase in fluorescence of the dansyl group , as shown in the accompanying fig1 . inactivation of the enzyme ftase occurred when the ftase was preincubated with phenylglyoxal monohydrate , as evidenced by ( 1 ) a decrease in enzyme activity with increasing time of preincubation with phenylglyoxal monohydrate ; and ( 2 ) a decrease in enzyme activity with increasing concentration of phenylglyoxal monohydrate . because of interference of phenylglyoxal monohydrate with the fluorescence assay used , phenylglyoxal monohydrate was removed from the ftase preincubation mixture by ultrafiltration prior to the assay being performed . the accompanying fig2 shows the loss of activity of ftase due to inactivation by phenylglyoxal monohydrate . evidence for the binding of phenylglyoxal in the farnesyl pyrophosphate binding region of the enzyme was obtained by preincubation of the enzyme with farnesyl pyrophosphate , followed by treatment with phenylglyoxal . as shown in the accompanying fig3 , higher concentrations of farnesyl pyrophosphate more effectively reduced the inactivation of ftase by phenylglyoxal . this suggests that the farnesyl pyrophosphate bound to and blocked phenylglyoxal from the site of the reaction that brings about inactivation , namely chemical modification of an active - site arginine side chain . a detailed description of the results summarized above follows . upon farnesylation by protein farnesyltransferase ( ftase ), 1 ras protein becomes localized at the inner surface of the cell membrane . localization is essential for the mitogenic role of mutant ras protein , which acts as a switch for cancer cell proliferation . we found that phenylglyoxal ( pgo ), which is a protein - modification reagent that is specific for arginine , potently obstructs farnesylation of a ras model peptide by ftase . covalent modification interferes with catalysis and results in mechanism - based inactivation of ftase . ftase activity in vitro was monitored by a fluorescence assay . ftase was rapidly inactivated by preincubation with pgo . at 20 mm pgo , ftase activity was totally lost within 2 minutes . inactivation rates at 10 mm pgo and 5 mm pgo were successively slower . preincubation of ftase with fpp prior to incubation with pgo decreased the ability of pgo to inactivate ftase . these results imply that a bimolecular reaction occurred between pgo and ftase , probably in the vicinity of the fpp binding site , which resulted in the inactivation of the enzyme . these findings reveal the value of - dicarbonyl compounds for mechanistic studies of ftase and for potentially selective regulation of protein farnesylation for possible anticancer chemotherapy . footnotes : 1 the abbreviations used are : ds - gcvls , the dansyl - labeled pentapeptide n - dimethylaminonaphthalenesulfonyl - gly - cys - val - leu - ser ; dtt , dithiothreitol ; fmoc , 9 - fluorenylmethoxycarbonyl ; ftase , human recombinant protein farnesyltransferase ; fpp , farnesyl pyrophosphate ; ggpp , geranylgeranyl pyrophosphate ; ggtase , protein geranylgeranyltransferase ; hepes , n - 2 - hydroxyethylpiperazine - n - 2 - ethanesulfonic acid ; pgo , phenylglyoxal monohydrate ; tris , tris ( hydroxymethyl ) aminomethane . posttranslational modification of a small number of proteins is carried out by protein farnesyltransferase ( ftase ), which attaches the 15 - carbon farnesyl group of farnesyl pyrophosphate to the sulfhydryl group of a cysteine residue near the c - terminus of the substrate protein . noteworthy among the proteins that are substrates for ftase is ras protein , which upon farnesylation becomes localized at the lipid bilayer of the inner surface of the cell membrane . localization is essential for the mitogenic role of ras , which acts as a switch for cell division . cycling between the inactive form ( gdp - bound ras ) and the active form ( gtp - bound ras ), is part of the normal mechanism of cell growth regulation . mutant forms of ras protein have lost gtpase activity and trigger unrestrained cell division . approximately one third of all human cancers exhibit a mutant ras genotype , with substantial variations in the incidence from tumor type to tumor type . this has made blockage of farnesylation of mutant ras protein an active area of research as it appears that inhibition of both normal and mutant ras protein farnesylation is more harmful to cancer cells than normal cells . the challenges to this coup de main against cancer , however , are manifold . some cancer cell lines with mutant ras genotypes are insensitive to ftase inhibitors , and some cancer cell lines without mutant ras genotypes are sensitive to ftase inhibitors . this may mean that a delicate balance between reduction in farnesylation of different proteins is required for effective anticancer strategies . a possible addition to the armamentarium of farnesylation regulators based on a new approach is described in this report . recognition of ras and related protein substrates of ftase is based on a structural motif consisting of a c - terminal caax sequence , in which a represents an aliphatic amino acid and x is preferentially serine . the related posttranslational geranylgeranylation , in which the 20 - carbon geranylgeranyl chain is attached to the cysteine of caax by geranylgeranyltransferase ( ggtase ), occurs preferentially when x is methionine . nucleophilic attack by the cysteine sulfhydryl group of caax , activated by an enzyme - bound zinc ion , results in ras farnesylation , with departure of pyrophosphate from fpp . we surmised that catalysis of farnesylation by ftase might be facilitated by salt - bridge formation ( i . e ., electrostatic stabilization ) between the pyrophosphate leaving group of farnesyl pyrophosphate and a cationic amino acid side chain such as the guanidinium group of arginine . this speculation was strengthened by the recent publication of some aspects of ftase structure obtained from x - ray crystallographic analyses . an arginine ( arg 291 ) was found to be in close proximity to the binding site of the diphosphate portion of fpp and the active - site zinc ion , near the upper part of the barrel - shaped active site . another key arginine in the bottom of the active site , arg 202 , is proposed to form a salt bridge with the carboxyl group of the caax terminus of the ras peptide substrate . in addition the aliphatic part of the arginine side chain is buttressed with the fpp aliphatic chain . ftase &# 39 ; s preference for fpp relative to ggpp can be understood because the latter would project from the cleft and be poorly positioned for transfer of the geranylgeranyl group to the protein substrates of ftase . we report herein that a protein - modification agent , phenylglyoxal , that is specific for the arginine side chain potently obstructs farnesylation of a ras model peptide by ftase . this approach of specific covalent modification of ftase to interfere with catalysis results in mechanism - based inactivation of ftase , in contrast to the numerous noncovalent inhibitors that have been identified . covalent modification is in principle virtually irreversible and might be tailored to active site residues of ftase . inactivation thus opens the door to mechanistic studies and potentially selective obstruction of ftase and / or ggtase for possible anticancer chemotherapeutic benefit . materials . all reagents were from sigma chemical co . and used without further purification , except as noted . purification of pgo — pgo ( aldrich chemical co .) was dissolved in boiling water that contained charcoal , and the mixture was gravity filtered through celite while hot . upon cooling , the solution was filtered by suction to obtain the pgo , and the pgo was repeatedly recrystallized from water until its 1 h nmr spectrum and melting point indicated the absence of impurities , such as polymeric material . all pgo solutions were freshly prepared before use in the studies described below . preparation of ds - gcvls ds - gcvls was prepared by a method analogous to a published procedure . synthesis was performed by use of a millipore 9050 plus pepsynthesizer that employed standard fmoc solid phase peptide synthesis methodology . ps - peg resin and fmoc protected amino acids were obtained from millipore . the n - dansylglycine was from sigma chemical co . peptide purification was performed by rp - hplc on an alltech macrosphere rp300 c8 column . elution was accomplished with a linear gradient from aqueous 10 mm trifluoroacetic acid to 10 mm trifluoroacetic acid in acetonitrile . lyophilization yielded a pale yellow solid . stock solutions were prepared in 50 mm tris containing 5 mm dtt and 0 . 2 % n - octyl - β - d - glucopyranoside ( anatrace , inc . ), ph 7 . 7 . concentrations of ds - gcvls were determined by use of the extinction coefficient of the dansyl group at 340 nm (= 4250 m − 1 cm − 1 ). stock solutions were stored at − 20 c . ftase ( stored at − 70 c .) was a generous gift from professor patrick casey at duke university . ftase assay the fluoresence assay conditions were adapted from a published method . the ftase assay consisted of the measurement of the rate of ftase - catalyzed farnesylation of the fluorescent pentapeptide ds - gcvls . upon farnesylation of the cysteine , the fluoresence emission at = 505 nm of the dansyl group is enhanced due to the proximity of the hydrophobic isoprenoid group . enzyme activity ( approximately 60 nm ftase ) was monitored at saturating substrate concentrations ( 10 m farnesyl pyrophosphate , ammonium salt , prepared by successive dilutions of a commercial 2 . 3 mm solution in methanol : 10 mm aqueous nh 4 oh ( 7 : 3 ) into assay buffer , and 1 . 0 m ds - gcvls ) by measurement of the increase in fluorescence emission observed at = 505 nm as ftase farnesylated ds - gcvls for a period of 10 min . the assay buffer was selected for unreactivity toward pgo : 50 mm nahepes , 5 mm dtt , 5 mm mgcl 2 , 10 μm zncl 2 , and 0 . 2 % n - octyl - β - d - glucopyranoside , ph 7 . 5 . fluorescence emission was measured with a jasco fp - 77 spectrofluorimeter fitted with a temperature - controlled cuvette holder connected to a 30 c . constant temperature bath . all fluoresence assays were conducted in a 4 mm 4 mm quartz cuvette with excitation at 340 nm . inactivation of ftase by pgo ftase was incubated at 30 c . in the presence and absence of pgo at various concentrations in 50 mm nahepes , 5 mm mgcl 2 , 10 μm zncl 2 , and 0 . 2 % n - octyl - β - d - glucopyranoside , ph 7 . 5 ( i . e ., assay buffer without dtt ). periodically , aliquots of the ftase - pgo reaction mixture were removed and assayed for ftase activity . because of interference of pgo with the fluorometric assay , the aliquot was first subjected to solvent exchange by size exclusion membrane concentration to remove pgo . the solvent exchange utilized microcon ( millipore co .) 30 kd molecular weight cutoff microconcentrators . this also served to stop further inactivation of the enzyme by pgo . a 100 - l aliquot of ftase - pgo reaction mixture was pipetted into a microcon concentrator , which was centrifuged at 12 , 000 rpm for 8 min at 4 c . the concentrate was resuspended in 100 l of assay buffer , and centrifugation was repeated . the final concentrate was resuspended in 90 l of assay buffer , and the resulting solution was recovered by inversion of the concentrator and centrifugation at 3500 rpm for 3 min at 4 c . the solution was then assayed for ftase activity as described above . a control was handled as above , except that pgo was omitted from the incubation mixture . fpp protection of ftase against inactivation by pgo ftase was treated with fpp at various concentrations in assay buffer without dtt for 2 min at 30 ° c . prior to incubation of the mixture with 10 mm pgo for 30 min at 30 ° c . the control was incubated with fpp at the corresponding concentration for the full length of time , but without pgo . the mixtures were subsequently subjected to microconcentration and assay , as described above . ftase activity in vitro was monitored by a fluorescence assay , shown in fig1 , in which activity was determined from the initial slope of a plot of the fluorescence increase vs . time . ftase was rapidly inactivated by preincubation with pgo , as shown in fig2 . at 20 mm pgo , ftase activity was totally lost before the first assay at 2 minutes after mixing of pgo and ftase ( data not shown ). inactivation by 10 mm pgo was slower but still quite rapid . at 5 mm pgo the rate of inactivation was reduced . these results imply that a bimolecular reaction occurred between pgo and ftase , which resulted in the inactivation of the enzyme . inactivation kinetics differ from inhibition kinetics . inactivation kinetics typically consist of the bimolecular conversion of the active form of the enzyme ( e ) into impaired and / or completely inactive form ( s ) of the enzyme ( e ′), as shown in the equation below , where pgo is the inactivator : because the inactivator is typically present in large excess over the enzyme and therefore remains at essentially constant concentration , disappearance of enzyme activity follows pseudo - first - order kinetics , described by the following equation : in which [ e ] t is the enzyme concentration or activity remaining at time t , [ e ] 0 is the initial enzyme concentration or activity , and [ pgo ] is the concentration of pgo . thus , a plot of log [ e ] t /[ e ] 0 , i . e ., log ( fraction of activity remaining ), versus time is expected to be linear , with a slope of k [ pgo ]/ 2 . 303 . thus , the time - dependence and inhibitor - concentration - dependence of loss of enzyme activity are indicative of inactivation of the enzyme . previously , the stepwise formation of a 2 : 1 pgo - enzyme adduct has been seen kinetically as a biphasic time course of inactivation of porcine liver prenyltransferase , presumably due to rapid , reversible 1 : 1 adduct formation resulting in reduced activity of the enzyme , followed by slower , irreversible 2 : 1 adduct formation , and conversion of the enzyme into a less active form . some indication of similar behavior is seen in fig2 , especially at low pgo concentration . preincubation of ftase with fpp prior to incubation with pgo decreased the ability of pgo to inactivate ftase , as shown in fig3 . this implies that an active site arginine residue is the principal site of the reaction with pgo that inactivates the enzyme . when arginine modification occurs in the fpp binding site , then obstruction of subsequent binding of fpp would be expected to prevent farnesylation of the substrate pentapeptide . the low concentration of pgo required for inactivation of the enzyme is also consistent with the idea that specific modification of an active - site arginine has occurred . it is not possible to rule out that the arginine side - chain modification results in destruction of the catalytic ability of the active - site arginine without blockage of fpp binding . likewise , some modification of lysine side chains and - amino groups is possible with phenylglyoxal , although under the short incubation times used in these studies , such modification would be expected to be of minor importance . destruction of enzyme activity by modification of numerous arginines , which might for example lead to gross conformational changes and concomitant loss of catalytic ability , also would not be expected to occur under these conditions . in either case , the observed protective effect of fpp would not necessarily have been expected . future studies , however , are required to establish whether arg 291 is the principal site of modification of ftase by pgo . such studies will also ascertain whether another amino acid is a significant target of pgo modification . in the same manner as example 1 , compound 2 - oxododecanal was tested for inactivation of ftase in the fluorescence assay system described , except as noted below . final concentrations in the incubation mixture were 5 , 10 , and 35 mm 2 - oxododecanal . a stock solution of 2 - oxododecanal dissolved in dimethyl sulfoxide ( dmso ) was employed . the final concentration of dmso in the incubation mixture ( as well as the control ) was 5 %. the results are shown in fig4 . it was found that 2 - oxododecanal rapidly inactivated ftase , probably by reaction with one or both of the active site arginine residues . this would block access of one or both of the substrates ( fpp and / or the peptide substitute for ras protein ) and / or block the catalytic function of one or both of the arginine residues , resulting in inactivation of the enzyme . in the same manner as example 1 , compound 5 , 9 - dimethyl - 8 - decene - 2 , 3 - dione was tested for inactivation of ftase in the fluorescence assay system described , except as noted below . a stock solution of 5 , 9 - dimethyl - 8 - decene - 2 , 3 - dione in 5 % dmso was employed . the final concentration of dmso in the incubation mixture ( as well as the control ) was 4 . 6 %. ftase was at 15 nm , and the microcon concentration step was omitted . it was found that 5 , 9 - dimethyl - 8 - decene - 2 , 3 - dione rapidly inactivated ftase . after 30 minutes of incubation with 17 , 68 , or 171 μm 5 , 9 - dimethyl - 8 - decene - 2 , 3 - dione , ftase activity was reduced to 38 %, 18 or 16 % of the initial control value , respectively . in the same manner as example 1 , dehydroascorbic acid 6 - palmitate was tested for inactivation of ftase in the fluorescence assay system described , except as noted below . a stock solution of dehydroascorbic acid 6 - palmitate in 2 % dmso in 5 mm sodium acetate , ph 3 . 9 , was employed . the final concentration of dmso in the incubation mixture ( as well as the control ) was 0 . 46 %. ftase was at 15 nm , and the microcon concentration step was omitted . it was found that dehydroascorbic acid 6 - palmitate rapidly inactivated ftase . after 30 minutes of incubation with 7 , 28 , or 141 μm dehydroascorbic acid 6 - palmitate , ftase activity was reduced to 75 %, 34 %, or 22 % of the initial control value , respectively . in the same manner as example 1 , compound 4 , 4 ′- biphenyldiglyoxaldehyde was tested for inactivation of ftase in the fluorescence assay system described , except as noted below . a stock solution of 4 , 4 ′- biphenyldiglyoxaldehyde in 5 % dmso was employed . the final concentration of dmso in the incubation mixture ( as well as the control ) was 4 . 6 %, and ftase was at 15 nm . it was found that 4 , 4 ′- biphenyldiglyoxaldehyde rapidly inactivated ftase . after 30 minutes of incubation with 23 or 93 μm 4 , 4 ′- biphenyldiglyoxaldehyde , ftase activity was reduced to 68 % or 11 % of the initial control value , respectively . as a test of the importance of the presence of a dicarbonyl functional group , a compound analogous to that used in example 5 but without such a group was tested . thus , in the same manner as example 1 , compound 4 , 4 ′- diacetylbiphenyl , whose structure is as follows : was tested for inactivation of ftase in the fluorescence assay system described , except as noted below . a stock solution of 4 , 4 ′- diacetylbiphenyl in 5 % dmso was employed . the final concentration of dmso in the incubation mixture ( as well as the control ) was 4 . 6 %, ftase was at 15 nm . it was found that 4 , 4 ′- diacetylbiphenyl did not inactivated ftase . even after 120 minutes of incubation with 23 μm 4 , 4 ′- diacetylbiphenyl , ftase activity was the same as the control , within experimental error . in the same manner as example 1 , compound 2 , 3 - pentanedione , ch 3 ch 2 ( c ═ o )—( c ═ o ) ch 3 , was tested for inactivation of ftase in the fluorescence assay system described , except as noted below . the assay was carried out in the presence of 50 mm sodium borate , ph 7 . 5 . ftase was at 15 nm , and the microcon concentration step was omitted . it was found that 2 , 3 - pentanedione rapidly inactivated ftase . after approximately 30 minutes of incubation with 17 , 33 , or 66 mm 2 , 3 - pentanedione , ftase activity was reduced to 69 %, 55 %, or 37 % of the initial control value , respectively . 3 . casey , p . j . and seabra , m . c . ( 1996 ) j . biol . chem . 271 , 5289 - 5292 . 4 . leonard , d . m . ( 1997 ) j . med . chem . 40 , 2971 - 2990 . 5 . gibbs , j . b . and oliff , a . ( 1997 ) annu . rev . pharmacol . toxicol . 37 , 143 - 166 . 6 . cassidy , p . b . and poulter , c . d . ( 1996 ) j . am . chem . soc . 118 , 8761 - 8762 . 7 . patel , d . v ., young , m . g ., robinson , s . p ., hunihan , l ., dean , b . j ., and gordon , e . m . ( 1996 ) j . med . chem . 39 , 4197 - 4210 . 8 . sattler , i . and tamanoi , f . ( 1996 ) “ prenylation of ras and inhibitors of prenyltransferases .” in regulation of the ras signaling network , h . maruta and a . w . burgess , eds ., chapter 4 , chapman & amp ; halkl , austin , tex . 9 . huang , c .- c ., casey , p . j ., and fierke , c . a . ( 1997 ) j . biol . chem . 272 , 20 - 23 . 10 . dolence , j . m ., cassidy , p . b ., mathis , j . r ., and poulter , c . d . ( 1996 ) biochemistry 34 16687 - 16694 . 11 . mu , y . q ., omer , c . a ., and gibbs , r . a . ( 1996 ) j . am . chem . soc . 118 , 1817 - 1823 . 12 . park , h .- w ., boduluri , s . r ., moomaw , j . f ., casey , p . j ., and beeese , l . s . ( 1997 ) science 275 , 1800 - 1804 ; dunten , p ., kammlott , u ., crowther , r ., weber , d ., palermo , r ., and birktoft , j . ( 1998 ) biochemistry 37 , 7907 - 7912 ; long , s . b ., casey , p . j , and beese , l . s . ( 1998 ) biochemistry 37 , 9612 - 9618 . 13 . takahashi , k . ( 1968 ) j . biol . chem . 243 , 6171 - 6179 . 14 . boivin , d ., lin , w ., and beliveau , r . ( 1997 ) biochem . cell biol . 75 , 63 - 69 . 15 . barnard , g . f . and popjak , g . ( 1980 ) biochim . biophys . acta 617 , 169 - 182 . 16 . pompliano , d . l ., gomez , r . p ., and anthony , n . j . ( 1992 ) j . am . chem . soc . 114 , 7945 - 7946 . 17 . patthy , l . and thesz , j . ( 1980 ) eur . j . biochem . 105 , 387 - 393 . 18 . rowell , c . a ., kowalczyk , j . j ., lewis , m . d ., and garcia , a . m . ( 1997 ) j . biol . chem . 272 , 14093 - 14097 . 19 . whyte , d . b ., kirschmeier , p ., hockenberry , t . n ., nunez - oliva , i ., james , l ., catino , j . j ., bishop , w . r ., and pai , j .- k . ( 1997 ) j . biol . chem . 272 , 14459 - 14464 .

prenylating enzymes are involved in modifying oncoproteins , such as ras , so that growth of neoplastic cells becomes uncontrolled . inactivation of such enzymes can prevent uncontrolled growth . α - dicarbonyl compounds can be used to covalently modify and thereby inactivate prenylating enzymes such as protein farnesyltransferase and protein geranylgeranyltransferase . the compounds can be designed to enhance affinity and / or specificity for a particular protein substrate .

“ aerodynamic diameter ” of a given particle refers to the diameter of a spherical droplet with a density of 1 g / ml ( the density of water ) that has the same settling velocity as the given particle . “ aerosol ” refers to a suspension of solid or liquid particles in a gas . “ aerosol drug mass density ” refers to the mass of antidepressant per unit volume of aerosol . “ aerosol mass density ” refers to the mass of particulate matter per unit volume of aerosol . “ aerosol particle density ” refers to the number of particles per unit volume of aerosol . “ amorphous particle ” refers to a particle that does not contain more than 50 percent by weight of a crystalline form . preferably , the particle does not contain more than 25 percent by weight of a crystalline form . more preferably , the particle does not contain more than 10 percent by weight of a crystalline form . “ amoxapine degradation product ” refers to a compound resulting from a chemical modification of amoxapine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ antidepressant degradation product ” refers to a compound resulting from a chemical modification of an antidepressant . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ bupropion degradation product ” refers to a compound resulting from a chemical modification of bupropion . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ citalopram degradation product ” refers to a compound resulting from a chemical modification of citalopram . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ clomipramine degradation product ” refers to a compound resulting from a chemical modification of clomipramine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ condensation aerosol ” refers to an aerosol formed by vaporization of a substance followed by condensation of the substance into an aerosol . “ doxepin degradation product ” refers to a compound resulting from a chemical modification of doxepin . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ fluoxetine degradation product ” refers to a compound resulting from a chemical modification of fluoxetine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . examples of fluoxetine degradation products include 4 -( trifluoromethyl )- phenol and 3 - phenyl - 2 - propenal . “ fluvoxamine degradation product ” refers to a compound resulting from a chemical modification of fluvoxamine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ imipramine degradation product ” refers to a compound resulting from a chemical modification of imipramine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ inhalable aerosol drug mass density ” refers to the aerosol drug mass density produced by an inhalation device and delivered into a typical patient tidal volume . “ inhalable aerosol mass density ” refers to the aerosol mass density produced by an inhalation device and delivered into a typical patient tidal volume . “ inhalable aerosol particle density ” refers to the aerosol particle density of particles of size between 100 nm and 5 microns produced by an inhalation device and delivered into a typical patient tidal volume . “ mass median aerodynamic diameter ” or “ mmad ” of an aerosol refers to the aerodynamic diameter for which half the particulate mass of the aerosol is contributed by particles with an aerodynamic diameter larger than the mmad and half by particles with an aerodynamic diameter smaller than the mmad . “ maprotiline degradation product ” refers to a compound resulting from a chemical modification of maprotiline . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ mirtazepine degradation product ” refers to a compound resulting from a chemical modification of mirtazepine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ nefazodone degradation product ” refers to a compound resulting from a chemical modification of nefazodone . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ nortriptylene degradation product ” refers to a compound resulting from a chemical modification of nortriptylene . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ paroxetine degradation product ” refers to a compound resulting from a chemical modification of paroxetine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ perphenazine degradation product ” refers to a compound resulting from a chemical modification of perphenazine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ protriptylene degradation product ” refers to a compound resulting from a chemical modification of protriptylene . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ rate of aerosol formation ” refers to the mass of aerosolized particulate matter produced by an inhalation device per unit time . “ rate of inhalable aerosol particle formation ” refers to the number of particles of size between 100 nm and 5 microns produced by an inhalation device per unit time . “ rate of drug aerosol formation ” refers to the mass of antidepressant produced by an inhalation device per unit time . “ sertraline degradation product ” refers to a compound resulting from a chemical modification of sertraline . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ settling velocity ” refers to the terminal velocity of an aerosol particle undergoing gravitational settling in air . “ trazodone degradation product ” refers to a compound resulting from a chemical modification of trazodone . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ tranylcypromine degradation product ” refers to a compound resulting from a chemical modification of tranylcypromine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ trimipramine degradation product ” refers to a compound resulting from a chemical modification of trimipramine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ typical patient tidal volume ” refers to 1 l for an adult patient and 15 ml / kg for a pediatric patient . “ vapor ” refers to a gas , and “ vapor phase ” refers to a gas phase . the term “ thermal vapor ” refers to a vapor phase , aerosol , or mixture of aerosol - vapor phases , formed preferably by heating . “ valproic acid degradation product ” refers to a compound resulting from a chemical modification of valproic acid . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . “ venlafaxine degradation product ” refers to a compound resulting from a chemical modification of venlafaxine . the modification , for example , can be the result of a thermally or photochemically induced reaction . such reactions include , without limitation , oxidation and hydrolysis . any suitable method is used to form the aerosols of the present invention . a preferred method , however , involves heating a composition comprising an antidepressant to form a vapor , followed by cooling of the vapor such that it condenses to provide an antidepressant comprising aerosol ( condensation aerosol ). the composition is heated in one of four forms : as pure active compound ( e . g ., pure bupropion , nefazodone , perphenazine , trazodone , trimipramine , venlafaxine , tranylcypromine , citalopram , fluoxetine , fluvoxamine , mirtazepine , paroxetine , sertraline , amoxapine , clomipramine , doxepin , imipramine , maprotiline , nortryptiline , valproic acid , or protryptyline ); as a mixture of active compound and a pharmaceutically acceptable excipient ; as a salt form of the pure active compound ; and , as a mixture of active compound salt form and a pharmaceutically acceptable excipient . salt forms of antidepressants ( e . g ., bupropion , nefazodone , perphenazine , trazodone , trimipramine , venlafaxine , tranylcypromine , citalopram , fluoxetine , fluvoxamine , mirtazepine , paroxetine , sertraline , amoxapine , clomipramine , doxepin , imipramine , maprotiline , nortryptiline , valproic acid , or protryptyline ) are either commercially available or are obtained from the corresponding free base using well known methods in the art . a variety of pharmaceutically acceptable salts are suitable for aerosolization . such salts include , without limitation , the following : hydrochloric acid , hydrobromic acid , acetic acid , maleic acid , formic acid , and fumaric acid salts . pharmaceutically acceptable excipients may be volatile or nonvolatile . volatile excipients , when heated , are concurrently volatilized , aerosolized and inhaled with the antidepressants . classes of such excipients are known in the art and include , without limitation , gaseous , supercritical fluid , liquid and solid solvents . the following is a list of exemplary carriers within the classes : water ; terpenes , such as menthol ; alcohols , such as ethanol , propylene glycol , glycerol and other similar alcohols ; dimethylformamide ; dimethylacetamide ; wax ; supercritical carbon dioxide ; dry ice ; and mixtures thereof . solid supports on which the composition is heated are of a variety of shapes . examples of such shapes include , without limitation , cylinders of less than 1 . 0 mm in diameter , boxes of less than 1 . 0 mm thickness and virtually any shape permeated by small ( e . g ., less than 1 . 0 mm - sized ) pores . preferably , solid supports provide a large surface to volume ratio ( e . g ., greater than 100 per meter ) and a large surface to mass ratio ( e . g ., greater than 1 cm 2 per gram ). a solid support of one shape can also be transformed into another shape with different properties . for example , a flat sheet of 0 . 25 mm thickness has a surface to volume ratio of approximately 8 , 000 per meter . rolling the sheet into a hollow cylinder of 1 cm diameter produces a support that retains the high surface to mass ratio of the original sheet but has a lower surface to volume ratio ( about 400 per meter ). a number of different materials are used to construct the solid supports . classes of such materials include , without limitation , metals , inorganic materials , carbonaceous materials and polymers . the following are examples of the material classes : aluminum , silver , gold , stainless steel , copper and tungsten ; silica , glass , silicon and alumina ; graphite , porous carbons , carbon yams and carbon felts ; polytetrafluoroethylene and polyethylene glycol . combinations of materials and coated variants of materials are used as well . where aluminum is used as a solid support , aluminum foil is a suitable material . examples of silica , alumina and silicon based materials include amphorous silica s - 5631 ( sigma , st . louis , mo . ), bcr171 ( an alumina of defined surface area greater than 2 m 2 / g from aldrich , st . louis , mo .) and a silicon wafer as used in the semiconductor industry . carbon yams and felts are available from american kynol , inc ., new york , n . y . chromatography resins such as octadecycl silane chemically bonded to porous silica are exemplary coated variants of silica . the heating of the antidepressant compositions is performed using any suitable method . examples of methods by which heat can be generated include the following : passage of current through an electrical resistance element ; absorption of electromagnetic radiation , such as microwave or laser light ; and , exothermic chemical reactions , such as exothermic solvation , hydration of pyrophoric materials and oxidation of combustible materials . antidepressant containing aerosols of the present invention are delivered to a mammal using an inhalation device . where the aerosol is a condensation aerosol , the device has at least three elements : an element for heating an antidepressant containing composition to form a vapor ; an element allowing the vapor to cool , thereby providing a condensation aerosol ; and , an element permitting the mammal to inhale the aerosol . various suitable heating methods are described above . the element that allows cooling is , in it simplest form , an inert passageway linking the heating means to the inhalation means . the element permitting inhalation is an aerosol exit portal that forms a connection between the cooling element and the mammal &# 39 ; s respiratory system . one device used to deliver the antidepressant containing aerosol is described in reference to fig1 . delivery device 100 has a proximal end 102 and a distal end 104 , a heating module 106 , a power source 108 , and a mouthpiece 110 . an antidepressant composition is deposited on a surface 112 of heating module 106 . upon activation of a user activated switch 114 , power source 108 initiates heating of heating module 106 ( e . g , through ignition of combustible fuel or passage of current through a resistive heating element ). the antidepressant composition volatilizes due to the heating of heating module 106 and condenses to form a condensation aerosol prior to reaching the mouthpiece 110 at the proximal end of the device 102 . air flow traveling from the device distal end 104 to the mouthpiece 110 carries the condensation aerosol to the mouthpiece 110 , where it is inhaled by the mammal . devices , if desired , contain a variety of components to facilitate the delivery of antidepressant containing aerosols . for instance , the device may include any component known in the art to control the timing of drug aerosolization relative to inhalation ( e . g ., breath - actuation ), to provide feedback to patients on the rate and / or volume of inhalation , to prevent excessive use ( i . e ., “ lock - out ” feature ), to prevent use by unauthorized individuals , and / or to record dosing histories . the dosage amount of an antidepressant in aerosol form is generally no greater than twice the standard dose of the drug given orally . the following are typical dosages of exemplary compounds for the treatment of depression : bupropion ( 100 mg ), nefazodone ( 150 mg ), perphenazine ( 2 mg ), trazodone ( 50 – 100 mg ), trimipramine ( 50 – 100 mg ), venlafaxine ( 75 mg ), tranylcypromine ( 15 mg ), citalopram ( 20 mg ), fluoxetine ( 20 mg ), fluvoxamine ( 50 mg ), mirtazepine ( 15 mg ), paroxetine ( 10 – 25 mg ), sertraline ( 25 – 50 mg ), amoxapine ( 25 – 250 mg ), clomipramine ( 100 mg ), doxepin ( 75 mg ), imipramine ( 75 mg ), maprotiline ( 50 mg ), nortriptyline ( 50 mg ), valproic acid ( 250 mg ) and protryptyline ( 15 mg ). as aerosols , the compounds are generally provided in the following amounts per inspiration for the same indication : bupropion ( 5 – 200 mg ), nefazodone ( 5 – 250 mg ), perphenazine ( 0 . 2 – 5 mg ), trazodone ( 5 – 200 mg ), trimipramine ( 5 – 200 mg ), venlafaxine ( 5 – 150 mg ), tranylcypromine ( 3 – 30 mg ), citalopram ( 4 – 40 mg ), fluoxetine ( 4 – 40 mg ), fluvoxamine ( 5 – 100 mg ), mirtazepine ( 3 – 30 mg ), paroxetine ( 2 – 50 mg ), sertraline ( 5 – 100 mg ), amoxapine ( 5 – 200 mg ), clomipramine ( 5 – 200 mg ), doxepin ( 5 – 150 mg ), imipramine ( 5 – 150 mg ), maprotiline ( 5 – 100 mg ), nortriptylene ( 5 – 100 mg ), valproic acid ( 20 – 1000 mg ) and protriptyline ( 3 – 30 mg ). a typical dosage of an antidepressant aerosol is either administered as a single inhalation or as a series of inhalations taken within an hour or less ( dosage equals sum of inhaled amounts ). where the drug is administered as a series of inhalations , a different amount may be delivered in each inhalation . one can determine the appropriate dose of an antidepressant containing aerosol to treat a particular condition using methods such as animal experiments and a dose - finding ( phase i / ii ) clinical trial . one animal experiment involves measuring plasma concentrations of drug in an animal after its exposure to the aerosol . mammals such as dogs or primates are typically used in such studies , since their respiratory systems are similar to that of a human . initial dose levels for testing in humans is generally less than or equal to the dose in the mammal model that resulted in plasma drug levels associated with a therapeutic effect in humans . dose escalation in humans is then performed , until either an optimal therapeutic response is obtained or a dose - limiting toxicity is encountered . analysis of antidepressant containing aerosols purity of an antidepressant containing aerosol is determined using a number of methods , examples of which are described in sekine et al ., journal of forensic science 32 : 1271 – 1280 ( 1987 ) and martin et al ., journal of analytic toxicology 13 : 158 – 162 ( 1989 ). one method involves forming the aerosol in a device through which a gas flow ( e . g ., air flow ) is maintained , generally at a rate between 0 . 4 and 60 l / min . the gas flow carries the aerosol into one or more traps . after isolation from the trap , the aerosol is subjected to an analytical technique , such as gas or liquid chromatography , that permits a determination of composition purity . a variety of different traps are used for aerosol collection . the following list contains examples of such traps : filters ; glass wool ; impingers ; solvent traps , such as dry ice - cooled ethanol , methanol , acetone and dichloromethane traps at various ph values ; syringes that sample the aerosol ; empty , low - pressure ( e . g ., vacuum ) containers into which the aerosol is drawn ; and , empty containers that fully surround and enclose the aerosol generating device . where a solid such as glass wool is used , it is typically extracted with a solvent such as ethanol . the solvent extract is subjected to analysis rather than the solid ( i . e ., glass wool ) itself . where a syringe or container is used , the container is similarly extracted with a solvent . the gas or liquid chromatograph discussed above contains a detection system ( i . e ., detector ). such detection systems are well known in the art and include , for example , flame ionization , photon absorption and mass spectrometry detectors . an advantage of a mass spectrometry detector is that it can be used to determine the structure of antidepressant degradation products . particle size distribution of an antidepressant containing aerosol is determined using any suitable method in the art ( e . g ., cascade impaction ). an andersen eight stage non - viable cascade impactor ( andersen instruments , smyrna , ga .) linked to a furnace tube by a mock throat ( usp throat , andersen instruments , smyrna , ga .) is one system used for cascade impaction studies . inhalable aerosol mass density is determined , for example , by delivering a drug - containing aerosol into a confined chamber via an inhalation device and measuring the mass collected in the chamber . typically , the aerosol is drawn into the chamber by having a pressure gradient between the device and the chamber , wherein the chamber is at lower pressure than the device . the volume of the chamber should approximate the tidal volume of an inhaling patient . inhalable aerosol drug mass density is determined , for example , by delivering a drug - containing aerosol into a confined chamber via an inhalation device and measuring the amount of active drug compound collected in the chamber . typically , the aerosol is drawn into the chamber by having a pressure gradient between the device and the chamber , wherein the chamber is at lower pressure than the device . the volume of the chamber should approximate the tidal volume of an inhaling patient . the amount of active drug compound collected in the chamber is determined by extracting the chamber , conducting chromatographic analysis of the extract and comparing the results of the chromatographic analysis to those of a standard containing known amounts of drug . inhalable aerosol particle density is determined , for example , by delivering aerosol phase drug into a confined chamber via an inhalation device and measuring the number of particles of given size collected in the chamber . the number of particles of a given size may be directly measured based on the light - scattering properties of the particles . alternatively , the number of particles of a given size is determined by measuring the mass of particles within the given size range and calculating the number of particles based on the mass as follows : total number of particles = sum ( from size range 1 to size range n ) of number of particles in each size range . number of particles in a given size range = mass in the size range / mass of a typical particle in the size range . mass of a typical particle in a given size range = π * d 3 * φ / 6 , where d is a typical particle diameter in the size range ( generally , the mean boundary mmads defining the size range ) in microns , φ is the particle density ( in g / ml ) and mass is given in units of picograms ( g − 2 ). rate of inhalable aerosol particle formation is determined , for example , by delivering aerosol phase drug into a confined chamber via an inhalation device . the delivery is for a set period of time ( e . g ., 3 s ), and the number of particles of a given size collected in the chamber is determined as outlined above . the rate of particle formation is equal to the number of 100 nm to 5 micron particles collected divided by the duration of the collection time . rate of aerosol formation is determined , for example , by delivering aerosol phase drug into a confined chamber via an inhalation device . the delivery is for a set period of time ( e . g ., 3 s ), and the mass of particulate matter collected is determined by weighing the confined chamber before and after the delivery of the particulate matter . the rate of aerosol formation is equal to the increase in mass in the chamber divided by the duration of the collection time . alternatively , where a change in mass of the delivery device or component thereof can only occur through release of the aerosol phase particulate matter , the mass of particulate matter may be equated with the mass lost from the device or component during the delivery of the aerosol . in this case , the rate of aerosol formation is equal to the decrease in mass of the device or component during the delivery event divided by the duration of the delivery event . rate of drug aerosol formation is determined , for example , by delivering an antidepressant containing aerosol into a confined chamber via an inhalation device over a set period of time ( e . g ., 3 s ). where the aerosol is pure antidepressant , the amount of drug collected in the chamber is measured as described above . the rate of drug aerosol formation is equal to the amount of antidepressant collected in the chamber divided by the duration of the collection time . where the antidepressant containing aerosol comprises a pharmaceutically acceptable excipient , multiplying the rate of aerosol formation by the percentage of antidepressant in the aerosol provides the rate of drug aerosol formation . the antidepressant containing aerosols of the present invention are typically used for the treatment of depression . valproic acid is also typically used for the treatment of mania . the following examples are meant to illustrate , rather than limit , the present invention . bupropion hydrochloride , perphenazine , trazodone hydrochloride , trimipramine maleate , tranylcypromine hydrochloride , citalopram hydrobromide , fluoxetine hydrochloride , fluvoxamine maleate , amoxapine , clomipramine hydrochloride , doxepin hydrochloride , imipramine hydrochloride , valproic acid , and maprotiline hydrochloride are commercially available from sigma ( www . sigma - aldrich . com ). nefazodone hydrochloride ( serzone ®), venlafaxine hydrochloride ( effexor ®), mirtazepine ( remeron ®), paroxetine hydrochloride ( paxil ®), sertraline hydrochloride ( zoloft ®), nortriptyline hydrochloride ( mylan ), and protriptylene hydrochloride ( vivactil ®) are commercially available , and the active ingredient can be isolated using standard methods in the art . approximately 1 g of salt ( e . g ., mono hydrochloride ) is dissolved in deionized water (˜ 30 ml ). three equivalents of sodium hydroxide ( 1 n naoh aq ) is added dropwise to the solution , and the ph is checked to ensure it is basic . the aqueous solution is extracted four times with dichloromethane (˜ 50 ml ), and the extracts are combined , dried ( na 2 so 4 ) and filtered . the filtered organic solution is concentrated using a rotary evaporator to provide the desired free base . if necessary , purification of the free base is performed using standard methods such as chromatography or recrystallization . a solution of drug in approximately 120 μl dichloromethane is coated on a 3 . 5 cm × 7 . 5 cm piece of aluminum foil ( precleaned with acetone ). the dichloromethane is allowed to evaporate . the coated foil is wrapped around a 300 watt halogen tube ( feit electric company , pico rivera , calif . ), which is inserted into a glass tube sealed at one end with a rubber stopper . running 90 v of alternating current ( driven by line power controlled by a variac ) through the bulb for 3 . 5 s ( 2 mg coating ) or 5 s ( 10 mg coating ) affords thermal vapor ( including aerosol ), which is collected on the glass tube walls . ( when desired , the system is flushed through with argon prior to volatilization .) reverse - phase hplc analysis with detection by absorption of 225 nm light is used to determine the purity of the aerosol . table 1 , which follows , provides data from drugs volatilized using the above - recited general procedure . to obtain higher purity aerosols , one can coat a lesser amount of drug , yielding a thinner film to heat . a linear decrease in film thickness is associated with a linear decrease in impurities . a solution of 22 . 0 mg paroxetine in 200 μl dichloromethane was spread out in a thin layer on the central portion of a 3 . 5 cm × 7 cm sheet of aluminum foil . the dichloromethane was allowed to evaporate . assuming a drug density of about 1 g / cc , the calculated thickness of the paroxetine thin layer on the 24 . 5 cm 2 aluminum solid support , after solvent evaporation , is about 9 . 0 microns . the aluminum foil was wrapped around a 300 watt halogen tube , which was inserted into a t - shaped glass tube . both of the openings of the tube were sealed with parafilm , which was punctured with ten needles for flow . the third opening was connected to a 1 liter , 3 - neck glass flask . the glass flask was further connected to a large piston capable of drawing 1 . 1 liters of air through the flask . alternating current was run through the halogen bulb by application of 90 v using a variac connected to 110 v line power . within 1 s , an aerosol appeared and was drawn into the 1 l flask by use of the piston , with collection of the aerosol terminated after 6 s . the aerosol was analyzed by connecting the 1 l flask to an eight - stage andersen non - viable cascade impactor . results are shown in table 1 . mmad of the collected aerosol was 2 . 0 microns with a geometric standard deviation of 1 . 9 . also shown in table 1 is the number of particles collected on the various stages of the cascade impactor , given by the mass collected on the stage divided by the mass of a typical particle trapped on that stage . the mass of a single particle of diameter d is given by the volume of the particle , πd 3 / 6 , multiplied by the density of the drug ( taken to be 1 g / cm 3 ). the inhalable aerosol particle density is the sum of the numbers of particles collected on impactor stages 3 to 8 divided by the collection volume of 1 l , giving an inhalable aerosol particle density of 9 . 0 × 10 6 particles / ml . the rate of inhalable aerosol particle formation is the sum of the numbers of particles collected on impactor stages 3 through 8 divided by the formation time of 6 s , giving a rate of inhalable aerosol particle formation of 1 . 5 × 10 9 particles / second . a solution of 19 . 6 mg paroxetine in 200 μl dichloromethane was spread out in a thin layer on the central portion of a 3 . 5 cm × 7 cm sheet of aluminum foil . the dichloromethane was allowed to evaporate . assumimz a drug density of about 1 g / cc , the calculated thickness of the paroxetine thin layer on the 24 . 5 cm 2 aluminum solid support , after solvent evaporation , is about 8 . 0 microns . the aluminum foil was wrapped around a 300 watt halogen tube , which was inserted into a t - shaped glass tube . both of the openings of the tube were sealed with parafilm , which was punctured with ten needles for flow . the third opening was connected to a 1 liter , 3 - neck glass flask . the glass flask was further connected to a large piston capable of drawing 1 . 1 liters of air through the flask . alternating current was run through the halogen bulb by application of 90 v using a variac connected to 110 v line power . within seconds , an aerosol appeared and was drawn into the 1 l flask by use of the piston , with formation of the aerosol terminated after 6 s . the aerosol was allowed to sediment onto the walls of the 1 l flask for approximately 30 minutes . the flask was then extracted with acetonitrile and the extract analyzed by hplc with detection by light absorption at 225 nm . comparison with standards containing known hplc with detection by light absorption at 225 nm . comparison with standards containing known amounts of paroxetine revealed that 3 . 4 mg of & gt ; 88 % pure paroxetine had been collected in the flask , resulting in an aerosol drug mass density of 3 . 4 mg / l . the aluminum foil upon which the paroxetine had previously been coated was weighed following the experiment . of the 19 . 6 mg originally coated on the aluminum , 7 . 4 mg of the material was found to have aerosolized in the 6 s time period , implying a rate of drug aerosol formation of 1 . 2 mg / s . a solution of 18 . 7 mg mirtazepine in 200 μl dichloromethane was spread out in a thin layer on the central portion of a 3 . 5 cm × 7 cm sheet of aluminum foil . the dichloromethane was allowed to evaporate . assuming a drug density of about 1 g / cc , the calculated thickness of the mirtazepine thin layer on the 24 . 5 cm 2 aluminum solid support , after solvent evaporation , is about 7 . 6 microns . the aluminum foil was wrapped around a 300 watt halogen tube , which was inserted into a t - shaped glass tube . both of the openings of the tube were sealed with parafilm , which was punctured with ten needles for air flow . the third opening was connected to a 1 liter , 3 - neck glass flask . the glass flask was further connected to a large piston capable of drawing 1 . 1 liters of air through the flask . alternating current was run through the halogen bulb by application of 90 v using a variac connected to 110 v line power . within 1 s , an aerosol appeared and was drawn into the 1 l flask by use of the piston , with collection of the aerosol terminated after 6 s . the aerosol was analyzed by connecting the 1 l flask to an eight - stage andersen non - viable cascade impactor . results are shown in table 1 . mmad of the collected aerosol was 1 . 2 microns with a geometric standard deviation of 2 . 2 . also shown in table 1 is the number of particles collected on the various stages of the cascade impactor , given by the mass collected on the stage divided by the mass of a typical particle trapped on that stage . the mass of a single particle of diameter d is given by the volume of the particle , πd 3 / 6 , multiplied by the density of the drug ( taken to be 1 g / cm 3 ). the inhalable aerosol particle density is the sum of the numbers of particles collected on impactor stages 3 to 8 divided by the collection volume of 1 l , giving an inhalable aerosol particle density of 5 . 6 × 10 7 particles / ml . the rate of inhalable aerosol particle formation is the sum of the numbers of particles collected on impactor stages 3 through 8 divided by the formation time of 6 s , giving a rate of inhalable aerosol particle formation of 9 . 3 × 10 6 particles / second . a solution of 20 . 7 mg mirtazepine in 200 μl dichioromethane was spread out in a thin layer on the central portion of a 3 . 5 cm × 7 cm sheet of aluminum foil . the dichloromethane was allowed to evaporate . assuming a drug density of about 1 g / cc . the calculated thickness of the mirtazepine thin layer on the 24 . 5 cm 2 aluminum solid support , after solvent evaporation , is about 8 . 4 microns . the aluminum foil was wrapped around a 300 watt halogen tube , which was inserted into a t - shaped glass tube . both of the openings of the tube were sealed with paraflim , which was punctured with ten needles for air flow . the third opening was connected to a 1 liter , 3 - neck glass flask . the glass flask was further connected to a large piston capable of drawing 1 . 1 liters of air through the flask . alternating current was run through the halogen bulb by application of 90 v using a variac connected to 110 v line power . within seconds , an aerosol appeared and was drawn into the 1 l flask by use of the piston , with formation of the aerosol terminated after 6 s . the aerosol was allowed to sediment onto the walls of the 1 l flask for approximately 30 minutes . the flask was then extracted with acetonitrile and the extract analyzed by hplc with detection by light absorption at 225 nm . comparison with standards containing known amounts of mirtazepine revealed that 10 . 65 mg of & gt ; 99 % pure mirtazepine had been collected in the flask , resulting in an aerosol drug mass density of 10 . 65 mg / l . the aluminum foil upon which the mirtazepine had previously been coated was weighed following the experiment . of the 20 . 7 mg originally coated on the aluminum , 18 . 7 mg of the material was found to have aerosolized in the 6 s time period , implying a rate of drug aerosol formation of 3 . 1 mg / s . valproic acid (˜ 90 mg ) was adsorbed onto a piece of glass wool . the coated glass wool was inserted into a glass tube in a furnace ( tube furnace ). a glass wool plug was placed in the tube adjacent to the foil sheet , and an air flow of 2 l / min was applied . the furnace was heated to 300 ° c . for 120 s to volatilize the coated valproic acid and then was allowed to cool . the glass wool was extracted , and hplc analysis of the collected material showed it to be at least 99 . 5 % pure valproic acid .

the present invention relates to the delivery of antidepressants through an inhalation route . specifically , it relates to aerosols containing an antidepressant that are used in inhalation therapy . in a method aspect of the present invention , an antidepressant is administered to a patient through an inhalation route . the method comprises : a ) heating a thin layer of an antidepressant , on a solid support , form a vapor ; and , b ) passing air through the heated vapor to produce aerosol particles having less than 5 % antidepressant degradation products . in a kit aspect of the present invention , a kit for delivering an antidepressant through an inhalation route to a mammal is provided which comprises : a ) a thin coating of an antidepressant composition and b ) a device for dispensing said thin coating as a condensation aerosol .

the therapeutic laser can be designed as a cw laser or as a repeatedly pulsed laser so that optoacoustic measurements can be carried out at all , pulsed electromagnetic radiation must of necessity be applied to the retina , preferably precisely to the laser spot treated by the therapeutic laser . these radiation pulses per se only have to provide for rather small temperature variations ( less than 1 ° c . ), to produce pressure waves that can be measured . therefore they can be irradiated from a second probe laser that is completely independent from the therapeutic laser ( cf . de 101 35 944 c2 ), both beams being preferably mirrored into the same beam path . in actual fact , using a laser for producing the optoacoustic signal is in no way imperative . a broad - band light source with pulsed operation ( for example sld , superluminescent diode ) or a repeating flashlight can be used as well ( in principle even modulated microwaves or x - rays were possible , but preferably not on the eye ). a precise delimitation of the pulsed excitation radiation to the therapeutic laser spot is favorable , but not at all necessary . a repeatedly pulsed therapeutic laser ( e . g . pulse power 100 watts , pulse duration around 100 ns , repetition rate about 10 khz , on / off ratio approximately 1 : 1000 ) is however on its own sufficient for realizing the invention . the effect of such a “ nanopulsing ” laser on biological tissue can be compared to that of cw irradiation , but each individual pulse produces a short - term temperature increase in the tissue by fractions of one degree centigrade . this entails pressure waves due to the material expanding , that can likewise be detected using the pressure sensor . in the following text it is always assumed that the therapeutic laser has been designed to apply repeatedly pulsed laser light onto the retina independently of the fact whether or not it is supplied with it from an additional probe laser light source . even a cw laser with a chopper can be considered . for simplification reasons , other designs of the therapeutic laser , for example using flashlights etc ., are not discussed any further . that the functioning described further below can be transferred to such designs will be readily obvious to the person skilled in the art . the control unit comprises all means known per se to change the beam of the therapeutic laser , that is to say in particular means for checking the pump power or even an acousto - optic modulator that deflects a proportion of the emitted laser light from the beam path toward the retina , and drivable beam optics that can in particular widen the beam . laser power , beam diameter , and irradiation time are the parameters to be regulated , that control the tissue damage to be achieved . the detection device is preferably designed as an ultrasound converter that is arranged on the requisite contact lens and reacts to pressure fluctuations by generating electrical signals . the signals are proportional to the pressure amplitude and are fed to the evaluation device . it shall be remarked at this point that there are still other possible embodiments of the detection device , in particular those for non - contacting measurements . they are not covered here in more detail , but the present invention should not be regarded as being limited to a specific type of measurement of the pressure transients . the new aspect of the inventive apparatus is in the type of evaluation of the recorded signals by the evaluation device . the latter has two tasks to fulfill : 1 . the pressure transients are sampled by the detection device as functions of the time and transmitted to the evaluation device . the evaluation device continually measures the time that has elapsed , as long as the therapeutic laser irradiates the retina . for each pressure transient that is caused by a single light pulse it calculates an auxiliary parameter that is to be referred to here as an evaluation measure . the maximum amplitude or the surface area below the pressure transient curve ( for example absolute integral ) can be considered as the evaluation measure , but also phase shifts and frequency changes of the pressure transients inform on the state changes of the tissue due to the impact of the therapeutic radiation . it has been shown that these parameters on average correlate well with the temperature of the ocular fundus , e . g . pressure and phase changes are proportional to temperature changes over short distances . since a value of the evaluation measure of the pressure transients is formed for each light pulse , the evaluation measure then exists in a first interval of duration δt 1 as a function of time b ( t ). this first interval starts with the onset of the therapeutic radiation (= start of the time measurement ) and be referred to as “ startup phase ”. 2 . usually , b ( t ) is initially very noisy and is smoothed as required by the evaluation device by means of a smoothing procedure , for example by means of window averaging with a window width very much smaller than δt 1 . the smoothed function & lt ; b ( t )& gt ; is fitted at the end of the startup phase using a conventional procedure ( e . g . least squares or similar ) by a simple analytical function that estimates in advance the future course of the evaluation measure by extrapolation while maintaining current irradiation parameters . the evaluation device compares the fit parameters that have been found for the simple analytical function with data listed in tables in the internal data storage unit . on the basis of this comparison , the control unit is driven to match the irradiation parameters . in the following text , the functioning of the inventive apparatus is explained in more detail . after a few preliminary remarks , specific designs of the invention are illustrated . fig1 represents curves for the temperature - time dependency of the denaturation of proteins in the retina from the literature ( curves 1 and 2 ). both curves show isolines with an identical extent of the damage ( curve 1 : ω = 1 , coagulation that is just visible ; curve 2 : ω = 100 , strong coagulation ) as a function of the selected exposition parameters ( here : temperature and time ). fig1 also shows the temperature rise , calculated assuming identical absorption , when a 200 μm spot is irradiated with varying laser powers in the green spectral range at the center of the retinal pigment epithelium as the strongest absorber ( curve 3 : 20 mw , curve 4 : 30 mw , curve 5 : 40 mw ). the calculation does not take into account any change in the tissue . the ω family of curves ( 1 , 2 ) and the family of curves ( 3 , 4 , 5 ) obviously intersect sooner or later , and in particular the intersection points with curve 1 specify those times for the different laser powers when coagulation is first visible . to better illustrate this , fig2 shows the dependency of this time t coag on the laser power as a curve . with the same degree of absorption , different laser powers lead to different temperature rises shortly after switching on the therapeutic laser . as an alternative , fig3 therefore shows the time t coag as a function of the temperature increase ( curve 6 ). since the grüneisen coefficient can be approximated linearly very well over small temperature ranges , the change in the pressure amplitude δp is plotted on the abscissa in fig3 and can be measured directly . ( reminder : the pressure amplitude is a possible evaluation measure of the pressure transient .) converting into temperatures is thus not necessary . even if the second approximation ( parabolic development of the temperature with the pressure ) is considered , the times change only little up to coagulation ( curve 7 ). the pressure rise up to the start of the treatment is therefore a direct measure for the time when coagulation sets in . the speed of this increase cannot be predicted before the start of the irradiation if the material properties are not precisely known at the selected location of the laser spot ( this is always the case in particular for retina tissue ). from the course of the evaluation measure during the startup phase in which as yet no tissue changes occur , there results according to the invention a sensible estimation of the total time until coagulation , by extrapolation and determining intersection points with the previously known ω curves that describe the extent of the damage . the ω curves exist for example as value tables in the data storage unit of the evaluation device that carries out the fitting procedure and calculates the intersection of the fit curve with a selected ω curve . since this calculation takes place immediately after the initial phase , the result is the residual time until the desired tissue damage is reached . in a first embodiment of the invention , the evaluation device controls the control device after the pre - calculated residual time has expired and causes the therapeutic laser to be switched off . the treating physician can preferably reactivate the therapeutical laser only manually , generally after the change to another laser spot . the extent of the desired tissue damage can be selected by the physician prior to the start of the treatment by manually programming the evaluation device . preferably he selects a ω value , being guided by a menu . the first design of the invention proceeds from two basic requirements : 1 . a body of evidence has to exist on the course of the ω family of curves , for example from literature or from one &# 39 ; s own preliminary examinations that may possibly also be costly . here the arrhenius model discussed in the prior art does not represent the only possibility to define tissue damage . in particular it can be convenient for different clinical pictures or therapeutical approaches to determine totally different models or new value tables and to store them in the memory of the evaluation device . ultimately , prior to the therapeutical usage of the apparatus the family of curves must have been determined empirically and sufficiently precisely for the evaluation measure to be considered in each case . 2 . since the fitting procedure of the evaluation device is to serve for extrapolation of & lt ; b ( t )& gt ; after the starting phase , a reasonable fit function has to be pre - specified . a selection that makes sense physically is one that ensures that the fit function approaches a finite end value for large exposition times . what is being suggested specifically here , is the function f ( t )= a − b exp (− λ t ) with a , b , λ as fitting parameters . ( 1 ) it starts at t = 0 ( start of the therapeutic radiation ) with the starting value a − b , has an initial slope of bλ and slowly approaches the end value a . the fitting parameters are easy to determine from the values of & lt ; b ( t )& gt ; determined during the starting phase , so that as a good approximation & lt ; b ( t )≈ f ( t ) holds for 0 ≦ t ≦ δt 1 ( 2 ) the evaluation device now calculates the intersection of f ( t ) with the preselected ω curve from the data storage unit and calculates the time t coag & gt ; δt 1 when the therapeutic laser is to be switched off . this time obviously depends on the choice of the fit function f ( t ) that could well also have another shape . for example , also a polynomial could be considered . here again , it may be favorable to use different fit functions for different therapeutical purposes and / or clinical images . even if the inventive apparatus , presented up till now , is used , there remains a certain degree of uncertainty as to the tissue damage that has been actually achieved . however , this uncertainty is considerably reduced relative to the measures according to the state of the art — visual inspection by the physician . a further improvement can be achieved with a second design of the invention . according to what has been said previously , an optimum fit function f ( t ) can be determined unanimously at most in the individual case . when it is selected , it is still possible to over - or underestimate systematically the true development of the evaluation measure , so that the therapeutic radiation is then always applied for too short or too long a time . this entails that the tissue damage can be particularly different between two laser spots that differ strongly in terms of their absorption — and thus in their initial slope of the evaluation measure . according to the invention , a repeatable measurement and evaluation process is additionally provided . for the fitting parameters determined during the initial phase it is checked whether the course of f ( t ) corresponds to a predetermined positive criterion . if yes , lasering is continued as described above , otherwise the laser power is changed . the positive criterion is to be in particular that during the temporal range of the initial phase δt 1 the calculated fit function is completely inside a predetermined corridor , that is to say only a limited selection of initial courses is permitted . in particular the initial slopes ( e . g . according to equation ( 1 ): wo is limited to a specific range of values , but also the curvature behavior of f ( t ) can be restricted in such a way . the restriction makes sense in particular so as to keep the irradiation times within certain limits . thus it is known for example that irradiation times that are too long ( several 100 ms ) entail the risk of involuntary eye movements of the patient . in particular during the laser therapy in the area of the macula , this is what you want to avoid . if , however , the inventive measurement of the initial slope of the evaluation measure suggests that only a very short irradiation time ( e . g . & lt ; 30 ms ) would be necessary for coagulation , then due to the then very rapid heating - up and the start of the vaporization the risk of retinal bleeding would exist , as is well documented in the literature for pulse durations of a few milliseconds . if during the initial phase the calculated fit function does not keep to the corridor that is preferably stored for comparison reasons in the data memory , for example in the form of two limit functions f min ( t ) and f max ( t ) between which f ( t ) may move , the therapeutic laser is preferably deactivated until the tissue has cooled down again to its starting temperature . since the initial phase is only short , this happens very rapidly and the tissue has not yet suffered any damage . the evaluation device commands the control device to increase the power of the therapeutic laser if the fit function leaves the corridor in the downward direction , or otherwise to reduce it . the extent of the power matching should preferably be geared to the extent of the deviation between fit function and corridor . the most simple , convenient matching consists in multiplying the laser power with the ratio of the average desired slope of the evaluation measure ( which is indeed prescribed ) and the slope actually measured at the laser spot . it is also the radiation intensity ( power / surface area ) that can be changed instead of the laser power by appropriately widening or narrowing the beam . if the therapeutic laser is ultimately reactivated , the measurement and evaluation process above starts from scratch at the same laser spot with the power that has now been changed . the matching steps are repeated until the positive criterion is finally fulfilled . reactivation of the therapeutic laser has to take place fully automatically between the different matching procedures of the laser power — in contrast to the manual activation mentioned above during a transition from one laser spot to the next one . the apparatus can set different powers within fractions of a second and “ test ” it on the retina . the treating physician does not see the automatic process and all the while keeps the applicator pointed only at the therapeutical location selected by him . if he had to activate the therapeutic laser himself , he would most likely shift the laser spot due to the movement and matching would possibly never succeed . a special case of the previous design with iterative matching of the laser intensity ( power and / or irradiation surface ) is the one - step iteration that should get a special mention . here , if a deviation of the starting course of the evaluation measure from the predetermined corridor is established , no deactivation of the therapeutic laser is carried out . the power of the therapeutic laser is instead matched during current operation — as described by multiplication with a ratio value . the time t coag until the onset of the coagulation is not calculated separately here , but results from the pre - known course of the desired curve , e . g . in the center of the corridor , i . e . it is predetermined . this special design of the invention is certainly somewhat less precise than the above - described apparatus with iteration , because it dispenses with a check of the success of the control measure and its optimization . however , it has the advantage that it guarantees that the treatment duration of a single laser spot corresponds to that which is common nowadays . in contrast thereto , the iteratively operating apparatus safely provides for an approximation of the evaluation measure to the desired course or the corridor . in practice , the absorptivity present at the current laser spot is measured , and fully - automatically matching the laser power leads to the energy deposition taking place in all laser spots in the same way , in particular with the same temporal course . herein is seen the presently best - possible guarantee to effect a constant damage during the entire treatment .

a therapeutic laser with a source of pulsed electromagnetic radiation , a control device for controlling the intensity and / or the duration of the therapeutic laser applied to the tissue , and a detection device for detecting optoacoustic signals triggered by irradiating the living tissue with the pulsed electromagnetic radiation . the therapeutic laser is characterized by an evaluation device that acts on the control device and is used for calculating a degree of quality b from the optoacoustic signals detected by the detection device for individual laser pulses applied to a predetermined laser spot and determining a fit function f at a predetermined point in time δt1 , the fit function f approximating the mean curve of b for 0 ≦ t ≦ δt1 . the intensity and / or the irradiation time of the therapeutic laser is defined by the parameters for the predetermined laser spot , the parameters being determined for the fit function f .

the present inventors identified the molecular events linking fat4 and amotl1 to cardiac growth , and showed that fat4 is required to restrict cardiomyocyte hypertrophy and cardiomyocyte proliferation , and that this restriction involves two activators of the hippo signalling pathway , namely amotl1 and yap1 . more precisely , they show that fat4 is required to organise cell junctions and sequester amotl1 , preventing excessive heart growth . in the absence of fat4 , amotl1 is released and , in a complex with yap1 , translocates to the nucleus , bypassing the hippo kinases . resulting variations in gene expression promote proliferation and hypertrophy of cardiomyocytes , leading to excessive growth of the myocardium . treating methods , diagnosis methods as well as screening methods can be contemplated in light of these new findings . the present invention proposes to use fat4 - dependent hippo pathway modulators in cardiac repair . fat4 - dependent hippo pathway modulators are for example amotl1 or yap1 , which have been shown to activate cardiac cell hypertrophy and regeneration , or fat4 itself , which conversely restricts heart growth ( see experimental part below ). more precisely , the results of the present inventors highlight that it is possible to : i ) prevent or reduce heart growth , heart regeneration , and / or cardiomyocyte proliferation by down - regulating the expression of fat4 dependent hippo pathway activators , namely yap1 or amotl1 , or by up - regulating the expression of fat4 in cardiomyocytes , ii ) reactivate cardiomyocyte proliferation or enhance heart size by down - regulating the expression of fat4 or by up - regulating the expression of fat4 dependent hippo pathway activators , namely yap1 or amotl1 in cardiomyocytes , or by targeting amotl1 to the nucleus or by preventing the sequestration of amotl1 at cell junction or in a complex with fat4 . fat4 ( or fat atypical cadherin 4 or protocadherin fat4 ) is encoded by the fat4 cdna of seq id no : 1 in mouse ( nm_183221 . 3 ), seq id no : 2 in human ( nm_001291303 . 1 ) and seq id no : 3 in rat ( nm_001191705 . 1 ). the encoded polypeptide is a member of the protocadherin family , involved in planar cell polarity . these cdna encode the fat4 polypeptide of seq id no : 4 ( mouse fat4 , np_899044 . 3 ), seq id no : 5 ( human fat4 , np_001278232 . 1 ) and seq id no : 6 ( rat fat4 , np_001178634 . 1 ), respectively . yap1 ( or yes - associated protein 1 , also known as yap65 ) is encoded by the yap1 cdna of seq id no : 7 in mouse ( nm_001171147 . 1 ), seq id no : 8 in human ( nm_001130145 . 2 ) and seq id no : 9 in rat ( nm_001034002 . 2 ). the yap1 gene is known to play a role in the development and progression of multiple cancers as a transcriptional regulator of this signaling pathway and may function as a potential target for cancer treatment . it encodes the yap1 polypeptide of seq id no : 10 ( mouse yap1 , np_001164618 . 1 ), seq id no : 11 ( human yap1 , np_001123617 . 1 ) and seq id no : 12 ( rat yap1 , np_001029174 . 2 ), respectively . angiomotin - like protein 1 ( amotl1 ) is a peripheral membrane protein that is a component of tight junctions ( tjs ). tjs form an apical junctional structure and act to control paracellular permeability and maintain cell polarity . this protein is related to angiomotin , an angiostatin binding protein that regulates endothelial cell migration and capillary formation ( nishimura m , kakizaki m , ono y , morimoto k , takeuchi m , inoue y , imai t , takai y ( february 2002 ). “ jeap , a novel component of tight junctions in exocrine cells ”. j biol chem 277 ( 7 ): 5583 - 7 ). it is encoded by the amotl1 cdna having the seq id no : 13 ( nm_001081395 . 1 ) in mouse , seq id no : 14 in human ( nm_130847 . 2 ), and seq id no : 15 ( xm_008766026 . 1 ) in rat . these cdnas encode the amotl1 polypeptide of seq id no : 16 ( mouse amotl1 , np_001074864 . 1 ), seq id no : 17 ( human amotl1 , np_570899 . 1 ) and seq id no : 18 ( rat amotl1 , xp_008764248 . 1 ), respectively . in a first aspect , the present invention therefore relates to a method for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal , comprising down - regulating the fat4 - dependent activator of the hippo pathway yap1 and / or amotl1 or up - regulating fat4 in said mammal . cardiomyocyte hyperproliferation induces an increase of the heart size that is usually designated as “ cardiac hypertrophy ” or “ mitogenic cardiomyopathy ”. thus , as used herein , the terms “ cardiac hypertrophy ” and “ mitogenic cardiomyopathy ” are equivalent . in a particular embodiment , said method comprises the step of down - regulating yap1 expression or transcriptional activity in said mammal , more particularly in the cardiomyocytes of said mammal . said down - regulation may be carried out by administering an effective amount of an anti - sense nucleotide inhibiting specifically yap1 gene expression . said anti - sense nucleotide is for example a sirna ( or dsrna ), a mirna , a shrna , a ddrnai . nuclease - based technologies such as zn - finger nuclease , tale nuclease or cas9 / crispr systems can also be used to inhibit gene expression . more specifically , these anti - sense nucleotides have approximately 15 to 30 nucleotides , 19 to 25 nucleotides , or preferably around 19 nucleotides in length . they are for example complementary ( strand 1 ) and identical ( strand 2 ) to a fragment of seq id no : 7 , seq id no : 8 or seq id no : 9 . sirnas are described for example in wo 02 / 44 321 ( mit / max planck institute ). this application describes a double strand rna ( or oligonucleotides of same type , chemically synthesized ) of which each strand has a length of 19 to 25 nucleotides and is capable of specifically inhibiting the post - transcriptional expression of a target gene via an rna interference process in order to determine the function of a gene and to modulate this function in a cell or body . also , wo 00 / 44895 ( biopharma ) concerns a method for inhibiting the expression of a given target gene in a eukaryote cell in vitro , in which a dsrna formed of two separate single strand rnas is inserted into the cell , one strand of the dsrna having a region complementary to the target gene , characterized in that the complementary region has at least 25 successive pairs of nucleotides . persons skilled in the art may refer to the teaching contained in these documents to prepare the sirnas of the invention . micrornas ( hereafter referred to as mirnas ) are small non - coding rna molecule ( ca . 22 nucleotides ) found in plants and animals , which functions in transcriptional and post - transcriptional regulation of gene expression . mirnas function via base - pairing with complementary sequences within mrna molecules , usually resulting in gene silencing via translational repression or target degradation . the skilled person may also use ddrnai molecules such as those described generic fashion in application wo 01 / 70949 ( benitec ). designing anti - sense nucleotides that are efficient in down - regulating yap1 expression in the targeted cells is well - known in the art . http :// sonnhammer . cgb . ki . se / sisearch / sisearch_1 . 6 . html ( improved and automated prediction of effective sirna ”, chaml a m , wahlesdelt c and sonnhammer e l l , biochemical and biophysical research communications , 2004 ). http :// design . mai . jp / sidirect / index . php ( direct : highly effective , target - specific sirna design software for mammalian rna interference , yuki naito et al , nucleic acids res , vol . 32 , n ° web server issue © oxford university press , 2004 ). “ sirna target finder ” by ambion at the address http :// www . ambion . com / techlib / misc / sirna_tools . html “ sirna design tool ” by whitehead institute of biomedical research at the mit at the address http :// jura . wi . mit . edu / pubint / http :// iona . wi . mit . edu / sirnaext / examples of anti - yap1 sirnas that can be used in the methods of the invention are provided in the enclosed listing sequence , as seq id no : 24 , seq id no : 25 and seq id no : 27 , that are specific of rat yap1 . in a particularly preferred embodiment , the present invention relates to an anti - sense nucleotide ( e . g ., a sirna ) inhibiting specifically the expression of yap1 , for use for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . in another particularly preferred embodiment , the present invention relates to the use of an anti - sense nucleotide ( e . g ., a sirna ) inhibiting specifically the expression of yap1 , in the manufacture of a medicament that is useful for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . by “ inhibiting specifically ”, it is herein meant compounds having an ic50 on the yap1 protein expression of less than 1 μm , preferably 100 nm , whereas it has an ic50 on any other protein of more than 5 μm or 10 μm . said down - regulation may also be carried out by administering an effective amount of a chemical compound that inhibits yap1 transcriptional activity . such compound is for example verteporfin or cardiac glycoside digitonin 44 . alternatively , down - regulation may also be carried out by administering an effective amount of a chemical compound that inhibits yap1 expression . in a particularly preferred embodiment , the present invention relates to verteporfin for use for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . in another particularly preferred embodiment , the present invention relates to the use of verteporfin , in the manufacture of a medicament that are useful for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . in another particular embodiment , said method comprises the step of down - regulating amotl1 expression or biological activity in said mammal , more particularly in the cardiomyocytes of said mammal . as shown in the experimental part below , amotl1 biological activity is dependent on its translocation to the nucleus , where it transports the transcription co - factor yap1 in the absence of fat4 ( in the presence of fat4 , amotl1 is sequestered at cell junctions in a complex involving fat4 ). thus , dowregulating amotl1 biological activity may be achieved by favoring the interaction of amotl1 and fat4 , or of amotl1 to cell junctions , thereby leading to its sequestration out of the nucleus . it is possible to assess this biological activity directly by detecting the subcellular localisation of amotl1 , e . g ., by immunohistochemistry or any conventional means , or indirectly by measuring the expression of amotl1 - dependent genes ( e . g . , aurkb , ccna2 , birc2 , birc5 , cdkn1b , lyh6 , or acta1 ). thus , down - regulation of amotl1 biological activity may be carried out by administering inhibitors ( e . g ., peptides ) of amotl1 - fat4 interaction or of amotl1 - yap1 interaction , or any compounds ( either chemical or peptides ) that would sequester amotl1 out of the cardiomyocyte nucleus . in this aim , it would be possible to use for example cardiac glycoside digitonin 44 . preferably , said down - regulation is carried out by administering an effective amount of an anti - sense nucleotide inhibiting specifically amotl1 gene expression . said anti - sense nucleotide is for example a sirna ( or dsrna ), a mirna , a shrna , a ddrnai . nuclease - based technologies such as zn - finger nuclease , tale nuclease or cas9 / crispr systems can also be used to inhibit gene expression . designing anti - sense nucleotides that are efficient in down - regulating amotl1 expression in the targeted cells is well - known in the art . these anti - sense nucleotides have preferably 15 to 30 nucleotides , 19 to 25 nucleotides , or more preferably around 19 nucleotides in length . they are for example complementary ( strand 1 ) and identical ( strand 2 ) to a fragment of seq id no : 13 , seq id no : 14 or seq id no : 15 . examples of anti - amotl1 sirnas that can be used in the methods of the invention are provided in the enclosed listing sequence , as seq id no : 21 to 23 , that are specific of rat amotl1 . in a particularly preferred embodiment , the present invention relates to an anti - sense nucleotide ( e . g ., a sirna ) inhibiting specifically the expression of amotl1 , for use for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . in another particularly preferred embodiment , the present invention relates to a compound inhibiting the nuclear translocation of amotl1or increasing the sequestration of amotl1 out of the nucleus , for use for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . in another particularly preferred embodiment , the present invention relates to the use of an anti - sense nucleotide ( e . g ., a sirna ) inhibiting specifically the expression of amotl1 , in the manufacture of a medicament that is useful for preventing and / or treating cardiac hypertrophy by reducing heart growth in a mammal . the results of the inventors show that overexpression of an intracellular domain of fat4 prevents the nuclear localisation of yap1 and therefore reduces cardiomyocyte proliferation ( fig3 ). in another particular embodiment , said method comprises the step of up - regulating fat4 expression or biological activity in said mammal , more particularly in the cardiomyocytes of said mammal . as shown in the experimental part below , fat4 biological activity in cardiomyocytes is based on the sequestration of amotl1 at cell junctions , i . e ., out of the nucleus where amotl1 may induce transcription of many proliferation genes . thus , upregulating fat4 biological activity may be achieved by favouring the interaction of amotl1 and fat4 , thereby leading to the sequestration of amotl1 out of the nucleus . it is possible to assess this biological activity directly by detecting the colocalisation of amotl1 with fat4 , e . g ., by immunohistochemistry ( or any other conventional means ), or indirectly by detecting the subcellular localisation of amotl1 in cardiomyocytes or by measuring the expression of amotl1 - dependent genes ( e . g ., aurkb , ccna2 , birc2 , birc5 , cdkn1b , lyh6 , or acta1 ). preferably , said up - regulation is achieved by administering a gene therapy vector encoding the fat4 polypeptide or a fragment of the fat4 polypeptide or by administering any compound activating the expression of the fat4 polypeptide . this vector is for example a viral vector encoding a fragment of the fat4 polypeptide . more precisely , this vector can be an aav vector ( e . g ., an aav9 vector , which has a good affinity for cardiomyocytes ) encoding fat4 or a fragment of the fat4 polypeptide . in a preferred embodiment , said mammal is a human . preferably , said human suffers from cardiac hypertrophy , as defined above . in another preferred embodiment , said mammal is embryonic or newborn . if it is newborn , it is more preferably one month or less of age , one week or less of age , or one day or less of age . in a particular embodiment , the present invention relates to a method for reducing heart growth in a mammal , comprising downregulating yap1 or upregulating fat4 in the mammal sufficient to restrict heart growth in the mammal , wherein the mammal is embryonic or newborn . the growth of the mammalian heart is critical for its contractile function . during development , cardiomyocyte proliferation underlies most of the growth , whereas increase in cell size ( hypertrophy ) predominates after birth ( li et al ., 1996 ). although resident stem cells of cardiomyocytes have been detected in the adult heart ( beltrami et al ., 2003 ; hsieh et al . 2007 ), their number and contribution to heart regeneration remains anecdotal . by clonal analysis , it was shown that growth of the embryonic myocardium follows an exponential mode of growth , indicating that symmetrical divisions of myocardial precursors or cardiomyocytes underlie heart growth , and that there is no major pool of cardiac stem cells ( meilhac et al ., 2003 ). 1 week after birth in the mouse and about 10 years in human ( li et al ., 1996 ; bergmann 2009 ), cardiomyocytes lose their potential of proliferation . this has been shown to be directly associated with the loss of the regeneration potential of the heart ( porrello et al ., 2011 ). however , adult cardiomyocytes retain some potential of proliferation ( bergmann et al ., 2009 ; senyo et al ., 2013 ; villa del campo et al ., 2014 ). thus , enhancing cardiomyocyte proliferation in situ in the more mature heart by exploiting the developmental pathways controlling heart growth seems particularly attractive as an approach for cardiac repair . in another aspect , the present invention relates to a method to induce heart growth in a mammal , comprising down - regulating fat4 in said mammal . in particular , said method comprises the down - regulation of fat4 in the cardiomyocytes of said mammal . in another particular embodiment , said method comprises the step of down - regulating fat4 expression or biological activity in said mammal , more particularly in the cardiomyocytes of said mammal . downregulating fat4 biological activity may be achieved by impairing the interaction of amotl1 and fat4 , thereby leading to the liberation of amotl1 and its translocation in the nucleus . it is possible to assess this biological activity directly by detecting the colocalisation of amotl1 with fat4 , e . g ., by immunohistochemistry ( or any other conventional means ), or indirectly by detecting the subcellular localisation of amotl1 in cardiomyocytes or by measuring the expression of amotl1 - dependent genes ( e . g ., aurkb , ccna2 , birc2 , birc5 , cdkn1b , lyh6 , or acta1 ). alternatively , down - regulating fat4 expression can be carried out by administering an effective amount of an anti - sense nucleotide inhibiting specifically fat4 gene expression . said anti - sense nucleotide is for example a sirna ( or dsrna ), a mirna , a shrna , a ddrnai . nuclease - based technologies such as zn - finger nuclease , tale nuclease or cas9 / crispr systems can also be used to inhibit gene expression . designing anti - sense nucleotides that are efficient in down - regulating fat4 expression in the targeted cells is well - known in the art . these anti - sense nucleotides have preferably 15 to 30 nucleotides , 19 to 25 nucleotides , or more preferably around 19 nucleotides in length . they are for example complementary ( strand 1 ) and identical ( strand 2 ) to a fragment of seq id no : 1 , seq id no : 2 or seq id no : 3 . examples of sirnas that can be used with this respect are provided in the enclosed listing sequence , as seq id no : 19 , seq id no : 20 and seq id no : 26 , that are specific of rat fat4 . in a particularly preferred embodiment , the present invention relates to an anti - sense nucleotide ( e . g ., a sirna ) inhibiting specifically the expression of fat4 , for use for inducing heart growth in a mammal or for amplifying a population of cardiomyocytes . in another particularly preferred embodiment , the present invention relates to the use of an anti - sense nucleotide ( e . g ., a sirna ) inhibiting specifically the expression of fat4 , in the manufacture of a medicament that is useful for inducing heart growth in a mammal . the above - mentioned anti - sense nucleotides can be injected into the cells or tissues by lipofection , transduction or electroporation or viral infection ( e . g ., by using an aav9 vector ). they can be used to specifically destroy the mrnas encoding yap1 , fat4 or amotl1 thereby entailing the possible therapeutic applications mentioned above . enhancing cardiomyocyte proliferation in vitro by exploiting the developmental pathways controlling cardiomyocyte proliferation is also particularly attractive for producing cardiac tissues that could be grafted in a patient . in another aspect , the present invention relates to an in vitro method for producing high amounts of cardiomyocytes , said method involving the upregulation of amotl1 or yap1 in the nucleus of said cells or the down - regulation of fat4 in the cytoplasm of said cells . b ) contacting said cardiomyocytes with a compound down - regulating fat4 , or with a compound upregulating nuclear amotl1 and / or yap1 , upregulation of amotl1 or of yap1 can be performed for example by transfecting cardiomyocytes with a vector encoding the amotl1 or the yap1 polypeptide . said vector preferably contains a nuclear localisation signal , so that the encoded polypeptide is forced to translocate to the nucleus of the transfected cells . in a preferred embodiment , said vector is an adenovirus . adequate vectors are disclosed in the experimental part below ( nlsamotl1 ). downregulation of fat4 can be performed by any of the above - mentioned means . the in vitro method of the invention can be carried out on primary cardiomyocyte cells that have been extracted from a cardiac tissue ( after a biopsy or cardiac surgery , for example ). in a more preferred embodiment , cardiomyocytes are generated by transforming stem cells ( either embryonic stem cells or ips cells ) into cardiomyocytes by a conventional mean ( goumans m . j . et al , stem cell res . 2007 ; laflamme m . a . et al , nat . biotechnol . 2007 ; van laake et al , stem cell res . 2007 ; blin g . et al , the journal of clinical investigation , 2010 ; blin et al , curr stem cell res ther 2010 ; christine l . et al , science translational medicine , 2010 ). in another aspect , the present invention relates to an in vitro method for diagnosing cardiac hypertrophy in a mammal , comprising analyzing the expression level of fat4 or amotl1 or yap1 or detecting inactivating mutations in the polypeptide sequence of fat4 , yap1 or amotl1 , in a tissue sample from said mammal . preferably , if fat4 expression level is reduced as compared with a reference value , or if the fat4 polypeptide contains at least one inactivating mutation , then said mammal is suffering from or will develop cardiac hypertrophy . preferably , if yap1 expression level is enhanced as compared with a reference value , then said mammal is suffering from or will develop cardiac hypertrophy . preferably , if amotl1 expression level is enhanced as compared with a reference value , then said mammal is suffering from or will develop cardiac hypertrophy . detection of reduced fat4 , yap1 or amotl1 expression level may be achieved by any conventional means ( qpcr , elisa , immunohistochemistry , etc .). the term “ reference value ”, as used herein , refers to the expression level of the fat4 , yap1 or amotl1 gene in a reference sample . a “ reference sample ”, as used herein , means a sample obtained from subjects , preferably two or more subjects , known not to suffer from cardiac hypertrophy . the suitable reference expression levels of fat4 , yap1 or amotl1 can be determined by measuring the expression levels of fat4 , yap1 or amotl1 in several suitable subjects , and such reference levels can be adjusted to specific subject populations . the reference value or reference level can be an absolute value ; a relative value ; a value that has an upper or a lower limit ; a range of values ; an average value ; a median value , a mean value , or a value as compared to a particular control or baseline value . a reference value can be based on an individual sample value such as , for example , a value obtained from a sample from the subject being tested , but at an earlier point in time . the reference value is preferably based on a large number of samples . as used herein , “ fat4 inactivating mutations ” designate any mutations altering the polypeptide sequence of the fat4 protein that significantly reduce its biological activity . these mutations can be non - sense mutation or missense mutations , leading to the generation of truncated fat4 polypeptide to an inactive polypeptide ( e . g ., a mutation in the binding domain to amotl1 ). some inactivating mutations have been disclosed in cappello et al , 2013 and in alders et al . as used herein , “ yap1 or amotl1 inactivating mutations ” are for example any mutations altering their nuclear localisation ( e . g ., mutations in the interacting domain with fat4 ). more precisely , these mutations may prevent their exit from the nucleus or may induce their translocation in the nucleus . preferably , if the yap1 polypeptide contains a mutation that enhances its nuclear localisation , then said mammal is suffering from or will develop cardiac hypertrophy . preferably , if the amotl1 polypeptide contains a mutation that enhances its nuclear localisation , then said mammal is suffering from or will develop cardiac hypertrophy . when , according to the method of the invention , a mammal is diagnosed as suffering from cardiac hypertrophy , any appropriate treatment reducing heart growth or heart size can be provided . traditional treatments that may be used involve e . g ., blocking neurohormones ( catecholamines , angiotensin , aldosterone ), or calcium triggers ( l - type ca 2 + - channel blockers ) or target pathological load ( vasodilators and diuretics ). alternatively , it will be advantageous to treat said mammal by upregulating fat4 or down - regulating yap1 and / or amotl1 , as proposed in the above treating methods of the invention . accordingly , said down - regulation can be carried out by administering an effective amount of a sirna targeting yap1 ( such as those having the sequence seq id no : 24 , seq id no : 25 or seq id no : 27 ) and / or amotl1 ( such as those having the seq id no : 21 to 23 ). alternatively , yap1 down - regulation can be carried out by administering an effective amount of verteporfin or of any chemical compound inhibiting yap1 biological activity . in a preferred embodiment , said mammal is a human . in another preferred embodiment , said human is suspected of suffering from cardiac hypertrophy ( for example , its left ventricle has an abnormal increased size , or an increased thickness or an increased cavity size ). the normal lv mass in men is 135 g and the mass index often is about 71 g / m 2 . in women , the values are 99 g and 62 g / m 2 , respectively . left ventricle hypertrophy is usually suspected when it presents two standard deviations above normal . the typical echo - cardiographic criteria for suspecting left ventricle hypertrophy are thus ≧ 134 and 110 g / m 2 in men and women respectively ( see albergel am . j . cardiol . 1995 , 75 : 498 ). in another preferred embodiment , said mammal is embryonic or newborn . if it is newborn , it is more preferably one month or less of age , one week or less of age , or one day or less of age . in another aspect , the present invention relates to a method , comprising analyzing a tissue sample from a mammal for a fat4 mutation , wherein , if the mutation is present , treating the mammal to prevent or reduce cardiac hypertrophy or heart failure . in a preferred embodiment , said treatment comprises upregulating fat4 , deleting yap , or administering an effective amount of verteporfin . in other aspects , the present invention relates to methods , comprising administering compounds to a fat4 mutant mammal , monitoring cardiac hypertrophy or regeneration in the fat4 mutant mammal , and selecting a compound demonstrating reduction or prevention of cardiac hypertrophy or regeneration or repair in the fat4 mouse mutant or amplification of cardiomyocyte populations . in a preferred embodiment , the fat4 mutant mammal is a fat4 mouse mutant . in another preferred embodiment , said fat4 mutant mammal is embryonic or newborn . if it is newborn , it has more preferably one month or less of age , one week or less of age , or one day or less of age . in a preferred embodiment , said monitoring comprises quantifying cell proliferation and / or cell shape . more precisely , the present invention relates to a screening method for identifying compounds that are useful for preventing and / or treating cardiac hypertrophy , said method comprising the following steps : a ) administering a candidate compound to a transgenic mammal being deficient for the fat4 gene , b ) monitoring cardiac hypertrophy or regeneration in said transgenic mammal before and after step a ), and c ) selecting the candidate compound if it induces the reduction of cardiac hypertrophy in said transgenic mammal . in a preferred embodiment , said step b ) involves the monitoring of the expression of yap1 dependent genes , such as aurkb , ccna2 , birc2 , birc5 , cdkn1b , lyh6 , or acta1 . in this case , it is concluded that the candidate compound leads to the “ reduction of hypertrophy ” when the expression of yap1 dependent genes is reduced in its presence ( as compared with the expression of the same genes prior to its administration ). conversely , it is concluded that the candidate compound leads to “ cardiac growth or regeneration ” when the expression of yap1 dependent genes is enhanced in its presence ( as compared with the expression of the same genes prior to its administration ). in another preferred embodiment , said step b ) comprises quantifying cardiomyocyte proliferation and / or shape . in this case , reduction of hypertrophy is observed when cardiomyocyte proliferation is decreased or when cardiomyocyte size is reduced in the presence of the tested compound . conversely , an enhanced cardiomyocyte proliferation or size will be a sign of cardiac growth or regeneration so that the candidate compound will not be useful for preventing and / or treating cardiac hypertrophy . in a preferred embodiment , the transgenic mammal used in the screening method of the invention is a knock - out fat4 −/− or fat4 flox / flox mammal . preferably , said mammal is any mammal with the exception of human . for example , it is a knock - out fat4 −/− mouse or a knock - out fat4 −/− rat . in a preferred embodiment , said transgenic mammal is embryonic or newborn , and is preferably having one month or less of age , one week or less of age , or one day or less of age . in another embodiment , the screening method of the invention is not carried out on a whole animal but rather on cells extracted therefrom . in this case , the screening method of the invention comprises the following steps : a ) contacting in vitro a candidate compound to at least one cell which is deficient for the fat4 gene , b ) monitoring the proliferation and / or the size of said at least one cell before and after step a ), and c ) selecting the candidate compound if it is able to reduce the proliferation and / or the size of said at least one cell . in a preferred embodiment , said at least one cell is a cardiomyocyte . in another preferred embodiment , said at least one cell is a fat4 −/− or fat4 flox / flox human , mouse or rat cardiomyocyte . the candidate compound is useful for preventing and / or treating cardiac hypertrophy if the proliferation of said at least one cell is decreased or if its size is reduced in its presence ( as compared with in its absence ). conversely , an enhanced proliferation or size will be a sign of cardiac growth or regeneration so that the candidate compound will not be useful for preventing and / or treating cardiac hypertrophy . cell proliferation and / or size may be assessed by any conventional means , such as microscopy analysis , cell counting , labeling of proliferation markers by immunohistochemistry or flow cytometry etc . or monitoring the expression of cell cycle genes . in another embodiment , the screening method of the invention involves the monitoring of the expression level of the modulators of the hippo pathway ( yap1 , amotl1 and / or fat4 ) in cardiomyocyte cells . as a matter of fact , if , in cardiomyocytes contacted with a candidate compound , fat4 expression is enhanced , or if yap1 or amotl1 expression is reduced , then said candidate compound is useful for preventing and / or treating cardiac hypertrophy . one could also look for compounds which will enhance or suppress the proliferative effect of nlsamotl1 ( adenovirus cloned with a nuclear amotl1 ). in a preferred embodiment , the screening method of the invention therefore comprises the following steps : a ) contacting a candidate compound with at least one cardiomyocyte cell , b ) monitoring the expression level of fat4 in said cell before and after step a ), and c ) selecting the candidate compound if contacting said cell with said candidate compound enhances fat4 expression . a ) contacting a candidate compound with at least one cardiomyocyte cell , b ) monitoring the expression level of yap1 and / or amotl1 or the yap1 and / or amotl1 subcellular localisation , in said cell before and after step a ), and c ) selecting the candidate compound if contacting said cell with said candidate compound reduces yap1 and / or amotl1 expression or increase the sequestration of amotl1 or yap1 out of the nucleus . the expression level of fat4 , amotl1 and / or yap1 or the subcellular localisation of these polypeptides may be assessed by any conventional means ( e . g ., by rt - qpcr , elisa , immunohistochemistry , etc .). in another aspect , the present invention relates to a screening method for identifying compounds that are useful for increasing heart size or inducing heart regeneration or for amplifying cardiomyocyte populations , said method comprising the following steps : a ) contacting a candidate compound with at least one cardiomyocyte cell , b ) monitoring the expression level of fat4 in said cell before and after step a ), and c ) selecting the candidate compound if contacting said cell with said candidate compound reduces fat4 expression . a ) contacting a candidate compound with at least one cardiomyocyte cell , b ) monitoring the expression level of yap1 and / or amotl1 in said cell before and after step a ), or monitoring their subcellular localisation , and c ) selecting the candidate compound if contacting said cell with said candidate compound enhances yap1 and / or amotl1 expression or increases their nuclear translocation or reduces their sequestration out of the nucleus . the expression level of fat4 , amotl1 and / or yap1 or the subcellular localisation of these polypeptides may be assessed by any conventional means ( e . g ., by rt - qpcr , elisa , immunohistochemistry , etc .). the subcellular translocation may be assessed by any conventional means ( e . g ., immunohistochemistry , imagestream , etc ). the present invention finally relates to kits comprising the means to detect the expression level of fat4 , yap1 and / or amotl1 in cells or the subcellular localisation of yap1 and / or amotl1 . these means can be primers or probes for the specific detection of the presence or absence of the mrna of these markers . these kits may also contain a heat - resistant polymerase for pcr amplification , one or more solutions for amplification and / or the hybridisation step , and any reagent with which to detect the said markers , preferably in cardiomyocytes . according to an embodiment , these kits may alternatively or additionally contain antibodies that are specific of the fat4 , yap1 and / or amotl1 proteins . in this case , the kits of the invention may also contain any reagent adapted for hybridisation or immunological reaction on a solid carrier . these kits may be used in the screening and / or the diagnosing methods of the invention . more precisely , they may be used for diagnosing cardiac hypertrophy in a mammal , or for identifying compounds that are useful for preventing and / or treating cardiac hypertrophy or for increasing heart size or inducing heart regeneration . fig1 . excessive thickness of fat4 mutant hearts . whole mount views ( a ) and histological sections ( b ) of fat4 +/− and fat4 −/− neonatal ( p0 ) hearts . the arrowhead points to the flattened apex and double arrows highlight ventricular wall and septum thickness . ( c ) quantification of the thickening in fat4 +/+ ( n = 7 ), fat4 +/− ( n = 5 ) and fat4 −/− ( n = 6 ) mutants . scale bar : 500 μm . rv , right ventricle ; lv , left ventricle . in all figures , data are presented as means ± standard deviations , normalised to the level of control hearts when appropriate . statistical significance : * p & lt ; 0 . 05 , ** p & lt ; 0 . 01 , *** p & lt ; 0 . 001 . fig2 . fat4 restricts cell proliferation and hypertrophy . immunodetection ( a ) and quantification ( b ) of the number of mitotic cardiomyocytes positive for phosphorylated histone h3 ( ph3 ) ( arrowheads ) in fat4 +/+ ( n = 6 ), fat4 +/− ( n = 6 ) and fat4 −/− ( n = 5 ) hearts at p0 . ( c ) percentage of cardiomyocytes per total number of cells , analysed by imagestream , in fat4 flox /+ ( controls , n = 7 ) and fat4 flox /− ; mesp1 cre /+ ( mutants , n = 6 ) hearts at p14 . ( d ) relative transcript levels of cell cycle ( aurkb , ccna2 , cdc20 ), cycle exit ( cdkn1b ) or survival ( birc2 / 5 ) genes in fat4 +/+ ( n = 5 ), fat4 +/− ( n = 5 ) or fat4 −/− ( n = 7 ) hearts at p0 . ( e ) increased number of proliferating ki67 - positive cardiomyocytes , as counted by flow cytometry from primary cell cultures treated with fat4 sirna ( n = 20 ) compared to control ( ctrl ) cultures ( n = 20 ). ( f ) immunodetection of cardiomyocyte cross - sectional area , with an antibody to caveolin3 . ( g ) quantification of this in the interventricular septum indicates cell hypertrophy in fat4 −/− ( n = 6 ) compared with fat4 +/+ ( n = 9 ) hearts at p0 . ( h ) expression of positive ( nppb and acta1 ( skeletal actin )) and negative ( myh6 ( α myosin heavy chain )) hypertrophy markers and of a marker of wall stress ( nppa ) quantified by rt - qpcr in fat4 +/+ ( n = 5 ), fat4 +/− ( n = 5 ) or fat4 −/− ( n = 7 ) hearts at p0 . scale bars : 10 μm . fig3 . fat4 modulates hippo signalling . ( a ) relative expression of yap1 target genes in fat4 +/+ ( n = 5 ), fat4 +/− ( n = 5 ) or fat4 −/− ( n = 7 ) hearts at p0 . ( b ) immunodetection of yap1 localisation in the heart of the indicated genotype at p0 . nuclei are encircled with dashed white lines . ( c ) immunodetection of yap1 localisation in primary cultures of cardiomyocytes treated with the indicated sirna . ( d ) quantification of the increased percentage of nuclei with a strong yap1 signal when fat4 is down - regulated ( n = 6 cultures ) relative to the control ( n = 6 ). ( e ) immunodetection of yap1 ( nuclear , arrows ) localisation in primary cultures of cardiomyocytes transfected with the indicated plasmids . ( f ) quantification of its nuclear to cytoplasmic localisation , which is decreased when fat4 is overexpressed ( n = 14 cells ) relative to control gfp ( n = 9 ). ( g ) the number of proliferating ki67 - positive cardiomyocytes , counted by flow cytometry from primary cell cultures , indicates a genetic rescue of fat4 sirna by yap1 sirna ( n = 20 ctrl , 20 fat4 , 8 yap1 and 8 fat4 + yap1 sirna cultures ). ( h ) quantification of the number of mitotic cardiomyocytes positive for ph3 in fat4 flox /+ ( controls , n = 5 ), fat4 flox /− ; mesp1 cre /+ ( mutants , n = 5 ) and fat4 flox /− ; yap1 flox /+ ; mesp1 cre /+ ( rescue , n = 4 ) hearts at p0 . ( i ) percentage of cardiomyocytes , analysed by imagestream , in fat4 flox /+ ( controls , n = 7 ), fat4 flox /− ; mesp1 cre /+ ( mutants , n = 6 ) and fat4 flox /− ; yap1 flox /+ ; mesp1 cre /+ ( rescue , n = 3 ) hearts at p14 . ( j ) western blot showing normal yap1 phosphorylation at the hippo kinase target site . ( k ) corresponding quantification , from the blot shown in supplementary fig3 b , in fat4 +/+ ( n = 4 ), fat4 +/− ( n = 6 ) or fat4 −/− ( n = 4 ) hearts at p0 . ( l ) immunodetection of the localisation of phospho - resistant yap1 ( yap5sa ) in primary cultures of cardiomyocytes treated with the indicated sirna . ( m ) quantification of its nuclear to cytoplasmic localisation , which is increased when fat4 is down - regulated ( n = 6 cultures ) relative to the control ( n = 6 ). ctrl , control ; ns , no significant difference . scale bars : 5 μm in ( b ), 20 μm in ( c ), 10 μm elsewhere . fig4 . amotl1 mediates fat4 signalling . ( a ) immunodetection of cell junctions ( arrowheads ), marked by n - cadherin ( cadh2 ) and plakophilin2 ( pkp2 ), in e18 . 5 hearts . they are disorganised in the absence of fat4 . ( b ) transmission electron micrographs of fat4 +/+ and fat4 −/− hearts at p0 . gap junctions ( arrowhead ) are absent in mutant hearts , whereas the electron - dense material of desmosomes ( arrows ) in the intercalated discs is abnormally spread . ( c ) immunodetection of amotl1 and yap1 ( white arrowheads ) at similar positions near the cell membrane , marked by vinculin ( vcl ), of cardiomyocytes , marked by α - actinin ( actn2 ) in p0 control hearts . asterisks indicate the striation of the sarcomeres . ( d ) amotl1 is relocalised to cardiomyocyte nuclei ( arrowheads ) in fat4 −/− hearts at e18 . 5 . *, non - cardiomyocyte nuclei . ( e ) quantification of nuclear to cytoplasmic localisation of amotl1 in fat4 +/+ ( n = 3 ), fat4 +/− ( n = 3 ) or fat4 −/− ( n = 3 ). ( f ) the number of proliferating ki67 - positive cardiomyocytes , counted by flow cytometry from primary cell cultures , indicates a genetic rescue of fat4 sirna by amotl1 sirna and lower proliferation with amotl1 sirna ( n = 20 ctrl , 20 fat4 , 16 amotl1 and 16 fat4 + amotl1 sirna cultures ). ( g ) immunodetection of yap1 localisation in primary cultures of cardiomyocytes infected with the indicated adenoviruses . the inset shows nuclear co - localisation with ad - ha -( n1s ) 3 - amotl1 . ( h ) increased number of proliferating ki67 - positive cardiomyocytes , as counted by flow cytometry from primary cell cultures infected with nuclear amotl1 ( ad - ha -( n1s ) 3 - amotl1 , n = 8 ), compared to controls ( ad - gfp , n = 8 ). treatment with yap1 sirna ( n = 8 ) is inhibitory . ( i ) immunoprecipitation ( ip ) of amotl1 - ha from cells transfected or not with amotl1 - ha or fat4 - δecd - flag , which is depleted for the extracellular domain ( ecd ). ib , immunoblot with the indicated antibodies . ( j ) co - localisation of amotl1 - ha , fat4 - δecd - flag and endogenous yap1 in primary cultures of transfected cardiac cells . the inset shows an enlargement of the boxed region . scale bars : 0 . 2 μm in ( b ), 5 μm in ( c ), 10 μm elsewhere . ctrl , control . fig5 . model for the role of fat4 in restricting heart growth . fat4 is required to organise cell junctions and sequester amotl1 , preventing excessive heart growth . in the absence of fat4 , amotl1 is released and , in a complex with yap1 , translocates to the nucleus , bypassing the hippo kinases . resulting variations in gene expression promote proliferation and hypertrophy of cardiomyocytes , leading to excessive growth of the myocardium . darker red and yellow indicates high levels of yap1 and amotl1 ; lower levels are indicated by a paler colour . fig6 . fat4 does not impair the coordination of cell divisions in the embryonic heart . ( a ) example of e10 . 5 cardiomyocytes , with labelled nuclei ( blue ), membranes ( red ) and cytoplasmic bridge ( green ). the dotted line indicates the axis of cell division . scale bar : 10 μm . ( b ) distribution of the angle between the axis of cell division and the local transmural axis . in both fat4 +/+ and fat4 −/− hearts , the observed distribution ( blue ), which is significantly different ( wilcoxon u - test two - tailed , p & lt ; 0 . 001 , n = 335 and 524 respectively ) from a random spherical distribution ( red ), indicates a planar bias of the orientation of cell division . ( c ) maps of the regions where the axes of cell divisions were significantly coordinated in the plane of the heart surface . the maps , shown as xy projections , summarize axial data from 4 fat4 +/+ and 3 fat4 −/− hearts at e10 . 5 registered on a single template . isolines delineate regions with an increasing quality of axial coordination ( colour coded ). the blue ellipse shows the average orientation of cell division in the most extensive region , with the corresponding elongation value 1 -( e 2 / e 1 ). the green bars indicate the local average direction per 100 μm × 100 μm box , shifted every 20 μm . nb , number ; lv : left ventricle ; rv : right ventricle . fig7 . proliferation of cardiac cells and rna interference . ( a ) quantification of the number of mitotic non - cardiomyocytes positive for ph3 in fat4 +/+ ( n = 6 ), fat4 +/− ( n = 6 ) and fat4 −/− ( n = 5 ) hearts at p0 ( see fig2 a ). ( b ) immunodetection in fat4 +/+ and fat4 −/− hearts at p0 of cardiomyocytes , marked by cardiac troponin i ( tnni3 ), undergoing cytokinesis , marked by cytoplasmic bridges ( arrowheads ) positive for aurorab kinase ( aurkb ) and acetylated tubulin ( ac - tub ). ( c ) quantification of the number of nuclei per cardiomyocyte in control fat4 flox /+ ( n = 6 ) and mutant fat4 flox /− ; mesp1 cre /+ ( n = 6 ) hearts at p14 . ( d ) histological sections of control fat4 flox /− and mutant fat4 flox /− ; wt1 cre /+ hearts at p0 . ( e ) quantification of the number of mitotic cardiomyocytes positive for ph3 in fat4 flox /+, ( controls , n = 5 ) and fat4 flox /− ; wt1 cre /+ ( mutants , n = 5 ) hearts at p0 . ( f ) efficient down - regulation of fat4 ( n = 6 ctrl , 4 fat4 sirna - 1 , 6 sirna - 2 , 6 sirna - 3 cultures ), yap1 ( n = 6 ctrl , 6 yap1 sirna - 1 , 5 sirna - 2 , 6 sirna - 3 ) or amotl1 ( n = 5 ctrl , 6 amotl sirna - 1 , 6 sirna - 2 , 4 sirna - 3 ) relative to gapdh transcripts , in primary cultures of cardiomyocytes treated with 3 different sirna for each gene . ( g ) western blot showing efficient down - regulation of yap1 and amotl1 proteins relative to gapdh in cultures treated with the indicated sirna . ( h ) corresponding quantifications ( n = 3 cultures in each condition ). ( i ) profile by flow cytometry of primary cell cultures stained with ki67 and tnnt2 ( cardiac troponin t ), using an isotype antibody as a negative control ( green ) or a specific primary antibody ( red ). ( j ) number of proliferating ki67 - positive cardiomyocytes ( tnnt2 - positive ) after treatment with different fat4 , yap1 and amotl1 sirnas ( n = 20 cultures for ctrl and fat4 sirna - 1 , n = 4 in each other condition ). ( k ) immunodetection of replicating edu - positive cardiomyocytes when fat4 is down - regulated , with the corresponding quantification in ( l ) ( n = 6 cultures in each condition ). ctrl , control . scale bar : 5 μm in ( b ), 500 μm in ( d ), 10 μm elsewhere . fig8 . normal canonical hippo signalling when fat4 expression is impaired . ( a ) ( m ) control of yap1 immunodetection in cardiomyocytes treated with the indicated sirna . ( b ) western blot showing normal yap1 phosphorylation at the hippo kinase target site , in hearts at p0 . see quantification in fig3 k . ( c ) the stability of yap1 is decreased in fat4 −/− ( n = 4 ) compared to control ( n = 4 fat4 −/+ , n = 6 fat4 +/− ) hearts , suggesting feedback mechanisms . ( d ) expression of yap1 is not affected in fat4 −/− ( n = 7 ) compared to control ( n = 5 fat4 −/+ , n = 5 fat4 +/− ) hearts at p0 . ( e ) western blot of extracts of primary cultures of cardiomyocytes treated with fat4 sirna . ( f ) normal yap1 phosphorylation and yap1 levels were quantified when fat4 expression is down - regulated ( n = 7 cultures in each condition ). ( g ) western blot , quantified in ( h ), showing no change in the phosphorylation of the hippo kinases lats1 , 2 , between fat4 +/− ( n = 7 ) and fat4 −/− ( n = 7 ) hearts at p0 . ( i ) western blot , quantified in ( j ), showing no change in the phosphorylation of the hippo kinases mst1 , 2 between fat4 +/− ( n = 7 ) and fat4 −/− ( n = 7 ) hearts at p0 . bands in the same lane of western blots were from the same blot , retreated after washing ( see fig . s5 ). ctrl , control ; ns , no significant difference . scale bars : 10 μm . fig9 . fat4 and amotl1 expression and kinetics of the phenotype of fat4 mutant hearts . ( a ) expression of fat4 in wild - type hearts at e10 . 5 ( n = 3 ), e16 . 5 ( n = 4 ) and p0 ( n = 4 ). data are normalised to the level in e10 . 5 hearts . ( b ) histological sections of control and mutant hearts at e14 . 5 , e16 . 5 and e18 . 5 . ( c ) quantification of phosphorylated histone h3 ( ph3 )- positive cells at e10 . 5 ( n = 3 fat4 −/+ , 2 fat4 +/− , 3 fat4 −/− ), e16 . 5 ( n = 2 , 2 , 2 ), e18 . 5 ( n = 3 , 3 , 5 ) and p0 ( n = 6 , 6 , 5 ). ( d ) relative expression of the β - catenin target snai2 in fat4 +/+ ( n = 5 ), fat4 +/− ( n = 5 ) and fat4 −/− ( n = 7 ) hearts at p0 . ( e ) immunodetection of active β - catenin ( ctnnb1 ) in fat4 +/+ and fat4 −/− hearts at p0 . ( f ) quantification of the aspect ratio between the minor and major axes of cardiomyocytes indicates rounder cells in mutant fat4 flox /− ; mesp1 cre /+ ( n = 112 , 792 cells pooled from 6 hearts ) compared to control fat4 flox /+ ( n = 147 , 843 cells pooled from 7 hearts ) samples at p14 . ( g ) relative transcript levels of genes of the angiomotin family in wild - type hearts ( n = 4 ) at e16 . 5 . scale bars : 500 μm . fig1 . nuclear yap1 is not sufficient to promote the proliferation of neonate cardiomyocytes . fig1 . the proliferative effect of nuclear amotl1 is dependent on yap1 . fig1 . yap1 follows amotl1 in cells . colocalisation at the membrane of cultured caridomyocytes . the vesicular localisation of amotl1 titrates yap1 and prevents its nuclear translocation . the nuclear translocation of amotl1 increases nuclear yap1 . fig1 . how to use amotl1 to stimulate cardiomyocyte proliferation ? amotl1 is required for cell proliferation . amotl1 overexpression is not sufficient for its nuclear translocation . fig1 . the pdz binding domain is not involved in the sequestration of amotl1 out of the nucleus . fig1 . phosphorylation by lats2 is not involved in the sequestration of amotl1 out of the nucleus . fig1 . amotl1 is sequestered out of the nucleus via its n - terminal domain . the fat4 mouse mutant line 8 was maintained in a 129s1 genetic background . fat4 conditional mutants 8 were crossed to mesp1 cre /+ 30 , wt1 cre /+ 31 lines or yap conditional mutants 32 and backcrossed in the 129s1 genetic background . fat4 −/− mutants die at birth , whereas fat4 flox /− ; mesp1 cre /+ survive . animal procedures were approved by the ethical committee of the institut pasteur and the french ministry of research . for histological analysis , hearts were excised , incubated in cold 250 mm kcl , fixed in 4 % paraformaldehyde , embedded in paraffin in an automated vacuum tissue processor and sectioned on a microtome ( 10 μm ). for immunofluorescence studies , hearts were fixed in 0 . 5 % paraformaldehyde , embedded in gelatine / sucrose , frozen in cold isopentane and sectioned on a cryostat ( 10 μm ). for the quantification of tissue growth , paraffin sections stained with hematoxylin eosin were imaged on a stereomicroscope . a polygonal mask was drawn in order to isolate the two ventricles from the atria . the green channel ( with highest contrast ) of the resulting image was inverted , thresholded and segmented using connected component analysis . the resulting regions were sorted , retaining the myocardial tissue and excluding blood speckles inside the ventricles , to compute the total area of the ventricles . the penetrance of the myocardial excessive growth was 75 % ( n = 8 ). unless otherwise specified , the image analysis was done using the icy software 33 . cdnas were reverse transcribed from rnas extracted in trizol from cell cultures and isolated hearts using the quanti - tect kit ( qiagen ) and superscript ii reverse transcriptase ( life technologies ) respectively . quantitative pcr was carried out on a stepone system ( life technologies ) using fast start sybr green master ( roche ). quantification of gene expression was calculated as r = 2 δct ( control - target ) , with gapdh used as a control . primers were designed using the ncbi primer - blast software . primer sequences are listed in supplementary table 1 . primary cultures of newborn rat cardiomyocytes , performed as previously described 34 , were transfected with sirna at 10 pm using lipofectamine rnaimax with silencer - sirna at 24 h and analysed at 72 h , or 96 h ( fig3 c ). control ( ambion 4390843 ), fat4 ( sirna - 1 : ambion s172170 , sirna - 2 : ccuguacccugaguauugatt , sirna - 3 ccguccuuguguuuaacgutt ), amotl1 ( sirna - 1 : aucucuaccauuuguugggtt , sirna - 2 : gaguaucucagaggccuautt , sirna - 3 : caucacaugucccagaauatt ), yap1 ( sirna - 1 : ambion s170200 , sirna - 2 : gucagagauacuucuuaaatt , sirna - 3 : ggagaaguuuacuacauaatt ) sirna were used . efficiency of the interference was controlled by rt - qpcr . in the figures , fat4 sirna is fat4 - sirna - 1 , yap1 sirna is a pool of sirna - 1 to 3 and amotl1 sirna is a pool of sirna - 1 to 3 . for flow cytometry analyses , cultures were dissociated to single cell suspensions by trypsin , fixed and permeabilized in ebioscience buffer . proliferating cardiomyocytes were detected by immunostaining with primary antibodies against tnnt2 ( ab64623 ) and ki67 ( bd 556027 ) and counted on a bd lsrfortessa cell analyzer cytometer . gates were set according to isotype control antibodies ( sc - 3887 ). at least 900 cells were counted per condition . alternatively , cardiomyocytes were exposed to edu during 30 h and counted after immunofluorescence ( at least 80 cells per condition ). for overexpression experiments , cardiomyocytes were transfected using lipofectamine 2000 with fat4 - decd - flag ( encoding fat4 depleted for the extracellular domain and for the last c - terminal 297 nucleotides , cb and hmn , unpublished data ), ha - amotl1 24 , yap1 - 5sa ( addgene 27371 ) or control nuclear gfp ( pcig 35 ) plasmids and analysed 24 h later . alternatively , cardiomyocytes were infected with adenoviruses at a multiplicity of infection of 50 and analysed 24 h later , using control ad - gfp 36 or newly generated ha -( nls ) 3 - amotl1 . it was cloned from human amotl1 37 in the adeno - x expression system 3 ( clontech ). immuno fluorescence was performed as previously described 18 , using primary antibodies to acetylated tubulin ( sigma t6793 ), actn2 ( sigma a7811 ), amotl1 ( sigma hpa001196 ), amotl1 ( covalab , gift from d . lallemand ), aurkb ( bd 611082 ), non - phosphorylated ( ser 33 / 37 - thr 41 ) β - catenin ( ctnnb1 , cell signalling 8814 ), cav3 ( bd 610420 ), cdh2 ( ab12221 ), ki67 ( bd 556003 ), mf20 ( dshb ), ph3 ( ab32107 ), pkp2 ( progen 651167 ), scrib ( sc - 28737 ), tnni3 ( ab47003 ), tnnt2 ( ab64623 ), vcl ( sigma f7053 ), yap1 ( sc - 101199 and sc - 15407 ), ha ( roche , 3f10 ), flag ( sigma , f7425 ), alexa fluor conjugated secondary antibodies and hoechst nuclear staining multi - channel 16 - bit images were acquired with a leica sp5 inverted confocal microscope and a 40 / 1 . 25 oil objective or with a zeiss lsm 700 microscope and a 63 ×/ 1 . 4 oil objective . the ph3 channel was thresholded and segmented using connected component analysis , filtering objects under a minimum size of 16 μm 3 in order to eliminate non - specific signals . the myocardial volume of the multi - z scan was estimated by manually outlining the myocardial surface in the median z - slice and computing the area . the total number of cardiomyocyte ( a - actinin - positive ) nuclei in the scan was estimated by manually counting the number of nuclei in a 200 pixels × 200 pixels window extending over all the z - slices , and extrapolating to the total myocardial volume . more than 1 , 500 nuclei were counted per genotype . images of cardiomyocyte transverse sections labelled with caveolin3 ( cav3 ) were acquired systematically in the interventricular septum . cell contours were drawn manually to compute cell area using imagej . at least 40 cells were counted per genotype . the best in - focus z - slice of the hoechst channel was chosen for in vivo cells , whereas in vitro images were z - projected . the analysis involved three image processing steps : 1 ) segmentation of the myocardial ( tnnt2 - positive ) cells using connected component analysis applied after a z - projection ( sum ) and thresholding of the tnnt2 channel ; alternatively , in vitro transfected cells were individually outlined manually ; 2 ) segmentation of the nuclei by thresholding after application of a gaussian filter ( in vivo ), or by the “ active contours ” plugin ( in vitro ); 3 ) measurement of the total intensity of the protein of interest ( pi ) in the tnnt2 - positive cells ( pi tot ) and in their nuclei ( pi nucl ) by multiplication of the pi channel with the respective binary images ( 1 ) and ( 2 ). strong cells were defined as cells in which pi nucl is higher than two standard deviations above the mean pi nucl of control cells . the nuclear / cytoplasmic ratio was computed as : pi nucl / pi cyto = pi nucl /( pi tot − pi nucl ). for in vivo cells , which have a more pronounced 3d shape , total intensities were divided by the area of the segmented object . at least 200 cells were counted per condition . hek293 cells ( q - biogene aes0503 ) were transfected with lipofectamine with the plasmids amotl1 - ha 24 and flag - fat4 - δecd and cultured for 48 h . proteins were extracted in a lysis buffer ( 150 mm nacl , 5 mm edta , 10 mm tris ph 7 . 5 , 10 % glycerol , 1 % np - 40 ) in the presence of protease inhibitors . immunoprecipitation of protein extracts was performed using a standard protocol based on magnetic beads coupled to bacterial protein g , an immunoglobulin - binding protein . proteins were eluted in laemmli buffer . an isotype antibody ( igg ) was used as a negative control of immunoprecipitation . proteins from cell cultures and isolated hearts were extracted for western blots in ripa ( 150 mm nacl , 5 mm edta , 50 mm tris ph 7 . 4 , 0 . 1 % sds , 1 % np - 40 ) and np40 ( 150 mm nacl , 50 mm tris ph 8 , 1 % np - 40 ) buffers , respectively , in the presence of protease and phosphatase inhibitors . equal amounts of proteins were separated on sds - page and transferred to nitrocellulose or pdvf membranes . proteins were detected with the primary antibodies flag ( sigma f1804 ), gapdh ( cell signalling 3683 ), ha ( roche 3f10 ), thr 1079 / 1041 phospho - latsl / 2 ( assay biotech ref a8125 ), lats11 / 2 ( bethyl a300 - 478a ), thr 183 / 180 phospho - mst1 / 2 ( cell signalling 3681 ), mst1 ( cell signalling 3682 ), mst2 ( cell signalling 3952 ), ser 127 phospho - yap1 ( cell signalling 4911 ), yap1 ( cell signalling 4912 ) or amotl1 ( sigma , hpa001196 ), followed by hrp - conjugated secondary antibodies ( jackson immunoresearch ) and the ecl2 detection reagent . protein quantification was obtained by densitometry analysis using a typhoon laser scanner and normalized to gapdh levels . original un - cropped blots are shown in fig1 . in order to obtain a full inferior view of the two ventricles at e10 . 5 , confocal scans of the left ventricle , interventricular region and right ventricle were stitched together . the envelopes of the stitched images were computed by active mesh segmentation 38 . ten such envelopes were used to compute an average envelope ( referred to as the template ), minimising the deformation distances between the template and the envelopes , plus a residual mismatch cost . the metric distance was built on a group of smooth invertible deformations ( i . e . diffeomorphisms 39 ). the axial data from each image were then transported through the deformation between the original envelope and the template , as described by the jacobian matrix of the diffeomorphism ( i . e . the matrix of partial derivatives of the deformation , a 3d generalization of the gradient ). using the polar part of the jacobian was required to avoid improvement of the axial correlation . whole mount immunostaining was carried out on e10 . 5 isolated hearts using scrib and cadh2 antibodies to detect membranes and aurkb antibody to detect cytoplasmic bridges . the nuclei and cytoplasmic bridges were segmented , sister cells were automatically detected and the axes of cell division were calculated as previously described 18 , 40 . for each genotype , at least three e10 . 5 embryonic hearts were registered , in order to pool the axial data on a common template . the planar component of each axis of cell division was extracted by projection over the template envelope . the threshold eigenvalue for each region size , e 1 ( 5 %) , which was obtained by a bootstrap method 18 , was calculated both before and after the diffeomorphic transport of the axes , and the highest value was retained to compensate for any spurious improvement of the alignment due to the transport . contour maps of axial coordination were produced as follows : 1 ) selection of the region , containing at least 50 axes , with the highest eigenvalue e 1 ( core region ); 2 ) listing all regions that both included the core region and had an eigenvalue e 1 & gt ; e 1 ( 5 %) ; 3 ) drawing these regions on the template , with contour values equal to the ratio e 1 / e 1 ( 5 %) . neonate hearts were dissected in cold krebs buffer without calcium , and fixed open with 2 % glutaraldehyde in cacodylate buffer ( na cacodylate 150 mmol / l , cacl2 2 mmol / l , ph 7 . 3 ). the left ventricular papillary muscles were excised and fixed again in 2 % gluteraldehyde in cacodylate buffer , post - fixed in 1 % oso4 , contrasted in 1 % uranyl acetate , dehydrated and embedded into durcupan . ultrathin ( 58 - 60 nm ) longitudinal sections were cut by power - tome mt - xl ( rmc / sorvall , usa ) ultramicrotome , placed on copper slot grids covered with formwar and stained with lead citrate . the sections were examined in a jem 2000fx ( jeol , japan ) electron microscope and recorded using a gatan dualvision 300w ccd camera ( gatan inc ., usa ). p14 hearts were collected , minced and flash frozen as previously described 41 . the defrosted tissue was fixed in 4 % paraformaldehyde , digested with 3 mg / ml collagenase type ii in hbss and filtered using a 100 μm cell - strainer . staining of isolated cells was performed with the bd cytofix / cytoperm fixation / permeabilization kit , using anti - sarcomeric α - actinin ( sigma ) and draq5 nuclear stain . data acquisition was performed using an imagestreamx cytometer with inspire software ( amnis ). files were collected with a cell classifier applied to the brightfield channel to capture events larger than 100 μm . at least 23 , 000 cell events were acquired for each sample and all images were captured with the 40 × objective . data analysis was performed with ideas software ( v6 . 0 , amnis ). images were compensated using a matrix generated by single - stained samples acquired with identical laser settings in the absence of brightfield illumination . the analysis was restricted to in - focus single cells and to intact cardiomyocytes , selected as actinin and draq5 double positive . an object mask was created on the brightfield channel and the aspect ratio was defined as the ratio between the minor and major cell axis . the number of nuclei per cell was assessed using the draq5 images , in at least 350 cells per heart . sample size was chosen in order to ensure a power of at least 0 . 8 , with a type i error threshold of 0 . 05 , in view of the minimum effect size that was looked for . the sample size was calculated using the observed variance of the wild - type mice for the phenotype considered . sample outliers were excluded according to the thompson tau test . the experiments were not randomized and the investigators were not blinded to allocation during experiments and outcome assessment . comparisons of center - values were done on either the average or the geometrical mean when ratios were compared . an anova was systematically calculated when more than two center - values were compared , and tukey - kramer &# 39 ; s test was used for the assessment of bilateral significance . otherwise , a student test was used . when n & gt ; 10 , normality was checked by a kolmogorov - smirnov test or by visualisation of the distribution . when the test was not positive , a wilcoxon u test was used . for quantitative data , when the number of observations n & lt ; 5 , the figures display all data points . for data shown as representative images , the number of replications of the experiments are : 1 experiment ( fig4 b [ n = 1 fat4 +/+, 2 fat4 +/− and 2 fat4 −/− hearts ], 4 g [ 3 cultures ], s 4 e [ n = 2 fat4 +/+ and 3 fat4 −/− hearts ]), 2 experiments ( fig3 b [ n = 3 fat4 +/+ and 3 fat4 −/− hearts ], 4 a [ n = 2 fat4 +/+ and 2 fat4 −/− hearts ], 4 c [ n = 3 fat4 +/+ hearts ], s 2 b [ n = 4 fat4 +/+, 2 fat4 +/− and 2 fat4 −/− hearts ]), 3 experiments ( fig4 i , 4 j ), 4 experiments ( fig8 a ), 50 litters ( fig1 a ), 3 litters ( fig7 d ), 1 litter at e14 . 5 , 3 litters at e16 . 5 , 2 litters at e18 . 5 ( fig9 b ). in the heart , the phenotype of fat4 −/− mutants was assessed at birth . an abnormal flattened apex ( fig1 a ) and 1 . 5 fold thicker ventricular myocardium and interventricular septum ( fig1 b - c ) was observed , compared to control hearts . this excessive tissue thickness may result from defective orientation or excessive rate of growth . as it has been shown previously that clonal growth of the heart is oriented 17 , 18 , the orientation of cell division was therefore analysed in fat4 −/− mutants . whereas in the mouse kidney orientation of cell division is disrupted when fat4 is absent 8 , cell division was found still biased in the plane of the heart surface and still coordinated in the left ventricle of fat4 −/− hearts as in controls ( fig6 ). it was then investigated whether the excessive growth of fat4 mutant hearts was due to increased cell proliferation , and found that the number of cardiomyocytes positive for the mitotic marker , phosphorylated histone h3 , was significantly higher in the absence of fat4 ( fig2 a - b ), whereas the effect was milder in non - cardiomyocytes ( fig7 a ). mutant cardiomyocytes undergo cytokinesis , as revealed by aurkb staining ( fig7 b ). conditional deletion of fat4 , in cardiac progenitors that express mesp1 , resulted in an increased percentage of cardiomyocytes , whereas their number of nuclei was unchanged ( fig2 c , fig7 c ). conditional deletion of fat4 in non - cardiomyocytes , using wt1 - cre , did not change the number of mitotic cells ( fig7 d - e ), indicating that fat4 is required in cardiomyocytes . analysis of gene expression showed an increase for cell cycle ( ccna2 , cdc20 ), cytokinesis ( aurkb ) and anti - apoptotic ( birc2 / 5 ) genes and a decrease of cdkn1b transcripts implicated in cell cycle exit , in fat4 −/− compared to control hearts ( fig2 d ). heterozygotes also show transcript upregulation , although they do not have a detectable heart phenotype , indicating compensation at the level of the proliferation gene network dependent on fat4 dosage . in primary cultures , knock - down of fat4 ( fig7 f - j ) significantly enhanced the number of proliferating ki67 - positive and replicating edu - positive cardiomyocytes ( fig2 e and fig7 k - l ). it was next examined whether fat4 also affects cell size . by measuring the cross sectional area of cardiomyocytes , a significant increase of cell size was found in fat4 −/− compared to control hearts ( fig2 f - g ). in agreement with a hypertrophic phenotype , rt - qpcr revealed a de - regulation of classical markers of heart hypertrophyl 9 , corresponding to the activation , in fat4 −/− mutant hearts , of genes normally expressed at fetal stages ( acta1 ), whereas genes normally expressed at adult stages ( myh6 ) are down - regulated ( fig2 h ). the early marker of heart hypertrophy , nppb 20 , was strikingly increased ( 11 fold ) in fat4 −/− mutant hearts , whereas the marker of wall stress , nppa , was not . these data show that fat4 is required to restrict heart growth at birth , with an effect on the proliferation and hypertrophy of cardiomyocytes . since misexpression of genes ( fig2 d ) previously shown to be targets of the hippo pathway in the control of cardiomyocyte proliferation 1 , 3 was observed , it was examined whether hippo signalling was impaired in fat4 mutant hearts . the classical targets of yap1 , ctgf and cyr61 , were significantly overexpressed ( fig3 a ), showing that the transcriptional activity of hippo effectors is increased in fat4 −/− mutant hearts . consistent with this , yap1 was relocalised to the nucleus of cells in which fat4 was down - regulated , both in vivo ( fig3 b ) and in primary cell cultures ( fig3 c - d , fig8 a ), and reduced in the nucleus of cells in which fat4 was overexpressed ( fig3 e - f ). knock - down of yap1 transcripts by rna interference ( fig7 f - j ) rescued the increased cell proliferation observed when fat4 is down - regulated ( fig3 g ). deletion of one copy of yap1 similarly rescued the excessive number of mitotic cells in mesp1 - cre conditional fat4 mutant hearts , as well as the percentage of cardiomyocytes ( fig3 h - i ). these rescue experiments are consistent with the proposition that fat4 acts upstream of yap1 . to examine whether canonical hippo signalling was activated , as in the fly model , the phosphorylation of yap1 at the ser 127 target site of hippo kinases was analysed . neither in vivo , nor in primary cell cultures ( fig3 j - k , fig8 b - f ), could be detected a significant change in the ratio of phosphorylated yap1 , over total yap1 , when fat4 was down - regulated . the phosphorylation of the hippo kinases lats and mst was also unaffected when fat4 is absent ( fig8 g - j ). a phospho - resistant form of yap1 was also relocalised to the nucleus when fat4 was down - regulated ( fig3 l - m ), further supporting the conclusion that the nuclear translocation of yap1 downstream of fat4 is not mediated by a change of phosphorylation . the phenotype of fat4 mutants differs from that resulting from impairment of the canonical hippo pathway . the onset of excessive myocardial growth in yap1 gain - of - function mutants or in hippo kinase - deficient hearts 1 is already seen at embryonic stages ( e10 . 5 - e11 . 5 ). in contrast , the phenotype of fat4 −/− hearts is detected much later , from e18 . 5 , although fat4 is expressed throughout heart development ( fig8 a - c ). the wnt pathway , which was previously shown to interact with the canonical hippo pathway , was not found activated in fat4 −/− mutants ( fig8 d - e ). these observations show that fat4 is a later modulator of the hippo pathway and suggest that a mechanism other than that of phosphorylation by the hippo kinases , regulates the nuclear localisation of yap1 , downstream of fat4 . hippo signalling is modulated by cell junction proteins 21 . when cardiomyocytes were labelled with junction markers , abnormal cell junctions were observed in fat4 −/− hearts . n - cadherin ( cadh2 ) or plakophilin2 ( pkp2 ) staining were broader and less focalised than in control hearts ( fig4 a ). by electron microscopy , the electron dense desmosomal material was more diffuse and no gap junctions were detected in cardiomyocytes of fat4 −/− hearts ( fig4 b ). in agreement with abnormal cell junctions , cardiomyocytes had a rounder shape in mesp1 - cre conditional fat4 mutant hearts ( fig8 f ). these observations suggest that a junctional protein may be involved in the effect of the atypical cadherin fat4 on hippo effectors . it was focussed on the adaptor protein , angiomotin , which had been shown to interact with yap1 and to co - translocate to the nucleus , where the complex modulates transcription 22 . this co - translocation can occur independently of yap1 phosphorylation 23 . in the heart , it was observed that angiomotin - like 1 ( amotl1 ), another member of the angiomotin family , is predominantly expressed ( fig8 g ). it is present near the membrane of cardiomyocytes , in clusters where yap1 is also detected , between the z - lines of the sarcomere ( fig4 c ). strikingly , amotl1 was relocalised to the nucleus when fat4 was absent ( fig4 d - e ). when it was interfered with amotl1 ( fig7 f - j ) as well as fat4 expression , the increased cell proliferation observed when fat4 expression alone is down - regulated was reversed ( fig4 f ). interference with amotl1 expression alone shows that it is required for the proliferation of cardiomyocytes . on the contrary , forcing amotl1 to translocate to the nucleus , by addition of a nuclear localisation signal , resulted in co - accumulation of yap1 in the nucleus and stimulation of cardiomyocyte proliferation ( fig4 g - h ). interference with yap1 expression shows that the proliferative effect of nuclear amotl1 is dependent on yap1 . both fat4 and amotl1 are known to interact with the scaffold multi - pdz domain protein , mpdz , also known as mupp1 24 , 25 . it is now shown , in transfected hek293 cells , that amotl1 interacts physically with fat4 , as well as yap1 ( fig4 i ), and that the three proteins co - localise in transfected cardiac cells ( fig4 j ). this is consistent with amotl1 release from cell junctions when fat4 is absent . these results identify amotl1 as a modulator of the hippo pathway in cardiomyocytes , and show that it is prevented from entering the nucleus by sequestration in a fat4 complex , thus restricting yap1 mediated tissue growth . these observations of yap1 activity in fat4 mutants , as well as the suppression of fat4 dependent hyperproliferation by reduced yap1 or amotl1 expression , argue that fat4 is an upstream regulator of yap1 in the mouse heart , and that it triggers a non - canonical modulation of hippo signalling . this pathway probably implicates a non - phosphorylated form of yap1 bound to amotl1 in the cytoplasm . when amotl1 is not sequestered at cell junctions with fat4 , it was shown that it is an intermediate , that bypasses the hippo kinases , to regulate the nuclear translocation of yap1 . amotl1 may also directly contribute to the transcriptional activation of target genes , by analogy with amot in the liver 22 . it remains to be seen whether tead , a transcription factor that interacts with yap1 , is implicated in this context . this model is shown in fig5 . amotl1 has no homologue in flies , which explains why the intracellular domain of fat4 cannot rescue the growth phenotype of fat mutant flies 12 . the function of mouse amotl1 is similar to that of drosophila expanded , a ferm - domain protein which requires fat for its localisation at the membrane 5 and which can directly sequester yorkie out of the nucleus , independently of canonical hippo signalling 26 . the mammalian homologue of expanded , frmd6 , has lost the c - terminal domain of interaction with hippo effectors , which supports an evolutionary switch in the regulation of hippo signalling by fat 16 . although fat signalling is implemented differentially between mouse and fly , the function of this cadherin is well conserved , with a dual effect on tissue polarity 8 and also , as the present inventors show , on tissue growth . the effect of fat4 depends on the cellular context . in the heart , it was shown that fat4 regulates tissue growth , rather than polarity . this has also been observed in the cortex 11 , whereas in other organs , such as the kidney or the cochlea , fat4 is a regulator of tissue polarity 8 , 9 . these findings on fat4 mutants uncover a mechanism that restricts heart growth at birth . central to this mechanism is the adaptor protein amotl1 , which can shuttle from cell junctions to the nucleus , transporting the transcription co - factor yap1 . whereas the hippo pathway was shown to be required at embryonic stages of heart development 1 , 2 , fat4 is a later modulator exerting its role at birth . it remains to be established how the fat4 / amotl1 dependent pathway is activated and what is its relative importance to regulate yap1 , in comparison with canonical hippo signalling . canonical hippo signalling is also modulated by cell junctions in cardiomyocytes , where remodeling of the intercalated discs activates hippo signalling , with pathological consequences leading to arrhythmogenic cardiomyopathy 27 . fat4 mutants display hypertrophy , in addition to increased cell proliferation . although hypertrophy can potentially be induced by yap1 4 , 28 , other studies 2 , 3 would suggest that this is an indirect effect . due to its positive effect on cardiomyocyte proliferation , hippo signalling has been shown to be important for prolonging the regenerative potential of the mouse heart 14 , 15 , which normally ceases during the first week after birth 29 . however , yap1 is less efficient in promoting cardiomyocyte proliferation at postnatal stages than it is during development , which suggests that other factors block yap1 activity at later stages . it was now identified upstream regulators of yap1 activity in the heart and it can be anticipated that manipulating the fat4 pathway will facilitate the reactivation of cardiomyocyte proliferation induced by phospho - resistant yap1 15 or hippo kinase deficiency 14 . this has major therapeutic implications for the repair of the failing human heart . 1 . heallen , t . et al . hippo pathway inhibits wnt signaling to restrain cardiomyocyte proliferation and heart size . science 332 , 458 - 461 ( 2011 ). 2 . xin , m . et al . regulation of insulin - like growth factor signaling by yap governs cardiomyocyte proliferation and embryonic heart size . sci signal 4 , ra70 ( 2011 ). 3 . von gise , a . et al . yap1 , the nuclear target of hippo signaling , stimulates heart growth through cardiomyocyte proliferation but not hypertrophy . proc natl acad sci usa 109 , 2394 - 2399 ( 2012 ). 4 . del re , d . p . et al . yes - associated protein isoform 1 ( yap1 ) promotes cardiomyocyte survival and growth to protect against myocardial ischemic injury . j biol chem 288 , 3977 - 3988 ( 2013 ). 5 . silva , e ., tsatskis , y ., gardano , l ., tapon , n . & amp ; mcneill , h . the tumor - suppressor gene fat controls tissue growth upstream of expanded in the hippo signaling pathway . curr biol 16 , 2081 - 2089 ( 2006 ). 6 . fanto , m . et al . the tumor - suppressor and cell adhesion molecule fat controls planar polarity via physical interactions with atrophin , a transcriptional co - repressor . development 130 , 763 - 774 ( 2003 ). 7 . ma , d ., yang , c . h ., mcneill , h ., simon , m . a . & amp ; axelrod , j . d . fidelity in planar cell polarity signalling . nature 421 , 543 - 547 ( 2003 ). 8 . saburi , s . et al . loss of fat4 disrupts pcp signaling and oriented cell division and leads to cystic kidney disease . nat genet 40 , 1010 - 1015 ( 2008 ). 9 . mao , y . et al . characterization of a dchs1 mutant mouse reveals requirements for dchs1 - fat4 signaling during mammalian development . development 138 , 947 - 957 ( 2011 ). 10 . bossuyt , w . et al . an evolutionary shift in the regulation of the hippo pathway between mice and flies . oncogene ( 2013 ). 11 . cappello , s . et al . mutations in genes encoding the cadherin receptor - ligand pair dchs1 and fat4 disrupt cerebral cortical development . nat genet 45 , 1300 - 1308 ( 2013 ). 12 . pan , g . et al . signal transduction by the fat cytoplasmic domain . development 140 , 831 - 842 ( 2013 ). 13 . li , f ., wang , x ., capasso , j . m . & amp ; gerdes , a . m . rapid transition of cardiac myocytes from hyperplasia to hypertrophy during postnatal development . j mol cell cardiol 28 , 1737 - 1746 ( 1996 ). 14 . heallen , t . et al . hippo signaling impedes adult heart regeneration . development 140 , 4683 - 4690 ( 2013 ). 15 . xin , m . et al . hippo pathway effector yap promotes cardiac regeneration . proc natl acad sci usa 110 , 13839 - 13844 ( 2013 ). 16 . huang , j ., wu , s ., barrera , j ., matthews , k . & amp ; pan , d . the hippo signaling pathway coordinately regulates cell proliferation and apoptosis by inactivating yorkie , the drosophila homolog of yap . cell 122 , 421 - 434 ( 2005 ). 17 . meilhac , s . m ., esner , m ., kerszberg , m ., moss , j . e . & amp ; buckingham , m . e . oriented clonal cell growth in the developing mouse myocardium underlies cardiac morphogenesis . j cell biol 164 , 97 - 109 ( 2004 ). 18 . le garrec , j . f . et al . quantitative analysis of polarity in 3d reveals local cell coordination in the embryonic mouse heart . development 140 , 395 - 404 ( 2013 ). 19 . izumo , s ., nadal - ginard , b . & amp ; mandavi , v . protooncogene induction and reprogramming of cardiac gene expression produced by pressure overload . proc natl acad sci usa 85 , 339 - 343 ( 1988 ). 20 . nakagawa , o . et al . rapid transcriptional activation and early mrna turnover of brain natriuretic peptide in cardiocyte hypertrophy . evidence for brain natriuretic peptide as an “ emergency ” cardiac hormone against ventricular overload . j clin invest 96 , 1280 - 1287 ( 1995 ). 21 . enderle , l . & amp ; mcneill , h . hippo gains weight : added insights and complexity to pathway control . sci signal 6 , rel ( 2013 ). 22 . yi , c . et al . the p130 isoform of angiomotin is required for yap - mediated hepatic epithelial cell proliferation and tumorigenesis . sci signal 6 , ra77 ( 2013 ). 23 . wang , w ., huang , j . & amp ; chen , j . angiomotin - like proteins associate with and negatively regulate yap1 . j biol chem 286 , 4364 - 4370 ( 2011 ). 24 . sugihara - mizuno , y . et al . molecular characterization of angiomotin / jeap family proteins : interaction with mupp1 / patj and their endogenous properties . genes cells 12 , 473 - 486 ( 2007 ). 25 . ishiuchi , t ., misaki , k ., yonemura , s ., takeichi , m . & amp ; tanoue , t . mammalian fat and dachsous cadherins regulate apical membrane organization in the embryonic cerebral cortex . j cell biol 185 , 959 - 967 ( 2009 ). 26 . badouel , c . et al . the ferm - domain protein expanded regulates hippo pathway activity via direct interactions with the transcriptional activator yorkie . dev cell 16 , 411 - 420 ( 2009 ). 27 . chen , s . n . et al . the hippo pathway is activated and is a causal mechanism for adipogenesis in arrhythmogenic cardiomyopathy . circ res 114 , 454 - 468 ( 2014 ). 28 . wang , p . et al . the alteration of hippo / yap signaling in the development of hypertrophic cardiomyopathy . basic res cardiol 109 , 435 ( 2014 ). 29 . porrello , e . r . et al . transient regenerative potential of the neonatal mouse heart . science 331 , 1078 - 1080 ( 2011 ). 30 . saga , y . et al . mesp1 is expressed in the heart precursor cells and required for the formation of a single heart tube . development 126 , 3437 - 3447 ( 1999 ). 31 . wessels , a . et al . epicardially derived fibroblasts preferentially contribute to the parietal leaflets of the atrioventricular valves in the murine heart . dev biol 366 , 111 - 124 ( 2012 ). 32 . reginensi , a . et al . yap - and cdc42 - dependent nephrogenesis and morphogenesis during mouse kidney development . plos genet 9 , e1003380 ( 2013 ). 33 . de chaumont , f . et al . icy : an open bioimage informatics platform for extended reproducible research . nature methods 9 , 690 - 696 ( 2012 ). 34 . diguet , n . et al . muscle creatine kinase deficiency triggers both actin depolymerization and desmin disorganization by advanced glycation end products in dilated cardiomyopathy . j biol chem 286 , 35007 - 35019 ( 2011 ). 35 . megason , s . g . & amp ; mcmahon , a . p . a mitogen gradient of dorsal midline wnts organizes growth in the cns . development 129 , 2087 - 2098 ( 2002 ). 36 . relaix , f . et al . pax3 and pax7 have distinct and overlapping functions in adult muscle progenitor cells . j cell biol 172 , 91 - 102 ( 2006 ). 37 . pei , z ., bai , y . & amp ; schmitt , a . p . piv5 m protein interaction with host protein angiomotin - like 1 . virology 397 , 155 - 166 ( 2010 ). 38 . dufour , a ., thibeaux , r ., labruyere , e ., guillen , n . & amp ; olivo - marin , j . c . 3 - d active meshes : fast discrete deformable models for cell tracking in 3 - d time - lapse microscopy . ieee transactions on image processing 20 , 1925 - 1937 ( 2011 ). 39 . charon , n . & amp ; trouve , a . the varifold representation of non - oriented shapes for diffeomorphic registration . siam journal on imaging sciences 6 , 2547 - 2580 ( 2013 ). 40 . pop , s . et al . extracting 3d cell parameters from dense tissue environments : application to the development of the mouse heart . bioinformatics 29 , 772 - 779 ( 2013 ). 41 . mollova , m . et al . cardiomyocyte proliferation contributes to heart growth in young humans . proc natl acad sci usa 110 , 1446 - 1451 ( 2013 ). 42 . moak j p1 , kaski j p . hypertrophic cardiomyopathy in children . heart . 2012 july ; 98 ( 14 ): 1044 - 54 . 43 . tariq m1 , ware s m1 . importance of genetic evaluation and testing in pediatric cardiomyopathy . world j cardiol . 2014 nov . 26 ; 6 ( 11 ): 1156 - 65 44 . sudol m1 , shields d c , farooq a . structures of yap protein domains reveal promising targets for development of new cancer drugs . semin cell dev biol . 2012 september ; 23 ( 7 ): 827 - 33 .

the present invention provides methods of treating and preventing cardiac hypertrophy and heart failure . further provided are transgenic animals exhibiting altered expression of the atypical cadherin fat4 and methods using said transgenic animals , or cells isolated therefrom , for the detection of compounds having therapeutic activity toward cardiac hypertrophy or regeneration . embodiments of the present invention provide methods and composition for therapeutic intervention in cardiac hypertrophy or heart repair by modulating fat4 and / or amotl1 . treatment may include deleting yap or administering verteporfm . embodiments of the present invention define the molecular events linking fat4 and amotl1 to cardiac growth , and show that fat4 is required to restrict cardiomyocyte hypertrophy and cardiomyocyte proliferation and that this is mediated by amotl1 .

with reference to fig1 , there is disclosed a pinball machine 10 according to one embodiment of the present invention . the pinball machine includes a box - like frame 12 with a bottom panel 14 and a plurality of side walls . the side walls include a first frame member 16 and a second opposing frame member 18 . the first frame member 16 and second frame member 18 are positioned on opposite ends of the bottom panel 14 . each of the frame members 16 , 18 extend upwardly from the bottom panel 14 so as to define a boundary of the box - like frame 12 . the side walls also include a third frame member 20 and a fourth frame member 22 , which is positioned on the opposite side of the bottom panel 14 as compared to the third frame member . similar to the first frame member and second frame member 16 , 18 , the third frame member and fourth frame member 20 , 22 extend upwardly from the bottom panel 14 and help to define a playing boundary for the pinball machine as will be discussed below . the first frame member 16 is attached at both of its ends to respective ends of the third and fourth frame members 20 , 22 such that the first frame member is substantially perpendicular to the third and fourth frame member . similarly , the second frame member 18 is positioned adjacent to opposing ends of the third frame member 20 and fourth frame member 22 and is also substantially perpendicular to these frame members . as shown in the figure , a curved wall 23 may be positioned between adjacent end walls of each of the frame members such that a smooth transition is achieved between the substantially perpendicular frame members . the bottom panel 14 preferably includes a playing surface 24 and oppositely - facing rear surface 26 . the playing surface 24 sets the stage of how the pinball game is played . a further discussion regarding this feature will follow . the rear surface 26 faces downwardly . extending downwardly from the rear surface 26 of the bottom panel 14 is a plurality of coupling elements 28 . the coupling elements 28 may be formed in conjunction with the rear surface 26 such as during a molding , extruding or shaping process . in an alternate embodiment , the coupling elements 28 may be attached to the rear surface 26 of the pinball machine 10 using various attaching mechanisms such as rivets , screws and the like . in either case , each coupling element 28 of which there are preferably four , includes an exterior wall 30 and an interior wall 32 . the interior wall 32 defines a cavity 34 that is preferably cylindrical . the exterior wall 30 and interior wall 32 are attached to one another by a circumferential edge 33 . although the coupling elements 28 are shown as protrusions extending outwardly from the rear surface 26 , in alternate embodiments , the coupling elements 28 may be recesses disposed within the rear surface 26 . the pinball machine 10 further includes a first set of legs 40 , each having a first end 42 and a second end 44 . each of the legs in the first set of legs 40 is preferably cylindrical and has a diameter , at least at their respective first end 42 that is substantially equal to the diameter of the cavity 34 of the coupling elements 28 . in a method of assembly , the first legs 40 may be received within coupling elements 28 a and 28 b , which are proximate the first frame member 16 of the pinball machine 10 and remote from the second frame member 18 . the first ends 42 of the legs 40 may be received within the cavity 34 . preferably a securing mechanism such as a snap fit or compression fit as well as other methods known to those in the art interlocks the legs 40 within the cavities 34 . for instance , each of the cavities 34 may have an internal thread that can threadly engage external threads disposed on the first end 42 of the legs thereby allowing the legs to be screwed into the coupling elements 28 such that they are secured therewith . the pinball machine 10 further includes a second set of legs 50 , each of which also include first ends 52 and second ends 54 , shown in fig2 . the second set of legs 50 are received within the cavities 34 of coupling elements 28 c ( not shown in fig1 ) and 28d respectively and similar to the first set of legs 40 , the second set of legs 50 may include various mechanisms and / or features that enable the first ends 52 of the second set of legs to be tightly secured within a cavity 34 of the coupling elements 28 c and 28 d . an example of one of the second set of legs 50 is illustrated in fig2 . as shown in fig2 , the leg 51 which is one of the second set of legs 50 includes a first extension 60 and a second extension 62 . the first extension 60 is received within an opening 64 of the second extension 62 such that the two extensions may be telescopically connected . the first extension 60 includes a protrusion 66 extending outwardly from an exterior surface 68 of the first extension . the protrusion may include the ability to be recessed within the exterior surface 68 of the first extension 60 by including various springs and features that enable such a structure . in contrast , the exterior surface 70 of the second protrusion 62 includes a plurality of apertures 72 extending along the exterior surface 70 of the second extension 62 . the plurality of apertures 72 are preferably aligned along a common longitudinal axis , although this is not required . as illustrated in the figure , in one arrangement , the protrusion 66 of the first extension 60 extends outwardly through aperture 72 a of the second extension thereby locking first extension 60 to the second extension 62 . this enables leg 50 a to have a first height h that is equal to height of the first set of legs 40 . in order to extend the length of the second set of legs 50 , the protrusion may be moved inward towards the exterior surface 68 of the leg 50 a thereby allowing the first extension 60 to be moved upward in a direction illustrated by arrow a . the protrusion may then be allowed to extend through aperture 72 b . this increases the overall height of the leg 50 a to a height h , which is greater than the height h . a further discussion regarding this feature is described below . once the pinball machine 10 is assembled with the various legs attached to the coupling elements , the playing surface 24 is substantially planar and horizontal so long as the sets of legs 40 and 50 have a common height . various features of the game will now be described with the assumption that all the legs are at a first height , which is common throughout . in this example , the sets of legs 40 , 50 all extend the same length . in this orientation , the game is specifically adapted to be played by two opposing players . the term “ opposing ” players refers to the ability of the players to score against each other as opposed to each player trying to achieve his own high score and then comparing that score to the other opposing player . the playing surface 24 includes a plurality of ball - engaging mechanisms 70 mounted to the surface . the ball - engaging mechanisms 70 may include a variety of elements such as posts , bumpers and sling - shot bumpers , as well as other similar ball - engaging mechanisms that are well known in the art . the ball - engaging mechanisms 70 may be constructed as discussed in u . s . pat . nos . 5 , 730 , 441 ; 5 , 707 , 059 ; 6 , 000 , 697 and 6 , 113 , 097 , the disclosures of which are hereby incorporated by reference herein . in either case , the ball - engaging mechanisms 70 have a point value assigned to them such when they are struck with a ball 72 during the play of the game , a score is accrued . additional ball - engaging mechanisms 70 of the game may include an element that propels the ball away from the specific ball - engaging mechanisms when contacted by the ball , such as leaf or trigger switches well known in the art . the pinball machine 10 further includes a first plunger 74 , which is biased with a spring ( not shown ) and used to propel a ball 70 onto the playing surface 24 for play . as for instance , a first player may stand at the end of the machine adjacent the first frame member 16 where the first plunger 74 is located . the first player may draw back on the plunger 74 and then let go . the plunger 74 contacts the ball 72 and exerts a force outwardly thereby propelling the ball through runway 76 until the ball 72 is projected outwardly through gate 78 disposed at the end of the runway 76 . the gate 78 is biased such that once the ball propels outwardly from the runway 76 , the gate swings back thereby closing one end of the runway . once the ball 72 exits the gate 78 , it may contact the ball - engaging mechanisms 70 thereby accruing a point value each time it contacts one of the mechanisms . this point value is constantly updated , tallied and reflected at display 80 . display 80 may be an lcd scoring system or various other counters known to those in the art . a general outline of a scoring system known to those in the art is shown in fig4 . for instance , ball - engaging mechanism 70 is positioned on playing surface 24 . when ball 72 comes into contact with ball - engaging mechanism 70 , a signal is sent by the ball - engaging mechanism to a processing center 75 . the signal may be sent via a pathway 77 which may include mechanical actuation members , electrical elements and combinations of the same . as the signals are sent by the ball - engaging mechanism 70 , the processing center 75 computes the individual points awarded and also keeps track of a tallied total . various software known to those in the art may be included within the processing center 75 so as to keep track of the points as the scores continue to tally . the processing center may then send a signal to display 80 such that the players are aware of the total number of points being accrued . although a first player starts of the game , propelling the ball 72 through runway 76 , a second player is positioned adjacent to second frame member 18 . as the points are being tallied , neither the first player nor second player has yet accrued any of these points . during the game , the first player has the ability to control a first set of flippers 82 by compressing actuation buttons 83 disposed on the side walls of the box - like structure 14 . actuation of buttons 83 causes flippers 82 to pivot about an axis such that the front end of the flippers propel forward . button 83 a controls flipper 82 a while button 83 b controls flipper 82 b . the flippers 82 a and 82 b are separated by a distance thereby enabling a ball to pass between the two flippers and into a goal area 84 . similarly , the second player has control over a second set of flippers 86 by the use of actuation buttons 87 a and 87 b . as before , button 87 a controls movement of flipper 86 a while button 87 b controls movement of flipper 86 b . the two flippers are separated by a distance at their respective front ends such that a ball may fit between the two flippers and into a second goal area 88 . therefore , in a two - player orientation , the ball 72 may come in come in contact with ball - engaging mechanisms 70 including various bumpers 70 a and a spinning trap 70 b as well as additional ball - engaging mechanisms known to those in the art . each time the ball engages one of the ball - engaging mechanisms 70 , a point or a plurality of points may be tallied . in order to be awarded the tallied points , a player must score a goal on the opposite player by getting the ball 72 between the players opposing flippers 72 or 82 and into the goal area 84 or 88 . for instance , if the ball should happen to go into goal 88 adjacent to the second player , all the points that were previously tallied during the sequence would go to the first player . a second ball may now be introduced into the playing surface and begin the process again . as such , a single ball being introduced onto the playing surface 24 and exited the playing surface by exiting through one of the goal areas 84 or 88 constitutes a single round . once one player wins the points tallied , the points on the display 80 are awarded to the player and displayed at a different location on display 80 such as first player display 81 on display 80 in fig4 . the second player display 83 illustrates the score of the second player and tally display 85 illustrates the score tallied before it is awarded to one of the players . in order to create a faster game meaning that the ball travels quicker , an additional set of flippers 90 a and 90 b may also be controlled by the buttons 83 a and 83 b by the first player . this additional set of flippers is closer to the center of the playing surface 24 and thereby allows the first player to have more input on the ball projection throughout the playing surface 24 . similarly , the second player may have an additional set of flippers 92 a and 92 b , which are also controlled by the buttons 87 a and 87 b by the second player . it is important to remember that the two - player game is played with all of the legs being equal in height such that the playing surface is substantially horizontal . in an alternate embodiment , the second set of the legs 50 may be extended such that they are at a height h thereby creating a slanted playing surface 124 such that a ball 172 is urged toward the first player by gravity , for instance , as shown in fig5 , a second set of legs 150 has a height h which is greater than the height of the first set of legs 140 thereby giving the box - like construction 112 a slanted axis angled toward a first frame member 116 . like features are discussed using like character references as compared to the first embodiment of the present invention unless specifically articulated . in this orientation , a single player may play the pinball machine 110 similar to a conventional pinball game . in order to aid in this one - player game , as shown in fig6 , a plurality of stopping gates 190 may be positioned within the various open spaces that are adjacent the second frame member 118 , where the second player would stand in a two - player game . the stopping gates 190 prohibit the ball 170 from exiting into a goal on the second player side as well as other runways and exit avenues . the stopping gates 190 may be recessed within the bottom surface 114 of the pinball machine 112 and actuated upwards using various levers and the like . in such an orientation , a single player may switch the score system to a one - player mode such that all the points accrued by the ball 172 contacting the ball - engaging mechanisms 170 are immediately accrued to the single player and continue to tally as the ball engages various different mechanisms . the second set of legs 150 may be extended at different lengths thereby increasing the angle of the playing surface 124 . as the angle increases , the pinball game becomes faster as gravity has more influence on the ball 172 , pulling the ball towards the flippers of the first player . in an alternate embodiment , as shown in fig7 , the pinball machine 210 may have a first set of legs 240 and a second set of 250 that are substantially identical , and have a common height . therefore , in order to slant the box - like frame 112 , a pivoting element 192 may be provided . the pivoting element is attached to a portion of the box - like frame 212 and may pivot along a joint 293 . the pinball machine 210 also includes a support surface 294 disposed proximate first ends of the various legs . the support surface 294 includes a plurality of positioning elements 295 , which may be engaged by the pivoting element 292 . the pivoting element 292 is positioned proximate one of the positioning elements 295 a , as shown in the figure , such that the box - like frame 212 may be tilted at a specific angle . pivoting element 292 may be engaged with additional positioning elements 295 a or 295 c resulting in a different angle at which the box - like frame 212 is positioned . although the invention herein has been described with reference to particular embodiments , it is to be understood that these embodiments are merely illustrative of the principles and applications of the present invention . it is therefore to be understood that numerous modifications may be made to the illustrative embodiments and that other arrangements may be devised without departing from the spirit and scope of the present invention as defined by the appended claims .

a pinball adapted for play by either one or two players . the pinball game includes the playing surface having first and second ends . the playing surface is adapted for supporting a pinball between the ends . the pinball game also includes a tilting assembly associated with the playing surface for orientating the playing surface between a substantially horizontal position and an inclined position directed towards one of the first and second ends . when the pinball game is adapted to be played by one player , the playing surface is in the inclined position . but when the pinball game is adapted to be played by two players , the playing surface is in the horizontal position .

with reference to fig1 , the present invention provides a method for manufacturing biomedical filler materials rich - contained with at least one thrombin factor and at least one growth factor . the method comprises the steps of : providing a reaction container ( step 100 ); providing a plurality of filler materials , and executing a classification and blending toward the filler materials ( step 101 ); covering the filler materials with a dry first powder composed of the thrombin factor , and depositing the filler materials into the reaction container ( step 102 ); and mixing a mixed - liquid with a second powder rich - contained with the growth factor to obtain a mixture , and further homogeneously mixing the mixture with the filler materials covered with the dry first powder , so as to obtain the biomedical filler materials rich - contained with the thrombin factor and the growth factor ( step 103 ). the reaction container is made of acrylic material . the filler materials can be bone - grafts , such as calcium sulfate salts , calcium phosphates salts , etc ., which are capable of being absorbed by human &# 39 ; s body . the filler materials can be provided in one of a tablet condition , a powder condition and a colloid condition . when the filler materials are provided in the powder condition , the filler materials are composed of powder particles size mainly within a distribution range of 50 to 320 meshes . in step 102 , the dry first powder composed of the thrombin factor covers the surface of the filler materials via a physical electronic - adsorption action . the mixed - liquid is selected in one of platelet - rich plasma ( prp ) made from a blood after suffering an ultra - centrifugal condition , normal saline , medical - injection water and medical - grade dilute phosphoric acid . the second powder rich - contained with the growth factor contains 10 % wt ( percentage in weight ) to 90 % wt of thrombin . a mixing proportion of the mixed liquid and the second powder is between 1 : 0 . 1 and 1 : 0 . 5 , wherein the most optimized value of the mixing proportion is 2 : 1 , i . e ., the mixing proportion of 1 : 0 . 5 is the most optimized . the first preferred embodiment of the present invention comprises the steps as follows : ( 1 ) executing classification , blending toward calcium sulfate hemihydrate powder provided by j . t . baker , following astm standard of u . s . a ., so as to classification , blending the calcium sulfate hemihydrate powder composed of powder particles size range mainly of 200 meshes ; ( 2 ) operating a vacuum valve to provide a working pressure of 10 − 3 to 10 − 2 torr , and providing a working voltage of 5 kev , so as to make the dry first powder composed of the thrombin factor adsorb to calcium sulfate hemihydrate powder via the physical electro - adsorption action ; ( 3 ) providing the reaction container ( made of acrylic material ) having at least one recessed space with a length of 5 cm , a width of 2 cm and a depth of 2 cm ; ( 4 ) providing medical - grade injection water and a stirrer , mixing the medical - grade injection water respectively with second powder ( rich - contained with the growth factor ) and pure calcium sulfate hemihydrate powder in the mixing proportion of 2 : 1 to get a first mixture and a second mixture , putting the first mixture and the second mixture into the recessed space of the reaction container , and stirring the stirrer in a rate of 30 revolution per minute ( rpm ) for 15 seconds to make the first mixture and second mixture within the recessed space be sufficiently mixed ; and ( 5 ) preparing the vicat needle , with a tip area of 1 mm 2 and a weight of 300 g , served as test equipment , laying the tip of the vicat needle on the first mixture and second mixture in each interval of 30 seconds until no recessed mark appears on the surface of the first mixture and second mixture , and respectively recording the how many intervals spent until no recessed mark appears on the surface of the first mixture and second mixture , so as to respectively get a first hardening time and a second hardening time . please refer to table 1 , in initial , both the first mixture and the second mixture are in paste status after stirring for 15 seconds ; however , the hardening degree of the first mixture is much different from that of the second mixture . it is obviously that second powder rich - contained with the growth factor has greater hardness . please further refer to fig2 and fig3 , which illustrate two x - ray experiments respectively toward pure calcium sulfate hemihydrate and the second powder rich - contained with the growth factor . it is obvious that there is only one simplified peak for pure calcium sulfate hemihydrate ; while , there are many different peaks for the second powder rich - contained with the growth factor as shown in fig3 . the second preferred embodiment of the present invention comprises the steps as follows : ( 1 ) executing classification , blending toward calcium phosphate powder provided by j . t . baker , following astm standard of u . s . a ., so as to classification blending the calcium phosphate powder composed of powder particles size mainly of 200 meshes ; ( 2 ) operating a vacuum valve to provide a working pressure of 10 - 3 to 10 - 2 torr therein , and providing a working voltage of 5 kev , so as to make the dry first powder composed of the thrombin factor adsorb to calcium sulfate hemihydrate via the physical electronic - adsorption action ; ( 3 ) providing the reaction container ( made of acrylic material ) having at least one recessed space with a length of 5 cm , a width of 2 cm and a depth of 2 cm ; ( 4 ) providing medical - grade injection water and a stirrer , mixing the medical - grade injection water respectively with second powder ( rich - contained with the growth factor ) and pure calcium sulfate hemihydrate in the mixing proportion of 2 : 1 to get a first mixture and a second mixture , putting the first mixture and the second mixture into the recessed space of the reaction container , and stirring the stirrer in a rate of 30 revolution per minute ( rpm ) for 15 seconds to make the first mixture and second mixture within the recessed space be sufficiently mixed ; and ( 5 ) preparing the vicat needle , with a tip area of 1 mm 2 and a weight of 300 g , served as test equipment , laying the tip of the vicat needle on the first mixture and second mixture in each interval of 30 seconds until no recessed mark appears on the surface of the first mixture and second mixture , and respectively recording the how many intervals spent until no recessed mark appears on the surface of the first mixture and second mixture , so as to respectively get a first hardening time and a second hardening time . summarizing above description , the method for manufacturing the biomedical filler materials , which are rich - contained with the thrombin factor and the growth factor as provided in the present invention , has the advantages of : ( 1 ) adsorbing the dry first powder contained with the thrombin to the surface of the filler materials via the physical electro - adsorption action ; ( 2 ) the filler materials are characterized in having an appearance with power composed of particles with different sizes ; as time going , to form biomedical filler materials , which have specified strengths after mixing with different mixed - liquids ; and ( 3 ) it is not necessary to do complicated mixing procedure when an operation is in progress , so as to make the operation can be executed more quickly and decrease the risk of the failure of the operation . although the present invention has been described with reference to the preferred embodiments thereof , it is apparent to those skilled in the art that a variety of modifications and changes may be made without departing from the scope of the present invention which is intended to be defined by the appended claims .

a method for manufacturing at least one kind of biomedical filler materials rich - contained with at least one growth factor is disclosed , wherein at least one dry powder composed of the growth factor is provided to cover the surface of a plurality of filler materials via a physical electro - adsorption action . the filler materials have different intensities of electrostatic charge on their surfaces ; therefore , with different values of electrovalence , the dry powder can be adsorbed to the surface of the filler materials via the physical electro - adsorption action , so as to obtain the biomedical filler materials rich - contained with the thrombin and the growth factor .

with reference to fig1 and 2 , the autoinjector device according to the invention comprises an outer housing 1 , having a tubular shape and equipped with a cap - shaped knob 3 at one end , while a tapered end of a trigger slide 30 , internal to the outer housing 1 , projects from the other end . the tapered end is closed by a needle cap 41 and an inner needle shield 42 sitting directly over the needle is housed in the needle cap 41 . needle cap 41 and needle shield 42 are easily removable when the device is to be used in order to expose the needle ready for the first dose injection . the autoinjector is further provided with a protective sheath 40 sitting over the outer housing 1 when the device is in the storage state . with reference also to fig3 and 4 , the outer housing 1 extends over a longitudinal axis x between an open ( or distal ) end and a closed ( or proximal ) end , having a closure wall la , on which two diametrically opposed , arc grooves 2 ( only one shown in fig5 ) are formed each having an angular extension of , for example , about 90 °. the outer housing 1 further has an end portion 1 b at its closed end with a lower diameter and defining a surface acting as a seat for the cap - shaped knob 3 . two pins 4 engaging with grooves 2 extend from the bottom side of the knob 3 . the knob 3 is mounted coaxially to the outer housing 1 and is pivotable on the end portion 1 b of the outer housing 1 and the extent of its clockwise or anticlockwise , angular displacement depends on the angular extension of the grooves 2 . a rod 3 a extends from knob 3 parallel to longitudinal axis x along the outer housing 1 . rod 3 a serves as pointer to indicate the actual operating condition of the autoinjector device ( stored state , first triggerable state , i . e . ready for the first injection , second triggerable state , i . e . ready for the second injection ) in combination to as many corresponding reference marks 5 ( for instance consisting of reference numerals 0 , 1 , 2 ) arranged in a circumferential line on the surface of outer housing 1 . a portion 1 c of the outer housing 1 , extending from its open end , is formed with a greater inner diameter than the remaining part of the outer housing 1 , whereby a circular step 6 is formed between the two parts . two diametrically opposed slots 7 are formed on the same portion 1 c close to the step 6 , while two further diametrically opposed slots 8 are formed near the open end of the outer housing 1 . the side wall of the end portion 1 b of the outer housing 1 is formed with two diametrically opposed side grooves 9 and two tabs 10 projecting from the side surface of a tubular member 11 engage therewith . tubular member 11 is open at its distal end and closed by a wall 11 a at its proximal end . with reference also to fig5 , 8 a and 8 b , the tubular member 11 is coaxially engaged within the tubular outer housing 1 and wall 11 a is formed with two holes 12 for a fixed connection with the ends of the two knob pins 4 passing through the arc grooves 2 , whereby the tubular member 11 is integral to knob 3 and can angularly displace with it but cannot axially slide due to the bent - out tabs 10 engaging with the side grooves 9 of the outer housing 1 . two diametrically opposed , circumferential slots 13 are formed on the lateral surface of the tubular member 11 at an intermediate position and respective through guides 14 extend diagonally along the side surface of the tubular member 11 toward the distal end thereof , each with an angular extension of about 90 °. the two through guides 14 have equal slope , but are symmetrically opposed relative to longitudinal axis x . the two through guides 14 are formed each by a first and a second inclined tract 14 a and 14 b separated by an intermediate step 14 c lying on a plane orthogonal to the longitudinal axis x and an end tract 14 d extending longitudinally from the end of the second tract 14 b to the open end of the tubular member 11 . the first inclined tract 14 a of the through guides 14 starts from an intermediate position of the respective circumferential slots 13 and defines in this way two slot parts 13 a and 13 b placed at the right side and at the left side of the first tract 14 a . a flap 15 , the function of which will be explained later on , extends out from the lower side of the slot part 13 b close to the first tract 14 a . an inner housing 16 of tubular shape with open ends , shown in detail in fig1 a , 10 b and 10 c , is coaxially housed in the tubular member 11 . two diametrically opposed , longitudinal grooves 17 are formed on the inner housing 16 and at least a longitudinal cut 18 , serving as viewing window for the drug extends from the distal end thereof . two radial teeth 19 extend outwardly from the inner housing 16 for engaging within the slots 7 of the outer housing 1 , preventing the inner housing 16 from moving linearly and rotationally relative to the outer housing 1 . a pair of radial projections 20 extend from inner housing 16 to rest against the step 6 ( see fig4 ) of the outer housing 1 to further prevent any axial movement . with reference to fig6 , a syringe assembly s is shown therein . the assembly comprises a syringe 21 with a needle 22 at its distal end . the syringe is housed in a tubular support 23 , also shown in fig9 , open at its ends and having an intermediate enlargement 23 c fitting against the distal end of the inner housing 16 , wherein the tubular support 23 is partly housed . a window 23 d is formed on tubular support 23 and is aligned to longitudinal cut 18 of inner housing 16 to make the syringe content visible from the outside . the tubular support 23 is formed with bent - in edges 23 a at its distal end against which the syringe distal end abuts , while two opposed radial projections 23 b are formed at the other end on which a flange 21 a of the syringe 21 rests . the two radial projections 23 b of the tubular support 23 are further configured to be slidably engaged within the longitudinal grooves 17 of the inner housing 16 to allow the tubular support 23 to axially move for a set extent , whereby the syringe housed therewith is correspondingly moved . two inclined guides 24 are formed on the wall of the inner housing 16 at diametrically opposed parts , said guides being inclined relative to the longitudinal axis x , but not incident thereto , because they are formed on the lateral surface of the inner housing 16 . in particular , as shown in fig1 a and 10 b , each inclined guide 24 comprises a first and a second ramp 24 a and 24 b separated by an intermediate land 24 c and ending with two end lands 24 d and 24 e formed at the beginning of the first ramp 24 a and , respectively , at the end of the second ramp 24 b . the first ramp 24 a has a length greater than the second ramp 24 b . the inclined guides 24 have equal inclination and a substantially equal extension of the through guides 14 of the tubular member 11 . inside the syringe 21 a plunger stopper 25 is slidably and sealingly housed and is connected to the end of a plunger 26 axially sliding within the inner housing 16 . in particular the plunger 26 has an enlarged intermediate portion 26 a of a substantially cylindrical shape and diameter substantially equal to the inner diameter of the inner housing 16 in order to keep the plunger in axial alignment to the inner housing 16 when sliding therein . as shown in fig6 , at the end opposed to that bearing the stopper 25 the plunger 26 has an axial stem 26 b with an injection spring 27 arranged around it and placed between the wall 11 a of the tubular member 11 and a cam plate 28 coaxially and slidably mounted on the stem 26 b and placed close to the enlargement 26 a of the plunger 26 . the injection spring 27 is mounted in a compressed state between the wall 11 a and the cam plate 28 and therefore is pre - loaded when the device is in the rest or storage condition . two slide pins 29 extend radially from cam plate 28 at diametrically opposed sides thereof and are configured to rest on flaps 15 of the tubular member 11 when the device is in the rest or storage condition and to be able to slide in the respective inclined guide 24 of the inner housing 16 . the inner housing 16 , which houses the syringe support 23 and the syringe 21 therein , both slidable in the axial direction , is arranged in the trigger slide 30 that is shaped with the tapered end from which the syringe needle 22 extends when the device is used . the syringe support 23 projects from the inner housing 16 starting from its intermediate enlargement 23 c , which has an essentially cylindrical shape and a diameter substantially equal to the inner diameter of the trigger slide 30 to allow for the relative sliding while keeping the axial alignment . a push - back spring 31 is placed between the intermediate enlargement 23 c of the syringe support 23 and the tapered end of the trigger slide 30 in order to keep the syringe support 23 biased against the end of the inner housing 16 . from the trigger slide 30 , near its tapered end , there extend two diametrical teeth 32 ( only one visible in fig1 ) which engage slidably in the slots 8 of the outer housing 1 , whereby the trigger slide 30 is enabled to slide axially over a distance equal to the length of the slots 8 relative to the outer housing 1 . the other end ( the proximal end ) of the trigger slide 30 , which has a substantially tubular shape , is arranged between the outer housing 1 and the tubular member 11 and is configured in the shape of legs , which , in the present embodiment of the invention , are formed with sloped profile having substantially the same inclination as that of the inclined guides 24 formed on the inner housing 16 . in particular , as shown in fig7 , starting from said end the trigger slide 30 is formed with two diametrically opposed , longitudinal cuts 30 a defining two portions of tubular surface each delimited by a first side coinciding with a generatrix of the tubular trigger slide and a second side inclined relative to said generatrix . the inclined side of each tubular surface portion is formed by a long leg 36 , starting from the proximal end of the trigger slide 30 , and a short leg 37 separated by a recess 38 . when the autoinjector is operated , the slide pins 29 of cam plate 28 coaxial to stem 26 b of plunger 26 slidingly engage , as said before , in the respective inclined guides 24 of the inner housing 16 passing through the through guides 14 of the tubular member 11 and abutting against the inclined profiles of the respective long and short legs 36 and 37 of the trigger slide 30 . in summary , knob 3 and tubular member 11 are integral to each other and pivotable relative to the outer housing 1 and the tubular member 11 is unable to axially slide relative thereto . the inner housing 16 is fixed relative to the outer housing 1 and therefore is unable to move linearly and rotationally relative thereto . the trigger slide 30 is axially slidable relative to the outer housing 1 , but cannot rotate relative thereto , and the syringe support 23 is slidable axially in the inner housing 16 and in the trigger slide 30 against the push - back spring 31 . the plunger 26 is axially slidable in the inner housing 16 under the action of the injection spring 27 once it is triggered . since the inner housing 16 is fixed to the outer housing 1 , the sliding of the slide pins 29 on the inclined profiles of the legs 36 and 37 and within the inclined guides 24 of the inner housing 16 causes the syringe support 23 and the plunger 26 to axially move . the operation of the autoinjector device according to the invention is now described with reference to fig1 to 22 . in the stored condition , the injection spring 27 rests in a fully compressed state between the wall 11 a of the tubular member 11 and the cam plate 28 , as shown in fig3 . the slide pins 29 of the cam plate 28 rest on the bent - out flaps 15 of the tubular member 11 , thereby securing the injection spring 27 in place ( fig1 ). this to prevent creep of the mechanism over a medium to long - term storage period . the stem 26 a of the plunger 26 also helps to prevent the compressed injection spring 27 from deflecting . the tubular member 11 is connected to the outer housing 1 through the projecting tabs 10 clipped in the grooves 9 to allow the rotation and prevent the translation of the tubular member 11 with respect to the outer housing 1 , when the knob 3 is rotated . the inner housing 16 sits in the tubular member 11 and is fixed to the outer housing 1 by the teeth 19 and projections 20 preventing them from rotating and translating relative to each other . the support 23 of the syringe 21 is seated in the trigger slide 30 and slidingly engages its radial projections 23 b in the longitudinal grooves 17 of the inner housing 16 . the length of these grooves defines how far the syringe support 23 can move which ultimately sets the needle extension distance . once the protective sheath 40 is removed , to perform the first automatic injection the user must remove the needle cap 41 from the tapered end of the trigger slide 30 . in this step the needle is uncovered , but remains sub - flush within the tapered end of the trigger slide . to arm the device for the first dose , the user must rotate the cap - shaped knob 3 by a set angle . twisting the knob 3 from the position 0 ( stored state ) to position 1 ( first dose armed state ) causes the tubular member 11 to rotate a corresponding set angle , as it is integrally linked to the knob 3 ( fig1 ), and the alignment of the first inclined tract 14 a of the through guides 14 to the first ramp 24 a of the inclined guides 24 . this action causes the slide pins 29 of the cam plate 28 to drop from the bent - out flaps 15 of the tubular member 11 onto the respective first lands 24 d of the inclined guides 24 of the inner housing 16 , thereby allowing the injection spring 27 to decompress a small set distance . in this step no other components change position or orientation . the device is now armed and ready to be triggered by the user . the device is triggered by the user pressing the tapered end of the trigger slide 30 against the injection site ( fig1 ). depressing the trigger slide 30 forces the component to move inside the outer housing 1 towards the opposite end . this causes the ends of the long legs 36 of the trigger slide 30 to exert a side force against the slide pins 29 of the cam plate 28 , which rotates around stem 26 b to arrange the pins 29 into alignment to the first ramp 24 a of the inclined guides 24 of the inner housing 16 ( fig1 ), whereby these pins 29 are free to slide in the first ramp 24 a under the action of the injection spring 27 . once triggered , the injection spring 27 is free to decompress and thereby forces the cam plate 28 , the plunger 26 and the syringe 21 forward , i . e . toward the distal end of the outer housing 1 ( fig1 ). this movement also causes the syringe needle 22 to enter the injection site by a depth governed by the length of the longitudinal grooves 17 guiding the syringe support 23 on the inner housing 16 . in order that this movement can occur , the hydrostatic force required to expel the medicament from the syringe needle must be greater than the sum of the frictional forces to push the syringe assembly s forward to the specified needle injection depth . once the syringe needle 22 reaches the full injection depth ( fig1 ), the injection spring 27 continues to decompress pushing the cam plate 28 and the plunger 26 forward with respect to the syringe barrel and causing the first dose of medicament to be expelled from the syringe . it is worth noting that the cam plate 28 , while axially moving , rotates around the stem 26 a of the plunger 26 , as the slide pins 29 are constrained to slide in the inclined guides 24 . it is also worth noting that the length of the ramp 24 a of the inclined guides 24 is such as to allow both the sliding of the syringe support 23 , to cause the needle to penetrate the skin , and the sliding of the plunger 26 to delivery the first drug dose . the first dose delivery is complete once the slide pins 29 reach the intermediate land 24 c of the inclined guides 24 on the inner housing 16 and the recess 38 of the trigger slide 30 ( fig1 ). as the land 24 c prevents the injection spring 27 from decompressing any further and the engagement in the recess 38 prevents the slide pins 29 from rotating any further , no more medicament will be expelled . after delivery of the first dose , the user removes the device from the injection site . since the pressure on the tapered end of the trigger slide 30 ceases , the push - back spring 31 moves forward the tapered end reducing the exposed needle length . due to this sliding of the trigger slide 30 , the slide pins 29 come out from the respective recesses 38 , while being stopped on lands 24 c of the guide 24 , as shown in fig1 . then the user re - sheaths the device in the protective sheath 40 to avoid any needle damaging or injuries and waits for a prescribed time to decide if a second dose is necessary . should a second dose be required , the user must first remove the protective sheath 40 from the device in order to access the needle and then arm the device to prepare it for the second dose delivery . to this purpose the user must rotate the cap - shaped knob 3 by a prefixed angle . rotating the knob 3 from position 1 to position 2 ( second dose injecting armed state ) causes the tubular member 11 to rotate a set angle . in this way the second inclined tract 14 b of the through guides 14 is aligned to the ramp 24 b of the inclined guides 24 and the slide pins 29 move along the intermediate land 24 c up to reach a position ready to be triggered ( fig1 ). the device is triggered by the user pressing the tapered end of the trigger slide 30 against the injection site and causing the trigger slide to move toward the opposite end of the outer housing 1 . the end of short legs 37 of the trigger slide 30 comes into contact with the respective slide pin 29 and displaces it laterally on the intermediate land 24 c up to bring it into alignment to the second ramp 24 b of the inclined guide 24 on the inner housing 16 , thus allowing the slide pins 29 to freely move in the respective second ramps 24 b ( fig2 ) under the action of the injection spring 27 which depresses further . in this way a further sliding of the cam plate 28 and the piston 26 relative to the syringe barrel is caused to expel the medicament from the syringe , i . e . the delivery of the second dose . it is worth noting that , since the needle remains exposed from the first dose , there is no need to push the syringe assembly forward and therefore a lower length of the second ramp 24 b with respect to the first ramp 24 a is necessary . the delivery of the second dose is complete once the slide pins 29 reach the end land 24 e of the inclined guides 24 on the inner housing 16 ( fig2 ). as the end land 24 e prevents the injection spring 27 from decompressing any further , no more medicament will be expelled . after delivery of the second dose , the user removes the device from the injection site . as the pressure on the tapered end of the trigger slide 30 ceases , the push - back spring 31 moves forward the tapered end ( fig2 ). finally , the user re - sheaths the device for its safe disposal . the tubular member 11 is preferably made of metallic material to provide adequate protection to the syringe and to ensure a long - term storage of the device , as it is not prone to creep to the same extent that the other moulded device components would be . the autoinjector device according to the invention is suitable to the delivery of medicaments in solution , especially epinephrine ( also known as adrenaline ). in particular , the doses of epinephrine that can be administered with the device of the invention are preferably in the range of 0 . 05 mg to 0 . 5 mg for each delivered dose ( from 0 . 1 mg to 1 mg if two doses are considered ). preferred doses for each delivery are 0 . 05 mg , 0 . 10 mg , 0 . 15 mg , 0 . 30 mg and 0 . 50 mg . the above doses are based on a concentration of the epinephrine solution preferably ranging from 0 . 05 mg / ml to 0 . 5 mg / ml , the concentrations 0 . 05 mg / ml , 0 . 1 mg / ml , 0 . 16 mg / ml , 0 . 3 mg / ml and 0 . 5 mg / ml being particularly preferred .

a two - dose autoinjector for a medicament wherein the locking and releasing of the drive spring of the autoinjector is controlled through stepped guides with ramps for two successive slidings of slides operated by the spring and connected with the syringe and plunger . the guides and the slides are pivotable relative to one another and the sliding direction , while the syringe can only slide axially . to enable or disable the sliding of the slides within the guides an angularly angularly mobile arming member is provided formed with a guide track substantially equal to that of the stationary member where the guides are formed .

the novel features of the invention are set forth with particularity in the appended claims . a better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments , in which the principles of the invention are utilized . while preferred embodiments of the present invention have been shown and described herein such embodiments are provided by way of example only . it should be understood that various alternatives to the embodiments of the invention described herein may be employed in practicing the invention . those ordinary skilled in the art will appreciate that numerous variations , changes , and substitutions are possible without departing from the invention . it is intended that the following claims define the scope of aspects of the invention and that methods and structures within the scope of these claims and their equivalents be covered thereby . the section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described . all documents , or portions of documents , cited in the application including , without limitation , patents , patent applications , articles , books , manuals , and treatises are hereby expressly incorporated by reference in their entirety for any purpose . unless defined otherwise , all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs . all patents , patent applications , published materials referred to throughout the entire disclosure herein , unless noted otherwise , are incorporated by reference in their entirety . in the event that there is a plurality of definitions for terms herein , those in this section prevail . where reference is made to a url or other such identifier or address , it is understood that such identifiers can change and particular information on the internet can come and go , but equivalent information can be found by searching the internet or other appropriate reference source . reference thereto evidences the availability and public dissemination of such information . it is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed . in this application , the use of the singular includes the plural unless specifically stated otherwise . it must be noted that , as used in the specification and the appended claims , the singular forms “ a ”, “ an ” and “ the ” include plural referents unless the context clearly dictates otherwise . it should also be noted that use of “ or ” means “ and / or ” unless stated otherwise . furthermore , use of the term “ including ” as well as other forms , such as “ include ”, “ includes ”, and “ included ” is not limiting . likewise , use of the term “ comprising ” as well as other forms , such as “ comprise ”, “ comprises ”, and “ comprised ” is not limiting . definition of standard chemistry terms may be found in reference works , including carey and sundberg “ advanced organic chemistry 4 th ed .” vols . a ( 2000 ) and b ( 2001 ), plenum press , new york . unless otherwise indicated , conventional methods of mass spectroscopy , nmr , hplc , ir and uv / vis spectroscopy and pharmacology , within the skill of the art are employed . unless specific definitions are provided , the nomenclature employed in connection with , and the laboratory procedures and techniques of , analytical chemistry , synthetic organic chemistry , and medicinal and pharmaceutical chemistry described herein are those known in the art . standard techniques can be used for chemical syntheses , chemical analyses , pharmaceutical preparation , formulation , and delivery , and treatment of patients . reactions and purification techniques can be performed e . g ., using kits of manufacturer &# 39 ; s specifications or as commonly accomplished in the art or as described herein . the foregoing techniques and procedures can be generally performed of conventional methods well known in the art and as described in various general and more specific references that are cited and discussed throughout the present specification . throughout the specification , groups and substituents thereof can be chosen by one skilled in the field to provide stable moieties and compounds . where substituent groups are specified by their conventional chemical formulas , written from left to right , they equally encompass the chemically identical substituents that would result from writing the structure from right to left . as a non - limiting example , ch 2 o is equivalent to och 2 . unless otherwise noted , the use of general chemical terms , such as though not limited to “ alkyl ,” “ amine ,” “ aryl ,” are equivalent to their optionally substituted forms . for example , “ alkyl ,” as used herein , includes optionally substituted alkyl . the compounds presented herein may possess one or more stereocenters and each center may exist in the r or s configuration , or combinations thereof . likewise , the compounds presented herein may possess one or more double bonds and each may exist in the e ( trans ) or z ( cis ) configuration , or combinations thereof . presentation of one particular stereoisomer , regioisomer , diastereomer , enantiomer or epimer should be understood to include all possible stereoisomers , regioisomers , diastereomers , enantiomers or epimers and mixtures thereof . thus , the compounds presented herein include all separate configurational stereoisomeric , regioisomeric , diastereomeric , enantiomeric , and epimeric forms as well as the corresponding mixtures thereof . techniques for inverting or leaving unchanged a particular stereocenter , and those for resolving mixtures of stereoisomers are well known in the art and it is well within the ability of one of skill in the art to choose an appropriate method for a particular situation . see , for example , fumiss et al . ( eds . ), vogel &# 39 ; s encyclopedia of practical organic chemistry 5 . sup . th ed ., longman scientific and technical ltd ., essex , 1991 , 809 - 816 ; and heller , acc . chem . res . 1990 , 23 , 128 . the terms “ moiety ”, “ chemical moiety ”, “ group ” and “ chemical group ”, as used herein refer to a specific segment or functional group of a molecule . chemical moieties are often recognized chemical entities embedded in or appended to a molecule . the term “ bond ” or “ single bond ” refers to a chemical bond between two atoms , or two moieties when the atoms joined by the bond are considered to be part of larger substructure . the term “ catalytic group ” refers to a chemical functional group that assists catalysis by acting to lower the activation barrier to reaction . the term “ optional ” or “ optionally ” means that the subsequently described event or circumstance may or may not occur , and that the description includes instances where said event or circumstance occurs and instances in which it does not . for example , “ optionally substituted alkyl ” means either “ alkyl ” or “ substituted alkyl ” as defined below . further , an optionally substituted group may be un - substituted ( e . g ., ch 2 ch 3 ), fully substituted ( e . g ., cf 2 cf 3 ), mono - substituted ( e . g ., ch 2 ch 2 f ) or substituted at a level anywhere in - between fully substituted and mono - substituted ( e . g ., ch 2 chf 2 , cf 2 ch 3 , cfhchf 2 , etc ). it will be understood by those skilled in the art with respect to any group containing one or more substituents that such groups are not intended to introduce any substitution or substitution patterns ( e . g ., substituted alkyl includes optionally substituted cycloalkyl groups , which in turn are defined as including optionally substituted alkyl groups , potentially ad infinitum ) that are sterically impractical and / or synthetically non - feasible . thus , any substituents described should generally be understood as having a maximum molecular weight of about 1 , 000 daltons , and more typically , up to about 500 daltons ( except in those instances where macromolecular substituents are clearly intended , e . g ., polypeptides , polysaccharides , polyethylene glycols , dna , rna and the like ). as used herein , c 1 - cn , includes c 1 - c 2 , c 1 - c 3 . . . c 1 - cn . by way of example only , a group designated as “ c 1 - c 4 ” indicates that there are one to four carbon atoms in the moiety , i . e . groups containing 1 carbon atom , 2 carbon atoms , 3 carbon atoms or 4 carbon atoms , as well as the ranges c 1 - c 2 and c 1 - c 3 . thus , by way of example only , “ c 1 - c 4 alkyl ” indicates that there are one to four carbon atoms in the alkyl group , i . e ., the alkyl group is selected from among methyl , ethyl , propyl , iso - propyl , n - butyl , isobutyl , sec - butyl , and t - butyl . whenever it appears herein , a numerical range such as “ 1 to 10 ” refers to each integer in the given range ; e . g ., “ 1 to 10 carbon atoms ” means that the group may have 1 carbon atom , 2 carbon atoms , 3 carbon atoms , 4 carbon atoms , 5 carbon atoms , 6 carbon atoms , 7 carbon atoms , 8 carbon atoms , 9 carbon atoms , or 10 carbon atoms . the term “ hydrocarbon ” as used herein , alone or in combination , refers to a compound or chemical group containing only carbon and hydrogen atoms . the terms “ heteroatom ” or “ hetero ” as used herein , alone or in combination , refer to an atom other than carbon and hydrogen . heteroatoms are independently selected from among oxygen , nitrogen , sulfur , phosphorous , silicon , selenium and tin but are not limited to these atoms . in embodiments in which two or more heteroatoms are present , the two or more heteroatoms can be the same as each another , or some or all of the two or more heteroatoms can each be different from the others . the term “ alkyl ” as used herein , alone or in combination , refers to an optionally substituted straight - chain , or optionally substituted branched - chain saturated hydrocarbon monoradical having from one to about ten carbon atoms , more preferably one to six carbon atoms . examples include , but are not limited to methyl , ethyl , n - propyl , isopropyl , 2 - methyl - 1 - propyl , 2 - methyl - 2 - propyl , 2 - methyl - 1 - butyl , 3 - methyl - 1 - butyl , 2 - methyl - 3 - butyl , 2 , 2 - dimethyl - 1 - propyl , 2 - methyl - 1 - pentyl , 3 - methyl - 1 - pentyl , 4 - methyl - 1 - pentyl , 2 - methyl - 2 - pentyl , 3 - methyl - 2 - pentyl , 4 - methyl - 2 - pentyl , 2 , 2 - dimethyl - 1 - butyl , 3 , 3 - dimethyl - 1 - butyl , 2 - ethyl - 1 - butyl , n - butyl , isobutyl , sec - butyl , t - butyl , n - pentyl , isopentyl , neopentyl , tert - amyl and hexyl , and longer alkyl groups , such as heptyl , octyl and the like . whenever it appears herein , a numerical range such as “ c 1 - c 6 alkyl ” or “ c 1 - 6 alkyl ”, means that the alkyl group may consist of 1 carbon atom , 2 carbon atoms , 3 carbon atoms , 4 carbon atoms , 5 carbon atoms or 6 carbon atoms , although the present definition also covers the occurrence of the term “ alkyl ” where no numerical range is designated . the term “ alkylene ” as used herein , alone or in combination , refers to a diradical derived from the above - defined monoradical , alkyl . examples include , but are not limited to methylene (— ch 2 ), ethylene (— ch 2 ch 2 ), propylene (— ch 2 ch 2 ch 2 ), isopropylene (— ch ( ch 3 ) ch 2 ) and the like . the term “ alkenyl ” as used herein , alone or in combination , refers to an optionally substituted straight - chain , or optionally substituted branched - chain hydrocarbon monoradical having one or more carbon - carbon double - bonds and having from two to about ten carbon atoms , more preferably two to about six carbon atoms . the group may be in either the cis or trans conformation about the double bond ( s ), and should be understood to include both isomers . examples include , but are not limited to ethenyl ( ch — ch 2 ), 1 - propenyl ( ch 2 ch ═ ch 2 ), isopropenyl [ c ( ch 3 )═ ch 2 ], butenyl , 1 , 3 - butadienyl and the like . whenever it appears herein , a numerical range such as “ c 2 - c 6 alkenyl ” or “ c 2 6 alkenyl ”, means that the alkenyl group may consist of 2 carbon atoms , 3 carbon atoms , 4 carbon atoms , 5 carbon atoms or 6 carbon atoms , although the present definition also covers the occurrence of the term “ alkenyl ” where no numerical range is designated . the term “ alkenylene ” as used herein , alone or in combination , refers to a diradical derived from the above - defined monoradical alkenyl . examples include , but are not limited to ethenylene ( ch — ch ), the propenylene isomers ( e . g ., ch 2 ch ═ ch and c ( ch 3 )═ ch ) and the like . the term “ alkynyl ” as used herein , alone or in combination , refers to an optionally substituted straight - chain or optionally substituted branched - chain hydrocarbon monoradical having one or more carbon - carbon triple - bonds and having from two to about ten carbon atoms , more preferably from two to about six carbon atoms . examples include , but are not limited to ethynyl , 2 - propynyl , 2 - butynyl , 1 , 3 - butadiynyl and the like . whenever it appears herein , a numerical range such as “ c 2 - c 6 alkynyl ” or “ c 2 - 6 alkynyl ”, means that the alkynyl group may consist of 2 carbon atoms , 3 carbon atoms , 4 carbon atoms , 5 carbon atoms or 6 carbon atoms , although the present definition also covers the occurrence of the term “ alkynyl ” where no numerical range is designated . the term “ alkynylene ” as used herein , alone or in combination , refers to a diradical derived from the above - defined monoradical , alkynyl . examples include , but are not limited to ethynylene (— cc —), propargylene (— ch 2 cc —) and the like . the term “ aliphatic ” as used herein , alone or in combination , refers to an optionally substituted , straight - chain or branched - chain , non - cyclic , saturated , partially unsaturated , or fully unsaturated nonaromatic hydrocarbon . thus , the term collectively includes alkyl , alkenyl and alkynyl groups . the terms “ heteroalkyl ”, “ heteroalkenyl ” and “ heteroalkynyl ” as used herein , alone or in combination , refer to optionally substituted alkyl , alkenyl and alkynyl structures respectively , as described above , in which one or more of the skeletal chain carbon atoms ( and any associated hydrogen atoms , as appropriate ) are each independently replaced with a heteroatom ( i . e . an atom other than carbon , such as though not limited to oxygen , nitrogen , sulfur , silicon , phosphorous , tin or combinations thereof . the terms “ haloalkyl ”, “ haloalkenyl ” and “ haloalkynyl ” as used herein , alone or in combination , refer to optionally substituted alkyl , alkenyl and alkynyl groups respectively , as defined above , in which one or more hydrogen atoms is replaced by fluorine , chlorine , bromine or iodine atoms , or combinations thereof . in some embodiments two or more hydrogen atoms may be replaced with halogen atoms that are the same as each another ( e . g . difluoromethyl ); in other embodiments two or more hydrogen atoms may be replaced with halogen atoms that are not all the same as each other ( e . g . 1 - chloro - 1 - fluoro - 1 - iodoethyl ). non - limiting examples of haloalkyl groups are fluoromethyl and bromoethyl . a non - limiting example of a haloalkenyl group is bromoethenyl . a non - limiting example of a haloalkynyl group is chloroethynyl . the term “ perhalo ” as used herein , alone or in combination , refers to groups in which all of the hydrogen atoms are replaced by fluorines , chlorines , bromines , iodines , or combinations thereof . thus , as a non - limiting example , the term “ perhaloalkyl ” refers to an alkyl group , as defined herein , in which all of the h atoms have been replaced by fluorines , chlorines , bromines or iodines , or combinations thereof . a non - limiting example of a perhaloalkyl group is bromo , chloro , fluoromethyl . a non - limiting example of a perhaloalkenyl group is trichloroethenyl . a non - limiting example of a perhaloalkynyl group is tribromopropynyl . the term “ carbon chain ” as used herein , alone or in combination , refers to any alkyl , alkenyl , alkynyl , heteroalkyl , heteroalkenyl or heteroalkynyl group , which is linear , cyclic , or any combination thereof . if the chain is part of a linker and that linker comprises one or more rings as part of the core backbone , for purposes of calculating chain length , the “ chain ” only includes those carbon atoms that compose the bottom or top of a given ring and not both , and where the top and bottom of the ring ( s ) are not equivalent in length , the shorter distance shall be used in determining the chain length . if the chain contains heteroatoms as part of the backbone , those atoms are not calculated as part of the carbon chain length . the terms “ cycle ”, “ cyclic ”, “ ring ” and “ membered ring ” as used herein , alone or in combination , refer to any covalently closed structure , including alicyclic , heterocyclic , aromatic , heteroaromatic and polycyclic fused or non - fused ring systems as described herein . rings can be optionally substituted . rings can form part of a fused ring system . the term “ membered ” is meant to denote the number of skeletal atoms that constitute the ring . thus , by way of example only , cyclohexane , pyridine , pyran and pyrimidine are six - membered rings and cyclopentane , pyrrole , tetrahydrofuran and thiophene are five - membered rings . the term “ fused ” as used herein , alone or in combination , refers to cyclic structures in which two or more rings share one or more bonds . the term “ aromatic ” as used herein , refers to a planar , cyclic or polycyclic , ring moiety having a delocalized at - electron system containing 4n + 2n electrons , where n is an integer . aromatic rings can be formed by five , six , seven , eight , nine , or more than nine atoms . aromatics can be optionally substituted and can be monocyclic or fused - ring polycyclic . the term aromatic encompasses both all carbon containing rings ( e . g ., phenyl ) and those rings containing one or more heteroatoms ( e . g ., pyridine ). the term “ aryl ” as used herein , alone or in combination , refers to an optionally substituted aromatic hydrocarbon radical of six to about twenty ring carbon atoms , and includes fused and non - fused aryl rings . a fused aryl ring radical contains from two to four fused rings where the ring of attachment is an aryl ring , and the other individual rings may be alicyclic , heterocyclic , aromatic , heteroaromatic or any combination thereof . further , the term aryl includes fused and non - fused rings containing from six to about twelve ring carbon atoms , as well as those containing from six to about ten ring carbon atoms . a non - limiting example of a single ring aryl group includes phenyl ; a fused ring aryl group includes naphthyl , phenanthrenyl , anthracenyl , azulenyl ; and a non - fused bi - aryl group includes biphenyl . the term “ arylene ” as used herein , alone or in combination , refers to a diradical derived from the above - defined monoradical , aryl . examples include , but are not limited to 1 , 2 - phenylene , 1 , 3 - phenylene , 1 , 4 - phenylene , 1 , 2 - naphthylene and the like . the term “ heteroaryl ” as used herein , alone or in combination , refers to optionally substituted aromatic mono - radicals containing from about five to about twenty skeletal ring atoms , where one or more of the ring atoms is a heteroatom independently selected from among oxygen , nitrogen , sulfur , phosphorous , silicon , selenium and tin but not limited to these atoms and with the proviso that the ring of said group does not contain two adjacent o or s atoms . in embodiments in which two or more heteroatoms are present in the ring , the two or more heteroatoms can be the same as each another , or some or all of the two or more heteroatoms can each be different from the others . the term heteroaryl includes optionally substituted fused and non - fused heteroaryl radicals having at least one heteroatom . the term heteroaryl also includes fused and non - fused heteroaryls having from five to about twelve skeletal ring atoms , as well as those having from five to about ten skeletal ring atoms . bonding to a heteroaryl group can be via a carbon atom or a heteroatom . thus , as a non - limiting example , an imidiazole group may be attached to a parent molecule via any of its carbon atoms ( imidazol - 2 - yl , imidazol - 4 - yl or imidazol - 5 - yl ), or its nitrogen atoms ( imidazol - 1 - yl or imidazol - 3 - yl ). likewise , a heteroaryl group may be further substituted via any or all of its carbon atoms , and / or any or all of its heteroatoms . a fused heteroaryl radical may contain from two to four fused rings where the ring of attachment is a heteroaromatic ring and the other individual rings may be alicyclic , heterocyclic , aromatic , heteroaromatic or any combination thereof . anon - limiting example of a single ring heteroaryl group includes pyridyl ; fused ring heteroaryl groups include benzimidazolyl , quinolinyl , acridinyl ; and a non - fused bi - heteroaryl group includes bipyridinyl . further examples of heteroaryls include , without limitation , furanyl , thienyl , oxazolyl , acridinyl , phenazinyl , benzimidazolyl , benzofuranyl , benzoxazolyl , benzothiazolyl , benzothiadiazolyl , benzothiophenyl , benzoxadiazolyl , benzotriazolyl , imidazolyl , indolyl , isoxazolyl , isoquinolinyl , indolizinyl , isothiazolyl , isoindolyloxadiazolyl , indazolyl , pyridyl , pyridazyl , pyrimidyl , pyrazinyl , pyrrolyl , pyrazolyl , purinyl , phthalazinyl , pteridinyl , quinolinyl , quinazolinyl , quinoxalinyl , triazolyl , tetrazolyl , thiazolyl , triazinyl , thiadiazolyl and the like , and their oxides , such as for example pyridyl - n - oxide and the like . the term “ heteroarylene ” as used herein , alone or in combination , refers to a diradical derived from the above - defined monoradical heteroaryl . examples include , but are not limited to pyridinylene and pyrimidinylene . the term “ heterocyclyl ” as used herein , alone or in combination , refers collectively to heteroalicyclyl and heteroaryl groups . herein , whenever the number of carbon atoms in a heterocycle is indicated ( e . g ., c 1 - c 6 heterocycle ), at least one non - carbon atom ( the heteroatom ) must be present in the ring . designations such as “ c 1 - c 6 heterocycle ” refer only to the number of carbon atoms in the ring and do not refer to the total number of atoms in the ring . designations such as “ 4 - 6 membered heterocycle ” refer to the total number of atoms that are contained in the ring ( i . e ., a four , five , or six membered ring , in which at least one atom is a carbon atom , at least one atom is a heteroatom and the remaining two to four atoms are either carbon atoms or heteroatoms ). for heterocycles having two or more heteroatoms , those two or more heteroatoms can be the same or different from one another . heterocycles can be optionally substituted . non - aromatic heterocyclic groups include groups having only three atoms in the ring , while aromatic heterocyclic groups must have at least five atoms in the ring . bonding ( i . e . attachment to a parent molecule or further substitution ) to a heterocycle can be via a heteroatom or a carbon atom . a non - limiting example of “ heterocyclyl ” includes azinyl , azetidinyl , oxetanyl , thietanyl , homopiperidinyl , oxepanyl , thiepanyl , oxazepinyl , diazepinyl , thiazepinyl , 1 , 2 , 3 , 6 - tetrahydropyridinyl , 2 - pyrrolinyl , 3 - pyrrolinyl , indolinyl , 2h - pyranyl , 4h - pyranyl , dioxanyl , 1 , 3 - dioxolanyl , pyrazolinyl , dithianyl , dithiolanyl , dihydropyranyl , dihydrothienyl , dihydrofuranyl , pyrazolidinyl , imidazolinyl , imidazolidinyl , 3 - azabicyclo [ 3 . 1 . 0 ] hexyl , 3 - azabicyclo [ 4 . 1 . 0 ] heptyl , 3h - indolyl and quinolizinyl and the like . the terms also include all ring forms of the carbohydrates , including but not limited to the monosaccharides , the disaccharides and the oligosaccharides . the term “ carbocyclyl ” as used herein , alone or in combination , refers collectively to alicyclyl and aryl groups ; i . e . all carbon , covalently closed ring structures , which may be saturated ( i . e ., cycloalkyl ), partially unsaturated ( cycloalkenyl ), fully unsaturated or aromatic . carbocyclic rings can be formed by three , four , five , six , seven , eight , nine , or more than nine carbon atoms . carbocycles can be optionally substituted . the term distinguishes carbocyclic from heterocyclic rings in which the ring backbone contains at least one atom which is different from carbon . the term “ cycloalkyl ” as used herein , alone or in combination , refers to an optionally substituted , saturated , hydrocarbon monoradical ring , containing from three to about fifteen ring carbon atoms or from three to about ten ring carbon atoms , though may include additional , non - ring carbon atoms as substituents ( e . g . methylcyclopropyl ). the terms “ halogen ”, “ halo ” or “ halide ” as used herein , alone or in combination refer to fluoro , chloro , bromo and iodo . the term “ alkoxy ” as used herein , alone or in combination , refers to an alkyl ether radical , o - alkyl , including the groups o - aliphatic and o - carbocyclyl , wherein the alkyl , aliphatic and carbocyclyl groups may be optionally substituted , and wherein the terms alkyl , aliphatic and carbocyclyl are as defined herein . non - limiting examples of alkoxy radicals include methoxy , ethoxy , n - propoxy , isopropoxy , n - butoxy , iso - butoxy , sec - butoxy , tertbutoxy and the like . the term “ alkylthiol ” as used herein , alone or in combination , refers to an alkyl sufide radical , — s - alkyl , including — s - aliphatic and — s - carbocyclyl , wherein the alkyl , aliphatic and carbocyclyl groups may be optionally substituted , and wherein the terms alkyl , aliphatic and carbocyclyl are as defined herein . preferably , the alkyl has 1 to about 6 , 1 to about 4 carbon atoms . non - limiting examples of alkylthiol radicals include methylthiol , ethylthiol , and the like . the term “ sulfinyl ” as used herein , alone or in combination , refers to the diradical — s (— o ). the term “ sulfonyl ” as used herein , alone or in combination , refers to the diradical — s (— o ) 2 . the terms “ sulfonamide ”, “ sulfonamido ” and “ sulfonamidyl ” as used herein , alone or in combination , refer to the diradical groups — s (— o ) 2 — nh — and — nh — s (═ o ) 2 . the terms “ carboxamide ” and “ carboxamido ” as used herein , alone or in combination , refer to the group of the term “ mek inhibitor ” as used herein refers to a compound that exhibits an ic 50 , with respect to mek activity , of no more than about 100 m or not more than about 50 m , as measured in the mek1 kinase assay described generally herein . “ ic 50 ” is that concentration of inhibitor which reduces the activity of an enzyme ( e . g ., mek ) to half - maximal level . compounds described herein have been discovered to exhibit inhibition against mek . compounds of the present invention preferably exhibit an ic 50 with respect to mek of no more than about 10 m , more preferably , no more than about 5 m , even more preferably not more than about 1 m , and most preferably , not more than about 200 nm , as measured in the meld kinase assay described herein . the term “ selective ,” “ selectively ,” or “ selectivity ” as used herein refers to a compound of this invention having a lower ic 50 value for a mek enzyme as compared to any other enzymes ( e . g ., at least 2 , 5 , 10 or more - fold lower ). the term may also refer to a compound of this invention having a lower ic 50 value for a mek1 enzyme as compared to a mek2 enzyme ( e . g ., at least 2 , 5 , 10 or more - fold ) or alternatively having a lower ic 50 value for a mek2 enzyme as compared to a mek1 enzyme ( e . g ., at least 2 , 5 , 10 or more - fold lower ). the term “ subject ”, “ patient ” or “ individual ” as used herein in reference to individuals suffering from a disorder , a disorder , a condition , and the like , encompasses mammals and non - mammals . examples of mammals include , but are not limited to , any member of the mammalian class : humans , non - human primates such as chimpanzees , and other apes and monkey species ; farm animals such as cattle , horses , sheep , goats , swine ; domestic animals such as rabbits , dogs , and cats ; laboratory animals including rodents , such as rats , mice and guinea pigs , and the like . examples of non - mammals include , but are not limited to , birds , fish and the like . in one embodiment of the methods and compositions provided herein , the mammal is a human . the terms “ treat ,” “ treating ” or “ treatment ,” and other grammatical equivalents as used herein , include alleviating , abating or ameliorating a disease or condition symptoms , preventing additional symptoms , ameliorating or preventing the underlying metabolic causes of symptoms , inhibiting the disease or condition , e . g ., arresting the development of the disease or condition , relieving the disease or condition , causing regression of the disease or condition , relieving a condition caused by the disease or condition , or stopping the symptoms of the disease or condition , and are intended to include prophylaxis . the terms further include achieving a therapeutic benefit and / or a prophylactic benefit . by therapeutic benefit is meant eradication or amelioration of the underlying disorder being treated . also , a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the patient , notwithstanding that the patient may still be afflicted with the underlying disorder . for prophylactic benefit , the compositions may be administered to a patient at risk of developing a particular disease , or to a patient reporting one or more of the physiological symptoms of a disease , even though a diagnosis of this disease may not have been made . the terms “ effective amount ”, “ therapeutically effective amount ” or “ pharmaceutically effective amount ” as used herein , refer to a sufficient amount of at least one agent or compound being administered which will relieve to some extent one or more of the symptoms of the disease or condition being treated . the result can be reduction and / or alleviation of the signs , symptoms , or causes of a disease , or any other desired alteration of a biological system . for example , an “ effective amount ” for therapeutic uses is the amount of the composition comprising a compound as disclosed herein required to provide a clinically significant decrease in a disease . an appropriate “ effective ” amount in any individual case may be determined using techniques , such as a dose escalation study . the terms “ administer ,” “ administering ”, “ administration ,” and the like , as used herein , refer to the methods that may be used to enable delivery of compounds or compositions to the desired site of biological action . these methods include , but are not limited to oral routes , intraduodenal mutes , parenteral injection ( including intravenous , subcutaneous , intraperitoneal , intramuscular , intravascular or infusion ), topical and rectal administration . those of skill in the art are familiar with administration techniques that can be employed with the compounds and methods described herein , e . g ., as discussed in goodman and gilman , the pharmacological basis of therapeutics , current ed . ; pergamon ; and remington &# 39 ; s , pharmaceutical sciences ( current edition ), mack publishing co ., easton , pa . in preferred embodiments , the compounds and compositions described herein are administered orally . the term “ acceptable ” as used herein , with respect to a formulation , composition or ingredient , means having no persistent detrimental effect on the general health of the subject being treated . the term “ pharmaceutically acceptable ” as used herein , refers to a material , such as a carrier or diluent , which does not abrogate the biological activity or properties of the compounds described herein , and is relatively nontoxic , i . e ., the material may be administered to an individual without causing undesirable biological effects or interacting in a deleterious manner with any of the components of the composition in which it is contained . the term “ pharmaceutical composition ,” as used herein , refers to a biologically active compound , optionally mixed with at least one pharmaceutically acceptable chemical component , such as , though not limited to carriers , stabilizers , diluents , dispersing agents , suspending agents , thickening agents , and / or excipients . the term “ carrier ” as used herein , refers to relatively nontoxic chemical compounds or agents that facilitate the incorporation of a compound into cells or tissues . the term “ agonist ,” as used herein , refers to a molecule such as a compound , a drug , an enzyme activator or a hormone modulator which enhances the activity of another molecule or the activity of a receptor site . the term “ antagonist ,” as used herein , refers to a molecule such as a compound , a drug , an enzyme inhibitor , or a hormone modulator , which diminishes , or prevents the action of another molecule or the activity of a receptor site . the term “ modulate ,” as used herein , means to interact with a target either directly or indirectly so as to alter the activity of the target , including , by way of example only , to enhance the activity of the target , to inhibit the activity of the target , to limit the activity of the target , or to extend the activity of the target . the term “ modulator ,” as used herein , refers to a molecule that interacts with a target either directly or indirectly . the interactions include , but are not limited to , the interactions of an agonist and an antagonist . the term “ pharmaceutically acceptable salt ” as used herein , refers to salts that retain the biological effectiveness of the free acids and bases of the specified compound and that are not biologically or otherwise undesirable . compounds described herein may possess acidic or basic groups and therefore may react with any of a number of inorganic or organic bases , and inorganic and organic acids , to form a pharmaceutically acceptable salt . these salts can be prepared in situ during the final isolation and purification of the compounds of the invention , or by separately reacting a purified compound in its free base form with a suitable organic or inorganic acid , and isolating the salt thus formed . examples of pharmaceutically acceptable salts include those salts prepared by reaction of the compounds described herein with a mineral or organic acid or an inorganic base , such salts including , acetate , acrylate , adipate , alginate , aspartate , benzoate , benzenesulfonate , bisulfate , bisulfite , bromide , butyrate , butyn - 1 , 4 - dioate , camphorate , camphor - sulfonate , caprylate , chlorobenzoate , chloride , citrate , cyclopentanepropionate , decanoate , digluconate , dihydrogenphosphate , dinitrobenzoate , dodecyl sulfate , ethanesulfonate , formate , fumarate , glucoheptanoate , glycerophosphate , glycolate , hemisulfate , heptanoate , hexanoate , hexyne - 1 , 6 - dioate , hydroxybenzoate , y - hydroxybutyrate , hydrochloride , hydrobromide , hydroiodide , 2 - hydroxyethanesulfonate , iodide , isobutyrate , lactate , maleate , malonate , methanesulfonate , mandelate . metaphosphate , methoxybenzoate , methylbenzoate , monohydrogenphosphate , 1 - napthalenesulfonate , 2 - napthalenesulfonate , nicotinate , nitrate , palmoate , pectinate , persulfate , 3 - phenylpropionate , phosphate , picrate , pivalate , propionate , pyrosulfate , pyrophosphate , propiolate , phthalate , phenylacetate , phenylbutyrate , propanesulfonate , salicylate , succinate , sulfate , sulfite , suberate , sebacate , sulfonate , tartrate , thiocyanate , tosylate undeconate and xylenesulfonate . other acids , such as oxalic , while not in themselves pharmaceutically acceptable , may be employed in the preparation of salts useful as intermediates in obtaining the compounds of the invention and their pharmaceutically acceptable acid addition salts ( see examples at berge et al ., j . pharm . sci . 1977 , 66 , 1 - 19 ). further , those compounds described herein which may comprise a free acid group may react with a suitable base , such as the hydroxide , carbonate or bicarbonate of a pharmaceutically acceptable metal cation , with ammonia , or with a pharmaceutically acceptable organic primary , secondary or tertiary amine . representative alkali or alkaline earth salts include the lithium , sodium , potassium , calcium , magnesium , and aluminum salts and the like . illustrative examples of bases include sodium hydroxide , potassium hydroxide , choline hydroxide , sodium carbonate , iv ′ ( c 1 _ 4 alkyl ) 4 , and the like . representative organic amines useful for the formation of base addition salts include ethylamine , diethylamine , ethylenediamine , ethanolamine , diethanolamine , piperazine and the like . it should be understood that the compounds described herein also include the quaternization of any basic nitrogen - containing groups they may contain . water or oil - soluble or dispersible products may be obtained by such quaternization . see , for example , berge et al ., supra . the term “ solvate ” as used herein refers to a combination of a compound of this invention with a solvent molecule formed by solvation . in some situations , the solvate refers to a hydrate , i . e ., the solvent molecule is a water molecule , the combination of a compound of this invention and water forms a hydrate . the term “ polymorph ” or “ polymorphism ” as used herein refers to a compound of this invention present in different crystal lattice forms . the term “ ester ” as used herein refers to a derivative of a compound of this invention derived from an oxoacid group and a hydroxyl group , either one of which can be present at the compound of this invention . the term “ tautomer ” as used herein refers to an isomer readily interconverted from a compound of this invention by e . g ., migration of a hydrogen atom or proton . the term “ pharmaceutically acceptable derivative or prodrug ” as used herein , refers to any pharmaceutically acceptable salt , ester , salt of an ester or other derivative of a compound of this invention , which , upon administration to a recipient , is capable of providing , either directly or indirectly , a compound of this invention or a pharmaceutically active metabolite or residue thereof . particularly favored derivatives or prodrugs are those that increase the bioavailability of the compounds of this invention when such compounds are administered to a patient ( e . g ., by allowing orally administered compound to be more readily absorbed into blood ) or which enhance delivery of the parent compound to a biological compartment ( e . g ., the brain or lymphatic system ). pharmaceutically acceptable prodrugs of the compounds described herein include , but are not limited to , esters , carbonates , thiocarbonates , n - acyl derivatives , n - acyloxyalkyl derivatives , quaternary derivatives of tertiary amines , n - mannich bases , schiff bases , amino acid conjugates , phosphate esters , metal salts and sulfonate esters . various forms of prodrugs are well known in the art . see for example design of prodrugs , bundgaard , a . ed ., elseview , 1985 and method in enzymology , widder , k . et al ., ed . ; academic , 1985 , vol . 42 , p . 309 - 396 ; bundgaard , h . “ design and application of prodrugs ” in a textbook of drug design and development , krosgaard - larsen and h . bundgaard , ed ., 1991 , chapter 5 , p . 113 - 191 ; and bundgaard , h ., advanced drug delivery review , 1992 , 8 , 1 - 38 , each of which is incorporated herein by reference . the prodrugs described herein include , but are not limited to , the following groups and combinations of these groups ; amine derived prodrugs : hydroxy prodrugs include , but are not limited to acyloxy - alkyl esters , alkoxycarbonyloxyalkyl esters , alkyl esters , aryl esters and disulfide containing esters . the terms “ enhance ” or “ enhancing ,” as used herein , means to increase or prolong either in potency or duration of a desired effect . thus , in regard to enhancing the effect of therapeutic agents , the term “ enhancing ” refers to the ability to increase or prolong , either in potency or duration , the effect of other therapeutic agents on a system . an “ enhancing - effective amount ,” as used herein , refers to an amount adequate to enhance the effect of another therapeutic agent in a desired system . the terms “ pharmaceutical combination ”, “ administering an additional therapy ”, “ administering an additional therapeutic agent ” and the like , as used herein , refer to a pharmaceutical therapy resulting from mixing or combining more than one active ingredient and includes both fixed and non - fixed combinations of the active ingredients . the term “ fixed combination ” means that at least one of the compounds described herein , and at least one co - agent , are both administered to a patient simultaneously in the form of a single entity or dosage . the term “ non - fixed combination ” means that at least one of the compounds described herein , and at least one co - agent , are administered to a patient as separate entities either simultaneously , concurrently or sequentially with variable intervening time limits , wherein such administration provides effective levels of the two or more compounds in the body of the patient . these also apply to cocktail therapies , e . g . the administration of three or more active ingredients . the terms “ co - administration ”, “ administered in combination with ” and their grammatical equivalents or the like , as used herein , are meant to encompass administration of the selected therapeutic agents to a single patient , and are intended to include treatment regimens in which the agents are administered by the same or different route of administration or at the same or different times . in some embodiments the compounds described herein will be co - administered with other agents . these terms encompass administration of two or more agents to an animal so that both agents and / or their metabolites are present in the animal at the same time . they include simultaneous administration in separate compositions , administration at different times in separate compositions , and / or administration in a composition in which both agents are present . thus , in some embodiments , the compounds of the invention and the other agent ( s ) are administered in a single composition . the term “ metabolite ,” as used herein , refers to a derivative of a compound which is formed when the compound is metabolized . the term “ active metabolite ,” as used herein , refers to a biologically active derivative of a compound that is formed when the compound is metabolized . the term “ metabolized ,” as used herein , refers to the sum of the processes ( including , but not limited to , hydrolysis reactions and reactions catalyzed by enzymes ) by which a particular substance is changed by an organism . thus , enzymes may produce specific structural alterations to a compound . for example , cytochrome p450 catalyzes a variety of oxidative and reductive reactions while uridine diphosphate glucuronyltransferases catalyze the transfer of an activated glucuronic - acid molecule to aromatic alcohols , aliphatic alcohols , carboxylic acids , amines and free sulphydryl groups . further information on metabolism may be obtained from the pharmacological basis of therapeutics , 9th edition , mcgraw - hill ( 1996 ). described herein are compounds of formula i , pharmaceutically acceptable salts , solvates , polymorphs , esters , tautomers or prodrugs thereof , x and w are independently selected from n , o , s or cr 2 ; r 1 is selected from the group consisting of h , halogen , c 1 - c 6 alkyl , c 3 - c 6 cycloalkyl , c 2 - c 6 alkenyl , c 5 - c 6 cycloalkenyl or c 2 - c 6 alkynyl ; wherein each alkyl , cycloalkyl , alkenyl , cycloalkenyl or alkynyl group is optionally substituted with 1 - 3 substituents selected independently from the group consisting of halogen , hydroxy , amino , alkylamino , dialkylamino , heterocyclyl , c 1 - c 4 alkyl , c 1 - c 4 alkoxy , cyano , cyanomethyl , trifluoromethyl , difluoromethoxy and phenyl , and one or two ring carbon atoms of said c 3 - c 6 cycloalkyl groups are optionally replaced with , independently , o , n , or s ; and r 2 is selected from the group consisting of h , halogen , c 1 - c 10 alkyl , c 1 - c 10 alkoxy , c 2 - c 10 alkenyl , c 2 - c 10 alkynyl , c 3 - c 10 cycloalkyl , c 3 - c 10 cycloalkoxy , c 3 - c 10 cycloalkylalkyl , aryl , arylalkyl , heteroaryl , heteroarylalkyl , heterocyclyl , and heterocyclylalkyl , where each alkyl , alkenyl , alkynyl , cycloalkyl , aryl , heteroaryl and heterocyclyl is unsubstituted or substituted with 1 - 3 substituents selected independently from halogen , hydroxyl , c 1 - c 4 alkyl , c 1 - c 4 alkoxy , cyano trifluoromethyl , difluoromethoxy , phenyl or substituted phenyl with 1 - 3 substituents selected independently from halogen , hydroxyl , c 1 - c 4 alkyl , c 1 - c 4 alkoxy , cyano trifluoromethyl , or difluoromethoxy ; r 3 is selected from the group consisting of h , c 1 - c 10 alkyl , c 1 - c 10 alkoxy , c 2 - c 10 alkenyl , c 2 - c 10 alkynyl , c 3 - c 10 cycloalkyl , c 3 - c 10 cycloalkoxy , c 3 - c 10 cycloalkylalkyl , aryl , arylalkyl , heteroaryl , heteroarylalkyl , heterocyclyl , and heterocyclylalkyl , where each alkyl , alkoxy , alkenyl , alkynyl , cycloalkyl , aryl , heteroaryl and heterocyclyl is unsubstituted or substituted with 1 - 3 substituents selected independently from halogen , hydroxyl , c 1 - c 4 alkyl , c 1 - c 4 alkoxy , cyano trifluoromethyl , difluoromethoxy , phenyl or substituted phenyl with 1 - 3 substituents selected independently from halogen , hydroxyl , c 1 - c 4 alkyl , c 1 - c 4 alkoxy , cyano trifluoromethyl , or difluoromethoxy ; r 3 ′ is selected from the groups consisting of h , c 1 - c 6 alkyl , c 2 - c 6 alkenyl and c 2 - c 6 alkynyl ; r 4 , r 5 , r 6 , r 7 and r 8 are independently selected from h , halogen , cyano , nitro , trifluoromethyl sr 9 , or 9 , c ( o ) r 9 , nr 10 c ( o ) or 12 , oc ( o ) r 9 , nr 10 s ( o ) j r 12 , s ( o ) j nr 9 r 10 , s ( o ) j nr 10 c ( o ) r 9 , c ( o ) nr 10 s ( o ) j r 12 , s ( o ) j r 12 , nr 10 ( o ) r 9 , c ( o ) nr 9 r 10 , nr 10 c ( o ) nr 9 r 10 , nr 11 c ( ncn ) nr 9 r 10 , nr 9 r 10 and c 1 - c 10 alkyl , c 2 - c 10 alkenyl , c 2 - c 10 alkynyl , c 3 - c 10 cycloalkyl , c 3 - c 10 cycloalkylalkyl , s ( o ) j ( c 1 - c 6 alkyl ), s ( o ) j ( cr 10 r 11 ) m - aryl , aryl , arylalkyl , heteroaryl , heteroarylalkyl , heterocyclyl , heterocyclylalkyl , o ( cr 9 r 10 ) m - aryl , n r11 ( cr 10 r 11 ) m - aryl , o ( cr 10 r 11 ) m - heteroaryl , nr 10 ( cr 10 r 11 ) m - heteroaryl , o ( cr 10 r 11 ) m - heterocyclyl , nr 10 ( cr 11 r 11 ) m - heterocyclyl , and s ( c 1 - c 2 alkyl ) optionally substituted with 1 - 5 fluorine atoms ; r 9 is selected from the group consisting of hydrogen , trifluoromethyl , c 1 - c 10 alkyl , c 2 - c 10 alkenyl , c 2 - c 10 alkynyl , c 3 - c 10 cycloalkyl , c 3 - c 10 cycloalkylalkyl , aryl , arylalkyl , heteroaryl , heteroarylalkyl , heterocyclyl , and heterocyclylalkyl , where each alkyl , alkenyl , alkynyl , cycloalkyl , aryl , heteroaryl and heterocyclyl is unsubstituted or substituted with 1 - 3 substituents independently selected from the group consisting of halogen , c 1 - c 4 alkyl , hydroxyl and amino ; r 10 is selected from hydrogen or c 1 - c 6 alkyl where alkyl may be unsubstituted or substituted with 1 - 3 substituents independently selected from the group consisting of halogen , c 1 - c 4 alkyl , hydroxyl and amino ; or r 9 and r 10 can be taken together with the atom to which they are attached to form a 4 to 10 membered heteroaryl or heterocyclic ring , each of which is unsubstituted or substituted with 1 - 3 substituents independently selected from the group consisting of halogen , c 1 - c 4 alkyl , hydroxyl and amino ; r 11 is selected from hydrogen or c 1 - c 6 alkyl where alkyl may be unsubstituted or substituted with 1 - 3 substituents independently selected from the group consisting of halogen , c 1 - c 4 alkyl , hydroxyl and amino ; or r 10 and r 11 can be taken together with the atom to which they are attached to form a 4 to 10 membered carbocyclic , heteroaryl or heterocyclic ring , each of which is unsubstituted or substituted with 1 - 3 substituents independently selected from the group consisting of halogen , c 1 - c 4 alkyl , hydroxyl and amino ; r 12 is selected from trifluoromethyl , c 1 - c 10 alkyl , c 3 - c 10 cycloalkyl , aryl , arylalkyl , heteroaryl , heteroarylalkyl , heterocyclyl , and heterocyclylalkyl , where each alkyl , cycloalkyl , aryl , heteroaryl and heterocyclyl unsubstituted or substituted with 1 - 3 substituents independently selected from the group consisting of halogen , c 1 - c 4 alkyl , hydroxyl and amino ; r 13 is selected from the groups consisting of h , c 1 - c 6 alkyl , c 2 - c 6 alkenyl and c 2 - c 6 alkynyl ; r 14 is selected from the groups consisting of h , c 1 - c 6 alkyl , c 2 - c 6 alkenyl and c 2 - c 6 alkynyl ; methods for synthesizing the compounds described herein are provided . in some embodiments , the compounds described herein can be prepared by the methods described below . the procedures and examples below are intended to illustrate those methods . neither the procedures nor the examples should be construed as limiting the invention in any way . compounds described herein may also be synthesized using standard synthetic techniques known to those of skill in the art or using methods known in the art in combination with methods described herein . the compounds of formula i wherein r 3 ′ is hydrogen , r 13 is hydrogen , and r 14 is methyl shown as the following formula is taken as an example to illustrate the preparation of the compounds of formula i . scheme 1 above illustrates the preparation of 5 - membered ring [ b ] pyridone amide of ( 6 ). deporton of 5 - membered ring di - ester ( 1 ) with base as nah , lihmds , etc . followed by treatment with carbodiimide ( 2 ) afforded pyridone ester ( 3 ). coupling the amines ( 5 ) by treatment the pyridone ester ( 3 ) with lihmds or by coupled with pyridone acid ( 4 ) after hydrolysis from pyridone ester ( 3 ), gave desired pyridone amides ( 6 ). procedure a : to the solution of aniline ( 5 g , 1 eq .) in 50 ml etoh was added methylisothiocyanate ( 1 . 4 eq .) at room temperature , the mixture was then heated under reflux overnight . after cooling down , the precipitate was collected and washed with 50 ml petroleum ether / dichloromethane ( 10 / 1 ) to afford the thiourea . to a solution of 10 mmol of the thiourea , 3 . 0 g ( 30 mmol ) of triethylamine , and 50 mg of 4 - dimethylamino - pyridine ( dmap ) in 100 ml of methylene chloride is added dropwise 2 . 3 g ( 20 mmol ) of methanesulfonyl chloride ( on a larger scale some cooling may be required to keep the reaction near room temperature ). after addition is complete , the resulting yellow solution is stirred at room temperature for 5 minutes . approximately two - thirds of the solvent is removed under reduced pressure and the remaining mixture is filtered through a pad of silica gel using methylene chloride to elute the product . a second pad filtration is needed to remove any final traces of polar material to afford the cabodiimide ( ref . : syn . commun ., 1995 , 25 , 43 - 47 .) thiourea : 1 h nmr ( 400 mhz , dmso - d6 ) δ 9 . 30 ( s , 1h ), 7 . 86 ( s , 1h ), 7 . 67 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 52 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 7 . 44 ( t , j = 8 . 4 hz , 1h ), 2 . 89 ( d , j = 4 . 0 hz , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 40 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 35 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 6 . 79 ( t , j = 8 . 4 hz , 1h ), 3 . 18 ( s , 3h ). thiourea : 1 h nmr ( 400 mhz , dmso - d6 ) δ 9 . 23 ( s , 1h ), 7 . 88 ( s , 1h ), 7 . 62 - 7 . 66 ( m , 2h ), 7 . 40 ( t , j = 8 . 8 hz , 1h ), 2 . 91 ( d , j = 4 . 4 hz , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 24 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 17 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 6 . 93 ( t , j = 8 . 4 hz , 1h ), 3 . 17 ( s , 3h ). thiourea : 1 h nmr ( 400 mhz , dmso - d6 ) δ 9 . 22 ( s , 1h ), 7 . 90 ( s , 1h ), 7 . 76 ( s , 1h ), 7 . 57 - 7 . 59 ( m , 1h ), 7 . 50 - 7 . 53 ( m , 1h ), 2 . 90 ( d , j = 4 . 4 hz , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 47 ( d , j = 2 . 0 hz , 1h ), 7 . 25 ( dd , j = 2 . 0 & amp ; 8 . 8 hz , 1h ), 6 . 95 ( d , j = 8 . 8 hz , 1h ), 3 . 16 ( s , 3h ). thiourea : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 83 ( d , j = 1 . 6 hz , 1h ), 7 . 64 ( dd , j = 1 . 6 & amp ; 8 . 4 hz , 1h ), 7 . 50 ( s , 1h ), 7 . 27 ( s , br , 1h ), 6 . 04 ( s , 1h ), 3 . 15 ( d , j = 4 . 8 hz , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 67 ( d , j = 2 . 0 hz , 1h ), 7 . 46 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 6 . 84 ( d , j = 8 . 4 hz , 1h ), 3 . 18 ( s , 3h ). thiourea : 1 h nmr ( 400 mhz , dmso - d6 ) δ 9 . 22 ( s , 1h ), 7 . 11 ( s , 1h ), 7 . 63 - 7 . 67 ( m , 2h ), 7 . 39 ( dd , j = 2 . 4 & amp ; 8 . 8 hz , 1h ), 2 . 92 ( d , j = 4 . 4 hz , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 37 ( d , j = 2 . 0 hz , 1h ), 7 . 15 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 7 . 04 ( d , j = 8 . 4 hz , 1h ), 3 . 18 ( s , 3h ). thiourea : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 52 ( s , 1h ), 7 . 27 ( s , br , 1h ), 7 . 04 ( d , j = 8 . 8 hz , 2h ), 5 . 98 ( s , br , 1h ), 3 . 14 ( d , j = 4 . 4 hz , 3h ), 2 . 49 ( s , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 6 . 91 - 6 . 99 ( m , 3h ), 3 . 16 ( s , 3h ), 2 . 46 ( s , 3h ). thiourea : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 53 ( s , 1h ), 7 . 30 - 7 . 33 ( m , 2h ), 7 . 17 ( dd , j = 2 . 4 & amp ; 8 . 4 hz , 1h ), 5 . 94 ( s , br , 1h ), 3 . 14 ( d , j = 4 . 8 hz , 3h ), 2 . 50 ( s , 3h ); carbodiimide : 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 24 ( d , j = 2 . 4 hz , 1h ), 7 . 01 - 7 . 08 ( m , 2h ), 3 . 16 ( s , 3h ), 2 . 46 ( s , 3h ). procedure b : to a stirred solution of diester ( 1 . 0 eq .) in anhydrous thf was added nah ( 1 . 1 eq ., 60 %) portionwise at 0 ° c . then carbodiimide ( 1 . 4 eq ., prepared from procedure a ) was added slowly with dropping funnel within 1 h , the mixture was then stirred at room temperature overnight . after the addition of water and ethyl acetate , the freshly formed white precipitate was collected and washed with ethyl acetate , dried with infrared lamp to get the desired product . in some cases , the product did not precipitate , then it need purification by column chromatography . procedure c : to a stirred solution of the acid ( 1 . 0 eq . ), hobt ( 1 . 5 eq .) and edci ( 1 . 5 eq .) in dry dmf , was added hydroxylamine ( 1 . 1 eq .) and et 3 n ( 1 . 5 eq .). the mixture was then stirred at room temperature overnight . then water was added , extracted with ethyl acetate , dried ( mgso 4 ) and concentrated in vacuum , the residue was purified by column chromatography on silica gel to get the desired product . to a stirred solution of the dimethyl 3 - oxopentanedioate ( 50 g , 287 mmol ) in anhydrous pyridine ( 100 ml ), was added the 2 - chloroacetaldehyde ( 67 . 6 g , 345 mmol , 1 . 2 eq ., 40 %) under the ice - bath . the mixture was stirred under 50 ° c . overnight . after removing the pyridine the under reduced pressure , ethyl acetate was added , and the organic phase was washed with water , dried ( na 2 so 4 ) and concentrated under the reduced pressure . the residue was purified by flash column chromatography on silica gel to get the desired product ( yield = 61 %). 1 h nmr ( 400 mhz , cdcl 3 ) 7 . 34 ( d , j = 2 . 0 hz , 1h ), 6 . 70 ( d , j = 2 . 0 hz , 1h ), 4 . 09 ( s , 2h ), 3 . 83 ( s , 3h ), 3 . 73 ( s , 3h ). according to the procedure b : to a stirred solution of furan diester ( 4 . 8 g , 1 . 0 eq .) in anhydrous thf ( 50 ml ) was added nah ( 1 . 1 g , 1 . 1 eq ., 60 %) portionwise at 0 ° c . then 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene aniline ( 7 . 35 g , 1 . 1 eq ., prepared from procedure a ) was added slowly with dropping funnel within 1 h and the mixture was then stirred at room temperature overnight . after the addition of water and ethyl acetate , the freshly formed white precipitate was collected and washed with ethyl acetate , dried with infrared lamp to get the desired product ( yield = 59 %). 1 h nmr ( 400 mhz , dmso - d6 ) δ 9 . 16 ( s , 1h ), 7 . 91 ( d , j = 2 . 0 hz , 1h ), 7 . 65 - 7 . 68 ( dd , j = 1 . 8 & amp ; 10 . 6 hz , 1h ), 7 . 37 - 7 . 39 ( dd , j = 1 . 0 & amp ; 8 . 2 hz , 1h ), 6 . 98 - 6 . 99 ( d , j = 2 . 0 hz , 1h ), 6 . 64 - 6 . 68 ( t , j = 8 . 8 hz , 1h ), 3 . 69 ( s , 3h ), 3 . 30 ( s , 3h ). to a stirred solution ( thf : meoh : h 2 o = 1 : 1 : 1 ) of the methyl ester , was added k 2 co 3 ( 1 . 2 eq . ), the reaction mixture was then stirred at 50 ° c . for 3 h . then water was added and the mixture was washed with ethyl acetate twice , water layer separated , acidified with 2 n hcl , the freshly formed white precipitate was collected , washed with water , dried over infrared lamp to get the desired product ( 62 %). 1 h nmr ( 400 mhz , dmso - d6 ) δ 7 . 82 ( d , j = 2 . 0 hz , 1h ), 7 . 65 ( dd , j = 2 . 0 & amp ; 10 . 8 hz , 1h ), 7 . 38 ( d , j = 8 . 4 hz , 1h ), 6 . 90 ( d , j = 2 . 0 hz , 1h ), 6 . 57 ( t , j = 8 . 4 hz , 1h ), 3 . 20 ( s , 3h ). to a stirred solution of the acid ( 4 g , 1 . 0 eq . ), hobt ( 1 . 9 g , 1 . 5 eq .) and edci ( 2 . 7 g , 1 . 5 eq .) in dry dmf ( 25 ml ), was added o -( 2 -( vinyloxy ) ethyl ) hydroxylamine ( 1 . 1 g , 1 . 1 eq .) and et 3 n ( 1 . 4 g , 1 . 5 eq ., 2 . 2 ml ). the mixture was then stirred at room temperature overnight . then added with water , extracted with ethyl acetate , dried ( mgso 4 ) and concentrated in vacuum . the residue was then dissolved in meoh ( 50 ml ) and 2n hcl ( 10 ml ) was added at room temperature . after stirring at r . t . for 0 . 5 h , the mixture was concentrated in vacuum , then water and ethyl acetate was added , water layer was extracted with etoac , washed with water , dried over mgso 4 and concentrated under reduced pressure . the residue was purified by flash column chromatography on silica gel to obtain the desired product ( 53 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 82 ( s , 1h ), 9 . 84 ( s , 1h ), 7 . 58 - 7 . 48 ( m , 2h ), 7 . 47 - 7 . 28 ( m , 1h ), 7 . 02 ( s , 1h ), 6 . 56 ( t , j = 8 . 8 hz , 1h ), 4 . 12 - 4 . 06 ( m , 3h ), 3 . 79 - 3 . 76 ( m , 2h ), 3 . 31 ( s , 3h ); m / z = 411 [ m - nhoch 2 ch 2 oh ] + , 488 [ m + 1 ] + . ( d , l )- proline ( 1 . 9 g , 17 mmol ) was added to a stirred ice - cooled solution of propionaldehyde ( 5 . 0 g , 86 mmol ) in 50 ml chcl 3 followed by the addition of n - chlorosuccinimide ( 12 . 6 g , 92 mmol ). the reaction mixture was stirred after 1 h then allowed to warm to ambient temperature and stirred until the aldehydes was completely consumed . 7 hours later , pentane was added to the reaction mixture and the precipitated n - chlorosuccinimide succinimide and the catalysts were filtered off . the solution was washed with water 2 times and dried with na 2 so 4 , the solution was directly used to next step . to a solution of dimethyl 3 - oxopentanedioate ( 13 . 5 g , 77 mmol ) in 20 ml pyridine , 2 - chloropropanal in pentane ( 77 mmol ) was added dropwise at room temperature , after the addition , the reaction mixture was heat at 60 ° c . overnight , then water and etoac were added , the mixture was extracted with etoac thrice , the combined organic layers was dried over na 2 so 4 , filtered , the filtrate was concentrated in vacuum , the residue was purified by column chromatography on silica gel to get the desired product ( 11 . 7 g , 70 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 6 . 27 ( s , 1h ), 4 . 03 ( s , 2h ), 0 . 79 ( s , 3h ), 3 . 73 ( s , 3h ), 2 . 28 ( s , 3h ); m / z = 213 [ m + 1 ] + . according to the procedure b , methyl 2 -( 2 - methoxy - 2 - oxoethyl )- 5 - methylfuran - 3 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 67 ( s , 1h ), 7 . 47 - 7 . 50 ( dd , j = 1 . 6 & amp ; 9 . 6 hz , 1h ), 7 . 34 - 7 . 36 ( dd , j = 1 . 2 & amp ; 8 . 4 hz , 1h ), 6 . 54 ( s , 1h ), 6 . 38 - 6 . 42 ( t , j = 8 . 4 hz , 1h ), 3 . 95 ( s , 3h ), 3 . 36 ( s , 3h ), 2 . 45 ( m , 3h ); m / z = 457 [ m + 1 ] + . following the same procedure as step c , example 1 described , the title product was obtained via column chromatography purification . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 67 ( s , 1h ), 9 . 87 ( s , 1h ), 7 . 48 ( d , j = 10 . 0 hz , 1h ), 7 . 37 ( d , j = 8 . 0 hz , 1h ), 6 . 58 ( s , 1h ), 6 . 50 ( t , j = 8 . 0 hz , 1h ), 4 . 10 - 4 . 11 ( m , 3h ), 3 . 78 ( m , 2h ), 3 . 30 ( s , 3h ), 2 . 48 ( s , 3h ); m / z = 502 [ m + 1 ] + , 425 [ m - nhoch 2 ch 2 oh ] + . following the same procedure as example 1 described , using 4 - bromo - 2 - fluoro - n -(( methylimino ) methylene ) aniline as starting material to get the product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 84 ( s , 1h ), 9 . 87 ( s , 1h ), 7 . 52 ( d , j = 2 . 0 hz , 1h ), 7 . 33 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 22 ( d , j = 8 . 4 hz , 1h ), 7 . 02 ( d , j = 2 . 0 hz , 1h ), 6 . 70 ( t , j = 8 . 4 hz , 1h ), 4 . 09 - 4 . 12 ( m , 3h ), 3 . 77 ( m , 2h ), 3 . 30 ( s , 3h ); m / z = 363 [ m - nhoch 2 ch 2 oh ] + , 440 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was coupling with ( s )— o -( 2 -( tert - butyl dimethylsilyloxy ) propyl ) hydroxylamine ( made according to wo2010003025 a1 ), followed by de - tbs by 2 n hcl in meoh to afford the title compound . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 86 ( s , 1h ), 9 . 91 ( s , 1h ), 7 . 52 ( d , j = 1 . 6 hz , 1h ), 7 . 48 ( d , j = 8 . 4 hz , 1h ), 7 . 39 ( d , j = 8 . 4 hz , 1h ), 7 . 01 ( d , j = 1 . 6 hz , 1h ), 6 . 55 ( t , j = 8 . 4 hz , 1h ), 4 . 41 ( s , 1h ), 4 . 01 - 3 . 98 ( m , 2h ), 3 . 77 - 3 . 71 ( m , 1h ), 3 . 30 ( s , 3h ); m / z = 411 [ m - nhoch 2 ch ( ch 3 ) oh ] + , 502 [ m + 1 ] + . to a solution of ( r )- propane - 1 , 2 - diol ( 20 g , 0 . 36 mmol ) and triethylamine ( 50 . 8 ml , 0 . 34 mmol ) in ch 2 cl 2 was added tert - butylchlorodimethylsilane ( 39 . 6 g , 0 . 26 mmol ). after stirring overnight at room temperature , the reaction mixture was washed one time each with 1 n aqueous hcl solution , water , and a 1 : 1 saturated solution of nahco 3 and brine . the organic layer was dried over na 2 so 4 , then filtered and concentrated . the crude title compound was used without further purification in the next step . dead ( 52 . 7 g ) was added dropwise to a solution of ( r )- 1 -( tert - butyldimethylsilyloxy ) propan - 2 - ol ( 44 . 3 g ), pph 3 ( 61 g ), and n - hydroxyphthalimide ( 38 g ) in thf ( 500 ml ) at 0 ° c . after stirring for 10 min at 0 ° c ., the reaction mixture was brought to room temperature and stirring was continued for a further 24 h . the reaction mixture was filtered through a coarse glass funnel and concentrated in vacuo . the residue was purified by silica chromatography to afford the title compound ( 97 %) as a clear oil . n - methylhydrazine ( 13 . 12 g , 0 . 85 mmol ) was added to a solution of ( s )- 2 -( 1 -( tert - butyldimethylsilyloxy ) propan - 2 - yloxy ) isoindoline - 1 , 3 - dione ( 91 g , 0 . 27 mmol ) in ch 2 cl 2 ( 300 ml ). after stirring for 1 h at room temperature , the white precipitate was filtered off and the reaction mixture was concentrated in vacuo to afford the title compound ( 37 g , 66 %) as a yellow oil . 1 h nmr ( 400 mhz , cdcl 3 ) δ 5 . 30 ( s , 2h ), 3 . 51 - 3 . 67 ( m , 3h ), 1 . 03 - 1 . 06 ( m , 3h ), 0 . 79 - 0 . 83 ( m , 9h ), 0 . 00 ( s , 6h ). according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was coupling with ( s )— o -( 1 -( tert - butyl dimethylsilyloxy ) propan - 2 - yl ) hydroxylamine to get the title compound . to a stirred solution of ( s )— n -( 1 -( tert - butyldimethylsilyloxy ) propan - 2 - yloxy )- 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxamide ( 100 mg ) in meoh ( 10 ml ) was added 2 n hcl ( aq .) ( 3 ml ) at room temperature and the mixture was stirred for 10 min . then concentrated in vacuum , residue was partitioned between water and ethyl acetate , water phase was extracted thrice and the combined organic phase was washed with water , dried ( mgso 4 ) and concentrated under reduced pressure . the residue was purified by flash column chromatography on silica gel to obtain the title product ( 53 mg ). 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 83 ( s , 1h ), 9 . 75 ( s , 1h ), 7 . 53 ( d , j = 1 . 6 hz , 1h ), 7 . 47 ( d , j = 8 . 4 hz , 1h ), 7 . 40 ( d , j = 8 . 4 hz , 1h ), 7 . 02 ( d , j = 1 . 6 hz , 1h ), 6 . 55 ( t , j = 8 . 4 hz , 1h ), 4 . 20 - 4 . 11 ( m , 2h ), 3 . 71 - 3 . 69 ( m , 1h ), 3 . 52 - 3 . 48 ( m , 1h ), 3 . 30 ( s , 3h ), 1 . 34 ( d , j = 6 . 4 hz , 3h ); m / z = 411 [ m - nhoch ( ch 3 ) ch 2 oh ] + , 502 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with o -( 2 - methoxyethyl ) hydroxylamine ( made according to wo2008070758 a1 ) to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 98 ( s , 1h ), 10 . 17 ( s , 1h ), 7 . 46 - 7 . 50 ( m , 2h ), 7 . 36 - 7 . 38 ( d , j = 8 . 4 hz , 1h ), 7 . 01 - 7 . 02 ( dd , j = 0 . 8 hz & amp ; 2 . 0 hz , 1h ), 6 . 47 - 6 . 52 ( t , j = 8 . 4 hz , 1h ), 4 . 19 - 4 . 22 ( m , 2h ), 3 . 70 - 3 . 72 ( m , 2h ), 3 . 45 ( s , 3h ), 3 . 32 ( s , 3h ); j = 8 . 4 hz , 1h ), 7 . 02 ( d , j = 1 . 6 hz , 1h ), 6 . 55 ( t , j = 8 . 4 hz , 1h ), 4 . 20 - 4 . 11 ( m , 2h ), 3 . 71 - 3 . 69 ( m , 1h ), 3 . 52 - 3 . 48 ( m , 1h ), 3 . 30 ( s , 3h ), 1 . 34 ( d , j = 6 . 4 hz , 3h ); m / z = 411 [ m - nhoch 2 ch 2 och 3 ] + , 502 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with o -( cyclopropylmethyl ) hydroxylamine ( made according to wo2005054179 ) to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 99 ( s , 1h ), 9 . 86 ( s , 1h ), 7 . 46 - 7 . 52 ( m , 2h ), 7 . 36 - 7 . 38 ( d , j = 8 . 4 hz , 1h ), 7 . 26 - 7 . 27 ( d , j = 0 . 4 hz , 1h ) 7 . 01 - 7 . 02 ( t , j = 0 . 8 hz , 1h ), 6 . 48 - 6 . 52 ( t , j = 8 . 4 hz , 1h ), 3 . 86 - 3 . 88 ( d , j = 7 . 2 hz , 2h ), 3 . 32 ( s , 3h ), 1 . 19 - 1 . 26 ( m , 1h ), 0 . 61 - 0 . 66 ( m , 2h ) 0 . 33 - 0 . 37 ( m , 2h ); m / z = 498 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with 2 -( aminooxy )- 2 - methylpropan - 1 - ol hydrochloride ( made according to wo2010003025 a1 ) to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 80 ( s , 1h ), 9 . 46 ( s , 1h ), 7 . 47 - 7 . 53 ( m , 2h ), 7 . 38 - 7 . 41 ( dd , j = 0 . 8 hz & amp ; 8 . 4 hz , 1h ), 7 . 02 - 7 . 03 ( d , j = 2 . 0 hz , 1h ), 6 . 53 - 6 . 57 ( t , j = 8 . 4 hz , 1h ), 4 . 50 - 4 . 52 ( t , j = 7 . 2 hz , 1h ), 3 . 39 - 3 . 41 ( d , j = 7 . 2 hz , 2h ), 3 . 31 ( s , 3h ), 1 . 24 - 1 . 34 ( m , 6h ); m / z = 411 [ m - nhoc ( ch 3 ) 2 ch 2 oh ] + , 516 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with ( r )— o -(( 2 , 2 - dimethyl - 1 , 3 - dioxolan - 4 - yl ) methyl ) hydroxylamine ( made according to tetrahedron letters , 2006 , 47 , 7607 - 7609 ), followed by treating with 2 n hcl in meoh to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 79 ( s , 1h ), 9 . 99 ( s , 1h ), 7 . 48 - 7 . 53 ( m , 2h ), 7 . 39 - 7 . 41 ( d , j = 8 . 4 hz , 1h ), 7 . 01 - 7 . 02 ( d , j = 2 . 0 hz , 1h ), 6 . 54 - 6 . 58 ( t , j = 8 . 4 hz , 1h ), 4 . 44 - 4 . 45 ( d , j = 3 . 2 hz , 1h ), 4 . 03 - 4 . 14 ( m , 2h ), 3 . 97 - 3 . 99 ( m , 1h ), 3 . 63 - 3 . 76 ( m , 2h ), 3 . 30 ( s , 3h ), 2 . 38 - 2 . 41 ( t , 1h ); m / z = 411 [ m - nhoch 2 ch ( oh ) ch 2 oh ] + , 518 [ m + 1 ] + . to a cucn ( 109 mg , 4 %) was added dry thf ( 100 ml ) under n 2 , and the suspension was then cooled to − 15 ° c . memgbr ( 3 m in ether , 13 ml ) was added dropwise and the reaction mixture was stirred at − 15 ° c . for 15 min , then , ( s )- benzyl glycidyl ether ( 5 g , 30 mmol ) was added and the mixture was stirred at the same temperature for 1 h . the reaction mixture was quenched with aqueous sat . nh 4 cl and 25 % nh 3 solution and the resulting mixture was stirred at room temperature until the color turned blue . the organic layers were extracted with ether washed with brine , dried by mgso 4 , filtered and evaporated in vacuo . the material was used without further purification in the subsequent step . 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 29 - 7 . 36 ( m , 5h ), 4 . 54 ( s , 2h ), 3 . 72 - 3 . 74 ( m , 1h ), 3 . 50 ( dd , j = 2 . 8 hz & amp ; 9 . 6 hz , 1h ), 3 . 33 ( dd , j = 8 . 0 hz & amp ; 9 . 6 hz , 1h ), 2 . 52 ( d , j = 3 . 2 hz , 1h ), 1 . 44 - 1 . 49 ( m , 2h ), 0 . 95 ( t , j = 7 . 6 hz , 3h ). to a solution of ( s )- 1 -( benzyloxy ) butan - 2 - ol ( 41 . 4 g , 0 . 23 mol ) in ch 2 cl 2 ( 200 ml ) was added imidazole ( 23 . 5 g , 0 . 36 mol ) and tbscl ( 36 . 4 g , 0 . 24 mol ) and dmap ( 600 mg , 0 . 005 mol ). the reaction mixture was stirred at room temperature overnight . the reaction was diluted with water and extracted with ch 2 cl 2 thrice , the combined organic extracts washed with brine and dried over mgso 4 , filtered , concentrated in vacuo to give the title product as a colorless oil ( 99 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 19 - 7 . 22 ( m , 5h ), 4 . 48 ( s , 2h ), 3 . 70 - 3 . 73 ( m , 1h ), 3 . 29 - 3 . 37 ( m , 2h ), 1 . 52 - 1 . 55 ( m , 1h ), 1 . 39 - 1 . 44 ( m , 1h ), 0 . 81 - 0 . 87 ( m , 12h ), 0 . 01 ( s , 6h ). to a solution of ( s )-( 1 -( benzyloxy ) butan - 2 - yloxy )( tert - butyl ) dimethylsilane in ethyl acetate ( 400 ml ) was added 20 % pd / c ( 2 g ). the reaction mixture was evacuated and flushed with h 2 , then stirred under an atmosphere of h 2 ( 1 atm ) overweekends . the reaction mixture was then filtered through celite and concentrated to afford the title compound ( 45 g , 97 % yield ) as clear oil , which was used without further purification in the next step . following the same procedure as step b , example 5 described , the title product was obtained . following the same procedure as step c , example 5 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ): δ 5 . 36 ( s , 2h ), 3 . 70 - 3 . 75 ( m , 1h ), 3 . 49 - 3 . 57 ( m , 2h ), 1 . 33 - 1 . 51 ( m , 2h ), 0 . 81 - 0 . 88 ( m , 12h ), 0 . 00 ( s , 6h ). according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was coupling with ( s )— o -( 2 -( tert - butyldimethyl silyloxy ) butyl ) hydroxylamine to give the title compound . following the same procedure as step e , example 5 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ): δ 10 . 87 ( s , 1h ), 9 . 92 ( s , 1h ), 7 . 53 ( d , j = 1 . 6 hz , 1h ), 7 . 51 ( d , j = 8 . 4 hz , 1h ), 7 . 47 ( d , j = 8 . 4 hz , 1h ), 7 . 02 ( d , j = 1 . 6 hz , 1h ), 6 . 56 ( t , j = 8 . 4 hz , 1h ), 4 . 33 ( s , 1h ), 4 . 06 - 4 . 04 ( m , 1h ), 3 . 82 - 3 . 80 ( m , 2h ), 3 . 30 ( s , 3h ), 1 . 58 - 1 . 41 ( m , 2h ), 0 . 97 ( t , j = 2 . 4 hz , 3h ); m / z = 411 [ m - nhoch 2 ch ( oh ) ch 2 ch 3 ] + , 516 [ m + 1 ] + . following the same procedure as example 1 described , using 2 , 4 - dichloro - n -(( methylimino ) methylene ) aniline as starting material to get the product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 80 ( s , 1h ), 9 . 86 ( s , 1h ), 7 . 54 ( d , j = 2 . 4 hz , 1h ), 7 . 49 ( d , j = 2 . 4 hz , 1h ), 7 . 15 ( dd , j = 2 . 4 & amp ; 8 . 4 hz , 1h ), 7 . 03 ( d , j = 2 . 4 hz , 1h ), 6 . 60 ( d , j = 8 . 8 hz , 1h ), 4 . 10 - 4 . 12 ( m , 2h ), 4 . 03 ( t , j = 6 . 4 hz , 1h ), 3 . 77 - 3 . 79 ( m , 2h ), 3 . 26 ( s , 3h ); m / z = 434 [ m + na ] + , 335 [ m - nhoch 2 ch 2 oh ] + , 412 [ m + 1 ] + . following the same procedure as example 1 described , using 4 - bromo - 2 - chloro - n -(( methylimino ) methylene ) aniline as starting material to get the product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 78 ( s , 1h ), 9 . 85 ( s , 1h ), 7 . 63 ( d , j = 2 . 4 hz , 1h ), 7 . 54 ( d , j = 2 . 0 hz , 1h ), 7 . 29 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 7 . 03 ( d , j = 2 . 4 hz , 1h ), 6 . 53 ( d , j = 8 . 4 hz , 1h ), 4 . 10 - 4 . 13 ( m , 2h ), 4 . 02 ( t , j = 6 . 4 hz , 1h ), 3 . 75 - 3 . 79 ( m , 2h ), 3 . 27 ( s , 3h ); m / z = 478 [ m + na ] + , 379 [ m - nhoch 2 ch 2 oh ] + , 456 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with o - methylhydroxylamine to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 99 ( s , 1h ), 9 . 86 ( s , 1h ), 7 . 52 ( d , j = 2 . 0 hz , 1h ), 7 . 48 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 38 ( d , j = 8 . 4 hz , 1h ), 7 . 02 ( d , j = 2 . 0 hz , 1h ), 6 . 51 ( t , j = 8 . 4 hz , 1h ), 3 . 91 ( s , 3h ), 3 . 33 ( s , 3h ); m / z = 458 [ m + 1 ] + , 426 [ m - och 3 ] + , 411 [ m - nhoch 3 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with o - ethylhydroxylamine to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 01 ( s , 1h ), 9 . 80 ( s , 1h ), 7 . 52 ( d , j = 2 . 0 hz , 1h ), 7 . 46 - 7 . 49 ( dd , j = 2 . 0 hz & amp ; 10 hz , 1h ), 7 . 36 - 7 . 38 ( d , j = 8 . 8 hz , 1h ), 7 . 01 - 7 . 02 ( d , j = 2 . 0 hz , 1h ), 6 . 48 - 6 . 52 ( t , j = 8 . 4 hz , 1h ), 4 . 08 - 4 . 13 ( m , 2h ), 3 . 33 ( s , 3h ), 1 . 33 - 1 . 37 ( m , 3h ); m / z = 472 [ m + 1 ] + , 426 [ m - och 2 ch 3 ] + , 411 [ m - nhoch 2 ch 3 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with ammonium hydroxide ( 25 %) to get the desired product . 1 h nmr ( 400 mhz , dmso - d6 ) δ 10 . 93 ( s , 1h ), 8 . 25 ( s , 1h ), 7 . 88 ( s , 1h ), 7 . 83 - 7 . 84 ( d , j = 3 hz , 1h ), 7 . 60 - 7 . 62 ( dd , j = 2 . 0 hz & amp ; 10 . 4 hz , 1h ), 7 . 51 ( s , 1h ), 7 . 40 - 7 . 42 ( d , j = 8 . 4 hz , 1h ), 6 . 62 - 6 . 66 ( t , j = 8 . 6 hz , 1h ), 4 . 08 - 4 . 13 ( m , 2h ), 3 . 33 ( s , 3h ); m / z = 428 [ m + 1 ] + , 411 [ m - nh 2 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with (±)- 3 - amino - 1 , 2 - propanediol to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 14 ( s , 1h ), 7 . 80 - 7 . 83 ( t , j = 5 . 4 hz , 1h ), 7 . 52 ( d , 1h ), 7 . 47 - 7 . 49 ( dd , j = 1 . 6 hz & amp ; 8 . 8 hz , 1h ), 7 . 36 - 7 . 38 ( d , j = 8 . 4 hz , 1h ), 6 . 47 - 6 . 51 ( t , j = 8 . 4 hz , 1h ), 3 . 89 - 3 . 92 ( m , 2h ), 3 . 56 - 3 . 69 ( m , 4h ), 3 . 32 ( s , 3h ), 2 . 84 - 2 . 86 ( d , j = 3 hz , 1h ), 2 . 57 - 2 . 60 ( t , 1h ); m / z = 502 [ m + 1 ] + , 411 [ m - nhch 2 ch ( oh ) ch 2 oh ] + . following the same procedure as example 1 described , using 2 - chloro - 4 - iodo - n -(( methylimino ) methylene ) aniline as starting material to get the product . 1 h nmr ( 400 mhz , dmso - d6 ) δ11 . 35 ( s , 1h ), 8 . 90 ( s , 1h ), 7 . 93 - 7 . 94 ( d , j = 2 . 4 hz , 1h ), 7 . 76 - 7 . 75 ( d , j = 2 . 0 hz , 1h ), 7 . 44 - 7 . 47 ( dd , j = 1 . 8 hz & amp ; 8 . 6 hz , 1h ), 7 . 01 - 7 . 02 ( d , j = 2 . 4 hz , 1h ), 6 . 44 - 6 . 46 ( d , j = 8 . 4 hz , 1h ), 4 . 65 ( s , 1h ), 3 . 69 - 3 . 71 ( t , j = 5 . 0 hz , 2h ), 3 . 48 - 3 . 49 ( d , j = 4 . 8 hz , 2h ), 3 . 31 ( s , 3h ); m / z = 427 [ m - nhoch 2 ch 2 oh ] + , 504 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with 3 - methoxypropan - 1 - amine then treated with bbr 3 at 0 ° c . to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 25 ( s , 1h ), 7 . 70 - 7 . 72 ( d , 1h ), 7 . 48 - 7 . 51 ( dd , j = 2 . 0 hz & amp ; 9 . 6 hz , 1h ), 7 . 35 - 7 . 37 ( d , j = 8 . 4 hz , 1h ), 7 . 26 ( s , 1h ), 7 . 02 ( d , 1h ), 6 . 46 - 6 . 51 ( t , j = 8 . 8 hz , 1h ), 3 . 68 - 3 . 72 ( m , 2h ), 3 . 61 - 3 . 66 ( m , 2h ), 3 . 33 ( s , 3h ), 1 . 57 - 1 . 86 ( m , 2h ); m / z = 486 [ m + 1 ] + , 411 [ m - nhch 2 ch 2 ch 2 oh ] + . according to wo 2005087779 , the title compound was made . 1 h nmr ( 400 mhz , dmso - d6 ) δ 7 . 46 ( d , j = 5 . 6 hz , 1h ), 7 . 36 ( d , j = 5 . 6 hz , 1h ), 4 . 21 ( s , 2h ), 3 . 73 ( s , 3h ), 3 . 61 ( s , 3h ). according to the procedure b , methyl 2 -( 2 - methoxy - 2 - oxoethyl ) thiophene - 3 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product . 1 h nmr ( 400 mhz , dmso - d6 ) δ 9 . 05 ( s , 1h ), 7 . 67 ( dd , j = 2 . 0 & amp ; 11 . 2 hz , 1h ), 7 . 45 - 7 . 57 ( m , 2h ), 7 . 38 ( d , j = 8 . 0 hz , 1h ), 6 . 65 ( t , j = 8 . 8 hz , 1h ), 3 . 69 ( s , 3h ), 3 . 35 ( s , 3h ). to a solution of methyl 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrothieno [ 3 , 2 - c ] pyridine - 7 - carboxylate ( 119 mg , 0 . 26 mmol ) in 5 ml thf was added o -( 2 -( vinyloxy ) ethyl ) hydroxylamine ( 33 mg , 0 . 32 mmol ). the solution was cooled to 0 ° c . and 1 ml 1 m lithium bis ( trimethylsilyl ) amide ( 1 mmol ) was added dropwise . the reaction mixture was then warmed to room temperature and stirred for 30 min , then quenched with nh 4 cl ( aq . ), and partitioned between ch 2 cl 2 and saturated nacl . the organic layers was separated , dried , filtered , purified by column chromatography on silica gel ( petroleum ether : etoac = 1 : 1 ) to obtain the titled product ( quantitively ). to a solution of 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - n -( 2 -( vinyloxy ) ethoxy )- 4 , 5 - dihydrothieno [ 3 , 2 - c ] pyridine - 7 - carboxamide ( 139 mg , 0 . 26 mmol ) in 5 ml meoh was added 1 . 5 ml 2 n hcl ( aq ). the reaction mixture was stirred for 30 min at room temperature . the reaction mixture was diluted with ch 2 cl 2 and h 2 o . the suspension was filtered and dried to give the product as white solid ( 54 mg , 40 %). 1 h nmr ( 400 mhz , dmso - d6 ) δ 11 . 31 ( s , 1h ), 8 . 30 ( s , 1h ), 7 . 62 - 7 . 63 ( d , j = 5 . 6 hz , 2h ), 7 . 56 - 7 . 59 ( d , j = 10 . 4 hz , 1h ), 7 . 49 - 7 . 50 ( d , j = 5 . 2 hz , 1h ), 7 . 30 - 7 . 32 ( d , j = 8 . 4 hz , 1h ), 6 . 49 - 6 . 53 ( t , j = 8 . 8 hz ), 4 . 68 ( s , 1h ), 3 . 60 ( m , 2h ), 3 . 43 ( m , 5h ); m / z = 504 [ m + 1 ] + , 442 [ m - och 2 ch 2 oh ] + , 427 [ m - nhoch 2 ch 2 oh ] + . following the same procedure as example 1 described , using 2 - fluoro - n -(( methylimino ) methylene )- 4 -( methylthio ) aniline as starting material to get the product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 90 ( s , 1h ), 9 . 86 ( s , 1h ), 7 . 50 ( d , j = 2 . 0 hz , 1h ), 7 . 05 - 6 . 98 ( m , 2h ), 6 . 96 ( d , j = 2 . 0 hz , 1h ), 6 . 79 ( t , j = 8 . 4 hz , 1h ), 4 . 16 - 4 . 09 ( m , 3h ), 3 . 79 - 3 . 75 ( m , 2h ), 3 . 28 ( s , 3h ), 2 . 48 ( s , 3h ); m / z = 331 [ m - nhoch 2 ch 2 oh ] + , 408 [ m + 1 ] + . following the same procedure as example 1 described , using 2 - chloro - n -(( methylimino ) methylene )- 4 -( methylthio ) aniline as starting material to get the product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 84 ( s , 1h ), 9 . 85 ( s , 1h ), 7 . 52 ( d , j = 2 . 4 hz , 1h ), 7 . 31 ( d , j = 2 . 4 hz , 1h ), 7 . 06 ( d , j = 6 . 0 hz , 1h ), 7 . 02 - 7 . 01 ( m , 1h ), 6 . 62 ( d , j = 8 . 4 hz , 1h ), 4 . 12 - 4 . 09 ( m , 3h ), 3 . 79 - 3 . 77 ( m , 2h ), 3 . 25 ( s , 3h ), 2 . 48 ( s , 3h ); m / z = 347 [ m - nhoch 2 ch 2 oh ] + , 424 [ m + 1 ] + . according to j . org . chem ., 1998 , 63 , 7680 - 7686 ., the title compound was made . 1 h nmr ( 400 mhz , cdcl 3 ): δ 4 . 10 ( s , 2h ), 3 . 90 ( s , 3h ), 3 . 75 ( s , 3h ), 2 . 49 ( s , 3h ). according to the procedure b , methyl 5 -( 2 - methoxy - 2 - oxoethyl )- 2 - methyloxazole - 4 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 93 ( s , 1h ), 7 . 49 - 7 . 52 ( dd , j = 1 . 8 & amp ; 9 . 8 hz , 1h ), 7 . 38 - 7 . 41 ( d , j = 8 . 4 hz , 1h ), 6 . 48 - 6 . 52 ( t , j = 8 . 4 hz , 1h ), 3 . 96 ( s , 3h ), 3 . 36 ( s , 3h ), 2 . 64 ( s , 3h ). following the same procedure as step c , example 1 described , the title product was obtained . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 86 ( s , 1h ), 9 . 51 ( s , 1h ), 7 . 46 ( d , j = 8 . 0 hz , 1h ), 7 . 42 ( d , j = 8 . 8 hz , 1h ), 6 . 59 ( t , j = 8 . 4 hz , 1h ), 4 . 13 - 4 . 11 ( m , 2h ), 3 . 92 - 3 . 89 ( m , 1h ), 3 . 81 - 3 . 77 ( m , 2h ), 3 . 31 ( s , 3h ), 2 . 68 ( s , 3h ); m / z = 503 [ m + 1 ] + , 441 [ m - och 2 ch 2 oh ] + , 426 [ m - nhoch 2 ch 2 oh ] + . according to j . chem . soc ., perkin trans . 1 , organic and bio - organic chemistry , 1999 , 7 , 765 - 776 , the title compound was made . 1 h nmr ( 400 mhz , cdcl 3 ): δ 4 . 13 ( s , 2h ), 3 . 86 ( s , 3h ), 3 . 75 ( s , 3h ), 2 . 47 ( s , 3h ). according to the procedure b , methyl 5 -( 2 - methoxy - 2 - oxoethyl )- 3 - methylisoxazole - 4 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 58 ( s , 1h ), 7 . 51 - 7 . 54 ( dd , j = 1 . 8 & amp ; 9 . 8 hz , 1h ), 7 . 44 - 7 . 46 ( d , j = 8 . 4 hz , 1h ), 6 . 63 - 6 . 67 ( t , j = 8 . 4 hz , 1h ), 3 . 98 ( s , 3h ), 3 . 23 ( s , 3h ), 2 . 60 ( s , 3h ). following the same procedure as step c , example 19 described , the title product was obtained . following the same procedure as step d , example 19 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 14 ( s , 1h ), 9 . 77 ( s , 1h ), 7 . 52 ( d , j = 9 . 6 hz , 1h ), 7 . 46 ( d , j = 8 . 4 hz , 1h ), 6 . 73 ( t , j = 8 . 0 hz , 1h ), 4 . 12 - 4 . 10 ( m , 2h ), 3 . 99 - 3 . 96 ( m , 1h ), 3 . 80 - 3 . 76 ( m , 2h ), 3 . 20 ( s , 3h ), 2 . 60 ( s , 3h ); m / z = 426 [ m - nhoch 2 ch 2 oh ] + . according to j . am . chem . soc , 1985 , 107 , 2196 - 2198 ., the title compound was made . 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 13 ( s , 1h ), 4 . 28 ( q , j = 7 . 2 hz , 2h ), 4 . 18 ( q , j = 6 . 8 hz , 2h ), 4 . 01 ( s , 2h ), 2 . 17 ( s , 3h ), 1 . 34 ( t , j = 6 . 8 hz , 3h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ). according to the procedure b , ethyl 2 -( 2 - ethoxy - 2 - oxoethyl )- 4 - methylfuran - 3 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 77 ( s , 1h ), 7 . 48 ( dd , j = 1 . 6 & amp ; 9 . 6 hz , 1h ), 7 . 35 ( d , j = 8 . 4 hz , 1h ), 7 . 28 ( q , j = 1 . 2 hz , 1h ), 6 . 42 ( t , j = 8 . 4 hz , 1h ), 4 . 40 ( q , j = 7 . 2 hz , 2h ), 3 . 34 ( s , 3h ), 2 . 37 ( d , j = 1 . 2 hz , 3h ), 1 . 41 ( t , j = 7 . 2 hz , 3h ); m / z = 471 [ m + 1 ] + . following the same procedure as step c , example 1 described , the title product was obtained via column chromatography purification . 1 h nmr ( 400 mhz , dmso - d6 ) δ 13 . 20 ( s , 1h ), 9 . 63 ( s , 1h ), 7 . 64 - 7 . 71 ( m , 2h ), 7 . 40 - 7 . 42 ( d , 1h ), 6 . 60 - 6 . 70 ( t , j = 8 . 8 hz , 1h ), 3 . 23 ( s , 3h ), 2 . 25 ( s , 3h ); m / z = 443 [ m + 1 ] + . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 78 ( s , 1h ), 9 . 86 ( s , 1h ), 7 . 47 - 7 . 49 ( dd , j = 1 . 6 & amp ; 9 . 6 hz , 1h ), 7 . 37 - 7 . 39 ( d , j = 8 . 4 hz , 1h ), 7 . 37 ( s , 1h ), 6 . 51 - 6 . 55 ( t , j = 8 . 4 hz , 1h ), 4 . 08 - 4 . 10 ( m , 2h ), 3 . 74 - 3 . 78 ( m , 2h ), 3 . 28 ( s , 3h ), 2 . 37 ( s , 3h ); m / z = 425 [ m - nhoch 2 ch 2 oh ] + , 502 [ m + 1 ] + . according to bioorganic & amp ; medicinal chemistry letters , 2008 , 18 ( 6 ), 2206 - 2210 ., the title compound was made . 1 h nmr ( 400 mhz , cdcl 3 ): δ 8 . 75 ( s , 1h ), 4 . 38 ( s , 2h ), 3 . 96 ( s , 3h ), 3 . 78 ( s , 3h ). according to the procedure b , methyl 5 -( 2 - methoxy - 2 - oxoethyl ) thiazole - 4 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the impure product . m / z = 460 [ m + 1 ]+. following the same procedure as step c , example 1 described , the title product was obtained via column chromatography purification . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cd 3 od ) δ 9 . 05 ( s , 1h ), 7 . 53 - 7 . 56 ( dd , j = 1 . 8 & amp ; 10 . 6 hz , 1h ), 7 . 38 - 7 . 40 ( d , j = 8 . 4 hz , 1h ), 6 . 61 ( s , 1h ), 3 . 59 - 3 . 71 ( m , 7h ); m / z = 505 [ m + 1 ] + , 443 [ m - och 2 ch 2 oh ] + . according to heterocycles , 1998 , 48 ( 5 ), 853 - 860 ., the title compound was made . 1 h nmr ( 400 mhz , cdcl 3 ): δ 4 . 30 ( s , 2h ), 3 . 94 ( s , 3h ), 3 . 76 ( s , 3h ), 2 . 72 ( s , 3h ). according to the procedure b , methyl 5 -( 2 - methoxy - 2 - oxoethyl )- 2 - methylthiazole - 4 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the impure product . m / z = 474 [ m + 1 ] + . following the same procedure as step c , example 1 described , the title product was obtained via column chromatography purification . 1 h nmr ( 400 mhz , dmso - d6 ) δ 13 . 20 ( s , 1h ), 9 . 63 ( s , 1h ), 7 . 68 ( dd , j = 1 . 6 hz & amp ; 10 . 4 hz , 1h ), 7 . 42 ( d , j = 8 . 4 hz , 1h ), 6 . 68 ( t , j = 8 . 4 hz , 1h ), 3 . 23 ( s , 3h ), 2 . 26 ( s , 3h ). following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cd 3 od ) 7 . 54 - 7 . 57 ( d , j = 10 . 4 hz , 1h ), 7 . 42 - 7 . 45 ( d , j = 9 . 2 hz , 1h ), 6 . 57 ( t , j = 8 . 4 hz , 1h ), 3 . 30 - 3 . 37 ( m , 7h ), 2 . 72 - 2 . 79 ( m , 3h ); m / z = 519 [ m + 1 ] + . according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 2 , 5 - dimethyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was coupling with ( r )— o -(( 2 , 2 - dimethyl - 1 , 3 - dioxolan - 4 - yl ) methyl ) hydroxylamine ( made according to tetrahedron letters , 2006 , 47 , 7607 - 7609 ), followed by treating with 2 n hcl in meoh to get the title product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 63 ( s , 1h ), 9 . 98 ( s , 1h ), 7 . 39 - 7 . 51 ( dd , j = 1 . 8 hz & amp ; 9 . 8 hz , 1h ), 7 . 37 - 7 . 39 ( d , j = 8 . 4 hz , 1h ), 6 . 58 ( s , 1h ), 6 . 48 - 6 . 53 ( t , j = 8 . 4 hz , 1h ), 4 . 42 - 4 . 43 ( d , j = 3 . 6 hz , 1h ), 4 . 04 - 4 . 15 ( m , 2h ), 3 . 97 - 3 . 99 ( t , j = 3 . 8 hz , 1h ), 3 . 74 - 3 . 78 ( m , 1h ), 3 . 64 - 3 . 68 ( m , 1h ), 3 . 30 ( s , 3h ), 2 . 48 ( s , 3h ), 2 . 37 - 2 . 40 ( t , j = 6 . 2 hz , 1h ); m / z = 532 [ m + 1 ] + , 425 [ m - nhoch 2 ch ( oh ) ch 2 oh ] + . dimethylamine ( 178 mg , 0 . 2 ml , 33 % aq ., 1 . 30 mmol ) was added to acetic acid ( 1 ml ) slowly at 0 ° c ., then 37 % aqueous formaldehyde ( formalin ) solution ( 0 . 1 ml , 1 . 33 mmol ) was added at 0 ° c . the mixture was then stirred at r . t ., and methyl 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylate ( 300 mg , 0 . 68 mmol ) was added at once . after addition of 20 ml thf , the mixture was heated to reflux . after consuming up of the methyl ester , water was added . using ch 2 cl 2 extracted , dried , filtered , concentrated the filtrate in vacuum , the residue was purified by flash column chromatography on silica gel to get the title product ( 255 mg , 75 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 77 ( s , 1h ), 7 . 49 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 37 ( d , j = 8 . 8 hz , 1h ), 6 . 76 ( s , 1h ), 6 . 44 ( t , j = 8 . 8 hz , 1h ), 3 . 94 ( s , 3h ), 3 . 63 ( s , 2h ), 3 . 36 ( s , 3h ), 2 . 35 ( s , 6h ); m / z = 455 [ m - n ( ch 3 ) 2 ] + . a mixture of methyl 2 -(( dimethylamino ) methyl )- 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylate ( 200 mg , 0 . 4 mmol ) and k 2 co 3 ( 166 mg , 1 . 2 mmol ) in 12 ml meoh : h 2 o ( 5 : 1 ) was heated at 70 ° c . for 2 h , the mixture was then concentrated in vacuum , the residue was then purified by flash column chromatography on silica gel ( ch 2 cl 2 : meoh = 10 : 1 to meoh ) to get the title product ( 180 mg , 93 %). 1 h nmr ( 400 mhz , dmso - d6 ) δ 7 . 65 ( dd , j = 2 . 0 & amp ; 10 . 8 hz , 1h ), 7 . 39 ( d , j = 8 . 4 hz , 1h ), 6 . 75 ( s , 1h ), 6 . 55 ( t , j = 8 . 4 hz , 1h ), 3 . 65 ( s , 2h ), 3 . 21 ( s , 3h ), 2 . 28 ( s , 6h ); m / z = 441 [ m - n ( ch 3 ) 2 ] + . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 77 ( s , 1h ), 10 . 69 ( s , 1h ), 7 . 46 - 7 . 49 ( dd , j = 1 . 8 & amp ; 9 . 8 hz , 1h ), 7 . 36 - 7 . 38 ( d , j = 8 . 4 hz , 1h ), 6 . 80 ( s , 1h ), 6 . 48 - 6 . 52 ( t , j = 8 . 4 hz , 1h ), 4 . 11 - 4 . 13 ( m , 2h ), 3 . 82 - 3 . 84 ( m , 2h ), 3 . 58 ( s , 2h ), 3 . 31 ( s , 3h ), 2 . 33 ( s , 6h ); m / z = 500 [ m - n ( ch 3 ) 2 ] + , 438 [ m - n ( ch 3 ) 2 — och 2 ch 2 oh ] + . following the same procedure as step a , example 28 described , using morpholine ( 1 . 5 eq .) instead of dimethylamine , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ): δ 9 . 76 ( s , 1h ), 7 . 49 ( dd , j = 2 . 0 & amp ; 10 . 0 hz , 1h ), 7 . 37 ( d , j = 8 . 4 hz , 1h ), 6 . 77 ( s , 1h ), 6 . 44 ( t , j = 8 . 4 hz , 1h ), 3 . 94 ( s , 3h ), 3 . 60 - 3 . 75 ( m , 6h ), 3 . 35 ( s , 3h ), 2 . 58 - 2 . 64 ( m , 4h ); m / z = 455 [ m - n ( ch 2 ch 2 och 2 ch 2 )] + , 564 [ m + na ] + . following the same procedure as step b , example 28 described , the title product was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 7 . 65 ( dd , j = 2 . 0 hz & amp ; 10 . 8 hz , 1h ), 7 . 39 ( dd , j = 1 . 2 & amp ; 8 . 4 hz , 1h ), 6 . 72 ( s , 1h ), 6 . 53 ( t , j = 8 . 4 hz , 1h ), 3 . 56 - 3 . 58 ( m , 6h ), 3 . 21 ( s , 3h ), 2 . 43 ( m , 4h ); m / z = 441 [ m - n ( ch 2 ch 2 och 2 ch 2 )] + . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 10 . 78 ( s , 1h ), 10 . 28 ( s , 1h ), 7 . 47 - 7 . 50 ( dd , j = 2 . & amp ; 10 . 6 hz , 1h ), 7 . 37 - 7 . 39 ( d , j = 8 . 4 hz , 1h ), 6 . 82 ( s , 1h ), 6 . 50 - 6 . 54 ( t , j = 8 . 4 hz , 1h ), 4 . 11 - 4 . 13 ( m , 2h ), 3 . 82 - 3 . 84 ( m , 2h ), 3 . 73 - 3 . 75 ( t , j = 4 . 4 hz , 4h ), 3 . 73 ( s , 2h ), 3 . 30 ( s , 3h ), 2 . 54 ( s , 4h ); m / z = 500 [ m - n ( ch 2 ch 2 och 2 ch 2 )] + , 438 [ m - n ( ch 2 ch 2 och 2 ch 2 )— och 2 ch 2 oh ] + . the title compound was made according to j . org . chem ., 1988 , 53 , 3457 - 3465 . 1 h nmr ( 400 mhz , cdcl 3 ): δ 4 . 54 ( s , 2h ), 0 . 91 ( s , 9h ), 0 . 12 ( s , 6h ). to a stirred solution of 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxamide ( 1 . 0 eq .) in anhydrous thf was added nah ( 1 . 1 eq ., 60 %) portionwise at 0 ° c ., the mixture was then stirred at room temperature for 1 h , then 2 -( tert - butyldimethylsilyloxy ) acetyl chloride ( 1 . 5 eq .) in thf was added slowly with dropping funnel , the mixture was then stirred at room temperature for 3 days . after the addition of water and ethyl acetate , the water layer was extracted with ethyl acetate , washed with sat . nacl , dried over mgso 4 , filtered , concentrated in vacuum , the residue was purified by column chromatography on silica gel to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 24 ( s , 1h ), 10 . 88 ( s , 1h ), 7 . 53 ( d , j = 2 . 4 hz , 1h ), 7 . 49 ( dd , j = 1 . 6 & amp ; 10 . 0 hz , 1h ), 7 . 42 ( d , j = 8 . 4 hz , 1h ), 7 . 03 ( d , j = 2 . 4 hz , 1h ), 6 . 61 ( t , j = 8 . 4 hz , 1h ), 4 . 34 ( s , 2h ), 3 . 29 ( s , 3h ), 0 . 97 ( s , 9h ), 0 . 20 ( s , 6h ); m / z = 600 [ m + 1 ] + . to a solution of n -( 2 -( tert - butyldimethylsilyloxy ) acetyl )- 6 -( 2 - fluoro - 4 - iodophenyl amino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxamide ( 30 mg , 0 . 05 mmol ) in meoh ( 5 ml ) was added 2n hcl ( 1 ml ). the mixture was then stirred at room temperature for 5 min . the white precipitate was filtered , dried , afforded the title compound ( 6 . 7 mg , 28 %). 1 h nmr ( 400 mhz , dmso - d6 ) δ 10 . 83 ( s , 1h ), 9 . 21 ( s , 1h ), 7 . 92 ( d , j = 2 . 4 hz , 1h ), 7 . 63 ( dd , j = 2 . 0 & amp ; 10 . 8 hz , 1h ), 7 . 37 ( d , j = 8 . 4 hz , 1h ), 7 . 01 ( d , j = 2 . 0 hz , 1h ), 6 . 68 ( t , j = 8 . 8 hz , 1h ), 5 . 45 ( t , j = 6 . 0 hz , 1h ), 4 . 06 ( t , j = 6 . 0 hz , 2h ), 3 . 35 ( s , 3h ); m / z = 486 [ m + 1 ] + . to a neat diethyl 3 - oxopentanedioate ( 20 . 2 g , 0 . 10 mol ) was added sulfurous dichloride ( 13 . 5 g , 0 . 11 mol ) dropwise at 0 ° c ., the mixture was then stirred at rt for 1 h followed by heated at 50 ° c . for 0 . 5 h . the resulting solution was added dropwise to a solution of thiourea ( 7 . 6 g , 1 . 0 mol ) in etoh ( 50 ml ). the mixture was reflux for 0 . 5 h and added into ice water then alkaline with aq . na 2 co 3 , the precipitate was filtered off and dried to afford the title compound ( 16 g , 62 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 6 . 01 ( brs , 2h ), 4 . 27 ( q , j = 7 . 2 hz , 2h ), 4 . 17 ( q , j = 7 . 2 hz , 2h ), 4 . 03 ( s , 2h ), 1 . 34 ( t , j = 7 . 2 hz , 3h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ). to a solution of ethyl 2 - amino - 4 -( 2 - ethoxy - 2 - oxoethyl ) thiazole - 5 - carboxylate ( 4 . 0 g , 15 . 5 mmol ) and ( boc ) 2 o ( 13 . 5 g , 61 . 9 mmol ) in thf ( 50 ml ) was added et 3 n ( 9 . 4 ml , 61 . 9 mmol ) and dmap ( 190 mg , 1 . 55 mmol ) at rt , the mixture was then heated under reflux overnight . after quenching with water , the mixture was partitioned between dichloromethane and saturated nacl . the organic layers was separated , dried , concentrated under reduced pressure , residue was purified by column chromatography on silica gel ( petroleum ether : etoac = 2 : 1 ) to obtain the titled compound ( 1 . 2 g , 17 %) and ethyl 2 -( tert - butoxycarbonylamino )- 4 -( 2 - ethoxy - 2 - oxoethyl ) thiazole - 5 - carboxylate ( 1 . 0 g , 18 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 4 . 32 ( q , j = 7 . 2 hz , 2h ), 4 . 19 ( s , 2h ), 4 . 16 ( q , j = 7 . 2 hz , 2h ), 1 . 44 ( s , 9h ), 1 . 39 ( s , 9h ), 1 . 36 ( t , j = 7 . 2 hz , 3h ), 1 . 24 ( t , j = 7 . 2 hz , 3h ). according to the procedure b , ethyl 2 -( bis ( tert - butoxycarbonyl ) amino )- 4 -( 2 - ethoxy - 2 - oxoethyl ) thiazole - 5 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product via column chromatography purification . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 49 ( s , 1h ), 7 . 48 - 7 . 51 ( dd , j = 9 . 8 & amp ; 1 . 8 hz , 1h ), 7 . 35 - 7 . 37 ( d , j = 8 . 4 hz , 1h ), 6 . 42 - 6 . 46 ( t , j = 8 . 4 hz , 1h ), 4 . 37 ( q , j = 7 . 2 hz , 2h ), 3 . 42 ( s , 3h ), 1 . 54 ( s , 9h ), 1 . 43 ( s , 9h ), 1 . 38 ( t , j = 7 . 2 hz , 3h ). following the same procedure as step c , example 1 described , the title product was obtained via column chromatography purification . following the same procedure ( coupling section ) as step d , example 1 described , the title product was obtained via column chromatography purification . 1 h nmr ( 400 mhz , cdcl 3 ) δ 12 . 05 ( s , 1h ), 11 . 45 ( s , 1h ), 7 . 47 - 7 . 52 ( dd , j = 10 . 0 & amp ; 2 . 0 hz , 1h ), 7 . 37 - 7 . 39 ( d , j = 8 . 4 hz , 1h ), 6 . 55 ( t , j = 8 . 4 hz , 1h ), 6 . 50 ( dd , j = 14 . 0 & amp ; 6 . 8 hz , 1h ), 4 . 27 - 4 . 29 ( m , 2h ), 4 . 20 ( dd , j = 14 . 4 & amp ; 2 . 4 hz , 1h ), 4 . 04 ( dd , j = 6 . 8 & amp ; 2 . 4 hz , 1h ), 3 . 96 - 3 . 99 ( m , 2h ), 3 . 35 ( s , 3h ), 1 . 50 ( s , 9h ), 1 . 43 ( s , 9h ). following the same procedure as step d , example 1 described , after treating with 2 n hcl in meoh , the solution was concentrated in vacuum , the residue was then dissolved in dichloromethane / trifluoroacetic acid solution ( v / v , 1 / 1 ) and stirred for 1 h , then concentrated under reduced pressure , the residue was purified by column chromatography on silica gel to afford the title product . 1 h nmr ( 400 mhz , dmso - d6 ) δ 12 . 24 ( s , 1h ), 10 . 56 ( s , 1h ), 8 . 43 ( s , 2h ), 7 . 68 ( dd , j = 10 . 4 & amp ; 2 . 0 hz , 1h ), 7 . 42 ( d , j = 7 . 6 hz , 1h ), 6 . 65 ( t , j = 8 . 4 hz , 1h ), 4 . 71 ( t , j = 5 . 6 hz , 1h ), 3 . 84 ( t , j = 4 . 8 hz , 1h ), 3 . 53 - 3 . 57 ( m , 2h ), 3 . 20 ( s , 3h ); m / z = 520 [ m + 1 ] + . according to the same procedure as step b , example 31 described , the title compound was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 11 . 99 ( brs , 1h ), 4 . 22 ( q , j = 7 . 2 hz , 2h ), 4 . 07 ( q , j = 7 . 2 hz , 2h ), 4 . 00 ( s , 2h ), 1 . 49 ( s , 9h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ), 1 . 17 ( t , j = 7 . 2 hz , 3h ). to a solution of ethyl 2 -( tert - butoxycarbonylamino )- 4 -( 2 - ethoxy - 2 - oxoethyl ) thiazole - 5 - carboxylate ( 50 mg , 1 . 0 eq .) in acetone ( 5 ml ) was added mei ( 23 . 8 mg , 1 . 2 eq .) and k 2 co 3 ( 19 . 2 mg , 1 . 0 eq .) at 0 ° c . the mixture was warmed to room temperature and stirred for 3 . 5 h at this temperature . the solvent was removed under reduced pressure and the residue was purified by column chromatography on silica gel ( etoac / petroleum ether = 1 / 20 ) to obtain the titled compound ( 50 mg , yield = 98 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 4 . 28 ( q , j = 7 . 2 hz , 2h ), 4 . 18 ( q , j = 7 . 2 hz , 2h ), 4 . 11 ( s , 2h ), 3 . 53 ( s , 3h ), 1 . 58 ( s , 9h ), 1 . 33 ( t , j = 7 . 2 hz , 3h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ). according to the procedure b , ethyl 2 -( tert - butoxycarbonyl ( methyl ) amino )- 4 -( 2 - ethoxy - 2 - oxoethyl ) thiazole - 5 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 48 ( s , 1h ), 7 . 48 ( dd , j = 10 . 0 & amp ; 2 . 0 hz , 1h ), 7 . 34 ( d , j = 8 . 4 hz , 1h ), 6 . 39 ( t , j = 8 . 4 hz , 1h ), 4 . 37 ( q , j = 7 . 2 hz , 2h ), 3 . 65 ( s , 3h ), 3 . 41 ( s , 3h ), 1 . 56 ( s , 9h ), 1 . 41 ( t , j = 7 . 2 hz , 3h ). following the same procedure as step c , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 10 . 36 ( brs , 1h ), 9 . 28 ( brs , 1h ), 7 . 74 ( d , j = 10 . 4 hz , 1h ), 7 . 49 ( d , j = 8 . 0 hz , 1h ), 6 . 85 ( t , j = 8 . 8 hz , 1h ), 3 . 15 ( s , 3h ), 2 . 99 ( s , 3h ). following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cd 3 od ) δ 7 . 40 ( dd , j = 10 . 8 & amp ; 2 . 0 hz , 1h ), 7 . 27 ( d , j = 8 . 4 hz , 1h ), 6 . 35 ( t , j = 8 . 8 hz , 1h ), 3 . 81 - 3 . 83 ( m , 2h ), 3 . 59 - 3 . 61 ( m , 2h ), 3 . 31 ( s , 3h ), 2 . 95 ( s , 3h ); m / z = 534 [ m + 1 ] + , 457 [ m - nhoch 2 ch 2 oh ] + , 556 [ m + na ] + . to a neat diethyl 3 - oxopentanedioate ( 5 . 0 g , 25 mmol ) was added sulfurous dichloride ( 2 ml , 25 mol ) dropwise at 0 ° c ., the mixture was then stirred at rt for 2 h followed by heated at 50 ° c . for 0 . 5 h . the resulting solution was added dropwise to a solution of methanethioamide ( 1 . 51 g , 25 mol ) in etoh ( 15 ml ). the mixture was reflux for 0 . 5 h and added into ice water , the precipitate was filtered off and dried to afford the title compound ( 2 g , 33 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 8 . 82 ( s , 1h ), 4 . 35 ( q , j = 7 . 2 hz , 2h ), 4 . 26 ( s , 2h ), 4 . 19 ( q , j = 7 . 2 hz , 2h ), 1 . 37 ( t , j = 7 . 2 hz , 3h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ). according to the procedure b , ethyl 4 -( 2 - ethoxy - 2 - oxoethyl ) thiazole - 5 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product via column chromatography purification . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 50 ( s , 1h ), 9 . 19 ( s , 1h ), 7 . 50 ( dd , j = 10 . 0 & amp ; 2 . 0 hz , 1h ), 7 . 37 ( d , j = 8 . 4 hz , 1h ), 6 . 44 ( t , j = 8 . 4 hz , 1h ), 4 . 46 ( q , j = 7 . 2 hz , 2h ), 3 . 44 ( s , 3h ), 1 . 41 ( t , j = 7 . 2 hz , 1h ). following the same procedure as step c , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 13 . 20 ( brs , 1h ), 9 . 65 ( s , 1h ), 7 . 66 - 7 . 69 ( d , j = 11 . 2 hz , 1h ), 7 . 40 - 7 . 42 ( d , j = 8 . 0 hz , 1h ), 6 . 72 - 6 . 74 ( t , j = 8 . 8 hz , 1h ), 3 . 36 ( s , 3h ). following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cdcl 3 ) δ 12 . 36 ( s , 1h ), 11 . 36 ( s , 1h ), 9 . 15 ( s , 1h ), 7 . 48 - 7 . 51 ( d , j = 9 . 6 hz , 1h ), 7 . 40 - 7 . 42 ( d , j = 8 . 4 hz , 1h ), 6 . 60 ( t , j = 8 . 4 hz , 1h ), 4 . 08 - 4 . 10 ( m , 2h ), 3 . 76 - 3 . 77 ( m , 2h ), 3 . 36 ( s , 3h ); m / z = 505 [ m + 1 ] + , 527 [ m + na ] + , 428 [ m - nhoch 2 ch 2 oh ] + . sulfuryl dichloride ( 2 ml , 1 . 0 eq .) was added dropwise to diethyl 3 - oxopentanedioate ( neat , 5 g , 1 . 0 eq .) at 0 ° c . the mixture was stirred for 2 h at room temperature , and heated for 30 min at 50 ° c . the resulting solution was added dropwise to a solution of ammonium dithiocarbamate ( 2 . 3 g , 1 . 0 eq .) in etoh ( 50 ml ). the mixture was heated at 50 ° c . for at least 4 h until the precipitate was formed . the mixture was cooled and the solid was filtered off . the filtration was concentrated and purified by column chromatography ( etoac / petroleum ether = 1 / 5 ) to obtain the titled compound ( 1 g , 14 . 7 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 50 ( brs , 1h ), 4 . 22 - 4 . 32 ( m , 4h ), 4 . 10 ( s , 2h ), 1 . 29 - 1 . 36 ( m , 6h ). to a solution of ethyl 4 -( 2 - ethoxy - 2 - oxoethyl )- 2 - mercaptothiazole - 5 - carboxylate ( 356 mg , 1 . 0 eq .) in acetone ( 10 ml ) was added mei ( 220 mg , 1 . 2 eq .) and k 2 co 3 ( 150 mg , 1 . 0 eq .) at 0 ° c . after warming to room temperature and stirring for 30 min , the solvent was removed in vacuum and the residue was purified by flash column chromatography ( etoac / petroleum ether = 1 / 20 ) to obtain the titled compound ( 150 mg , 50 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 4 . 30 ( q , j = 7 . 2 hz , 2h ), 4 . 20 ( q , j = 7 . 2 hz , 2h ), 4 . 17 ( s , 2h ), 2 . 69 ( s , 3h ), 1 . 34 ( t , j = 7 . 2 hz , 3h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ). according to the procedure b , ethyl 4 -( 2 - ethoxy - 2 - oxoethyl )- 2 -( methylthio ) thiazole - 5 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product by column chromatography . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 57 ( s , 1h ), 7 . 48 ( dd , j = 10 . 0 & amp ; 1 . 6 hz , 1h ), 7 . 36 ( d , j = 8 . 8 hz , 1h ), 6 . 42 ( t , j = 8 . 4 hz , 1h ), 4 . 38 ( q , j = 7 . 2 hz , 2h ), 3 . 40 ( s , 3h ), 2 . 81 ( s , 3h ), 1 . 41 ( t , j = 7 . 2 hz , 3h ). following the same procedure as step c , example 1 described , the title product was obtained . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cd 3 od ) δ 7 . 49 ( d , j = 10 . 0 hz , 1h ), 7 . 37 ( d , j = 6 . 8 hz , 1h ), 6 . 59 ( t , j = 8 . 4 hz , 1h ), 3 . 91 ( m , 2h ), 3 . 66 ( m , 2h ), 3 . 29 ( s , 3h ), 2 . 77 ( s , 3h ); m / z = 551 [ m + 1 ] + , 573 [ m + na ] + , 474 [ m - nhoch 2 ch 2 oh ] + . according to procedure c , 6 -( 2 - chloro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was reacted with 3 - methoxypropan - 1 - amine then treated with bbr 3 at 0 ° c . to get the desired product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 11 . 19 ( s , 1h ), 7 . 77 ( d , j = 2 . 0 hz , 1h ), 7 . 68 ( m , 1h ), 7 . 51 ( d , j = 2 . 4 hz , 1h ), 7 . 43 ( dd , j = 8 . 4 & amp ; 2 . 0 hz , 1h ), 7 . 02 ( d , j = 2 . 4 hz , 1h ), 6 . 34 ( d , j = 8 . 4 hz , 1h ), 3 . 70 - 3 . 72 ( m , 2h ), 3 . 61 - 3 . 65 ( m , 2h ), 3 . 30 ( s , 3h ), 2 . 60 ( brs , 1h ), 1 . 81 - 1 . 87 ( m , 2h ); m / z = 502 [ m + 1 ] + , 427 [ m - nhch 2 ch 2 ch 2 oh ] + . according to the procedure b , ethyl 2 -( 2 - ethoxy - 2 - oxoethyl )- 4 - methylfuran - 3 - carboxylate was reacted with 2 - chloro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product by column chromatography . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 69 ( s , 1h ), 7 . 77 ( d , j = 1 . 6 hz , 1h ), 7 . 43 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 7 . 29 ( q , j = 1 . 2 hz , 1h ), 6 . 31 ( d , j = 8 . 4 hz , 1h ), 4 . 40 ( q , j = 6 . 8 hz , 2h ), 3 . 31 ( s , 3h ), 2 . 37 ( d , j = 1 . 2 hz , 3h ), 1 . 41 ( t , j = 6 . 8 hz , 3h ). following the same procedure as step c , example 1 described , the title product was obtained . following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 11 . 26 ( s , 1h ), 8 . 96 ( s , 1h ), 7 . 78 ( d , j = 2 . 0 hz , 1h ), 7 . 68 ( s , 1h ), 7 . 47 ( dd , j = 2 . 0 & amp ; 8 . 4 hz , 1h ), 6 . 45 ( d , j = 8 . 4 hz , 1h ), 4 . 64 ( t , j = 5 . 6 hz , 1h ), 3 . 73 ( t , j = 8 . 8 hz , 2h ), 3 . 51 ( dt , j = 8 . 8 & amp ; 5 . 6 hz , 2h ), 3 . 26 ( s , 3h ), 2 . 27 ( s , 3h ); m / z = 441 [ m - nhoch 2 ch 2 oh ] + , 518 [ m + 1 ] + . sulfuryl dichloride ( 4 ml , 1 . 0 eq .) was added dropwise to diethyl 3 - oxopentanedioate ( neat , 10 g , 1 . 0 eq .) at 0 ° c . the mixture was stirred for 2 h at room temperature , and heated for 30 min at 50 ° c . the resulting solution was added dropwise to a solution of ethanethioamide ( 3 . 7 g , 1 . 0 eq .) in etoh ( 100 ml ). the mixture was heated at 50 ° c . for at least 4 h until the precipitate was formed . the mixture was cooled and the solid was filtered off . the filtration was concentrated and purified by column chromatography ( etoac / petroleum ether = 1 / 20 ) to obtain the titled compound ( 5 . 9 g , 46 . 4 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 4 . 31 ( q , j = 7 . 2 hz , 2h ), 4 . 18 ( q , j = 7 . 2 hz , 2h ), 4 . 17 ( s , 2h ), 2 . 70 ( s , 3h ), 1 . 34 ( t , j = 7 . 2 hz , 3h ), 1 . 26 ( t , j = 7 . 2 hz , 3h ). according to the procedure b , ethyl 4 -( 2 - ethoxy - 2 - oxoethyl )- 2 - methylthiazole - 5 - carboxylate was reacted with 2 - fluoro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product by column chromatography . 1 h nmr ( 400 mhz , dmso - d6 ) δ 8 . 62 ( s , 1h ), 7 . 59 ( dd , j = 10 . 8 & amp ; 1 . 6 hz , 1h ), 7 . 34 ( d , j = 8 . 4 hz , 1h ), 6 . 60 ( t , j = 8 . 8 hz , 1h ), 3 . 96 ( q , j = 7 . 2 hz , 2h ), 3 . 44 ( s , 3h ), 2 . 79 ( s , 3h ), 1 . 02 ( t , j = 7 . 2 hz , 3h ). following the same procedure as step c , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 13 . 30 ( brs , 1h ), 9 . 60 ( s , 1h ), 7 . 67 ( dd , j = 10 . 4 & amp ; 1 . 2 hz , 1h ), 7 . 42 ( d , j = 8 . 4 hz , 1h ), 6 . 76 ( t , j = 8 . 4 hz , 1h ), 3 . 29 ( s , 3h ), 2 . 86 ( s , 3h ). following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cd 3 od ) δ 7 . 49 ( dd , j = 10 . 4 & amp ; 1 . 6 hz , 1h ), 7 . 37 ( d , j = 8 . 4 hz , 1h ), 6 . 57 ( t , j = 8 . 4 hz , 1h ), 3 . 93 ( t , j = 4 . 4 hz , 2h ), 3 . 66 ( t , j = 4 . 4 hz , 2h ), 3 . 28 ( s , 3h ), 2 . 80 ( s , 3h ); m / z = 442 [ m - nhoch 2 ch 2 oh ] + , 519 [ m + 1 ] + . according to j . org . chem ., 1990 , 55 , 2578 - 2580 and wo2007 / 044515 , tert - butyl piperidine - 1 - carboxylate was ′- lithiation with sec - buli in ethyl ether , followed by reacting with benzyl 3 - oxoazetidine - 1 - carboxylate to afford the title compound ( yield = 11 %). 1 h nmr ( 400 mhz , cdcl 3 ) δ 7 . 27 - 7 . 37 ( m , 5h ), 5 . 40 ( brs , 1h ), 5 . 09 ( s , 2h ), 4 . 03 ( d , j = 9 . 2 hz , 1h ), 3 . 94 ( d , j = 9 . 2 hz , 1h ), 3 . 82 - 3 . 86 ( m , 3h ), 3 . 49 ( brs , 1h ), 3 . 06 ( brs , 1h ), 1 . 90 - 1 . 95 ( m , 1h ), 1 . 50 - 1 . 63 ( m , 5h ), 1 . 44 ( s , 9h ). tert - butyl 2 -( 1 -( benzyloxycarbonyl )- 3 - hydroxyazetidin - 3 - yl ) piperidine - 1 - carboxylate and 10 % pd / c were taken into methanol and the mixture hydrogenated at ambient pressure for 1 h . after filtration , the filtrate was concentrated to get the title product . 1 h nmr ( 400 mhz , cd 3 od ) δ 4 . 29 - 4 . 32 ( m , 1h ), 3 . 85 ( brs , 1h ), 3 . 72 ( d , j = 8 . 8 hz , 1h ), 3 . 54 ( d , j = 9 . 2 hz , 1h ), 3 . 42 ( d , j = 9 . 6 hz , 1h ), 3 . 39 ( d , j = 9 . 2 hz , 1h ), 3 . 35 - 3 . 42 ( brs , 1h ), 1 . 90 - 1 . 96 ( m , 1h ), 1 . 73 - 1 . 78 ( m , 1h ), 1 . 54 - 1 . 63 ( m , 4h ), 1 . 47 ( s , 9h ). according to procedure c , 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carboxylic acid was coupling with tert - butyl 2 -( 3 - hydroxyazetidin - 3 - yl ) piperidine - 1 - carboxylate to obtain the title product . 1 h nmr ( 400 mhz , cdcl 3 ) δ 9 . 01 ( brs , 1h ), 7 . 47 ( d , j = 1 . 6 hz , 1h ), 7 . 46 ( dd , j = 10 . 0 & amp ; 1 . 6 hz , 1h ), 7 . 34 ( d , j = 8 . 8 hz , 1h ), 6 . 99 ( d , j = 1 . 6 hz , 1h ), 6 . 35 ( t , j = 8 . 8 hz , 1h ), 3 . 78 - 4 . 19 ( m , 6h ), 3 . 43 ( s , 3h ), 1 . 55 - 3 . 12 ( m , 8h ), 1 . 41 ( s , 9h ). tert - butyl 2 -( 1 -( 6 -( 2 - fluoro - 4 - iodophenylamino )- 5 - methyl - 4 - oxo - 4 , 5 - dihydrofuro [ 3 , 2 - c ] pyridine - 7 - carbonyl )- 3 - hydroxyazetidin - 3 - yl ) piperidine - 1 - carboxylate was dissolved in the solution of 4n hcl in dioxane , then heated under reflux to obtain the product . 1 h nmr ( 400 mhz , dmso - d6 ) δ 8 . 51 ( brs , 1h ), 7 . 91 ( d , j = 2 . 0 hz , 1h ), 7 . 58 ( d , j = 10 . 8 hz , 1h ), 7 . 34 ( d , j = 9 . 2 hz , 1h ), 7 . 00 ( d , j = 2 . 0 hz , 1h ), 6 . 61 ( t , j = 9 . 2 hz , 1h ), 5 . 60 ( brs , 1h ), 3 . 65 - 3 . 88 ( m , 3h ), 3 . 48 - 3 . 53 ( m , 1h ), 3 . 39 ( s , 3h ), 2 . 88 - 2 . 92 ( m , 1h ), 2 . 42 - 2 . 49 ( m , 1h ), 2 . 26 - 2 . 33 ( m , 1h ), 1 . 60 - 1 . 70 ( m , 2h ), 1 . 40 - 1 . 50 ( m , 4h ); m / z = 567 [ m + 1 ] + . according to the procedure b , ethyl 4 -( 2 - ethoxy - 2 - oxoethyl )- 2 -( methylthio ) thiazole - 5 - carboxylate was reacted with 2 - chloro - 4 - iodo - n -(( methylimino ) methylene ) aniline to obtain the desired product by column chromatography . 1 h nmr ( 400 mhz , dmso - d6 ) δ 8 . 65 ( brs , 1h ), 7 . 77 ( d , j = 1 . 6 hz , 1h ), 7 . 46 ( dd , j = 8 . 4 & amp ; 1 . 6 hz , 1h ), 6 . 57 ( d , j = 8 . 4 hz , 1h ), 4 . 02 ( q , j = 7 . 2 hz , 2h ), 3 . 36 ( s , 3h ), 2 . 79 ( s , 3h ), 1 . 08 ( t , j = 7 . 2 hz , 3h ). following the same procedure as step c , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , dmso - d6 ) δ 13 . 10 ( brs , 1h ), 9 . 58 ( brs , 1h ), 7 . 84 ( d , j = 2 . 0 hz , 1h ), 7 . 51 ( dd , j = 8 . 8 & amp ; 2 . 0 hz , 1h ), 6 . 65 ( d , j = 8 . 8 hz , 1h ), 3 . 22 ( s , 3h ), 2 . 81 ( s , 3h ). following the same procedure as step d , example 1 described , the title product was obtained . 1 h nmr ( 400 mhz , cd 3 od ) δ 7 . 75 ( d , j = 1 . 6 hz , 1h ), 7 . 47 ( dd , j = 8 . 4 & amp ; 2 . 0 hz , 1h ), 6 . 47 ( d , j = 8 . 4 hz , 1h ), 3 . 92 ( t , j = 4 . 8 hz , 2h ), 3 . 66 ( t , j = 4 . 8 hz , 2h ), 3 . 24 ( s , 3h ), 2 . 78 ( s , 3h ); m / z = 490 [ m - nhoch 2 ch 2 oh ] + , 567 [ m + 1 ] + . the kinase glo plus assay kit was purchased from promega . the substrate , apt , dtt , and dimethylsulfoxide were purchased from sigma - aldrich . the map2k1 ( mek1 ) kinase , europium labeled antibody , tracer 236 and binding buffer a were purchased from invitrogen . the recombinant human epithelial growth factor ( egf ) was purchased from r & amp ; d system . the surefire phospho - erk1 / 2 assay kit and the alphascreen general igg ( protein a ) detection kit were both purchased from perkinelmer . compounds were diluted from dmso stocks into 1 × buffer ( 20 mm mops , ph 7 . 4 , 5 mm mgcl 2 , 0 . 5 mm mncl 2 , 100 um sodium orthovanadate , 0 . 01 % triton x - 100 , 1 mm dtt ). a typical reaction assay contained 0 . 01 nanomoles mek1 kinase , 0 . 01 nanomoles atp , 10 nanograms substrate . the screening assay essentially comprised four additions . 2 ul of diluted compounds were dispensed to 384 well white assay plates . 6 ul of kinase - substrate cocktail was then added to each well . 2 ul 5 × atp was subsequently added to each well to start the reaction . a top seal was applied and the plate was incubated at 22 degree avoiding light for 60 minutes . finally , 10 ul of the kinase glo plus reagent was added to each well to stop the reaction . incubated at room temperature and avoid light for ten minutes . the top seal was removed and the plate was counted by the envision 2104 multi labeled plate reader ( perkinelmer ) with a standard luminescent program . the intensity of luminescent signal was quantitated and this data was used to generate dose response curves and ic 50 calculations by the prism program . effects of compounds in the cell were determined by the alphascreen assay for phosphorylated erk . human a375 melanoma cancer cells were plated in a 96 wells plate at 80 , 000 cells per well and grown in a 37 degree humidified co 2 incubator . the following day , cells were treated with a range of compound concentrations for 60 minutes at 37 degree . the cells were then lysed and 4 ul of each lysate were transferred into the 384 well white reaction plate . a cocktail of alphascreen beads and buffer was freshly prepared and this mixture was dispensed into each well in a room with low light . a top seal was applied and the plate was incubated at 25 degree avoiding light for two hours . the top seal was removed when the plate was counted by the envision multi labeled plate reader ( perkinelmer ) with an optimized alphascreen program . the intensity of signal was quantitated and this data was used to generate dose response curves and ic 50 calculations by the prism program . select compounds prepared as described above were assayed according to the biological procedures described herein . the results are given in the table below :

the invention provides novel substituted 6 - arylamino pyridone carboxamides represented by formula i , or a pharmaceutically acceptable salt , solvate , poly - morph , ester , tautomer or prodrug thereof , and a composition comprising these compounds . the compounds provided can be used as inhibitors of mek and are useful in the treatment of inflammatory diseases , cancer and other hyperproliferative diseases . the invention further provides a method of treatment for inflammatory diseases , cancer and other hyperproliferative diseases in mammals , especially humans .

the present invention relates to methods for commercial scale production under an inert atmosphere of a transdermal formulation comprising a therapeutically effective amount of a hormone by dissolving a polyalcohol in an amount between 1 % and 10 % by weight of the formulation and a permeation enhancer in an amount between 1 % and 30 % by weight of the formulation in an alkanol in an inert atmosphere to form a stirred solution ; adding the therapeutically effective amount of the hormone to the stirred solution to form a hormone solution ; adding purified water to the hormone solution to form a hydroalcoholic mixture ; adding to the hydroalcoholic mixture a sequestering agent in an amount between 0 . 03 % and 0 . 09 % by weight of the formulation to form a second solution ; adding a gelling agent to the second solution in an amount between 1 % and 10 % by weight of the formulation to form a third solution having viscosity between 16 , 000 and 40 , 000 cps ; adding ph regulator in an amount between 0 . 2 % and 0 . 5 % by weight of the formulation to the third solution to adjust the ph to between 5 and 7 to form the transdermal testosterone formulation ; and collecting the transdermal testosterone formulation . unless defined otherwise , all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which this invention belongs . all patents and publications referred to herein are incorporated by reference . as used herein , “ ameliorate ” refers to any lessening , whether permanent or temporary , lasting or transient that can be attributed to or associated with administration of the testosterone formulation . as used herein , a “ commercial scale ” means that the method for producing a transdermal testosterone formulation is suitable for producing at least 500 kilograms of the transdermal testosterone formulation . as used herein , “ dose ” and “ dosage ” mean a specific amount of a hormone for administration . as used herein , “ estrogen ” or “ estrogens ” has its conventional meaning and comprises estrogen and estrogen derivatives such as 17 - beta - estradiol and esters thereof , ethinylestradiol , estriol ( trihydroxyestrin ), estrone , conjugated estrogens , in particular premarin , sodium estrone sulfate , 8 ( 9 )- dehydroestradiol derivatives , 17alfa - dihydroequilin , equilenin , 17alfa - dihydroequilenin , esterified estrogens , and equilin . as used herein , “ hormone ” or “ hormones ” “ has its conventional meaning and comprises an estrogen , a progesterone , or a testosterone compound . as used herein , “ progesterone ” has its conventional meaning and comprises progesterone , norethisterone acetate , norgestrel , levonorgestrel , gestodene , cpa , chlormadinone acetate , drospirorenone , and 3 - ketodesogestrel . as used herein , “ a testosterone compound ” means a compound selected from the group consisting of testosterone ( 17 - β - hydroxyandrostenone ), testosterone enanthate , testosterone propionate , testosterone decanoate , testosterone cypionate , methyl testosterone , testolactone , oxymetholone , fluoxymesterone and enanthate , propionate , cypionate , phenylacetate , acetate , isobutyrate , buciclate , heptanoate , decanoate , undecanoate , caprate and isocaprate esters of testosterone and 4 - dihydrotestosterone . as used herein , “ therapeutically effective amount ” means a sufficient amount or dose to provide the desired therapeutic effect . as used herein , “ transdermal formulation ” means a formulation for transdermal administration , i . e ., delivery by passage of a hormone such as a testosterone through the skin and into the bloodstream . the methods of the present invention for commercial scale production of a transdermal hormone formulation are suitable for producing batches of at least 500 kilograms of the transdermal hormone formulation . in certain configurations , the instant methods have the capacity to produce greater than 1 , 000 kilograms or even 1 , 200 kilograms of the transdermal hormone formulation . there are a number of reaction vessels suitable for use in the methods herein . in one embodiment , the manufacturing of transdermal hormone formulations at batch quantities at or exceeding 500 kilograms is performed in two separate reaction vessels . the first vessel is typically a hemispherical , jacketed manufacturing vessel having a capacity of about 700 liters and is used for the preparation of a primary compounding solution . preferably , this vessel is made of stainless steel . in one embodiment , the second vessel is a round bottom , jacketed , pressure / vacuum manufacturing vessel having a capacity of about 1350 liters and is used for ph adjustment , final mixing , deaeration , and gel formation . preferably , this vessel is made of stainless steel , and is capable of maintaining a vacuum of greater than 10 inches of mercury . a particularly preferred second vessel is a lee tri - mix turbo shear manufacturing vessel ( lee industries , newton , n . c .). the first step in the instant method is dissolving a polyalcohol and a permeation enhancer in an alkanol in an inert atmosphere . the alkanol is placed in a hemispherical , jacketed manufacturing vessel , an inert gas is used to blanket the alkanol in the vessel and the polyalcohol and the permeation enhancer are added to the alkanol while slowly stirring using a dispersion blade within the vessel . the polyalcohol is added to the alkanol in an amount between 1 % and 10 % by weight of the formulation and the permeation enhancer is added in an amount between 1 % and 30 % by weight of the formulation . the inert gas is preferably nitrogen . the alkanol may be a c 2 to c 4 alcohol , such as ethanol , isopropanol , or n - propanol , and is present in an amount between 35 % and 55 % by weight of the formulation . in certain preferred embodiments , the alkanol is ethanol . in other preferred embodiment , the ethanol is present in an amount of 47 . 5 %. in certain embodiments , the polyalcohol may be propylene glycol , butylene glycol , hexylene glycol , and ethylene glycol . in certain preferred embodiments , the polyalcohol is propylene glycol . the selection of the permeation enhancer can affect the amount and rate of transdermal absorption of hormone formulations . the amount of the permeation enhancer may be optimized . in one preferred embodiment , the permeation enhancer may comprise about 1 to 10 % of the formulation by weight . in one embodiment , the permeation enhancer may comprise about 5 % of the formulation by weight . in one embodiment , the permeation enhancer is a monoalkyl ether of diethylene glycol . the monoalkyl ether of diethylene is , for example , diethylene glycol monoethyl ether or diethylene glycol monomethyl ether . in certain preferred embodiments , the permeation enhancer is diethylene glycol monoethyl ether . in certain preferred embodiments , the permeation enhancer is diethylene glycol monoethyl ether in an amount of about 5 % by weight . the second step of the present methods is the addition of the therapeutically effective amount of the hormone to the stirred solution to form a testosterone solution . the hormone may advantageously be added as micronized particles and the solution is mixed between 450 and 550 rpm until fully dissolved . stirring usually occurs for 15 to 30 minutes to allow for the formation of a visually uniform solution lacking any visually detectable particles . a number of hormones are suitable for use in the methods disclosed herein . examples of estrogens for use herein include estrogen and estrogen derivatives such as 17 - beta - estradiol and esters thereof , ethinylestradiol , estriol ( trihydroxyestrin ), estrone , conjugated estrogens , in particular premarin , sodium estrone sulfate , 8 ( 9 )- dehydroestradiol derivatives , 17alfa - dihydroequilin , equilenin , 17alfa - dihydroequilenin , esterified estrogens , and equilin . when preparing transdermal estrogen formulations , the therapeutically effective amount of the estrogen in these transdermal formulations is between about 0 . 01 % and 1 . 00 % by weight of the formulation , preferably , between about 0 . 01 % and 0 . 1 % by weight of the formulation , and more preferably the therapeutically effective amount of the estrogen is about 0 . 06 % by weight of the formulation . in preparing the various progesterone formulations , the differences in the amount of progesterone added to the formulation can be balanced by the amount of purified water added on a w / w basis of the formulation . examples of progesterones for use herein include progesterone , norethisterone acetate , norgestrel , levonorgestrel , gestodene , cpa , chlormadinone acetate , drospirorenone , and 3 - ketodesogestrel . when preparing transdermal progesterone formulations , the progesterones are preferably in amount between : progesterone ( 10 %- 20 % by weight ); norethisterone acetate ( 1 . 5 %- 10 % by weight ); norgestrel ( 0 . 3 %- 0 . 8 % by weight ); levonorgestrel ( 0 . 12 % - 0 . 4 % by weight ); gestodene ( 0 . 08 %- 0 . 15 % by weight ); cpa ( 3 %- 7 % by weight ); chlormadinone acetate ( 3 %- 6 % by weight ); drospirorenone ( 3 %- 10 % by weight ); and / or 3 - ketodesogestrel ( 0 . 12 %- 0 . 5 % by weight ). in preparing the various progesterone formulations , the differences in the amount of progesterone added to the formulation can be balanced by the amount of purified water added on a w / w basis of the formulation . examples of testosterone compounds which may be used in the present invention include testosterone ( 17 - β - hydroxyandrostenone ), and testosterone esters , such as testosterone enanthate , testosterone propionate , testosterone decanoate and testosterone cypionate . the aforementioned testosterone esters are commercially available or may be readily prepared using techniques known to those skilled in the art or described in the pertinent literature . also , pharmaceutically acceptable esters of testosterone and 4 - dihydrotestosterone , typically esters formed from the hydroxyl group present at the c - 17 position ( such as enanthate , propionate , cypionate , phenylacetate , acetate , isobutyrate , buciclate , heptanoate , decanoate , undecanoate , caprate and isocaprate esters ); and pharmaceutically acceptable derivatives of testosterone such as methyl testosterone , testolactone , oxymetholone and fluoxymesterone may be used . in one embodiment , the therapeutically effective amount of the testosterone compound in the transdermal formulation produced by the methods herein is between about 0 . 50 % and 2 . 00 %, preferably , between about 0 . 75 % and 1 . 25 %, and more preferably the therapeutically effective amount of testosterone is about 1 . 00 %. [ by weight ?] the third step of the instant methods is the addition of purified water to form a hydroalcoholic mixture . preferably , the alkanol comprises about 45 % to 65 % and the water comprises about 35 % to 55 % of the hydroalcoholic mixture by weight . the hydroalcoholic mixture may be present in an amount of about 40 to 98 % by weight of the formulation , preferably between 80 % and 90 % by weight of the formulation . the fourth step of the instant methods is the addition of a sequestering agent . the sequestering agent is present from about 0 . 03 % to about 0 . 09 % w / w of the formulation depending on the type of compound . in preferred embodiments , the sequestering agent is edetic acid . a 3 % edetate disodium solution may be prepared , for instances , by dissolving 300 g of edetate disodium usp / ep in 10 kg purified water usp / ep by mixing at about 1 , 000 rpm for 25 min , and then added to the stirred hydroalcoholic solution to form a visually uniform second solution . the final step in the preparation of the primary compounding solution is the addition of a gelling agent to the second solution the gelling agent is added to the second solution in an amount sufficient to alter the viscosity of the formulation to result in the desired range of 16 , 000 to 40 , 000 cps . the gelling agent can be selected from the group including : carbomer , carboxyethylene or polyacrylic acid such as carbomer , carbopol 980 or 940 nf , 981 or 941 nf , 1382 or 1342 nf , 5984 or 934 nf , etd 2020 , 2050 , 934p nf , 971p nf , 974p nf , noveon aa - 1 usp ; cellulose derivatives such as ethylcellulose , hydroxypropylmethylcellulose ( hpmc ), ethylhydroxyethylcellulose ( ehec ), carboxymethylcellulose ( cmc ), hydroxypropylcellulose ( hpc ) ( klucel different grades ), hydroxyethylcellulose ( hec ) ( natrosol grades ), hpmcp 55 , methocel grades ; natural gums such as arabic , xanthan , guar gums , alginates ; polyvinylpyrrolidone derivatives such as kollidon grades ; and polyoxyethylene polyoxypropylene copolymers such as lutrol f grades 68 , 127 . other gelling agents may include chitosan , polyvinyl alcohols , pectins , and veegum grades . in certain preferred embodiments , the gelling agent is one of the carbomer , carbopol grade agents , preferably carbomer , carbopol 980 . the gelling agent may be present from about 0 . 5 % to about 10 . 0 % w / w depending on the type of polymer . in preferred embodiments , the gelling agent is carbomer , carbopol 980 present in an amount of 1 . 2 % by weight of the formulation . in certain embodiments , the primary compounding solution is transferred into a second round bottom , jacketed , pressure / vacuum vessel for ph adjustment , final mixing , deaeration , and gel formation . the primary compounding solution placed under vacuum , and mixed prior to the addition of the ph regulator . the ph regulator is generally a neutralizing agent , which can optionally have crosslinking functions . by way of example and not limitation , the ph regulator may include a ternary amine such as triethanolamine , tromethamine , tetrahydroxypropylethylendiamine , or may be a naoh solution . the ph regulator may be added in an amount between 0 . 05 % to about 2 % w / w . in preferred embodiments , the ph regulator is added in an amount between 0 . 2 % to about 0 . 5 % w / w . in certain preferred embodiments , the ph regulator is triethanolamine ( trolamine ). a 17 . 5 % trolamine solution may be readily prepared by dissolving 1750 g of trolamine nf in 10 kg purified water usp / ep , and the appropriate amount is calculated and added to the primary compounding solution to reach a final ph between 5 . 4 and 6 . 4 , which is readily determined by one skilled in the art . the ph regulator is added to the primary compound solution while stirring at low speeds , e . g ., 25 rpm , and then placed under vacuum , e . g ., 10 - 15 inches hg , for between 90 and 120 minutes to polymerize and deaerate the mixture to form a visually uniform gel yielding about 500 kg of the final hormone transdermal formulation . the final mixing step is conducted in a vacuum to prevent entrapped air during gelation and / or evaporation of the alkanol . the formulation may further include preservatives such as but not limited to benzalkonium chloride and derivatives , benzoic acid , benzyl alcohol and derivatives , bronopol , parabens , centrimide , chlorhexidine , cresol and derivatives , imidurea , phenol , phenoxyethanol , phenylethyl alcohol , phenylmercuric salts , thimerosal , sorbic acid and derivatives . the preservative may be present from about 0 . 01 to about 10 . 0 % w / w of the formulation depending on the type of compound . the formulation may further include buffers such as carbonate buffers , citrate buffers , phosphate buffers , acetate buffers , hydrochloric acid , lactic acid , tartric acid , diethylamine , triethylamine , triethanolamine , diisopropylamine , aminomethylamine . other buffers as known in the art may be included additionally or instead . the buffer may replace up to 100 % of the water amount within the formulation . optionally , the formulation may include moisturizers and / or emollients to soften and smooth the skin or to hold and retain moisture . by way of example and not limitation , moisturizers and emollients may include cholesterol , lecithin , light mineral oil , petrolatum , and urea . the formulation may further include anionic , non - ionic or cationic surfactants . the surfactant may be present from about 0 . 1 % to about 30 % w / w depending on the type of compound . the formulation may optionally include antioxidants such as but not limited to tocopherol and derivatives , ascorbic acid and derivatives , butylated hydroxyanisole , butylated hydroxytoluene , fumaric acid , malic acid , propyl gallate , metabisulfates and derivatives . the antioxidant may be present from about 0 . 001 to about 5 . 0 % w / w of the formulation depending on the type of compound . for any particular desired formulation , these other ingredients may be selected to achieve the desired drug delivery profile and the amount of penetration desired . the optimum ph may also be determined and may depend on , for example , the base and degree of flux required . the formulations produced by the methods of the present invention may be used in methods for treating a postmenopausal woman having hsdd . in one embodiment , the therapeutically effective amount of testosterone compound is applied directly to the skin the present transdermal therapy provides important advantages over the known oral , intramuscular , and transdermal products by advantageously delivering serum testosterone compound concentrations that are not subject to first - pass metabolism and avoiding wide swings in serum testosterone concentrations while reducing skin reactions often observed with existing transdermal patch products . the present formulation of the method may be applied once daily , or multiple times per day depending upon the condition of the woman . the formulation may be applied topically to any body part , such as the thigh , abdomen , shoulder , and upper arm . in one embodiment , a formulation in the form of a gel is applied to about a 5 inch by 5 inch area of skin application may be to alternate areas of the body as applications alternate . for example , the gel may be applied to the thigh for the first application , the upper arm for the second application , and back to the thigh for the third application . the amount of testosterone compound and dosing schedules necessary to provide a therapeutically effective amount may be monitored by following serum concentrations of testosterone . methods for measuring the serum levels of such hormones , particularly testosterone , are well known to one of ordinary skill in the art . the serum measures are preferably made when the therapeutically targeted level of steady state has been achieved . a metered dosage device for administration of the transdermal hormone formulation of the present invention may be used in connection with the methods herein . any metered dosage device capable of dispensing and administering the instant formulations of the methods may be used ( e . g ., see u . s . patent application publication no . us2006027064 ). preferably , the metered dosage device is capable of dispensing a predetermined , precise amount of a transdermal testosterone formulation produced by the present methods . for example , when used in combination with a gel containing 1 % testosterone , the metered dosage device may be designed to dispense 0 . 22 g of the topical formulation when activated e . g ., by pressing on the pump , such that about 2 mg of testosterone is dispensed . thus , the metered dosage device may be conveniently used for self - administration of a precise testosterone dosage . when smaller doses are to be applied more often , the device can be designed to dispense 1 . 0 or even 0 . 5 mg of testosterone upon each activation . thus , four 0 . 5 mg doses , two 1 mg doses or a single 2 mg dose can be administered to provide the preferred amount of 2 mg of testosterone each day . the following examples are illustrative and are not meant to be limiting . the following example describes the production of a 500 kg batch of a transdermal formulation according to the methods of the present invention . excluding preparation of certain reagents , all procedures throughout the manufacturing process were performed under a nitrogen blanket or vacuum and at ambient temperatures not exceeding 29 ° c . solutions were stirred at about 500 rpm unless otherwise noted . in a pre - charged , round bottom , jacketed , stainless steel 700 liter vessel , 223 kg of ethanol ( 200 proof ) usp / ep was stirred under high shear using a dispersion blade into which 30 kg of propylene glycol usp / ep and 25 kg diethylene glycol monoethyl ether ep / np were added . the mixture was stirred briefly until visually dissolved and then 173 kg of purified water usp / ep were added using a transfer pump , and the solution was further stirred . a 3 % edetate disodium solution was prepared by dissolving 300 g of edetate disodium usp / ep in 10 kg purified water usp / ep by mixing at about 1 , 000 rpm for 25 min , and then added to the stirred testosterone solution . after stirring for about 10 min , the speed of mixing was increased to about 650 rpm , and six kilograms of carbomer , carbopol 980 dispersion was slowly added to the mixture . the mixture was stirred for 75 minutes yielding a smooth , homogenous solution free of lumps ( the primary compounding solution ). the primary compounding solution was transferred into a round bottom , jacketed , stainless steel 1350 liter pressure / vacuum vessel for ph adjustment , final mixing , deaeration , and gel formation . the 700 liter vessel was rinsed with 15 kg of ethanol to ensure complete transfer , added to the primary compounding solution and placed under vacuum . a 17 . 5 % triethanolamine ( trolamine nf ) solution was prepared by dissolving 1750 g of trolamine nf in 10 kg purified water usp / ep . upon releasing the vacuum , a 7 . 64 kg amount of trolamine solution was added to primary compounding solution while stirring at 10 rpm . the mixture was stirred at 25 rpm under vacuum ( 11 inches of hg ) for 101 minutes to polymerize and deaerate the mixture forming a visually uniform gel yielding about 500 kg of the final transdermal formulation . the final transdermal formulation consisted of : 47 . 5 % ethanol ; 6 % propylene glycol ; 5 % diethylene glycol monoethyl ether ; 1 . 2 % carbomer , carbopol 980 ; 0 . 35 % triethanolamine ; 0 . 06 % edetate disodium ; and 38 . 89 % water . the viscosity of the transdermal gel formulation was 22 , 000 - 25 , 000 cps ( range 16 , 000 - 40 , 000 ) having a slightly acidic ph , between 5 and 7 . the following example describes the production of a 500 kg batch of a 1 % testosterone transdermal formulation . excluding the preparation of certain reagents , all procedures throughout the manufacturing process were performed under a nitrogen blanket or vacuum and at ambient temperatures not exceeding 29 ° c . solutions were stirred at about 500 rpm unless otherwise noted . in a pre - charged , round bottom , jacketed , stainless steel 700 liter vessel , 223 kg of ethanol ( 200 proof ) usp / ep was stirred under high shear using a dispersion blade into which 30 kg of propylene glycol usp / ep and 25 kg diethylene glycol monoethyl ether ep / np were added . the mixture was stirred briefly until visually dissolved and then five kilograms of testosterone micronized usp / ep were added . after stirring for 17 min , 168 kg of purified water usp / ep were added using a transfer pump , and the solution was further stirred . a 3 % edentate disodium solution was prepared by dissolving 300 g of edetate disodium usp / ep in 10 kg purified water usp / ep by mixing at about 1 , 000 rpm for 25 min , and then added to the stirred testosterone solution . after stirring for about 10 min , the speed of mixing was increased to about 650 rpm , and six kilograms of carbomer , carbopol 980 dispersion was slowly added to the mixture . the mixture was stirred for 75 minutes yielding a smooth , homogenous solution free of lumps ( the primary compounding solution ). the primary compounding solution was transferred into a round bottom , jacketed , stainless steel 1350 liter pressure / vacuum vessel for ph adjustment , final mixing , deaeration , and gel formation . the 700 liter vessel was rinsed with 15 kg of ethanol to ensure complete transfer , added to the primary compounding solution and placed under vacuum . a 17 . 5 % triethanolamine ( trolamine nf ) solution was prepared by dissolving 1750 g of trolamine nf in 10 kg purified water usp / ep . upon releasing the vacuum , a 7 . 64 kg amount of trolamine solution was added to primary compounding solution while stirring at 10 rpm . the mixture was stirred at 25 rpm under vacuum ( 11 inches of hg ) for 101 minutes to polymerize and deaerate the mixture forming a visually uniform gel yielding about 500 kg of the final testosterone transdermal formulation . the final transdermal testosterone formulation consisted of : 1 % testosterone ; 47 . 5 % ethanol ; 6 % propylene glycol ; 5 % diethylene glycol monoethyl ether ; 1 . 2 % carbomer , carbopol 980 ; 0 . 35 % triethanolamine ; 0 . 06 % edetate disodium ; and 38 . 89 % water . the viscosity of the transdermal gel formulation was 22 , 000 - 25 , 000 cps ( range 16 , 000 - 40 , 000 ) having a slightly acidic ph , between 5 and 7 .

methods for commercial production of transdermal formulations comprising a hormone compound are provided . in particular , methods for commercial scale production under an inert atmosphere of a transdermal formulation comprising a therapeutically effective amount of a hormone , preferably a testosterone compound , useful for the treatment of hypoactive sexual desire disorder in postmenopausal women are provided .

the choice and configuration of the fabric can have a significant impact on the effectiveness of the pack in use . important parameters of the fabric include insulation qualities and heat distribution , moisture control and comfort . with these parameters in mind , the inventors have found that a combination of at least two layers of fabric , each having a different function , can simultaneously provide moisture control and heat distribution over a wide temperature range . this result was unexpected , since it was heretofore believed . that moisture control and heat distribution were mutually exclusive in a single wrap . in a preferred embodiment , suitable yarn for temperature control has thermal properties which vary as a function of temperature . standardized measurement of thermal properties of yarn under static and dynamic temperature conditions is provided in accordance with test method for steady state and dynamic thermal performance in textile materials ( astm d7024 ). in a highly preferred embodiment , the temperature control yarn incorporates one or more phase change materials which act as temperature buffers . such materials undergo a phase change as the surrounding temperature varies , absorbing heat during a temperature rise , and releasing heat when temperature falls . the melting point of the phase change materials , that is , the temperature at which the phase change occurs , can be varied depending on the particular application . a source for suitable fabrics incorporating phase change materials is outlast technologies of boulder , colo . in addition to the selection of particular phase change materials , the construction of the temperature control yarn can be varied by the use of specific denier and loop size in the terry . moisture control can be accomplished by the use of a wicking yarn effectively moving the moisture to enhance effectiveness and safety . various moisture wicking yarns are available , one of which is sorbtek manufactured by unifi . fig1 illustrates one embodiment of the invention comprising a sleeve 1 in which a thermal pack 2 is encased within a moisture absorbing fabric 3 . a temperature control fabric 4 is positioned adjacent the thermal pack along one side thereof . in an alternative embodiment , an insulation fabric can be positioned adjacent the pack on a side opposite the temperature control fabric 4 . the sleeve 1 is designed to be placed directly over the affected area , for example , a knee . the sleeve is oriented with the temperature control fabric positioned between the affected area and the thermal pack 2 . the optional insulation fabric reduces heat loss from the thermal pack from the side opposite to that applied to the affected area . fig2 illustrates another embodiment of the invention comprising a wrap 10 . the wrap 10 includes a thermal pack 11 and a temperature control fabric 12 . the pack 11 and fabric 12 are contained in a sleeve 13 comprising a moisture absorbing fabric 13 . in an alternative embodiment ( not shown ), a sleeve of suitable fabric can be knitted on a circular knitting machine and a moisture absorbing fabric is plated to the outside of the sleeve and a temperature control fabric is constructed with a terry knit to the inside of the sleeve . the moisture absorbing material can also knitted around the entire circumference with the terry covering half the circumference . additional yarns such as various well know acrylics may be added to manipulate the fabric weight . the wrap can be constructed using a flat knit machine or weaving . the two layers of the moisture absorbing fabric and temperature control fabric can be woven using a jacquard machine or the fabrics can be woven separately and then sewn together . a thermocromatic ink can be printed on the fabric to indicate to the user the temperature of the pack when it is heated such as in a microwave or cooled such as a freezer . this would indicate that the pack has reached the proper temperature , that it is too hot or that it is sufficiently cold .

an article for applying hot or cold therapy to an individual in need of such therapy which includes a sleeve containing a thermal pack encased within a moisture absorbing fabric and a temperature control fabric adjacent the thermal pack .

referring to fig1 - 3 , the surface of a wall member 10 according to the present invention may be substantially divided into several equal squares . a doorcase 11 with an openable door and a window frame 12 with an openable window are provided on the wall member 10 . each of several properly arranged squares of the wall member is provided with a circular hole 13 of which the center coincides with the center of its corresponding square . provided on each of the rear surfaces of certain properly arranged squares of the wall member 10 is a cylindrical protrusion 14 with its axis passing through the center of its corresponding square . the wall member 10 is preferably formed by injection - moulding . in order to make the wall member capable of being bent to form various angles and into various shapes , several lines shared by the squares are provided with several longitudinal and transverse grooves 15a , 15b and 15c . it is to be noted that the surface of the wall member 10 can be vivified by stripes , colors and patterns , such as a pattern of bricks . referring to fig4 - 6 , the surface of a roof member 20 according to the present invention may be substantially divided into several equal squares . each of several properly arranged squares of the roof member 20 is provided with a cylindrical recess 21 of which the center coincides with the center of its corresponding square . each of the rear surfaces of certain of said squares is provided with a cylindrical protrusion 22 with its axis passing through a certain point on a transverse bisecting line of its corresponding square . in order to combine two roof members 20 together , each edge portion of said certain lateral squares thereof is provided with a semicircular protrusion 23 and a homocentric flange 24 surrounding said protrusion 23 . there is a certain semiannular clearance 25 between said flange 24 and said protrusion 23 . the center of the semicircular protrusion 23 is situated on said transverse bisecting line of its corresponding square on which is also located said cylindrical projection 22 . in order to enable the roof member 20 , which is preferably formed by injection - moulding , to be bent freely to form various angles , along lines shared by the squares thereof , a plurality of longitudinal and / or transverse grooves 26 are located along these lines . referring to fig7 a roof connecting member 30 has two parallel grooves 31 , 32 thereon for receiving the cylindrical protrusion 22 , a cylindrical protrusion on the underside thereof and a longitudinal groove 34 on a lateral surface thereof . the width of each of said two parallel grooves 31 , 32 is equal or slightly smaller than the outer diameter of the plug 22 of the roof member 20 so that the protrusion 22 may not only be received in any of the grooves 31 , 32 , but may also be glided and fixed to the desired place . referring to fig8 a roof support member 40 which is mainly triangular or trapezoidal has two protrusions 41 and 42 provided on the rear surface thereof for being inserted into the longitudinal groove 34 of said roof connecting member 30 . likewise , the outer diameter of said protrusion 41 or 42 is equal or slightly larger than the width of the longitudinal groove of the roof connecting member 30 , so that the protrusions 41 , 42 may not only be received in , but may also glide and be fixed to the longitudinal groove 34 . an edge portion of the roof member 20 can be supported by and abutted against the lateral rims of the roof support member 40 . referring to fig9 each center of five sides of a cubic connecting member 50 is provided with a cylindrical recess 51 , and the center of the remaining one side thereof is provided with a cylindrical protrusion 52 . the inner diameter of the cylindrical recess 51 is equal to or slightly smaller than the outer diameter of the cylindrical protrusion 52 , of the cylindrical protrusion 33 of the roof connecting member 30 and of the cylindrical protrusion 14 as well as the diameter of the circular hole 13 of the wall member 10 , so that said cylindrical recess 51 may not only receive the protrusion 52 of another cubic connecting member 50 to form a jamb , but also connect with the roof connecting member 30 and the wall member 10 . furthermore , the area of each side of the cubic connecting member 50 is equal to that of each equal square of the wall member 10 and the roof member 20 . the structure of a base board 60 as shown in fig1 is substantially the same as that depicted in my u . s . pat . no . 4 , 164 , 091 . the base board has a plurality of cylindrical recesses 61 for receiving tightly the protrusion 52 of the cubic connecting member 50 and serves as the base of an assembled house . the hollow cylindrical connector 70 as shown in fig1 is in fact also disclosed in the u . s . pat . no . 4 , 164 , 091 . the cylindrical connector 70 is mainly used for connecting the base boards 60 as well as for being inserted into an annular cleance formed by pairing two semicircular protrusions 23 and homocentric flanges 24 provided respectively on two roof members 20 . moreover , by inserting the connector 70 through the circular hole 13 of the wall member 10 and into the cylindrical recess 51 of a jamb or a beam formed by the cubic connecting members 50 , other members can be further connected thereto so as to diversify shapes of the assembled houses . diversified configurations of houses e . g . a shape as shown in fig1 can be made by various connections of the above - mentioned main members . in the operation of constructing such a house , several of said base boards 60 are firstly combined with one another to form a large base ground . then several jambs , formed by connecting cubic connecting members 50 , are positioned on the ground . thereafter , the wall members 10 are arranged to surround and connect the jambs by engaging the protrusions 14 provided on the rear surfaces of the wall members with the cylindrical recess 51 of the jambs . because the wall member 10 has more than one bendable groove 15a , 15b and 15c thereon , it can act as a more than one side wall ; secondly , the cylindrical protrusion 33 provided on the underside of the roof connecting member 30 is inserted into the cylindrical recess 51 on the top side of the jamb , while two longitudinal recesses 34 of two said roof connecting members 30 are engaged respectively with two cylindrical protrusions 41 , 42 of the roof support member 40 ; finally , the roof member 20 is folded according to the angle of the roof support member 40 and supported by inserting the protrusion 22 provided on the rear surface of the roof member into two parallel grooves 31 , 32 of the roof connecting member , while the lateral edge portion of the roof member 20 is placed on the roof support member 40 . it is to be noted that a variety of elements other than the aforementioned members might be added to vivify the outer appearance of the assembled house , e . g . a parapet 91 and a pergola pillar 90 as shown in fig1 which are not depicted in the above . such kind of obvious modifications and variations of the present invention should fall within the spirit and the scope of the present invention . it is hence intended that the scope of the present invention be defined by the appended claims .

a set of building blocks readily assembled to form a variety of houses comprises several flexible wall elements , flexible roof elements , roof forming elements , roof connecting elements , cubic elements , and base boards which can be connected side by side to construct a base as a ground member of the houses . the wall elements can be bent into different configurations so as to act as side walls at one &# 39 ; s desire .

a first aspect of the invention comprises a method for preventing or treating hiv - 1 infection which comprises administering , to a human who has been exposed to or infected by hiv - 1 , a prophylactically or therapeutically effective amount of a dibenz [ d , f ][ 1 , 4 ] oxazepin ( or thiazepin )- 11 ( 10h )- one or - thione of the formula i ## str1 ## wherein , x is oxygen or sulfur ; r 1 is hydrogen , alkyl of 1 to 6 carbon atoms , fluoroalkylmethyl of 1 to 3 fluorine atoms and 2 to 6 carbon atoms , alkenyl or alkynyl of 2 to 6 carbon atoms , mono - or dihalovinyl , cycloalkyl of 3 to 6 carbon atoms , alkyloxyalkyl or alkylthioalkyl of 2 to 6 carbon atoms , alkanoyl of 2 to 4 carbon atoms , arylmethyl or arylmethyloxy or arylcarbonyl ( wherein the aryl moiety is phenyl , thienyl or furanyl optionally substituted with methyl , methoxy or halogen ), alkoxycarbonylalkyl of 3 to 6 carbon atoms , aminoalkyl of 1 to 3 carbon atoms , mono - or di - alkylaminoalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , alkanoylamtnoalkyl wherein the alkanoyl moiety contains 2 to 3 carbon atoms and the alkyl moiety contains 1 to 2 carbon atoms , amtnocarbonylalkyl of 2 to 4 carbon atoms , mono - or dfalkylaminocarbonylalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms or hydroxyalkylmethyl of 2 to 6 carbon atoms ; r 3 is hydrogen , alkyl of 1 to 4 carbon atoms , halogen , hydroxyl , alkoxy of 1 to 3 carbon atoms , alkylthio of 1 to 3 carbon atoms , alkanoyloxy of 2 to 3 carbon atoms , amino , alkylamino of 1 to 2 carbon atoms , aminoalkyl of 1 to 2 carbon atoms , mono - or dimethylaminomethyl , hydroxyalkyl of 1 to 4 carbon atoms , alkoxyalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , alkylthioalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , carboxylalkyl of 2 to 4 carbon atoms , carboxyalkoxy of 2 to 3 carbon atoms , alkoxycarbonylmethyl of 3 to 4 carbon atoms or methoxycarbonylmethoxy ; r 5 is hydrogen , alkyl of 1 to 4 carbon atoms , halogen , hydroxyl , alkoxy of 1 to 3 carbon atoms , alkylthio of 1 to 3 carbon atoms , alkanoyloxy of 2 to 3 carbon atoms , amino or alkylamino of 1 to 2 carbon atoms ( excepting 4 - amino and 4 - alkylamino ), aminoalkyl of 1 to 2 carbon atoms , mono - or dimethylaminomethyl , hydroxyalkyl of 1 to 4 carbon atoms , alkoxyalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , carboxylalkyl of 2 to 4 carbon atoms , carboxyalkoxy of 2 to 3 carbon atoms , alkoxycarbonylmethyl of 3 to 4 carbon atoms or methoxycarbonylmethoxy ; in a subgeneric aspect , the invention comprises the above - described method wherein , in the compound of formula i , r 1 is alkyl of 1 to 5 carbon atoms , fluoroalkylmethyl of 1 to 3 fluorine atoms and 2 to 5 carbon atoms , alkenyl or alkynyl of 2 to 5 carbon atoms , mono - or dihalovinyl , cycloalkyl of 3 to 6 carbon atoms , alkyloxymethyl or alkylthiomethyl of 2 to 5 carbon atoms , alkoxyethyl or alkylthioethyl of 3 to 5 carbon atoms , alkanoyl of 2 to 3 carbon atoms , arylmethyl ( wherein the aryl moiety is phenyl , thienyl or furanyl , optionally substituted with methyl , methoxy or halogen ), alkoxycarbonylmethyl of 3 to 5 carbon atoms , acetylaminoalkyl wherein the alkyl moiety contains 1 to 2 carbon atoms , alkoxy of 1 to 4 carbon atom or hydroxyalkylmethyl of 2 to 5 carbon atoms ; r 3 is hydrogen , alkyl of 1 to 3 carbon atoms , halogen , hydroxyl , methoxy , ethoxy , methylthio , ethylthio , alkanoyloxy of 2 to 3 carbon atoms , amino , methylamino , aminoalkyl of 1 to 2 carbon atoms , mono - or dimethylaminomethyl , hydroxyalkyl of 1 to 3 carbon atoms , alkoxyalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , alkylthioalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , carboxylalkyl of 2 to 3 carbon atoms , carboxyalkoxy of 2 to 3 carbon atoms , alkoxycarbonylmethyl of 3 to 4 carbon atoms or methoxycarbonylmethoxy ; r 5 is hydrogen , alkyl of 1 to 3 carbon atoms , halogen , hydroxyl , methoxy , ethoxy , methylthio , ethylthio , alkanoyloxy of 2 to 3 carbon atoms , amino or methylamino ( excepting 4 - amino and 4 - methylamino ), aminoalkyl of 1 to 2 carbon atoms , alkoxyalkyl or alkylthioalkyl wherein each alkyl moiety contains 1 to 2 carbon atoms , carboxyalkoxy of 2 to 3 carbon atoms , carboxyalkoxy of 2 to 3 carbon atoms , alkoxycarbonylmethyl of 3 to 4 carbon atom or methoxycarbonylmethoxy . in a further subgeneric aspect , the invention comprises the above described method wherein , in the compound of formula i , r 1 is alkyl of 1 to 4 carbon atoms , fluoroalkylmethyl of 1 to 3 fluorine atoms and 2 to 4 carbon atoms , alkenylmethyl or alkynylmethyl of 2 to 4 carbon atoms , mono - or 1 , 2 - dihalovinyl , alkoxymethyl or alkylthiomethyl of 2 to 4 carbon atoms , alkoxyethyl or alkylthioethyl of 3 to 4 carbon atoms or alkoxycarbonylmethyl of 3 to 4 carbon atoms ; r 3 hydrogen , methyl , ethyl , chlorine , bromine , hydroxyl , methoxy , methylthio , acetyloxy , amino , methylamino , aminomethyl , hydroxymethyl , hydroxyethyl , methoxymethyl or methylthiomethyl ; r 5 is hydrogen , methyl , ethyl , chlorine , bromine , hydroxyl , methoxy , methylthio , acetyloxy , amino or methylamino ( excepting 4 - amino and 4 - methylamino ), aminomethyl , hydroxymethyl , hydroxyethyl , methoxymethyl or methylthiomethyl . in a still further subgeneric aspect , the invention comprises the above described method wherein , in the compound of formula i , r 1 is alkyl of 1 to 4 carbon atoms , fluoroalkylmethyl of 1 to 3 fluorine atoms and 2 to 4 carbon atoms , alkenylmethyl or alkynylmethyl of 2 to 4 carbon atoms , mono - or 1 , 2 - dihalovinyl , alkoxymethyl or alkylthiomethyl of 2 to 4 carbon atoms , alkoxyethyl or alkylthioethyl of 3 to 4 carbon atoms or alkoxycarbonylmethyl of 3 to 4 carbon atoms ; compounds of formula i are either known or can be prepared using methods analogous to those used to prepare known compounds . representative compounds of formula i and methods for their preparation are described , for example , in the following prior art references : u . s . pat . nos . 3 , 367 , 930 ; 3 , 541 , 085 ; 3 , 546 , 214 ; and , 4 , 379 , 150 ; and british patent nos . 1 , 164 , 579 and 1 , 170 , 322 . compounds of formula i , can additionally be prepared according to the following general methods a , b and c . compounds of formula i , wherein z is oxygen and x and r 1 - r 5 have the meanings given above with the exception of r 1 being hydrogen , may be obtained , for example , by converting a compound of the formula ii ## str2 ## wherein r 2 - r 5 are as defined above , into the corresponding alkali or alkaline earth methal compounds of the formula iii ## str3 ## wherein r 2 - r 5 are as defined above , and subsequently reacting , without isolation , this alkali metal compound with a reactive alkylating or acylating reagent of the formula iv wherein r 1 has the meanings given above except for hydrogen and y is a suitable leaving group such as chloride , bromide , iodide , an alkyl or arylsulfonate , or an alkyl - or arylcarbonyloxy group under well known alkylating or acylating conditions . it will be obvious to those skilled in the art that the presence of nucleophilic substituents in compounds of formula ii , for example , will require the use of an intermediate having substituents which are , other than the 5 - position nitrogen , not nucleophilic but which can be derivatized to yield the required group . for example , amino or monoalkylamino substituents are preferably obtained by alkylating or acylating an intermediate of formula ii having nitro group ( s ) at the desired positions , and subsequently reducing the nitro group ( s ), and alkylating , if appropriate , to yield the final product . compounds of formula i wherein z is oxygen and x and r 1 - r 5 are as defined above may be obtained by cyclization of compounds of the formula v ## str4 ## wherein x and r 2 - r 5 are as defined above and hal is fluorine , chlorine , bromine or iodine , preferably in the presence of an inorganic base , such as sodium or potassium hydride , lithium alkyls such as n - butyl lithium , sodium or potassium hydroxide , or in the presence of an organic base such as quinoline or 4 -( n , n - dimethylamino ) pyridine , at ambient or elevated temperatures , preferably 80 °- 175 ° c ., up to the boiling point of the reaction mixture . in those cases where r 1 is hydrogen , 2 equivalents of base should be used . suitable solvents include inert aprotic solvents such as sulfolane or dimethylformamide . the diphenylamides of formula v may be obtained , for example , by condensing suitably substituted ortho halobenzoic acid chlorides of the formula vi ## str5 ## wherein hal may be fluorine , chlorine , bromine or iodine and r 4 and r 5 are as defined above , with ortho - amino - phenols ( or thiophenols ) of the formula vii ## str6 ## wherein x , r 2 and r 3 are as defined above , under well - known reaction conditions . depending upon the reaction conditions employed and the nature of x and r 2 - r 5 , tricyclic compounds of the formula ii , may be formed in one step , without the isolation of the amide of formula v , by the condensation of compounds of the formulas vi and vii . this single - step formation of the tricyclic compounds is most readily effected when x is sulfur and at elevated temperatures , especially in the range of 125 °- 200 ° c . thiolactams of the general formula i , wherein x and r 1 - r 5 are as defined above , can be obtained by treatment of lactams of the formula i with sulfurating reagents such as 2 , 4 - bis ( 4 - methoxyphenyl )- 1 , 3 - dithia - 2 - 4 - diphosphetane - 2 , 4 - disulfide , bis ( tricyclohexyltin ) sulfide , bis ( tri - n - butyltin ) sulfide , bis ( triphenyltin ) sulfide , bis ( trtmethylsilyl ) sulfide and phosphorous pentasulfide . the reaction is generally carried out under anhydrous conditions in inert organic solvents such as carbon disulfide , benzene or toluene , at room temperature or , preferably , at higher temperatures up to the boiling point of the reaction mixture . when using the above mentioned tin or silyl sulfides it is preferable to carry out the sulfurization reaction in the presence of a lewis acid such as boron trichloride . it will be obvious to those skilled in the art that the presence of another carbonyl moiety in a compound of formula i , for example , a compound wherein z is oxygen and any of r 2 to r 5 is alkanoyl , will require that the ketone carbonyl be protected via known methods by a suitable protecting group ( such as ethylene glycol ) prior to the sulfurization reaction ; deprotection subsequent to the sulfurization reaction provides the desired compound . similarly , in cases wherein r 1 is , for example , acetyl , it will be obvious that the sulfurization reaction should be performed prior to the acylation ( of n - 5 ). in those cases wherein the substituents at r 2 to r 5 can be derived from nitro , for example , alkanoylamino , the sulfurization reaction can be performed on the corresponding nitro derivative , followed by an appropriate ( known ) reduction and finally acylation to yield the desired product . compounds of formula i having basic or acidic substituents may , if desired , be converted into their pharmaceutically acceptable salts by conventional methods . examples of inorganic and organic acids which may form pharmaceutically acceptable acid addition salts with a compound of formula i having basic substituents are the following : hydrochloric acid , hydrobromic acid , sulfuric acid , phosphoric acid , nitric acid , tartaric acid , citric acid , methanesulfonic acid and the like . examples of bases which may form pharmaceutically acceptable salts with compounds of formula i having acidic substituents are the following : sodium hydroxide , potassium hydroxide , calcium hydroxide , ammonia , tromethamine and the like . the above described compounds of formula i inhibit hiv - 1 reverse transcriptase and thereby inhibit hiv - 1 replication , making them useful in the method which constitutes one aspect of the invention . in carrying out this method , the compounds of formula i may be administered in single or divided doses by the oral , parenteral or topical routes . a suitable oral dosage for such compounds would be in the range of about 10 to 500 mg per day . in parenteral formulations , a suitable dosage unit may contain from 1 to 50 mg of said compounds , whereas for topical administration , formulations containing 0 . 01 to 1 % active ingredient are preferred . it should be understood , however , that the dosage administration will vary from patient to patient and the dosage for any particular patient will depend upon the clinician &# 39 ; s judgement , who will use as criteria for fixing a proper dosage the size and condition of the patient as well as the patient &# 39 ; s response to the drug . when such compounds are to be administered by the oral route , they may be administered as medicaments in the form of pharmaceutical preparations which contain them in association with a compatible pharmaceutical carrier material . such carrier material can be an inert organic or inorganic carrier material suitable for oral administration . examples of such carrier materials are water , gelatin , talc , starch , magnesium stearate , gum arabic , vegetable oils , polyalkyleneglycols , petroleum jelly and the like . the pharmaceutical preparations can be prepared in a conventional manner and finished dosage forms can be solid dosage forms , for example , tablets , dragees , capsules , and the like , or liquid dosage forms , for example , solutions , suspensions , emulsions and the like . the pharmaceutical preparations may be subjected to conventional pharmaceutical operations such as sterilization . further , the pharmaceutical preparations may contain conventional adjuvants such as preservatives , stabilizers , emulsifiers , flavor - improvers , wetting agents , buffers , salts for varying the osmotic pressure and the like . solid carrier material which can be used include , for example , starch , lactose , mannitol , methyl cellulose , microcrystalline cellulose , talc , silica , dibasic calcium phosphate , and high molecular weight polymers , such as polyethylene glycol . for parenteral use , it is preferred to administer such compounds in an aqueous or non - aqueous solution , suspension or emulsion in a pharmaceutically acceptable oil or a mixture of liquids , which may contain bacteriostatic agents , antioxidants , preservatives , buffers or other solutes to render the solution isotonic with the blood , thickening agents , suspending agents or other pharmaceutically acceptable additives . additives of this type include , for example , tartrate , citrate and acetate buffers , ethanol , propylene glycol , polyethylene glycol , complex formers ( such as edta ), antioxidants ( such as sodium bisulfite , sodium metabisulfite , and ascorbic acid ), high molecular weight polymers ( such as liquid polyethylene oxides ) for viscosity regulation and polyethylene derivatives of sorbitol anhydrides . preservatives may also be added if necessary , such as benzoic acid , methyl or propyl paraben , benzalkonium chloride and other quaternary ammonium compounds . the compounds can also be administered as solutions for nasal applications which may contain , in addition to the compounds , suitable buffers , tonicity adjusters , microbial preservatives , antioxidants and viscosity - increasing agents in an aqueous vehicle . examples of agents used to increase viscosity are polyvinyl alcohol , cellulose derivatives , polyvinylpyrrolidone , polysorbates or glycerin . microbial preservatives added may include benzalkonium chloride , thimerosal , chlorobutanol or phenylethyl alcohol . as stated before , the compounds of formula i inhibit the enzymatic activity of hiv - 1 rt . based upon testing of these compounds , as described below , it is known that they inhibit the rna - dependent dna polymerase activity of hiv rt . it is believed that they also inhibit the dna - dependent dna polymerase activity of hiv rt . utilizing the reverse transcriptase ( rt ) assay described below , compounds can be tested for their ability to inhibit the rna - dependent dna polymerase activity of hiv rt . certain specific compounds , described in the examples which appear below , were so tested . the results of this testing appear in table i , below . among the enzymes for which human immunodeficiency virus ( hiv - 1 ) encodes is a reverse transcriptase ( 1 ), so - named because it transcribes a dna copy from an rna template . this activity can be quantitatively measured in a cell - free enzyme assay which has been previously described ( 2 ), and is based upon the observation that reverse transcriptase is able to use a synthetic template [ poly r ( c ) primed with oligo d ( g )] to transcribe a radio - labelled , acid - precipitable dna strand utilizing 3 h - dgtp as a substrate . reverse transcriptase enzyme from the lav strain of human immuno - deficiency virus ( hiv - 1 ) ( 1 ) was isolated from the bacterial strain jm109 ( 3 ) expressing the dna clone pbrtprtl +( 2 ) which is under the control of the lac promotor in the expression vector pibi21 ( 4 ). an overnight culture grown in 2xyt medium ( 37 ° c ., 225 rpm ) ( 5 ) supplemented with 100 mg / ml ampicillin for positive selection is inoculated at a 1 : 40 dilution into m9 medium supplemented with 10 mg / ml thiamine , 0 . 5 % casamino acids , and 50 mg / ml ampicillin ( 5 ). the culture is incubated ( 37 ° c ., 225 rpm ) until it reaches an od540 of 0 . 3 - 0 . 4 . at that time the repressor inhibitor iptg ( isopropyl b - d - thiogalactopyranoside ) is added to 0 . 5 mm and incubated for 2 additional hours . bacteria are pelletted , resuspended in a 50 mm tris , 0 . 6 mm edta , 0 . 375m nacl buffer and digested by the addition of lysozyme ( 1 mg / ml ) for 30 minutes on ice . the cells are lysed by the addition to 0 . 2 % np - 40 and brought to 1m nacl . after removal of the insoluble debris by centrifugation , the protein is precipitated by the addition of 3 volumes of saturated aqueous ammonium sulfate . the enzyme is pelleted , resuspended in rt buffer ( 50 mm tris ph 7 . 5 , 1 mm edta , 5 mmdtt , 0 . 1 % np - 40 , 0 . 1m nacl , and 50 % glycerol ) and stored at - 70 ° c . for further use . ______________________________________stock reagent 2x mix concentration______________________________________1m tris ph 7 . 4 100 mm1m dithiothrietol 40 mm1m nacl 120 mm1 % nonidet p - 40 0 . 1 % 1m mgcl 4 mm [ poly r ( c )/ oligo d ( g )]( 5 : 1 ) 2 mg / ml . sup . 3 h - dgtp ( 81 μm ) 0 . 6 mm______________________________________ the 2x concentrated stock reaction mixture is aliquoted and stored at - 20 ° c . the mixture is stable and thawed for use in each assay . this enzyme assay has been adapted to a 96 well microtiter plate system , and has been previously described ( 6 ). tris buffer ( 50 mm , ph 7 . 4 ), vehicle ( solvent diluted to match the compound dilution ), or compounds in vehicle are dispensed into 96 - well microtiter plates ( 10 μl / well ; 3 wells / compound ). the hiv rt enzyme is thawed , diluted in 50 mm tris ph 7 . 4 so that fifteen μl of diluted enzyme contain 0 . 001 unit ( one unit is that amount of enzyme to transform 1 micromole of substrate per minute at 25 ° c . ), and 15 μl are dispensed per well . twenty μl of 0 . 12 - 0 . 5m edta are added to the first three wells of the microtiter plate . edta chelates the mg ++ present and prevents reverse transcription . this group serves as background polymerization which is subtracted from all other groups . twenty - five μl of the 2x reaction mixture are added to all wells and the assay is allowed to incubate at room temperature for 60 minutes . the assay is terminated by precipitating the dna in each well with 50μl of 10 % trichloracetic acid ( tca ) in 1 % sodium pyrophosphate . the microtiter plate is incubated for 15 minutes at 4 ° c . and the precipitate is fixed onto # 30 glass fiber paper ( schleicher & amp ; schuell ) using a skatron semi - automatic harvester . the filters are then washed with additional 5 % tca containing 1 % sodium pyrophosphate , rinsed with 70 % aqueous ethanol , dried , and transferred to scintillation vials ( 6 ). each vial receives 2 mls of scintillation cocktail and is counted in a beckman beta counter . 3 . yanisch - perron , c ., viera , j ., and messing , j ., 3 ., gene 33 : 103 , 1985 5 . maniatis , t , fritsch , e . f ., and j . sambrook , eds . molecular cloning : a laboratory manual , cold spring harbor laboratory , 1982 in order to confirm that compounds which are active in the rt assay also have the ability to inhibit hiv replication in a living system , a compound of formula i was also tested in the human t - cell culture assay described below . the results of this test appears in table ii . assay theory : formation of syncytia is a feature of in vitro cultures of cd4 + t - cells infected with hiv - 1 . in this assay , t - cells are treated with a putative replication inhibiting compound and then infected with hiv - 1 . after incubation the culture is checked for the formation of syncytia . the absence or reduction is the number of syncytia is used as a measure of the test compound &# 39 ; s ability to inhibit hiv replication . assay method : the target cells , designated c8166 , are a subclone of human lymphoma cells of t - cell origin and are established at an initial density of 5 × 10 4 per 100 μl in rpmi 1640 (+ 10 % fetal bovine serum ) culture medium in 96 well flat bottom plates . a selected amount of test compound , dissolved in dmso is included . after 24 hours , 50 - 100 tcid 50 &# 39 ; s ( the dose that results in induced effect in 50 % of test cultures ) of the htlv - iiib strain of hiv - 1 ( 2 ) are innoculated into each culture . control cultures receive compound or virus only . four days after virus challenge , cultures are visually examined for the frequency and distribution of virus - induced giant cell syncytia . the percent inhibition by the test compound is determined by comparison with control values . confirmation of the presence or absence of virus replication is accomplished by harvesting the cell free culture fluids from all experimental groups to determine the presence or absence of infectious progeny through the induction of syncytia formation in secondary human t - cell cultures after 3 days . ( 1 ) m . somasundaran and h . l . robinson , science 242 , 1554 ( 1998 ) ( 2 ) g . m . shaw , r . h . hahn , s . k . arya , j . e . groopman , r . c . gallo and f . wong - staal , science 226 , 1165 ( 1984 ) in order to assess the specificity of the enzyme inhibitory activity of the compounds of formula i , a few were tested , using known per se assay methods , for their ability to inhibit feline leukemia virus - derived reverse transcriptase and calf thymus - derived dna alpha - polymerase . none of the compounds so tested was observed to possess any inhibitory activity against these enzymes . these results indicate that the enzyme inhibitory activity of the compounds provided by the invention is directed rather specifically against hiv rt . in order to roughly assess the cytotoxicity of the compounds of formula i , two such compounds were tested in the mtt cellular cytotoxicity assay described below . the results of this testing are reported in table ii , below . compounds having a relatively high ec 50 are preferred . the mtt ( 3 -( 4 , 5 - dimethylthiazol - 2yl )- 2 , 5 diphenyl tetrazolium bromide ) assay is based on cleavage of tetrazolium bromide by metabolically active cells , resulting in a highly quantitative blue color . this assay has been previously described ( 1 ) but has been optimized for the purposes of the testing reported herein . the h9 cell line ( 2 ), an established human lymphoma suspension cell line grown in rpmi 1640 supplemented with 10 % fetal bovine serum is used as the target cell line in the assay . cells ( 100 μl ) are plated in microtest plate wells at a concentration of 10 5 cells per ml in the presence of varying concentrations of inhibitor . the cells are incubated at 37 ° c . in a humidified co 2 incubator . five days later , 20 μl of mtt ( 5 mg / ml in rpmi 1640 , sonicated , 0 . 2 micron filtered , and stored at 4 ° c .) is added to each well . after 4 hours additional incubation at 37 ° c ., 60 μl of triton - x is added to each well and thoroughly mixed to aid the solubilization of the crystals . absolute ethanol ( 5 μl ) is added to each well to remove bubbles and the resulting mixture is incubated for 30 minutes at 60 ° c . and immediately read on a plate reader ( dynatech ) at a wavelength of 570 nm . data from this assay are used to generate a nonlinear regression analysis which yields an ec 50 . table i______________________________________ rt inhibitioncompound other (% @ 10of example r . sup . 1 substituents * μg / ml ) ______________________________________1 ch . sub . 2 ch . sub . 2 ch . sub . 3 1002 ch ( ch . sub . 3 ). sub . 2 1003 ch . sub . 2 ch . sub . 2 ch . sub . 3 z ═ s 944 h x ═ s 435 ch . sub . 2 ch . sub . 2 ch . sub . 3 x ═ s 986 ch . sub . 3 2 - nh . sub . 2 , 7 - ch . sub . 3 1007 h 228 ch . sub . 2 ch . sub . 3 1009 ch . sub . 2 ch ═ ch . sub . 2 9810 ch . sub . 3 9311 ch . sub . 2 c ( ch . sub . 3 )═ ch . sub . 2 7712 coch . sub . 2 ch . sub . 3 4413 ch . sub . 2 conh . sub . 2 4 - och . sub . 3 3214 h 2 - nh . sub . 2 8515 ch . sub . 2 conh . sub . 2 2 - nh . sub . 2 8216 ch . sub . 2 ch . sub . 3 2 - nh . sub . 2 10017 h 2 - nh . sub . 2 , 8 - cl 6718 h 2 - nh . sub . 2 , 8 - ch . sub . 3 7119 ch . sub . 2 ch . sub . 3 2 - nh . sub . 2 , 8 - ch . sub . 3 10020 ch . sub . 2 ch . sub . 3 2 - nh . sub . 2 , 8 - cl 9521 h 7 - nh . sub . 2 322 ch . sub . 3 2 - nh . sub . 2 9923 h 3 - nh . sub . 2 2524 ch . sub . 3 7 - nh . sub . 2 4425 ch . sub . 3 3 - nh . sub . 2 2626 ch . sub . 3 2 - nhch . sub . 3 7527 ch . sub . 2 ch ═ ch . sub . 2 2 - nh . sub . 2 10028 h 2 - nhch . sub . 3 3029 ch . sub . 2 ch ═ ch . sub . 3 7 - nh . sub . 2 8930 h 2 - nh . sub . 2 , 7 - ch . sub . 3 9631 ch . sub . 2 conh . sub . 2 7 - ch . sub . 3 7732 ch . sub . 3 7 - och . sub . 2 ch . sub . 3 7733 ch . sub . 2 ch . sub . 3 7 - och . sub . 2 ch . sub . 3 7434 ch . sub . 2 sch . sub . 3 10035 ch . sub . 2 soch . sub . 3 5836 ch . sub . 2 ch . sub . 2 sch . sub . 3 8537 ch . sub . 2 sch . sub . 3 3 - cl 9838 ch . sub . 2 ch . sub . 2 sch . sub . 3 3 - cl 8339 ch . sub . 2 ch . sub . 2 sch . sub . 3 7 - ch . sub . 3 9540 ch . sub . 2 soch . sub . 3 7 - ch . sub . 3 7741 ch . sub . 2 ch . sub . 2 sch . sub . 3 2 - cl 4942 h 7 - och . sub . 2 ch . sub . 3 3043 ch . sub . 2 co . sub . 2 ch . sub . 2 ch . sub . 3 7644 h 7 - och . sub . 3 2745 h 1 , 7 -( och . sub . 3 ). sub . 2 4446 ch . sub . 2 ch . sub . 2 f 9347 cf ═ chi 9248 ch . sub . 2 cf . sub . 3 8749 ch . sub . 3 x ═ s 9450 ch . sub . 2 conh . sub . 2 x ═ s 66______________________________________ * x and z are each oxygen unless otherwise noted . table ii______________________________________compound t - cell culture cytotoxity assayof example assay (% inhibition ) ( ec . sub . 50 ) ______________________________________1 93 205 nt 10______________________________________ note : nt = not tested the following examples further illustrate the present invention and will enable others skilled in the art to understand the invention more completely . it should be understood , however , that the invention is not limited to the particulars given in the examples . to a solution of 3 . 0 grams of 10h - dibenz [ b , f ][ 1 , 4 ] oxazepin - 11 - one in 50 ml of dry dimethylformamide was added 0 . 82 grams of a 50 % dispersion of sodium hydride in mineral oil . the resulting mixture was stirred for one hour and 3 . 7 grams of 1 - bromopropane was then added slowly . the reaction mixture was stirred for 3 hours and the excess sodium hydride decomposed by the addition of ice . after further dilution with water the product was extracted with ether , dried ( anhydrous sodium sulfate ) and concentrated . the resulting oil was purified on a silica gel column ( ethyl acetate / hexane , 1 : 4 ) to provide 3 . 1 grams ( 87 % of theory ) of 10 - propyl - 10h - dibenz [ b , f ][ 1 . 4 ] oxazepin - 11 - one as a colorless oil . following the proceedure described in example 1 , 3 . 0 grams of 10h - dibenz [ b , f ][ 1 , 4 ] oxazepin - 11 - one and 3 . 7 grams of 2 - propyl bromide were reacted in the presence of 0 . 82 grams of a 50 % dispersion of sodium hydride in mineral oil and 50 ml of dry dimethylformamide . the product was purified on the silica gel column ( ethyl acetate / hexane , 1 : 4 ). the resulting oil was dissolved in petroleum ether which on standing gave 1 . 33 grams ( 30 % of theory ) of 10 - isopropyl - 10h - dibenz [ b , f ][ 1 , 4 ] oxazepin - 11 - one as colorless needles , m . p . 102 °- 103 ° c . a mixture of 2 . 45 grams of 10 - propyl - 10h - dibenz [ b , f ][ 1 , 4 ] oxazepin - 11 - one , prepared as in example 1 , 2 . 02 grams of lawesson ( s ) reagent and 50 ml of toluene was refluxed for 5 hours . the solvent was then removed in vacuo to give a yellow oil which was purified on a silica gel column ( ethyl acetate / hexane , 1 : 5 ) to give 1 . 05 grams ( 57 % of theory ) of 10 - propyl - 10h - dibenz [ b , f ][ 1 , 4 ] oxazepin - 11 - thione as a yellow oil . a mixture of 1 . 4 grams of 2 - aminothiophenol , 2 . 5 grams of 2 - iodobenzoic acid , 0 . 5 grams of copper bronze , 2 . 6 grams of potassium hydroxide and 15 ml of water were refluxed under nitrogen for 51 / 2 hours . the mixture was then filtered and the filtrate acidified with concentrated hydrochloric acid , stirred for 1 hour and the solid collected by suction filtration . after washing with ethanol and diethyl ether , the solid was dried in vacuo to give 2 . 1 grams ( 75 % of theory ) of 2 -[( 2 &# 39 ;- aminophenyl ) thiol ] benzoic acid as an off white powder , m . p . 220 ° c . ( dec .) suitable for use in the next reaction . the amino acid described above was heated neat to 200 °- 230 ° c . for 4 hours . crystallization from ethyl acetate provided 0 . 22 grams ( 23 % of theory ) of dibenz [ b , f ][ 1 , 4 ] thiazepin - 11 ( 10h )- one as a light beige crystalline power , m . p . 258 °- 259 ° c . 0 . 14 grams of a 50 % dispersion of sodium hydride in mineral oil was added to a solution of dibenz [ b , f ][ 1 , 4 ] thiazepin - 11 ( 10h )- one in 10 ml of dimethylformamide . after 15 minutes the mixture was warmed to 50 ° c . and stirred under argon for one hour . after allowing the mixture to cool to room temperature , 0 . 34 grams of 1 - bromopropane was added slowly . the reaction mixture was stirred for 5 hours , after which time it was quenched with water and the product extracted with diethyl ether . the extracts were washed with a saline solution , dried ( anhydrous magnesium sulfate ) and concentrated to give a colorless oil which was purified by column chromatography ( ethyl acetate / hexane , 5 %, 10 %, 20 %) to provide 0 . 37 grams ( 91 % of theory ) of 10 - n - propyl - dibenz [ b , f ][ 1 , 4 ] thiazepin - 11 ( 10h )- one as a white crystalline solid , m . p . 103 °- 104 . 5 ° c . a mixture of 20 . 2 g ( 0 . 1 mole ) of 2 - chloro - 5 - nitrobenzoic acid and 8 . 4 ml thionyl chloride in 100 ml of dioxane was refluxed for 90 minutes . the solution was cooled to 50 ° c . and added to a suspension of 12 . 3 g ( 0 . 1 mole ) of 2 - amino - 5 - methyl phenol in 50 ml of water . after about half of the acid chloride was added , 8 . 2 g sodium acetate was added , then the rest of the acid chloride was added dropwise . the reaction mixture was stirred for 45 minutes at 50 ° c . ice water was added to the mixture with stirring , and the resulting precipitate was filtered and washed with water . the filter cake was dissolved in 150 ml of water containing 4 g of naoh and stirred for 1 hour at 85 °- 95 ° c ., then cooled to room temperature . the cooled solution was acidified with hcl , filtered , and washed with water . the precipitate was dried to yield 11 . 3 g ( 42 %) of 2 - nitro - 7 - methyldibenz [ b , f ][ 1 , 4 ] oxazepin - 11 ( 10h )- one , mp 264 °- 266 ° c . to a warm solution of 4 . 05 g potassium methoxide in 80 ml of t - butanol and 80 ml of dioxane was added 13 . 5 g ( 0 . 05 mol ) of 2 - nitro - 7 - methyldibenz [ b , f ][ 1 , 4 ] oxazepin - 11 ( 10h )- one . the resulting suspension was warmed to 60 ° c . for 15 minutes . initially everthing went in solution , after several minutes a crystalline precipitate formed . to the stirred suspension was added 3 . 75 ml of iodomethane . the reaction mixture was stirred for 4 hours at 60 ° c ., then concentrated to dryness in vacuo . the dark residue was suspended in water , acidified with acetic acid , concentrated in vacuo and then recrystallized from propanol to give 8 . 5 g ( 60 % yield ) of 2 - nitro - 7 , 10 - dimethyldibenz [ b , f ][ 1 , 4 ]- oxazepin - 11 ( 10h )- one mp 156 °- 159 °. a mixture of 8 . 5 g ( 0 . 03 mole ) of 2 - nitro - 7 , 10 - dimethylbenzoxazepin - 11 ( 10h ) one in 100 ml of ethanol and 3 g raney nickel was hydrogenated at 50 ° c . and 50 atm . the hydrogenation mixture was heated to boiling and filtered . the filtrate was cooled and the crystalline precipitant filtered off to give 4 . 4 g ( 58 %) of 2 - nitro - 7 , 10 - dimethyldibenz [ b , f ][ 1 , 4 ] oxazepin - 11 ( 10h ) - one , mp 183 °- 185 ° c . using synthetic methods analogous to those described above , the following compounds were prepared : ______________________________________ meltingcompound other pointsof example r . sup . 1 substituents * (° c . ) ______________________________________7 h 211 - 2138 ch . sub . 2 ch . sub . 3 53 - 549 ch . sub . 2 ch ═ ch . sub . 2 89 - 9110 ch . sub . 3 81 - 8211 ch . sub . 2 c ( ch . sub . 3 )═ ch . sub . 2 83 - 8412 coch . sub . 2 ch . sub . 3 78 - 8013 ch . sub . 2 conh . sub . 2 4 - och . sub . 3 238 - 24014 h 2 - nh . sub . 2 200 - 20215 ch . sub . 2 conh . sub . 2 2 - nh . sub . 2 224 - 22516 ch . sub . 2 ch . sub . 3 2 - nh . sub . 2 165 - 16617 h 2 - nh . sub . 2 , 8 - cl 266 - 26718 h 2 - nh . sub . 2 , 8 - ch . sub . 3 169 - 17019 ch . sub . 2 ch . sub . 3 2 - nh . sub . 2 , 8 - ch . sub . 3 114 - 11520 ch . sub . 2 ch . sub . 3 2 - nh . sub . 2 , 8 - cl 255 ( dec ) ( hcl salt ) 21 h 7 - nh . sub . 2 na22 ch . sub . 3 2 - nh . sub . 2 133 - 13623 h 3 - nh . sub . 2 287 - 28924 ch . sub . 3 7 - nh . sub . 2 184 - 18625 ch . sub . 3 3 - nh . sub . 2 187 - 18926 ch . sub . 3 2 - nhch . sub . 3 132 - 13427 ch . sub . 2 ch ═ ch . sub . 2 2 - nh . sub . 2 156 - 15828 h 2 - nhch . sub . 3 175 - 17729 ch . sub . 2 ch ═ ch . sub . 3 7 - nh . sub . 2 112 . 5 - 115 . 530 h 2 - nh . sub . 2 , 7 - ch . sub . 3 184 - 186 . 531 ch . sub . 2 conh . sub . 2 7 - ch . sub . 3 225 - 22632 ch . sub . 3 7 - och . sub . 2 ch . sub . 3 oil33 ch . sub . 2 ch . sub . 3 7 - och . sub . 2 ch . sub . 3 oil34 ch . sub . 2 sch . sub . 3 88 - 8935 ch . sub . 2 soch . sub . 3 158 - 16036 ch . sub . 2 ch . sub . 2 sch . sub . 3 78 - 8037 ch . sub . 2 sch . sub . 3 3 - cl 85 - 8738 ch . sub . 2 ch . sub . 2 sch . sub . 3 3 - cl 87 - 8839 ch . sub . 2 ch . sub . 2 sch . sub . 3 7 - ch . sub . 3 54 - 5540 ch . sub . 2 soch . sub . 3 7 - ch . sub . 3 99 - 10341 ch . sub . 2 ch . sub . 2 sch . sub . 3 2 - cl 101 - 10242 h 7 - och . sub . 2 ch . sub . 3 180 - 18243 ch . sub . 2 co . sub . 2 ch . sub . 2 ch . sub . 3 82 - 8444 h 7 - och . sub . 3 202 - 20345 h 1 , 7 -( och . sub . 3 ). sub . 2 249 - 25246 ch . sub . 2 ch . sub . 2 f 71 - 72 . 547 cf ═ chi 133 - 13648 ch . sub . 2 cf . sub . 3 109 - 11049 ch . sub . 3 x ═ s na50 ch . sub . 2 conh . sub . 2 x ═ s na______________________________________ * x and z are each oxygen unless otherwise noted . ______________________________________example acapsules or tabletsingredients quantity______________________________________a - 1compound of example 1 50 mgstarch 160 mgmicrocrys , cellulose 90 mgsodium starch gluctate 10 mgmagnesium stearate 2 mgfumed colloidal silica 1 mga - 2example 1 50 mgdicalcium phosphate 160 mgmicrocrys . cellulose 90 mgstearic acid 5 mgsodium starch glycolate 10 mgfumed colloidal silica 1 mg______________________________________ the compound of example 1 is blended into a powder mixture with the premixed exipient materials as identified above with the exception of the lubricant . the lubricant is then blended in and the resulting blend compressed into tablets or filled into hard gelatin capsules . ______________________________________example bparenteral solutionsingredients quantity______________________________________compound of example 1 500 mgethanol 25 mlwater for injection q . s . to 100 ml______________________________________ compound of example 1 is added to the ethanol and mixed until the solution is clear . water is added and the resulting solution is then filtered into the appropriate vials or ampoules and sterilized by autoclaving . ______________________________________example cnasal solutionsingredients quantity______________________________________compound of example 1 500 mgpropylene glycol 30 mlbenzalkonium chloride 200 mgedta 200 mgwater q . s . to 100 ml______________________________________ the excipient materials are mixed and thereafter the compound of example 1 is added and mixing is continued until the solution is clear . the water is added and the resulting solution is then filtered into the appropriate vials or ampoules .

disclosed is the use of dibenzoxazepin - 11 - ones and - thiones in the prevention and treatment of aids , arc and related disorders associated with hiv infection .

with reference now to fig1 lure rod 10 comprises a first rod section 12 located at the tip and a second rod section 14 located on the bottom end of the rod with the first and the second rod sections connected in parallel . the second rod section 14 has a line introduction hole 16 for inserting a line from a reel 18 into a wide section formed as a line introduction section 20 in the front end section and also has a reel seat 22 and a grip 24 behind the line introduction hole . the second rod section 12 comprises an internal path 26 leading from the line introduction hole 16 to a tip opening 28 . an internal body 30 forming a line insertion and passage path 32 can be inserted from a rear end opening 34 in the first rod section 12 . the line introduction section 20 can be connected to or disconnected from the rear end of the first rod section 12 in parallel , and is formed to have a large width . the line introduction hole 16 that is longitudinally long is formed in the longitudinal center of the line introduction section 20 , an internal space is formed inside the line introduction section 20 , and a path leading to the internal path 26 is formed in a position in front of the line introduction hole 16 . the rear end of this path enters the internal space from the front end of the line introduction hole 16 and can be seen externally from the line introduction hole 16 . the internal space formed between the rear end of the space 26 and the line introduction hole 16 is used as a communication space 36 for guiding to the rear end of the space 26 ( the internal space for inserting and passing a line ) a line introduced from the line introduction hole 16 . an internal - external through hole 38 is formed on the lower surface of the line introduction section 20 and located symmetrically 180 ° from to the line introduction hole 16 . it is shaped like a longitudinally long hole and has shorter longitudinal length and width than the line introduction hole 16 . dirt entering the line insertion and passage path 32 adheres to a line and is then moved to the communication space 36 , where it falls and is then discharged through the internal - external through hole 38 . as a result , dirt adhering to a line rolled up by the reel 18 can be reduced . fig1 , and 4 show that the internal body 30 is installed in the rear half section of the first rod section 12 and has the following configuration . a plurality of units each of which comprises a cylindrical section 40 where the line support 42 is installed , and a connecting rod section 44 extending rearwardly from the cylindrical section 40 like a cantilever , are bonded and connected together so as to have a length equivalent to a single fishing rod . the cylindrical section 40 has formed in its center a through hole 46 into which the line support is inserted and loaded , with the connecting rod section 44 extending from a position above the shaft center of the through hole 46 . this positioning of the connecting rod section 44 allows water entering the rod to flow unaffected by the connecting rod section 44 . in addition to this configuration , the formation of the longitudinal through hole 48 in the cylindrical section 40 allows water entering the rod to be guided to the internal - external through hole 38 for draining , where the water is discharged . the connecting rod section 44 is formed by removing most of the corresponding wall of the cylindrical section 40 and assumes an approximate crescent cross section . the internal body 30 generally comprises thermoplastic resin without reinforced fibers but may also comprise rubber . in this embodiment , it is formed of a material more flexible than the rod 10 . the thermoplastic resins useful in the present invention include polyethylene , polypropylene , nylon , abs , and most preferably is a material having high water repellency . on the other hand , the line support 42 housed in the cylindrical section 40 is formed like a ring by forming on the front surface of a block body a small diameter hole 50 for inserting and passing a line and constructing a tapered line guide surface 52 that gradually widens rearwardly from the opening of the small diameter hole 50 , as shown in fig2 . the material for the line support 42 includes hard materials such as ceramics and cermets ( sintering materials ) but may be a flexible material such as resin . the line support 42 is loaded into the through hole 46 of the cylindrical section 40 , and the rear end of the connecting rod section 44 located in front of the cylindrical section 40 is inserted into the through hole 46 to contact the line support 42 in such a way that the line support 42 can be positioned appropriately . the connecting rod section 44 and the cylindrical section 40 are then bonded and fixed together , and the internal body is incorporated into the line support 42 while assembled . once a plurality of units are connected to form the internal body 30 , a line insertion and passage path 32 is formed from the small diameter hole for line insertion and passage 50 through the line guide surface 52 and the shaft center position of the connecting rod section 44 . the connecting and fixation structure for each unit may comprise various fixation structures using fitting or bolting . fig2 shows that the first rod section 12 has a top guide installed at its rear end and that the top end 54 has installed at its tip a ring for guiding a line , with drain holes 56 formed behind the ring 58 . the tip structure of the first rod section 12 is described . fig2 shows that the line support 42 and the retainers 60 for retaining the line support 42 are installed in front of the internal body 30 . the line support 42 is a long spiral 62 along the shaft center of the rod , and the lower inner surface of the spiral 62 is used as a line support section 50 for substantially supporting a line . this allows the line support sections 50 to be arranged at the pitch of the spiral 62 . on the other hand , a plurality of retainers 60 are constructed along the shaft center of the spiral 62 to retain the line support 42 . the retainer 60 is like a cylinder and retains the line support within its through hole 64 located in its center . these retainers 60 are arranged at a predetermined pitch 66 that is larger than the line support pitch 68 of the line support 50 . this allows the line support pitch 68 to be small and prevents a line from falling from between adjacent line support sections 50 and contacting the inner surface of the rod or water . another tip structure for the first rod section 12 is shown in fig4 . fig4 shows that a line support 42 and a retainer 60 for retaining the line support 42 are installed in front of the above internal body 30 . the line support 42 comprises a long spiral 62 along the shaft center of the rod and a thin cylinder 70 externally fit to the spiral 62 for tightening and retaining the spiral 62 . the spiral 62 has a small element wire diameter and a relatively high flexibility , and the thin cylinder 70 comprises a thermally contracting tube and is externally fit to the spiral 62 while adhering thereto . this enables the spiral 62 to constantly maintain its shape despite its small diameter provided to improve its flexibility in order to address the small diameter of the tip rod . the spiral 62 formed in this manner is a cylinder of a specified outer diameter along its length . an opener 72 is attached to the rear end of the thin cylinder 70 , and a cylindrical retainer 60 externally fit to the opener 72 for retaining the line support 42 is constructed inside the first rod section 12 . the front end of the line support 6 directly contacts the tapered inner surface of the first rod section 12 and is retained therein . this construction serves to maintain a space between the section behind the front end of the line support 42 and the inner circumferential surface of the first rod section 12 . the retainer 60 of the configuration according to this invention can be configured as follows . it may be like a simple cylinder as shown in fig5 . as shown in fig6 a cylinder - like body may be used as a base , and two engagement sections 74 , may protrude from the outer circumferential surface of the body . an engagement groove for engaging the engagement sections 74 with the inner surface of the first rod section 12 is constructed in the shaft center direction of the first rod section 12 so that the retainer 60 can be moved in the shaft center direction of the first rod section 12 and that the outer ends of the engagement sections 74 can be fit in a predetermined position to the bottom surface of the engagement groove to fix the retainer 60 . however , this is not shown in the drawings . as shown in fig7 one of the engagement sections 74 may be formed in a depressed section 76 . the depressed section 76 thus forms a path for moving water . as shown in fig8 an insertion and passage hole 76 for inserting and passing the line support 42 may be formed in the shaft center of the block body , and a guidance inclined surface 78 may be constructed in one of the openings of the insertion and passage hole 80 . the line support 60 of the configuration according to this invention may be configured as follows . the line support section 82 may be configured as follows . as shown in fig9 a plurality of ring - like line support sections 82 may be arranged and connected by the connecting rod section 84 as a single unit . the reference numeral 62 in the figure designates a line support 42 constituted by configuring the line support sections 82 as a single unit using a connecting rod section 84 and covering the unit with a thin cylinder 70 such as a thermally contracting tube . as shown in fig1 , the ring - like support sections 82 and the connecting rod section 84 may be formed of a single raw material , thus forming a unitary structure . as shown in fig1 , the line support sections 82 of the above configuration may be formed of rings with a plurality of turns . as shown in fig1 , the ring - like line support sections 82 need not form a closed loop but may form an open loop that opens in the connecting position of the connecting rod section 84 . the connecting rod section 84 may be configured as follows . it may be circumferentially located in three positions , as shown in fig1 it may be laterally located in two positions , as shown in fig1 . as shown in fig1 , it may be formed like bellows having raised and depressed surfaces inside , with the raised surfaces used as line support sections 82 . as shown in fig1 , it may be a cylindrical body with many small holes drilled on its circumferential surface . the cylindrical body may comprise punching metal . it may comprise resin fibers laced up as a mesh . however , this is not shown in the drawings . other embodiment structures are described as follows . as shown in fig1 , the retainer 60 for retaining the line support 42 may be located in a plurality of positions including those in the large diameter section and the small diameter section . in this case , such an arrangement can be provided instead of the line support shown as the spiral 62 integrated using the thermally contracting tube 70 . for example , the line support 42 may comprise the above single spiral . as shown in fig1 , a plurality of small diameter rings and large diameter rings may be alternatively located adjacent to each other on the same shaft center , and then connected together using the connecting rod section . in this case , the small diameter rings form a line support 82 , and the large diameter rings form a retainer 60 contacting the inner circumferential surface of the first rod 12 . as shown in fig1 , to support the spiral line support 42 spaced from the inner surface of the rod , the line support may be supported by the retainer 60 at the rear end of the rod , while , at the front end of the rod , it may be received by a swelled section 86 formed by inwardly swelling part of the inner surface of the rod . all the above embodiments can be used for rods other than the tip rod such as an intermediate rod .

a pass - through fishing rod comprising a line support and retainers prevents a fishing line from contacting the inner surface of a rod or water adhering to the inner surface and enables the line to be drawn out and rolled up smoothly . the line support pitch of the line support supporting a fishing line is smaller than the retention pitch of the retainers for retaining the line support spaced from the inner surface of the rod .

in one embodiment of the inventive system , the glucose concentration is measured in the eye , by a photoacoustic assay such as that described in u . s . pat . no . 6 , 846 , 288 ( e . g . at col . 13 , line 62 to col . 18 , line 49 ) or in u . s . pat . no . 6 , 403 , 904 , each of which is incorporated by reference in their entirety . as shown in fig1 , the probe module 60 includes an objective lens structure 35 , which is coupled to a light source 45 via a fiber optic connection or other light transmitter . light source 45 provides light at a wavelength which is preferentially absorbed by glucose . alternatively , the light source may be incorporated into the probe module 60 . the light source 45 may be a laser , laser diode or superluminescent diode ( sld ), as appropriate for generating the desired light wavelength and intensity . the light may be delivered as pulses or as modulated radiation , the probe module 60 further contains an ultrasound transducer 50 to detect the photoacoustic waves that are generated as a result of the absorption of energy from the light emitted by the objective lens structure 35 . the ultrasound transducer 50 is in contact with the eye 30 or an eyelid drawn over the eye . as light is delivered as pulses or as modulated radiation ( as elaborated in the above - referenced u . s . pat . nos . 6 , 846 , 288 and 6 , 403 , 904 ), pulses or modulating acoustic signals are generated and returned to the ultrasound transducer 50 in probe module 60 . as noted , it is expected that substantially superior results , in repeatability and ease of calibration , will be achieved in the eye than are achieved in soft tissue as proposed by the &# 39 ; 288 and &# 39 ; 904 patents . it will be appreciated that localization of the source of photoacoustic signals may be achieved in various manners . first , localization may be accomplished by directing the beam from objective lens structure 35 in specific directions , by moving that structure with micromechanical actuators as shown diagrammatically at 27 in fig1 , thus targeting a particular line of points in the eye . furthermore , by suitable optics included in objective lens structure 35 , the focal point of the emitted light may be moved within the eye to a desired point , such as a point along the retina vasculature , to selectively generate acoustic signals at that desired point . because the eye is optically transmissive relative to soft tissue , beam focusing and beam directing are likely to be more accurately performed in the eye , than is the case is soft tissue elsewhere in the body . to further assist in directionally capturing the photoacoustic signals generated within the eye , a directional transducer array may be used as transducer 50 , to control the directionality of reception of ultrasonic energy , thus further localizing upon a desired source of thermoacoustic signals . thus , by targeting the focal point of the illuminating light , and also directionally targeting the reception of ultrasonic signals by the transducer array , thermoacoustic signals from a particular location , such as along the retina , may be specifically targeted . mapping of patient eye structures is useful for analysis of macular edema , macular holes , glaucoma , various retinal diseases , neuroophthalmology , the anterior segment , and normal eye conditions . for such applications , it will be appreciated that the ultrasound transducer 50 may transmit and receive ultrasound waves ; when transducer 50 transmits waves , reflected ultrasound waves may be used for imaging of eye structures , as is a known use of ultrasound for imaging . eye structures localized with ultrasound may then be targeted by the optical system and transducer 50 for photoacoustic analysis . the combined use of sensors for ultrasound imaging and thermoacoustic reception is explained further in u . s . pat . no . 6 , 490 , 470 , which is hereby incorporated herein by reference in its entirety . it will also be appreciated that the apparatus shown in fig1 may be adapted to analyze glucose with reflected or transmitted ultrasound , i . e ., ultrasound that passes through an irradiated area in the eye may be captured and its properties analyzed to determine glucose concentration , as discussed in u . s . pat . no . 6 , 846 , 288 . for such an application , an ultrasonically reflective structure may be utilized to reflect ultrasound transmitted into the eye so that the reflected ultrasound may be analyzed . the skull may be utilized for this purpose , or another reflector may be temporarily positioned within the eye socket for this purpose . alternatively , an ultrasound emitter may be placed adjacent to the eye within the eye socket to generate ultrasound that may be received by a directionally - oriented receiver after passing through an area subject to illumination by light source 45 . in alternative embodiments of the invention , glucose concentrations within the eye may be measured in conjunction with a reflection interferometry method , such as a short coherence reflection interferometry method as generally described ( for soft tissue ) in u . s . pat . no . 5 , 710 , 630 ( col . 10 , line 20 to col . 14 , line 27 ). in this system , as shown in fig2 , the probe contains a superluminescent diode ( sld ) light source 45 that transmits a wavelength of , e . g ., 1300 nm , delivered to the imaging site through optical fiber and a coupler 70 , into the eye 30 . light is also coupled from coupler 70 to a reference path 75 , from which it is reflected to create interferometry with the reflections from the eye 30 at a photodetector 80 . movement of the objective lens 35 within the probe 60 as shown at 27 permits illumination of specific eye features . the characteristics thereof may then be detected from the changing interferometry between the reflected light received from lens 35 and reference path 75 , as described in u . s . pat . no . 5 , 710 , 630 . by using a low - coherence - length light source 10 and measuring the interference between light backscattered from a tissue and from a mirror in the reference path 75 , the distance and magnitude of optical scattering within the tissue is measured . in a combined photoacoustic and interferometric system as illustrated in fig3 , the optical interferometry section generates light waves that reflect off the internal microstructure and also cause localized photoacoustic signal generation . interferomatic techniques extract the reflected optical signals from the infrared light and the output , measured by an interferometer , is processed to produce glucose measurements as well as potentially cross sectional or 3 - dimensional images of the target site . in this combined embodiment , probe module 60 also includes an ultrasound transducer 50 and glucose detector system 85 of the kind discussed with reference to fig1 . a processor 90 attached to interferometry detector 80 and to glucose detector 85 combines the resulting signals from each . scanning the light beam across the tissue produces a cross - sectional image by the signal processor 90 , while processor 90 records the axial reflectance profiles at each transverse position . processor 90 also records glucose measurements at each location generated by inteferometric methods and from photoacoustic data generated in response to light illumination . the result is a multi - dimensional representation of the optical backscattering of the tissue &# 39 ; s cross - section , which displays as a gray - scale or false - color image , and a superimposable measure of glucose . while various embodiments have been chosen to illustrate the invention , it will be understood by those skilled in the art that various modifications can be made without departing from the spirit and scope of the invention as defined in the following claims .

in one aspect , the invention features a method and device for measuring blood concentration of a substance such as glucose in the aqueous humor by illuminating the aqueous humor with a light source at a frequency that is absorbed by the substance to the measured , and then sensing photoacoustically generated sound waves originated within the aqueous humor as a consequence of illumination by the light source . the blood concentration can be estimated from the amplitude of the sound waves received . the method may be combined with other optical techniques for glucose measurement and / or with optical or ultrasonic techniques for topographic mapping of eye structures .

fig1 illustrates a collapsible table 10 having a support frame 12 , a pair of expansion leaves 14 pivotally connected to opposing sides of the support frame 12 , and a locking mechanism 16 for selectively maintaining the expansion leaves 14 in an expanded configuration 18 . in fig1 , the collapsible table 10 is shown in an expanded configuration 18 wherein the support structure 12 is expanded horizontally by a configurable truss 20 . the expansion leaves 14 extend horizontally from opposing sides 22 , 24 of the support structure 12 . the locking mechanism 16 extends from the support frame 12 to the expansion leaf 14 to maintain the expansion leaf 14 in the expanded configuration 18 . each leaf 14 is supported by a locking mechanism 16 for maintaining a support arm 26 in axial alignment with a corresponding extension arm 28 , to support the leaf 14 . in fig1 , the collapsible table 10 is shown in a collapsed configuration 30 wherein the support structure 12 is collapsed horizontally about the configurable truss 20 . the expansion leaves 14 are pivoted to extend vertically from opposing sides 22 , 24 of the support structure 12 . the locking mechanism 16 is folded at a pivotable junction 32 between the support arm 28 and extension arm 28 such that the support arm 26 and extension arm 28 are substantially vertical . a stop bar 34 extends from the support structure 12 for maintaining the support arm 26 and extension arm 28 in the vertical position . the locking mechanism 16 is shown in detail in fig2 - 8 . each locking mechanism 16 has a support arm 26 pivotally connected to the support structure 12 , and an extension arm 28 pivotally connected at opposing ends 36 , 38 to the support arm 26 and the expansion leaf 14 , respectively . the pivotable junction 32 is formed where the support arm 26 is pivotally connected to the extension arm 28 . the extension arm 28 has a slot 40 configured to slidably receive a pin 42 that extends from the support arm 26 . the pin 42 cooperates with the support arm 26 to maintain the extension arm 28 in axial alignment with the support arm 26 , when in the expanded configuration 18 . the locking mechanism 16 allows the expansion leaves 14 to be expanded to a horizontal position 44 or collapsed to a vertical position 46 . the support arm 26 is pivotally connected at a first end 48 to a lower portion 52 of the support frame 12 , on a lower cross bar 54 on the side 22 or 24 of the support frame 12 . the support arm 26 extends to a second end 50 that is pivotally connected to a first end 36 of the extension arm 28 . the extension arm 28 is pivotally connected at a second end 38 to the underside 56 of the expansion leaf 14 . a locking joint 32 is formed where the extension arm 28 is pivotally connected to the support arm 26 . the first end 36 of the extension arm 28 has a slot 40 for receiving a pin 42 that extends from the second end 50 of the support arm 42 . in the extended configuration 18 , the pin 42 and support arm 26 cooperate to maintain the extension arm 28 in axial alignment with the support arm 26 . the slot 40 has an outer , locking end 58 , and an inner , pivoting end 60 . when the pin 42 engages the outer end 58 of the slot 40 , the extension arm 28 slides into a channel 62 in the support arm 26 such that the bottom 64 of the channel 62 engages the extension arm 28 . when the pin 42 slides within the slot 40 to the inner , pivoting edge 60 , such as by sliding the extension arm 28 away from the support arm 26 along the axis of alignment , the extension arm 28 is able to pivot about the pin 42 with respect to the support arm 26 , toward a collapsed configuration 30 . the channel 62 of the support arm 26 is configured such that , in the extended configuration , the channel 62 faces away from the support structure 12 . the first end 36 of the extension arm 28 is received within the channel 62 . an aperture 66 is defined by a portion of the bottom 64 of the channel 62 . a brace 68 is positioned at the second end 50 of the support arm 26 so as to prevent the pivoting of the pivot point 32 in a direction away from the support structure 12 beyond the axial alignment of the support arm 26 and extension arm 28 . the aperture 66 allows the extension arm 28 to pivot within the channel 62 when moved to a collapsed configuration 30 . the extension arm 28 is slid away from the support arm 26 such that the pin 42 engages the pivot end 60 of the slot 40 . a force f may then be applied to the pivot area 32 to bend the pivot area 32 and move the pivot area 32 toward the support structure 12 . as the pivot area 32 moves toward the support structure 12 , the second end 50 of the support arm 26 moves toward the stop bar 34 , with the first end 36 of the extension arm 28 . the second end 38 of the extension arm 26 moves downwardly with the outer end 70 of the expansion leaf 14 . the support structure 12 includes four upright members 72 , 74 , a front pair 72 and a rear pair 74 . as shown in fig9 , a configurable truss 20 , 21 extends between the front pair 72 , and identically between the rear pair 74 of upright members . the configurable truss 20 , 21 has two crossing arms 76 . each crossing arm extends angularly from a first upright member 72 , 74 to a second upright member 72 , 74 . the two crossing arms 76 are pivotally connected to each other by a pin 78 , forming a pivot point , between the pair of upright members 72 , 74 . the pin 78 extends from the front configurable truss 20 to the rear configurable truss 21 so as to maintain the trusses 20 , 21 in a mirrored relationship . each crossing arm 76 has one end 80 that is pivotally fixed to a portion of an upright member 72 , 74 , and another end 82 that is slidably extendible along the vertical length of the upright member 72 , 74 . the sliding end 82 is slidably connected to the upright member 72 , 74 by a guide member 84 that extends into a channel 86 within the upright members 72 , 74 . a stopper 88 is positioned within the channel 86 to limit the upward and downward distance that the crossing arm 76 can extend along the length of the upright member 72 , 74 . as shown , the lower end 90 of the crossing arms 76 are fixed to the lower portion 52 of the upright members 72 , 74 and the upper end 94 of the crossing arms 76 are able to slide along the vertical length of the upright members 72 , 74 . the configurable truss 20 , 21 is able to extend in the horizontal direction wherein the upper end 94 of the crossing arms 76 will slide downwardly along the vertical length of the upright member 72 , 74 . conversely , the configurable truss 20 , 21 is able to collapse in the vertical direction wherein the upper end 94 of each of the crossing arms 76 will slide upwardly along the vertical length of the upright member 72 , 74 . the support structure 12 has crossing bars 54 , 96 extending from the front pair of upright members 72 to the rear pair of upright members 74 such that the crossing bars 54 , 96 are parallel to each other and perpendicular to the configurable trusses 20 , 21 . as shown , there is an upper crossing bar 96 and a lower crossing bar 54 that extends from each of the front upright members 72 to a corresponding rear upright member 74 . a stop bar 34 extends vertically downwardly from each of the upper crossbars 96 . the stop bar 34 is able to engage the support arm 26 when in the collapsed configuration 30 to prevent pivoting of the support arm 26 in the direction of the stop bar 34 , wherein the support arm 26 abuts against the stop bar 34 . the expansion leaves 14 are pivotally connected to opposing ends 22 , 24 of the support structure 12 . a front portion 98 of the expansion leaf 14 is pivotally connected to a top portion 92 of the front upright member 72 and a rear portion 100 of the expansion leaf 14 is pivotally connected to a top portion 92 of the rear upright member 74 . the expansion leaves 14 have a generally planar top surface and are extendible from a collapsed configuration 30 wherein the expansion leaves 14 are substantially in a vertical position and an expanded configuration 18 wherein the expansion leaves 14 are in a substantially horizontal position . each of the upright members 72 , 74 of the support structure 12 may alternatively have wheels 102 , or instead may have terminating ends 104 . a removable center table piece 106 is configured to be received over the support structure 12 . the table piece 106 has two pairs of recesses 108 , 110 , each pair of recesses 108 , 110 for receiving one of the crossing bars 96 . the table piece 106 has a generally planar top surface 112 . the table piece 106 is maintained over the support structure 12 by the fitting of the recesses 108 , 110 over the corresponding crossing bars 96 . the table piece 106 maintains the configurable truss 20 , 21 in the expanded configuration 18 . in the expanded configuration 18 , the vertical height of the removable table piece 106 is lower than the vertical height of the expansion leaves 14 . it is contemplated that the table piece 106 is configured as a working table , such as for maintaining a miter saw thereon . alternatively , the table may be used for recreation . the table piece 106 may be removed by lifting it upwardly from position over the crossing bars 96 .

the present invention provides an expandable and collapsible table for use with a miter saw or for recreational use . a support structure includes a truss that is expandable and collapsible in the horizontal direction . a pair of expansion leaves are pivotally connected to opposing sides of the support structure . a support arm and extension arm work in combination with a locking mechanism to maintain the expansion leaves in the horizontal position or alternatively to allow the expansion leaves to collapse to a vertical position .

while referring to fig1 to 6 , a guiding device 1 which is removably fixed on an operating table 2 intended more particularly to surgical operations is described . said operating table 2 is of the type including a top 3 for supporting a patient , fixed on a wheeled trolley ( not shown ). advantageously , the top 3 of the operating table 2 is obtained by juxtaposing three parts 4 , 5 and 6 intended to respectively receive the top of the back , the bottom of the back and the hips , as well as the legs of the patient . the operating table 2 further includes side rails 7 , which are fixed on the side edges of the top 3 , and more particularly on the side edges of each part 4 to 6 through spacer 8 . the rails 7 thus positioned around the top 3 make it possible to hang accessories required by the surgeon during the surgical operation . the guiding device 1 according to the invention includes two guiding arms 9 and 10 extending in the same plane and symmetrical with respect to each other in a longitudinal direction . in order to simplify the reading of the description , the structure of the guiding arm 9 will only be described , the other guiding arm 10 being symmetrical to said guiding arm 9 . the guiding arm 9 has a free end provided with fastening means 11 enabling the removable fastening on either side of the operating table 2 on the rail 7 . the fastening means 11 are composed of one part 12 , having a low thickness , and being substantially triangular . they are dimensioned so as to enable the insertion thereof into the space defined by the rail and the operating table . more particularly , said part 12 includes an outer face 13 and an inner face , and three edges : an upper edge 14 composed of the upper edge of the part 12 , a front side edge 15 and a rear side edge 16 . the terms “ outer face ” and “ inner face ” mean the faces of the part 12 , when it is engaged into the space formed between the top 3 and the side rail 7 , respectively positioned towards the rail 7 or towards the top 3 . similarly , the terms “ front side edge ” or “ rear side edge ” mean the position of the edges with respect to the guiding arm 9 . advantageously , the junction between each of the edges is rounded , with a view to avoiding injuries to the user of the operating table as well as to the patient , more particularly during the handling of the guiding device during the mounting or the dismounting thereof . the front side edge 15 is positioned substantially perpendicularly to the guiding part 9 , the rear side edge 16 being slightly slanted and oriented opposite the front side edge 15 . according to the described embodiment , the front side edge 15 has a length greater than that of the rear side edge 16 , so that the upper edge 14 is also slightly sloping . the upper edge 14 is provided with a notch 17 oriented along an axis bb 1 substantially perpendicular to the plane of the part 12 . as will be described in the following , said notch 17 is so configured as to receive one of the spacers connecting a side rail 7 to the operating table 2 . said notch 17 includes a bottom 18 which is substantially circular and two side walls , a front side wall 19 and a rear side wall 20 having the shape of a hook . as will be seen in the following , the front side wall 19 acts as a spacer stop 25 . the outer face 13 of the part 12 further includes an extension which acts as a bearing lug 21 intended to block the guiding device 1 on the side rails 7 of the operating table 2 . in the described embodiment , the bearing lug has a substantially circular cross - section . it should be noted that this configuration is given as an example and that said bearing lug may have other shapes of cross - sections without leaving the scope of the present invention . according to the embodiment described , the bearing lug 21 is formed on the outer face 13 , close to the junction of the front side edge 15 and the upper edge 14 of the part 12 . advantageously , the bearing lug 21 extends perpendicularly to the part 12 , so as to have an axis substantially parallel to the axis of the spacer which will be received in the notch 17 ( or the axis bb 1 of the notch 17 ). the bearing lug 21 will further be so dimensioned as to be pressed on the upper surface of said rail 7 when the guiding arm 9 is inserted into the space formed between the top 3 and said rail , the notch 17 accommodating the spacer 8 of the relating rail . the principle for the fastening of the guiding device 1 according to the invention on the operating table 2 is illustrated in fig2 a , 2 b , 3 a , 3 b and 3 c . the guiding device 1 is shown in front of the top 3 , with the ends of the guiding arms 9 and 10 carrying the fastening means 11 sloping downwards . the bearing lugs 21 of the guiding means 9 and 10 are so positioned as to rest on the side rails 7 . the guiding arms 9 and 10 are then pushed into the space formed between the concerned part 4 of the top 3 and the side rails 7 which are associated therewith , the bearing lugs 21 sliding on the rails 7 until the front side wall 19 of each notch 17 comes into contact with the end spacer 80 connecting the rails 7 with part 4 ( motion illustrated in fig3 c by a double arrow ). according to a particularly advantageous configuration of the invention , the guiding arms 9 and 10 are made integral with each other , so as to enable a side flexion of each of said arms . thus , when acting on the flexion of the guiding arms 9 and 10 , the spacing between said arms is adapted to the inter - rails spacer which can vary from an operating table to another . the flexion of the arms further provides the advantage of enabling the fastening thereof , as well at the level of the part 4 of the top ( refer to fig1 ), or of the part 6 of the top which acts as the leg holders ( refer to fig6 ). and it is possible to move the table by driving it as well from the patient &# 39 ; s head side as from the feet side . this characteristic makes it possible to adapt the transport of the patient to specific needs . for example , it may be advantageous in some situations to move the operating table 2 via a guiding device 1 fixed at the level of the feet , so as to be able to leave the patient &# 39 ; s head close to the wall , in the recovery room , for example , and thus close to the medical assistance facilities . when the front side walls 19 of each notch 17 are in abutment against the spacers 80 , the guiding arm 9 and 10 are released . this action entails that said guiding arms 9 and 10 are lowered until they reach a substantially horizontal position . the blocking in this position is then obtained through the joint action of the bearing lugs 21 and the rear side wall 20 of the notches having the shape of a hook , since no stress is exerted then . as a matter of fact , the lowering of the guiding arms 9 and 10 leads to a pivoting motion of the later about the bearing lugs 21 which motion results in that the hooks are positioned about the corresponding spacers 80 . the height of the hooks is an essential feature for the hanging of the guiding arms 9 and 10 . as a matter of fact , the height determines the mounting and dismounting angle of said guiding arms 9 and 10 ( angle α ). the fastening of the guiding arms 9 and 10 on the side rails is thus obtained through a “ mingling ” about the rail spacers and the rails proper . as for the blocking , it is obtained by gravitation . the fastening will be further improved by the slightly curved shape of the notch 17 ( hook ) which composes the fastening means . then , the effort is exerted directly on the spacers which are integral with the table , when the guiding arms 9 and 10 are pulled or pushed . the removal of the guiding arm 9 and 10 will be obtained similarly with a reverse motion . in order to enable the adjustment of the position of the guiding arms 9 and 10 to the user &# 39 ; s size , and also to be able to use the guiding device 1 on an operating table 2 where the patient is in a sitting position , i . e . where one of the parts of the top 3 of the table has been lifted up , the possibility of adjusting the height of the position of the guiding arms 9 and 10 with respect to the operating table 2 , as illustrated in fig4 , 5 a and 5 b , is advantageously provided for . for this purpose , the part 12 is mounted articulated on the end of the guiding arm about an axis perpendicular to the part 12 . this hinge 22 is controlled by a cylinder 23 , one end of which carries the piston 24 and is fixed to the part 12 at the level of the joint 22 , with the other end carrying the cylinder 25 and being mounted for pivoting on the end of the guiding arm 9 . then , the cylinder 22 enables the passage of the guiding arm from the horizontal position with respect to the plane of the operating table 2 , in which it is kept through the blocking of the bearing lug 21 , to a downward sloping position . advantageously , the cylinder 23 is so arranged as to enable the stopping of the guiding arm with respect to the plane of the operating table 2 included in the solution provided between 0 degree and − 90 degrees . it should be noted that this chosen angular motion of the guiding arm results from the configuration of the implemented fastening means and the principle of the fastening selected for the guiding arm on the operating table . advantageously , the system lifting up the handle bar can be slowed in order to improve the utilisation ergonomics . advantageously , the cylinder 23 can be a simple gas cylinder slowly returning the guiding arm 9 as soon as the lowering effort on the latter has ceased , or a blocking gas cylinder in which the guiding arms 9 and 10 are kept in the sloping position in which they have been left . according to a particular aspect of the invention , the operating table 2 is motor - driven . the guiding device will advantageously comprise a control block 26 making it possible to control the motion of said operating table 2 , with the control arms 9 and 10 extending on either side of this control block 26 . for this purpose , the control block 26 is coupled to a base positioned under the table 2 and comprises an engine block . said base is a part of the frame supporting the top 3 . the words “ controlling the motion of the operating table ” means the starting and the stopping of the motor , the control of the front operation or rear operation of the table as well as the control of the speed motion of said table 2 . these actions are obtained by means of specific control buttons 27 , 28 and 29 . advantageously , the control block 26 is associated with the remote control device 30 making it possible to control the position of the operating table 2 and more particularly to adjust the positions of the various parts composing the top 3 of the operating table 2 . according to a specific configuration of the invention , said control block 26 includes a housing so configured as to removably receive said remote control device 30 . said housing is provided with connections matching the connections of the remote control device . advantageously , said remote control device 30 can be interconnected and is interchangeable . the guiding device 1 including a control block 26 and the remote control device 30 has a balanced shape to prevent the rotation motion observed on the systems of the prior art when they are dismounted from the operating tables . the invention has been described hereabove as an example . of course , the persons skilled in the art will have the possibility of making various modifications without leaving the scope of the invention .

the invention concerns a guiding device for a mobile operating table comprising at least one lateral rail for accessories , the aforementioned device comprising at least one guiding arm stretching along an axis and presenting an extremity fitted with a fastening mechanism permitting a detachable fixing of the aforementioned guide arm on the rail of the operating table , characterized in that the fastening means consist of at least one notch oriented along an axis substantially perpendicular to the axis of the guiding arm , said notch being capable of accommodating a spacer linking the side rail to the operating table , and a blocking element configured to block the guide arm on the rail by pressure of the aforementioned blocking element .

preferred embodiments of the present invention includes ( but not limited to ) the following : 1 . a prosthetic bone implant comprising a cortical portion having two opposite sides , and a cancellous portion integrally disposed in said cortical portion and being exposed through said two opposite sides , wherein said cortical portion comprises a hardened calcium phosphate cement has a porosity of less than 40 % in volume , and said cancellous portion comprises a porous hardened calcium phosphate cement having a porosity greater than 20 % in volume , and greater than that of said cortical portion . 2 . the implant according to item 1 , wherein the cortical portion is in the form of a hollow disk , and the cancellous portion is in the form of a column surrounded by the hollow disk . 3 . the implant according to item 2 further comprising a transitional portion between said column and said hollow disk surrounding said central cylinder , which has properties range from those of said cancellous portion to said cortical portion . 4 . the implant according to item 1 , wherein the cortical portion is in the form of a disk having one or more longitudinal through holes , and the cancellous portion is in the form of one or more columns surrounded by said one or more longitudinal through holes . 5 . the implant according to item 1 , wherein said hardened calcium phosphate cement of said cortical portion comprises an apatitic phase as a major phase giving rise to broadened characteristic x - ray diffraction peaks in comparison with a high - temperature sintered apatitic phase . 6 . the implant according to item 5 , wherein said broadened characteristic the x - ray diffraction peaks are at 2 - theta values of 25 – 27 ° and 30 – 35 °. 7 . the implant according to item 1 , wherein said hardened calcium phosphate cement of said cortical portion is prepared without a high temperature sintering . 8 . the implant according to item 1 , wherein said hardened calcium phosphate cement of said cortical portion comprises an apatitic phase as a major phase having a ca / p molar ratio of 1 . 5 – 2 . 0 . 9 . the implant according to item 1 , wherein said hardened calcium phosphate cement of said cancellous portion comprises an apatitic phase as a major phase giving rise to broadened characteristic x - ray diffraction peaks in comparison with a high - temperature sintered apatitic phase . 10 . the implant according to item 9 , wherein said broadened characteristic the x - ray diffraction peaks are at 2 - theta values of 25 – 27 ° and 30 – 35 °. 11 . the implant according to item 1 , wherein said hardened calcium phosphate cement of said cancellous portion is prepared without a high temperature sintering . 12 . the implant according to item 1 , wherein said hardened calcium phosphate cement of said cancellous portion comprises an apatitic phase as a major phase having a ca / p molar ratio of 1 . 5 – 2 . 0 . 13 . the implant according to item 1 , wherein said cortical portion comprises 10 – 90 % in volume of said implant . 14 . the implant according to item 1 , wherein said cortical portion has a porosity of less than 30 % in volume . 15 . the implant according to item 1 , wherein said cancellous portion has a porosity greater than 40 – 90 % in volume . 16 . a method for preparing a prosthetic bone implant comprising a cortical portion having two opposite sides , and a cancellous portion integrally disposed in said cortical portion and being exposed through said two opposite sides , said method comprises the following steps : a ) preparing a first paste comprising a first calcium phosphate cement and a first setting liquid ; b ) preparing a second paste comprising a second calcium phosphate cement , a pore - forming powder and a second setting liquid ; c ) i ) preparing a shaped article in a mold having two or more cells separated by one more partition walls comprising introducing said first paste and said second paste into said two or more cells separately , and removing said one or more partition walls from said mold , so that said second paste in the form of a single column or two or more isolated columns is integrally disposed in the first paste in said mold ; or ii ) preparing a shaped article comprising introducing one of said first paste and said second paste into a first mold to form an intermediate in said first mold , placing said intermediate into a second mold after a hardening reaction thereof undergoes at least partially , and introducing another one of said first paste ad said second paste into said second mold , so that said second paste as a single column or as two or more isolated columns is integrally disposed in the first paste in said second mold ; d ) immersing the resulting shaped article from step c ) in an immersing liquid for a first period of time so that said pore - forming powder is dissolved in the immersing liquid , creating pores in said single column or said two or more isolated columns ; and e ) removing the immersed shaped article from said immersing liquid . 18 . the method according to item 16 , wherein said pore - forming powder is selected from the group consisting of licl , kcl , nacl , mgcl 2 , cacl 2 , naio 3 , ki , na 3 po 4 , k 3 po 4 , na 2 co 3 , amino acid - sodium salt , amino acid - potassium salt , glucose , polysaccharide , fatty acid - sodium salt , fatty acid - potassium salt , potassium bitartrate ( khc 4 h 4 o 6 ), potassium carbonate , potassium gluconate ( kc 6 h 11 o 7 ), potassium - sodium tartrate ( knac 4 h 4 o 6 . 4h 2 o ), potassium sulfate ( k 2 so 4 ), sodium sulfate , and sodium lactate . 19 . the method according to item 16 , wherein said first calcium phosphate cement comprises at least one ca source and at least one p source , or at least one calcium phosphate source ; and said second calcium phosphate cement comprises at least one ca source and at least one p source , or at least one calcium phosphate source . 20 . the method according to item 19 , wherein said first calcium phosphate cement comprises at least one calcium phosphate source , and said second calcium phosphate cement comprises at least one calcium phosphate source . 21 . the method according to item 20 , wherein said calcium phosphate source is selected from the group consisting of alpha - tricalcium phosphate ( β - tcp ), beta - tricalcium phosphate ( β - tcp ), tetracalcium phosphate ( ttcp ), monocalcium phosphate monohydrate ( mcpm ), monocalcium phosphate anhydrous ( mcpa ), dicalcium phosphate dihydrate ( dcpd ), dicalcium phosphate anhydrous ( dcpa ), octacalcium phosphate ( ocp ), calcium dihydrogen phosphate , calcium dihydrogen phosphate hydrate , acid calcium pyrophosphate , anhydrous calcium hydrogen phosphate , calcium hydrogen phosphate hydrate , calcium pyrophosphate , calcium triphosphate , calcium phosphate tribasic , calcium polyphosphate , calcium metaphosphate , anhydrous tricalcium phosphate , tricalcium phosphate hydrate , and amorphous calcium phosphate . 22 . the method according to item 21 , wherein said first calcium phosphate cement and said second calcium phosphate cement are identical . 23 . the method according to item 22 , wherein said first calcium phosphate cement and said second calcium phosphate cement are tetracalcium phosphate . 24 . the method according to item 16 , wherein the first setting liquid and the second setting liquid independently are an acidic solution , a basic solution , or a substantially pure water . 25 . the method according to item 24 , wherein said acidic solution is selected from the group consisting of nitric acid ( hno 3 ), hydrochloric acid ( hcl ), phosphoric acid ( h 3 po 4 ), carbonic acid ( h 2 co 3 ), sodium dihydrogen phosphate ( nah 2 po 4 ), sodium dihydrogen phosphate monohydrate ( nah 2 po 4 . h 2 o ), sodium dihydrogen phosphate dihydrate , sodium dihydrogen phosphate dehydrate , potassium dihydrogen phosphate ( kh 2 po 4 ), ammonium dihydrogen phosphate ( nh 4 h 2 po 4 ), malic acid , acetic acid , lactic acid , citric acid , malonic acid , succinic acid , glutaric acid , tartaric acid , oxalic acid and their mixture . 26 . the method according to item 22 , wherein said basic solution is selected from the group consisting of ammonia , ammonium hydroxide , alkali metal hydroxide , alkali earth hydroxide , disodium hydrogen phosphate ( na 2 hpo 4 ), disodium hydrogen phosphate dodecahydrate , disodium hydrogen phosphate heptahydrate , sodium phosphate dodecahydrate ( na 3 po 4 . 12h 2 o ), dipotassium hydrogen phosphate ( k 2 hpo 4 ), potassium hydrogen phosphate trihydrate ( k 2 hpo 4 . 3h 2 o ), potassium phosphate tribasic ( k 3 po 4 ), diammonium hydrogen phosphate (( nh 4 ) 2 hpo 4 ), ammonium phosphate trihydrate (( nh 4 ) 3 po 4 . 3h 2 o ), sodium hydrogen carbonate ( nahco 3 ), sodium carbonate na 2 co 3 , and their mixture . 27 . the method according to item 16 , wherein step c - i ) further comprises allowing said first paste and said second paste undergoing a hardening reaction in said mold . 28 . the method according to item 16 , wherein step c - i ) further comprises pressurizing said first paste and said second paste in said mold after removing said one or more partition walls from said mold to remove a portion of liquid from said first paste and said second paste , so that a liquid / powder ratio of said first paste and of said second paste decreases ; and allowing said first paste and second paste undergoing a hardening reaction in said mold . 29 . the method according to item 16 , wherein step c - ii ) further comprises allowing said intermediate undergoing a hardening reaction in said first mold , and allowing said another one of said first paste and said second paste undergoing a hardening reaction in said second mold . 30 . the method according to item 16 , wherein step c - ii ) further comprises pressurizing said one of said first paste and said second paste in said first mold to remove a portion of liquid therefrom before the hardening reaction of said intermediate is completed ; allowing said intermediate undergoing a hardening reaction in said first mold ; pressuring said another one of said first paste and said second paste in said second mold , so that a liquid / powder ratio of said another one of said first paste and of said second paste decreases ; and allowing said another one of said first paste and second paste undergoing a hardening reaction in said second mold . 31 . the method according to item 28 , wherein said pressuring is about 1 to 500 mpa . 32 . the method according to item 30 , wherein said pressuring is about 1 to 500 mpa . 33 . the method according to item 16 , wherein the immersing liquid is an acidic aqueous solution , a basic aqueous solution , a physiological solution , an organic solvent , or a substantially pure water . 34 . the method according to item 33 , wherein the immersing liquid comprises at least one of ca and p sources . 35 . the method according to item 33 , wherein the immersing liquid is a hanks &# 39 ; solution , a hcl aqueous solution or an aqueous solution of ( nh 4 ) 2 hpo 4 . 36 . the method according to item 16 , wherein the immersing in step d ) is carried out for a period longer than 10 minutes . 37 . the method according to item 16 , wherein the immersing in step d ) is carried out for a period longer than 1 day . 38 . the method according to item 16 , wherein the immersing in step d ) is carried out at a temperature of about 10 and 90 ° c . 39 . the method according to item 38 , wherein the immersing in step d ) is carried out at room temperature . 40 . the method according item 17 further comprising cleaning said immersed shaped article before said drying ; and heating the resulting dried shaped article at a temperature between 50 and 500 ° c . four different designs of prosthetic bone implants constructed according to the present invention are shown in fig1 a to 1 d . in fig1 a , the prosthetic bone implant of the present invention has a dense cortical portion d 1 in the tubular form and a porous cancellous portion p 1 formed in the central through hole of the tubular cortical portion d 1 . both the dense cortical portion d 1 and the porous cancellous portion p 1 are made of a hardened calcium phosphate cement having an apatitic phase as a major phase . in fig1 b , the prosthetic bone implant of the present invention has a dense cortical portion d 1 in the tubular form ; a cylindrical porous cancellous portion p 1 in the center of the tubular cortical portion d 1 ; and an annular transitional portion p 2 connecting the tubular cortical portion d 1 and the cylindrical cancellous portion p 1 . the transitional portion p 2 is made of a hardened calcium phosphate cement having an apatitic phase as a major phase , and a porosity gradient increasing from the lower porosity of the cylindrical cancellous portion p 1 to the higher porosity of the tubular cortical portion d 1 , which may be formed in - situ during molding of two different two different cpc pastes , one of them having an additional pore - forming powder for forming the cylindrical cancellous portion p 1 , and another one being a regular cpc powder for forming the dense cortical portion d 1 . the porous cancellous portion p 1 may be in the forms of isolated columns surrounded by the dense cortical portion d 1 as shown in fig1 c and 1 d . other designs are also possible in addition to those shown in fig1 a to 1 d . a suitable method for preparing the prosthetic bone implant of the present invention includes placing a tubular partition wall 10 in a hollow cylindrical mold 20 , as shown in fig2 a ; pouring a first paste comprising a calcium phosphate cement and a setting liquid in the annular cell and a second paste comprising the calcium phosphate cement , a pore - forming powder and the setting liquid in the central cell , as shown in fig2 b ; removing the partition wall and pressing the cpc pastes before hardening , as shown in fig2 c , wherein a portion of the setting liquid is removed from the gap between the mold 20 and the press 30 and / or holes ( not shown in the drawing ) provided on the press 30 . the cpc paste will undergo a hardening reaction to convert into apatitic phase . the hardened disk is removed from the mold and is subjected to surface finishing to expose the central portion hardened from the second paste , as shown in fig2 d , followed by immersing in a bath of an immersing liquid as shown in fig2 e , where the pore - forming powder is dissolved in the immersing liquid while the hardened cpc thereof gaining compressive strength . the immersing may last from 10 minutes to several days . the composite disk so formed is washed with water after being removed from the bath , and dried and heated in an oven to obtain the prosthetic bone implant as shown in fig2 f . the heating is conducted at a temperature between 50 and 500 ° c . for a period of several hours to several days , which enhance the compressive strength of the cortical portion of the prosthetic bone implant . an alternative method for preparing the prosthetic bone implant of the present invention from the same raw materials includes pouring the second paste in a first mold , pressing the second paste to remove a portion of the setting liquid from the second paste before the hardening reaction is completed , so that the liquid / powder ratio in the second paste decreases , and allowing the hardening reaction undergo in the mold for a period of time , e . g . 15 minutes starting from the mixing of the cpc powder , the pore - forming powder and the setting liquid , to obtain a cylindrical block having a diameter of 7 mm . then , the cylindrical block is removed from the first mold , and placed in the center of a second mold having a diameter of 10 mm . the first paste is poured into the annular space in the second mold , and a press having a dimension corresponding to the annular shape is used to pressure the first paste to remove a portion of the setting liquid from the first paste before the hardening reaction is completed , so that the liquid / powder ratio in the first paste decreases . again , the first paste will undergo a hardening reaction to convert into apatitic phase . the hardened cylinder having a diameter of 10 mm is removed from the second mold , followed by immersing in an immersing liquid , where the pore - forming powder contained in the second paste is dissolved in the immersing liquid while the hardened cpc thereof gaining compressive strength , to obtain the prosthetic bone implant of the present invention , as shown in fig3 a and 3 b . it is apparently to people skilled in the art that the prosthetic bone implant shown in fig3 a and 3 b can also be prepared by changing the sequence of the molding of the first paste and the second paste with modifications to the second mold used in this alternative method . the following examples are intended to demonstrate the invention more fully without acting as a limitation upon its scope , since numerous modifications and variations will be apparent to those skilled in this art . a ca 4 ( po 4 ) 2 o ( ttcp ) powder was prepared by mixing ca 2 p 2 o 7 powder with caco 3 powder uniformly in ethanol for 24 hours followed by heating to dry . the mixing ratio of ca 2 p 2 o 7 powder to caco 3 powder was 1 : 1 . 27 ( weight ratio ) and the powder mixture was heated to 1400 ° c . to allow two powders to react to form ttcp . the resulting ttcp powder from preparative example 1 was sieved and blended with dried cahpo 4 ( dcpa ) powder in a ball mill for 12 hours . the blending ratio of the ttcp powder to the dcpa powder was 1 : 1 ( molar ratio ) to obtain the conventional cpc powder . particles of this c - cpc powder have no whisker on the surfaces thereof . the ttcp powder prepared according to the method of preparative example 1 was sieved and blended with dried cahpo 4 ( dcpa ) powder in a ball mill for 12 hours . the blending ratio of the ttcp powder to the dcpa powder was 1 : 1 ( molar ratio ). the resultant powder mixture was added to a 25 mm diluted solution of phosphate to obtain a powder / solution mixture having a concentration of 3 g powder mixture per 1 ml solution while stirring . the resulting powder / solution mixture was formed into pellets , and the pellets were heated in an oven at 50 ° c . for 10 minutes . the pellets were then uniformly ground in a mechanical mill for 20 minutes to obtain the non - dispersive ttcp / dcpa - based cpc powder ( nd - cpc ). the particles of this nd - cpc powder have whisker on the surfaces thereof . to a setting solution of 1m phosphoric acid solution ( ph = 5 . 89 ) the nd - cpc powder from preparative example 3 was added in a liquid / powder ratio ( l / p ratio ) of 0 . 4 , i . e . 4 ml liquid / 10 g powder , while stirring . the resulting paste was filled into a cylindrical steel mold having a length of 12 mm and a diameter of 6 mm , and was compressed with a gradually increased pressure until a maximum pressure was reached . the maximum pressure was maintained for one minute , and then the compressed cpc block was removed from the mold . at the 15 th minute following the mixing of the liquid and powder , the compressed cpc block was immersed in a hanks &# 39 ; solution for 1 day , 4 days , and 16 days . each test group of the three different periods of immersion time has five specimens , the compressive strength of which was measured by using a ags - 500d mechanical tester ( shimadzu co ., ltd ., kyoto , japan ) immediately following the removal thereof from the hanks &# 39 ; solution without drying . the cpc paste in the mold was compressed with a maximum pressure of 166 . 6 mpa , and in the course of the compression the compression speeds were about 5 mm / min during 0 ˜ 104 . 1 mpa ; 3 mm / min during 104 . 1 ˜ 138 . 8 mpa ; 1 mm / min during 138 . 8 ˜ 159 . 6 mpa : and 0 . 5 mm / min during 159 . 6 ˜ 166 . 6 mpa . the measured wet specimen compressive strength is listed table 1 . * this value was measured before the compressed cpc blocks were immersed in the hanks &# 39 ; solution , and it was substantially the same for the compressed cpc blocks not immersed in the hanks &# 39 ; solution measured a few days after the preparation . it can seen from table 1 that the compressive strength of the compressed cpc blocks is increased remarkably after one - day immersion in comparison with the non - immersed block , and declines a little for a longer immersion time . the procedures of example 1 were repeated by using the c - cpc powder prepared in preparative example 2 and the nd - cpc powder prepared in preparative example 3 . the maximum pressure used to compress the cpc paste in the mold in this example was 156 . 2 mpa . the results for one - day immersion time are listed in table 2 . it can be seen from table 2 that the compressive strength , 62 . 3 mpa , of the immersed compressed cpc block prepared from the conventional cpc powder ( no whisker ) is about 1 . 7 times of that ( 37 . 3 mpa ) of the non - immersed compressed cpc block in table 1 , and the compressive strength , 138 . 0 mpa , of the immersed compressed cpc block prepared from the non - dispersive cpc powder ( with whisker ) is about 3 . 7 times of that of the non - immersed compressed cpc block in table 1 ca 4 ( po 4 ) 2 o ( ttcp ) powder as synthesized in preparative example 1 was sieved with a # 325 mesh . the sieved powder has an average particle size of about 10 μm . to the ttcp powder hcl aqueous solution ( ph = 0 . 8 ) was added according to the ratio of 1 g ttcp / 13 ml solution . the ttcp powder was immersed in the hcl aqueous solution for 12 hours , filtered rapidly and washed with deionized water , and filtered rapidly with a vacuum pump again . the resulting powder cake was dried in an oven at 50 ° c . the dried powder was divided into halves , ground for 20 minutes and 120 minutes separately , and combined to obtain the non - dispersive ttcp - based cpc powder , the particles of which have whisker on the surfaces thereof . a setting solution of diammonium hydrogen phosphate was prepared by dissolving 20 g of diammonium hydrogen phosphate , ( nh 4 ) 2 hpo 4 , in 40 ml deionized water . the procedures in example 1 were used to obtain the wet specimen compressive strength for one - day immersion time , wherein the maximum pressure to compress the cpc paste in the mold was 156 . 2 mpa . the results are shown in table 3 . the procedures of example 1 were repeated except that the maximum pressure used to compress the cpc paste in the mold was changed from 166 . 6 mpa to the values listed in table 4 . the period of immersion was one day . the results are listed in table 4 . the data in table 4 indicate that the compressive strength of the cpc block increases as the pressure used to compress the cpc paste in the mold increases . effect of reducing liquid / powder ratio during compression of the cpc paste in the mold on compressive strength of nd - cpc block the procedures of example 1 were repeated except that the maximum pressure used to compress the cpc paste in the mold was changed from 166 . 6 mpa to the values listed in table 5 . the liquid leaked from the mold during compression was measured , and the liquid / powder ratio was re - calculated as shown in table 5 . the period of immersion was one day . the results are listed in table 5 . the procedures of example 1 were repeated . the period of immersion was one day . the cpc blocks after removing from the hanks &# 39 ; solution were subjected to post - heat treatments : 1 ) 50 ° c . for one day ; and 2 ) 400 ° c . for two hours with a heating rate of 10 ° c . per minute . the results are listed in table 6 . to a setting solution of 1m phosphoric acid solution ( ph = 5 . 89 ) the nd - cpc powder from preparative example 3 was added in a liquid / powder ratio ( l / p ratio ) of 0 . 4 , i . e . 4 ml liquid / 10 g powder , while stirring . kcl powder in a predetermined amount was mixed to the resulting mixture by stirring intensively . the resulting paste was filled into a cylindrical steel mold having a length of 12 mm and a diameter of 6 mm , and was compressed with a gradually increased pressure until a maximum pressure of 3 . 5 mpa was reached . the maximum pressure was maintained for one minute , and then the compressed cpc block was removed from the mold . at the 15 th minute following the mixing of the liquid and powders , the compressed cpc block was immersed in a deionized water at 37 ° c . for 4 days , 8 days , and 16 days . the compressive strength of the specimens of the three different periods of immersion time was measured by using a ags - 500d mechanical tester ( shimadzu co ., ltd ., kyoto , japan ) after the specimens were dry . the measured dry specimen compressive strength is listed table 7 . the procedures of example 7 were repeated by using sugar , ki , c 17 h 33 coona and c 13 h 27 cooh instead of kcl . the immersion time was 14 days in deionized water . in the cases where the c 17 h 33 coona and c 13 h 27 cooh were used , the cpc blocks were further immersed in ethanol for additional four days . the conditions and the results are listed in table 8 . it can be seen from table 8 that various powders which are soluble in water can be used in the preparation of a porous cpc block according to the method of the present invention . to a setting solution of 1m phosphoric acid solution ( ph = 5 . 89 ) the nd - cpc powder from preparative example 3 was added in a liquid / powder ratio ( l / p ratio ) of 0 . 4 , i . e . 4 ml liquid / 10 g powder , while stirring . kcl powder in a ratio of kcl powder / cpc by volume of 2 was mixed to the resulting mixture by stirring intensively . the resulting paste was filled into a cylindrical steel mold having a length of 12 mm and a diameter of 7 mm , and was compressed with a gradually increased pressure until a maximum pressure of 3 . 5 mpa was reached . the maximum pressure was maintained for one minute , and then the compressed cpc block was removed from the mold at the 15 th minute following the mixing of the liquid and powders . the resulting cylinder having a diameter of 7 mm was placed in another cylindrical steel mold having a diameter of 10 mm . to a setting solution of 1m phosphoric acid solution ( ph = 5 . 89 ) the nd - cpc powder from preparative example 3 was added in a liquid / powder ratio ( l / p ratio ) of 0 . 4 , i . e . 4 ml liquid / 10 g powder , while stirring . the resulting paste was filled into the gap between said cylinder and said another mold , and was compressed with a gradually increased pressure until a maximum pressure of 50 mpa was reached . the maximum pressure was maintained for one minute . at the 15 th minute following the mixing of the liquid and nd - cpc powder , the cpc / kcl composite block was immersed in a deionized water at 37 ° c . for 4 days . kcl powder was dissolved in the deionized water , and a dual - functional cpc block having a porous cpc cylinder surround by a dense cpc annular block was obtained . the compressive strength of the specimen was measured by using a ags - 500d mechanical tester ( shimadzu co ., ltd ., kyoto , japan ) after the specimens were dry . the measured dry specimen compressive strength is 68 . 8 mpa . the porosities of the porous cpc cylinder and the dense cpc annular block were measured by archimedes &# 39 ; method , and calculated as in astm c830 , after the dual - functional cpc block was broken intentionally , and the results are 74 % and 30 %, respectively . x - ray diffraction pattern of the powder obtained by grinding the dual - functional cpc block shows broadened characteristic x - ray diffraction peaks of apatite at 2θ = 25 – 27 ° and 2θ = 20 – 35 ° with a scanning range of 2θ of 20 – 40 ° and a scanning rate of 1 °/ min . results indicate that the powder is a mixture of apatite and ttcp with apatite as a major portion .

the present invention discloses a prosthetic bone implant made of a hardened calcium phosphate cement having an apatitic phase as a major phase , which includes a dense cortical portion bearing the majority of load and a porous cancellous portion allowing a rapid blood / body fluid penetration and tissue ingrowth .

the steps of the process begin with the grinding of a press cake that has been formed in a press from roasted cocoa beans . while use of a press cake of cocoa is preferred as the starting material , it is obvious that other cocoa forms such as nibs may also be used . however , regardless of the physical form at this stage it is subsequently pulverized and formed into a homogeneous mass . the press cake is then ground in a conventional grinder and pulverized into a coarse , free - flowing powder . sufficient size reduction is attained that the resultant particles will be easily workable and will pass through the die . a preferred particle size is one in which the particles have a diameter of about one - eighth inch or less . thus , it should be understood that the particles need only be ground so that they will subsequently pass through the die without plugging the opening . it is usually preferred to effect grinding until the particles will pass through a suitable screen . as to the screen through which the particles are passed , it is preferably of such size to provide particles having a size of about 50 to 150 mesh . the resulting powder is then passed through a magnet grate to remove any small metal particles that may have escaped previous cleaning steps . the ground cocoa material is then stored until mixed with alkali solution . the preferred fat content of the cocoa is about from 0 to 16 weight %. an aqueous alkaline solution is prepared in accordance with accepted practice using compounds of alkali and alkaline earth metals . a suitable alkali compound , such as sodium hydroxide , sodium carbonate ( soda ash ), sodium bicarbonate ( nahco 3 ), potassium carbonate , potassium hydroxide , potassium bicarbonate , ammonium carbonate , ammonium bicarbonate , ammonium hydroxide , ammonia gas , magnesium carbonate , magnesium oxide , and mixtures thereof may be used . the alkaline solution should have a concentration of about 3 to 12 weight percent , and preferably about 5 to 12 weight percent . the solution is well mixed and then diluted as necessary with water . dilution should be sufficient to provide a maximum of 3 % anhydrous k 2 co 3 or its alkaline equivalent in the final product . with low fat nibs , up to 6 percent alkali can be present . supplemental colors , sugars , malts or the like may also be added at this stage . the alkaline solution additive is then mixed with the proper proportion of powdered cocoa material and good mixing achieved . at this stage the mixture shoud have a moisture content of about 20 - 35 weight percent . the resulting damp mass or mixture is then subjected to continuous mechanical working . mixing may be effected using one or more rotating shafts with mullers , paddles and ribbons mounted on the shafts for thoroughly mixing the product mixture into a homogeneous state . this is particularly advantageous since upon next entering the cooking step , the cooking may thus be performed in a uniform manner without creating hot spots or uncooked portions that might otherwise result due to a non - homogeneous mixture . a suitable piece of equipment to perform this step is a &# 34 ; double shaft continuous mixer &# 34 ; manufactured by paul o . abbe , inc . of little falls , new jersey . while this is the preferred manner of carrying out this step of the process it is to be understood that any type of mechanical working can be utilized , it being only necessary that the mixture be subjected to such thorough mixing that a homogeneous mass will be provided prior to the cooking step . the product mixture is now ready to enter the next important step of the process of the present invention . this step continues the mechanical working of the product mixture and concurrently cooks the mixture under pressure that progressively increases . the preferred equipment used is the cooking extruder manufactured by the bonnot company of kent , ohio , although it is to be understood that equivalent equipment may be utilized in accordance with the broad aspects of the invention . in the preferred equipment , the increase in pressure is provided by a variable pitch screw . it should be understood that other types of mechanical working may be used within the extruder . suffice it to say that the product mixture is continuously worked by the screw and is progressively pressurized while at the same time being heated to cook the product . the exit end of the extruder is provided with an extruding die head from which the compressed product mass is extruded . the extruding step forms a product rod that is divided into relatively short lengths to form product pellets . in the cooker - extruder , the temperature is maintained at about 150 ° to 230 ° f ., preferably about 180 ° to 212 ° f ., while progressively increasing the pressure to a final stage of pressurization of about 500 to 1300 pounds per square inch , preferably about 700 to 1100 psig . in general , the homogeneous mixture is heated substantially by the frictional forces or mechanical working taking place within the extruder which can operate adiabatically but additional heating means is provided if it is necessary to maintain the indicated temperature range . temperatures above those indicated may result in charring of the product . the residence or cooking time within the cooker - extruder will vary over a limited range with limitations being placed thereon by the size of the die opening and the speed of the screw . suitable die openings range from approximately 0 . 100 inch to about 1 . 150 inch . larger opening may be used but the product quality and economies of operations may be adversely affected . much larger openings would also be unsuitable because the required pressures could not be produced . in general , it may be stated that in operation of a continuous process with continuous introduction of cocoa material , for a die having a diameter ranging from about 0 . 100 inch to about 0 . 150 inch , the cooking or residence time can range from about two and one - half nimutes to about five minutes . a preferred residence or cooking time within the cooker - extruder is about four to five minutes . the time within the cooker is important , the time within the die being only about one second or less . the head of the extruder includes any number of suitable extruding dies or passageway out of which come the lengths of product rod . the composition of the product is gauged by moisture content of about 20 - 35 weight percent , preferably about 27 - 33 percent , in order to facilitate breaking off of the rod cleanly as it leaves the extruder head . this breaking off process is facilitated by providing a stationary plate across the path of the exiting rod . the plate breaks the rod into pellets . at this stage the pellets may be used as desired or they may be dried by any desired process . a particular method of drying is disclosed herein . prior to drying , the pellets still contain about 85 - 97 percent of the initial water added to the press cake . in the preferred drying procedure the pellets are dried in a fluidized bed dryer and cooler unit wherein a bed of the pellets if formed by directing a plurality of downwardly extending streams of heated gas into a bed to lift and tumble the pellets . the gas is preferably an inert gas such as nitrogen or argon . the jet streams of gas are provided from a heating plenum and a cooling plenum or other desired source . the bed of pellets may be oscillated or vibrated in order to effect feeding along in a continuous fashion . at the exit end of the dryer , the dried and cooled pelletized end product is recovered for further processing and forwarding to the converter or maker of the final cocoa product . a particular fluidized bed dryer is sold as the &# 34 ; jetzone &# 34 ; and manufactured by wolverine corporation , methuen , massachusetts . other equivalent euipment may also be used of course . thus having described the equipment and the steps of the process , a more particular example of the process and the results can be set forth in the following example . particular attention should be directed in this example to the operating conditions placed on the equipment that greatly enhance the efficiency and desired results of the process . cocoa material in the press cake form was obtained having preferably 10 to 16 percent fat . the press cake was comminuted through a pulverizer or grinder so as to form the product into a coarse , free - flowing powder of sufficiently small size that it would not plug the one - eighth inch die opening . a particle size of about 60 mesh was suitable . the powder was metered into a mixer at a rate of approximately 972 grams per minute . a final diluted solution of 22 . 35 grams of anhydrous sodium ash ( na 2 co 3 ) in 390 grams of water was prepared and fed into the mixer at a predetermined rate to give a 33 percent moisture basis . the mixer was operated in a continuous fashion to completely combine and mix the soda ash solution with the powdered cocoa material , and the resultant homogeneous product mixture was then introduced into the cooking extruder at a continuous rate . temperatures within the extruder were controlled in the range at 190 ° f . and the pressures were progressively increased and in the final stage were controlled to 900 pounds per square inch . the product was thus continuously mechanically worked by an auger as the cooking was carried out . total time within the cooker - extruder was four and one - half minutes . the compressed product mass was continuously fed from the extruding head and formed into approximately one - eighth inch diameter rods as the mass was extruded through the one - eighth inch die holes . this rod contained approximately 29 percent moisture ; a loss of 4 percent moisture occurring as the rod emerges from the extruder 40 . this moisture content allowed the rod to be freely broken into the pellets by the presence of a stationary plate . the pellets formed were from 1 / 4 inch to 1 inch in length . because of the control of the moisture content and the equipment used , there was no need to employ a cutter in order to cut the rod . these pellets were particularly suitable for drying and cooling because of the large surface area exposed to the gas of the fluidized bed unit . the plenum of the fluidized bed unit was provided with a heated inert gas ( nitrogen ) that was injected into the bed of pellets in the drying section to lift and tumble the pellets for approximately 2 . 5 to 3 minutes as the bed moved continuously along a support . the heating element raised the temperature and maintained the same in the range of 350 ° to 450 ° f . finally , the pellets were cooled to about ambient temperature under the gas nozzles in about 2 . 5 to 3 minutes . the product had a rich dark color and a moisture content of about 4 weight percent . in summary , dutching of cocoa is performed by grinding the pressed cake made from cocoa beans , converting to powder , mixing with alkaline solution , and cooking the product mixture under pressure and continuous mechanical working . the compressed product of critical moisture is passed through die means to form pellets by breaking into relatively short lengths . the drying and cooling of the pellets is performed in an efficient manner on a fluidized bed with the pellets being lifted and tumbled by downwardly directed jet streams of inert gas . in the process of this application the initial moisture content , preliminary mixing to a homogeneous mass and residence time within the cooker - extruder are especially critical in order to provide a final cocoa product having a rich homogeneous color , improved flavor and high sterility . by use of the term &# 34 ; rich color &# 34 ; is meant a dark uniform color whether brown , red or black . in this disclosure , there is shown and described only the preferred embodiment of the invention , but , as aforementioned , it is to be understood that the invention is capable of use in various other combinations and environment and is capable of changes or modifications within the scope of the inventive concept as expressed herein .

the dutching of cocoa to obtain a product of rich homogeneous color and high sterility is accomplished by grinding a pressed cake made from cocoa beans , converting the ground mass into a coarse , free - flowing powder , mixing the powder with sufficient aqueous alkaline solution and desired additives to obtain a damp mass containing about 20 - 35 wt . per cent moisture and feeding this mixture into a combined cooker and pressurized extruder and then pelletizing the cooked composition . the pellets are formed by extruding the mass through appropriate dies and breaking off the rods to form pellets by interaction with a stationary plate . the preferable temperature range of the combined cooking - pressurizing step is 150 °- 230 ° f . and the preferred pressures are progressive to between 500 and 1300 pounds per square inch adjacent the extruding die . the pellets are optionally dried in a continuous fashion by a plurality of gas jet streams that lift and tumble the pellets continuously passing underneath on a vibrating conveyor .

an ocular solution containing an antioxidant that has been stabilized to retard deterioration of the antioxidant is disclosed . the ocular solution containing a stabilized antioxidant is able to participate in reactions that scavenge free radicals and thus preserve the physiological benefits of the antioxidant . in contrast , because antioxidants are inherently unstable and readily participate in free radical reactions during which they are auto - oxidized , an ocular solution containing an antioxidant that has not been stabilized is extremely susceptible to oxidation . this renders the antioxidant less active , and decreases , retards , or prevents the antioxidant from providing its beneficial properties to the ocular solution . the ocular solution to which the stabilized antioxidant is added may be any physiologically compatible ocular solution for use in any manner , either topically or invasively . it will be appreciated that the ocular solution containing the stabilized antioxidant need not be in the physical form of a true solution , but instead may be a suspension , a cream , an ointment , an emulsion , a gel , etc . thus , the term solution is used for convenience but encompasses other physical states in which the stabilized form of the antioxidant and the other components are present . it will also be appreciated that the stabilized antioxidant may be included in the formulation for preparing an ocular solution , or may be added in dry form or in the form of a concentrated solution to an already prepared ocular solution . the ocular solution may be one that is used as an ocular irrigating solution or as a volume replacement during ocular surgery . it may also be one that is used topically , and thus encompasses eye drops , eye wash solutions , and contact lens solutions . it may be used in over the counter ( otc ) ocular solutions for topical application , for example , in ocular solutions such as artificial tears or lubricants . one commercially available ophthalmic lubricant ( viva - drops ®, available from vision pharmaceuticals , inc . ( mitchell s . d .)) is reported to contain polysorbate 80 as an antioxidant active ingredient , and sodium citrate , citric acid , edta , retinyl palmitate , and sodium pyruvate as inactive ingredients and antioxidants . it may also be used in prescription ( rx ) ocular solutions for topical application . examples of prescription ophthalmic compositions include , but are not limited to , the following : loteprednol etabonate ophthalmic suspension 0 . 5 % ( lotemax ™) as an ophthalmic topical antiinflammatory corticosterioid ; loteprednol etabonate ophthalmic suspension 0 . 2 % ( alrex ™) as an ophthalmic topical anti - inflammatory cortocosteroid ; metipranolol ophthalmic solution 0 . 3 % ( optipranolol ™) as a non - selective beta - adrenergic receptor blocking agent ; sodium chloride 2 % or 5 % solution or ointment ( muro 128r ) as a treatment for corneal edema by drawing water out of the cornea of the eye , all available from bausch and lomb pharmaceuticals ( tampa fla . ); and trifluridine ophthalmic solution 1 % ( viropticr ) available from monarch pharmaceuticals , inc . ( bristol tenn .). the inventive composition may be used in physiologic ophthalmic irrigating solutions . one example is balanced salt solution ( bss ®, alcon laboratories ( randburg , south africa ), containing per ml 0 . 64 % sodium chloride , 0 . 075 % potassium chloride , 0 . 048 % calcium chloride , 0 . 03 % magnesium chloride , 0 . 39 % sodium acetate , and 0 . 17 % sodium citrate dihydrate , as well as sodium hydroxide and / or hydrochloric acid to adjust ph , and water for injection . another example is ocular irrigation solution ® ( allergan , irvine calif .). another example is lactated ringer &# 39 ; s solution . another example is a normal saline solution . another example is normal saline adjusted to ph 7 . 4 with sodium bicarbonate . the inventive composition may be used in ophthalmic volume replacement solutions for introduction into the posterior chamber of the eye to replace the vitreous that is removed during vitrectomy . as another example , it may be used as an ocular wash solution . any antioxidant in a physiological formulation for ocular administration may be used . one antioxidant is vitamin c , which is also known as ascorbic acid or l - ascorbic acid . vitamin c is unstable in the presence of oxygen and decomposes to form l - ascorbic acid 2 - hydrogen sulfate , and then dehydroascorbic acid . providing a stabilizing agent with vitamin c reduces or eliminates its tendency to be oxidized in solution , and hence the stabilizing agent guards against vitamin c deterioration . another example of an antioxidant is vitamin e ( α - tocopherol ). vitamin e may be in the form of tocopherol or its esters , for example , tocopheryl acetate . another example of an antioxidant is vitamin a , which may be in the form of retinol or its ester or acids , for example , retinyl palmitate or retinoic acid . thus , it will be appreciated that derivatives of vitamins c , e , and a are also included within the scope antioxidants . a stabilized form of any of these antioxidants may be used separately or in combination in the ocular solution . an antioxidant and a stabilizing agent for the antioxidant is included with ocular solutions for any use . the antioxidant and stabilizing agent may be added together or separately as individual components in the preparation of an ocular solution . alternatively , a solution of the antioxidant and stabilizing agent may be prepared and then added to the ocular solution . it will be appreciated that , if the ocular solution to which the antioxidant is to be added itself contains a stabilizing agent , then the separate addition of a stabilizing agent may be optional and the antioxidant may be added directly to the solution to result in a stabilized antioxidant . for example , some commercially available ocular solutions contain glutathione , which is an antioxidant stabilizing agent . the solutions may be commercial irrigating solutions that contain other known components , such as various anions and cations , buffers to regulate ph , adenosine , calcium , glucose , bicarbonate , dextrose , dextran 40 ( a low molecular weight colloidal osmotic agent ), gentamicin , dexamethasone , selenium , zinc , and gluconide . the antioxidant and stabilizing agent may be added to commercial ocular lubricating solutions , such as artificial tears . the antioxidant and stabilizing agent may be added to commercial ocular wash solutions . any solution for ocular administration , either administration to the exterior surface of the eye or to one of the interior chambers of the eye , may contain the antioxidant and stabilizer . in one embodiment , a sterile solution of the antioxidant such as vitamin c is prepared . a stabilized form of vitamin c is prepared by including in the vitamin c solution one or more components which inhibit , minimize , prevent , or decrease the extent of oxidation . one example of such a stabilizing component is cysteine . another example of such a component is l - cystine . another example is a solution of water up to about 12 % water and at least one organic solvent miscible with water , namely , ethanol , n - propanol , isopropanol , methanol , propylene glycol , butylene glycol , hexylene glycol , glycerine , sorbitol ( polyol ), di - propylene glycol , polypropylene glycol ( claim 1 of &# 39 ; 382 patent ), or a mixture of propylene glycol and butylene glycol with propylene glycol at about 25 % by weight to about 80 % by weight and butylene glycol at about 5 % by weight to about 30 % by weight and optionally including other glycols . another example is glutathione , such as reduced glutathione with selenium as a cofactor . another example is n - acetyl - l - cysteine . another example is l - methionine . another example is magnesium ions in at least 14 parts by weight to 100 parts by weight vitamin c . another example is a combination of at least one phosphonic acid derivative and at least one metabisulfite . another example is an antioxidant in a nonaqueous or substantially anhydrous silicone vehicle where the silicone vehicle comprises at least 50 % by weight of the composition . another example is acrylic and methacrylic polymers , or xanthans . another example is an extract of the fruit of the emblica officinalis plant , which contains , by weight , gallic / ellagic acid derivatives of 2 - keto - glucono - δ - lactone at about 35 % to about 55 %; punigluconin ( 2 , 3 - di - o - galloyl 4 , 6 -( s )- hexahydroxy - diphenoylgluconic acid at about 4 % to about 15 %; pedunculagin ( 2 , 3 , 4 , 6 - bis -( s )- hexahydroxydiphenoyl - d - glucose at about 10 % to about 20 %; rutin ( flavanol - 3 ) glycoside at about 5 % to about 15 %; low to medium molecular weight gallo - ellagi tannoids at about 10 % to about 30 %, gallic acid from 0 % to about 5 %, and ellagic acid from 0 % to about 5 %, available as capros from natreon inc . ( new brunswick n . j .). the antioxidant vitamins c , a , and e are available commercially from a number of sources ( e . g ., sigma - aldrich fine chemicals , st . louis mo .). a solution of vitamin c is prepared in a desired concentration . in one embodiment , the solvent is water . in another embodiment , the solvent is water and at least one organic liquid miscible with water . vitamin c derivatives may also be used , example of which include alkali salts such as sodium ascorbate and potassium ascorbate , alkaline earth salts such as calcium ascorbate and magnesium ascorbate , esters such as ascorbyl palmitate , ascorbyl laureate , ascorbyl myristate , ascorbyl stearate , other salts such as magnesium ascorbyl phosphate , ascorbyl - phosphoryl - cholesterol , dipalmitate ascorbate , and ascorbate anhydrides . the stabilizing agents are available commercially from a number of sources ( e . g ., sigma - aldrich ). cysteine and methionine are available as a hydrochloride salt or another physiologically acceptable salt , and may be added to the solution in amounts to yield an appropriate amount of the free base . embodiments of the invention include various concentrations of the antioxidants and stabilizer sufficient to stabilize the antioxidants against oxidation . concentrations of the antioxidant and stabilizer ( s ) may depend upon the use for the composition , as is known to one skilled in the art . thus , the invention is not limited to a specific concentration of either vitamin c or the stabilizing agent . in general , the antioxidant is present in the ocular solution at concentrations ranging from about 1 μg / ml to about 10 mg / ml . in embodiments , vitamin c , vitamin a , or vitamin e at concentrations in the range of about 0 . 025 mg / ml to about 1 . 2 mg / ml may be used , or concentrations in the range of about 0 . 1 mg / ml to 0 . 3 mg / ml may be used , or a concentration up to about 10 % of the final solution may be used , or a concentration in the range of about 10 % of the final solution to about 15 % of the final solution within the limits of solubility may be used . free cysteine is included at concentrations by weight of the vitamin of about 0 . 4 %, about 0 . 5 %, about 0 . 6 %, about 0 . 7 %, about 0 . 8 %, about 0 . 9 %, about 1 %, about 2 . 5 %, or about 5 % may be used , or within the range of about 0 . 2 % of the vitamin to about 2 . 3 % of the vitamin , or within the range of about 0 . 2 % of the vitamin to about 1 . 25 % of the vitamin , or within the range of about 0 . 3 % of the vitamin to about 0 . 9 % of the vitamin . in one embodiment , an ocular solution contains vitamin c , vitamin a , or vitamin e at a concentration in the range between about 1 % by weight to about 25 % by weight , glutathione in the range between about 0 . 01 % by weight to about 10 % by weight , a source of selenium as a cofactor for glutathione at a concentration in the range from about 0 . 001 % by weight to about 2 . 0 % by weight , and a sulfur - containing amino acid at a concentration in the range of about 0 . 001 % by weight to about 2 . 0 % by weight . in various embodiments , other precautions may also be taken to minimize or reduce oxidization and thus further enhance the stability of the ocular solution . for example , the ocular solution containing antioxidant may contain a chelating agent such as ethylenediamine tetraacedic acid ( edta ), it may be packaged under nitrogen , its exposure to light may be minimized , etc . the ocular solution may be an ocular wash solution , an ocular lubricating solution , an ocular irrigating solution , an ocular therapeutic solution , etc . the following references disclose methods which may be used in embodiments of the invention and are expressly incorporated by reference herein in their entirety : u . s . pat . nos . 3 , 958 , 017 ; 4 , 983 , 382 ; 5 , 281 , 196 ; 5 , 516 , 793 ; 5 , 703 , 122 ; 5 , 906 , 811 ; 6 , 804 , 110 ; 6 , 087 , 393 ; 6 , 103 , 267 ; 6 , 110 , 476 ; 6 , 146 , 664 ; 6 , 183 , 729 ; 6 , 211 , 231 ; 6 , 235 , 721 ; 6 , 299 , 889 ; 6 , 361 , 783 . the following examples are illustrative only , and do not limit the scope of the invention . an aqueous solution of up to about 10 % vitamin c , containing in the range of about 1 % cysteine to about 5 % cysteine is prepared . the stabilized vitamin c solution is prepared with or is incorporated into an ocular solution to achieve a final vitamin c concentration of about 0 . 1 % by weight to about 5 % by weight . an aqueous solution containing in the range of about 0 . 025 mg / ml vitamin c to about 1 . 2 mg / ml vitamin c , and in the range of about 1 % cysteine to about 5 % cysteine , is prepared . the stabilized vitamin c solution is prepared with or is incorporated into an ocular solution to achieve a final vitamin c concentration of about 0 . 1 % by weight to about 5 % by weight . an aqueous solution containing about 0 . 228 mg / ml vitamin c and in the range of about 1 % cysteine to about 5 % cysteine is prepared . the stabilized vitamin c solution is prepared with or is incorporated into an ocular solution to achieve a final vitamin c concentration of about 0 . 1 % by weight to about 5 % by weight . an aqueous solution containing about 0 . 1 mg / ml vitamin c to about 0 . 3 mg / ml vitamin c and in the range of about 1 % cysteine to about 5 % cysteine is prepared . the stabilized vitamin c solution is prepared with or is incorporated into an ocular solution to achieve a final vitamin c concentration of about 0 . 1 % by weight to about 5 . 0 % by weight . vitamin c at about 0 . 78 mg / ml ( 23 . 04 mg / ounce ) and free cysteine in the range of about 0 . 0097 mg / ml ( about 0 . 288 mg / 1 ounce ) to about 0 . 019 mg / ml ( about 0 . 576 mg / 1 ounce ) is prepared with or is incorporated into an ocular solution to achieve a final vitamin c concentration of about 0 . 1 % by weight to about 5 % by weight . vitamin c in the range between about 0 . 29 mg / ml to about 0 . 39 mg / ml , and cysteine hydrochloride anhydrous at a concentration of about 0 . 002 mg / ml as free cysteine , or in the range between about 0 . 505 % to about 0 . 685 % free cysteine by weight vitamin c , is prepared with or is added to an ocular solution . an ocular solution that may be a contact lens solution , a eye wash solution , an irrigating solution , a volume replacement solution , a therapeutic solution available either by prescription or over the counter , or a lubricant solution contains about 0 . 0340 % by weight vitamin c and 0 . 0002 % cysteine . an ocular solution containing up to about 10 % vitamin c and cysteine at a concentration in the range of about 0 . 2 % by weight of vitamin c to about 2 . 3 % by weight of the vitamin c is prepared . an ocular solution containing up to about 10 % vitamin c and cysteine at a concentration of about 0 . 588 % by weight vitamin c is prepared . an ocular solution containing in the range of about 0 . 0025 % vitamin c to about 0 . 12 % vitamin c , and cysteine at a concentration of about 0 . 588 % by weight vitamin c , is prepared . an ocular solution containing in the range of about 30 mg vitamin c to about 2000 mg vitamin c , and magnesium ions at least at 14 parts by weight in 100 parts of vitamin c , is blended at concentrations in the range of about 1 . 5 meq / liter to about 35 meq / liter . in 50 mm phosphate buffer ( ph 6 ), magnesium sulfate heptahydrate is dissolved at 2 . 054 g / liter , and vitamin c at 0 . 2 g / liter . the solution is transferred into polyethylene bags , replaced with nitrogen , and sterilized under nitrogen pressure for 15 min at 115 ° c . it is added to or formulated with an ocular solution to achieve a final vitamin c concentration in the range between about 0 . 1 % by weight to about 5 % by weight . to an ocular solution , the following components are added : 334 mg / liter vitamin c , 4 g / liter l - methionine , 1 . 1 g / liter n - acetyl - l - cysteine , and 2 . 054 g / liter magnesium sulfate heptahydrate . to an ocular solution , at least 5 % vitamin c and a mixture of propylene glycol and butylene glycol , with propylene glycol at about 25 % by weight to about 80 % by weight and butylene glycol at about 5 % by weight to about 30 % by weight , and optionally including other glycols , is added to an ocular solution , at least 5 % vitamin c and a mixture of propylene glycol and butylene glycol , with propylene glycol at about 25 % by weight to about 80 % by weight and butylene glycol at about 5 % by weight to about 30 % by weight , and optionally including other glycols , is added vitamin c is added to an ocular solution in an oil phase dispersion of particles . vitamin c , water , and a water soluble or water dispersible polymer ( s ) is prepared . the polymers may be natural or synthetic polymers , including but not limited to methacrylates , cellulosic polymers , polyethylene glycols and copolymers , natural or modified natural resins , polyvinyl resins , water - solubilized or water - dispersible polyurethanes , water - solubilized or water - dispersible ethers , etc . a solution of vitamin c ( in various embodiments , by weight of the dispersion / suspension , at least 5 %, at least 5 . 5 %, at least 6 %, at least 7 %, at least 8 . 5 %, at least 10 %, and up to 40 %, 50 %, 60 %, 75 %), water , and a water - soluble polymer is prepared and mixed with a solution of oil and water in a surface active agent having an hydrophilic - lipophilic balance of less than 12 . the solutions are dispersed to form a mixture , which is cooled to solidify the vitamin c containing solution to form particles dispersed in oil . emulsifying is performed at a temperature greater than 40 ° c . an ocular solution is prepared with vitamin c at a concentration in the range of about 0 . 01 % to about 20 %, at least one phosphonic acid derivative at a concentration between about 0 . 005 % to about 5 %, and at least one metabisulfite in a concentration between about 0 . 005 % to about 5 %. the phosphonic acid derivative may be ethylenediaminetetra ( methylenephosphonic acid ), hexamethylenediaminetetra ( methylenephosphonic acid ), diethylenetriaminepenta ( methylenephosphonic acid ), and their salts . the metabisulfite may be an alkali - metal , alkaline - earth , metal , or ammonium salt of anhydrosulfonic acid . the weight ratio between metabisulfite and phonphonic acid derivative ranges from 1 to 1000 . in one embodiment , the weight ratio ranges from 1 to 5 . vitamin c in a nonaqueous or substantially anhydrous silicone vehicle is prepared . in various embodiments , vitamin c is at a concentration ( all percentages are by weight ) of at least 0 . 1 %, at least 1 %, from about 2 % to about 30 %, from about 5 % to about 20 %, from about 8 % to about 12 %, or about 40 % of undissolved ascorbic acid . the carrier is an anydrous silicone carrier in an amount of about 50 % by weight to about 80 % by weight . the silicone vehicle may be a oil , gel , or solid . silicone includes organosiloxanes and polyorganosiloxanes . other antioxidants may be included . polysilicone - 11 ( from about 0 . 1 % to about 68 %), dimethicone ( from about 0 . 1 % to about 36 %), and cyclomethicone ( from about 0 . 1 % to about 56 %) are combined and the optional vitamins , if used , are added . solid vitamin c ( 10 %) is dispersed with agitation and is ground using a three - roll mill until the particle size is less than 20 μm and the mixture is uniform . in one embodiment , the particle size is less than 12 . 5 μm . in various embodiments , vitamin c is at a concentration ranging from about 5 % to about 70 %, from about 10 % to about 60 %, or from about 20 % to about 60 %. xanthan or acrylic and methacrylic polymers are added to a concentration ranging from about 0 . 1 % to about 5 %. the composition is prepared with or is incorporated in an ocular solution at a vitamin c concentration in the range between about 0 . 1 % to about 5 %. linoleic acid or an ester of linoleic acid may also be included . in various embodiments , vitamin c or derivatives of vitamin c and an extract of the fruit of the emblica officinalis plant are combined in a weight ratio of about 1 : 10 or about 10 : 1 . the use of the fruit extract of emblica officinalis as a stabilizer for vitamin c is described in u . s . pat . no . 6 , 235 , 721 which is expressly incorporated by reference herein in its entirety . the extract contains , by weight , ( 1 ) and ( 2 ) about 35 - 55 % of the gallic / ellagic acid derivatives of 2 - keto - glucono - δ - lactone ; ( 3 ) about 4 - 15 % of 2 , 3 - di - o - galloyl 4 , 6 -( s )- hexahydroxydiphenoyl - gluconic acid ; ( 4 ) about 10 - 20 % of 2 , 3 , 4 , 6 - bis -( s )- hexahydroxydiphenoyl - d - glucose ; ( 5 ) about 5 - 15 % of 3 ′, 4 ′, 5 , 7 - tetrahydroxyflavone - 3 - o - rhamnoglucoside ; and ( 6 ) about 10 - 30 % of tannoids of gallic / ellagic acid , gallic acid ( 0 - 5 %); ellagic acid ( 0 - 5 %) at a concentration ranging from about 5 % to about 70 %, from about 10 % to about 60 %, or from about 20 % to about 60 %. xanthan or acrylic and methacrylic polymers are added at a concentration ranging from about 0 . 1 % to about 5 %. the composition is incorporated in an ocular solution at a vitamin c concentration in the range between about 0 . 1 % to about 5 %. other variations or embodiments of the invention will also be apparent to one of ordinary skill in the art from the above figures and descriptions . thus , the forgoing embodiments are not to be construed as limiting the scope of this invention .

ocular solutions containing an antioxidant provide beneficial properties , for example , the antioxidant scavenges free radicals in the solution which may cause the solution to deteriorate . however , antioxidants are themselves extremely susceptible to oxidation . a stabilizing agent for the antioxidant retards or prevents the antioxidant from undesirable reactions and thus enhances its ability to stabilize the ocular solution . this in turn enhances the physiological properties of the ocular solution , which may be a topical solution such as eye drops , or a surgical ocular irrigation or volume replacement solution .

referring now to the drawings in greater detail , fig1 - 3 illustrate one embodiment 10 of the back and bail assembly of the present invention including the various portions thereof . the bail or handle portion 12 has a generally u - shaped configuration including generally parallel leg members 14 merging into enlarged heads 16 at the ends of the legs . each head portion 16 is generally cylindrical having a generally rectangular cross - sectional shape when viewed from the front ( fig1 ) and a circular cross - sectional shape when viewed from the side ( fig4 ). head portion 16 includes left and right end 18 and 20 ( fig1 ). on the outer end of each head portion 16 -- namely , the left end 18 on left leg 14 , and on the right end 20 on the right leg as shown in fig1 are included cylindrical pins or stud projections 22 extending to only one side of the leg , formed integrally with head 16 , and extending beyond the respective end surfaces . studs 22 generally coaxial with the head portion cylinders 16 and provide a pivot axis for the bail or handle 12 which is rotated therearound by grasping a flange 15 at the bottom of the bail when the bail is assembled with the back members . as shown generally in fig4 legs 14 lie generally in the same plane while the axes of cylindrical pins or stud projections 22 likewise lie in the same plane . as will be seen in fig1 studs or pins 22 extend in opposite directions to either lateral side of the bail or handle but only one pin is included on each leg . alternatively , pins 22 may extend toward one another , one from the inside of each leg . further , one pin 22 could be formed to extend from the inside of one leg while the other pin extends from the outside of the other leg . in any event , only a single pin need extend from each leg . on the end of each head portion 16 which is opposite from the end from which the pins or stud projections 22 extend , is a cut - away portion or area allowing the respective leg to be pivoted into a position between the supports of a back member . in the assembly embodiment shown in fig1 - 3 , the cut - away portion is formed on each head 16 by an inclined surface 24 recessed from the normal or regular outline of head 16 which provides a bevel across the bottom corner on the end opposite the pin or stud 22 -- namely , end 20 for the left leg and end 18 for the right leg ( fig1 ). surface 24 is inclined at an angle to the axis of cylindrical pin 22 . as is best seen in fig1 the bail or handle 12 is pivotally secured to a furniture member f by means of a pair of back members 30 , one back member being provided for each leg 14 . each back member includes a generally planar base portion 32 . projecting outwardly from one side of the base 32 , at spaced positions adjacent either end of the base , are a a pair of bail supports or flanges 34 . the inside wall surfaces 36 of the supports or flanges 34 are generally planar , extend generally perpendicular to base 32 , and oppose one another to define a pocket , recess , or slot therebetween for receiving the head 16 of one of the legs of the bail 12 . in order to receive the pin or stud projection 22 from one of the head portions , at least one of the wall surfaces 36 of back member 30 includes an elongated slot or recess 38 which extends outwardly along the wall surface away from base 32 and ends with a closed end 40 at a distance spaced from the base 32 . as seen in fig2 , and 5 , recess 38 extends through the base and opens to the back side of the base and also extends into the area of the base between the wall surfaces 36 and is therefore generally l - shaped in overall configuration . when assembled , the side surfaces of the pin or stud projections 22 abut against the ends 40 of the recesses 38 and pivot thereagainst such that the bail is pivotally secured to the back members . in the preferred embodiment , the bail supports or flanges 34 are spaced a predetermined distance apart indicated by distance a in fig5 . the length of the cylindrical head portions 16 on the ends of legs 14 or bail 12 are predetermined to be slightly shorter than the width a between the wall surfaces 36 as shown in fig1 and 2 . the pins or stud projections 22 project beyond the ends of the heads and beyond the surface of the wall 36 including recess 38 into the recess which has a depth sufficient to receive the pin without binding . the greatest distance across the diagonal of the head portion , denoted by dimension b in fig3 is predetermined to be slightly less than the distance a between the wall surfaces 36 . the greatest distance across the head portion is from one of the corners of the end from which the stud projection 22 extends to the inclined surface 24 forming the cut - away portion on the opposite diagonal corner adjacent the bottom surface of the bail . thus , head portions 16 can be rotated into place while the cut - away portion provided by inclined surface 24 provides clearance room for such rotation as shown in fig3 . in embodiment 30 of the back member , a pair of aligned socket members 42 are integrally cast on the rear surface of base 32 oposite from the surface from which the bail supports or flanges 34 extend . each socket is generally cylindrical but slightly tapered to facilitate removal from its die and includes a bore therein which is threaded to receive a threaded fastener or screw 44 as shown in fig2 . the sockets 42 , which are spaced apart at the rear of the back member , are inserted into bores or pilot holes p through aperture panel f ( fig2 ) and a threaded fastener or screw 44 is inserted from the opposite end of the bore p for receipt in the socket to attach the back member , and thus the back and bail assembly , to the furniture . the spaced location of the socket members 42 prevents rotation or turning of the back member when installed on the furniture . preferably , bail or handle 12 and back members 30 are die cast from zinc or a similar metal . casting allows the various projections to be easily and inexpensively formed without time - consuming and expensive machining and milling operations . in order to facilitate removal from the die , the wall surfaces 36 diverge slightly outwardly from base 32 , as shown in fig3 while the sides of sockets 42 taper slightly toward one another ( fig3 and 6 ). also , recesses 38 extend through and open to the back of base 32 allowing removal of the core used to form the recess when the back member is removed from the die . referring now to fig7 - 10 , another embodiment 50 of the back members for pivotally securing the bail or handle 12 to a furniture component are shown . back members 50 are similar to back members 30 but are die cast from zinc with a shell - like exterior including the various projections and protrusions . back members 50 include a base portion 52 and bail supports or flanges 54 extending outwardly away from the base portion . supports or flanges 54 are integrally connected by a joining flange or wall section 56 extending between like ends of the projections 54 . projections 54 and wall section 56 form a socket - like recess for receipt of the head portion 16 of one of the legs of the bail or handle 12 intermediate generally opposing , planar wall surfaces 58 and immediately adjacent transverse wall section 60 . wall surfaces 58 and 60 are generally perpendicular to base 52 . each of the projections 54 includes an aperture 62 extending outwardly away from base portion 52 having a closed end 64 . these apertures correspond to recess 38 in embodiment 30 of the back member and allow the single back member 50 to be used without being inverted with either leg of the bail or handle 12 regardless of whether the pin or stud projection 22 is on the inside or outside of the leg . such apertures extend through only the walls 58 but not the outer walls of the flanges or bail supports 54 . on the side of base portion 52 , opposite the side from which flanges 54 project , is formed a single socket 66 including a threaded bore for receipt of a threaded fastener 68 inserted through a bore p in a furniture component f as shown in fig9 . in order to prevent rotation or turning of the back member 50 , a tapered , sharp projection 70 is provided at a spaced location from socket 66 for embedding in the surface of the furniture component f . if it is desired to avoid marring or scratching of the surface of the furniture component , or to provide a more decorative base for mounting the back and bail assembly , a separate plate 72 may be mounted behind the back members 50 . such plate may be either flat or curved in cross section to conform to the contour of the furniture component f ( fig8 ). plates 72 includes pairs of spaced , aligned apertures 74 and small detents or apertures 76 for receipt of socket members 66 and projections 70 , respectively , to allow attachment of the back members 50 to the furniture components . for assembly of the bail or handle 12 with a pair of back members of either embodiment 30 or 50 , one of the back members 30 or 50 is attached to the furniture component as shown in fig2 , or 9 . thereafter , the head portion 16 of one leg 14 of bail or handle 12 is inserted between flanges 34 or 54 of the secured back member with the pin or stud projection 22 being inserted in recess 38 or aperture 62 at an angle to the wall surface 36 or 58 . thereafter , the second back member 30 or 50 is likewise assembled to the opposite leg of the bail or handle 12 by inserting pin or stud projection 22 into the recess 38 or aperture 62 in the outside flange 34 of another like back member . the unattached back member is then rotated with respect to its leg such that the head portion 16 is seated between flanges 34 or 54 . the entire bail is then rotated to seat the head portion 16 between the flanges 34 or 54 of the back member which is already secured to the furniture component . such rotation of the entire bail or handle allows the socket members 42 , 66 and projections 70 to be inserted in the furniture component and the fastening screws 44 or 68 threadedly secured therewith to complete the assembly . the cut - away portions defined by inclined surfaces 24 allow such assembly rotation only when bail or handle 12 is subtantially parallel to bases 32 , 52 but not when the bail is perpendicular thereto . thus , the bail will not pull away from the back members when pivoted outwardly from the furniture to open a drawer , cabinet door , or the like on which the assembly is mounted . it will be understood that assembly of both the back members to the individual legs of the bail or handle may also be accomplished before attachment of either back member to the furniture component . alternatively , one of the back members may be first secured to the furniture component or furniture component plus a plate 72 after which the remaining back member and bail are assembled to it . in any event , the cut - away portion formed by inclined surface 24 allows the rotational movement necessary to assemble the back members and bail into the seated position in which the pin or stud projections 22 extend generally transverse to the wall surfaces 36 or 58 and pivotaly engage the closed ends 40 or 64 of recesses 38 or apertures 62 . when seated , head portions 16 are spaced slightly away from bases 32 , 52 of the back members . a small projection 26 may be cast integrally on the rear surface of one or both of legs 14 of bail or handle 12 at a position spaced slightly away from the head portion 16 ( fig4 and 9 ). such projection 26 spaces the bail away from the furniture component to which the bail and back assembly is secured by engaging the base portion 32 or 52 of the back members thereby preventing the bail from marring or scratching the furniture surface . accordingly , the present invention provides a back and bail assembly which gives the impression of each leg of the bail or handle being secured by a pin connection to supports on both sides of each leg . however , the assembly is accomplished with but a single pin or stud projection extending laterally from only one side of each leg thereby greatly simplifying the assembly procedure and allowing such assembly from the front surface of the furniture . the cutaway portions on the surfaces of the leg opposite the pin or stud projections allow rotation of the legs into position between the bail supports or flanges to seat the legs in their assembled positions in a quick and simple manner once the pin or stud projection has been inserted into the recess or aperture provided therefor in the back member . removal of the bail or handle is prevented unless one of the back members is removed from the furniture allowing rotational movement of the bail transverse to the surface of the furniture . while several forms of the invention have been shown and described , other forms will now be apparent to those skilled in the art . therefore , it will be understood that the embodiments shown in the drawings and described above are merely for illustrative purposes , and are not intended to limit the scope of the invention which is defined by the claims which follow .

the specification discloses a furniture pull comprising a back and bail assembly adapted for ease of installation . each of two legs of the bail or handle includes a pin or projection extending laterally to only one side and a cut - away portion formed by a surface inclined to the direction of extension of said pin on the opposite side of the leg . a back member is provided for each leg and includes opposing wall surfaces , one of which includes a recess for the leg pin . for assembly , the pin is inserted in the wall recess and the leg or back member rotated to seat the leg between the walls while the cut - away portion provided by the inclined surface provides clearance room for such rotation . each leg appears to be pinned to both its adjacent wall surfaces while , in fact , it is pivotally secured to only one .

preferably the invention relates to the use of meloxicam or a pharmaceutically acceptable salt thereof for preparing a pharmaceutical composition in a form suitable for systemic or oral administration for the treatment or prevention of respiratory diseases in pigs . meloxicam ( 4 - hydroxy - 2 - methyl - n -( 5 - methyl - 2 - thiazolyl )- 2h - 1 , 2 - benzothiazine - 3 - carboxamide - 1 , 1 - dioxide ) of formula is an active substance which belongs to the group of nsaids ( non - steroidal - anti - inflammatory drugs ). meloxicam and the sodium and meglumine salt thereof ( n - methyl - d - glucamine salt ) are described in ep - a - 0 002 482 ( corresponding to u . s . pat . no . 4 , 233 , 299 ), each of which is hereby incorporated by reference . meloxicam may be used according to the invention in the form of a physiologically acceptable acid addition salt . by physiologically acceptable acid addition salts are meant , according to the invention , the meglumine , sodium , potassium , or ammonium salt , preferably the meloxicam meglumine salt . in a further preferred embodiment , the pharmaceutical composition is administered corresponding to a daily dose of meloxicam ranging from 0 . 01 mg / kg to 5 . 0 mg / kg , preferably from 0 . 1 mg / kg to 3 . 5 mg / kg , in particular from 0 . 2 mg / kg to 2 . 0 mg / kg . the pharmaceutical composition is preferably administered in a form suitable for injection , in particular for intramuscular injection , or in form of water soluble granules for administration via drinking water or as top dressing on feed . a suitable injection formulation is disclosed , for example , in example 25 of ep - a - 0 002 482 . furthermore , such injection solutions may additionally contain excipients selected from among citric acid , lecithin , gluconic acid , tartaric acid , phosphoric acid and edta or the salts thereof as disclosed in the examples 1 to 5 of the international patent application wo 01 / 97813 ( corresponding to u . s . patent app . pub no . 2002 / 0035107 ), each of which is hereby incorporated by reference . moreover , an injection solution of meloxicam for needleless injections is disclosed in the international patent application wo 03 / 049733 ( corresponding to u . s . patent app . pub no . 2003 / 0119825 ), each of which is hereby incorporated by reference . suitable water soluble granules for administration via drinking water or as top dressing on feed are , for example , disclosed in the international patent application pct / ep03 / 11802 ( corresponding to u . s . patent app . pub no . 2004 / 0234596 ), each of which is hereby incorporated by reference . in a preferred embodiment of the invention , the meloxicam granules contain a binder which may be selected from among hydroxypropylmethylcellulose , polyvinylpyrrolidone , gelatine , starch , and polyethylene glycol ether , preferably hydroxypropylmethylcellulose , polyvinylpyrrolidone , and polyethylene glycol ether , and most preferably hydroxypropylmethylcellulose and polyvinylpyrrolidone . in another preferred embodiment of the invention , meloxicam granules contain a sweetener , which may be selected from among sodium saccharine , aspartame , and sunett ® ( acesulfame k ), preferably sodium saccharine or aspartame . particularly preferred according to the invention are meloxicam granules containing a flavoring agent which may be selected from among vanilla , honey flavoring , apple flavoring , and contramarum , preferably honey flavoring and apple flavoring . also particularly preferred are meloxicam granules in which the carrier is selected from among lactose , glucose , mannitol , xylitol , sucrose , and sorbitol , preferably glucose , lactose , or sorbitol , more preferably glucose or lactose , and most preferably glucose . particularly preferred are meloxicam granules in which the content of meloxicam is between 0 . 05 % and 4 %, preferably between 0 . 1 % and 2 %, preferably between 0 . 3 % and 1 . 8 %, more preferably between 0 . 4 % and 1 . 5 %, and most preferably 1 . 2 %. also particularly preferred are meloxicam granules which contain meglumine and meloxicam in a molar ratio of about 9 : 8 to 12 : 8 , preferably 10 : 8 . meloxicam can be used according to the invention to treat or prevent respiratory diseases in any breed of swines . preferably pigs selected from the swine breeds american landrace , american yorkshire , angeln saddleback , arapawa island , ba xuyen , bantu , bazna , beijing black , belarus black pied , belgian landrace , bentheim black pied , berkshire , black slavonian , british landrace , british lop , bulgarian white , cantonese , chester white , czech improved white , danish landrace , dermantsi pied , duroc , dutch landrace , fengjing , finnish landrace , french landrace , german landrace , gloucestershire old spots , guinea hog , hampshire , hereford , hezuo , iberian , italian landrace , jinhua , kele , krskopolje , kunekune , lacombe , large black , large black - white , large white , lithuanian native , mangalitsa , meishan , middle white , minzhu , mong cai , mukota , mora romagnola , moura , mulefoot , neijiang , ningxiang , norwegian landrace , ossabaw island , oxford sandy and black , philippine native , pietrain , poland china , red wattle , saddleback , spots , swabian - hall , swedish landrace , tamworth , thuoc nhieu , tibetan , turopolje , vietnamese potbelly , welsh , and wuzhishan , in particular american landrace , belgian landrace , british landrace , danish landrace , dutch landrace finnish landrace , french landrace , german landrace , italian landrace , and pietrain can be treated with meloxicam according to the present invention . furthermore preferred is the administration of meloxicam is in conjunction with an antibiotic , preferably selected from the group consisting of β - lactams , quinolones , tetracyclines , sulfonamides , fenicoles , and macrolides . most preferred are amoxicillin , oxytetracycline , florfenicol , tylosin , tilmicosin , and sulfamethazine . the dose of antibiotic is not critical per se and depends strongly on the different efficacies of the antibiotics used . as a rule up to 150 . 0 mg / kg , preferably from 0 . 1 mg / kg to 120 mg / kg , in particular from 10 mg / kg to 110 mg / kg of an antibiotic are co - administered together with meloxicam . amoxicillin : 5 mg / kg to 30 mg / kg , in particular about 10 mg / kg ; oxytetracycline : 20 mg / kg to 70 mg / kg , in particular about 30 mg / kg ; florfenicol : 10 mg / kg to 20 mg / kg , in particular about 15 mg / kg ; tylosin : 10 mg / kg to 25 mg / kg , in particular about 16 mg / kg ; tilmicosin : 5 mg / kg to 30 mg / kg , in particular 10 mg / kg to 20 mg / kg ; and sulfamethazine : 80 mg / kg to 150 mg / kg , in particular about 100 mg / kg . the phrase “ co - administration ” ( or “ administration in conjunction with ”), in defining use of meloxicam and an antibiotic , is intended to embrace administration of each agent in a sequential manner in a regimen that will provide beneficial effects , in particular , reduction of the symptoms of the respiratory disease in the affected pig of the drug combination . the phrase also is intended to embrace co - administration of these agents in a substantially simultaneous manner , such as in a single capsule or injection solution having a fixed ratio of these active agents or in multiple , separate capsules for each agent . accordingly , meloxicam and the antibiotic may be co - administered in a combined form , or separately or separately and sequentially wherein the sequential administration is preferably close in time . preferably the medicament according to this invention is used for the prevention or treatment of porcine respiratory disease complex in growing or fattening pigs ; or for the prevention or treatment of respiratory diseases in pigs caused by mycoplasmas , in particular mycoplasma hyopneumoniae , mycoplasma hyorhinis , for the prevention or treatment of respiratory diseases in pigs caused by bacteria in particular actinobacillus spp ., in particular actinobacillus pleuropneumoniae , bordetella bronchiseptica , pasteurella multocida , arcanobacterium pyogenes , streptococcus spp ., and staphylococcus spp ., or for the prevention or treatment of respiratory diseases in pigs caused by viruses , in particular swine influenza virus , aujetzky &# 39 ; s virus , porcine reproductive and respiratory syndrome virus , porcine circovirus , and transmissible gastroenteritis and porcine respiratory coronavirus . most preferably the medicament according to this invention is used for the prevention or treatment of respiratory diseases in pigs caused by mycoplasma hyopneumoniae , actinobacillus pleuropneumoniae , bordetella bronchiseptica , pasteurella multocida , streptococcus suis , swine influenza virus , and porcine reproductive and respiratory syndrome virus . the examples that follow serve to illustrate the use of meloxicam according to the invention . they are intended solely as possible procedures described by way of example , without restricting the invention to their content . efficacy of meloxicam in pigs with experimental actinobacillus pleuropneumoniae infection the study was a controlled , randomized , and blinded exploratory study under experimental conditions with a parallel group design . crossbred pigs of about 10 weeks of age were challenged with a single intranasal inoculation of actinobacillus pleuropneumoniae . the next day , pigs were included in the study and treated if they fulfilled the following inclusion criteria : rectal temperature ≧ 40 ° c . and clinical symptoms of acute or subacute infectious respiratory disease . twenty - four ( 12 castrated male and 12 female ) pigs were included and randomly allocated to three treatment groups with 8 pigs per group . the treatment groups were : meloxicam was administered as 0 . 5 % solution , at 0 . 5 mg / kg daily on three consecutive days , oxytetracycline as 20 % long - acting solution ( oxytet ® 200 ) at 20 mg / kg as single injection . relevant criteria for the evaluation of efficacy were incidence of fever , clinical parameters of respiratory disease , deaths , and lung lesions at necropsy 10 days after first treatment or after spontaneous death . the percentage of affected lung tissue was calculated by lobe and averaged for the total lung . challenge with actinobacillus pleuropneumoniae lead to severe pleuropneumonia within 12 hours . the incidence of fever ( rectal temperature ≧ 40 . 56 ° c .) following the first treatment was lower in group 3 (♦) than in groups 1 ( δ ), and 2 (∘) ( cp . fig1 ). the best treatment response in clinical parameters was observed in group 3 . the number of pigs which died during the three days following first treatment is displayed below . group ( n = 8 per group ) deaths 1 7 2 1 3 0 the mean extent of lung lesions was less severe in group 3 than in the other groups ( see below ). group lung lesions (%) 1 60 2 35 3 14 meloxicam in addition to antibiotic treatment effectively reduced fever , clinical symptoms of respiratory disease , deaths , and the extent of lung lesions in pigs with experimental actinobacillus pleuropneumoniae - infection . efficacy of meloxicam in drinking water in experimental swine influenza virus infection the aim of this study was to test the efficacy of meloxicam granules dissolved in drinking water in pigs experimentally infected with swine influenza virus ( siv ). the study was an open , negative controlled randomized laboratory study carried out according to gcp at one site . meloxicam granules containing 6 mg meloxicam per gram were offered to the pigs in the treatment groups ( a + b ) via drinking water in a concentration of 1 g granules per liter drinking water ad libitum for 7 consecutive days . this resulted in an actual meloxicam uptake of 0 . 8 mg per kg body weight per day . the pigs in the control group ( c ) received municipal drinking water ad libitum . 30 pigs were infected with siv on study day 0 . 10 pigs were allocated to each of the three groups a , b , and c . treatment ( groups a and b ) started after siv challenge on the same day . the study animals were clinically examined daily on study days 0 to 7 and 14 . they were weighed on study days 7 and 14 . all animals of group a and 5 animals of group c were euthanized and necropsied on study day 7 ; the remaining study animals , group b and 5 study animals of group c , on study day 14 . it is the major finding of this study that meloxicam granules administered continuously in the drinking water at an approximate daily dose of 0 . 8 mg / kg body weight significantly alleviated the development of lung lesions caused by experimental infection with siv during the first week after challenge . fig2 shows the quantity of lung lesions by lung lobe on study days 7 and 14 . on study day 7 the percentage of lung tissue affected with siv - related lesions ( median value ) was 8 . 9 % in meloxicam group a and 23 . 8 % in the control group ( 5 study animals of group c ). moreover , meloxicam - treated pigs reached significantly higher weight gains during the two weeks following infection than untreated controls . mean daily weight gain in the interval study day 0 to 7 was 557 g in meloxicam group a and 257 g in the control ( 5 study animals of group c ). in the interval study day 0 to 14 , mean daily weight gain was 629 g in meloxicam group b and 486 g in the control ( 5 study animals of group c ). the area under the curve of the clinical index score ( cis ), a sum of the relevant clinical parameters , over study days 0 to 7 was significantly smaller in groups a and b than in group c . thus oral treatment with meloxicam granules at a dose of 0 . 8 mg meloxicam per kg body weight per day for 7 consecutive was an efficacious treatment for siv infection . field trial regarding the effect of meloxicam in the porcine respiratory disease complex ( prdc ) in growing / fattening pigs a medium scale farm ( 560 sows ) with a previous history of recurring prdc episodes was selected . a double - blinded randomized study was carried out with the selection of 162 growing animals with a mean age of 90 days at the onset of prdc clinical signs . animals were randomly allocated to 8 pens and divided into two treatment groups , with respect to equal sex ratio , same housing and feeding conditions and genetic background . group 1 ( pc ) received 800 ppm chlorotetracycline in the feed over 8 consecutive days plus a single im injection of a placebo ( isotonic saline ) at d 0 ( start of the trial , n = 82 ). group 2 ( m ) received 800 ppm chlorotetracycline in the feed over 8 consecutive days plus a single im injection of 0 . 4 mg / kg bodyweight meloxicam ( metacam ® 2 %, boehringer ingelheim gmbh ) at d 0 ( n = 80 ). clinical parameters were assessed as the daily respiratory score ( rs ), using a 3 point score ( 0 = absence of signs to 3 = abdominal breathing and disordered general condition ) over 8 consecutive days and the total number of additional required injectable medications ( aim ). growth performance data for each group included the average daily gain ( adg ) for the following trial periods : d90 to d117 , d117 to d170 ( slaughtering ), and d90 to d170 of age . mortality was also calculated for these time periods . slaughterhouse records per group , included the percentage of each lung surface ( ls ) affected by chronic and acute respiratory lesions . student &# 39 ; s t - test and pearson &# 39 ; s chi - square test were used for the consequent comparisons of means and frequencies between trial groups . rs and aim in the meloxicam group were significantly lower ( p & lt ; 0 . 05 ) compared to the control group . same applies for ls affected by acute lesions ( p & lt ; 0 . 01 ), while no differences were observed for ls in chronic cases ( table 1 ). table 2 adg : mean ( sd ) trial period group d90 to d117 d117 to d170 d90 to d170 pc 0 . 64 ( 0 . 09 ) a 0 . 89 ( 0 . 06 ) a 0 . 81 ( 0 . 03 ) a m 0 . 67 ( 0 . 10 ) b 0 . 89 ( 0 . 06 ) a 0 . 82 ( 0 . 03 ) a a , b values in a column with different superscripts differ significantly ( p & lt ; 0 . 05 ) under the conditions of this study , the reduction of the prevalence of respiratory signs as well as the reduced overall number of required injectable antibiotic medications are indicative for the potent anti - inflammatory activity of meloxicam . the latter could become a valuable adjunctive measure , especially when respiratory distress is associated with remarkable reduction of the feed intake . the initial differences in growth performance and in mortality rate could be explained by the fact that meloxicam , when combined with proper antimicrobial medication , contributes to faster recovery from a respiratory inflammation and faster restoring of the distorted growth rate of affected animals . further research on the evaluation of feed intake and the use of meloxicam in prdc recurring episodes is required .

a method of treating or preventing a respiratory disease in a pig , the method comprising administering to the pig in need thereof an effective amount of meloxicam or a pharmaceutically acceptable salt thereof .

the objects , features , and advantages of the inventive concept presented in this application are more readily understood when referring to the accompanying drawings . the drawings , totaling five figures , show the basic components and functions of the device and methods of use . in the several figures , like reference numbers are used in each figure to correspond to the same component as may be depicted in other figures . the discussion of the present inventive concept will be initiated with fig1 , which illustrates a front view of the seating device 1 in its open configuration . in the open configuration , the seating device 1 is receptive to a user 40 ( not shown ) sitting upon the seating device 1 . the essential components of the seating device 1 , as visible in fig1 , comprise a top panel 13 , upon which a user &# 39 ; s 40 thighs 45 , 46 will rest ; a contoured recess 12 , which is contoured to correspond generally to a user &# 39 ; s 40 buttocks 41 , 42 ; an inner backrest 10 , a back panel 11 ; a horizontally - oriented space 17 ; a left hinge 6 [ shown in fig5 ( c ) ] and left hinge bolt 8 ; a right hinge 7 ( a right hinge bolt 9 is out of view ); left and right armrests 2 , 3 ; and left and right armrest supports 4 , 5 . the contoured recess 12 is a solid , one - piece structure further comprising a top panel 13 , a left side panel 15 , a right side panel 16 not shown in fig1 ), a forward front panel 14 , a rear panel rear panel 18 , and a continuous bottom edge 21 . fig2 presents a left - side view of the seating device 1 , again in the open configuration . the rightmost portion of fig2 shows the inner backrest 10 and the back panel 11 . the inner backrest 10 comprises an elastomeric exterior surface which encloses an underlayment of cushioning material , while the back panel 11 is composed of a rigid material for vertical structural integrity . the inner backrest 10 and back panel 11 are permanently joined in an abutting manner at a common interfacing surface 20 . fig2 also shows the lower section of the seating device 1 , which essentially is the contoured recess 12 . as shown in fig2 , the contoured recess 12 comprises a solid , box - like structure having a front panel 14 , a top panel 13 , a left side panel 15 , ( the right side panel 16 is out of view ), a rear panel 18 , and a bottom edge 21 , which edge 21 is common to the bottom of the left and right side panels 15 , 16 , the bottom of the front panel 14 , and the bottom of the rear panel 18 . the left armrest 2 is shown , being permanently attached at the angle shown , to a left armrest support 4 . in turn , the left armrest support 4 is permanently attached to the left side of the top panel 13 . in a corresponding manner , the right armrest 3 ( not shown ) is attached to a right armrest support 5 , the right armrest support 5 being attached to the right side of the top panel 13 . in fig2 , the contoured recess 12 is shown to be an arcuate , transverse padded channel which runs the entire width from the left side to the right side of the seating device 1 . the contoured recess 12 is a continuous structure shaped so as to generally correspond to the roundness of a user &# 39 ; s buttocks . again referring to fig2 , the upper half of the left hinge plate 6 is permanently attached to the left side of , and parallel to , the back panel 11 . both hinge plates 6 , 7 , may also be manufactured as an integral part of the back panel 11 . the left hinge plate 6 is constructed with an aperture proximate the lower end of the left hinge 6 so as to accommodate the insertion of a left hinge bolt 8 . by the same design , the upper half of the right hinge 7 [ shown in fig5 ( c ) ] is permanently attached to the right side of , and parallel to , the back panel 11 . the right hinge plate 7 is constructed with an aperture proximate the lower end of the right hinge plate 7 so as to accommodate the insertion of a right hinge bolt 9 . in fig2 , both sides of the contoured recess 12 , near the rear panel 18 , are constructed with two threaded horizontal openings : one on the upper left side panel 15 and one on the right side panel 16 . the threaded horizontal openings correspond to the threading of the previously - mentioned left hinge bolt 8 and the right hinge bolt 9 , respectively . during manufacture , the back panel 11 , having the left hinge plate 6 and right hinge plate 7 permanently attached , is lifted into a position directly above the rear panel 18 , so as to align the respective apertures of the left hinge plate 6 and right hinge plate 7 with the threaded openings on the left - side and right - side panels 15 , 16 . the left hinge bolt 8 and right hinge bolt 9 are then inserted through the respective hinge apertures and into the threaded openings on the left - side and right - side panels 15 , 16 . the left and right hinge plates 6 , 7 , may be of a variety of different designs and construction , including an arrangement providing a spring - loaded attachment to the back panel 18 or to both side panels 15 , 16 of the contoured recess 12 . further , the left and right hinge plates in an alternative embodiment , the contoured recess 12 is configured with a horizontally - oriented circular channel extending the width of the seating device 1 from the rearmost portion of the left side panel 15 to the rearmost section of the right side panel 16 . a corresponding circular , cross - sectioned rod 22 having a diameter corresponding to the inner diameter of said width - wise circular channel and a length corresponding to the length of said horizontally - oriented circular channel is included in the alternative embodiment . the said circular rod 22 further features exterior threading at both ends . the circular rod 22 , once inserted into the circular channel , utilizes two threaded flat - head nuts 23 , shown in fig5 ( a ) , having internal threading corresponding to the exterior threads of said circular rod 22 , to fasten both ends of the circular rod 22 securely against the left side panel 15 and right side panel 16 of the contoured recess 12 . viewing fig3 , there is shown a user 40 in the motion of lowering the body onto the seating device 1 , with the left hamstring muscles 45 approaching contact with the top panel 13 of the contoured recess 12 . the left arm 43 of the user 40 assists in giving balance during the seating process by providing a gradual shift of the user &# 39 ; s 40 center of gravity . once seated , the user &# 39 ; s back 47 will make contact with the cushioned inner back rest 10 , and the buttocks 41 will be suspended directly over the concave portion of the contoured recess 12 . fig4 illustrates a situation where the seating device 1 has been placed atop a chair 30 commonly found in residential or commercial settings . the back panel 11 of the seating device 1 is placed firmly against the chair back 33 and the bottom of the contoured recess 12 is placed in a level position upon the chair 30 , corresponding to the horizontal of support provided by the chair 30 legs 34 . a fully - seated user 40 is shown from the perspective of the right side of the seating device 1 . the right hamstring muscles 46 of the user 40 are shown to be firmly supported by the top panel 13 of the contoured recess 12 , and the back 47 of the user 40 may be firmly pressed against the cushioned back rest 10 . the user &# 39 ; s 40 buttocks 42 are seen to be suspended directly above the concave segment of the contoured recess 12 . fig5 depicts the seating device 1 in its compacted configuration , in which the back panel 11 and cushioned inner backrest 10 have been folded down from their previous vertical orientation into a horizontal orientation abutting the top panel 13 of the contoured recess 12 . while preferred embodiments of the present inventive concept have been shown and disclosed herein , it will be obvious to those persons skilled in the art that such embodiments are presented by way of example only , and not as a limitation to the scope of the inventive concept . numerous variations , changes , and substitutions may occur or be suggested to those skilled in the art without departing from the intent , scope , and totality of this inventive concept . such variations , changes , and substitutions may involve other features which are already known per se and which may be used instead of , in combination with , or in addition to features already disclosed herein . accordingly , it is intended that this inventive concept be inclusive of such variations , changes , and substitutions , and by no means limited by the scope of the claims presented herein .

the disclosed invention provides a seating device for use by individuals who have recently had a surgical procedure that makes it uncomfortable or painful for the person to bear their body weight on their buttocks region . the device includes a firm back support and a cushioned bottom portion having a contoured recess thereon , thereby allowing an individual to place their body weight on the thigh area rather than directly onto the buttocks , offering comfort while sitting . the back support is connected to the bottom portion by a hinge , enabling the back support to fold down on the seat for easy and convenient portability . overall , the present invention allows people to go about their daily routines without worry of additional discomfort when sitting down at home , at work , during vehicular transit , or other situations .

the brush is designated in its entirety by the letter b and comprises , as its basic elements , a handle h and a brush insert i . both the handle and insert are fabricated of a relatively high density polymer , such as 0 . 0903 density polypropylene . the handle is of an integral construction and formed with an elongate gripping portion 10 having an open loop 12 of an ovoid configuration formed at one end thereof . the gripping portion is of a slightly arcuate configuration , as may be seen from fig1 and has a finger depression 14 formed on one side . the loop 12 is symmetrical and formed with an internal shoulder 16 at one side thereof . to the side of the shoulder the loop is formed with a slightly divergent surface 18 . ideally the divergence is approximately 120 , as seen by the angle α in fig1 . the insert i is of an ovoid configuration complemental to that of the interior of the loop 12 and comprises : a body section 20 having a slightly tapered exterior surface adapted to mate with the interior surface 18 of the loop ; a 360 ° flange 22 adapated to sealingly mate with one side of the loop 12 when the brush insert is fully in place within the loop ; and , a 360 ° bead 24 adapated to snap past and engage the shoulder 16 to the side of the handle opposite that engaged by the flange 22 . in the preferred embodiment , the side of the insert on which the bead 24 is formed extends approximately 0 . 030 of an inch beyond the side of the handle when the insert is fully engaged within the loop ( see fig2 c ). such projection of the insert from the handle assures that the insert is securely held in place within the loop and also provides a raised surface on the insert to which pressure may be applied to eject the insert from the loop . fig2 a shows the insert and handle in an exploded - view aligned with one another . fig2 b shows the insert as it is moved into the loop from the exploded condition shown in fig2 a . fig2 c shows the insert after it has been pushed fully into the loop , with the bead 24 snapped past and into secured sealed engagement with the shoulder 16 . in the latter condition , the flange 22 is sealingly engaged with the side of the loop opposite of that engaged by the bead . for purposes of description , the side of the loop engaged by the bead is designated by the numeral 26 and that engaged by the flange 22 is designated by the numeral 28 . bristles 30 are shown extending from the side of the insert about which the flange 22 is formed . in the preferred embodiment , a slightly beveled surface 32 is formed on the portion of the shoulder 16 which opens through the side 26 . this surface facilitates snapping of the insert into and out of the loop and also provides a smooth surface against which the bead 24 may seat to form a seal . fig4 illustrates the manner in which the bead 24 engages the beveled surface 32 of the shoulder 16 through a full 360 °. fig5 similarly shows the manner in which the flange 22 engages the side 28 through a full 360 °. from these figures , and the cross - sectional views of fig2 c and fig3 it can be seen that the engaged insert is fully sealed to the loop of the handle about a full 360 ° on both sides of the handle . the process of ejecting the insert from the handle is essentially a reverse of the insertion procedure . in this process , with the insert fully engaged as shown in fig2 c , pressure is applied to move the insert outwardly of the loop as shown in fig2 b . continued pressure functions to fully eject the insert , as shown in fig2 a . to facilitate ejection , the user would typically press on one of the narrower ends of the insert to get the injection step started . in this way , the pressure required to initiate snapping of the insert out of the loop is minimized . once one end of the bead is disengaged from the shoulder , the remainder of the insert easily follows . the symmetrical ovoid configurations of the mating insert and loop enable the insert to be placed in either of two positions rotated 180 ° from one another . this simplifies the insertion process in that the user simply has to align the insert with the loop and does not need to be concerned about which end of the insert is at which end of the loop . it also enables the insert to be turned 180 ° after the brush has been in use . such turning enables the user to rotate the brush if the bristles on one side are wearing faster than those on the other . the present invention provides an improved polymeric toothbrush with a removable insert . it is particularly advantagous in that when in place the insert is fully sealed within the handle and yet may be readily removed therefrom . it should be appreciated , however , that the invention is not intended to be limited to the specifics of the illustrated embodiment , but rather as defined by the accompanying claims .

a tooth brush having a replaceable brush insert complementally receivable in an open loop formed at one end of an elongate handle . the loop is of a symmetrical ovoid configuration . a bead extends fully around the insert and snaps to one side of the loop to secure the insert within the loop . a flange extends around the insert for sealed engagement with the loop upon secure receipt of the insert within the loop .

in the embodiment of the invention discussed below , the tachycardia and fibrillation detection criteria discussed above in conjunction with the medtronic model 7216 and model 7217 implantable pacemaker / cardioverter / defibrillators are employed , and the following discussion of the present invention should be understood in this context , with the present invention being employed in this embodiment to adjust the dividing point fdi between the interval ranges associated with ventricular tachycardia and ventricular fibrillation . however , while the specific embodiment disclosed below is directed to distinguishing between ventricular tachycardia and fibrillation , it is also believed that the invention may also be usefully be practiced in the context of a device for treating atrial tachyarrhythmias . moreover , the value of the present invention is not limited to the context of the specific detection criteria disclosed , but is believed workable and valuable in the context of any devices which distinguish between tachycardia and fibrillation using rate or interval based criteria . fig1 illustrates the relationship between the interval ranges associated with detection of fibrillation and tachycardia , as employed in the context of the present invention . the maximum interval indicative of fibrillation ( minimum interval indicative of tachycardia ), &# 34 ; fdi p &# 34 ; is defined during programming of the device . the maximum interval indicative of tachycardia , tdi , is similarly defined during programming of the device . as illustrated , an increments to ( fdi p plus delta ) or decrements from ( fdi p minus delta ) the value the value of fdi p may occur as part of the detection process including the present invention . initially , fdi p serves as the current maximum interval indicative of fibrillation ( fdi c ). as increments or decrements are made , the incremented or decremented values are used as fdi c . increments are only allowable up to the point where the incremented value would exceed a maximum value ( fdi max ). decrements are only allowable up to the point where the decremented value would be less than a minimum value ( fdi min ). in the context of the preferred embodiment of the present invention , four or eight preceding r -- r intervals less than tdi , for example , may be examined to determine whether the duration of fdi c needs to be incremented or decremented . the value of the increment delta may be , for example , 10 to 30 ms , and the values of fdi max and fdi min may be , for example , fdi c plus 20 to 60 milliseconds and fdi c minus 20 to 60 milliseconds , respectively . alternatively , the invention may be practiced in a fashion such that fdi p defines either the maximum value or the minimum value of fdi c . for example , the physician may wish to allow the device only to become more biased toward detection of ventricular fibrillation , as compared to detection using the programmed rate interval ranges . in this case , fdi min would be set equal to fdi p . fdi c may be incremented , for example , in response to more than fifty percent or more of the n intervals being less than fdi c and fdi c may correspondingly be decremented in response to more than fifty percent of the n intervals being greater than or equal to fdi c . alternatively , more stringent criteria for incrementing and decrementing fdi c may be applied , with incrementing occurring only in response to seventy - five percent or more of the n intervals being less than fdi c and decrementing occurring only in response to seventy - five percent or more of the n intervals being greater than or equal to fdi c . it is anticipated that in greater than or equal to fdi c . it is anticipated that in commercial embodiments of the present invention , some or all of the values and parameters discussed above will be selectable by the physician . fig2 is a functional schematic diagram of an implantable pacemaker / cardioverter / defibrillator in which the present invention may usefully be practiced . this diagram should be taken as exemplary of the type of device in which the invention may be embodied , and not as limiting , as it is believed that the invention may usefully be practiced in a wide variety of device implementations , including devices having functional organization similar to any of the implantable pacemaker / defibrillator / cardioverters presently being implanted for clinical evaluation in the united states . the invention is also believed practicable in conjunction with implantable pacemaker / cardioverters / defibrillators as disclosed in prior u . s . pat . no . 4 , 548 , 209 , issued to wielders , et al . on oct . 22 , 1985 , u . s . pat . no . 4 , 693 , 253 , issued to adams et al . on sep . 15 , 1987 , u . s . pat . no . 4 , 830 , 006 , issued to haluska et al . on may 6 , 1989 and u . s . pat . no . 4 , 949 , 719 , issued to pless et al . on aug . 21 , 1990 , all of which are incorporated herein by reference in their entireties . the device is illustrated as being provided with six electrodes , 500 , 502 , 504 , 506 , 508 and 510 . electrodes 500 and 502 may be a pair of endocardial electrodes located in the ventricle , mounted to a transvenous lead . electrode 504 may correspond to a remote , indifferent electrode located on the housing of the implantable pacemaker / cardioverter / defibrillator . electrodes 506 , 508 and 510 may correspond to the large surface area defibrillation electrodes located on ventricular , coronary sinus , superior vena cava or subcutaneous leads or to epicardial defibrillation electrodes . electrodes 500 and 502 are shown as hard - wired to the 514 , auto threshold circuit 516 for providing an adjustable sensing threshold as a function of the measured r - wave amplitude and comparator 518 . a signal is generated on r - out line 564 whenever the signal sensed between electrodes 500 and 502 exceeds the present sensing threshold defined by auto threshold circuit 516 . as illustrated , the gain on the band pass amplifier 514 is also adjustable by means of a signal from the pacer timing and control circuitry 520 on gain adj line 566 . the operation of this r - wave detection circuitry may correspond to that disclosed in commonly assigned , co - pending u . s . patent application ser . no . 07 / 612 , 760 , by keimel , et al ., filed november 15 , for an apparatus for monitoring electrical physiologic signals , incorporated herein by reference in its entirety . however , alternative r - wave detection circuitry such as that illustrated in u . s . pat . no . 4 , 819 , 643 , issued to menken on apr . 11 , 1989 and u . s . pat . no . 4 , 880 , 004 , issued to baker et al . on nov . 14 , 1989 , both incorporated herein by reference in their entireties , may also usefully be employed to practice the present invention . the threshold adjustment circuit 516 sets a threshold corresponding to a predetermined percentage of the amplitude of a sensed r - wave , which threshold decays to a minimum threshold level over a period of less than three seconds thereafter , similar to the automatic sensing threshold circuitry illustrated in the article &# 34 ; reliable r - wave detection from ambulatory subjects &# 34 ;, by thakor et al ., published in biomedical science instrumentation , vol . 4 , pp 67 - 72 , 1978 , incorporated herein by reference in its entirety . it is preferable that the threshold level not be adjusted in response to paced r - waves , but instead should continue to approach the minimum threshold level following paced r - waves to enhance sensing of low level spontaneous r - paced r - waves to enhance sensing of low level spontaneous r - waves associated with tachyarrhythmias . the time constant of the threshold circuit is also preferably sufficiently short so that minimum sensing threshold may be reached within 1 - 3 seconds following adjustment of the sensing threshold equal to 70 - 80 % of the amplitude of a detected spontaneous r - wave . the invention may also be practiced in conjunction with more traditional r - wave sensors of the type comprising a band pass amplifier and a comparator circuit to determine when the band - passed signal exceeds a predetermined , fixed sensing threshold . switch matrix 512 is used to select which of the available electrodes are coupled to band pass amplifier 534 . selection of which two electrodes are so coupled is controlled by the microprocessor 524 via data / address bus 540 . signals from the selected electrodes are passed through band - pass amplifier 534 and into multiplexer 532 , where they are converted to multi - bit digital signals by a / d converter 530 , for storage in random access memory 526 under control of direct memory address circuit 528 . microprocessor 524 analyzes the digitized egm signal stored in random access memory 526 to determine the width of the stored r - wave or in conjunction with the tachycardia / fibrillation discrimination function discussed below . amplifier 534 may be a broad band pass amplifier , having a band pass extending for approximately 0 . 5 to 200 hertz . the filtered egm signal from amplifier 534 is passed through multiplexer 532 , and digitized in a - d converter circuitry 530 . the digitized egm data is stored in random access memory 526 under control of direct memory address circuitry 528 . preferably , a portion of random access memory 526 is configured as a looping or buffer memory which stores at least the preceding several seconds of the egm signal . the data stored in the buffer memory may be optionally employed to perform r - wave width measurements as disclosed in co - pending u . s . patent application ser . no . 07 / 867 , 931 , filed apr . 13 , 1992 by mader et al ., incorporated herein by reference in its entirety and / or to perform the ventricular fibrillation / ventricular tachycardia discrimination function disclosed in pending u . s . patent application ser . no . 07 / 750 , 679 filed aug . 27 , 1991 by bardy et al ., also incorporated herein by reference in its entirety . however , the present invention is readily practiced in devices which do not include such functions , and for purposes of the disclosed preferred embodiment of the present invention it should be assumed that such functions , if available , are programmed off . the microprocessor also updates counts related to the r -- r intervals previously sensed . the counts are incremented on the occurrence of a measured r -- r intervals falling within associated rate ranges . as discussed above these ranges may include the ranges illustrated above in fig1 associated with ventricular tachycardia and ventricular fibrillation , and the stored counts may include vtec and vfec . these rate ranges may be defined by the programming stored in the ram 526 . these counts , along with other stored information reflective of the previous series of r -- r intervals such as information regarding the rapidity of onset of the detected short r -- r intervals , the stability of the detected r -- r intervals , the duration of continued detection of short r -- r intervals , the average r -- r interval duration and information derived from analysis of stored egm segments are used to determine whether tachyarrhythmias are present and to distinguish between different types of tachyarrhythmias , as discussed above in conjunction with fig1 . other such detection algorithms for recognizing tachycardias are described in the above cited u . s . pat . no . 4 , 726 , 380 , issued to vollmann , u . s . pat . no . 4 , 880 , 005 , issued to pless et al . and u . s . pat . no . 4 , 830 , 006 , issued to haluska et al ., incorporated by reference in their entireties herein . an additional set of tachycardia recognition methodologies is disclosed in the article &# 34 ; on - set and stability for ventricular tachyarrhythmia detection in an implantable pacer - cardioverter - defibrillator &# 34 ; by olson et al ., published in computers in cardiology , oct . 7 - 10 , 1986 , ieee computer society press , pp . 167 - 170 , also incorporated by reference in its entirety herein . however , other criteria may also be measured and employed in conjunction with the present invention . it is envisioned that onset and stability requirements are optional in a device employing the present invention , and preferably are made available as programmable options , which may be deleted by external programmer command . if included , it is believed preferable that the onset criteria be required to met prior to initiating counting of vtec , and that once met , the criteria will remain satisfied until detection of tachycardia termination . thus , onset is not intended to be a detection criteria required for re - detection of tachycardia , following initial detection . the width criterion , if used , should also be understood to be used both in initial detection of tachycardia and in re - detection of tachycardia . this reflects a presumption that following initial detection of ventricular tachycardia , absent a proven return to normal heart rhythm ( termination detect ), subsequent high ventricular rates should be presumed to be ventricular in origin . the stability criterion , on the other hand , is believed to be appropriate for use both in initial detection of tachycardia and in re - detection of tachycardia . the remainder of the circuitry is dedicated to the provision of cardiac pacing , cardioversion and defibrillation therapies . the pacer timing / control circuitry 520 includes programmable digital counters which control the basic time intervals associated with vvi mode cardiac pacing , including the pacing escape intervals , the refractory periods during which sensed r - waves are ineffective to restart timing of the escape intervals and the pulse width of the pacing pulses . the durations of these intervals are determined by microprocessor 524 , and are communicated to the pacing circuitry 520 via address / data bus 540 . pacer timing / control circuitry also determines the amplitude of the cardiac pacing pulses and the gain of band - pass amplifier , under control of microprocessor 524 . during vvi mode pacing , the escape interval counter within pacer timing / control circuitry 520 is reset upon sensing of an r - wave as indicated by a signal on line 564 , and on timeout triggers generation of a pacing pulse by pacer output circuitry 522 , which is coupled to electrodes 500 and 502 . the escape interval counter is also reset on generation of a pacing pulse , and thereby controls the basic timing of cardiac pacing functions , including anti - tachycardia pacing . the duration of the interval defined by the escape interval timer is determined by microprocessor 524 , via data / address bus 540 . the value of the count present in the escape interval counter when reset by sensed r - waves may be used to measure the duration of r -- r intervals , to detect the presence of tachycardia and to determine whether the minimum rate criteria are met for activation of the width measurement function . microprocessor 524 operates as an interrupt driven device , and responds to interrupts from pacer timing / control circuitry 520 corresponding to the occurrence of sensed r - waves and corresponding to the generation of cardiac pacing pulses . these interrupts are provided via data / address bus 540 . any necessary mathematical calculations to be performed by microprocessor 524 and any updating of the values or intervals controlled by pacer timing / control circuitry 520 take place following such interrupts . in the event that a tachyarrhythmia is detected , and an anti - tachyarrhythmia pacing regimen is desired , appropriate timing intervals for controlling generation of anti - tachycardia pacing therapies are loaded from microprocessor 524 into the pacer timing and control circuitry 520 , to control the operation of the escape interval counter and to define refractory periods during which detection of an r - wave by the r - wave detection circuitry is ineffective to restart the escape interval counter . similarly , in the event that generation of a cardioversion or defibrillation pulse is required , microprocessor 524 employs the counters to in timing and control circuitry 520 to control timing of such cardioversion and defibrillation pulses , as well as timing of associated refractory periods during which sensed r - waves are ineffective to reset the timing circuitry . in response to the detection of fibrillation or a tachycardia requiring a cardioversion pulse , microprocessor 524 activates cardioversion / defibrillation control circuitry 554 , which initiates charging of the high voltage capacitors 556 , 558 , 560 and 562 via charging circuit 550 , under control of high voltage charging line 552 . the voltage on the high voltage capacitors is monitored via vcap line 538 , which is passed through multiplexer 532 , and , in response to reaching a predetermined value set by microprocessor 524 , results in generation of a logic signal on cap full line 542 , terminating charging . thereafter , delivery of the timing of the defibrillation or cardioversion pulse is controlled by pacer timing / control circuitry 520 . one embodiment of an appropriate system for delivery and synchronization of cardioversion and defibrillation pulses , and controlling the timing functions related to them is disclosed in more detail in co - pending , commonly assigned u . s . patent application ser . no . 07 / 612 , 761 , by keimel , for an apparatus for detecting and treating a tachyarrhythmia , filed nov . 15 , 1990 and incorporated herein by reference in its entirety . however , any known cardioversion or defibrillation pulse generation circuitry is believed usable in conjunction with the present invention . for example , circuitry controlling the timing and generation of cardioversion and defibrillation pulses as disclosed in u . s . pat . no . 4 , 384 , 585 , issued to zipes on may 24 , 1983 , in u . s . pat . no . 4 , 949 , 719 issued to pless et al ., cited above , and in u . s . pat . no . 4 , 375 , 817 , issued to engle et al ., all incorporated herein by reference in their entireties may also be employed . similarly , known circuitry for controlling the timing and generation of anti - tachycardia pacing pulses as described in u . s . pat . no . 4 , 577 , 633 , issued to berkovits et al . on mar . 25 , 1986 , u . s . pat . no . 4 , 880 , 005 , issued to pless et al . on nov . 14 , 1989 , u . s . pat . no . 4 , 726 , 380 , issued to vollmann et al . on feb . 23 , 1988 and u . s . pat . no . 4 , 587 , 970 , issued to holley et al . on may 13 , 1986 , all of which are incorporated herein by reference in their entireties may also be used . in modern pacemaker / cardioverter / defibrillators , the particular anti - tachycardia and defibrillation therapies are programmed into the device ahead of time by the physician , and a menu of therapies is typically provided . for example , on initial detection of tachycardia , an anti - tachycardia pacing therapy may be selected . on re - detection of tachycardia , a more aggressive anti - tachycardia pacing therapy may be scheduled . if repeated attempts at anti - tachycardia pacing therapies fail , a higher level cardioversion pulse therapy may be selected thereafter . prior art patents illustrating such pre - set therapy menus of anti - tachyarrhythmia therapies include the above - cited u . s . pat . no . 4 , 830 , 006 , issued to haluska , et al ., u . s . pat . no . 4 , 727 , 380 , issued to vollmann et al . and u . s . pat . no . 4 , 587 , 970 , issued to holley et al . the present invention is believed practicable in conjunction with any of the known anti - tachycardia pacing and cardioversion therapies , and it is believed most likely that the invention of the present application will be practiced in conjunction with a device in which the choice and order of delivered therapies is programmable by the physician , as in current implantable pacemaker / cardioverter / defibrillators . in addition to varying the therapy delivered following a failed attempt to terminate a tachyarrhythmia , it is also known that adjustment of detection criteria may be appropriate . for example , adjustment may comprise reducing the number of intervals required to detect a tachyarrhythmia to allow a more rapid re - detection or by changing the interval ranges to bias detection towards detection of ventricular fibrillation , for example as disclosed in u . s . pat . no . 4 , 971 , 058 , issued to pless et al . and incorporated herein by reference in its entirety . in the present invention , selection of the particular electrode configuration for delivery of the cardioversion or defibrillation pulses is controlled via output circuit 548 , under control of cardioversion / defibrillation control circuitry 554 via control bus 546 . output circuit 548 determines which of the high voltage electrodes 506 , 508 and 510 will be employed in delivering the defibrillation or cardioversion pulse regimen , and may also be used to specify a multi - electrode , simultaneous pulse regimen or a multi - electrode sequential pulse regimen . monophasic or biphasic pulses may be generated . one example of circuitry which may be used to perform this function is set forth in commonly assigned co - pending patent application ser . no . 07 / 612 , 758 , filed by keimel , for an apparatus for delivering single and multiple cardioversion and defibrillation pulses , filed nov . 14 , 1990 , incorporated herein by reference in its entirety . however , output control circuitry as disclosed in u . s . pat . no . 4 , 953 , 551 , issued to mehra et al . on sep . 4 , 1990 or u . s . pat . no . 4 , 800 , 883 , issued to winstrom on jan . 31 , 1989 both incorporated herein by reference in their entireties , may also be used in the context of the present invention . alternatively single monophasic pulse regimens employing only a single electrode pair according to any of the above cited references which disclose implantable cardioverters or defibrillators may also be used . fig3 a and 3b illustrate the function of the present invention as embodied in a device as illustrated in fig2 in the form of flow charts . fig3 b illustrates the fdi adjust functional block 11 of fig3 a in more detail . fig3 a illustrates the overall tachyarrhythmia detection function as employed in the disclosed embodiment of the present invention . with the exception of functional block 11 , this portion of the tachyarrhythmia detection function corresponds to that employed in the medtronic model 7216 and 7217 implantable pacemaker / cardioverter / defibrillators , discussed above . in the context of fig3 a , the device should be understood to be operating as a demand pacer , with the detection functions illustrated taking place during the refractory period following the occurrence of a spontaneous or paced r - wave . the microprocessor waits at 10 for an interrupt indicating the occurrence of a paced or sensed r - wave and in response thereto stores the duration of the preceding r -- r interval and increments the value of vfec or vtec , if appropriate , using the interval criteria discussed above , based on the value of fdi c and tdi . the microprocessor then determines whether the value of fdi c needs to be incremented or decremented at 11 . at 12 , the microprocessor determines whether the detection criteria for ventricular fibrillation have been met , i . e ., whether vfec is greater than or equal to vfnid . if ventricular fibrillation is detected , then the scheduled defibrillation therapy is initiated in block 16 and the detection criteria and therapy menus are updated at 18 , as described above . if ventricular fibrillation is not detected at 12 , the microprocessor checks at 14 to determine whether the criteria for detection of ventricular tachycardia have been met , i . e . whether vtec is greater than or equal to vtnid . if ventricular tachycardia is detected , the scheduled ventricular tachycardia therapy is delivered at 16 and the detection criteria and therapy menus are updated at 18 , as described above . if no tachyarrhythmia is detected , if a tachycardia was previously detected , the microprocessor checks at 22 to determine whether a return to sinus rhythm has occurred , i . e . a series of a predetermined number of r -- r intervals greater than or equal to tdi . if termination is detected , the detection criteria and therapy menus are updated at 24 , as described above . the flow chart of fig3 b illustrates the method by which the value of fdi c is adjusted in block 11 of fig3 a , based on the proportion of the n most recent event intervals that are shorter than the fdi c value . decision block 40 determines whether or not n event intervals less than tdi have been stored since the last updating of the detection criteria due to detection of a tachyarrhythmia or detection of termination of a tachyarrhythmia . if not , the adjustment function is not enabled , and the microprocessor continues with the detection methodology of fig3 a . n is intended to be a relatively small number , e . g . four or eight , preferably less than vfnid or vtnid , so detection of fibrillation or tachycardia , as a practical matter will not occur until there has at least been an opportunity for the adjustment function to operate . if n intervals less than tdi have been detected , decision block 42 determines whether or not a predetermined number of the n intervals are less than the value of fdi c . if a predetermined number m of the of the n stored event intervals are less than fdi c , for example three or more of four , the microprocessor will check at 44 to determine whether fdi c is already at its maximum value . if it is at its maximum value , then it is not altered , and the present value of fdi c is employed in the detection methodology of fig3 a . however , if fdi c is not yet at its programmed maximum value , then fdi c is incremented by delta in block 46 . if m of the of the n stored event intervals are not less than fdi c , the microprocessor will check at 43 to determine whether p of the of the n stored intervals are greater than or equal to fdi c , for example three or more of four . if not , the adjustment function is not enabled , and the microprocessor continues with the detection methodology of fig3 a . if p of the n intervals are greater than or equal to fdi c , the microprocessor checks at 48 to determine whether fdi c is already at its minimum value . if it is at its minimum value , then it is not altered , and the present value of fdi c is employed in the detection methodology of fig3 a . however , if fdi c is not yet at its programmed minimum value , then fdi c is decremented by delta in block 50 . through these adjustments of the fdi c , the detection function illustrated in figure # a is rendered more sensitive to the trend of the n most recent event intervals . it is thus expected , that in practice the adjustment of the fdi c will prove beneficial in accelerating a detection of ventricular tachycardia or ventricular fibrillation in those cases in which the r -- r intervals of the patient &# 39 ; s tachyarrhythmia include intervals greater and less than fdi . after incrementing or decrementing the value of fdi c , the new value is used in subsequent detection functions illustrated in fig3 a . the new value of fdi c may be employed in various ways . the simplest manner in which the new value of fdi c may be employed is for subsequent r -- r intervals to be classified based on the new value , with the classification of preceding r -- r intervals left undisturbed . in this case , the vales of vfec and vtec would remain unaltered as a result of the adjustment function , and would simply be subsequently incremented using the new interval ranges defined using the adjusted value of fdi c . an alternative method of employing the adjusted value of fdi c is to apply the new interval ranges defined by the adjusted value both prospectively and retrospectively . in this case , previously stored r -- r intervals would be reexamined , and the vfec and vtec counts updated to the values which they would have had if the adjusted value of fdi c had been in effect throughout the detection sequence . following a completed detection sequence or detection of termination of a previously detected tachyarrhythmia , the value of fdi c is reset to be equal to fdi p , as part of the procedure for updating the detection criteria in functional blocks 18 and 24 in fig3 a . in some embodiments of the invention , the value of fdi p may be different during re - detection sequences from the value during initial detection sequences . in such cases , during re - detection sequences , the adjustment function may be employed using fdi c set to the current value of fdi p . alternatively , in such embodiments and in other embodiments of the invention , the adjustment function may be dispensed with entirely during re - detection sequences . while the preferred embodiment of the device takes the form of a microprocessor controlled device as illustrated in fig2 in which the various functional steps illustrated in fig3 a and 3b would be implemented in the form of software , the invention may equally well be practiced in the form of a dedicated , full custom digital integrated circuit or , even in the form of an analog circuit , employing analog values as substitutes for the digital values disclosed in conjunction with the above specification . in addition , while the preferred embodiment disclosed above takes the form of a pacemaker / cardioverter / defibrillator , the enhanced ability to distinguish between various tachyarrhythmias and the improved speed of detection provided by the present invention are also valuable and applicable to devices which are only capable of performing a subset of the various therapies discussed above in conjunction with fig2 . for example , the ability to accurately distinguish between ventricular tachycardia and ventricular fibrillation would be valuable in an anti - tachycardia pacemaker , even without a cardioversion pulse generator , to determine whether anti - tachycardia pacing therapies are appropriate . similarly , the ability to distinguish between ventricular tachycardia and ventricular fibrillation is valuable in an implantable cardioverter / defibrillator lacking a cardiac pacing function , for example , as in the currently available cpi aicd implantable cardioverter / defibrillators . it should further be kept in mind that while the therapies described for delivery in response to detection of the various arrhythmias discussed are all disclosed in the context of electrical therapies , it is possible that the invention may be embodied in the form of an implantable drug dispenser , wherein one or more of the anti - tachycardia therapies takes the form of injection of a drug locally into the heart or systemically to treat the detected arrhythmia . as such , the above disclosure should be taken merely as an example of an embodiment of the present invention , rather than limiting , when reading the claims which follow .

an automatic implantable device for detecting and differentiating between tachyarrhythmias in order to therapeutically stimulate the heart in response thereto , particularly for distinguishing fibrillation from tachycardia and to provide appropriate therapies for each condition . the event intervals between successive heart depolarizations are measured , stored and classified as within fibrillation or tachycardia interval ranges . the numbers of intervals falling within the fibrillation and tachycardia interval ranges are employed to distinguish fibrillation from tachycardia . the number of intervals required to detect and discriminate between tachycardia and fibrillation in situations where the tachyarrhythmia includes intervals in both interval ranges is reduced by adjusting the interval ranges as a function of the relative distribution of measured intervals within the interval ranges .

a preferred embodiment of a safety syringe 1 will be hereafter described with reference to the figures . fig1 shows a non - activated safety syringe 1 comprising a plunger 2 having a weak part 3 , a coupling portion 4 , and a distal portion 5 . the plunger sits within a syringe casing 24 . attached to the coupling portion 4 of the plunger 2 is a stopper 11 . the area within the casing 24 between a needle holder 16 and the stopper 11 defines a fluid chamber 14 which contains a fluid to be administered to a patient . fig7 shows a side view of the plunger 2 whereas fig8 focuses on the weak part 3 of the plunger 2 . on either side of the weak part 3 are the coupling portion 4 and the distal portion 5 . the coupling portion 4 has a front resistance plate 42 . the resistance plate 42 has a transitional arc brim on its front face whereas its rear face is substantially flat . the outer diameter of the resistance plate 42 is equal to or slightly smaller than the inner diameter of the syringe casing 24 . although the diameter of the resistance plate 42 can be 0 . 1 mm smaller than the inner diameter of the syringe casing 24 , it should be larger than the inner diameter of a resistance ring 37 ( hereafter described ) circumferentially formed on the inner surface of the syringe casing 24 ( as shown in fig6 ). preferably , the weak part 3 is comprised of a small number of tines 8 which are attached to and positioned between the front resistance plate 42 and the distal portion 5 . the tines 8 are designed to break easily when a bending moment is applied to the distal portion 5 when the plunger 2 is in a fully extended position . however , numerous other embodiments can be employed to create a breakable weak part 3 . the coupling portion 4 contains a plurality of hooks 6 which are designed to engage an arrowhead 7 , as described below . the distal portion 5 , on the other hand , can have a variety of cross sections . to balance cost and effectiveness , a “+” shape is preferable for the distal portion 5 ; if this configuration is used , it is preferable to have a flat plate 32 affixed to the end of the plunger 2 so that a user will have a broad flat surface on which to place a thumb when compressing the plunger 2 into the syringe casing 24 . if the “+” shape is employed to form the distal portion 5 , one plane of the distal portion 5 ( i . e . a vertical rib plate 43 ) may have tines 8 extend therefrom ( and attach to the front resistance plate 43 ) whereas the other plane of the “+” shaped distal portion 5 ( i . e . a horizontal rib plate 44 ) may have no such tines 8 . moreover , if the “+” shape is employed , in one of the sides of the cross section , a notch 9 may be positioned . the notch 9 is designed to engage a safety mechanism 10 which is preferably attached to the syringe casing 24 , as described below . as can be seen in fig1 , centrally positioned in one end of a stopper 11 is a bore 12 which leads to a cavity 45 . the bore 12 does not completely penetrate the stopper 11 . rather , the other end of the stopper 11 comprises a membrane 13 , the function of which is hereafter described . however , the bore 12 is extended by a smaller diameter bore 41 which extends to an inner side of the membrane 13 . in construction , the coupling portion 4 of the plunger 2 is completely inserted into the bore 12 of the stopper 11 until it rests in the cavity 45 . preferably , the stopper 11 is made of rubber or a rubber - like material which allows the stopper to prevent the fluid ( to be administered to a patient ) from escaping a fluid chamber 14 in the syringe 1 . in preventing the passage of such fluids , the stopper 11 has a plurality of o - rings 15 formed along its outer circumference which frictionally engage the inner surface of the fluid chamber 14 . the stopper 11 substantially covers the hooks 6 of the coupling portion 4 of the plunger 2 ; the hooks 6 are positioned in the cavity 45 and are covered by the membrane 13 prior to an activation of the syringe described below . fig5 which shows a perspective view of a needle holder 16 , depicts another o - ring 17 in a central portion of the needle holder 16 . the o - ring 17 of the needle holder 16 sits within a circumferential channel and prevents the fluid ( to be administered to a patient ) from escaping the syringe 1 through a bore 18 , described below , and also prevents accidental needle ejection when large compressive loads are applied to the plunger 2 . the o - ring 17 is designed to engage a circumferential channel 26 in the cylindrical neck portion 25 of the syringe casing 24 , ( as shown in fig2 ) as hereafter described . on one side of the o - ring 17 is a needle adapter 19 onto which a needle 20 is fixed . the needle adapter 19 is conical in shape which allows needles of various sizes to be fixed on the needle holder 16 . on the other side of the o - ring 17 is an arrowhead 7 . on one end of the arrowhead is a point 29 . at the base of the arrowhead 7 is a circumferential groove 28 designed to engage the hooks 6 of the plunger 2 , as described below . the base of the arrowhead 7 is made to be inclined , i . e . the angle of the base is at an angle a with respect to a coplanar line which is perpendicular to the axis of the syringe casing 24 ; the angle is preferably approximately 5 degrees . this angular orientation allows the needle holder 16 to be oriented at an angle when it is withdrawn into the syringe casing 24 , as described below . at the base of the groove 28 there are two flaps 40 which are designed to engage the wall 31 ( as shown in fig2 ) in the cylindrical neck 25 when the needle holder 16 is positioned in the cylindrical neck 25 . by engaging the wall 31 , the flaps 40 prevent the needle holder 16 from being ejected when the plunger 2 is compressed . moreover , the engagement of the flaps 40 and the wall 31 reduces injection residue . a needle holder axis 23 is defined by a line passing through the arrowhead 7 in the direction of the needle adapter 19 . positioned in the needle adapter 19 of the needle holder 16 is a bore 22 which travels from the needle adapter end of the needle holder 16 along the axis 23 to a position on needle holder 16 which is on the arrowhead 7 side of the o - ring 17 ; the bore 22 does not extend into the arrowhead 7 . a trans - bore 21 is perpendicularly drilled through the needle holder 16 at the end of the bore 22 , i . e . perpendicular to the axis 23 of the needle holder 16 . the two bores 21 and 22 thereby form a “ t ” shaped channel in the needle holder 16 . when a needle 20 is attached to the needle adapter 19 the “ t ” shaped channel extends into the needle . this construction allows a fluid , housed within the fluid chamber 14 , to pass through the “ t ” shaped channel ( when the plunger 2 is depressed ), through the needle 20 , and into a patient . fig6 shows a cross sectional side view of the syringe casing 24 . the casing 24 is substantially cylindrical and hollow . on one end of the casing 24 , there is a narrower cylinder 25 which extends from the casing 24 . prior to use , the cylinder 25 ( which comprises bore 18 ) holds the needle holder 16 with the arrowhead 7 distally extending into the fluid chamber 14 . in the cylinder 25 , there is a circumferential channel 26 on the inner surface of the cylinder 25 . o - ring 17 of the needle holder 16 initially rests within channel 26 and thereby prevents drugs , stored in the fluid chamber 14 , from being able to leak out of the syringe 1 . the circumferential channel 26 has an asymmetric cross - section ; the rear portion 38 has the same arc as the o - ring 17 whereas the frontal portion 39 ( i . e . the portion closest to the needle end of the syringe 1 ) has a larger arc . attached to the other end of the casing 24 are two finger supports 33 ( as shown in fig1 ) and the safety mechanism 10 . safety mechanism 10 is formed onto the casing in such a way as to allow it to swing toward and away from the plunger 2 when the plunger 2 is positioned within the casing 24 . to allow the safety mechanism 10 to swing , the safety mechanism is comprised of a wing plate 34 ( which is a part of the finger support 33 ) which is connected to a check plate 35 by means of a thin plastic film 36 . preferably , the check plate 35 has a notch 27 ( as shown in fig1 ) which is sized to correspond to the thickness of one of the sides of the “+” shaped plunger . when the check plate 35 is rotated from an unlocked position and toward the plunger 2 and thereby into a locked position , the notch 27 of the check plate 35 may engage the notch 9 in one of the sides of the “+” shaped plunger 2 . a user will be unable to activate ( as described below ) the syringe 1 by depressing the plunger 2 while the notch 27 is engaged with notch 9 as this engagement prevents further compression of the plunger 2 ; accordingly , completely activation of the syringe 1 is avoided . in addition , the rear end of the syringe casing 24 also comprises a resistance ring 37 formed on the interior of the casing 24 near the end of the casing 24 in which the plunger is inserted . the resistance ring 37 has a cross - section substantially triangular in shape . the cross - section of the ring 37 is such that the portion 47 of the cross - section facing the needle end of the syringe 1 is steeper than the portion 48 facing the plunger 2 end of the syringe 1 . in this fashion , the portion 47 of the ring 37 facing the needle 20 end of the syringe 1 has a larger axial angle with respect to the axis of the syringe casing 24 ; the angle is preferably about 60 degrees . the side of the ring 37 facing the plunger 2 end of the syringe 1 has a smaller axial angle with respect to the axis of the syringe casing 24 ; the angle is preferably about 10 degrees . during assembly , the needle holder 16 is channeled into the syringe casing 24 through the plunger 2 end of the casing 24 . the needle holder 16 is then pushed forward until the o - ring 17 snaps into the circumferential channel 26 and the flaps 40 hit a circumferential rim 46 located in the cylindrical neck portion 25 adjacent the circumferential wall 31 . the hooks 6 of the plunger 2 are inserted into the bore 12 in the stopper 11 until the are housed in the cavity 45 ; the combination of the plunger 2 and the stopper 11 is then inserted into the syringe casing 24 . in so doing , the front resistance plate 42 of the plunger 2 rides over the resistance ring 37 to become permanently housed with the syringe casing 24 . the check plate 35 of the safety mechanism 10 should be turned down toward the plunger 2 ( i . e . into the locked position ) by means of the bendable film plastic film 36 and its notch 27 should be aligned with the side of the “+” shaped plunger containing the corresponding notch 9 so that an engagement of the notches 9 , 27 can occur thereby preventing complete compression of the plunger 2 in the syringe casing 24 . activation of the syringe 1 occurs as follows . when the safety mechanism 10 is not engaged with one of the sides of the plunger 2 and a user depresses the plunger 2 to compress the fluid chamber 14 , drugs in the fluid chamber are forced into the “ t ” shaped channel of the needle holder 16 and into ( and out of ) the needle 20 . as the plunger is compressed , the membrane 13 of the stopper 11 approaches the arrowhead 7 . with continued compression , the point 29 of the arrowhead 7 pierces the membrane 13 causing the arrowhead 7 to pass through bore 41 and into cavity 45 in which the arrowhead 7 engages the hooks 6 . again , with continued compression , the arrowhead 7 radially separates the hooks 6 to a point at which the hooks 6 snap into the circumferential groove 28 around the base of the arrowhead 7 as shown in fig1 . in this fashion the arrowhead 7 and the plunger 2 become permanently engaged within the stopper 11 . due to the engagement of the hooks 6 with the circumferential groove 28 , when a user pulls on the plunger 2 , the needle holder 16 ( to which the arrowhead 7 is attached ) and the needle 20 are pulled into the syringe casing 24 . the user is unable to pull the plunger ( with needle attached ) out of the syringe casing 24 because front resistance plate 42 of the plunger 2 engages the frontal portion 47 of the resistance ring 37 . as shown in fig4 when the needle 20 is completely pulled into the syringe casing 24 , the distal portion 5 of the plunger 2 extends well out of the casing 24 . with a small bending moment applied to a side of the distal portion 5 , the tines 8 of the weak point 3 break . as shown in fig4 and 9 , when the tines 8 break , the distal portion 5 of the plunger 2 becomes disengaged from the coupling portion 4 of the plunger 2 . without the distal portion 5 attached to the coupling portion 4 of the plunger 2 , a user is unable to push the needle 20 back out of the casing 24 . if , on the other hand , the user does not break the distal portion 5 and instead tries to push the needle 20 out of the casing 24 , the user will be unable to push the needle 20 out of the casing 24 for the reason discussed below . as shown in fig2 when the needle holder 16 is drawn into the casing 24 ( after complete activation ) and the skin contact end 30 of the needle 20 clears the bore 18 of the cylinder 25 , the skin contact end 30 of the needle 20 is forced toward the inner surface of the casing 24 . the needle 20 is tipped toward the inner surface of the casing 24 because the arrowhead 7 and hooks 6 are engaged at an angle of approximately 5 degrees ; the angular orientation is caused by the inclined base of the arrowhead 7 as previously described . in this fashion , the axis defined by the needle 20 ( when completely within the casing 24 ) is not parallel to the axis upon which the plunger 2 is pulled by the user . as shown in fig3 if a user tries to push the needle 20 back out of the casing 24 , the skin contact end 30 of the needle 20 does not pass through bore 18 but rather contacts a wall 31 of the casing which extends circumferentially from the outer surface of the casing 24 toward the cylinder 25 . due to the contact between the skin contact end 30 of the needle 20 and the wall 31 , the needle is unable to exit the casing 24 . moreover , if the user continuously increases the force applied to the plunger 2 , the needle will collapse . although the aforementioned described a preferred embodiment of the invention , the invention is not so restricted . the foregoing description is for exemplary purposes only and is not intended to be limiting . accordingly , alternatives which would be obvious to one of ordinary skill in the art upon reading the teachings herein disclosed , are hereby within the scope of this invention . the invention is limited only as defined in the following claims and equivalents thereof .

a safety syringe comprising a casing having a neck and a plunger having a distal portion , a weak portion , and a coupling portion covered by a hollow stopper having a membrane . the plunger is channeled in the casing and is moveable between an extended position and a compressed position . a needle holder , in the neck , contains a distally facing arrowhead and a proximally facing needle . moving the plunger from an extended position to a compressed position , causes the arrowhead to puncture the membrane and engage the coupling portion . if the plunger is returned to the extended position , the needle is drawn into the casing and the distal portion of the plunger can be removed . if the plunger is re - compressed , the needle encounters a wall in the neck preventing the needle from exiting the casing . the syringe is easy to manufacture and prevents manufacturing burrs from being injected into patients .

referring now to the drawings , fig1 and 2 , respectively , show comparative views of a prior art panty and a panty according to the present invention . as shown in fig1 the prior art panty allows a wearer &# 39 ; s buttocks to droop , making an unappealing appearance . in contrast , fig2 shows an environmental view of a panty 10 according to the present invention . as clearly shown in the drawing , the panty 10 lifts and separates the cheeks of a wearer &# 39 ; s buttocks , giving a more shapely and youthful appearance to the wearer &# 39 ; s body . fig3 shows a front view of a panty 10 according to the invention . the panty 10 includes a front panel 14 and a rear panel 16 . the front panel 14 can assume any of a variety of conventional configurations , and can be specially configured to accommodate either female or male anatomy . the panty 10 desirably includes a waistband 12 for securing the panty about a wearer &# 39 ; s waist , which can be made in the form of an integrally knitted turned welt or as an attached piece of elastic banding , for example . the panty 10 includes first and second leg openings 18 which define a crotch portion 20 therebetween . the crotch portion 20 can be formed in any conventional manner , and desirably includes a pile surface for contacting the wearer &# 39 ; s body . in a preferred embodiment of the invention , elastic banding 22 is provided around each of the leg openings 18 so that the banding reduces the diameter of the leg openings 18 . in this way , the leg openings can readily expand to allow a wearer &# 39 ; s legs to enter the leg openings 18 and contract to securely grip the legs and maintain the panty in its proper position on the wearer &# 39 ; s body . the front panel 14 is joined to the rear panel 16 at a seam 24 adjacent crotch portion 22 , and along first and second sides 26 . preferably , seam 24 is a sewn seam and the front and rear panels are joined at sides 26 by knitting them integrally . as shown in fig4 the panty 10 includes an integrally knit support panel 30 on its rear panel 16 . the support panel 30 is substantially anchor - shaped , and includes a vertically extending strip 32 and portions 34 which curve outwardly from the strip 32 toward and along a portion of said first and second leg openings 18 , to thereby conform to those regions of a wearer &# 39 ; s body corresponding to the areas between and beneath a wearer &# 39 ; s buttock cheeks . the support panel 30 is formed so as to have greater resistance to stretch , particularly in the walewise direction , than the remainder 28 of the rear panel 16 . in this way , the curved portions 34 tuck underneath a wearer &# 39 ; s buttocks while vertically extending strip 32 cooperates with the curved portions 34 to lift and separate the individual buttock cheeks . the support panel 30 can be made to extend from underneath the cheeks to the seam 24 joining the front and rear panels 14 and 16 , as shown in the embodiment in fig4 . alternatively , the curved portions 34 of the support panel 30 can assume a strip - like configuration , as shown in the embodiment of fig5 . the embodiment of fig5 depicts the curved portions 34 having a thickness similar to that of the vertically extending strip 32 . however , it is noted that the curved portions 34 could be thicker than the vertically extending strip 32 , while still not extending all the way to the seam 24 joining the front and rear panels 14 and 16 . as shown in the embodiment in fig6 the support panel 30 can be made of individual spaced regions 36 , provided the overall effect is to form a substantially anchor - shaped support panel which conforms to the regions corresponding to those regions of a wearer &# 39 ; s body corresponding to the areas between and beneath a wearer &# 39 ; s buttock cheeks . fig7 and 8 depict rear and front views , respectively , of a blank for forming a panty according to the invention . the tubular blank , shown generally at 40 , preferably includes a turned welt portion about its upper end . the tubular blank includes a first side 44 for forming the rear panel 16 of a panty . the first side 44 is integrally knit with second side 52 , which forms the front panel 14 of a panty . the first side 44 includes a substantially anchor - shaped region 46 which is knitted so as to be more resistant to stretch than the remainder of the first side 44 of the blank 40 . this substantially anchor - shaped region 46 of the blank 40 forms the support panel 30 of the completed panty 10 , and includes a walewise extending strip 48 and two upwardly curved portions 50 which extend outwardly from a lower portion of the walewise extending strip 48 . as shown in fig8 the second side 52 of the blank 40 preferably includes a region 54 having a modified knit configuration for forming the crotch portion 20 of the panty 10 . a particularly preferred knit configuration is a pile - forming configuration , for forming a pile surface on the wearer - contacting surface of the crotch portion 20 . the manufacture of a panty according to the present invention is performed as follows : a series of courses is knitted on a circular knitting machine to form a tubular blank 40 having a substantially anchor - shaped region 46 which is more resistant to stretch than the remainder of the tubular blank and integrally formed with the tubular blank . this is preferably done by floating in an extra elastic yarn in the more stretch resistant region or by forming this region using a pucker stitch , wherein yarns are held during the formation of selected portions of selected alternating courses . however , other conventional methods for forming this region so that it is more stretch resistant than the rest of the blank can be employed . the substantially anchor - shaped region 46 includes a walewise extending strip 48 and two upwardly curved portions 50 extending outwardly from a lower portion of the walewise extending strip 48 . in addition , the tubular blank 40 is also desirably knit to have a turned welt 42 about its upper end , at a position remote from the upwardly curved portions 50 of the substantially anchor - shaped region 46 . in a preferred embodiment of the invention , the tubular blank 40 also includes a region 54 on the side of the blank 40 opposite that having the substantially anchor - shaped region 46 . the region 54 has a pile forming knit configuration which forms the crotch portion of the panty . the tubular blank 40 is then cut and first and second spaced areas of the blank are removed to form first and second leg openings and define a crotch portion therebetween and front and rear panty panels 14 and 16 . the front and rear panty panels 14 and 16 are then joined together between the leg openings 18 at the seam 24 , which preferably is a sewn seam . in addition , elastic banding 22 is desirably attached around the leg openings 18 , to reduce the diameter of the openings . however , it is noted that the front and rear panels 14 and 16 can be attached together at seam 24 prior to cutting and removing the first and spaced areas to form the leg openings . in the drawings and the specification , there has been set forth a preferred embodiment of the invention , and although specific terms are employed , they are used in a generic and descriptive sense only and not for purposes of limitation , the scope of the invention being defined in the claims .

the invention discloses a panty - type undergarment having a support panel which has a greater resistance to stretch than the rest of the undergarment for lifting and separating the cheeks of a wearer &# 39 ; s buttocks . the support panel is substantially anchor - shaped and includes a vertical strip and upwardly curving portions which extend toward and along a portion of the undergarment leg openings . the undergarment is circularly knit and includes modified knit regions to form the portions forming the support panel . a method and blank for making the panty with the lift and separation support are also disclosed .

in one embodiment , there are provided compounds having the structure ( i ): r 1 to r 3 are each independently — h , alkyl , substituted alkyl , alkenyl , substituted alkenyl , alkynyl , substituted alkynyl , aryl , substituted aryl , heteroaryl , substituted heteroaryl , heterocyclic , substituted heterocyclic , cycloalkyl , substituted cycloalkyl , alkoxy , substituted alkoxy , thioalkyl , substituted thioalkyl , hydroxy , halogen , amino , amido , carboxyl , — c ( o ) h , acyl , oxyacyl , carbamate , sulfonyl , sulfonamide , or sulfuryl ; each r 4 is independently alkyl , substituted alkyl , alkenyl , substituted alkenyl , alkynyl , substituted alkynyl , aryl , substituted aryl , cycloalkyl , substituted cycloalkyl ; e 1 to e 4 are each independently — o , — nr 5 , or — s , wherein r 5 is — h or c 1 - c 6 alkyl ; and x is 0 to 8 . in a further embodiment of the invention , there are provided compounds having the structure ( ii ): r 1 to r 3 are each independently — h , alkyl , substituted alkyl , alkenyl , substituted alkenyl , alkynyl , substituted alkynyl , aryl , substituted aryl , heteroaryl , substituted heteroaryl , heterocyclic , substituted heterocyclic , cycloalkyl , substituted cycloalkyl , alkoxy , substituted alkoxy , thioalkyl , substituted thioalkyl , hydroxy , halogen , amino , amido , carboxyl , — c ( o ) h , acyl , oxyacyl , carbamate , sulfonyl , sulfonamide , or sulfuryl ; each r 4 is independently alkyl , substituted alkyl , alkenyl , substituted alkenyl , alkynyl , substituted alkynyl , aryl , substituted aryl , cycloalkyl , substituted cycloalkyl ; e 1 to e 4 are each independently — o , — nr 5 , or — s , wherein r 5 is — h or c 1 - c 6 alkyl ; and x is 0 to 8 . in one embodiment , there are provided compounds having the structure ( iii ): r 1 to r 3 are each independently — h , alkyl , substituted alkyl , alkenyl , substituted alkenyl , alkynyl , substituted alkynyl , aryl , substituted aryl , heteroaryl , substituted heteroaryl , heterocyclic , substituted heterocyclic , cycloalkyl , substituted cycloalkyl , alkoxy , substituted alkoxy , thioalkyl , substituted thioalkyl , hydroxy , halogen , amino , amido , carboxyl , — c ( o ) h , acyl , oxyacyl , carbamate , sulfonyl , sulfonamide , or sulfuryl , each r 4 is independently alkyl , substituted alkyl , alkenyl , substituted alkenyl , alkynyl , substituted alkynyl , aryl , substituted aryl , cycloalkyl , substituted cycloalkyl , e 1 to e 4 are each independently — o , — nr 5 , or — s , wherein r 5 is — h or c 1 - c 6 alkyl , and x is 0 to 8 . in still a further embodiment of the invention , there are provided compounds having the structure ( iv ): in a further embodiment of the invention , there are provided compounds having the structure ( v ): in a further embodiment of the invention , there are provided compounds having the structure ( vi ): as used herein , the term “ alkyl ” refers to a monovalent straight or branched chain hydrocarbon group having from one to about 12 carbon atoms , including methyl , ethyl , n - propyl , isopropyl , n - butyl , isobutyl , tert - butyl , n - hexyl , and the like . as used herein , “ substituted alkyl ” refers to alkyl groups further bearing one or more substituents selected from hydroxy , alkoxy , mercapto , cycloalkyl , substituted cycloalkyl , heterocyclic , substituted heterocyclic , aryl , substituted aryl , heteroaryl , substituted heteroaryl , aryloxy , substituted aryloxy , halogen , cyano , nitro , amino , amido , — c ( o ) h , acyl , oxyacyl , carboxyl , sulfonyl , sulfonamide , sulfuryl , and the like . as used herein , “ lower alkyl ” refers to alkyl groups having from 1 to about 6 carbon atoms . as used herein , “ alkenyl ” refers to straight or branched chain hydrocarbyl groups having one or more carbon - carbon double bonds , and having in the range of about 2 up to 12 carbon atoms , and “ substituted alkenyl ” refers to alkenyl groups further bearing one or more substituents as set forth above . as used herein , “ alkynyl ” refers to straight or branched chain hydrocarbyl groups having at least one carbon - carbon triple bond , and having in the range of about 2 up to 12 carbon atoms , and “ substituted alkynyl ” refers to alkynyl groups further bearing one or more substituents as set forth above . as used herein , “ aryl ” refers to aromatic groups having in the range of 6 up to 14 carbon atoms and “ substituted aryl ” refers to aryl groups further bearing one or more substituents as set forth above . as used herein , “ heteroaryl ” refers to aromatic rings containing one or more heteroatoms ( e . g ., n , o , s , or the like ) as part of the ring structure , and having in the range of 3 up to 14 carbon atoms and “ substituted heteroaryl ” refers to heteroaryl groups further bearing one or more substituents as set forth above . as used herein , “ alkoxy ” refers to the moiety — o - alkyl -, wherein alkyl is as defined above , and “ substituted alkoxy ” refers to alkoxyl groups further bearing one or more substituents as set forth above . as used herein , “ thioalkyl ” refers to the moiety — s - alkyl -, wherein alkyl is as defined above , and “ substituted thioalkyl ” refers to thioalkyl groups further bearing one or more substituents as set forth above . as used herein , “ cycloalkyl ” refers to ring - containing alkyl groups containing in the range of about 3 up to 8 carbon atoms , and “ substituted cycloalkyl ” refers to cycloalkyl groups further bearing one or more substituents as set forth above . as used herein , “ heterocyclic ”, refers to cyclic ( i . e ., ring - containing ) groups containing one or more heteroatoms ( e . g ., n , o , s , or the like ) as part of the ring structure , and having in the range of 3 up to 14 carbon atoms and “ substituted heterocyclic ” refers to heterocyclic groups further bearing one or more substituents as set forth above . in certain embodiments , there are provided compounds of structures i - iii wherein e 1 , e 3 , and e 4 are — o , and e 2 is — nh . in certain embodiments , there are provided compounds of structures i - iii wherein r 1 and r 2 are — h , alkyl , or substituted alkyl , and r 3 is hydroxy or alkoxy . in some embodiments , r 1 is substituted alkyl . exemplary substituted alkyls contemplated for use include halogenated alkyls , such as for example chlorinated alkyls . the compounds of the invention may be formulated into pharmaceutical compositions as natural or salt forms . pharmaceutically acceptable non - toxic salts include the base addition salts ( formed with free carboxyl or other anionic groups ) which may be derived from inorganic bases such as , for example , sodium , potassium , ammonium , calcium , or ferric hydroxides , and such organic bases as isopropylamine , trimethylamine , 2 - ethylamino - ethanol , histidine , procaine , and the like . such salts may also be formed as acid addition salts with any free cationic groups and will generally be formed with inorganic acids such as , for example , hydrochloric , sulfuric , or phosphoric acids , or organic acids such as acetic , p - toluenesulfonic , methanesulfonic acid , oxalic , tartaric , mandelic , and the like . salts of the invention include amine salts formed by the protonation of an amino group with inorganic acids such as hydrochloric acid , hydrobromic acid , hydroiodic acid , sulfuric acid , phosphoric acid , and the like . salts of the invention also include amine salts formed by the protonation of an amino group with suitable organic acids , such as p - toluenesulfonic acid , acetic acid , and the like . additional excipients which are contemplated for use in the practice of the present invention are those available to those of ordinary skill in the art , for example , those found in the united states pharmacopeia vol . xxii and national formulary vol . xvii , u . s . pharmacopeia convention , inc ., rockville , md . ( 1989 ), the relevant contents of which is incorporated herein by reference . the compounds according to this invention may contain one or more asymmetric carbon atoms and thus occur as racemates and racemic mixtures , single enantiomers , diastereomeric mixtures and individual diastereomers . the term “ stereoisomer ” refers to chemical compounds which differ from each other only in the way that the different groups in the molecules are oriented in space . stereoisomers have the same molecular weight , chemical composition , and constitution as another , but with the atoms grouped differently . that is , certain identical chemical moieties are at different orientations in space and , therefore , when pure , have the ability to rotate the plane of polarized light . however , some pure stereoisomers may have an optical rotation that is so slight that it is undetectable with present instrumentation . all such isomeric forms of these compounds are expressly included in the present invention . each stereogenic carbon may be of r or s configuration . although the specific compounds exemplified in this application may be depicted in a particular configuration , compounds having either the opposite stereochemistry at any given chiral center or mixtures thereof are also envisioned . when chiral centers are found in the derivatives of this invention , it is to be understood that this invention encompasses all possible stereoisomers . the terms “ optically pure compound ” or “ optically pure isomer ” refers to a single stereoisomer of a chiral compound regardless of the configuration of the compound . salinosporamide a exhibits a molecular structure having a variety of functional groups ( lactone , alkylhalide , amide , hydroxide ) that can be chemically modified to produce synthetic derivatives . accordingly , exemplary invention compound salinosporamide a provides an excellent lead structure for the development of synthetic and semisynthetic derivatives . indeed , salinosporamide a can be derivatized to improve pharmacokinetic and pharmacodynamic properties , which facilitate administration and increase utility of the derivatives as anti - neoplastic agents . procedures for chemically modifying invention salinosporamide compounds to produce additional compounds within the scope of the present invention are available to those of ordinary skill in the art . salinosporamide a shows strong cytotoxic activity against human colon cancer cells in the htc - 116 cell assays . the ic 50 of 11 ng / ml exceeds the activity of etoposide ( see fig3 , ic 50 828 ng / ml ), an anticancer drug used for treatment of a number of cancers , by almost two orders of magnitude ( see fig4 ). this high activity makes invention salinosporamides excellent candidates for use in the treatment of various human cancers , especially slow growing , refractile cancers for which there are no therapies . salinosporamide a is specific to inhibition of mammalian cells and shows little anifungal activity against candida albicans ( ic 50 250 μg / ml ) and no antibacterial activity ( staphylococcus aureus , enterococcus faecium ). the ic 50 of salinosporamide a is far lower than the strongest chemotherapeutic agents currently in use or in clinical trials . salinosporamide a is a fermentation product of the marine actinomycete strains cnb392 and cnb476 . these strains are members of the order actinomycetales , which are high g + c gram positive bacteria . the novelty of cnb392 and cnb476 is at the genus level . invention compounds set forth herein are produced by certain “ salinospora ” sp . in some embodiments , invention compounds are produced by “ salinospora ” sp . strains cnb392 and cnb476 . to that end , the cnb476 strains of “ salinospora ” sp . were deposited on jun . 20 , 2003 , pursuant to the budapest treaty on the international deposit of microorganisms for the purposes of patent procedure with the patent culture depository of the american type culture collection , 12301 parklawn drive , rockville , md . 20852 u . s . a . under atcc accession no pta - 5275 . as is the case with other organisms , the characteristics of “ salinospora ” sp . are subject to variation . for example , recombinants , variants , or mutants of the specified strain may be obtained by treatment with various known physical and chemical mutagens , such as ultraviolet ray , x - rays , gamma rays , and n - methyl - n ′- nitro - n - nitrosoguanidine . all natural and induced variants , mutants , and recombinants of the specified strain which retain the characteristic of producing a compound of the invention are intended to be within the scope of the claimed invention . invention compounds can be prepared , for example , by bacterial fermentation , which generates the compounds in sufficient amounts for pharmaceutical drug development and for clinical trials . in some embodiments , invention compounds are produced by fermentation of the actinomycete strains cnb392 and cnb476 in a1bfe + c or cka - liquid media . essential trace elements which are necessary for the growth and development of the culture should also be included in the culture medium . such trace elements commonly occur as impurities in other constituents of the medium in amounts sufficient to meet the growth requirements of the organisms . it may be desirable to add small amounts ( i . e . 0 . 2 ml / l ) of an antifoam agent such as polypropylene glycol ( m . w . about 2000 ) to large scale cultivation media if foaming becomes a problem . the organic metabolites are isolated by adsorption onto an amberlite xad - 16 resin . for example , salinosporamide a is isolated by elution of the xad - 16 resin with methanol : dichlormethane 1 : 1 , which affords about 105 mg crude extract per liter of culture . salinosporamide a is then isolated from the crude extract by reversed - phase flash chromatography followed by reverse - phase hplc and normal phase hplc , which yields 6 . 7 mg of salinosporamide a . fig5 sets forth a block diagram outlining isolation and separation protocols for invention compounds . the structure of salinosporamide a was elucidated by a variety of nmr techniques , mass spectroscopy , ir , and uv spectroscopy , as set forth in fig6 - 14 . the absolute structure of salinosporamide a , and confirmation of the overall structure of salinosporamide a , was achieved by single - crystal x - ray diffraction analysis ( see example 3 ). the present invention also provides articles of manufacture including packaging material and a pharmaceutical composition contained within the packaging material , wherein the packaging material comprises a label which indicates that the pharmaceutical composition can be used for treatment of disorders and wherein the pharmaceutical composition includes a compound according to the present invention . thus , in one aspect , the invention provides a pharmaceutical composition including a compound of the invention , wherein the compound is present in a concentration effective to treat cell proliferative disorders . the concentration can be determined by one of skill in the art according to standard treatment regimen or as determined by an in vivo animal assay , for example . pharmaceutical compositions employed as a component of invention articles of manufacture can be used in the form of a solid , a solution , an emulsion , a dispersion , a micelle , a liposome , and the like , wherein the resulting composition contains one or more invention compounds as an active ingredient , in admixture with an organic or inorganic carrier or excipient suitable for enteral or parenteral applications . compounds employed for use as a component of invention articles of manufacture may be combined , for example , with the usual non - toxic , pharmaceutically acceptable carriers for tablets , pellets , capsules , suppositories , solutions , emulsions , suspensions , and any other form suitable for use . the carriers which can be used include glucose , lactose , gum acacia , gelatin , mannitol , starch paste , magnesium trisilicate , talc , corn starch , keratin , colloidal silica , potato starch , urea , medium chain length triglycerides , dextrans , and other carriers suitable for use in manufacturing preparations , in solid , semisolid , or liquid form . in addition auxiliary , stabilizing , thickening and coloring agents and perfumes may be used . the compositions of the present invention may contain other therapeutic agents as described below , and may be formulated , for example , by employing conventional solid or liquid vehicles or diluents , as well as pharmaceutical additives of a type appropriate to the mode of desired administration ( for example , excipients , binders , preservatives , stabilizers , flavors , etc .) according to techniques such as those well known in the art of pharmaceutical formulation . invention pharmaceutical compositions may be administered by any suitable means , for example , orally , such as in the form of tablets , capsules , granules or powders ; sublingually ; buccally ; parenterally , such as by subcutaneous , intravenous , intramuscular , or intracisternal injection or infusion techniques ( e . g ., as sterile injectable aqueous or non - aqueous solutions or suspensions ); nasally such as by inhalation spray ; topically , such as in the form of a cream or ointment ; or rectally such as in the form of suppositories ; in dosage unit formulations containing non - toxic , pharmaceutically acceptable vehicles or diluents . invention compounds may , for example , be administered in a form suitable for immediate release or extended release . immediate release or extended release may be achieved by the use of suitable pharmaceutical compositions comprising invention compounds , or , particularly in the case of extended release , by the use of devices such as subcutaneous implants or osmotic pumps . invention compounds may also be administered liposomally . the invention further provides methods for using invention salinosporamide compounds of structures ( i )-( vi ) to inhibit the proliferation of mammalian cells by contacting these cells with an invention salinosporamide compound in an amount sufficient to inhibit the proliferation of the mammalian cell . one embodiment is a method to inhibit the proliferation of hyperproliferative mammalian cells . for purposes of this invention , “ hyperproliferative mammalian cells ” are mammalian cells which are not subject to the characteristic limitations of growth , e . g ., programmed cell death ( apoptosis ). a further preferred embodiment is when the mammalian cell is human . the invention further provides contacting the mammalian cell with at least one invention salinosporamide compound and at least one additional anti - neoplastic agent . in another embodiment , there are provided methods for treating a mammalian cell proliferative disorder , comprising administering to a subject in need thereof a therapeutically effective amount of a compound of structures ( i )-( vi ). cell proliferative disorders that can be effectively treated by the methods of the invention include disorders characterized by the formation of neoplasms . as such , invention compounds are anti - neoplastic agents . as used herein , “ neoplastic ” pertains to a neoplasm , which is an abnormal growth , such growth occurring because of a proliferation of cells not subject to the usual limitations of growth . as used herein , “ anti - neoplastic agent ” is any compound , composition , admixture , co - mixture or blend which inhibits , eliminates , retards or reverses the neoplastic phenotype of a cell . in certain embodiments , the neoplasms are selected from mammory , small - cell lung , non - small - cell lung , colorectal , leukemia , melanoma , pancreatic adenocarcinoma , central nervous system ( cns ), ovarian , prostate , sarcoma of soft tissue or bone , head and neck , gastric which includes thyroid and non - hodgkin &# 39 ; s disease , stomach , myeloma , bladder , renal , neuroendocrine which includes thyroid and non - hodgkin &# 39 ; s disease and hodgkin &# 39 ; s disease neoplasms . in one embodiment , the neoplasms are colorectal . chemotherapy , surgery , radiation therapy , therapy with biologic response modifiers , and immunotherapy are currently used in the treatment of cancer . each mode of therapy has specific indications which are known to those of ordinary skill in the art , and one or all may be employed in an attempt to achieve total destruction of neoplastic cells . chemotherapy utilizing one or more invention salinosporamide compounds is provided by the present invention . moreover , combination chemotherapy , chemotherapy utilizing invention salinosporamide compounds in combination with other neoplastic agents , is also provided by the invention as combination therapy is generally more effective than the use of single anti - neoplastic agents . thus , a further aspect of the present invention provides compositions containing a therapeutically effective amount of at least one invention salinosporamide compound in combination with at least one other anti - neoplastic agent . such compositions can also be provided together with physiologically tolerable liquid , gel or solid carriers , diluents , adjuvants and excipients . such carriers , diluents , adjuvants and excipients may be found in the united states pharmacopeia vol . xxii and national formulary vol xvii , u . s . pharmacopeia convention , inc ., rockville , md . ( 1989 ), the contents of which are herein incorporated by reference . additional modes of treatment are provided in ahfs drug information , 1993 ed . by the american hospital formulary service , pp . 522 - 660 , the contents of which are herein incorporated by reference . anti - neoplastic agents which may be utilized in combination with an invention salinosporamide compound include those provided in the merck index , 11th ed . merck & amp ; co ., inc . ( 1989 ) pp . ther 16 - 17 , the contents of which are hereby incorporated by reference . in a further embodiment of the invention , anti - neoplastic agents may be antimetabolites which may include , but are not limited to , methotrexate , 5 - fluorouracil , 6 - mercaptopurine , cytosine arabinoside , hydroxyurea , and 2 - chlorodeoxyadenosine . in another embodiment of the present invention , the anti - neoplastic agents contemplated are alkylating agents which may include , but are not limited to , cyclophosphamide , melphalan , busulfan , paraplatin , chlorambucil , and nitrogen mustard . in a further embodiment of the invention , the antineoplastic agents are plant alkaloids which may include , but are not limited to , vincristine , vinblastine , taxol , and etoposide . in a further embodiment of the invention , the anti - neoplastic agents contemplated are antibiotics which may include , but are not limited to , doxorubicin ( adriamycin ), daunorubicin , mitomycin c , and bleomycin . in a further embodiment of the invention , the anti - neoplastic agents contemplated are hormones which may include , but are not limited to , calusterone , diomostavolone , propionate , epitiostanol , mepitiostane , testolactone , tamoxifen , polyestradiol phosphate , megesterol acetate , flutamide , nilutamide , and trilotane . in a further embodiment of the invention , the anti - neoplastic agents contemplated include enzymes which may include , but are not limited to , l - asparaginase or aminoacridine derivatives which may include , but are not limited to , amsacrine . additional anti - neoplastic agents include those provided in skeel , roland t ., “ antineoplastic drugs and biologic response modifier : classification , use and toxicity of clinically useful agents ,” handbook of cancer chemotherapy ( 3rd ed . ), little brown & amp ; co . ( 1991 ), the contents of which are herein incorporated by reference . in addition to primates , such as humans , a variety of other mammals can be treated according to the method of the present invention . for instance , mammals including , but not limited to , cows , sheep , goats , horses , dogs , cats , guinea pigs , rats or other bovine , ovine , equine , canine , feline , rodent or murine species can be treated . the term “ therapeutically effective amount ” means the amount of the subject compound that will elicit the biological or medical response of a tissue , system , animal or human that is being sought by the researcher , veterinarian , medical doctor or other clinician , e . g ., lessening of the effects / symptoms of cell proliferative disorders . by “ pharmaceutically acceptable ” it is meant the carrier , diluent or excipient must be compatible with the other ingredients of the formulation and not deleterious to the recipient thereof . the terms “ administration of ” and or “ administering a ” compound should be understood to mean providing a compound of the invention to the individual in need of treatment . administration of the invention compounds can be prior to , simultaneously with , or after administration of another therapeutic agent or other anti - neoplastic agent . the pharmaceutical compositions for the administration of the compounds of this invention may conveniently be presented in dosage unit form and may be prepared by any of the methods well known in the art of pharmacy . all methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients . in general , the pharmaceutical compositions are prepared by uniformly and intimately bringing the active ingredient into association with a liquid carrier or a finely divided solid carrier or both , and then , if necessary , shaping the product into the desired formulation . in the pharmaceutical composition the active object compound is included in an amount sufficient to produce the desired effect upon the process or condition of diseases . the pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use , for example , as tablets , troches , lozenges , aqueous or oily suspensions , dispersible powders or granules , emulsions , hard or soft capsules , or syrups or elixirs . compositions intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such compositions may contain one or more agents selected from the group consisting of sweetening agents , flavoring agents , coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations . tablets contain the active ingredient in admixture with non - toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets . these excipients may be for example , inert diluents , such as calcium carbonate , sodium carbonate , lactose , calcium phosphate or sodium phosphate ; granulating and disintegrating agents , for example , corn starch , or alginic acid ; binding agents , for example starch , gelatin or acacia , and lubricating agents , for example magnesium stearate , stearic acid or talc . the tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period . for example , a time delay material such as glyceryl monostearate or glyceryl distearate may be employed . they may also be coated to form osmotic therapeutic tablets for control release . formulations for oral use may also be presented as hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent , for example , calcium carbonate , calcium phosphate or kaolin , or as soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium , for example peanut oil , liquid paraffin , or olive oil . aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions . such excipients are suspending agents , for example sodium carboxymethylcellulose , methylcellulose , hydroxy - propylmethylcellulose , sodium alginate , polyvinyl - pyrrolidone , gum tragacanth and gum acacia ; dispersing or wetting agents may be a naturally - occurring phosphatide , for example lecithin , or condensation products of an alkylene oxide with fatty acids , for example polyoxyethylene stearate , or condensation products of ethylene oxide with long chain aliphatic alcohols , for example heptadecaethyleneoxycetanol , or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate , or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides , for example polyethylene sorbitan monooleate . the aqueous suspensions may also contain one or more preservatives , for example ethyl , or n - propyl , p - hydroxybenzoate , one or more coloring agents , one or more flavoring agents , and one or more sweetening agents , such as sucrose or saccharin . oily suspensions may be formulated by suspending the active ingredient in a vegetable oil , for example arachis oil , olive oil , sesame oil or coconut oil , or in a mineral oil such as liquid paraffin . the oily suspensions may contain a thickening agent , for example beeswax , hard paraffin or cetyl alcohol . sweetening agents such as those set forth above , and flavoring agents may be added to provide a palatable oral preparation . these compositions may be preserved by the addition of an anti - oxidant such as ascorbic acid . dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent , suspending agent and one or more preservatives . suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above . additional excipients , for example sweetening , flavoring and coloring agents , may also be present . syrups and elixirs may be formulated with sweetening agents , for example glycerol , propylene glycol , sorbitol or sucrose . such formulations may also contain a demulcent , a preservative and flavoring and coloring agents . the pharmaceutical compositions may be in the form of a sterile injectable aqueous or oleagenous suspension . this suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above . the sterile injectable preparation may also be a sterile injectable solution or suspension in a non - toxic parenterally - acceptable diluent or solvent , for example as a solution in 1 , 3 - butane diol . among the acceptable vehicles and solvents that may be employed are water , ringer &# 39 ; s solution and isotonic sodium chloride solution . in addition , sterile , fixed oils are conventionally employed as a solvent or suspending medium . for this purpose any bland fixed oil may be employed including synthetic mono - or diglycerides . in addition , fatty acids such as oleic acid find use in the preparation of injectables . the compounds of the present invention may also be administered in the form of suppositories for rectal administration of the drug . these compositions can be prepared by mixing the drug with a suitable non - irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug . such materials are cocoa butter and polyethylene glycols . for topical use , creams , ointments , jellies , solutions or suspensions , etc ., containing the compounds of the present invention are employed . compounds and compositions of the invention can be administered to mammals for veterinary use , such as for domestic animals , and clinical use in humans in a manner similar to other therapeutic agents . in general , the dosage required for therapeutic efficacy will vary according to the type of use and mode of administration , as well as the particularized requirements of individual hosts . ordinarily , dosages will range from about 0 . 001 to 1000 μg / kg , more usually 0 . 01 to 10 μg / kg , of the host body weight . alternatively , dosages within these ranges can be administered by constant infusion over an extended period of time , usually exceeding 24 hours , until the desired therapeutic benefits have been obtained . it will be understood , however , that the specific dose level and frequency of dosage for any particular patient may be varied and will depend upon a variety of factors including the activity of the specific compound employed , the metabolic stability and length of action of that compound , the age , body weight , general health , sex , diet , mode and time of administration , rate of excretion , drug combination , the severity of the particular condition , and the host undergoing therapy . the invention will now be described in greater detail by reference to the following non - limiting examples . hplc - purification of invention compounds was accomplished by rp - mplc on c18 - solid phase ( aldrich ) using a step gradient on kontes flex - columns ( 15 × 7 mm ). semipreparative hplc was performed on an isocratic hplc system with a waters pump 6000h on normal phase column si - dynamas - 60 å ( 250 × 5 mm ) or reversed phase column c18 - dynamax - 60 å , flow 2 ml / minute , with a differential refractomeric detector waters r401 . lc - ms — the lc - ms chromatography was performed on a hewlett - packard system series hp1100 with dad and msd1100 detection . the separation was accomplished on reversed phase c18 ( agilent hypersil ods 5 μm , column dimension 4 . 6 × 100 mm ), flow rate 0 . 7 ml / minute using a standard gradient : 10 % acetonitrile , 15 minutes ; 98 % acetonitrile ( burdick & amp ; jackson high purity solvents ). the ms - detection was in esi positive mode , capillary voltage 3500 ev , fragmentation voltage 70 ev , mass range m / z 100 - 1000 . the apci - mode was measured at a flow rate of 0 . 5 ml / minute , positive detection , capillary voltage 3000 ev , fragmentation voltage 70 ev . nmr - nmr spectra were measured on a varian 300 mhz gradient field spectrometer with inverse - mode for 1 h or 2d - nmr spectra . the 13c and dept spectra were measured on a varian 400 mhz , broad band instrument . thereference is set on the internal standard tetramethylsilane ( tms , 0 . 00 ppm ). ms - ei — low resolution ms - ei spectra were performed on a hewlett - packard mass spectrometer with magnetic sector field device , heating rate 20 ° c ./ minute up to 320 ° c ., direct injection inlet . ftms - maldi — high resolution ms data were obtained by maldi operating mode on an ionspec ultima ft mass spectrometer . ir — infrared spectra were measured on a perkin - elmer ft infrared spectrophotometer using nacl windows . isolation and characterization of “ salinsospora ” species , culture nos . cnb392 and cnb476 cnb392 and cnb476 possess signature nucleotides within their 16s rdna which separate these strains phylogenetically from all other members of the family micromonosporaceae ( see fig1 ) these signature nucleotides have been determined to be a definitive marker for members of this group which also have a physiological growth requirement of sodium . signature nucleotides were aligned to e . coli positions 27 - 1492 using all existing members of the micromonosporaceae in the ribosomal database project as of jan . 31 , 2001 . for the “ salinospora ” glade , 45 partially sequenced morphotypes displayed all the signature nucleotides from positions 207 - 468 . the seven “ salinospora ” isolates sequenced almost in their entirety ( see fig2 ) displayed all of the signatures in fig1 . the strains cnb392 and cnb476 form bright orange to black colonies on agar and lacks aerial mycelia . dark brown and bright orange diffusible pigments are produced depending upon cellular growth stage . spores blacken the colony surface and are borne on substrate mycelia . vegetative mycelia are finely branched and do not fragment . spores are produced singly or in clusters . neither sporangia nor spore motility has been observed for these strains . cnb392 and cnb476 have an obligate growth requirement for sodium and will not grow on typical media used for maintenance of other generic members of the micromonosporaceae . cnb392 and cnb476 have been found to grow optimally on solid media tcg or m1 at 30 ° c . tcg 3 grams tryptone m1 10 grams starch 5 grams casitone 4 grams yeast extract 4 grams glucose 2 grams peptone 18 grams agar ( optional ) 18 grams agar ( optional ) 1 liter filtered seawater 1 liter filtered seawater cnb392 and cnb476 are cultured in shaken a1bfe + c or cka - liquid media , 1 liter at 35 ° c . for 9 days . after 4 days 20 grams amberlite xad - 16 resin ( sigma , nonionic polymeric adsorbent ) is added . a1bfe + c 10 grams starch cka 5 grams starch 4 grams yeast 4 ml hydrosolubles extract ( 50 %) 2 grams peptone 2 grams menhaden meal 1 gram caco 3 2 grams kelp powder 5 ml kbr ( aqueous 2 grams chitosan solution , 20 1 liter filtered seawater grams / liter ) 5 ml fe 2 ( so 4 ) 3 × 4 h 2 o ( 8 grams / liter ) 1 liter filtered seawater the xad - 16 resin is filtered and the organic extract is eluted with 1 liter ethylacetate followed by 1 liter methanol . the filtrate is then extracted with ethylacetate ( 3 × 200 ml ). the crude extract from the xad adsorption is 105 mg . cytotoxicity on the human colon cancer cell hct - 116 assay is ic50 & lt ; 0 . 076 μg / ml . the crude extract was flash - chromatographed over c18 reversed phase ( rp ) using a step gradient ( fig5 ). the hct - 116 assay resulted in two active fractions , cnb392 - 5 and cnb392 - 6 . the combined active fractions ( 51 . 7 mg ), hct - 116 & lt ; 0 . 076 μg / ml ) were then chromatographed on an isocratic rp - hplc , using 85 % methanol at 2 ml / minute flow as eluent and using refractive index detection . the active fraction cnb392 - 5 / 6 ( 7 . 6 mg , hct - 116 & lt ; 0 . 076 μg / ml ) was purified on an isocratic normal phase hplc on silica gel with ethyl acetate : isooctane ( 9 : 1 ) at 2 ml / minute . salinosporamide a ( fig1 ) was isolated as a colorless , amorphous solid in 6 . 7 mg per 1 liter yield ( 6 . 4 %). tlc on silica gel ( dichloromethane : methanol 9 : 1 ) shows salinosporamide a at r f = 0 . 6 , no uv extinction or fluorescence at 256 nm , yellow with h 2 so 4 / ethanol , dark red - brown with godin reagent ( vanillin / h 2 so 4 / hclo 4 ). salinosporamide a is soluble in chcl 3 , methanol , and other polar solvents like dmso , acetone , acetonitrile , benzene , pyridine , n , n - dimethylormamide , and the like . 1 h nmr : ( d 5 - pyridine , 300 mhz ) 1 . 37 / 1 . 66 ( 2h , m , ch 2 ), 1 . 70 . 2 . 29 ( 2h , m , ch 2 ), 1 . 91 ( 2h , broad , ch 2 ), 2 . 07 ( 3h , s , ch 3 ), 2 . 32 / 2 . 48 ( 2h , ddd , 3 j = 7 . 0 hz , ch 2 ), 2 . 85 ( 1h , broad , m , ch ), 3 . 17 ( 1h , dd , 3 j = 10 hz , ch ), 4 . 01 / 4 . 13 ( 2h , m , ch 2 ), 4 . 25 ( 1h , d , 3 j = 9 . 0 hz , ch ), 4 . 98 ( 1h , broad , oh ), 5 . 88 , ( 1h , ddd , 3 j = 10 hz , ch ), 6 . 41 ( 1h , broad d , 3 j = 10 hz , ch ) 10 . 62 ( 1h , s , nh ). 13 c nmr / dept : ( d 5 - pyridine , 400 mhz ) 176 . 4 ( coor ), 169 . 0 ( conh ), 128 . 8 (═ ch ), 128 . 4 (═ ch ), 86 . 1 ( c q ), 80 . 2 ( c q ), 70 . 9 ( ch ), 46 . 2 ( ch ), 43 . 2 ( ch 2 ), 39 . 2 ( ch ), 29 . 0 ( ch 2 ), 26 . 5 ( ch 2 ), 25 . 3 ( ch 2 ), 21 . 7 ( ch 2 ), 20 . 0 ( ch 3 ) m / z : ( m + h ) + 314 , ( m + na ) + 336 ; fragments : ( m + h — co 2 ) + 292 , ( m + h — co 2 — h 2 o ) + 270 , 252 , 204 . cl pattern : ( m + h , 100 %) + 314 , ( m + h , 30 %) + 316 . m / z : ( m + h ) + 314 , fragments : ( m + h — co 2 — h 2 o ) + 270 , 252 , 232 , 216 , 160 . ei : m / z : 269 , 251 , 235 , 217 , 204 , 188 ( 100 %), 160 , 152 , 138 , 126 , 110 , 81 . ft - ir : ( cm − 1 ) 2920 , 2344 , s , 1819 m , 1702 s , 1255 , 1085 s , 1020 s , 797 s . salinosporamide a shows strong activity against human colon cancer cells with an ic 50 of 0 . 011 μg / ml ( see fig4 ). the screening on antibacterial or antifungal activity shows no significant activity , see table 1 . crystallization of a compound of structure i from ethyl acetate / iso - octane resulted in single , cubic crystals , which diffracted as a monoclinic system p2 ( 1 ). the unusual high unit - cell volume of 3009 å hosted four independent molecules in which different conformational positions were observed for the flexible chloroethyl substituent . the assignment of the absolute structure from the diffraction anisotropy of the chlorine substituent resolved the absolute stereochemistry of salinosporamide a as 2r , 3s , 4r , 5s , 6s ( fig1 and 17 ) with a flack parameter of 0 . 01 and an esd of 0 . 03 . although the invention has been described with reference to the above examples , it will be understood that modifications and variations are encompassed within the spirit and scope of the invention . accordingly , the invention is limited only by the following claims .

the present invention is based on the discovery that certain fermentation products of the marine actinomycete strains cnb392 and cnb476 are effective inhibitors of hyperproliferative mammalian cells . the cnb392 and cnb476 strains lie within the family micromonosporaceae , and the generic epithet salinospora has been proposed for this obligate marine group . the reaction products produced by this strain are classified as salinosporamides , and are particularly advantageous in treating neoplastic disorders due to their low molecular weight , low ic 50 values , high pharmaceutical potency , and selectivity for cancer cells over fungi .

the present invention contemplates use of 5 - asa ( mesalamine ) plus an antioxidant either together as separate molecular entities or when coupled chemically , to provide an enhanced therapeutic or prophylactic effect against inflammatory bowel diseases in mammals . the combination of mesalamine plus an antioxidant in a mammal model of colitis promotes healing and reduces inflammation to a significantly greater degree than either agent when used alone . the present invention provides an improved anti - inflammatory effectiveness of 5 - asa for use in the treatment of inflammatory bowel disease by the addition of an antioxidant . use of the presently claimed compositions result in a reduction in the need for additional anti - inflammatory agents , such as prednisone and imuran in the treatment of inflammatory disease ( thereby providing a significant cost benefit and reduction in drug - induced toxicity ). furthermore , the present invention can serve as a model for additional pharmacological approaches to other inflammatory and , possibly , neoplastic conditions within the gastrointestinal tract . as shown below experiments were performed to examine the ability of an antioxidant ( nac ) and 5 - asa , when administered separately or in combination to the distal colon , to affect mucosal healing and repair following colitis chemically - induced by tnbs or dss . male sprague dawley rats ( 200 - 250 grams ) were housed in cages containing contact bedding . rats were deprived of food for 24 hrs prior to the induction of colitis , but were allowed free access to water throughout the experiment . institutional approval for experimental protocols was provided by the research and animal care committees of the research services at the oklahoma city veterans administration medical center . following a 24 hr fast , rats were lightly anesthetized using isoflurane followed by insertion of a polyethylene catheter into the anus . the tip of the catheter was advanced 8 cm proximal from the anus and a single dose of tnbs ( sigma , st . louis , mo . ), dissolved in 50 % ethanol , was injected through the cannula ( 15 mg dissolved in a 0 . 6 ml volume of 50 % ethanol ) to induce colitis . following the administration of tnbs the animals were maintained in a head - down position for approximately 60 seconds to prevent leakage of the infusate . after 72 hrs , rats dosed initially with tnbs were randomly assigned to one of four treatment groups to receive daily intracolonic therapy in a manner similar to that used for tnbs administration . the groups were : a .) 5 - aminosalicyclic acid ( 5 - asa ) ( 100 mg / kg ) b .) n - acetylcysteine ( nac ) ( 40 mm ) plus 5 - asa ( 100 mg / kg ) c .) nac ( 40 mm ), or d .) saline . an additional group of rats ( group e ) served as control and received saline per rectum on day 0 and at subsequent intervals that corresponded to treatment protocols . rats were treated for either 5 or 8 days and then sacrificed by cervical dislocation . fig1 illustrates the design of these experiments . colitis was induced by oral administration of a 4 % solution of dss ( dextran sulfate sodium ) in drinking water for 4 days . experimental groups were randomized to include group 1 control , group 2 - 5 received dss in drinking water and groups 3 - 5 were further randomized to receive daily intracolonic treatment with 5 - asa ( 100 mg / kg ) plus nac ( 40 mm ), group 3 ; 5 - asa alone , group 4 ; and nac constituted group 5 . animals received treatment for 4 days . macroscopic indices of colonic injury were scored and tabulated . histological features that were examined included epithelial damage and mucosal ulceration . additional measures included determination of serum c - reactive protein ( crp ) levels and cytokine gene expression in colonic tissues performed by ribonuclease protection assay ( rpa ). upon sacrifice , the distal 8 cm of the colon was removed , opened by a longitudinal incision and rinsed with phosphate buffered saline to remove fecal material . macroscopic assessment of colitis was carried out by an independent observer who was unaware of the treatment groups . the criterial and scale of grading are listed in table 1 . inflammation was present if the mucosa was erythematous . ulceration of the mucosa was defined as a distinct break or interruption of the mucosa . once macroscopic damage was assessed full thickness colonic tissue samples were taken from the inflamed areas and either processed for histology or snap frozen in liquid nitrogen for subsequent cytokine measurement and determination of myeloperoxidase ( mpo ) activity . colonic tissue samples taken from the initial segment were immersed in 10 % phosphate buffered formalin and subsequently embedded in paraffin . sections of 3 micron thickness were cut and stained with hematoxylin and eosin . the slides were then evaluated by a pathologist for epithelial damage , architectural changes , mononuclear infiltration , polymorphonuclear leukocyte ( pmn ) infiltration and ulceration . the individual microscopic features of colitis were graded according to criteria noted in table 2 . in addition to scoring individual features of colitis , an aggregate score of colitis was tabulated by adding together individual scores , thus , providing a global assessment of colitis . myeloperoxidase activity was used as an indirect measure of the severity of colonic inflammation by pmns . whole thickness tissues taken following macroscopic assessment were weighed ( 100 mg ) and immediately snap frozen in liquid nitrogen for storage at − 80 . degree . c . the tissues were then removed from storage at − 80 ° c . and allowed to thaw on ice . once thawed , 1 ml of hexadecyltrimethylammonium bromide ( htab ) ( sigma , st . louis , mo .) containing 50 mm kh 2 po 4 ( sigma , st . louis , mo .) and 0 . 1m na 2 hpo 4 ( sigma , st . louis , mo .) was added per 100 mg tissue for homogenization . homogenates then underwent a series of four freeze / thaw cycles before finally being centrifuged at 12 , 000 × g for 10 minutes at 4 ° c . the supernatant was collected for measurement of mpo activity . horseradish peroxidase ( sigma , st . louis , mo .) was used as a standard ; stock solution of 0 . 5 mg / ml . tetramethylbenzidine ( tmb ) ( sigma , st . louis , mo .) was used as the substrate for carrying out the reaction . at the time of assay 25 μl of standard and sample were added to appropriately labeled tubes . tmb was added at a volume of 250 μl to initiate the reaction and 0 . 1m h 2 so 4 ( 250 μl ) was added after 10 minutes to terminate the reaction . the absorbance changes were read at 450 nm and recorded . results were expressed as ng / ml / g of tissue . following the manufacturer &# 39 ; s protocol , 1 ml of tri reagent ( sigma , st . louis , mo .) is used per 100 mg of tissue . the frozen tissue is placed into the tri reagent and immediately homogenized using a tissue macerator . the homogenates are transferred to microcentrifuge tubes and 0 . 2 ml of chloroform is added per ml of tri reagent used . the samples are shaken thoroughly and allowed to sit at room temp for 3 min . samples were then centrifuged at 12000 × g for 15 min at 40 ° c . after centrifugation , the upper aqueous phase was transferred to a new tube and 9 . 5 ml of 2 - propanol is added to precipitate the rna . after sitting at room temp for 10 min . the samples are centrifuged at 12000 × g for 15 min . the supernatants are discarded and the pellets are washed in 70 % ethanol followed by centrifugation at 12000 × g for 5 min . the ethanol was removed and the pellets are dissolved in 50 μl of depc treated water per sample . quantitation is performed by spectrophotometry at 260 nm and 280 nm . rpa analysis was performed with riboquant ™ multi - probe rnase protection assay system ( bd biosciences - pharmingen , san diego , calif .). to 20 μg of total rna in 8 μl of hybridization buffer and 2 μl of 32 p - labeled in vitro transcribed rna probes were added . the probes were transcribed from the multiprobe template set rck - 1 which as a panel of probes against il - 1a , il - 1b , il - 3 , il - 5 , il - 10 , il - 2 , tnfα , tnbβ , gapdh genes . probes were prepared according to the manufactures protocol . the rna samples containing labeled probes were incubated briefly at 90 ° c ., slowly cooled to 56 ° c . and hybridized overnight at that temperature . the samples were then slowly cooled to 30 ° c . thereafter , a mixture of rnases a and t1 was added to digest single - stranded rna leaving the double - stranded rna formed by the probes annealing to their cognate mrnas intact . following proteinase k digestion and phenol - chloroform extraction , the undigested double - stranded rna was precipitated with ammonium acetate and ethanol . after centrifugation at 14000 × g for 20 min ., the pellets are washed with 90 % ethanol and recentrifuged . the ethanol was removed and the pellets were dissolved in 5 μl of formamide loading dye . the double - stranded rnas were resolved on a 5 % polyacrylamide / urea gel run in tbe buffer . after electrophoresis , the gel was dried at 80 ° c . under vacuum and then exposed to a phosphorimager screen overnight . detection and quantitation of the resolved bands on the gel were performed on a storm phosphorimager ™ system ( amersham biosciences , piscataway , n . j .). all values in the figures and text are expressed as means ± standard error of the mean ( sem ). the statistical significance of any difference among groups was analyzed using student &# 39 ; s two - tailed t test for equal and unequal variance observations . p values of & lt ; 0 . 05 were considered to be statistically significant . the macroscopic injury observed at 8 days after rats received tnbs was 4 . 5 ± 0 . 5 . this value indicated that the distal 8 cm of colon contained more than 2 discrete ulcers in an area of inflammation ( fig2 ). in contrast , combination therapy with the nac plus 5 - asa combination acted synergistically to cause a significant reduction in macroscopic injury as reflected by an injury score of 2 . 6 ± 0 . 7 ; p & lt ; 0 . 05 . monotherapy with either nac or 5 - asa alone caused reduction in macroscopic injury ( 4 . 1 ± 0 . 6 and 3 . 0 ± 0 . 7 , respectively ) but these values did not achieve significance when compared to tnbs alone . visual evidence of colitis 11 days after tnbs was scored at 4 . 1 ± 0 . 2 ( fig3 ). animal treated with the nac plus 5 - asa combination for 8 days showed no evidence of mucosal inflammation on ulceration . the colonic mucosa in these animals appeared normal . monotherapy with either nac or 5 - asa alone showed mild inflammation and macroscopic injury scores were 1 . 2 ± 0 . 2 and 1 . 8 ± 0 . 2 respectively . the cumulative or aggregate microscopic colitis injury score for rats examined 8 days after tnbs was 9 . 4 ± 1 . 0 which represented moderately severe colitis ( fig4 ). individual therapy for 5 days with either 5 - asa or nac alone did not alter the aggregate injury score : 9 . 0 ± 0 . 9 and 9 . 0 ± 1 . 8 , respectively . therapy with the nac plus 5 - asa combination , however , caused significant reduction (− 44 %) in cumulative colitis injury and resulted in a score of 5 . 0 ± 1 . 2 , p & lt ; 0 . 02 . subset analysis of individual histological features , when compared to tnbs alone , indicated that therapy with the nac plus 5 - asa combination acted synergistically to significantly reduce the degree of epithelial damage , the extent of mucosal ulceration and the amount of mononuclear cell infiltration . eleven days post tnbs the indices of microscopic colitis were reduced slightly but not significantly from aggregate scores recorded at 8 days after tnbs : 7 . 0 ± 0 . 9 vs 9 . 5 ± 1 . 0 ; p & gt ; 0 . 1 . in contrast to the histological results after 5 days of treatment each of the intraluminal therapies for 8 days caused significant reduction in global measures of colitis ( fig5 ). 5 - asa and nac each when administered alone caused comparable decrease in aggregate injury of 46 and 53 %, respectively . combination therapy with the nac plus 5 - asa combination caused a 75 % reduction in cumulative colitis injury score : 1 . 8 ± 0 . 5 vs 7 . 0 ± 0 . 9 tnbs alone ; p & lt ; 0 . 001 . furthermore , comparisons between monotherapies with either 5 - asa or nac alone and combination therapy indicated that the nac plus 5 - asa combination acted synergistically to produce a significantly greater degree of healing than either nac or 5 - asa alone . mpo activity in colonic tissue 11 days after tnbs treatment was elevated greater than 25 fold above values derived from saline treated rats ( fig6 ). in contrast , the nac plus 5 - asa combination treatment for 8 days reduced mpo activity by 85 % to 100 . 9 ± 12 . 9 ng / g ( p & lt ; 0 . 004 vs tnbs 690 . 2 ± 101 . 5 ng / g ). mpo activity was also reduced by both nac and 5 - asa alone to levels that were 40 % and 32 % below tnbs values but these differences did not achieve statistical significance . cytokine gene expression in colonic tissues indicate that elevations in interleukins ( il 1a , il 1b , il - 4 and il - 6 ) and tnf α that were induced by tnbs after 11 days were inhibited significantly by 8 day treatment with the nac plus 5 - asa combination ( table 3 ). treatments with either nac or 5 - asa alone did not significantly suppress levels of cytokine expression induced by tnbs . results of the tnbs experiments of the present study indicate that intraluminal administration of the nac plus 5 - asa combination to the distal colon of rats with tnbs colitis act synergistically to cause a significant reduction in colonic inflammation and ulceration and acceleration of mucosal healing when compared to either agent used alone . furthermore , combination therapy with the nac plus 5 - asa combination caused significantly greater inhibition of myeloperoxidase activity and proinflammatory cytokine gene expression in colons of tnbs treated animals than either nac or 5 - asa alone . duration of treatment was a determinant in the effectiveness of antioxidant and anti - inflammatory agents on healing of chemically induced colitis . the nac plus 5 - asa combination was the only dosing regimen that resulted in significant improvement in both macroscopic and microscopic measures of colitis after a five day treatment . in contrast , eight day treatment with nac alone , 5 - asa alone or the nac plus 5 - asa combination resulted in significant improvement in histological features of colitis . treatment with the nac plus 5 - asa combination , however , caused greater improvement in mucosal injury , inflammation and epithelial regeneration than nac or 5 - asa alone . data derived from these studies indicate that intraluminal therapy with the antioxidant nac plus 5 - asa combination is superior to either agent alone in the treatment of tnbs colitis and that dual therapy has a synergistic effect in reducing inflammation and promoting mucosal repair . colitis induced by intracolonic instillation of tnbs manifests many of the histological and clinical features of colonic inflammatory bowel disease ( 11 ). this model of colonic ulceration and transmural inflammation of the mucosa ( 12 ) has been employed to study the pathogenesis of colonic inflammation and to investigate potential treatments of ibd . in this latter regard previous reports have shown that both 5 - asa and nac , when administered singly by intracolonic route to rats , ( 9 , 13 ) exerted an anti - inflammatory effect on tnbs colitis . furthermore , the antioxidant nac was observed to increase colonic glutathione stores which were associated with a reciprocal decrease in the extent of mucosal injury ( 9 ). in support of these observations and the role of ros generation in tnbs colitis loguercio et al reported that glutathione supplementation improved oxidative damage in tnbs colitis ( 14 ). antioxidants , such as nac , and 5 - asa possess the ability to scavenge oxygen free radicals , inhibit inducible no formation and to down regulate nuclear factor κb ( nf - κb ) activity ( 9 , 10 , 15 - 17 ). furthermore , antioxidants , such as phenyl n - tert - butylnitrone , and 5 - asa have been shown to inhibit cytokine production , including tumor necrosis factor tnfα , and to retard adhesion module expression and b - cell mediated antibody production ( 10 , 18 - 20 ) in experimented models of colitis . separately and together these agents can , thus , be envisioned to moderate immunocyte ( t cell ) mediated cytokine elaboration , neutrophil generation of ros and no , prostaglandin release and to facilitate an environment for unopposed cellular and growth factor - mediated tissue repair . these results show that treatment with the nac plus 5 - asa combination cause marked improvement in indices of colitis and , furthermore , demonstrate prominent features of epithelial repair , and mucosal architectural and glandular restoration . these data combined with the near normalization of mpo activity and marked reduction in cytokine ( ila , ilb , il6 ) expression indicate that therapy with the nac plus 5 - asa combination exerts a significantly greater anti - inflammatory and reparative effect in tnbs colitis than either 5 - asa or nac when used alone . results in the dss experiments demonstrated that dss under these experimental conditions produced mild to moderate colitis . the aggregate microscopic injury score for dss treated animals was 5 . 5 ± 2 . 0 ; maximal score 12 . monotherapy with either 5 - asa or nac alone caused slight reduction in aggregate scores to 4 . 1 ± 0 . 9 and 4 . 0 ± 1 . 1 , respectively . these values were not significantly different from dss alone . in contrast , combination therapy with the nac plus 5 - asa combination reduced global or aggregate histological injury score by 67 % to 1 . 8 ± 0 . 8 . furthermore , the nac plus 5 - asa combination caused significant improvement in epithelial damage when compared to dss alone . dss colitis was associated with elevated crp values of 7 . 0 ± 0 . 6 mg / ml . cpr levels were reduced substantially by concurrent therapy with nac plus 5 - asa and values were 1 . 0 ± 0 . 3 mg / ml . similar reductions in crp levels were observed with either 5 - asa or nac alone . dss treatment caused marked elevation in cytokine gene expression for il 1a and il 1b and these values represented a 6 . 8 and 12 . 1 fold increase , respectively , in gene expression of these cytokines when measured above control values . nac and 5 - asa , alone or in combination , substantially reduced dss - induced il1a and il1b gene expression by 55 - 90 % to levels that approximated control values . conclusions from these data indicate that intraluminal therapy with the nac plus 5 - asa combination caused significant amelioration of mucosal injury induced by dss . combination treatment with the nac plus 5 - asa combination results were associated with substantial reduction in serum crp levels and proinflammatory cytokine gene expression . current considerations of the pathogenesis of mucosal inflammation in ibd involve a number of steps from antigen presentation and processing by macrophages to amplification of t cell activation and differentiation and cytokine production ( 1 ). in addition , inflammatory cells , including granulocytes and mononuclear cells , are recruited to the mucosa in a highly coordinated fashion . once present in the inflamed mucosa , tissue injury is enhanced by neutrophil production of reactive oxygen species such as superoxide and an increase in the expression of the inducible isoform of no synthase ( inos ) ( 5 ). mucosal healing is thought to occur , in part , by reduction in injurious and proinflammatory substances and , also , by local liberation of growth factors which facilitate cellular restitution and repair ( 2 ). although several experimental approaches have been employed that suggest the importance of enhanced production of superoxide and nitric oxide in the pathogenesis of ibd , inconsistent results have this issue unresolved ( 5 ). the beneficial effect superoxide dismutase ( sod ) treatment in experimental models of colitis ( 21 - 23 ) has shown limited effect in humans with ibd ( 24 ). in addition , inos inhibition has shown variable results in experimental models of ibd ( 25 - 27 ). however , recent studies using gene - targeted mice suggest a dominant role of inos - derived no in a murine model of dextran sulfate sodium ( dss ) colitis ( 5 ). the antioxidants nac and phenyl n - tert butylnitrone have been shown to be effective in reducing the injurious consequences of tnbs colitis in rats ( 9 ) and dds colitis in mice ( 10 ), respectively . antioxidant therapy has also been demonstrated to suppress colonic inos activity and to decrease nf - κb dna - binding activity in experimental colitis ( 10 ) and man ( 16 ). in conclusion , our results showed that treatment of tnbs - induced colitis with the nac plus 5 - asa combination was superior to either 5 - asa or nac when used alone in reducing colonic inflammation and in promoting mucosal repair . in addition , combination therapy with the nac plus 5 - asa combination acted synergistically to result in , a significant reduction in mpo activity and proinflammatory cytokine gene expression . the present invention provides a method for the treatment of a mammal subject ( including humans ) afflicted with inflammatory diseases , and in particular , inflammatory bowel diseases . the present invention has several advantages over current therapies . as demonstrated herein , combination therapy with 5 - asa plus an antioxidant is synergistically superior to either agent alone in controlling mucosal inflammation . it is envisioned that such combination therapy would be capable of being delivered by oral route , employing existing coating technologies , to sites of inflammation . in addition , such combination therapy would be amenable to local therapy in the distal colon and rectum by enema or suppository . further description regarding deliver methods and dosing systems and protocols is discussed below . the combination of at least two active anti - inflammatory agents into a single delivery system as described herein provides greater clinical efficacy , development of a new pharmacochemical strategy for treating mucosal inflammatory conditions , and a reduction in the need for other potentially toxic and expensive anti - inflammatory agents . the chemical coupling of 5 - asa to an antioxidant substance can provide a further pharmacological approach to the treatment of mucosal inflammatory conditions such as ibd . the 5 - asa used in the composition of the present invention may be provided as the free acid , or as a pharmaceutically - acceptable salt or ester , for example as described in u . s . pat . no . 5 , 013 , 727 , the entirety of which is hereby expressly incorporated by reference herein . both 5 - asa and antioxidants such as nac and phenyl n - tert - butylnitrone have relatively low profiles for toxicity . 5 - asa may be associated with allergic reactions to the medications and should be avoided in patients with aspirin sensitivity . as with any nonsteroidal agent there exists potential for hepatic and renal toxicity . n - acetylcysteine or mucomyst ™ has had a wide experience in man for the treatment of acetaminophen hepatotoxicity . this agent has proven safe . other antioxidants contemplated for use herein in conjunction with 5 - asa include other aminosalicylates including 4 - aminosalicylic acid ( 4 - asa ), and n - acetyl - 5 - aminosalicylic acid ; other nonsteroidal anti - inflammatory drugs ( nsaids ) including those that inhibit cyclooxygenase i and / or ii , such as sulindac , celecoxib and refacoxib ; ascorbate ; vitamin c ; vitamin a ; vitamin e ; beta - carotene ; herbal agents such as milk thistle ; selenium ; iron in various ferric and ferrous formulations ; phospholipase a2 inhibitors , e . g ., carboxymethylcellulose - linked phosphatidylethanolamine ; superoxide dismutase mimetics , such as mn ( ii / iii ) tetrakis ( 1 - methyl - 4 - peridyl ) of nmtmpyp ; melatonin ; zolimid ; rebamipide ; and phenyl n - tert - butylnitrone ( pbn ); and combinations of any of the above . while is it contemplated that applications of the invention would be principally of treating mucosal inflammatory conditions associated with inflammatory bowel diseases , such as ulcerative colitis , crohn &# 39 ; s disease and behcet &# 39 ; s disease , the invention would also have application to other disorders of the gastrointestinal tract such as radiation and infective enteritis , ischemic injury to the gastrointestinal tract , infectious , caustic agent - induced gastrointestinal injury , hemorrhagic rectal ulcer , ileum pouchitis , ischemic enteritis and drug - induced colitis , mucous colitis , pseudomembranous enterocolitis , non - specific colonic ulcers , collagenous colitis , cathartic colon , ulcerative proctitis , idiopathic diffuse ulcerative non - granulomatous enteritis , non - steroidal anti - inflammatory drug - induced inflammations , celiac sprue and the like . furthermore , advances in our understanding of the pathogenesis of gastrointestinal malignancies suggest a role for prostaglandins generated by the cyclooxygenase enzymes to have a role in neoplasia . it has been well established that nonspecific and specific cyclooxygenase inhibitors can reduce the propensity to neoplasia and malignancy in experimental models and in human subjects . full appreciation of the role of prostaglandins in the cyclooxygenase system in the pathogenesis of gastrointestinal malignancy is not complete . however , it is conceivable that therapies which combine inhibition prostaglandin synthesis through the cyclooxygenase enzymes and antioxidant therapies may have a beneficial role in preventing gastrointestinal malignancies such as colorectal cancer . the term “ inflammation ” as used herein is meant to include reactions of both the specific and non - specific defense systems . a specific defense system reaction is a specific immune system reaction response to an antigen . examples of a specific defense system reaction include the antibody response to antigens such as rubella virus , and delayed - type hypersensitivity response mediated by t - cells ( as seen , for example , in individuals who test “ positive ” in the mantaux test ). a non - specific defense system reaction is an inflammatory response mediated by leukocytes incapable of immunological memory . such cells include granulocytes , macrophages , neutrophils , for example . examples of a non - specific defense system reaction include the immediate swelling at the site of a bee sting , the reddening and cellular infiltrate induced at the site of a burn and the collection of pmn leukocytes at sites of bacterial infection ( e . g ., pulmonary infiltrates in bacterial pneumonias , pus formation in abscesses ). although the invention is particularly suitable for cases of acute inflammation , it also has utility for chronic inflammation . types of inflammation that can be treated with the present invention include diffuse inflammation , traumatic inflammation , immunosuppression , toxic inflammation , specific inflammation , reactive inflammation , parenchymatous inflammation , obliterative inflammation , interstitial inflammation , croupous inflammation , and focal inflammation . a therapeutically effective amount of a composition of the present invention refers to an amount which is effective in controlling , treating or moderating the inflammatory response . the terms “ controlling ”, “ treating ” or “ moderating ” are intended to refer to all processes wherein there may be a slowing , interrupting , arresting , or stopping of the progression of the disease and does not necessarily indicate a total elimination of all disease symptoms . the term “ therapeutically effective amount ” is further meant to define an amount resulting in the improvement of any parameters or clinical symptoms characteristic of the inflammatory response . the actual dose will be different for the various specific molecules , and will vary with the patient &# 39 ; s overall condition , the seriousness of the symptoms , and counter indications . as used herein , the term “ subject ” or “ patient ” refers to a warm blooded animal such as a mammal which is afflicted with a particular inflammatory disease state . it is understood that guinea pigs , dogs , cats , rats , mice , horses , cattle , sheep , goats , pigs , llamas , and humans are among the examples of animals within the scope of the meaning of the term . a therapeutically effective amount of the compound used in the treatment described herein can be readily determined by the attending diagnostician , as one skilled in the art , by the use of conventional techniques and by observing results obtained under analogous circumstances . in determining the therapeutically effective dose , a number of factors are considered by the attending diagnostician , including , but not limited to : the species of mammal ; its size , age , and general health ; the specific disease or condition involved ; the degree of or involvement or the severity of the disease or condition ; the response of the individual subject ; the particular compound administered ; the mode of administration ; the bioavailability characteristic of the preparation administered ; the dose regimen selected ; the use of concomitant medication ; and other relevant circumstances . a therapeutically effective amount of the compositions of the present invention will generally contain sufficient active ingredient ( i . e ., the antioxidant and 5 - asa ) to deliver from about 0 . 1 μg / kg to about 6000 mg / kg ( weight of active ingredient / body weight of patient ). preferably , the composition will deliver at least 1 . 0 μg / kg to 1000 mg / kg , and more preferably at least 1 mg / kg to 100 mg / kg , although each dose of the composition may be more or less than these amounts . for example , the daily dose for an adult may be in the range of about 10 mg to 300 mg / kg , preferably in the range of about 20 mg to 300 mg / kg , especially in the range of 50 mg / kg to 200 mg / kg . also see u . s . pat . no . 5 , 013 , 727 which is incorporated by reference herein . practice of the method of the present invention comprises administering to a subject a therapeutically effective amount of the composition described herein , in any suitable systemic or local formulation , in an amount effective to deliver the dosages listed above . the dosage can be administered on a one - time basis , or ( for example ) from one to five times per day or once or twice per week , or continuously via a venous drip , depending on the desired therapeutic effect . as noted , preferred amounts and modes of administration are able to be determined by one skilled in the art . one skilled in the art of preparing formulations can readily select the proper form and mode of administration depending upon the particular characteristics of the compound selected , the disease state to be treated , the stage of the disease , and other relevant circumstances using formulation technology known in the art , described , for example , in remington &# 39 ; s pharmaceutical sciences , latest edition , mack publishing co . pharmaceutical compositions can be manufactured utilizing techniques known in the art . typically the therapeutically effective amount of the compound will be admixed with a pharmaceutically acceptable carrier . the compounds or compositions of the present invention may be administered by a variety of routes , for example , orally , intrarectally or parenterally ( i . e ., subcutaneously , intravenously , intramuscularly , intraperitoneally , or intratracheally ). for oral administration , the compounds can be formulated into solid or liquid preparations such as capsules , pills , tablets , lozenges , melts , powders , suspensions , or emulsions . solid unit dosage forms can be capsules of the ordinary gelatin type containing , for example , surfactants , lubricants and inert fillers such as lactose , sucrose , and cornstarch or they can be sustained release preparations . in another embodiment , the compounds of this invention can be tabletted with conventional tablet bases such as lactose , sucrose , and cornstarch in combination with binders , such as acacia , cornstarch , or gelatin , disintegrating agents such as potato starch or alginic acid , and a lubricant such as stearic acid or magnesium stearate . liquid preparations are prepared by dissolving the active ingredient in an aqueous or non - aqueous pharmaceutically acceptable solvent which may also contain suspending agents , sweetening agents , flavoring agents , and preservative agents as are known in the art . for parenteral administration , the compounds may be dissolved in a physiologically acceptable pharmaceutical carrier and administered as either a solution or a suspension . illustrative of suitable pharmaceutical carriers are water , saline , dextrose solutions , fructose solutions , ethanol , or oils of animal , vegetative , or synthetic origin . the pharmaceutical carrier may also contain preservatives , and buffers as are known in the art . the compounds of this invention can also be administered topically . this can be accomplished by simply preparing a solution of the compound to be administered , preferably using a solvent known to promote transdermal absorption such as ethanol or dimethyl sulfoxide ( dmso ) with or without other excipients . preferably topical administration will be accomplished using a patch either of the reservoir and porous membrane type or of a solid matrix variety . as noted above , the compositions can also include an appropriate carrier . for topical use , any of the conventional excipients may be added to formulate the active ingredients into a lotion , ointment , powder , cream , spray , or aerosol . for surgical implantation , the active ingredients may be combined with any of the well - known biodegradable and bioerodible carriers , such as polylactic acid and collagen formulations . such materials may be in the form of solid implants , sutures , sponges , wound dressings , and the like . in any event , for local use of the materials , the active ingredients usually be present in the carrier or excipient in a weight ratio of from about 1 : 1000 to 1 : 20 , 000 , but are not limited to ratios within this range . preparation of compositions for local use are detailed in remington &# 39 ; s pharmaceutical sciences , latest edition , ( mack publishing ). additional pharmaceutical methods may be employed to control the duration of action . increased half - life and controlled release preparations may be achieved through the use of polymers to conjugate , complex with , or absorb the composition described herein . the controlled delivery and / or increased half - life may be achieved by selecting appropriate macromolecules ( for example , polysaccharides , polyesters , polyamino acids , homopolymers polyvinyl pyrrolidone , ethylenevinylacetate , methylcellulose , or carboxymethylcellulose , and acrylamides such as n -( 2 - hydroxypropyl ) methacrylamide , and the appropriate concentration of macromolecules as well as the methods of incorporation , in order to control release . another possible method useful in controlling the duration of action by controlled release preparations and half - life is incorporation of the glycosulfopeptide molecule or its functional derivatives into particles of a polymeric material such as polyesters , polyamides , polyamino acids , hydrogels , poly ( lactic acid ), ethylene vinylacetate copolymers , copolymer micelles of , for example , peg and poly ( i - aspartamide ). alternatively , it is possible to entrap the compostions in microcapsules prepared , for example , by coacervation techniques or by interfacial polymerization ( for example , hydroxymethylcellulose or gelatine - microcapsules and poly -( methylmethacylate ) microcapsules , respectively ), in colloidal drug delivery systems ( for example , liposomes , albumin microspheres , microemulsions , nano - particles , and nanocapsules ), or in macroemulsions . such techniques are disclosed in the latest edition of remington &# 39 ; s pharmaceutical sciences . u . s . pat . no . 4 , 789 , 734 describe methods for encapsulating compositions in liposomes and is hereby expressly incorporated by reference herein . essentially , the material is dissolved in an aqueous solution , the appropriate phospholipids and lipids added , along with surfactants if required , and the material dialyzed or sonicated , as necessary . a review of known methods is by g . gregoriadis , chapter 14 . “ liposomes ”, drug carriers in biology and medicine , pp . 287 - 341 ( academic press , 1979 ). microspheres formed of polymers or proteins are well known to those skilled in the art , and can be tailored for passage through the gastrointestinal tract directly into the blood stream . alternatively , the agents can be incorporated and the microspheres , or composite of microspheres , implanted for slow release over a period of time , ranging from days to months . see , for example , u . s . pat . nos . 4 , 906 , 474 ; 4 , 925 , 673 ; and 3 , 625 , 214 which are expressly incorporated by reference herein . when the composition is to be used as an injectable material , it can be formulated into a conventional injectable carrier . suitable carriers include biocompatible and pharmaceutically acceptable phosphate buffered saline solutions , which are preferably isotonic . for reconstitution of a lyophilized product in accordance with this invention , one may employ a sterile diluent , which may contain materials generally recognized for approximating physiological conditions and / or as required by governmental regulation . in this respect , the sterile diluent may contain a buffering agent to obtain a physiologically acceptable ph , such as sodium chloride , saline , phosphate - buffered saline , and / or other substances which are physiologically acceptable and / or safe for use . in general , the material for intravenous injection in humans should conform to regulations established by the food and drug administration , which are available to those in the field . the pharmaceutical composition may also be in the form of an aqueous solution containing many of the same substances as described above for the reconstitution of a lyophilized product . the compounds can also be administered as a pharmaceutically acceptable acid - or base - addition salt , formed by reaction with inorganic acids such as hydrochloric acid , hydrobromic acid , perchloric acid , nitric acid , thiocyanic acid , sulfuric acid , and phosphoric acid , and organic acids such as formic acid , acetic acid , propionic acid , glycolic acid , lactic acid , pyruvic acid , oxalic acid , malonic acid , succinic acid , maleic acid , and fumaric acid , or by reaction with an inorganic base such as sodium hydroxide , ammonium hydroxide , potassium hydroxide , and organic bases such as mono -, di -, trialkyl and aryl amines and substituted ethanolamines . as mentioned above , the compounds of the invention may be incorporated into pharmaceutical preparations which may be used for therapeutic purposes . however , the term “ pharmaceutical preparation ” is intended in a broader sense herein to include preparations containing a 5 - asa / antioxidant composition in accordance with this invention , used not only for therapeutic purposes but also for reagent or diagnostic purposes as known in the art , or for tissue culture . the pharmaceutical preparation intended for therapeutic use should contain a “ pharmaceutically acceptable ” or “ therapeutically effective amount ” of the composition , i . e ., that amount necessary for preventative or curative health measures . if the pharmaceutical preparation is to be employed as a reagent or diagnostic , then it should contain reagent or diagnostic amounts of a 5 - asa / antioxidant combination . all references , patents and patent applications cited herein are hereby incorporated herein in their entirety by reference . the present invention is not to be limited in scope by the specific embodiments described herein , since such embodiments are intended as but single illustrations of one aspect of the invention and any functionally equivalent embodiments are within the scope of this invention . indeed , various modifications of the invention in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings . 1 . konstantions a , papadakis k a , targan s r . current theories on the causes of inflammatory bowel disease . gastroenterol clin north am 28 : 283 - 296 ; 1999 2 . sands b e . novel therapies for inflammatory bowel disease . gastroenterol clin north am 18 : 323 - 351 ; 1999 3 . sands b e . crohn &# 39 ; s disease . in : gastrointestinal and liver disease . eds . m feldman , l s friedman , m h sleisenger . saunders ; new york , 2002 pp 2005 - 2038 . 4 . grisham m b . oxidants and free radicals in inflammatory bowel disease . lancet b44 : 859 - 861 ; 1994 . 5 . krieglstein c f , cerwinka w h , laroux s , et al . regulation of murine intestinal inflammation by reactive metabolites of oxygen and nitrogen : divergent roles of superoxide and nitric oxide . j exp med 194 : 1207 - 1218 ; 2001 . 6 . rachmilewitz d , karmeli f , okon e , et al . experimental colitis is ameliorated by inhibition of nitric oxide synthase activity . gut 37 : 247 - 255 ; 1995 . 7 . yoshida y , iwai a , itoh m , et al . role of inducible nitric oxide synthase in dextran sulfate sodium - induced colitis . aliment pharmacol ther 14 : 26 - 32 ; 2000 . 8 . mccafferty d m , miampamba m , shiota e . role of inducible nitric oxide synthase in trinitrobenzene sulphonic acid induced colitis in mice . gut 45 : 199 - 209 ; 1999 . 9 . ardite e , sans m , panes ], et al . replenishment of glutathione levels improves mucosal function in experimental acute colitis . lab invest 80 : 735 - 744 ; 2000 . 10 . naito y . takagi t , ishikawa t , et al . . alpha . phenyl - n - tert - butylnitrone provides protection from dextran sulfate sodium - induced colitis in mice . antioxidants and redox signaling 4 : 195 - 206 ; 2002 . 11 . morris g p , beck p l , herridge m s , et al . hapten - induced model of chronic inflammation and ulceration in the rat colon . gastroenterology 1989 ; 96 : 795 - 803 . 12 . wallace j i . release of platelet - activating factor ( paf ) and accelerated healing induced by a paf antagonist in an animal model of chronic colitis . can i physiol pharmacol 1988 ; 66 : 422 - 425 . 13 . galvez j , gamido m , rodriguez - cabezas m e , et al . the intestinal anti - inflammatory activity of ur - 12746s on reactivated experimental colitis is mediated through down regulation of cytokine production . inflamm bowel dis 2003 ; 9 : 363 - 371 . 14 . loguercio c , d &# 39 ; argenio g , delle cave m , et al . glutathione supplementation improves oxidative damage in experimental colitis . dig liver dis 2003 ; 35 : 635 - 641 . 15 . grisham m b . a radical approach to treating inflammation . trends in pharmacological sci 2000 ; 21 : 119 - 120 . 16 . bantel h , berg c , vieth m , et al . mesalamine inhibits activation of transcription factor nf - kb in inflamed mucosa of patients with ulcerative colitis . am j gastroenterol 2000 ; 95 : 3452 - 3457 . 17 . macdermott r p . progress in understanding the mechanism of action of 5 - aminosalicylicacid . am j gastroenterol 2000 ; 95 : 3343 - 3345 . 18 . imai f , suzuki t , ishibashi t , et al . effect of sulfasalazine on b cell . clin exp rheumalol 1991 ; 9 : 256 - 264 . 19 . barrera p , boerbooms a m , van der putte l b a , et al . effects of antirheumatic agents on cytokines . sewin arthritis rheum 1996 ; 25 : 234 - 253 . 20 . green field s m , hambhin a s , shakor z s , et al . inhibition of leukocyte adhesion molecule upregulation by tumor necrosis factor . alpha . : a novel mechanism of action of sulfasalazine . gut 1993 ; 34 : 252 - 256 . 21 . keshavarzian , a , morgan g , sedghi s , gordon j h , and doria m . 1990 . role of reactive oxygen metabolites in experimental colitis . gut 31 : 786 - 790 . 22 . miller m j , mcneill h , mullane k m , caravella s j , and clark d a . 1988 . sod prevents damage and attenuates eicosanoid release in a rabbit model of necrotizing enterocolitis . am j physiol 255 : g556 - g565 . 23 . clark d a , formabaio d m , mcneill h , mullane k m , carvella s j and miller m j . 1988 . contribution of oxygen - derived free radicals to experimental necrotizing enterocolitis . am j physiol . 130 : 537 - 542 . 24 . rachmilewitz d , karmeli f , okon e , et al . experimental colitis is ameliorated by inhibition of nitric oxide synthase activity . gut 1995 ; 37 : 247 - 255 . 25 . yoshida y , iwai a , itoh k , tanaka m , kato s , hokari r , miyahara t , koyama h , miura s , and kobayashi m . 2000 . role of inducible nitric oxide synthase in dextran sulfate sodium - induced colitis . ailment pharmacol ther 14 : 26 - 32 . 26 . mccafferty d m , miampamba m , shiota e , sharkey k a , and kubes p . 1999 . role of inducible nitric oxide synthase in trinitrobenzene sulphonic acid induced colitis in mice . gut 45 : 864 - 873 . 27 . zingarelli b , szabo c , and salzman a l . 1999 . reduced oxidative and nitrosative damage in murine experimental colitis in the absence of inducible nitric oxide synthase . gut 45 : 199 - 209 . 28 . kubes p , and mccafferty d m . 2000 . nitric oxide and intestinal inflammation . am j med 109 : 150 - 158 . 29 . svarty n . salazopyrin . a new sulfanilamide preparation . acta med scand 1 10 : 577 1942 . 30 . dick a . p ., grayson m . j ., carpenter r . g ., et al . controlled trial of sulphosalazine in treatment of ulcerative colitis . gut 5 : 437 - 442 ; 1964 . 31 . misiewitcz j . j ., leonard - jones j . e ., connell a . m ., et al . controlled trial of sulphosalazine in maintenance therapy for ulcerative colitis . lancet 1 : 185 - 188 ; 1979 . 32 . das k . m ., chowdhury j . r ., zapp b ., et al . small bowel absorption of sulphasalazine and its hepatic metabolism in human beings , cats and rats . gastroenterology 77 : 28 - 284 ; 1979 . 33 . peppercorn m . a ., goldman p . the role of intestinal bacteria in the metabolism of salicylazosulphopyridine . j pharmacol exp ther 181 : 555 - 562 ; 1972 . 34 . schroder h ., campbell d . e . absorption , metabolism and excretion of salicylazosulphopyridine in man . clin pharmocol ther 13 : 539 - 552 ; 1972 . 35 . sharon p ., ligumsky m ., rachmilewitz d ., et al . role of prostaglandins in ulcerative colitis : enhanced production during active disease and inhibition by sulfasalazine . gastroenterology 75 : 638 - 640 ; 1975 . 36 . ronne i . a ., nielson o . h ., burhave k ., et al . sulfasalazine and its anti - inflammatory metabolite 5 - aminosalicylic acid : effect on arachidonic acid metabolism in human neutrophils and free radical scavenging . prostaglandins , thomboxane and leukotriene research 17 : 9 - 8 - 922 ; 1999 . 37 . papadakis k . a ., targan s . r . current theories on the causes of inflammatory bowel disease . gastroenterol clin north am 28 : 323 - 351 ; 1999 . 38 . stein r . b ., hanauer s . b . medical therapy for inflammatory bowel disease . gastroenterol clin north am 28 : 297 - 321 ; 1999 . 39 . hanauer s . b ., meyers s ., sachar d . b . the pharmacology of anti - inflammatory drugs in inflammatory bowel disease . in kirsner j . b ., shorter r . g . ( eds ) inflammatory bowel disease , ed 4 . baltimore , williams & amp ; wilkins pp . 643 - 663 , 1995 . 40 . emerit j . s ., pelletier s ., likforman j ., et al . phase ii trial of copper zinc superoxide dismutase ( cu , zn , sod ) in the treatment of crohn &# 39 ; s disease . free radic res common 12 : 563 - 596 ; 1991 . 41 . barnes p . j ., karin m . nuclear factor - kb : a pivotal transcription factor in chronic inflammatory disease . n engl j med 336 : 1066 - 1071 ; 1997 .

inflammatory bowel diseases are represented by two idiopathic disorders , which include ulcerative colitis and crohn &# 39 ; s disease . ulcerative colitis is restricted to the colon and involves uncertain and inflammation of the lining of the large intestine . crohn &# 39 ; s disease , on the other hand , can involve the mucosa of the small and / or large intestine and may involve deeper layers of the bowel wall . the present invention is a combination of 5 - aminosalicylic acid and one or more antioxidants for treating such inflammatory bowel diseases .

as used herein , tight junction antagonists prevent , inhibit or reduce the opening of tight junctions , for example , the opening of tight junctions induced by a tight junction agonist . a tight junction antagonist may bind to the receptor that mediates tight junction agonist induced opening of tight junctions . for example , a tight junction antagonist may bind to the zot receptor and prevent , inhibit , reduce or reverse the tight junction opening triggered by the tight junction agonist zot . as used herein a subject is any animal , e . g ., mammal , upon which methods of the invention may be practiced and / or to which materials of the present invention may be administered . subjects include , but are not limited to , humans . any antagonist of tight junction opening may be used in the practice of the present invention . for example , antagonists of the invention may comprise peptide antagonists . examples of peptide antagonists include , but are not limited to , peptides that comprise an amino acid sequence selected from the group consisting of examples of peptide antagonists include , but are not limited to , peptides that consist of an amino acid sequence selected from the group consisting of seq id nos : 1 - 24 . examples of peptide antagonists of tight junctions can be found in u . s . pat . nos . 6 , 458 , 925 , 6 , 670 , 448 , 6 , 936 , 689 , and 7 , 189 , 696 the entire contents of which are specifically incorporate herein by reference . when the antagonist is a peptide , any length of peptide may be used . generally , the size of the peptide antagonist will range from about 6 to about 100 , from about 6 to about 90 , from about 6 to about 80 , from about 6 to about 70 , from about 6 to about 60 , from about 6 to about 50 , from about 6 to about 40 , from about 6 to about 30 , from about 6 to about 25 , from about 6 to about 20 , from about 6 to about 15 , from about 6 to about 14 , from about 6 to about 13 , from about 6 to about 12 , from about 6 to about 11 , from about 6 to about 10 , from about 6 to about 9 , or from about 6 to about 8 amino acids in length . peptide antagonists of the invention may be from about 8 to about 100 , from about 8 to about 90 , from about 8 to about 80 , from about 8 to about 70 , from about 8 to about 60 , from about 8 to about 50 , from about 8 to about 40 , from about 8 to about 30 , from about 8 to about 25 , from about 8 to about 20 , from about 8 to about 15 , from about 8 to about 14 , from about 8 to about 13 , from about 8 to about 12 , from about 8 to about 11 , or from about 8 to about 10 amino acids in length . peptide antagonists of the invention may be from about 10 to about 100 , from about 10 to about 90 , from about 10 to about 80 , from about 10 to about 70 , from about 10 to about 60 , from about 10 to about 50 , from about 10 to about 40 , from about 10 to about 30 , from about 10 to about 25 , from about 10 to about 20 , from about 10 to about 15 , from about 10 to about 14 , from about 10 to about 13 , or from about 10 to about 12 amino acids in length . peptide antagonists of the invention may be from about 12 to about 100 , from about 12 to about 90 , from about 12 to about 80 , from about 12 to about 70 , from about 12 to about 60 , from about 12 to about 50 , from about 12 to about 40 , from about 12 to about 30 , from about 12 to about 25 , from about 12 to about 20 , from about 12 to about 15 , or from about 12 to about 14 amino acids in length . peptide antagonists of the invention may be from about 15 to about 100 , from about 15 to about 90 , from about 15 to about 80 , from about 15 to about 70 , from about 15 to about 60 , from about 15 to about 50 , from about 15 to about 40 , from about 15 to about 30 , from about 15 to about 25 , from about 15 to about 20 , from about 19 to about 15 , from about 15 to about 18 , or from about 17 to about 15 amino acids in length . the peptide antagonists can be chemically synthesized and purified using well - known techniques , such as described in high performance liquid chromatography of peptides and proteins : separation analysis and conformation , eds . mant et al ., c . r . c . press ( 1991 ), and a peptide synthesizer , such as symphony ( protein technologies , inc ); or by using recombinant dna techniques , i . e ., where the nucleotide sequence encoding the peptide is inserted in an appropriate expression vector , e . g ., an e . coli or yeast expression vector , expressed in the respective host cell , and purified therefrom using well - known techniques . typically , compositions , such as pharmaceutical compositions , comprising a tight junction antagonist ( e . g ., peptide tight junction antagonist ) comprise a pharmaceutically effective amount of the antagonist . the pharmaceutically effective amount of antagonist ( e . g ., peptide tight junction antagonist ) employed in any given composition may vary according to factors such as the disease state , age , sex , and weight of the individual . dosage regimens may be adjusted to provide the optimum therapeutic response . for example , a single bolus may be administered , several divided doses may be administered over time or the dose may be proportionally reduced or increased as indicated by the exigencies of the therapeutic situation . generally , the amount of antagonist used for preventing , ameliorating and / or treating a disease in a subject will be in the range of about 1 . 0 μg to 1 g , preferably about 1 mg to about 1000 mg , or from about 10 mg to about 100 mg , or from about 10 mg to about 50 mg , or from about 10 mg to about 25 mg of antagonist per dose . compositions of the invention may comprise one or more tight junction antagonists at a level of from about 0 . 1 wt % to about 20 wt %, from about 0 . 1 wt % to about 18 wt %, from about 0 . 1 wt % to about 16 wt %, from about 0 . 1 wt % to about 14 wt %, from about 0 . 1 wt % to about 12 wt %, from about 0 . 1 wt % to about 10 wt %, from about 0 . 1 wt % to about 8 wt %, from about 0 . 1 wt % to about 6 wt %, from about 0 . 1 wt % to about 4 wt %, from about 0 . 1 wt % to about 2 wt %, from about 0 . 1 wt % to about 1 wt %, from about 0 . 1 wt % to about 0 . 9 wt %, from about 0 . 1 wt % to about 0 . 8 wt %, from about 0 . 1 wt % to about 0 . 7 wt %, from about 0 . 1 wt % to about 0 . 6 wt %, from about 0 . 1 wt % to about 0 . 5 wt %, from about 0 . 1 wt % to about 0 . 4 wt %, from about 0 . 1 wt % to about 0 . 3 wt %, or from about 0 . 1 wt % to about 0 . 2 wt % of the total weight of the composition . compositions of the invention may comprise one or more tight junction antagonists at a level of about 0 . 1 wt %, about 0 . 2 wt %, about 0 . 3 wt %, about 0 . 4 wt %, about 0 . 5 wt %, about 0 . 6 wt %, about 0 . 7 wt %, about 0 . 8 wt %, or about 0 . 9 wt % based on the total weight of the composition . compositions of the invention may comprise one or more tight junction antagonists at a level of from about 1 wt % to about 20 wt %, from about 1 wt % to about 18 wt %, from about 1 wt % to about 16 wt %, from about 1 wt % to about 14 wt %, from about 1 wt % to about 12 wt %, from about 1 wt % to about 10 wt %, from about 1 wt % to about 9 wt %, from about 1 wt % to about 8 wt %, from about 1 wt % to about 7 wt %, from about 1 wt % to about 6 wt %, from about 1 wt % to about 5 wt %, from about 1 wt % to about 4 wt %, from about 1 wt % to about 3 wt %, or from about 1 wt % to about 2 wt % of the total weight of the composition . compositions of the invention may comprise one or more tight junction effectors at a level of about 1 wt %, about 2 wt %, about 3 wt %, about 4 wt %, about 5 wt %, about 6 wt %, about 7 wt %, about 8 wt %, or about 9 wt % based on the total weight of the composition . compositions of the invention may be formulated for pulmonary delivery ( e . g ., may be pulmonary dosage forms ). typically such compositions may be provided as pharmaceutical aerosols , e . g ., solution aerosols or powder aerosols . those of skill in the art are aware of many different methods and devices for the formation of pharmaceutical aerosols , for example , those disclosed by sciarra and sciarra , aerosols , in remington : the science and practice of pharmacy , 20th ed ., chapter 50 , gennaro et al . eds ., lippincott , williams and wilkins publishing co ., ( 2000 ). in one embodiment , the dosage forms are in the form of a powder aerosol ( i . e , comprise particles ). these are particularly suitable for use in inhalation delivery systems . powders may comprise particles of any size suitable for administration to the lung . powder formulations may optionally contain at least one particulate pharmaceutically acceptable carrier known to those of skill in the art . examples of suitable pharmaceutical carriers include , but are not limited to , saccharides , including monosaccharides , disaccharides , polysaccharides and sugar alcohols such as arabinose , glucose , fructose , ribose , mannose , sucrose , trehalose , lactose , maltose , starches , dextran , mannitol or sorbitol . in one embodiment , a powder formulation may comprise lactose as a carrier . powder formulations may be contained in any container known to those in the art . containers may be capsules of , for example , gelatin or plastic , or in blisters ( e . g . of aluminum or plastic ), for use in a dry powder inhalation device . in some embodiments , the total weight of the formulation in the container may be from about 5 mg to about 50 mg . in other embodiments , powder formulations may be contained in a reservoir in a multi - dose dry powder inhalation device adapted to deliver a suitable amount per actuation . powder formulations typically comprise small particles . suitable particles can be prepared using any means known in the art , for example , by grinding in an airjet mill , ball mill or vibrator mill , sieving , microprecipitation , spray - drying , lyophilisation or controlled crystallisation . typically , particles will be about 10 microns or less in diameter . particles for use in the compositions of the invention may have a diameter of from about 0 . 1 microns to about 10 microns , from about 0 . 1 microns to about 9 microns , from about 0 . 1 microns to about 8 microns , from about 0 . 1 microns to about 7 microns , from about 0 . 1 microns to about 6 microns , from about 0 . 1 microns to about 5 microns , from about 0 . 1 microns to about 4 microns , from about 0 . 1 microns to about 3 microns , from about 0 . 1 microns to about 2 microns , from about 0 . 1 microns to about 1 micron , from about 0 . 1 microns to about 0 . 5 microns , from about 1 micron to about 10 microns , from about 1 micron to about 9 microns , from about 1 micron to about 8 microns , from about 1 micron to about &# 39 ; 7 microns , from about 1 micron to about 6 microns , from about 1 micron to about 5 microns , from about 1 micron to about 4 microns , from about 1 micron to about 3 microns , from about 1 micron to about 2 microns , from about 2 microns to about 10 microns , from about 2 microns to about 9 microns , from about 2 microns to about 8 microns , from about 2 microns to about 7 microns , from about 2 microns to about 6 microns , from about 2 microns to about 5 microns , from about 2 microns to about 4 microns , or from about 2 microns to about 3 microns . in some embodiments , particles for use in the invention may be about 1 micron , about 2 microns , about 3 microns , about 4 microns , about 5 microns , about 6 microns , about 7 microns , about 8 microns , about 9 microns , or about 10 microns in diameter . in one embodiment , the dosage forms are in the form of a solution aerosol ( i . e ., comprise droplets ). typically , droplets will be about 10 microns or less in diameter . droplets for use in the compositions of the invention may have a diameter of from about 0 . 1 microns to about 10 microns , from about 0 . 1 microns to about 9 microns , from about 0 . 1 microns to about 8 microns , from about 0 . 1 microns to about 7 microns , from about 0 . 1 microns to about 6 microns , from about 0 . 1 microns to about 5 microns , from about 0 . 1 microns to about 4 microns , from about 0 . 1 microns to about 3 microns , from about 0 . 1 microns to about 2 microns , from about 0 . 1 microns to about 1 micron , from about 0 . 1 microns to about 0 . 5 microns , from about 1 micron to about 10 microns , from about 1 micron to about 9 microns , from about 1 micron to about 8 microns , from about 1 micron to about 7 microns , from about 1 micron to about 6 microns , from about 1 micron to about 5 microns , from about 1 micron to about 4 microns , from about 1 micron to about 3 microns , from about 1 micron to about 2 microns , from about 2 microns to about 10 microns , from about 2 microns to about 9 microns , from about 2 microns to about 8 microns , from about 2 microns to about 7 microns , from about 2 microns to about 6 microns , from about 2 microns to about 5 microns , from about 2 microns to about 4 microns , or from about 2 microns to about 3 microns . in some embodiments , particles and / or droplets for use in the invention may be about 1 micron , about 2 microns , about 3 microns , about 4 microns , about 5 microns , about 6 microns , about 7 microns , about 8 microns , about 9 microns , or about 10 microns in diameter . in addition to a tight junction antagonist , compositions of the invention may further comprise one or more additional therapeutic agents , particularly therapeutic agents conventionally used for treating lung conditions such as ali and ards . such additional therapeutic agents include , but are not limited to , steroids and other anti - inflammatory compounds . suitable therapeutic agents may include one or more of aminosalicylates , corticosteroids , immunomodulators , antibiotics , cytokines , chemokines and biologic therapies . compositions of the invention may also comprise one or more pharmaceutically acceptable excipients . suitable excipients include , but are not limited to , buffers , buffer salts , bulking agents , salts , surface active agents , acids , bases , sugars , and binders . the compositions of the invention can be used for preventing , slowing the onset of , ameliorating and / or treating ali and ards . typically , compositions may be administered one or more times each day in an amount suitable to prevent , reduce the likelihood of an attack of , or reduce the severity of an attack of the underlying disease condition . in some embodiments , compositions of the invention may be given repeatedly over a protracted period , i . e ., may be chronically administered . typically , compositions may be administered one or more times each day in an amount suitable to treat ali and ards . in some embodiments , compositions of the invention may be used to treat acute symptoms of ali and ards . typically , embodiments of this type will require administration of the compositions of the invention to a subject undergoing an acute episode in an amount suitable to reduce the severity of the symptoms . one or more administration may be used . a composition according to the present invention may be pre - mixed prior to administration , or can be formed in vivo when two or more components ( e . g ., a tight junction antagonist and an additional therapeutic agent ) are administered within 24 hours of each other . when administered separately , the components may be administered in either order ( e . g . tight junction antagonist first followed by additional therapeutic agent or additional therapeutic agent first followed by tight junction antagonist ). the components can be administered within a time span of about 12 hours , about 8 hours , about 4 hours , about 2 hours , about 1 hour , about 0 . 5 hour , about 0 . 25 hour , about 0 . 1 hour , about 1 minute , about 0 . 5 minute , or about 0 . 1 minute . administration of the compositions described above , e . g ., compositions comprising one or more tight junction antagonists and optionally comprising one or more additional therapeutic agents , may be by inhalation . for example , one or more tight junction antagonists and one or more additional therapeutic agents or a mixture thereof ; may be in inhalable form . an example of an inhalable form is an atomizable composition such as an aerosol comprising the tight junction antagonist , either alone or in combination with one or more additional therapeutic agents , in solution or dispersion in a propellant , or a nebulizable composition comprising a solution or dispersion of the active ingredient in an aqueous , organic or aqueous / organic medium . for example , the inhalable form of the compositions of the invention may be an aerosol comprising a mixture of one or more tight junction antagonists and one or more additional therapeutic agents in solution or dispersion in a propellant , or a combination of an aerosol containing one or more tight junction antagonists in solution or dispersion in a propellant with an aerosol containing one or more therapeutic agents in solution or dispersion in a propellant . in another example , the inhalable form of the compositions of the invention my be a nebulizable composition comprising a dispersion of one or more tight junction antagonists and one or more additional therapeutic agents in an aqueous , organic or aqueous / organic medium , or a combination of a dispersion of one or more tight junction antagonists with a dispersion of one or more additional therapeutic agents in such a medium . compositions of the invention may be administered in conjunction with one or more additional therapeutic agents , particularly therapeutic agents used conventionally for treating lung conditions such as ali and ards . such additional therapeutic agents include , but are not limited to , steroids and other anti - inflammatory compounds . suitable therapeutic agents may include one or more of aminosalicylates , corticosteroids , immunomodulators , antibiotics , cytokines , chemokines and biologic therapies . compositions of the invention and one or more additional therapeutic agents may be administered simultaneously , or alternatively compositions of the invention and one or more additional therapeutic agents may not be administered simultaneously . furthermore , compositions of the invention may be administered prior to administration of one or more additional therapeutic agents , or alternatively compositions of the invention may be administered subsequent to administration of one or more additional therapeutic agents . the following examples are provided for illustrative purposes only , and are in no way intended to limit the scope of the present invention . the igg immune complex model is a well established model of lung injury and is shown schematically in fig1 . briefly , a heterologous antibody mix containing antibodies to a known antigen are injected into an animal intravenously ( iv ) or intratracheally ( it ). the known antigen and a small amount of radiolabelled known antigen are injected into the animal iv . this results in immune complex ( ic ) formation between the antigen and the cognate antibodies in the heterologous antibody mix . the immune complex binds to binds to the fc gamma receptor ( fcγr ) and this initiates an inflammatory cascade and leads to injury . one of the results of the inflammatory cascade is an increase in lung permeability that increases with extent of injury . the increase in lung permeability is quantified by measuring the radiolabelled antigen present in lung versus blood where radiolabel in the lung versus blood increases with permeability . ( see johnson and ward , j . clin . investigation 54 : 349 - 357 , 1974 ). fig2 shows the results of an experiment where 4 - 5 mice per arm were treated as described above where the known antigen was bovine serum albumin ( bsa ) the heterologous antibody mix included antibody to bovine serum albumin ( anti - bsa ) and the treatment was carried out in the presence and absence of tight junction antagonist seq id no : 15 . fig2 shows a comparison of the change in lung permeability resulting from ic formation in response to the treatment described above in the presence and absence of tight junction antagonist as well as a comparison of the effects of iv versus it administration of tight junction antagonist . fig3 shows the results of varying the dose of seq id no : 15 administered with the anti - bsa antibodies delivered it . all publications , patents and patent applications mentioned in this specification are indicative of the level of skill of those skilled in the art to which this invention pertains , and are herein incorporated by reference to the same extent as if each individual publication , patent or patent application was specifically and individually indicated to be incorporated by reference .

the present application provides compositions and methods for treating acute lung injury and acute respiratory distress syndrome . the methods include administering one or more tight junction antagonists to the lung of a subject in need thereof .

unless otherwise defined herein , scientific and technical terms used in connection with the present invention have the meanings that are commonly understood by those of ordinary skill in the art . the phrase “ therapeutically effective ” is intended to qualify the amount of compound or pharmaceutical composition , or the combined amount of active ingredients in the case of combination therapy . this amount or combined amount will achieve the goal of treating the relevant condition . the term “ treatment ,” as used herein to describe the present invention and unless otherwise qualified , means administration of the compound , pharmaceutical composition or combination to effect preventative , palliative , supportive , restorative or curative treatment . the term treatment encompasses any objective or subjective improvement in a subject with respect to a relevant condition or disease . the term “ preventive treatment ,” as used herein to describe the present invention , means that the compound , pharmaceutical composition or combination is administered to a subject to inhibit or stop the relevant condition from occurring in a subject , particularly in a subject or member of a population that is significantly predisposed to the relevant condition . the term “ palliative treatment ,” as used herein to describe the present invention , means that the compound , pharmaceutical composition or combination is administered to a subject to remedy signs and / or symptoms of a condition , without necessarily modifying the progression of , or underlying etiology of , the relevant condition . the term “ supportive treatment ,” as used herein to describe the present invention , means that the compound , pharmaceutical composition or combination is administered to a subject as a part of a regimen of therapy , but that such therapy is not limited to administration of the compound , pharmaceutical composition or combination . unless otherwise expressly stated , supportive treatment may embrace preventive , palliative , restorative or curative treatment , particularly when the compounds or pharmaceutical compositions are combined with another component of supportive therapy . the term “ restorative treatment ,” as used herein to describe the present invention , means that the compound , pharmaceutical composition or combination is administered to a subject to modify the underlying progression or etiology of a condition . non - limiting examples include an increase in forced expiratory volume in one second ( fev 1 ) for lung disorders , decreased rate of a decline in lung function over time , inhibition of progressive nerve destruction , reduction of biomarkers associated and correlated with diseases or disorders , a reduction in relapses , improvement in quality of life , reduced time spent in hospital during an acute exacerbation event and the like . the term “ curative treatment ,” as used herein to describe the present invention , means that compound , pharmaceutical composition or combination is administered to a subject for the purpose of bringing the disease or disorder into complete remission , or that the disease or disorder is undetectable after such treatment . the term “ selective ”, when used to describe a functionally - defined receptor ligand or enzyme inhibitor means selective for the defined receptor or enzyme subtype as compared with other receptor or enzyme subtypes in the same family . for instance , a selective pde5 inhibitor is a compound which inhibits the pde5 enzyme subtype more potently than any other pde enzyme subtype . such selectivity is preferably at least 2 fold ( as measured using conventional binding assays ), more preferably at least 10 fold , most preferably at least 100 fold . the term “ alkyl ”, alone or in combination , means an acyclic , saturated hydrocarbon group of the formula c n h 2 + 1 which may be linear or branched . examples of such groups include methyl , ethyl , n - propyl , isopropyl , n - butyl , isobutyl , sec - butyl , tert - butyl , pentyl , iso - amyl and hexyl . unless otherwise specified , an alkyl group comprises from 1 to 6 carbon atoms . the carbon atom content of alkyl and various other hydrocarbon - containing moieties is indicated by a prefix designating a lower and upper number of carbon atoms in the moiety , that is , the prefix c i - c j indicates a moiety of the integer “ i ” to the integer “ j ” carbon atoms , inclusive . thus , for example , c 1 - c 6 alkyl refers to alkyl of one to six carbon atoms , inclusive . het 3 is a saturated or partially saturated ( i . e . non aromatic ) heterocycle and may be attached via a ring nitrogen atom ( when the heterocycle is attached to a carbon atom ) or a ring carbon atom ( in all cases ). equally , when substituted , the substituent may be located on a ring nitrogen atom ( if the substituent is joined through a carbon atom ) or a ring carbon atom ( in all cases ). specific examples include oxiranyl , aziridinyl , oxetanyl , azetidinyl , tetrahydrofuranyl , pyrrolidinyl , tetrahydropyranyl , piperidinyl , 1 , 4 - dioxanyl , morpholinyl , piperazinyl , azepanyl , oxepanyl , oxazepanyl and diazepinyl . het 3 may be fully saturated or partially unsaturated , i . e . may have one or more degrees of unsaturation but may not be fully aromatic . het 1 is an aromatic heterocycle and may be attached via a ring carbon atom ( in all cases ) or a ring nitrogen atom with an appropriate valency ( when the heterocycle is attached to a carbon atom ). equally , when substituted , the substituent may be located on a ring carbon atom ( in all cases ) or a ring nitrogen atom with an appropriate valency ( if the substituent is joined through a carbon atom ). specific examples include thienyl , furanyl , pyrrolyl , pyrazolyl , imidazoyl , oxazolyl , isoxazolyl , thiazolyl , isothiazolyl , triazolyl , oxadiazolyl , thiadiazolyl , tetrazolyl , pyridyl , pyridazinyl , pyrimidinyl and pyrazinyl . het 2 is an aromatic heterocycle and may be attached via a ring carbon atom ( in all cases ) or a ring nitrogen atom with an appropriate valency ( when the heterocycle is attached to a carbon atom ). equally , when substituted , the substituent may be located on a ring carbon atom ( in all cases ) or a ring nitrogen atom with an appropriate valency ( if the substituent is joined through a carbon atom ). het 2 is aromatic and is therefore necessarily a fused bicycle . specific examples include imidazo [ 2 , 1 - b ][ 1 , 3 ] thiazolyl , benzofuranyl , benzothienyl , indolyl , benzimidazolyl , indazolyl , benzotriazolyl , pyrrolo [ 2 , 3 - b ] pyridyl , pyrrolo [ 2 , 3 - c ] pyridyl , pyrrolo [ 3 , 2 - c ] pyridyl , pyrrolo [ 3 , 2 - b ] pyridyl , imidazo [ 4 , 5 - b ] pyridyl , imidazo [ 4 , 5 - c ] pyridyl , pyrazolo [ 4 , 3 - d ] pyridyl , pyrazolo [ 4 , 3 - c ] pyridyl , pyrazolo [ 3 , 4 - c ] pyridyl , pyrazolo [ 3 , 4 - b ] pyridyl , isoindolyl , indazolyl , purinyl , indolizinyl , imidazo [ 1 , 2 - a ] pyridyl , imidazo [ 1 , 5 - a ] pyridyl , pyrazolo [ 1 , 5 - a ] pyridyl , pyrrolo [ 1 , 2 - b ] pyridazinyl , imidazo [ 1 , 2 - c ] pyrimidinyl , quinolinyl , isoquinolinyl , cinnolinyl , quinazolinyl , quinoxalinyl , phthalazinyl , 1 , 6 - naphthyridinyl , 1 , 7 - naphthyridinyl , 1 , 8 - naphthyridinyl , 1 , 5 - naphthyridinyl , 2 , 6 - naphthyridinyl , 2 , 7 - naphthyridinyl , pyrido [ 3 , 2 - d ] pyrimidinyl , pyrido [ 4 , 3 - d ] pyrimidinyl , pyrido [ 3 , 4 - d ] pyrimidinyl , pyrido [ 2 , 3 - d ] pyrimidinyl , pyrido [ 2 , 3 - d ] pyrazinyl , pyrido [ 3 , 4 - b ] pyrazinyl , pyrimido [ 5 , 4 - d ] pyrimidinyl , pyrazino [ 2 , 3 - b ] pyrazinyl and pyrimido [ 4 , 5 - d ] pyrimidine . the term “ cycloalkyl ” means a means a monocyclic , saturated hydrocarbon group of the formula c n h 2n - 1 . examples include cyclopropyl , cyclobutyl , cyclopentyl , cyclohexyl , and cycloheptyl . unless otherwise specified , a cycloalkyl group comprises from 3 to 8 carbon atoms . the term “ oxo ” means a doubly bonded oxygen . the term “ alkoxy ” means a radical comprising an alkyl radical that is bonded to an oxygen atom , such as a methoxy radical . examples of such radicals include methoxy , ethoxy , propoxy , isopropoxy , butoxy and tert - butoxy . the term “ halo ” means , fluoro , chloro , bromo or iodo . as used herein , the terms “ co - administration ”, “ co - administered ” and “ in combination with ”, referring to a combination of a compound of formula ( i )-( ii ) and one or more other therapeutic agents , includes the following : simultaneous administration of such a combination of a compound of formula ( i )-( ii ) and a further therapeutic agent to a patient in need of treatment , when such components are formulated together into a single dosage form which releases said components at substantially the same time to said patient , substantially simultaneous administration of such a combination of a compound of formula ( i )-( ii ) and a further therapeutic agent to a patient in need of treatment , when such components are formulated apart from each other into separate dosage forms which are taken at substantially the same time by said patient , whereupon said components are released at substantially the same time to said patient , sequential administration of such a combination of a compound of formula ( i )-( ii ) and a further therapeutic agent to a patient in need of treatment , when such components are formulated apart from each other into separate dosage forms which are taken at consecutive times by said patient with a significant time interval between each administration , whereupon said components are released at substantially different times to said patient ; and sequential administration of such a combination of a compound of formula ( i )-( ii ) and a further therapeutic agent to a patient in need of treatment , when such components are formulated together into a single dosage form which releases said components in a controlled manner . the term ‘ excipient ’ is used herein to describe any ingredient other than a compound of formula ( i )-( ii ). the choice of excipient will to a large extent depend on factors such as the particular mode of administration , the effect of the excipient on solubility and stability , and the nature of the dosage form . the term “ excipient ” encompasses diluent , carrier or adjuvant . one way of carrying out the invention is to administer a compound of formula ( i )-( ii ) in the form of a prodrug . thus , certain derivatives of a compound of formula ( i )-( ii ) which may have little or no pharmacological activity themselves can , when administered into or onto the body , be converted into a compound of formula ( i )-( ii ) having the desired activity , for example by hydrolytic cleavage , particularly hydrolytic cleavage promoted by an esterase or peptidase enzyme . such derivatives are referred to as ‘ prodrugs ’. further information on the use of prodrugs may be found in ‘ pro - drugs as novel delivery systems ’, vol . 14 , acs symposium series ( t . higuchi and w . stella ) and ‘ bioreversible carriers in drug design ’, pergamon press , 1987 ( ed . e . b . roche , american pharmaceutical association ). reference can also be made to nature reviews / drug discovery , 2008 , 7 , 355 and current opinion in drug discovery and development , 2007 , 10 , 550 . prodrugs in accordance with the invention can , for example , be produced by replacing appropriate functionalities present in the compounds of formula ( i )-( ii ) with certain moieties known to those skilled in the art as ‘ pro - moieties ’ as described , for example , in ‘ design of prodrugs ’ by h . bundgaard ( elsevier , 1985 ). thus , a prodrug in accordance with the invention is ( a ) an ester or amide derivative of a carboxylic acid in a compound of formula ( i )-( ii ); ( b ) an ester , carbonate , carbamate , phosphate or ether derivative of a hydroxyl group in a compound of formula ( i )-( ii ); ( c ) an amide , imine , carbamate or amine derivative of an amino group in a compound form formula ( i )-( ii ); ( d ) a thioester , thiocarbonate , thiocarbamate or sulphide derivatives of a thiol group in a compound of formula ( i )-( ii ); or ( e ) an oxime or imine derivative of a carbonyl group in a compound of formula ( i )-( ii ). some specific examples of prodrugs in accordance with the invention include : ( i ) where a compound of formula ( i )-( ii ) contains a carboxylic acid functionality (— cooh ), an ester thereof , such as a compound wherein the hydrogen of the carboxylic acid functionality of the compound of formula ( i )-( ii ) is replaced by c 1 - c 8 alkyl ( e . g . ethyl ) or ( c 1 - c 8 alkyl ) c (═ o ) och 2 — ( e . g . t buc (═ o ) och 2 —); ( ii ) where a compound of formula ( i )-( ii ) contains an alcohol functionality (— oh ), an ester thereof , such as a compound wherein the hydrogen of the alcohol functionality of the compound of formula ( i )-( ii ) is replaced by — co ( c 1 - c 8 alkyl ) ( e . g . methylcarbonyl ) or the alcohol is esterified with an amino acid ; ( iii ) where a compound of formula ( i )-( ii ) contains an alcohol functionality (— oh ), an ether thereof , such as a compound wherein the hydrogen of the alcohol functionality of the compound of formula ( i )-( ii ) is replaced by ( c 1 - c 8 alkyl ) c (═ o ) och 2 — or — ch 2 op (═ o )( oh ) 2 ; ( iv ) where a compound of formula ( i )-( ii ) contains an alcohol functionality (— oh ), a phosphate thereof , such as a compound wherein the hydrogen of the alcohol functionality of the compound of formula ( i )-( ii ) is replaced by — p (═ o )( oh ) 2 or — p (═ o )( ona ) 2 or — p (═ o )( o ) 2 ca 2 + ; ( v ) where a compound of formula ( i )-( ii ) contains a primary or secondary amino functionality (— nh 2 or — nhr where r # h ), an amide thereof , for example , a compound wherein , as the case may be , one or both hydrogens of the amino functionality of the compound of formula ( i )-( ii ) is / are replaced by ( c 1 - c 10 ) alkanoyl , — coch 2 nh 2 or the amino group is derivatised with an amino acid ; ( vi ) where a compound of formula ( i )-( ii ) contains a primary or secondary amino functionality (— nh 2 or — nhr where r ≠ h ), an amine thereof , for example , a compound wherein , as the case may be , one or both hydrogens of the amino functionality of the compound of formula ( i )-( ii ) is / are replaced by — ch 2 op (═ o )( oh ) 2 . certain compounds of formula ( i )-( ii ) may themselves act as prodrugs of other compounds of formula ( i )-( ii ). it is also possible for two compounds of formula ( i )-( ii ) to be joined together in the form of a prodrug . in certain circumstances , a prodrug of a compound of formula ( i )-( ii ) may be created by internally linking two functional groups in a compound of formula ( i )-( ii ), for instance by forming a lactone . references below to compounds of formula ( i )-( ii ) are taken to include the compounds themselves and prodrugs thereof . the invention includes such compounds of formula ( i )-( ii ) as well as pharmaceutically acceptable salts of such compounds and pharmaceutically acceptable solvates of said compounds and salts . pharmaceutically acceptable salts of the compounds of formula ( i )-( ii ) include acid addition and base salts . suitable acid addition salts are formed from acids which form non - toxic salts . examples include the acetate , adipate , aspartate , benzoate , besylate , bicarbonate / carbonate , bisulfate / sulfate , borate , camsylate , citrate , cyclamate , edisylate , esylate , formate , fumarate , gluceptate , gluconate , glucuronate , hexafluorophosphate , hibenzate , hydrochloride / chloride , hydrobromide / bromide , hydroiodide / iodide , isethionate , lactate , malate , maleate , malonate , mesylate , methylsulfate , naphthylate , 2 - napsylate , nicotinate , nitrate , orotate , oxalate , palmitate , pamoate , phosphate / hydrogen phosphate / dihydrogen phosphate , pyroglutamate , saccharate , stearate , succinate , tannate , tartrate , tosylate , trifluoroacetate , naphatlene - 1 , 5 - disulfonic acid and xinofoate salts . suitable base salts are formed from bases which form non - toxic salts . examples include the aluminium , arginine , benzathine , calcium , choline , diethylamine , diolamine , glycine , lysine , magnesium , meglumine , olamine , potassium , sodium , tromethamine and zinc salts . hemisalts of acids and bases may also be formed , for example , hemisulfate and hemicalcium salts . for a review on suitable salts , see handbook of pharmaceutical salts : properties , selection , and use by stahl and wermuth ( wiley - vch , 2002 ). pharmaceutically acceptable salts of compounds of formula ( i )-( ii ) may be prepared by one or more of three methods : ( i ) by reacting a compound of formula ( i )-( ii ) with the desired acid or base ; ( ii ) by removing an acid - or base - labile protecting group from a suitable precursor of a compound of formula ( i )-( ii ) or by ring - opening a suitable cyclic precursor , for example , a lactone or lactam , using the desired acid or base ; or ( iii ) by converting one salt of the compound of formula ( i )-( ii ) to another by reaction with an appropriate acid or base or by means of a suitable ion exchange column . all three reactions are typically carried out in solution . the resulting salt may precipitate out and be collected by filtration or may be recovered by evaporation of the solvent . the degree of ionisation in the resulting salt may vary from completely ionised to almost non - ionised . the compounds of formula ( i )-( ii ), and pharmaceutically acceptable salts thereof , may exist in unsolvated and solvated forms . the term ‘ solvate ’ is used herein to describe a molecular complex comprising the compound of formula ( i )-( ii ), or a pharmaceutically acceptable salt thereof , and one or more pharmaceutically acceptable solvent molecules , for example , ethanol . the term ‘ hydrate ’ may be employed when said solvent is water . a currently accepted classification system for organic hydrates is one that defines isolated site , channel , or metal - ion coordinated hydrates — see polymorphism in pharmaceutical solids by k . r . morris ( ed . h . g . brittain , marcel dekker , 1995 ). isolated site hydrates are ones in which the water molecules are isolated from direct contact with each other by intervening organic molecules . in channel hydrates , the water molecules lie in lattice channels where they are next to other water molecules . in metal - ion coordinated hydrates , the water molecules are bonded to the metal ion . when the solvent or water is tightly bound , the complex will have a well - defined stoichiometry independent of humidity . when , however , the solvent or water is weakly bound , as in channel solvates and hygroscopic compounds , the water / solvent content will be dependent on humidity and drying conditions . in such cases , non - stoichiometry will be the norm . also included within the scope of the invention are multi - component complexes ( other than salts and solvates ) wherein the drug and at least one other component are present in stoichiometric or non - stoichiometric amounts . complexes of this type include clathrates ( drug - host inclusion complexes ) and co - crystals . the latter are typically defined as crystalline complexes of neutral molecular constituents which are bound together through non - covalent interactions , but could also be a complex of a neutral molecule with a salt . co - crystals may be prepared by melt crystallization , by recrystallization from solvents , or by physically grinding the components together . cf . chem . commun ., 17 , 1889 - 1896 , by o . almarsson and m . j . zaworotko ( 2004 ). for a general review of multi - component complexes , see j . pharm . sci ., 64 ( 8 ), 1269 - 1288 , by haleblian ( 1975 ). the compounds of the invention may exist in a continuum of solid states ranging from fully amorphous to fully crystalline . the term ‘ amorphous ’ refers to a state in which the material lacks long range order at the molecular level and , depending upon temperature , may exhibit the physical properties of a solid or a liquid . typically such materials do not give distinctive x - ray diffraction patterns and , while exhibiting the properties of a solid , are more formally described as a liquid . upon heating , a change from solid to liquid properties occurs which is characterised by a change of state , typically second order (‘ glass transition ’). the term ‘ crystalline ’ refers to a solid phase in which the material has a regular ordered internal structure at the molecular level and gives a distinctive x - ray diffraction pattern with defined peaks . such materials when heated sufficiently will also exhibit the properties of a liquid , but the change from solid to liquid is characterised by a phase change , typically first order (‘ melting point ’). the compounds of formula ( i )-( ii ) may also exist in a mesomorphic state ( mesophase or liquid crystal ) when subjected to suitable conditions . the mesomorphic state is intermediate between the true crystalline state and the true liquid state ( either melt or solution ). mesomorphism arising as the result of a change in temperature is described as ‘ thermotropic ’ and that resulting from the addition of a second component , such as water or another solvent , is described as ‘ lyotropic ’. compounds that have the potential to form lyotropic mesophases are described as ‘ amphiphilic ’ and consist of molecules which possess an ionic ( such as — coo − na + , — coo − k + , or — so 3 − n + ) or non - ionic ( such as — n − n + ( ch 3 ) 3 ) polar head group . for more information , see crystals and the polarizina microscope by n . h . hartshome and a . stuart , 4 th edition ( edward arnold , 1970 ). hereinafter all references to compounds of formula ( i )-( ii ) include references to pharmaceutically acceptable salts , solvates , multi - component complexes and liquid crystals thereof and to solvates , multi - component complexes and liquid crystals of pharmaceutically acceptable salts thereof . the compounds of formula ( i )-( ii ) may exhibit polymorphism and / or one or more kinds of isomerism ( e . g . optical , geometric or tautomeric isomerism ). the compounds of formula ( i )-( ii ) may also be isotopically labelled . such variation is implicit to the compounds of formula ( i )-( ii ) defined as they are by reference to their structural features and therefore within the scope of the invention . compounds of formula ( i )-( ii ) containing one or more asymmetric carbon atoms can exist as two or more stereoisomers . where a compound of formula ( i )-( ii ) contains an alkenyl or alkenylene group , geometric cis / trans ( or z / e ) isomers are possible . where structural isomers are interconvertible via a low energy barrier , tautomeric isomerism (‘ tautomerism ’) can occur . this can take the form of proton tautomerism in compounds of formula ( i )-( ii ) containing , for example , an imino , keto , or oxime group , or so - called valence tautomerism in compounds which contain an aromatic moiety . it follows that a single compound may exhibit more than one type of isomerism . the pharmaceutically acceptable salts of compounds of formula ( i )-( ii ) may also contain a counterion which is optically active ( e . g . d - lactate or l - lysine ) or racemic ( e . g . dl - tartrate or dl - arginine ). cis / trans isomers may be separated by conventional techniques well known to those skilled in the art , for example , chromatography and fractional crystallization . conventional techniques for the preparation / isolation of individual enantiomers include chiral synthesis from a suitable optically pure precursor or resolution of the racemate ( or the racemate of a salt or derivative ) using , for example , chiral high pressure liquid chromatography ( hplc ). alternatively , the racemate ( or a racemic precursor ) may be reacted with a suitable optically active compound , for example , an alcohol , or , in the case where the compound of formula ( i )-( ii ) contains an acidic or basic moiety , a base or acid such as 1 - phenylethylamine or tartaric acid . the resulting diastereomeric mixture may be separated by chromatography and / or fractional crystallization and one or both of the diastereoisomers converted to the corresponding pure enantiomer ( s ) by means well known to a skilled person . chiral compounds of formula ( i )-( ii ) ( and chiral precursors thereof ) may be obtained in enantiomerically - enriched form using chromatography , typically hplc , on an asymmetric resin with a mobile phase consisting of a hydrocarbon , typically heptane or hexane , containing from 0 to 50 % by volume of isopropanol , typically from 2 % to 20 %, and from 0 to 5 % by volume of an alkylamine , typically 0 . 1 % diethylamine . concentration of the eluate affords the enriched mixture . chiral chromatography using sub - and supercritical fluids may be employed . methods for chiral chromatography useful in some embodiments of the present invention are known in the art ( see , for example , smith , roger m ., loughborough university , loughborough , uk ; chromatographic science series ( 1998 ), 75 ( supercritical fluid chromatography with packed columns ), pp . 223 - 249 and references cited therein ). in some relevant examples herein , columns were obtained from chiral technologies , inc , west chester , pa ., usa , a subsidiary of daicel ® chemical industries , ltd ., tokyo , japan . when any racemate crystallizes , crystals of two different types are possible . the first type is the racemic compound ( true racemate ) referred to above wherein one homogeneous form of crystal is produced containing both enantiomers in equimolar amounts . the second type is the racemic mixture or conglomerate wherein two forms of crystal are produced in equimolar amounts each comprising a single enantiomer . while both of the crystal forms present in a racemic mixture have identical physical properties , they may have different physical properties compared to the true racemate . racemic mixtures may be separated by conventional techniques known to those skilled in the art . see , for example , stereochemistry of organic compounds by e . l . eliel and s . h . wilen ( wiley , 1994 ). the present invention includes all pharmaceutically acceptable isotopically - labelled compounds of formula ( i )-( ii ) wherein one or more atoms are replaced by atoms having the same atomic number , but an atomic mass or mass number different from the atomic mass or mass number which predominates in nature . isotopically - labelled compounds of formula ( i )-( ii ) can generally be prepared by conventional techniques known to those skilled in the art or by processes analogous to those described in the accompanying examples and preparations using an appropriate isotopically - labelled reagent in place of the non - labelled reagent previously employed . in particular , hydrogen atoms may be replaced by deuterium atoms since such deuterated compounds are sometimes more resistant to metabolism . also included within the scope of the invention are active metabolites of compounds of formula ( i )-( ii ), that is , compounds formed in vivo upon administration of the drug , often by oxidation or dealkylation . some examples of metabolites in accordance with the invention include ( i ) where a compound of formula ( i )-( ii ) contains a methyl group , an hydroxymethyl derivative thereof (— ch 3 -& gt ;— ch 2 oh ): ( ii ) where a compound of formula ( i )-( ii ) contains an alkoxy group , an hydroxy derivative thereof (— or -& gt ;— oh ); ( iii ) where a compound of formula ( i )-( ii ) contains a tertiary amino group , a secondary amino derivative thereof (— nrr ′-& gt ;— nhr or — nhr ′); ( iv ) where a compound of formula ( i )-( ii ) contains a secondary amino group , a primary derivative thereof (— nhr -& gt ;— nh 2 ); ( v ) where a compound of formula ( i )-( ii ) contains a phenyl moiety , a phenol derivative thereof (- ph -& gt ;- phoh ); and ( vi ) where a compound of formula ( i )-( ii ) contains an amide group , a carboxylic acid derivative thereof (— conh 2 -& gt ; cooh ). for administration to human patients , the total daily dose of a compound of formula ( i )-( ii ) is typically in the range of 0 . 01 mg to 500 mg depending , of course , on the mode of administration . in another embodiment of the present invention , the total daily dose of a compound of formula ( i )-( ii ) is typically in the range of 0 . 1 mg to 300 mg . in yet another embodiment of the present invention , the total daily dose of a compound of formula ( i )-( ii ) is typically in the range of 1 mg to 30 mg . the total daily dose may be administered in single or divided doses and may , at the physician &# 39 ; s discretion , fall outside of the typical range given herein . these dosages are based on an average human subject having a weight of about 65 kg to 70 kg . the physician will readily be able to determine doses for subjects whose weight falls outside this range , such as infants and the elderly . in the case of dry powder inhalers and aerosols , the dosage unit is determined by means of a prefilled capsule , blister or pocket or by a system that utilises a gravimetrically fed dosing chamber . units in accordance with the invention are typically arranged to administer a metered dose or “ puff ” containing from 1 to 5000 μg of drug . the overall daily dose will typically be in the range 1 μg to 20 mg which may be administered in a single dose or , more usually , as divided doses throughout the day . a compound of formula ( i )-( ii ) can be administered per se , or in the form of a pharmaceutical composition , which , as active constituent contains an efficacious dose of at least one compound of the invention , in addition to customary pharmaceutically innocuous excipients and / or additives . pharmaceutical compositions suitable for the delivery of compounds of the present invention and methods for their preparation will be readily apparent to those skilled in the art . such compositions and methods for their preparation may be found , for example , in reminaton &# 39 ; s pharmaceutical sciences , 19th edition ( mack publishing company , 1995 ). compounds of formula ( i )-( ii ) may be administered orally . oral administration may involve swallowing , so that the compound enters the gastrointestinal tract , or buccal or sublingual administration may be employed by which the compound enters the blood stream directly from the mouth . formulations suitable for oral administration include solid formulations such as tablets , capsules containing particulates , liquids , or powders , lozenges ( including liquid - filled ), chews , multi - and nano - particulates , gels , solid solution , liposome , films , ovules , sprays and liquid formulations . liquid formulations include suspensions , solutions , syrups and elixirs . such formulations may be employed as fillers in soft or hard capsules and typically comprise a carrier , for example , water , ethanol , polyethylene glycol , propylene glycol , methylcellulose , or a suitable oil , and one or more emulsifying agents and / or suspending agents . liquid formulations may also be prepared by the reconstitution of a solid , for example , from a sachet . compounds of formula ( i )-( ii ) may also be used in fast - dissolving , fast - disintegrating dosage forms such as those described in expert opinion in therapeutic patents , 11 ( 6 ), 981 - 986 , by liang and chen ( 2001 ). for tablet dosage forms , depending on dose , the drug may make up from 1 weight % to 80 weight % of the dosage form , more typically from 5 weight % to 60 weight % of the dosage form . in addition to the drug , tablets generally contain a disintegrant . examples of disintegrants include sodium starch glycolate , sodium carboxymethyl cellulose , calcium carboxymethyl cellulose , croscarmellose sodium , crospovidone , polyvinylpyrrolidone , methyl cellulose , microcrystalline cellulose , lower alkyl - substituted hydroxypropyl cellulose , starch , pregelatinised starch and sodium alginate . generally , the disintegrant will comprise from 1 weight % to 25 weight %. in one embodiment of the present invention , the disintegrant will comprise from 5 weight % to 20 weight % of the dosage form . binders are generally used to impart cohesive qualities to a tablet formulation . suitable binders include microcrystalline cellulose , gelatin , sugars , polyethylene glycol , natural and synthetic gums , polyvinylpyrrolidone , pregelatinised starch , hydroxypropyl cellulose and hydroxypropyl methylcellulose . tablets may also contain diluents , such as lactose ( monohydrate , spray - dried monohydrate , anhydrous and the like ), mannitol , xylitol , dextrose , sucrose , sorbitol , microcrystalline cellulose , starch and dibasic calcium phosphate dihydrate . tablets may also optionally comprise surface active agents , such as sodium lauryl sulfate and polysorbate 80 , and glidants such as silicon dioxide and talc . when present , surface active agents may comprise from 0 . 2 weight % to 5 weight % of the tablet , and glidants may comprise from 0 . 2 weight % to 1 weight % of the tablet . tablets also generally contain lubricants such as magnesium stearate , calcium stearate , zinc stearate , sodium stearyl fumarate , and mixtures of magnesium stearate with sodium lauryl sulfate . lubricants generally comprise from 0 . 25 weight % to 10 weight %. in one embodiment of the present invention , lubricants comprise from 0 . 5 weight % to 3 weight % of the tablet . other possible ingredients include anti - oxidants , colorants , flavoring agents , preservatives and taste - masking agents . exemplary tablets contain up to about 80 % drug , from about 10 weight % to about 90 weight % binder , from about 0 weight % to about 85 weight % diluent , from about 2 weight % to about 10 weight % disintegrant , and from about 0 . 25 weight % to about 10 weight % lubricant . tablet blends may be compressed directly or by roller to form tablets . tablet blends or portions of blends may alternatively be wet -, dry -, or melt - granulated , melt congealed , or extruded before tabletting . the final formulation may comprise one or more layers and may be coated or uncoated ; it may even be encapsulated . formulations of tablets are discussed in pharmaceutical dosage forms : tablets , vol . 1 , by h . lieberman and l . lachman ( marcel dekker , new york , 1980 ). consumable oral films for human or veterinary use are typically pliable water - soluble or water - swellable thin film dosage forms which may be rapidly dissolving or mucoadhesive and typically comprise a compound of formula ( i )-( ii ), a film - forming polymer , a binder , a solvent , a humectant , a plasticiser , a stabiliser or emulsifier , a viscosity - modifying agent and a solvent . some components of the formulation may perform more than one function . the film - forming polymer may be selected from natural polysaccharides , proteins , or synthetic hydrocolloids and is typically present in the range 0 . 01 to 99 weight %, more typically in the range 30 to 80 weight %. other possible ingredients include anti - oxidants , colorants , flavorings and flavor enhancers , preservatives , salivary stimulating agents , cooling agents , co - solvents ( including oils ), emollients , bulking agents , anti - foaming agents , surfactants and taste - masking agents . films in accordance with the invention are typically prepared by evaporative drying of thin aqueous films coated onto a peelable backing support or paper . this may be done in a drying oven or tunnel , typically a combined coater dryer , or by freeze - drying or vacuuming . solid formulations for oral administration may be formulated to be immediate and / or modified release . modified release includes delayed , sustained , pulsed , controlled , targeted and programmed release . suitable modified release formulations for the purposes of the invention are described in u . s . pat . no . 6 , 106 , 864 . details of other suitable release technologies such as high energy dispersions and osmotic and coated particles are to be found in pharmaceutical technology on - line , 25 ( 2 ), 1 - 14 , by verma et al ( 2001 ). the use of chewing gum to achieve controlled release is described in wo - a - 00 / 35298 . compounds of formula ( i )-( ii ) may also be administered directly into the blood stream , into muscle , or into an internal organ . such parenteral administration includes intravenous , intraarterial , intraperitoneal , intrathecal , intraventricular , intraurethral , intrasternal , intracranial , intramuscular , intra - articular and subcutaneous administration . suitable devices for parenteral administration include needle ( including microneedle ) injectors , needle - free injectors and infusion techniques . compounds of the invention may also be administered topically to the skin or mucosa , that is , dermally or transdermally . the compounds of formula ( i )-( ii ) can also be administered intranasally or by inhalation , typically in the form of a dry powder ( either alone , as a mixture , for example , in a dry blend with lactose , or as a mixed component particle , for example , mixed with phospholipids , such as phosphatidylcholine ) from a dry powder inhaler , as an aerosol spray from a pressurised container , pump , spray , atomiser ( preferably an atomiser using electrohydrodynamics to produce a fine mist ), or nebuliser , with or without the use of a suitable propellant , such as 1 , 1 , 1 , 2 - tetrafluoroethane or 1 , 1 , 1 , 2 , 3 , 3 , 3 - heptafluoropropane , or as nasal drops . for intranasal use , the powder may comprise a bioadhesive agent , for example , chitosan or cyclodextrin . delivery by inhalation is the preferred route of administration for the compounds of the present invention . the pressurised container , pump , spray , atomizer , or nebuliser contains a solution or suspension of the compound of formula ( i )-( ii ) comprising , for example , ethanol , aqueous ethanol , or a suitable alternative agent for dispersing , solubilising , or extending release of the compound , a propellant as solvent and an optional surfactant , such as sorbitan trioleate , oleic acid , or an oligolactic acid . prior to use in a dry powder or suspension formulation , the drug product is micronised to a size suitable for delivery by inhalation ( typically less than 5 microns ). this may be achieved by any appropriate comminuting method , such as spiral jet milling , fluid bed jet milling , supercritical fluid processing to form nanoparticles , high pressure homogenisation , or spray drying . capsules ( made , for example , from gelatin or hydroxypropylmethylcellulose ), blisters and cartridges for use in an inhaler or insufflator may be formulated to contain a powder mix of the compound of the invention , a suitable powder base such as lactose or starch and a performance modifier such as i - leucine , mannitol , or magnesium stearate . the lactose may be anhydrous or in the form of the monohydrate , preferably the latter . other suitable excipients include dextran , glucose , maltose , sorbitol , xylitol , fructose , sucrose and trehalose . a suitable solution formulation for use in an atomiser using electrohydrodynamics to produce a fine mist may contain from 1 μg to 20 mg of the compound of the invention per actuation and the actuation volume may vary from 1 μl to 1001 μl . a typical formulation may comprise a compound of formula ( i )-( ii ), propylene glycol , sterile water , ethanol and sodium chloride . alternative solvents which may be used instead of propylene glycol include glycerol and polyethylene glycol . suitable flavors , such as menthol and levomenthol , or sweeteners , such as saccharin or saccharin sodium , may be added to those formulations of the invention intended for intranasal administration . formulations for intranasal administration may be formulated to be immediate and / or modified release using , for example , pgla . modified release includes delayed , sustained , pulsed , controlled , targeted and programmed release . compounds of formula ( i )-( ii ) may also be administered directly to the eye or ear , typically in the form of drops of a micronised suspension or solution in isotonic , ph - adjusted , sterile saline . compounds of formula ( i )-( ii ) may be combined with soluble macromolecular entities , such as cyclodextrin and suitable derivatives thereof or polyethylene glycol - containing polymers , in order to improve their solubility , dissolution rate , taste , bioavailability and / or stability when using any of the aforementioned modes of administration . drug - cyclodextrin complexes , for example , are found to be generally useful for most dosage forms and administration routes . both inclusion and non - inclusion complexes may be used . as an alternative to direct complexation with the drug , the cyclodextrin may be used as an auxiliary additive , i . e . as a carrier , diluent , or solubilizer . most commonly used for these purposes are alpha -, beta - and gamma - cyclodextrins , examples of which may be found in international patent publications wo91 / 11172 , wo94 / 02518 and wo98 / 55148 . inasmuch as it may desirable to administer a combination of active compounds , for example , for the purpose of treating a particular disease or condition , it is within the scope of the present invention that two or more pharmaceutical compositions , at least one of which contains a compound of formula ( i )-( ii ), may conveniently be combined in the form of a kit suitable for coadministration of the compositions . thus , a kit of the invention comprises two or more separate pharmaceutical compositions , at least one of which contains a compound of formula ( i )-( ii ), and means for separately retaining said compositions , such as a container , divided bottle , or divided foil packet . an example of such a kit is the familiar blister pack used for the packaging of tablets , capsules and the like . such a kit is particularly suitable for administering different dosage forms , for example , oral and parenteral , for administering separate compositions at different dosage intervals , or for titrating the separate compositions against one another . to assist compliance , the kit typically comprises directions for administration and may be provided with a so - called memory aid . the compounds of the invention may be prepared by any method known in the art for the preparation of compounds of analogous structure . in particular , the compounds of the invention can be prepared by the procedures described by reference to the schemes that follow , or by the specific methods described in the examples , or by processes similar to either . the skilled person will appreciate that the experimental conditions set forth in the schemes that follow are illustrative of suitable conditions for effecting the transformations shown , and that it may be necessary or desirable to vary the precise conditions employed for the preparation of compounds of formula ( i )-( ii ). it will be further appreciated that it may be necessary or desirable to carry out the transformations in a different order from that described in the schemes , or to modify one or more of the transformations , to provide the desired compound of the invention . in addition , the skilled person will appreciate that it may be necessary or desirable at any stage in the synthesis of compounds of the invention to protect one or more sensitive groups , so as to prevent undesirable side reactions . in particular , it may be necessary or desirable to protect amino or carboxylic acid groups . the protecting groups used in the preparation of the compounds of the invention may be used in conventional manner . see , for example , those described in ‘ greene &# 39 ; s protective groups in organic synthesis ’ by theodora w greene and peter g m wuts , third edition , ( john wiley and sons , 1999 ), in particular chapters 7 (“ protection for the amino group ”) and 5 (“ protection for the carboxyl group ”), incorporated herein by reference , which also describes methods for the removal of such groups . all of the derivatives of the formula ( i )-( ii ) can be prepared by the procedures described in the general methods presented below or by routine modifications thereof . the present invention also encompasses any one or more of these processes for preparing the derivatives of formula ( i )-( ii ), in addition to any novel intermediates used therein . the person skilled in the art will appreciate that the following reactions may be heated thermally or under microwave irradiation . according to a first process , compounds of formula ( i ) may be prepared from compounds of formula ( ix ) and ( viii ), as illustrated by scheme 1 . wherein x is — conh —; hal is cl , br or iodo ; m is boronic acid or boranate ester ; pg 1 is tert - butyl , 2 , 4 - dimethoxybenzyl ; pg 2 is silylethoxymethyl , tetrahydropyranyl ; pg 3 is silylethoxymethyl , benzyl , or methyl . it may be necessary or desirable to interchange the protecting groups in this scheme to provide the highest yielding transformations . compounds of formulae ( x ), ( ix ), ( viii ) and ( iv ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( i )-( ii ) may be prepared from compounds of formula ( ii ) according to process step ( vi ), a deprotection step mediated by either an organic acid , a lewis acid or hydrogenation , or a sequential combination of each required . preferred conditions comprise tfa and / or boron tribromide in a suitable organic solvent such as dcm or neat , at room or elevated temperatures and / or hydrogenation using a suitable catalyst such as 10 % pd / c in an organic solvent such as etoh at room temperature . wherein compounds of formula ( i )-( ii ) are racemic , chiral separation may be employed to afford the two enantiomers . wherein compounds of formula ( i ) include an r group that contains oxooxazolidine , this may be reacted with a suitable organic base to effect an open chain r group . preferred conditions comprise sodium hydroxide at from 0 ° c . to room temperature for 18 hours . compounds of formula ( ii ) may be prepared from compounds of formula ( iii ) according to process step ( v ), a carbonylation step in the presence of a suitable amine of formula ( x ) or ( xi ), a suitable palladium catalyst , and an organic base and a suitable solvent heated either in a sealed tube or under microwave irradiation . typical conditions comprise molybdenum hexacarbonyl with dbu and palladium acetate heated to 100 ° c . either thermally for 45 minutes or under microwave irradiation for 10 minutes in the presence of a compound of formula ( x ) or ( xi ), such as methylamine or 88 % ammonia in a suitable organic solvent such as thf . alternatively carbon monoxide gas ( typically at 1 - 100 atmospheres ) can be used in place of molybdenum hexacarbonyl in the carbonylation step . compounds of formula ( iii ) may be prepared from compounds of formula ( iv ) and ( v ) according to process step ( iv ), an n - oxide rearrangement step with compounds of formula ( iv ) and an organic base in a suitable organic solvent at elevated temperatures . preferred conditions comprise triethylamine in dmf at elevated temperatures of between 80 - 100 ° c . for 18 hours . compounds of formula ( v ) may be prepared from compounds of formula ( vi ) according to process step ( iii ), an oxidation reaction . preferred conditions comprise mcpba in dcm at 0 ° c . for 18 hours . compounds of formula ( vi ) may be prepared from compounds of formula ( vii ) according to process step ( ii ), an electrophilic halogenation reaction . typically , compounds ( vii ) have the pg 2 protecting group removed by methods known to those skilled in the art prior to electrophilic halogenation . preferred conditions comprise n - iodosuccinimide in dmf at from 0 ° c . to room temperature for 18 hours followed by subsequent reprotection with pg 2 . compounds of formula ( vii ) may be prepared from compounds of formula ( ix ) and ( viii ) according to process step ( i ), a suzuki cross - coupling reaction with compounds of formula ( v ). suzuki cross - coupling is conveniently effected in the presence of a suitable catalyst , e . g ., palladium or nickel and a base . typical conditions comprise a boronic acid or ester , a palladium catalyst with phosphine ligands in an organic solvent at elevated temperatures . preferred suzuki conditions comprise palladium acetate with phosphine ligand s - phos , and potassium phosphate in ethanol at 80 ° c . for 18 hours . according to a second process , compounds of formula ( i ) may be prepared from compounds of formula ( vi ) as illustrated by scheme 2 . wherein x is — conh —; hal is cl , br or i , pg 1 is tert - butyl , 2 , 4 - dimethoxybenzyl ; pg 2 is silylethoxymethyl , tetrahydropyranyl ; pg 3 is silylethoxymethyl , benzyl , or methyl . compounds of formulae ( xii ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( vi ) are described in scheme 1 . compounds of formula ( i ) may be prepared from compounds of formula ( xiii ) according to process steps ( vii ) and ( vi ), a nucleophilic aromatic substitution reaction with compounds of formula ( xii ) followed by a deprotection step . typical conditions comprise heating to 90 ° c . with compounds of formula ( xii ) in a suitable organic solvent with a suitable organic base , followed by deprotection as described in scheme 1 . preferred conditions comprise dipea in n - butanol at 90 ° c . for 18 hours or triethylamine in dmf at 80 - 100 ° c . for 6 hours followed by tfa in dcm followed by boron tribromide in dcm . alternatively compounds of formula ( i ) may be prepared from compounds of formula ( xiii ) and formula ( xii ) using a cross coupling reaction followed by deprotection if required . typical conditions comprise a suitable metal catalyst in the presence of an inorganic base with an organic ligand . preferred conditions comprise pd 2 ( dba ) 3 with binap and cesium carbonate in toluene at elevated temperatures of 80 - 140 ° c . either thermally or under microwave irradiation . compounds of formula ( xiii ) may be prepared from compounds of formula ( xiv ) according to process steps ( iii ) and ( viii ), an oxidation reaction followed by an n - oxide rearrangement - halogenation reaction . typical conditions comprise oxidation as described in scheme 1 process step ( iii ) followed by stirring the n - oxide in a suitable organic solvent at temperatures of 0 - 10 ° c . with electrophilic halogenating reagents . preferred conditions comprise mcpba in dcm followed by either pocl 3 or oxalyl chloride in dcm . compounds of formula ( xiv ) may be prepared from compounds of formula ( vi ) and either ( x ) or ( xi ) according to process step ( v ) as described in scheme 1 . according to a third process , compounds of formula ( i ) may be prepared from compounds of formula ( iii ) as illustrated by scheme 3 . wherein x is — conh —; hal is cl , br or i ; pg 1 is tert - butyl , 2 , 4 - dimethoxybenzyl ; pg 2 is silylethoxymethyl , tetrahydropyranyl ; pg 3 is silylethoxymethyl , benzyl , or methyl . compounds of formulae ( xi ) and ( xi ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( iii ) are described in scheme 1 . compounds of formula ( i ) may be prepared from compounds of formula ( ii ) according to process step ( vi ) as described in scheme 1 . compounds of formula ( ii ) may be prepared from compounds of formula ( xv ) according to process step ( ix ), an amide bond formation reaction with compounds of formula ( x ) or ( xi ) with activation of the carboxylic acid via a mixed anhydride or using a suitable base such as dipea and a suitable coupling agent such as hatu , bop . preferred conditions comprise isobutyl chloroformate in thf with nmm or bop or hatu in dmf with dipea as base . process step ( vi ) may be performed before process step ( ix ) to obtain compounds of formula ( i ) in scheme 3 . compounds of formula ( xv ) may be prepared from compounds of formula ( iii ) according to process step ( v ) as described in scheme 1 but in the absence of compounds of formula ( x ) and ( xi ) in a solvent such as methanol with water added if necessary . according to a fourth process , compounds of formula ( i ) may be prepared from compounds of formula ( xx ) and ( xiv ) as illustrated by scheme 4 . wherein x is — conh —; hal is cl , br or i ; pg 1 is tert - butyl , 2 , 4 - dimethoxybenzyl ; pg 2 is silylethoxymethyl , tetrahydropyranyl ; pg 3 is silylethoxymethyl , benzyl , or methyl ; pg 4 is carboxybenzyl . compounds of formulae ( xvii ), ( x ) and ( xi ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( xiv ) are described in scheme 2 . compounds of formula ( xxi ) are described in scheme 5 . compounds of formula ( i ) may be prepared from compounds of formula ( xvi ) according to process step ( vi ) as described in scheme 1 . compounds of formula ( xvi ) may be prepared from compounds of formula ( xvii ) and ( xviii ) according to process step ( ix ), a sulfonamide formation step . preferred conditions comprise reacting compounds of formula ( xvii ) with compounds of formula ( xviii ) in a suitable organic solvent such as thf at from 0 ° c . to room temperature for 18 hours . alternatively a base may be added to facilitate the reaction such as sodium hydride . compounds of formula ( xviii ) may be prepared from compounds of formula ( xix ) according to process step ( vi ) a deprotection reaction as described in scheme 1 . preferred conditions comprise palladium on carbon in ethanol at room temperature under hydrogenation at 30 psi for 1 hour . compounds of formula ( xix ) may be prepared from compounds of formula ( xiv ) according to process steps ( iii ) and ( iv ), an oxidation of compounds of formula ( xiv ) followed by a rearrangement step with compounds of formula ( xx ) as described in scheme 1 . according to a fifth process , compounds of formula ( iv ) may be prepared from compounds of formula ( xxiv ) as illustrated by scheme 5 . compounds of formulae ( xxiv ), ( xvii ) and ( xxvi ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( iv ) may be prepared from compounds of formula ( xxi ) according to process step ( xv ), a reaction to form a carbamate activating group in the presence of an inorganic base . preferred conditions comprise sodium carbonate in dcm with 4 - nitrophenylchloroformate . compounds of formula ( xxi ) may be prepared from compounds of formula ( xxii ) according to process step ( xiv ), a reduction step in the presence of a metal catalyst and an inorganic hydrogen donor or under an atmosphere of hydrogen . preferred conditions comprise nicl 2 . 6h 2 o with sodium borohydride and di - tert butyl dicarbonate in methanol followed by 4m hcl in dioxane or 10 % palladium on carbon in acetic acid or raney nickel in methanolic ammonia under an atmosphere of 40 psi of hydrogen at room temperature for 18 hours . compounds of formula ( xxii ) may be prepared from compounds of formula ( xxv ) according to process step ( xiii ), an alkylation reaction with compounds of formula ( xxvi ) in the presence of a quaternary ammonium salt . preferred conditions comprise benzyltriethylammonium chloride and 40 % aqueous sodium hydroxide solution in thf with compounds of formula ( xxvi ). compounds of formula ( xxii ) may also be prepared from compounds of formula ( xxiii ) according to process step ( xii ), an alkylation reaction in the presence of an inorganic base . preferred conditions comprise potassium carbonate in acetone with compounds of formula ( xxvi ) or mitsunobu conditions with compounds of formula ( xviii ) using dead in thf . compounds of formulae ( xxv ) and ( xxiii ) may be prepared from compounds of formula ( xxiv ) and ( xvii ) according to process step ( xi ) a sulfonamide formation reaction . preferred conditions comprise stirring in pyridine at from 0 ° c . to room temperature or in the presence of lihmds in thf . compounds of formula ( xxiii ) may also be prepared from sulfonamides reacting with halo - substituted heterocycles in the presence of a base such as cesium carbonate in acetonitrile . according to a sixth process , compounds of formula ( ii ) may be prepared from compounds of formula ( vii ) as illustrated by scheme 6 . compounds of formula ( vii ) may be prepared as described in scheme 1 . compounds of formulae ( xii ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( i ) may be prepared from compounds of formula ( xxix ) according to reaction step ( vi ), a deprotection step as described in scheme 1 . compounds of formula ( xxix ) may be prepared according to reaction step ( xvi ), an n - oxide rearrangement step effected by employment of a dehydrating agent such as pybrop with amines of formula ( xii ). preferred conditions comprise pybrop with dipea in a suitable organic solvent such as dcm at room temperature . alternatively the n - oxide rearrangement step may employ acetic anhydride to afford the hydroxy intermediate followed by interconversion to triflate . the triflate may then be converted to compounds of formula ( xxix ) by heating with amines of formula ( xii ). typical conditions comprise heating the n - oxide with triethylamine and acetic anhydride , followed by triflic anhydride with pyridine in dcm at room temperature , and finally heating with compounds of formula ( xii ) with triethylamine in dmf . compounds of formula ( xxviii ) may be prepared from compounds of formula ( vii ) according to process step ( iii ) as described in scheme 1 . according to a seventh process , compounds of formula ( i ) may be prepared from compounds of formula ( vii ) as illustrated by scheme 7 . compounds of formula ( iii ) may be prepared as described in scheme 1 . compounds of formulae ( xxx ) or ( xxxi ) are commercially available or may be synthesized by those skilled in the art according to the literature or preparations described herein . compounds of formula ( i )-( ii ) may be prepared from compounds of formula ( iii ) according to process steps ( xvii ) and ( vi ), a cross coupling reaction , such as a stille reaction or a buchwald reaction followed by a deprotection step if required . typical conditions for a stille cross coupling reaction comprise a suitable tin reagent in the presence of one or two metal catalysts in a suitable organic solvent at elevated temperatures with compounds of formula ( xxx ). preferred conditions comprise bis ( tributyltin ) and copper ( i ) iodide with tetrakis ( triphenylphosphine ) palladium in toluene at 100 ° c . typical conditions for a buchwald reaction comprise a copper catalyst and a suitable organic ligand in the presence of an inorganic base at elevated temperatures . preferred conditions comprise cuprous oxide and 4 . 7 - dimethoxy - 1 , 10 - phenanthroline with cesium carbonate and peg in dmso at 110 ° c . compounds of formula ( i ) may also be prepared from compounds of formula ( xxxv ) according to process step ( xviii ) a heterocyclic cyclization reaction , either directly from the nitrile or the carboxylic acid , or via an acyl hydrazone from the carboxylic acid . preferred conditions comprise heating with the required nitrile or hydrazone in butanol at elevated temperatures under microwave irradiation . compounds of formula ( xxxv ) may be prepared from compounds of formula ( iii ) according to process step ( v ) as described in scheme 3 to afford the carboxylic acid , or using zinc cyanide and tetrakis ( triphenylphosphine ) palladium in dmf at elevated temperatures under microwave irradiation to afford the nitrile . according to an eighth process , compounds of formula ( i )-( ii ) may be prepared from compounds of formula ( xxxiii ) as illustrated by scheme 8 . compounds of formula ( xxxiii ) are either commercially available or prepared as described herein . compounds of formula ( i )-( i ) may be prepared from compounds of formula ( xxxii ) and ( viii ) according to process steps ( i ) and ( vi ), a suzuki cross - coupling reaction followed by deprotection as described in scheme 1 . preferred conditions for the suzuki step comprise peppsi - ipr catalyst with potassium carbonate in toluene at elevated temperatures . compounds of formula ( xxxii ) may be prepared from compounds of formula ( xxxiii ) and ( xii ) according to process step ( vii ), a nucleophilc aromatic substitution reaction as described in scheme 2 . the skilled person will further appreciate that compounds of formula ( i )-( ii ) may be interconverted to other compounds of formula ( i )-( ii ) by functional group manipulation , or suitably protected compounds of formula ( i )-( ii ) may be interconverted to other suitably protected compounds of formula ( i )-( ii ) followed by a deprotection step to afford compounds of formula ( i )-( ii ) wherein r or r 0 contains a ketone or aldehyde functionality , these may be reduced using a suitable reducing agent such as sodium borohydride ; wherein r or r 0 contain an amine , these may be interconverted to a urea , an amide , a sulfonamide or a sulfamide followed by suitable deprotection as required . wherein compounds of formula ( xvi ) contain an r ″ group that has a leaving group such as halo , an alkylation may occur with amines such as morpholine . in the non - limiting examples and preparations that are set out later in the description , and in the aforementioned schemes , the following the abbreviations , definitions and analytical procedures may be referred to : 1 h and 19 f nuclear magnetic resonance ( nmr ) spectra were in all cases consistent with the proposed structures . characteristic chemical shifts ( δ ) are given in parts - per - million downfield from tetramethylsilane ( for 1 h - nmr ) and upfield from trichloro - fluoro - methane ( for 19 f nmr ) using conventional abbreviations for designation of major peaks : e . g . s , singlet ; d , doublet ; t , triplet ; q , quartet ; m , multiplet ; br , broad . the following abbreviations have been used for common solvents : cdcl 3 , deuterochloroform ; d 6 - dmso , deuterodimethylsulphoxide ; and cd 3 od , deuteromethanol . where appropriate , tautomers may be recorded within the nmr data ; and some exchangeable protons may not be visible . mass spectra , ms ( m / z ), were recorded using either electrospray ionisation ( esi ) or atmospheric pressure chemical ionisation ( apci ). where relevant and unless otherwise stated the m / z data provided are for isotopes 19 f , 35 cl , 79 br and 127 i . wherein preparative tlc or silica gel chromatography has been used , one skilled in the art may choose any combination of solvents to purify the desired compound . either iupac or acd labs naming packages have been used , and are interchangeably employed throughout the examples and preparations . where singleton compounds are purified by preparative hplc , these are two methods used , shown below : detectors : elsd ; polymer labs pl - els 2100 , uv ; waters 2487 detector at 225 and 255 nm column : gemini nx c18 , 5 μm 21 . 2 × 100 mm ; temperature : ambient ; detection : elsd - ms ; mobile phase a : 0 . 1 % formic acid in water ; mobile phase b : 0 . 1 % formic acid in acetonitrile ; gradient : initial 0 % b , 1 min — 5 % b ; 7 mins — 95 % b ; 9 mins — 95 % b ; 9 . 1 mins — 5 % b ; 10 mins — 5 % b ; flow rate : 18 ml / min ; injection volume : 1000 μl column : gemini nx c18 , 5 μm 21 . 2 × 100 mm ; temperature : ambient ; detection : elsd - ms ; mobile phase a : 0 . 1 % diethylamine in water ; mobile phase b : 0 . 1 % diethylamine in acetonitrile ; gradient : initial 0 % b , 1 min — 5 % b ; 7 mins — 95 % b ; 9 mins — 95 % b ; 9 . 1 mins — 5 % b ; 10 mins — 5 % b ; flow rate : 18 ml / min ; injection volume : 1000 μl column : gemini c18 50 × 4 . 6 mm , 3 micron ; 5 minutes run . gradient initial — 95 % a , 5 % b ; 3 mins — 95 % b ; hold to 4 mins then back to 5 % b at 4 . 1 - 5 mins . flow rate 1 . 5 ml / min acidic conditions : mobile phase a : 0 . 1 % formic acid in water . mobile phase b : 0 . 1 % formic acid in acetonitrile basic conditions : mobile phase a : 0 . 1 % ammonia in water ; mobile phase b : 0 . 1 % ammonia in acetonitrile . to a solution of n - ethyl - n -( 2 -{[( 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethyl - silyl ) ethoxy ] methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ] methyl } phenyl ) ethanesulfonamide ( preparation 62 , 170 mg , 0 . 18 mmol ) in 2m methylamine in thf ( 2 . 6 ml ) was added molybdenum hexacarbonyl ( 48 . 18 mg , 0 . 181 mmol ), dbu ( 82 . 77 ml , 0 . 544 mmol ) and palladium acetate ( 2 . 85 mg , 0 . 01 mmol ). the reaction was heated at 100 ° c . under microwave irradiation for 10 minutes . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography eluting with 30 % etoac in hexanes . the residue was dissolved in tfa ( 3 ml ) and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh , cooled in ice and treated with ethylene diamine . the reaction was stirred at room temperature for 2 hours before concentrating in vacuo . the residue was purified using silica gel column chromatography eluting with etoac to afford the title compound ( 60 mg , 58 % over 2 steps ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 90 ( t , 3h ), 1 . 15 ( t , 3h ), 2 . 85 ( d , 3h ), 3 . 21 ( m , 2h ), 3 . 44 - 3 . 63 ( m , 4h ), 4 . 72 - 4 . 74 ( m , 1h ), 4 . 86 - 4 . 88 ( m , 1h ), 6 . 69 ( s , 1h ), 6 . 92 ( d , 1h ), 7 . 19 ( d , 1h ), 7 . 31 ( m , 2h ), 7 . 40 ( m , 2h ), 8 . 85 ( m , 1h ), 9 . 77 ( m , 1h ), 10 . 07 ( s , 1h ), 13 . 71 ( s , 1h ). the following examples ( examples 2 - 25 ) were prepared according to the method described for example 1 using the appropriate pyrazolo - pyridine and purification method ( pm ) below if different from the method described : purification method a : silica gel column chromatography eluting with between 40 - 60 % etoac in hexanes . purification method b : silica gel column chromatography or preparative tlc eluting with 4 % meoh in dcm . purification method c : silica gel column chromatography followed by preparative tlc eluting both with up to 30 % meoh in dcm . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 06 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 97 ( d , 6h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 83 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 02 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 04 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 21 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 92 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 21 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 05 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 97 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 28 ( m , 2h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 80 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 88 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 64 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 90 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 89 ( t , 3h ), 3 . 03 ( s , 3h ), 3 . 17 ( d , 3h ), 3 . 31 ( m , 2h ), 4 . 70 ( br m , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 01 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 85 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 95 ( t , 3h ), the following examples ( examples 26 - 28 ) were prepared according to the method described for example 1 using ammonia in thf and the appropriate pyrazolo - pyridine and purification method ( pm ) below if different from the method described . purification method e : silica gel column chromatography or preparative tlc eluting with 4 % meoh in dcm . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 03 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 95 ( t , 3h ), to a solution of 6 -( 2 - ethyl - 5 - fluoro - 4 - hydroxyphenyl )- 4 -({ 4 - methoxy - 2 -[ methyl ( methyl - sulfonyl ) amino ] benzyl } amino )- n - methyl - 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( example 28 , 80 mg , 0 . 14 mmol ) in dcm ( 10 ml ) was added boron tribromide ( 0 . 09 ml , 1 mmol ) at 0 ° c . the reaction was stirred at room temperature for 2 hours before the addition of further boron tribromide ( 0 . 09 ml , 1 mmol ) and further stirring for 2 hours . the reaction was partitioned between dcm and saturated aqueous sodium bicarbonate solution , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative hplc to afford the title compound ( 32 mg , 41 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 94 ( t , 3h ), 2 . 59 ( m , 2h ), 2 . 81 ( d , 3h ), 3 . 02 ( s , 3h ), 3 . 07 ( s , 3h ), 4 . 69 ( br m , 2h ), 6 . 61 ( s , 1h ), 6 . 73 ( dd , 1h ), 6 . 79 - 6 . 83 ( m , 2h ), 7 . 05 ( d , 1h ), 7 . 23 ( d , 1h ), 8 . 76 ( m , 1h ), 9 . 49 ( m , 1h ), 9 . 60 ( br s , 1h ). ms m / z 543 [ m + h ] + the title compound was prepared according to the method described for example 29 using 6 -( 2 - ethyl - 5 - fluoro - 4 - hydroxyphenyl )- 4 -({ 4 - methoxy - 2 -[ methyl ( methylsulfonyl ) amino ]- benzyl } amino )- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( example 27 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 96 ( t , 3h ), 2 . 60 ( m , 2h ), 3 . 01 ( s , 3h ), 3 . 08 ( s , 3h ), 4 . 65 ( br m , 2h ), 6 . 61 ( s , 1h ), 6 . 74 ( dd , 1h ), 6 . 80 - 6 . 83 ( m , 2h ), 7 . 09 ( d , 1h ), 7 . 25 ( d , 1h ), 7 . 77 ( br s , 1h ), 8 . 13 ( br s , 1h ), 9 . 49 ( m , 1h ), 9 . 56 ( s , 1h ), 9 . 76 ( s , 1h ), 13 . 56 ( s , 1h ). ms m / z 529 [ m + h ] + to a solution of n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl )- ethoxy ] meth - oxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ]- pyridin - 4 - yl ]- amino } methyl )- 4 - methoxyphenyl ]- n - methylmethanesulfonamide ( preparation 83 , 250 mg , 0 . 28 mmol ) in 2m methylamine in thf ( 3 ml ) was added dbu ( 0 . 13 ml , 0 . 85 mmol ), palladium acetate ( 4 . 43 mg , 0 . 02 mmol ) and molybdenum hexacarbonyl ( 75 mg , 0 . 28 mmol ) and the reaction was heated to 100 ° c . for 10 minutes under microwave irradiation . the reaction was cooled , concentrated in vacuo and purified directly using silica gel column chromatography eluting with 45 % etoac in hexanes . the resulting oil was dissolved in dcm ( 15 ml ) and cooled to 0 ° c . bbr 3 ( 0 . 10 ml , 1 . 07 mmol ) was added and the reaction stirred at room temperature for 6 hours . the reaction was concentrated in vacuo and partitioned between saturated aqueous sodium bicarbonate solution and etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc eluting with 5 % meoh in dcm to afford the title compound as a white solid ( 43 mg , 27 % over two steps ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 88 ( t , 3h ), 2 . 84 ( d , 3h ), 2 . 98 ( s , 3h ), 3 . 05 ( s , 3h ), 3 . 31 ( m , 2h ), 4 . 61 ( m , 1h ), 4 . 86 ( m , 1h ), 6 . 66 ( m , 2h ), 6 . 78 ( m , 2h ), 7 . 05 ( d , 1h ), 7 . 26 ( d , 1h ), 8 . 80 ( m , 1h ), 9 . 51 ( s , 1h ), 9 . 59 ( m , 1h ), 9 . 74 ( s , 1h ), 13 . 62 ( s , 1h ). ms m / z 543 [ m + h ] + to a solution of n -( 2 -{[( 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ] methyl }- phenyl )- 3 - methoxy - n - methylbenzenesulfonamide ( preparation 106 , 330 mg , 0 . 37 mmol ) in 2m methylamine in thf ( 2 ml ) was added dbu ( 0 . 16 ml , 1 . 19 mmol ), palladium acetate ( 5 . 86 mg , 0 . 03 mmol ) and molybdenum hexacarbonyl ( 99 mg , 0 . 37 mmol ) and the reaction was heated to 100 ° c . for 10 minutes under microwave irradiation . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography eluting with 35 % etoac in hexanes . the resulting oil was treated with tfa ( 0 . 5 ml ) and the solution stirred at room temperature for 30 minutes before concentrating in vacuo . ethylene diamine ( 0 . 5 ml ) was added and the reaction stirred at room temperature for 15 minutes before pouring onto ice - water and extracting into 20 % ipa in dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 45 % etoac in hexanes . the residue was dissolved in dcm ( 10 ml ) and boron tribromide ( 0 . 18 ml , 1 . 89 mmol ) was added dropwise at 0 ° c . the reaction was stirred at room temperature for 2 hours followed by quenching with saturated aqueous sodium bicarbonate solution and extracting into 20 % ipa in dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography followed by preparative tlc eluting both with 57 % etoac in hexanes to afford the title compound as a yellow solid ( 25 mg , 10 % over 3 steps ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 85 ( d , 3h ), 3 . 01 ( s , 3h ), 3 . 59 - 3 . 65 ( m , 2h ), 4 . 71 ( m , 1h ), 4 . 95 ( m , 1h ), 6 . 63 ( d , 1h ), 6 . 71 ( s , 1h ), 6 . 96 ( d , 1h ), 7 . 02 ( m , 1h ), 7 . 09 ( m , 2h ), 7 . 14 - 7 . 29 ( m , 3h ), 7 . 41 ( m , 2h ), 8 . 86 ( m , 1h ), 9 . 78 ( m , 1h ), 10 . 10 ( s , 1h ), 10 . 15 ( s , 1h ), 13 . 71 ( s , 1h ). ms m / z 659 [ m + h ] + the following examples ( examples 33 - 37 ) were prepared according to the method described for example 32 using the appropriate pyrazolo - pyridine , and purification method ( pm ) below if different from the method described : purification method f : silica gel column chromatography or preparative tlc eluting with 4 % meoh in dcm . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 83 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 90 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 45 ( m , 2h ), the title compounds were prepared according to the method described for example 1 using racemic n -[ 2 -( 1 -([ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethyl - silyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino ) ethyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 84 ). the residue was purified using silica gel column chromatography eluting with 6 % meoh in dcm followed by chiral separation using chiral preparative hplc to afford the separated enantiomers . fraction 1 : 44 mg , 100 % ee , registered as ( r )— enantiomer example 38 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 . 53 ( d , 3h ), 2 . 21 ( m , 2h ), 2 . 83 ( s , 3h ), 2 . 90 ( d , 3h ), 3 . 08 ( s , 3h ), 5 . 47 ( m , 1h ), 6 . 50 ( s , 1h ), 6 . 73 ( m , 2h ), 7 . 21 - 7 . 45 ( m , 4h ), 8 . 82 ( m , 1h ), 9 . 69 ( br s , 1h ), 9 . 86 ( m , 1h ), 13 . 57 ( br s , 1h ). ms m / z 541 [ m + h ] + fraction 2 : 41 mg , 87 . 5 % ee , registered as ( s )— enantiomer example 39 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 . 53 ( d , 3h ), 2 . 21 ( m , 2h ), 2 . 83 ( s , 3h ), 2 . 90 ( d , 3h ), 3 . 08 ( s , 3h ), 5 . 47 ( m , 1h ), 6 . 50 ( s , 1h ), 6 . 73 ( m , 2h ), 7 . 21 - 7 . 45 ( m , 4h ), 8 . 82 ( m , 1h ), 9 . 69 ( br s , 1h ), 9 . 86 ( m , 1h ), 13 . 57 ( br s , 1h ). ms m / z 541 [ m + h ] + to a solution of n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ]- n - methylbenzenesulfonamide ( preparation 81 , 400 mg , 0 . 44 mmol ) in meoh ( 2 ml ) was added dbu ( 0 . 20 ml , 1 . 31 mmol ), palladium acetate ( 6 . 85 mg , 0 . 03 mmol ) and molybdenum hexacarbonyl ( 115 mg , 0 . 44 mmol ) and the reaction was heated to 125 ° c . under microwave irradiation for 20 minutes . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography eluting with 70 % etoac in hexanes . the residue was dissolved in thf ( 4 ml ) and cooled to − 20 ° c . nmm ( 0 . 021 ml , 0 . 19 mmol ) followed by isobutylchloroformate ( 0 . 03 ml , 0 . 19 mmol ) were added and the reaction stirred at this temperature for 2 hours . aqueous ammonia was then added and the reaction stirred at room temperature for 1 hour . the reaction was quenched by the addition of water and extracted into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 40 % etoac in hexanes . the residue was treated with tfa ( 2 ml ) and stirred for 2 hours before concentrating in vacuo . ethylene diamine ( 0 . 5 ml ) was added and the reaction stirred at room temperature for 1 hour before concentrating in vacuo , pouring onto ice - water and extracting into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc eluting with 60 % etoac in hexanes to afford the title compound ( 23 mg , 10 % over 3 steps ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 92 ( t , 3h ), 3 . 04 ( s , 3h ), 3 . 40 ( m , 2h ), 4 . 70 ( br m , 1h ), 5 . 00 ( br m , 1h ), 6 . 55 ( d , 1h ), 6 . 64 ( s , 1h ), 6 . 80 ( d , 1h ), 7 . 06 ( m , 1h ), 7 . 14 ( m , 1h ), 7 . 29 ( m , 1h ), 7 . 47 ( m , 1h ), 7 . 59 - 7 . 67 ( m , 4h ), 7 . 73 ( m , 1h ), 7 . 83 ( br s , 1h ), 8 . 18 ( br s , 1h ), 9 . 65 ( m , 1h ), 9 . 77 ( s , 1h ), 13 . 60 ( br s , 1h ). ms m / z 575 [ m + h ] + a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethyl - silyl ) ethoxy ] methoxy } phenyl ]- 4 -({ 4 - methoxy - 2 -[ methyl ( methylsulfonyl ) amino ]- benzyl }- amino )- 1 -{[ 2 -( trimethylsilyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 21 , 102 mg , 0 . 15 mmol ) in tfa ( 5 ml ) was stirred at room temperature for 30 minutes before concentrating in vacuo . ethylene diamine ( 0 . 5 ml ) was added and the reaction stirred at room temperature for 15 minutes before concentrating in vacuo , pouring onto ice - water and extracting into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc to afford a white solid . the solid was dissolved in dcm ( 5 ml ) and boron tribromide ( 0 . 108 ml , 1 . 14 mmol ) was added dropwise at 0 ° c . and stirred at room temperature for 2 hours . the reaction was washed with saturated aqueous sodium bicarbonate solution , the organic layer collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc eluting with 5 % meoh in dcm to afford the title compound ( 37 mg , 54 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 02 ( s , 3h ), 3 . 08 ( s , 3h ), 3 . 79 ( m , 2h ), 4 . 65 ( br m , 2h ), 6 . 69 - 6 . 73 ( m , 2h ), 6 . 83 ( m , 1h ), 6 . 99 ( d , 1h ), 7 . 20 - 7 . 25 ( m , 2h ), 7 . 81 ( s , 1h ), 8 . 17 ( s , 1h ), 9 . 59 ( m , 2h ), 10 . 10 ( br s , 1h ), 13 . 70 ( br s , 1h ). ms m / z 583 [ m + h ] + the following examples ( examples 42 - 54 ) were prepared according to the method described for example 41 using the appropriate pyrazolo - pyridine , and purification method ( pm ) as described below if different from the method described : purification method g : silica gel column chromatography eluting with 5 - 7 % meoh in dcm followed by preparative hplc . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 98 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 87 ( m , 6h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 46 ( m , 2h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 83 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 45 ( m , 2h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( d , 3h ), a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- n - methyl - 4 -[({ 3 -[ methyl ( methylsulfonyl ) amino ] pyridin - 2 - yl } methyl ) amino ]- 1 -{[ 2 -( trimethylsilyl )- ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 43 , 110 mg , 0 . 13 mmol ) in tfa ( 5 ml ) was stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh and cooled in ice - water . ethylene diamine was added dropwise and stirred for 1 hour . the reaction was quenched by the addition of water and extracted into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative hplc to afford the title compound ( 18 mg , 26 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 84 ( d , 3h ), 3 . 07 ( s , 6h ), 3 . 61 ( m , 2h ), 4 . 70 - 5 . 20 ( br m , 2h ), 6 . 66 ( s , 1h ), 6 . 94 ( d , 1h ), 7 . 16 ( d , 1h ), 7 . 37 ( m , 1h ), 7 . 92 ( m , 1h ), 8 . 47 ( m , 1h ), 9 . 77 ( m , 1h ), 10 . 08 ( br s , 1h ), 13 . 63 ( br s , 1h ). ms m / z 582 [ m + h ] + the following examples ( examples 56 - 73 ) were prepared according to the method described for example 55 using the appropriate pyrazolo - pyridine and 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 89 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 83 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 85 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 84 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 84 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 08 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 09 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 84 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 84 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 00 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 81 ( m , 2h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 21 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 84 ( d , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 58 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 58 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 87 ( t , 3h ), the title compound was prepared according to the method described for example 55 using 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethylsilyl ) ethoxy )- methoxy ) phenyl )- n - methyl - 4 -(( 2 -( n - methyl - 2 - oxoxazolidine - 3 - sulfonamido ) benzyl )- amino )- 1 -(( 2 -( trimethylsilyl )- ethoxy )- methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 58 ). the residue was treated with 6m naoh ( 0 . 5 ml ) at 0 ° c . and stirred at room temperature for 18 hours . the reaction was acidified with hcl at 0 ° c ., and the resulting precipitate was filtered , extracted into etoac and concentrated in vacuo . the residue was purified using preparative tlc . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 07 ( m , 3h ), 2 . 84 ( s , 3h ), 3 . 02 ( s , 3h ), 3 . 39 ( m , 2h ), 3 . 69 ( m , 2h ), 4 . 70 ( m , 2h ), 4 . 90 ( br m , 2h ), 6 . 94 ( s , 1h ), 6 . 96 ( m , 1h ), 7 . 20 - 7 . 46 ( m , 5h ), 8 . 82 ( m , 1h ), 9 . 71 ( m , 1h ), 10 . 10 ( br s , 1h ), 13 . 70 ( s , 1h ). ms m / z 626 [ m + h ] + to a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl ]- 4 -({ 5 - methoxy - 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } amino )- 1 -{[ 2 -( trimethyl - silyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxylic acid ( preparation 11 , 100 mg , 0 . 12 mmol ) in dcm ( 3 ml ) was added boron tribromide ( 0 . 08 ml , 0 . 82 mmol ) and the reaction was stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo and triturated with ether / pentane . the resulting solid was dissolved in dmf ( 2 ml ) and 2 -[( 5 - aminopyridin - 2 - yl ) amino ] ethanol ( 51 mg , 0 . 33 mmol ) followed by dipea ( 0 . 07 ml , 0 . 17 mmol ) were added . hatu ( 159 mg , 0 . 42 mmol ) was added and the reaction stirred at room temperature for 18 hours . the reaction was partitioned between etoac and water , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc to afford the title compound ( 15 mg , 13 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 98 ( s , 3h ), 3 . 03 ( s , 3h ), 3 . 32 ( m , 2h ), 3 . 52 ( m , 2h ), 3 . 64 ( m , 2h ), 4 . 60 ( m , 1h ), 4 . 71 ( m , 1h ), 4 . 83 ( m , 1h ), 6 . 45 ( m , 1h ), 6 . 53 ( d , 1h ), 6 . 63 ( dd , 1h ), 6 . 75 ( s , 1h ), 6 . 78 ( m , 1h ), 6 . 96 ( d , 1h ), 7 . 21 - 7 . 26 ( m , 2h ), 7 . 78 ( dd , 1h ), 8 . 31 ( s , 1h ), 9 . 46 ( m , 2h ), 10 . 10 ( s , 1h ), 10 . 48 ( s , 1h ), 13 . 86 ( s , 1h ). ms m / z 719 [ m + h ] + to a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy }- phenyl ]- 4 -({ 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } amino )- 1 -{[ 2 -( trimethyl - silyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxylic acid ( preparation 12 , 100 mg , 0 . 12 mmol ) in dmf ( 3 ml ) was added 6 - methylpyridin - 3 - amine ( 65 mg , 0 . 60 mmol ), dipea ( 0 . 13 ml , 0 . 73 mmol ) and bop ( 267 mg , 0 . 60 mmol ) and the reaction was stirred at room temperature for 18 hours before concentrating in vacuo . the residue was partitioned between ice - water and etoac , the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 5 % meoh in dcm . the residue was treated with tfa ( 3 ml ) and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh and cooled in ice - water . ethylene diamine was added until the solution became basic , with stirring for 15 minutes . the solution was concentrated in vacuo and purified using preparative hplc to afford the title compound ( 35 mg , 29 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 45 ( s , 3h ), 3 . 04 ( s , 3h ), 3 . 11 ( s , 3h ), 3 . 62 ( m , 2h ), 4 . 80 ( m , 1h ), 4 . 95 ( m , 1h ), 6 . 77 ( s , 1h ), 6 . 96 ( d , 1h ), 7 . 20 - 7 . 33 ( m , 4h ), 7 . 41 ( m , 1h ), 7 . 49 ( m , 1h ), 8 . 13 ( dd , 1h ), 8 . 87 ( d , 1h ), 9 . 33 ( t , 1h ), 10 . 85 ( br s , 1h ). ms m / z 658 [ m + h ] + 6 -( 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- n -( tert - butyl )- 4 -(( 2 -( n - methylmethylsulfonamido ) benzyl ) amino )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 27 , 60 mg , 0 . 07 mmol ) was treated with tfa ( 8 ml ) and heated to reflux for 18 hours . the reaction was cooled , concentrated in vacuo , quenched by the addition of saturated aqueous sodium bicarbonate solution and extracted into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the crude residue was purified using preparative tlc to afford the title compound ( 21 mg , 51 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 04 ( s , 3h ), 3 . 14 ( s , 3h ), 3 . 69 ( m , 2h ), 4 . 70 ( br m , 1h ), 4 . 90 ( m , 1h ), 6 . 70 ( s , 1h ), 6 . 96 ( d , 1h ), 7 . 22 ( d , 1h ), 7 . 27 - 7 . 34 ( m , 2h ), 7 . 40 ( m , 1h ), 7 . 49 ( m , 1h ), 7 . 85 ( br s , 1h ), 8 . 20 ( br s , 1h ), 9 . 75 ( m , 1h ), 10 . 09 ( s , 1h ), 13 . 70 ( s , 1h ). ms m / z 567 [ m + h ] + 6 -( 4 - benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- n -( tert - butyl )- 4 -((( 5 - fluoro - 2 -( n - meth - ylmethylsulfonamido ) pyridin - 3 - yl ) methyl ) amino )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo -[ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 55 , 80 mg , 0 . 10 mmol ) was treated with tfa ( 10 ml ) and heated to reflux for 18 hours . the reaction was cooled , concentrated in vacuo and partitioned between saturated aqueous sodium bicarbonate solution and etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc to afford the title compound ( 20 mg , 35 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 3h ), 3 . 12 ( s , 3h ), 3 . 57 ( q , 2h ), 4 . 80 ( m , 2h ), 6 . 75 ( s , 1h ), 6 . 95 ( d , 1h ), 7 . 18 ( d , 1h ), 7 . 62 ( dd , 1h ), 7 . 89 ( br s , 1h ), 8 . 24 ( br s , 1h ), 8 . 42 ( m , 1h ), 9 . 85 ( m , 1h ), 10 . 11 ( br s , 1h ), 13 . 73 ( br s , 1h ). ms m / z 586 [ m + h ] + the following examples ( examples 79 - 91 ) were prepared according to the method described for example 78 using the appropriate pyrazolo - pyridine . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 20 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 90 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 89 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 01 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 19 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 22 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 86 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 05 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 05 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 88 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 90 ( t , 3h ), 6 -( 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- n -( tert - butyl )- 4 -(( 5 - methoxy - 2 -( n - methylmethylsulfonamido ) benzyl ) amino )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 29 , 100 mg , 0 . 12 mmol ) was heated to 100 ° c . in neat tfa ( 15 ml ) for 18 hours . the reaction was cooled , concentrated in vacuo and partitioned between saturated aqueous sodium bicarbonate solution and etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 55 % etoac in hexanes . the residue was stirred with neat boron tribromide ( 8 eq ) at 0 ° c . for 4 hours . the reaction was partitioned between dcm and saturated aqueous sodium bicarbonate solution , the organic layer collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc eluting with 5 % meoh in dcm to afford the title compound ( 30 mg , 51 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 98 ( s , 3h ), 3 . 04 ( s , 3h ), 3 . 66 ( m , 2h ), 4 . 58 ( m , 1h ), 4 . 79 ( m , 1h ), 6 . 63 ( dd , 1h ), 6 . 70 ( s , 1h ), 6 . 78 ( m , 1h ), 6 . 93 ( d , 1h ), 7 . 19 - 7 . 26 ( m , 2h ), 7 . 85 ( br s , 1h ), 8 . 20 ( br s , 1h ), 9 . 50 ( s , 1h ), 9 . 70 ( m , 1h ), 10 . 08 ( s , 1h ), 13 . 67 ( s , 1h ). ms m / z 583 [ m + h ] + to a solution of n -( 2 -((( 6 -( 4 -( benzyloxy )- 2 - ethyl - 5 - fluorophenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 272 , 249 mg , 0 . 36 mmol ) and methylamine in thf ( 3 ml , 2m ) was added molybdenum hexacarbonyl ( 96 mg , 0 . 36 mmol ) and palladium acetate ( 5 . 7 mg , 0 . 025 mmol ) followed by dbu ( 165 mg , 1 . 09 mmol ) and the reaction was heated to 100 ° c . under microwave irradiation for 10 minutes . the reaction was concentrated in vacuo , diluted with etoac and filtered through celite . the filtrate was concentrated in vacuo and purified using silica gel column chromatography followed by preparative tlc . the residue was dissolved in ethanol ( 7 ml ) and hydrogenated with pd ( oh ) 2 ( 15 mg ) at 40 psi for 16 hours . the reaction was filtered through celite and concentrated in vacuo . the residue was triturated with pentane and ether to afford the title compound as an off white solid ( 42 mg , 65 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 98 ( t , 3h ), 2 . 83 ( m , 4h ), 3 . 07 ( s , 3h ), 3 . 18 ( s , 3h ), 3 . 39 ( m , 1h ), 4 . 90 ( br m , 1h ), 5 . 05 ( br m , 1h ), 6 . 82 ( m , 1h ), 7 . 35 - 7 . 56 ( m , 5h ), 8 . 86 ( m , 1h ), 9 . 81 ( t , 1h ), 10 . 02 ( br s , 1h ), 13 . 97 ( br s , 1h ). ms m / z 528 [ m + h ] + the title compound was prepared according to the method described for preparation 93 using n -( 2 -((( 6 -( 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 276 ). the residue was purified using silica gel column chromatography eluting with 60 % etoac in hexanes . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 82 ( d , 3h ), 3 . 07 ( s , 3h ), 3 . 17 ( s , 3h ), 4 . 27 ( m , 2h ), 4 . 91 - 5 . 01 ( br m , 2h ), 6 . 97 ( m , 1h ), 7 . 32 - 7 . 41 ( m , 3h ), 7 . 54 ( m , 1h ), 7 . 67 ( m , 1h ), 8 . 87 ( t , 1h ), 9 . 91 ( t , 1h ), 10 . 37 ( br s , 1h ), 14 . 03 ( br s , 1h ). ms m / z 582 [ m + h ] + to a solution of 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 4 -(( 2 -( n - methylphenylsulfonamido ) benzyl ) amino )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo -[ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 258 , 300 mg , 837 . 14 mmol ) in anhydrous thf ( 10 ml ) was added nmm ( 0 . 06 ml , 0 . 57 mmol ) and isobutylchloroformate ( 0 . 07 ml , 0 . 57 mmol ) at − 20 ° c . and the reaction mixture was stirred at this temperature for 2 hours . aqueous ammonia ( 0 . 6 ml ) was added and the reaction stirred at room temperature for 1 hour . the reaction was quenched by the addition of water and etoac . the organic layer was separated , washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 66 % etoac in hexanes . the residue was dissolved in tfa and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo and dissolved in meoh ( 5 ml ), cooling to 0 ° c . ethylene diamine was added drop - wise until the solution showed a basic ph . the reaction was extracted into 20 % ipa in dcm , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc to afford the title compound ( 30 mg , 27 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 96 ( t , 3h ), 2 . 89 ( q , 2h ), 3 . 10 ( s , 3h ), 4 . 90 ( br m , 1h ), 5 . 08 ( br m , 1h ), 6 . 57 ( m , 1h ), 6 . 81 ( m , 1h ), 7 . 19 ( m , 1h ), 7 . 32 ( m , 1h ), 7 . 46 ( m , 1h ), 7 . 55 - 7 . 67 ( m , 5h ), 7 . 73 ( m , 1h ), 7 . 91 ( br s , 1h ), 8 . 26 ( br s , 1h ), 9 . 86 ( t , 1h ), 10 . 05 ( br s , 1h ), 13 . 96 ( br s , 1h ). ms m / z 575 [ m + h ] + the title compound was prepared according to the method described for example 95 using 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethylsilyl ) ethoxy ) meth - oxy ) phenyl )- 4 -(( 2 -( n - meth - ylphenylsulfonamido ) benzyl ) amino )- 1 -(( 2 -( trimethylsilyl ) eth - oxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 261 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 10 ( s , 3h ), 4 . 32 ( m , 2h ), 4 . 92 ( m , 1h ), 5 . 06 ( m , 1h ), 6 . 56 ( m , 1h ), 6 . 99 ( m , 1h ), 7 . 17 ( m , 1h ), 7 . 32 ( m , 1h ), 7 . 44 ( m , 1h ), 7 . 61 - 7 . 67 ( m , 6h ), 7 . 93 ( br s , 1h ), 8 . 28 ( br s , 1h ), 9 . 93 ( t , 1h ), 10 . 41 ( br s , 1h ), 14 . 04 ( br s , 1h ). ms m / z 630 [ m + h ] + to a solution of 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 4 -(( 5 - methoxy - 2 -( n - methylmethylsulfonamido ) benzyl ) amino )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo -[ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 269 , 0 . 1 g , 0 . 13 mmol ), hobt ( 36 mg , 0 . 27 mmol ) and edcl ( 51 mg , 0 . 27 mmol ) in dichloromethane ( 6 ml ) at 0 ° c . was added ammonium chloride ( 36 mg , 0 . 67 mmol ) and dipea ( 0 . 12 ml , 0 . 67 mmol ) and the reaction was stirred at room temperature for 14 hours . the reaction was concentrated in vacuo and the residue diluted with ethyl acetate . the organic solution was washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 52 % etoac in hexanes . the residue ( 62 mg , 0 . 084 mmol ) was dissolved in dcm ( 5 ml ) at 0 ° c . and boron tribromide ( 0 . 08 ml , 0 . 83 mmol ) was added . the reaction was stirred at room temperature for 1 hour . the reaction was concentrated in vacuo , diluted with methanol ( 5 ml ) and treated with ethylene diamine until the ph was basic , stirring for 1 hour . the solvent was removed in vacuo and the residue was partitioned between ethyl acetate and water , the organic extracts were dried over sodium sulfate and purified by preparative tlc to afford the title compound as an off - white solid ( 25 mg , 51 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 01 ( s , 3h ), 3 . 12 ( s , 3h ), 4 . 30 ( m , 2h ), 4 . 80 ( m , 1h ), 4 . 91 ( m , 1h ), 6 . 68 - 6 . 71 ( m , 1h ), 6 . 76 - 6 . 77 ( m , 1h ), 6 . 97 - 7 . 00 ( m , 1h ), 7 . 31 - 7 . 33 ( m , 1h ), 7 . 69 - 7 . 72 ( m , 1h ), 7 . 93 ( br s , 1h ), 8 . 28 ( br s , 1h ), 9 . 61 ( br s , 1h ), 9 . 87 ( t , 1h ), 10 . 40 ( br s , 1h ). ms m / z 584 [ m + h ] + the title compound was prepared according to the method described by example 97 using 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 4 -(( 2 -( n - methylmethylsulfonamido )- benzyl ) amino )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 270 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 3h ), 3 . 18 ( s , 3h ), 4 . 29 ( m , 2h ), 4 . 88 - 5 . 00 ( br m , 2h ), 6 . 98 ( m , 1h ), 7 . 32 - 7 . 43 ( m , 3h ), 7 . 54 ( m , 1h ), 7 . 68 ( m , 1h ), 7 . 92 ( br s , 1h ), 8 . 27 ( br s , 1h ), 9 . 92 ( t , 1h ), 10 . 39 ( br s , 1h ), 14 . 02 ( br s , 1h ). ms m / z 568 [ m + h ] + to a solution of 6 -( 5 - fluoro - 4 - hydroxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 4 -(( 5 - hydroxy - 2 -( n - methyl methylsulfonamido ) benzyl ) amino )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( example 174 , 160 mg , 0 . 27 mmol ) and 2 -[( 5 - aminopyridin - 2 - yl ) amino ] ethanol ( 84 mg , 0 . 54 mmol ) in dmf ( 5 ml ) was added hatu ( 312 mg , 0 . 82 mmol ) and dipea ( 0 . 12 ml , 0 . 68 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was purified directly by preparative hplc to afford the title compound ( 48 mg , 24 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 02 ( s , 3h ), 3 . 12 ( s , 3h ), 3 . 32 ( m , 2h ), 3 . 51 ( m , 2h ), 4 . 30 ( m , 2h ), 4 . 81 - 4 . 93 ( m , 2h ), 6 . 44 - 6 . 52 ( m , 2h ), 6 . 68 ( m , 1h ), 6 . 76 ( m , 1h ), 6 . 97 ( m , 1h ), 7 . 33 ( m , 1h ), 7 . 69 - 7 . 77 ( m , 2h ), 8 . 31 ( d , 1h ), 9 . 63 ( t , 1h ), 10 . 53 ( br s , 1h ). ms m / z 720 [ m + h ] + to a solution of n -( 3 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) pyrazin - 2 - yl )- n - methylmethanesulfonamide ( preparation 277 , 220 mg , 0 . 33 mmol ) in methylamine in thf ( 3 ml ) was added molybdenum hexacarbonyl ( 87 . 77 mg , 0 . 33 mmol ), pd ( oac ) 2 ( 5 . 18 mg , 0 . 07 mmol ) and dbu ( 0 . 15 ml , 0 . 99 mmol ) and the reaction was heated to 100 ° c . under microwave irradiation for 10 minutes . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography followed by preparative tlc . the residue was dissolved in dcm ( 5 ml ) and treated with boron tribromide ( 0 . 11 ml , 1 . 17 mmol ) at 0 ° c . and stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and partitioned between dcm and saturated aqueous sodium bicarbonate solution . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc eluting with 5 % meoh in dcm to afford the title compound as a white solid ( 23 mg , 47 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 84 ( s , 3h ), 3 . 15 ( s , 3h ), 3 . 19 ( s , 3h ), 4 . 21 ( m , 2h ), 5 . 13 ( m , 2h ), 6 . 97 ( m , 1h ), 7 . 58 ( m , 1h ), 8 . 53 ( d , 1h ), 8 . 63 ( d , 1h ), 8 . 86 ( m , 1h ), 10 . 06 ( m , 1h ), 10 . 37 ( br s , 1h ), 14 . 04 ( br s , 1h ). ms m / z 584 [ m + h ] + the following examples ( examples 101 - 104 ) were prepared according to the method described for example 1 using the appropriate pyrazolo - pyrimidine as described below : 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 10 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 83 ( m , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 00 ( t , 3h ), the title compound was prepared according to the method described for example 32 using n - ethyl - n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1 -(( 2 -( trimethyl - silyl ) ethoxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino )- methyl )- 4 - methoxyphenyl )- benzenesulfonamide ( preparation 266 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 93 ( t , 3h ), 2 . 85 ( d , 3h ), 3 . 24 ( m , 1h ), 3 . 77 ( m , 1h ), 4 . 29 ( m , 2h ), 4 . 85 - 4 . 93 ( m , 2h ), 6 . 36 ( m , 1h ), 6 . 54 ( m , 1h ), 6 . 76 ( m , 1h ), 6 . 98 ( m , 1h ), 7 . 58 - 7 . 72 ( m , 5h ), 8 . 89 ( m , 1h ), 9 . 62 ( s , 1h ), 9 . 85 ( br s , 1h ), 10 . 10 ( br s , 1h ), 14 . 10 ( br s , 1h ). ms m / z 672 [ m − h ] − the title compound was prepared according to the method described for example 32 using n - methyl - n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1 -(( 2 -( trimethyl - silyl ) ethoxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl )- amino ) methyl )- 4 - methoxyphenyl )- benzenesulfonamide ( preparation 267 ). 1 h nmr ( 400 mhz , dmso - dc ): δ ppm 2 . 85 ( s , 3h ), 3 . 05 ( s , 3h ), 4 . 34 ( m , 2h ), 4 . 86 ( m , 1h ), 4 . 97 ( m , 1h ), 6 . 32 ( m , 1h ), 6 . 51 ( m , 1h ), 6 . 76 ( m , 1h ), 6 . 98 ( m , 1h ), 7 . 60 - 7 . 74 ( m , 6h ), 8 . 91 ( m , 1h ), 9 . 63 ( s , 1h ), 9 . 89 ( t , 1h ), 10 . 40 ( br s , 1h ), 14 . 10 ( br s , 1h ). ms m / z 660 [ m + h ] + the following examples ( examples 107 - 108 ) were prepared according to the method described for example 41 using the appropriate pyrazolo - pyrimidine as described below : 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 94 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 05 ( s , 3h ), to a solution of tert - butyl 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 4 -(( 2 -( methylamino ) benzyl ) amino )- 3 -( methylcarbamoyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 1 - carboxylate ( preparation 256 , 50 mg , 0 . 08 mmol ) in anhydrous thf ( 5 ml ) was added sodium hydride ( 3 mg , 0 . 08 mmol ) at 0 ° c . the reaction was stirred for 10 minutes before the addition of sulfamoyl chloride ( 7 mg , 0 . 06 mmol ) and further stirring at 0 ° c . for 2 . 5 hours . the reaction was quenched by the addition of ice - water and extracted into etoac . the organic layer was washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc and dissolved in dcm ( 5 ml ). the solution was treated with boron tribromide ( 0 . 08 ml , 0 . 8 mmol ) and stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and partitioned between dcm and saturated aqueous sodium bicarbonate solution . the organic layer was washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc to afford the title compound as a white solid ( 11 mg , 32 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 86 ( s , 3h ), 3 . 08 ( s , 3h ), 4 . 29 ( m , 2h ), 4 . 86 - 5 . 05 ( m , 2h ), 6 . 97 ( m , 1h ), 7 . 09 ( s , 2h ), 7 . 25 - 7 . 35 ( m , 3h ), 7 . 46 ( m , 1h ), 7 . 69 - 7 . 72 ( m , 1h ), 8 . 88 ( t , 1h ), 9 . 88 ( t , 1h ), 10 . 37 ( s , 1h ), 14 . 02 ( s , 1h ). ms m / z 583 [ m + h ] + to a solution of tert - butyl 6 -( 4 -(( tert - butoxycarbonyl ) oxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 4 -(( 2 -( methylamino ) benzyl ) amino )- 3 -( methylcarbamoyl )- 1h - pyrazolo -[ 3 , 4 - d ] pyrimidine - 1 - carboxylate ( preparation 255 , 56 mg , 0 . 08 mmol ) in thf ( 3 ml ) was added sodium hydride ( 2 mg , 0 . 08 mmol ) at 0 ° c . the reaction was stirred at room temperature for 2 minutes before the addition of methanesulfonyl chloride ( 10 mg , 0 . 08 mmol ) and further stirring for 18 hours . the reaction was quenched by the addition of ice - water and extracted into etoac . the organic layer was washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc and treated with 4m hcl in dioxane ( 0 . 3 ml ). the reaction was stirred at room temperature for 3 hours before concentrating in vacuo and triturating with pentane - ether to afford the title compound . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 62 ( d , 3h ), 2 . 86 ( d , 3h ), 3 . 05 ( s , 3h ), 4 . 33 ( m , 2h ), 4 . 95 ( m , 2h ), 6 . 98 ( m , 1h ), 7 . 28 - 7 . 49 ( m , 4h ), 7 . 73 ( m , 1h ), 8 . 89 ( t , 1h ), 9 . 92 ( t , 1h ), 10 . 38 ( s , 1h ), 14 . 03 ( s , 1h ). ms m / z 595 [ m − h ] − a 0 . 1 m solution of 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 328 , 700 μl , 70 μmol ) in toluene was added to an amine of formula ( xii ) ( 200 μmol , 2 . 9 eq ) and the solution degassed with nitrogen . cesium carbonate ( 45 mg , 140 μmol ) was added followed by pd 2 ( dba ) 3 ( 3 . 4 mg , 3 . 5 μmol ) and binap ( 2 . 2 mg , 3 . 5 μmol ) and the reaction further degassed with nitrogen . the reaction was shaken at 80 ° c . for 16 hours before concentrating in vacuo . water ( 1 ml ) was added followed by etoac ( 1 ml ) and the mixture filtered . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . to the residue was added a solution of chcl in etoh ( 1 ml , v : v 1 : 6 ) and the reaction shaken at 80 ° c . for 2 hours . the reaction was cooled , concentrated in vacuo and purified using one of the preparative hplc methods described below : method a : agella venusil asb c18 , 150 × 21 . 2 mm × 5 μm ; acetonitrile - water ( 0 . 225 % formic acid ); flow rate : 35 ml / min ; gradient time 8 mins . method b : boston symmetrix ods - h , 150 × 30 mm × 5 μm ; acetonitrile - water ( 0 . 225 % formic acid ); flow rate : 30 ml / min ; gradient time 10 mins . method c : dikma diamonsil ( 2 ) c18 , 200 × 20 mm × 5 μm ; acetonitrile - water ( 0 . 225 % formic acid ); flow rate : 30 ml / min ; gradient time 10 mins . a : 0 . 0375 % tfa in water ; b : 0 . 01875 % tfa in mecn ; column : xbridge c18 , 2 . 1 × 50 mm × 5 μm ; gradient : from 99 % [ a ] and 1 % [ b ] to 95 % [ a ] and 5 % [ b ] in 0 . 6 min , further to 100 % [ b ] in 4 . 0 min and finally back to initial condition in 4 . 3 min , 0 . 8 ml / min flow rate . a : 0 . 0375 % tfa in water ; b : 0 . 01875 % tfa in mecn ; column : xbridge c18 , 2 . 1 × 50 mm × 5 μm ; gradient : from 90 % [ a ] and 10 % [ b ] to 100 % [ b ] in 4 min and finally back to initial condition in 4 . 3 min , 0 . 8 ml / min flow rate . a : 0 . 0375 % tfa in water ; b : 0 . 01875 % tfa in mecn ; column : xbridge c18 , 2 . 1 × 50 mm × 5 μm ; gradient : from 75 % [ a ] and 25 % [ b ] to 100 % [ b ] in 3 . 5 min and finally back to initial condition in 4 . 0 min , 0 . 8 ml / min flow rate . the compounds of the examples in the table below ( examples 111 - 124 ) were prepared and purified from 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 328 ) and the appropriate amine according to library protocol 1 . the compounds were isolated as their formate salts . to a solution of 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethyl - silyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 328 , 14 mg , 0 . 02 mmol ) in toluene ( 0 . 5 ml ) was added n -[ 2 - aminomethyl ) phenyl ]- n - methylmethanesulfonamide ( wo20101058846a1 , 9 mg , 0 . 03 mmol ), cesium carbonate ( 14 mg , 0 . 04 mmol ), pd ( oac ) 2 ( 0 . 9 mg , 0 . 004 mmol ) and binap ( 3 . 7 mg , 0 . 006 mmol ). the reaction was degassed with nitrogen followed by heating to 150 ° c . under microwave irradiation for 15 minutes . the reaction was filtered , washing through with dcm and the filtrate concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 1 - 5 % meoh in dcm . the residue was dissolved in meoh ( 1 ml ) and chcl ( 0 . 2 ml ) was added and the reaction heated to 80 ° c . for 3 hours . the reaction was cooled , concentrated in vacuo and triturated with dcm to afford the title compound as the hydrochloride salt ( 10 mg , 50 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 75 ( t , 3h ), 2 . 40 ( q , 2h ), 3 . 00 ( s , 3h ), 3 . 03 ( s , 3h ), 4 . 55 ( br m , 1h ), 4 . 95 ( br m , 1h ), 6 . 50 ( m , 3h ), 7 . 00 ( m , 1h ), 7 . 25 ( m , 2h ), 7 . 40 - 7 . 50 ( m , 2h ), 7 . 80 ( t , 1h ), 8 . 20 ( s , 1h ), 9 . 25 ( s , 1h ), 12 . 95 ( s , 1h ). ms m / z 452 [ m + h ] + the following compounds of the examples below ( examples 126 - 130 ) were prepared according to the method described for example 125 above using 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 328 ) or 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 331 ) or 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesuffonate ( preparation 330 ) and the appropriate amine . purification took place according to the purification method ( pm ) described or one of the following below . compounds were isolated as the free parent , diethylamine salt or hydrochloride salt as described below : purification method j : the residue was dissolved in dmso ( 0 . 9 ml ) and triethylamine ( 0 . 1 ml ) and purified using preparative hplc . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 85 ( br s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 94 - 1 . 06 ( m , to a stirring solution of 4 - nitrophenyl { 2 -[ methyl ( methyl - sulfonyl ) amino ] benzyl } carbamate ( preparation 166 , 2 . 02 g , 5 . 32 mmol ) and triethylamine ( 2 . 12 ml , 15 . 33 mmol ) in anhydrous dmf ( 20 ml ) was added 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 332 , 1 . 5 g , 3 . 07 mmol ) and the reaction was heated to 80 ° c . for 15 hours . the reaction was concentrated in vacuo and partitioned between water and ethyl acetate . the organic layer was washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 1 % meoh in dcm . the residue was dissolved in meoh ( 10 ml ) and chcl ( 8 ml ) was added . the reaction was heated at from 65 - 80 ° c . for 6 hours before cooling and concentrating in vacuo . the residue was triturated with mecn / ether to afford the title compound as the hydrochloride salt ( 400 mg , 42 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 98 ( m , 3h ), 2 . 32 - 2 . 41 ( m , 2h ), 3 . 09 - 3 . 18 ( m , 6h ), 5 . 11 ( m , 2h ), 6 . 98 ( s , 2h ), 7 . 09 - 7 . 21 ( m , 2h ), 7 . 21 ( s , 2h ), 7 . 27 ( br s , 21h ), 7 . 34 - 7 . 36 ( m , 1h ), 7 . 42 - 7 . 48 ( m , 3h ), 7 . 63 ( br s , 1h ), 8 . 70 ( br s , 1h ), 10 . 04 ( br s , 1h ), 10 . 35 ( br s , 1h ), 12 . 20 ( br s , 1h ), 14 . 17 ( br s , 1h ). ms m / z 470 [ m + h ] + the following compounds of the examples below ( examples 132 - 136 ) were prepared according to the method described for example 131 above using 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 332 ) or 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl )- ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 333 ) and the appropriate aminocarbamate . the compounds were isolated as their hydrochloride salts . purification was carried out according to the purification method ( pm ) described or one of the following below : 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 00 ( s , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 87 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 05 ( t , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 98 ( m , 3h ), 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 04 ( m , 5h ), to 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimeth - ylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 331 , 200 mg , 0 . 30 mmol ) and n -( 2 -( aminomethyl ) phenyl )- n - propylmethanesulfonamide trifluoroacetate ( preparation 349 , 172 mg , 0 . 71 mmol ) in dmf ( 5 ml ) was added triethylamine ( 0 . 19 ml , 1 . 4 mmol ) and the reaction was heated 10 to 110 ° c . for 2 hours . the reaction was cooled and partitioned between etoac and water . the organic layer was collected , washed with water , dried over magnesium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 1 : 1 etoac : heptanes . the residue was dissolved in meoh ( 5 ml ) and chcl ( 1 . 5 ml ) was added and the reaction heated to 60 ° c . for 18 hours . the reaction was cooled , concentrated in vacuo and purified using preparative hplc to afford the title compound as the free parent . rt = 2 . 85 minutes ; ms m / z 498 [ m + h ] + the following compounds of the examples below ( examples 138 - 144 ) were prepared according to the method described for example 137 above using 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo -[ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 331 ) and the appropriate amine . deprotection was carried out as described or using tfa / tes in place of chcl . purification was carried out according to the purification method ( pm ) described or one of the following below . the compounds were all isolated as free parents . purification method l : the reaction mixture was quenched by the addition of saturated aqueous nahco 3 solution and extracted into dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative hplc . to a solution of 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 330 , 400 mg , 0 . 64 mmol ) and n - ethyl - n -( 2 -(( methyl - amino ) methyl ) phenyl ) methanesulfonamide ( preparation 358 , 234 mg , 0 . 96 mmol ) in toluene ( 8 ml ) was added cesium carbonate ( 420 mg , 1 . 29 mmol ) and the mixture was degassed with nitrogen for 5 minutes . pd ( oac ) 2 ( 16 mg , 0 . 064 mmol ) and binap ( 60 mg , 0 . 096 mmol ) were added and the reaction was heated to 140 ° c . under microwave irradiation for 30 minutes . the reaction was partitioned between water and ethyl acetate , the organic layer collected , washed with brine , dried over sodium sulfate , and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 36 % etoac in hexanes . the residue ( 147 mg , 0 . 22 mmol ) was dissolved in meoh ( 10 ml ) and chcl ( 10 ml ) was added with heating to 65 ° c . for 4 hours . the reaction was concentrated in vacuo and triturated with pentane - ether to afford the title compound as the hydrochloride salt ( 125 mg , 34 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 00 - 1 . 13 ( m , 6h ), 2 . 56 ( m , 2h ), 3 . 07 ( s , 3h ), 3 . 43 ( s , 3h ), 3 . 66 ( m , 2h ), 5 . 19 ( br s , 2h ), 6 . 82 ( s , 1h ), 6 . 88 ( d , 1h ), 7 . 09 ( d , 1h ), 7 . 23 ( t , 3h ), 7 . 33 ( t , 1h ), 7 . 39 ( t , 1h ), 7 . 53 ( d , 1h ), 8 . 16 ( s , 1h ), 10 . 31 ( br s , 1h ), 11 . 95 ( br s , 1h ), 14 . 16 ( br s , 1h ). ms m / z 496 [ m − h ] − the title compound was prepared according to the method described for example 145 using 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 330 ), n - methyl - n -( 2 -(( methylamino ) methyl ) phenyl ) methanesulfonamide ( preparation 359 ). the residue was triturated with pentane - ether and further purified by preparative tlc to afford the title compound as the hydrochloride salt ( 60 mg , 59 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 91 ( m , 3h ), 2 . 54 ( m , 2h ), 3 . 09 ( s , 3h ), 3 . 16 ( s , 3h ), 3 . 37 ( s , 3h ), 5 . 10 ( s , 2h ), 6 . 67 ( s , 1h ), 6 . 79 ( d , 1h ), 7 . 01 ( d , 1h ), 7 . 09 ( d , 1h ), 7 . 28 ( t , 1h ), 7 . 34 ( t , 1h ), 7 . 55 ( d , 1h ), 8 . 06 ( s , 1h ), 9 . 74 ( s , 1h ), 13 . 08 ( s , 1h ). ms m / z 482 [ m − h ] − to a solution of n -( 2 -( benzyloxy ) ethyl )- n -( 2 -((( 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ) methyl ) phenyl ) methanesulfonamide ( preparation 327 , 79 mg , 0 . 09 mmol ) in 1 : 1 meoh : etoh ( 10 ml ) was added ammonium formate ( 1 mg , 0 . 09 mmol ) followed by palladium hydroxide ( 4 mg ). the reaction was heated to 70 ° c . for 18 hours before cooling and filtering thought celite . the filtrate was concentrated in vacuo and partitioned between etoac and water . the organic layer was collected , washed with water and brine , dried over sodium sulfate and concentrated in vacuo . the residue was dissolved in dcm ( 3 ml ) and tfa ( 141 μl , 1 . 84 mmol ) followed by triethylsilane ( 21 . 5 μl , 0 . 18 mmol ) were added . the reaction was heated to 70 ° c . for 72 hours before cooling and quenching with saturated aqueous nahco 3 solution . the reaction was extracted into etoac , and the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was triturated with dcm to afford the title compound ( 16 mg , 43 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 90 ( t , 3h ), 2 . 25 - 2 . 40 ( m , 2h ), 3 . 18 ( s , 3h ), 3 . 25 ( m , 2h ), 3 . 75 - 3 . 80 ( m , 1h ), 3 . 83 - 3 . 95 ( m , 1h ), 4 . 75 ( m , 1h ), 5 . 30 ( m , 1h ), 6 . 70 ( s , 1h ), 6 . 80 ( m , 1h ), 6 . 95 ( m , 1h ), 7 . 40 ( m , 2h ), 7 . 50 ( m , 1h ), 7 . 63 ( m , 1h ), 8 . 28 ( s , 1h ). ms m / z 500 [ m + h ] + to a solution of 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 334 , 100 mg , 0 . 21 mmol ) in dcm ( 1 ml ) was added n -[ 2 -( aminomethyl ) phenyl ]- n - methylmethanesulfonamide ( wo20101058846a1 , 60 mg , 0 . 28 mmol ) followed by pybrop ( 130 mg , 0 . 28 mmol ) and dipea ( 0 . 14 ml , 0 . 81 mmol ). the reaction was stirred at room temperature for 18 hours . the reaction was poured into saturated aqueous nahco 3 solution and extracted with dcm three times . the organic layers were collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 1 : 1 etoac in hexanes . the residue was dissolved in dcm ( 0 . 6 ml ) and tfa ( 0 . 2 ml ) followed by tes ( 0 . 05 ml ) were added at 0 ° c . the reaction was stirred at room temperature for 18 hours before being quenched with saturated aqueous nahco 3 solution . the mixture was extracted into dcm , the organic layer collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 60 % etoac in heptanes . the residue was dissolved in dcm ( 1 ml ) and bbr 3 ( 0 . 72 ml ) was added at 0 ° c . the reaction was stirred at room temperature for 18 hours before concentrating in vacuo and purifying by preparative hplc to afford the title compound . ms m / z 524 [ m + h ] + rt = 2 . 39 minutes . the title compound was prepared according to the method described for example 148 using n - methyl - n -( 2 -(( methylamino ) methyl ) phenyl ) methanesulfonamide ( preparation 359 ). ms m / z 538 [ m + h ] + rt = 2 . 52 minutes . the title compound was prepared according to the method described for example 148 using the free base of n -[ 2 -( aminomethyl )- 4 - methylphenyl ]- n - methylmethanesulfonamide hydrochloride ( preparation 189 ). ms m / z 538 [ m + h ] + rt = 2 . 47 minutes . to a 0 . 2m solution of amines of formula ( xii ) ( 1 ml , 200 umol ) in nbuoh was added a 0 . 2m solution of 4 , 6 - dichloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine ( wo20131014567a1 , 1 ml , 200 umol ) followed by dipea ( 120 ul , 700 umol ). the reaction was heated to 80 ° c . for 16 hours before concentrating in vacuo . the residue was dissolved in 1 : 1 meoh : toluene ( 1 . 5 ml ). to the solution was added potassium carbonate ( 62 mg , 450 umol ), peppsi ™- ipr ( 3 mg , 4 . 5 umol ) and ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenoxy ] methoxy } ethyl )( trimethyl ) silane ( preparation 150 , 400 mmol ). the reaction was heated to 100 ° c . under microwave irradiation for 25 minutes before concentrating in vacuo . the residue was dissolved in etoac ( 5 ml ) and washed with water ( 3 ml ) and brine ( 3 ml ). the organic extract was dried over sodium sulfate and concentrated in vacuo . the residue was dissolved in tfa ( 1 ml ) and stirred at room temperature for 16 hours . the reaction was concentrated in vacuo and azeotroped with toluene . the residue was dissolved in meoh and ethylenediamine ( 35 ul , 500 umol ) was added with stirring at room temperature for 18 hours . the reaction was concentrated in vacuo , dissolved in dmso ( 1 ml ) and purified using preparative hplc as described below : a : 0 . 05 % formic acid in water ; b : mecn ; column : restek c18 , 30 × 2 . 1 mm × 3 μm ; gradient : from 98 % [ a ] and 2 % [ b ] to 90 % [ a ] and 10 % [ b ] in 1 min , further to 98 % [ b ] in 2 min and finally back to initial condition in 2 . 90 min , 1 . 5 ml / min flow rate . method a : gemini nxc18 ( 100 × 20 mm × 5μ ); acetonitrile - water ( 20 mm nh 4 co 3 ); flow rate 20 ml / min ; gradient time 10 mins for 10 - 75 % organic elution . method b : reprosil gold c18 ( 250 × 20 mm × 5μ ); acetonitrile - water ( 20 mm nh 4 co 3 ); flow rate 20 ml / min ; gradient time 18 mins for 10 - 70 % organic elution . the compounds of the examples in the table below ( examples 151 - 154 ) were prepared and purified from 4 , 6 - dichloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine ( wo20131014567a1 ), ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenoxy ] methoxy } ethyl )( trimethyl ) silane ( preparation 150 ) and the appropriate amine according to library protocol 2 . the title compound was prepared according to the method described for example 125 using n -( 2 -((( 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 312 ) and ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 - ethyl - phenoxy ] methoxy } ethyl )( trimethyl ) silane ( wo20131014567a1 ). sphos was used as the ligand and the final residue was triturated with pentane / ether to afford the hydrochloride salt . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 91 ( t , 3h ), 2 . 66 ( m , 2h ), 3 . 06 ( s , 3h ), 3 . 13 ( s , 3h ), 4 . 82 ( br s , 1h ), 5 . 06 ( br s , 1h ), 6 . 86 ( d , 1h ), 7 . 33 - 7 . 46 ( m , 4h ), 7 . 55 ( d , 1h ), 8 . 55 ( br s , 1h ), 10 . 33 ( br s , 1h ), 14 . 56 ( br s , 1h ). ms m / z 471 [ m + h ] + the title compound was prepared according to the method described for preparation 299 pyrimidines using 2 -( 4 -( benzyloxy )- 2 - ethyl - 6 - methylphenyl )- 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolane ( preparation 342 ) followed by treating the residue with tfa at reflux . the reaction was concentrated in vacuo and partitioned between etoac and saturated aqueous sodium bicarbonate solution . the organic extracts were washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative hplc . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 63 - 0 . 82 ( m , 3h ), 1 . 86 ( m , 5h ), 2 . 91 ( s , 3h ), 2 . 94 ( s , 3h ), 4 . 37 ( br m , 1h ), 4 . 85 ( br m , 1h ), 6 . 36 ( br m , 3h ), 7 . 20 ( m , 2h ), 7 . 33 ( m , 2h ), 8 . 17 ( m , 1h ), 9 . 03 ( m , 1h ), 12 . 88 ( br s , 1h ). ms m / z 466 [ m + h ] + a solution of n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 79 , 200 mg , 0 . 22 mmol ) and 2 - bromo - 1 -[[ 2 -( trimethylsilyl ) ethoxy ] methyl ]- 1h - imidazole ( j . org . chem . ( 2010 ) 75 ( 15 ) 4911 - 4920 , 62 . 102 mg , 0 . 22 mmol ) in toluene ( 2 ml ) was degassed with nitrogen for 5 minutes . bis ( tributyltin ) ( 0 . 27 ml , 0 . 54 mmol ) and copper ( i ) iodide ( 8 . 53 mg , 0 . 045 mmol ) were added followed by pd ( pph 3 ) 4 ( 25 . 88 mg , 0 . 022 mmol ) and the reaction was heated to 100 ° c . for 6 . 5 hours . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography eluting with 15 % etoac in hexanes . the residue ( 80 mg , 0 . 086 mmol ) was treated with tfa ( 2 ml ) and the solution stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh ( 5 ml ) and cooled in ice water . ethylene diamine was added dropwise until the solution was basic , with stirring for 15 minutes . the solution was concentrated in vacuo and purified using silica gel column chromatography eluting with 60 % etoac in hexanes to afford the title compound ( 25 mg , 54 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 91 ( t , 3h ), 2 . 43 ( m , 2h ), 3 . 06 ( s , 3h ), 3 . 10 ( s , 3h ), 4 . 70 ( br m , 1h ), 5 . 00 ( br m , 1h ), 6 . 58 ( m , 1h ), 6 . 77 ( m , 1h ), 7 . 05 ( m , 1h ), 7 . 10 ( s , 1h ), 7 . 26 - 7 . 31 ( m , 3h ), 7 . 47 ( m , 2h ), 9 . 74 ( s , 1h ), 10 . 89 ( t , 1h ), 12 . 93 ( s , 1h ), 13 . 29 ( s , 1h ). ms m / z 536 [ m + h ] + the title compound was prepared according to the method described by example 157 using n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 79 ) and 2 - bromo - 4 , 5 - dimethyl - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - imidazole ( preparation 386 ) at 115 ° c . under microwave irradiation for 30 minutes . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 97 ( t , 3h ), 2 . 56 ( m , 2h ), 3 . 04 ( s , 3h ), 3 . 10 ( s , 3h ), 4 . 60 ( br m , 1h ), 5 . 00 ( br m , 1h ), 6 . 55 ( s , 1h ), 6 . 79 ( m , 1h ), 7 . 08 ( m , 1h ), 7 . 35 ( m , 2h ), 7 . 49 ( m , 1h ), 7 . 51 ( m , 1h ), 9 . 74 ( s , 1h ), 10 . 95 ( t , 1h ), 12 . 40 ( s , 1h ), 13 / 15 ( s , 1h ). ms m / z 564 [ m + h ] + the title compound was prepared according to the method described by example 157 using n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 79 ) and 2 - iodo - 5 - methyl - 1h - imidazole at 115 ° c . under microwave irradiation for 30 minutes . following deprotection the residue was purified using preparative hplc . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 95 ( t , 1 . 5h ), 0 . 99 ( t , 1 . 5h ), 1 . 99 ( s , 1 . 5h ), 2 . 32 ( s , 1 . 5h ), 3 . 05 ( m , 3h ), 3 . 10 ( m , 3h ), 4 . 66 ( br m , 1h ), 4 . 99 ( br m , 1h ), 6 . 56 ( m , 1h ), 6 . 77 - 6 . 90 ( m , 2h ), 7 . 01 - 7 . 09 ( m , 1h ), 7 . 27 - 7 . 36 ( m , 2h ), 7 . 46 - 7 . 61 ( m , 2h ), 9 . 74 ( br s , 1h ), 10 . 95 ( m , 1h ), 12 . 55 ( s , 0 . 5h ), 12 . 69 ( s , 0 . 5h ), 13 . 22 ( br s , 1h ). ms m / z 550 [ m + h ] + the title compound was prepared according to the method described by example 157 using 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethylsilyl ) ethoxy )- methoxy ) phenyl )- 3 - iodo - n -( 2 -( methylthio ) ethyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - amine ( preparation 379 ) and tert - butyl 2 - iodo - 6 , 7 - dihydro - 1h - imidazo [ 4 , 5 - c ] pyridine - 5 ( 4h )- carboxylate ( wo20131014567a1 ). ms m / z 522 [ m + h ] + to a suspension of anhydrous magnesium sulfate ( 40 mg , 0 . 33 mmol ) and 2 - fluoro - 4 -( 4 -(( 2 -( methylthio ) ethyl ) amino )- 3 -( 4 , 5 , 6 , 7 - tetrahydro - 1h - imidazo [ 4 , 5 - c ] pyridin - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 6 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( example 161 , 44 mg , 0 . 08 mmol ) in methanol ( 2 . 5 ml ) was added a solution of benzaldehyde ( 0 . 017 ml , 0 . 17 mmol ) in methanol ( 2 . 5 ml ). the reaction was stirred for 1 hour at 55 ° c . before cooling to room temperature and adding sodium cyanoborohydride ( 10 . 6 mg , 0 . 17 mmol ). the reaction was stirred at room temperature for 18 hours . the reaction was filtered and concentrated in vacuo . the residue was partitioned between 20 % ipa in dcm and water . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc to afford the title compound ( 20 mg , 39 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 90 ( s , 1 . 5h ), 1 . 98 ( s , 1 . 5h ), 2 . 66 - 2 . 88 ( m , 6h ), 3 . 47 ( m , 2h ), 3 . 64 - 3 . 72 ( m , 4h ), 4 . 03 - 4 . 11 ( m , 2h ), 6 . 62 ( s , 1h ), 7 . 01 ( m , 1h ), 7 . 27 - 7 . 38 ( m , 6h ), 10 . 11 ( s , 1h ), 10 . 68 ( t , 0 . 5h ), 10 . 75 ( t , 0 . 5h ), 12 . 46 ( s , 0 . 5h ), 12 . 58 ( s , 0 . 5h ), 13 . 23 ( s , 1h ). ms m / z 612 [ m + h ] + to a solution of n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ]- n - methylmethanesufonamide ( preparation 79 , 250 mg , 0 . 29 mmol ) in dmso ( 0 . 5 ml ) was added pyrazole ( 19 . 88 mg , 0 . 29 mmol ), peg ( 500 mg ), cesium carbonate ( 133 mg , 0 . 41 mmol ), cuprous oxide ( 1 . 25 mg , 0 . 01 mmol ) and 4 , 7 - dimethoxy - 1 , 10 - phenanthroline ( 5 . 61 mg , 0 . 023 mmol ) and the reaction was heated to 110 ° c . for 18 hours . the reaction was cooled , diluted with etoac and washed with water and brine . the organic layer was dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 27 % etoac in hexanes . the residue ( 77 mg , 0 . 097 mmol ) was treated with tfa ( 2 . 5 ml ) and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh ( 5 ml ) and cooled in ice water . ethylene diamine was added dropwise until the solution was basic , with stirring for 15 minutes . the solution was concentrated in vacuo and purified using silica gel column chromatography eluting with 6 % meoh in dcm to afford the title compound ( 32 mg , 62 %). 1 h nmr ( 400 mhz , dmso - ds ): δ ppm 0 . 93 ( t , 3h ), 2 . 49 ( m , 2h ), 3 . 05 ( s , 3h ), 3 . 07 ( s , 3h ), 4 . 67 ( br m , 1h ), 5 . 00 ( br m , 1h ), 6 . 60 ( s , 1h ), 6 . 66 ( m , 1h ), 6 . 80 ( m , 1h ), 7 . 05 ( m , 1h ), 7 . 31 ( m , 2h ), 7 . 45 ( m , 2h ), 7 . 91 ( s , 1h ), 8 . 54 ( s , 1h ), 9 . 61 ( t , 1h ), 9 . 76 ( s , 1h ), 13 . 11 ( s , 1h ). ms m / z 536 [ m + h ] + the title compound was prepared according to the method described for example 162 using n - methyl - n -( 2 -{[( 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ]- methyl } phenyl ) methanesulfonamide ( preparation 105 ) and pyrazole . following deprotection the residue was purified using silica gel column chromatography eluting with 45 % etoac in hexanes . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 06 ( m , 6h ), 3 . 40 ( m , 2h ), 4 . 64 ( br m , 1h ), 4 . 93 ( br m , 1h ), 6 . 66 ( m , 2h ), 6 . 94 ( m , 1h ), 7 . 19 - 7 . 22 ( m , 1h ), 7 . 28 - 7 . 34 ( m , 2h ), 7 . 41 - 7 . 50 ( m , 2h ), 7 . 93 ( s , 1h ), 8 . 57 ( s , 1h ), 9 . 71 ( t , 1h ), 10 . 11 ( brs , 1h ), 13 . 21 ( brs , 1h ). ms m / z 590 [ m + h ] + to a solution of acetimidamide hydrochloride ( 33 mg , 0 . 35 mmol ) in 2 - methoxyethanol ( 3 ml ) was added dipea ( 0 . 087 ml , 0 . 50 mmol ) followed by n -( 2 -((( 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 3 -( hydrazinecarbonyl )- 1 -(( 2 -( tri - methylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino )- methyl ) phenyl )- n - methyl - methanesulfonamide ( preparation 374 , 120 mg , 0 . 143 mmol ). the reaction was stirred at 85 ° c . for 18 hours . the reaction was quenched with water and extracted with dcm . the organic extracts were collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 27 % etoac in dcm . the residue ( 100 mg , 0 . 12 mmol ) was treated with tfa ( 2 ml ) and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh ( 5 ml ) and cooled in ice water . ethylene diamine was added dropwise until the solution was basic , with stirring for 15 minutes . the solution was concentrated in vacuo and purified using preparative tlc to afford the title compound ( 40 mg , 54 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 75 ( s , 3h ), 3 . 04 - 3 . 12 ( m , 6h ), 3 . 68 ( m , 2h ), 4 . 70 ( br m , 1h ), 5 . 00 ( br m , 1h ), 6 . 67 ( m , 1h ), 6 . 97 ( m , 1h ), 7 . 20 - 7 . 22 ( m , 1h ), 7 . 25 - 7 . 29 ( m , 2h ), 7 . 34 ( m , 2h ), 7 . 85 ( m , 1h ), 10 . 08 - 10 . 25 ( m , 1h ), 13 . 36 ( s , 0 . 5h ), 13 . 71 ( s , 0 . 5h ), 14 . 02 ( s , 0 . 5h ), 14 . 52 ( s , 0 . 5h ). ms m / z 605 [ m + h ] + to a solution of 6 -[ 5 - fluoro - 2 - ethyl - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 4 -({ 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } amino )- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxylic acid ( preparation 373 , 180 mg , 0 . 23 mmol ) in thf ( 12 ml ) was added hydrazine hydrochloride ( 39 . 82 mg , 0 . 58 mmol ), bop ( 257 mg , 0 . 58 mmol ) and dipea ( 0 . 122 ml , 0 . 70 mmol ). the reaction was stirred at room temperature for 18 hours . the reaction was quenched with water , extracted into dcm , the organic extracts collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 8 % meoh in dcm . the residue was added to a solution of acetimidamide hydrochloride ( 30 mg , 0 . 32 mmol ) and dipea ( 0 . 08 ml , 0 . 44 mmol ) in 2 - methoxyethanol ( 3 ml ) and the reaction was heated to 85 ° c . for 18 hours . the reaction was cooled and quenched by the addition of water and extracted into dcm . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc . the residue was treated with tfa ( 2 ml ) and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh ( 5 ml ) and cooled in ice water . ethylene diamine was added dropwise until the solution was basic , with stirring for 15 minutes . the solution was concentrated in vacuo and purified using preparative tlc to afford the title compound ( 30 mg , 88 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 97 ( t , 3h ), 2 . 20 ( s , 3h ), 2 . 56 ( m , 2h ), 3 . 04 ( s , 3h ), 3 . 12 ( s , 3h ), 4 . 65 ( br m , 1h ), 5 . 00 ( br m , 1h ), 6 . 66 ( m , 1h ), 6 . 83 ( m , 1h ), 7 . 02 ( m , 1h ), 7 . 33 ( m , 2h ), 7 . 51 - 7 . 57 ( m , 2h ), 9 . 78 ( m , 1h ), 10 . 13 ( m , 1h ), 13 . 27 ( s , 0 . 5h ), 13 . 62 ( s , 0 . 5h ), 13 . 99 ( s , 0 . 5h ), 14 . 49 ( s , 0 . 5h ). ms m / z 549 [ m − h ] − to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( hydrazinecarbonyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 376 , 160 mg , 0 . 22 mmol ) in n - butanol ( 2 ml ) was added 5 - cyano - 2 - methyl pyridine ( 65 mg , 0 . 55 mmol ) and potassium carbonate ( 16 mg , 0 . 12 mmol ). the reaction mixture was heated to 150 ° c . under microwave irradiation for 50 minutes . the reaction was cooled and concentrated in vacuo . the residue was partitioned between water and ethyl acetate . the organic extracts were washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 15 % meoh in dcm followed by preparative tlc . the residue was treated with boron tribromide ( 0 . 047 ml 0 . 47 mmol ) and the reaction stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo and partitioned between saturated aqueous nahco 3 and etoac . the organic extracts were dried over sodium sulfate , concentrated in vacuo and purified by preparative tlc to afford the title compound ( 19 mg , 41 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 22 ( s , 3h ), 3 . 03 ( s , 3h ), 3 . 07 ( s , 3h ), 3 . 71 ( m , 2h ), 4 . 75 ( br m , 1h ), 5 . 05 ( br m , 1h ), 6 . 78 ( s , 1h ), 6 . 96 ( m , 1h ), 7 . 22 - 7 . 38 ( m , 4h ), 7 . 40 ( m , 2h ), 7 . 75 ( m , 1h ), 8 . 93 ( m , 1h ), 10 . 01 ( br m , 1h ), 13 . 80 ( br m , 1h ). ms m / z 682 [ m + h ] + to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( 5 -( piperidin - 4 - yl )- 4h - 1 , 2 , 4 - triazol - 3 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 371 , 70 mg , 0 . 097 mmol ) in dcm ( 12 ml ) was added triethylamine ( 0 . 02 ml , 0 . 146 mmol ) and trimethylsilylisocyanate ( 0 . 013 ml , 0 . 097 mmol ). the reaction was quenched with water , and the organic extracts were dried over sodium sulfate and concentrated in vacuo . the residue was dissolved in dcm ( 10 ml ) and boron tribromide ( 0 . 041 ml , 0 . 41 mmol ) was added at 0 ° c . and stirred for 3 hours . the reaction was quenched with saturated aqueous sodium bicarbonate solution followed by extraction with 20 % ipa in dcm . the organic extracts were collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc eluting with 10 % meoh in dcm to afford the title compound ( 34 mg , 78 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 42 ( m , 2h ), 1 . 75 ( m , 2h ), 2 . 67 - 2 . 75 ( m , 2h ), 3 . 01 ( s , 3h ), 3 . 08 ( s , 3h ), 3 . 17 ( m , 1h ), 3 . 89 ( m , 2h ), 4 . 34 ( m , 2h ), 4 . 69 ( m , 1h ), 4 . 99 ( m , 1h ), 5 . 93 ( br s , 2h ), 6 . 76 ( m , 1h ), 6 . 86 ( m , 1h ), 6 . 99 ( m , 1h ), 7 . 36 ( m , 1h ), 7 . 75 ( m , 1h ), 9 . 66 ( br s , 1h ), 10 . 38 - 10 . 50 ( m , 2h ), 13 . 72 ( br s , 0 . 5h ), 14 . 05 ( br s , 0 . 5h ), 14 . 20 ( br s , 0 . 5h ), 14 . 72 ( br s , 0 . 5h ). ms m / z 734 [ m + h ] + to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( 5 -( piperidin - 4 - yl )- 4h - 1 , 2 , 4 - triazol - 3 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- phenyl )- n - methylmethanesulfonamide ( preparation 372 , 90 mg , 0 . 13 mmol ) and 2 -( pyrrolidin - 1 - yl ) acetic acid ( 21 mg , 0 . 13 mmol ) in dcm ( 10 ml ) was added dipea ( 0 . 065 ml , 0 . 39 mmol ) followed by bop ( 58 mg , 0 . 13 mmol ). the reaction was allowed to stir at room temperature for 18 hours . the reaction was concentrated in vacuo and partitioned between 20 % ipa in dcm and water . the organic extracts were washed with brine , dried and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 12 % meoh in dcm . the residue was dissolved in dcm and treated with boron tribromide ( 0 . 083 ml , 0 . 87 mmol ) at 0 ° c . the reaction was stirred at room temperature for 18 hours before the addition of another aliquot of boron tribromide ( 0 . 25 ml ) and further stirring for 3 hours . the reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution and extracted with 20 % ipa in dcm . the organic extract was separated , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 15 % meoh in dcm to afford the title compound ( 41 mg , 42 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 33 ( m , 1h ), 1 . 48 ( m , 1h ), 1 . 71 - 1 . 90 ( m , 6h ), 2 . 59 - 2 . 70 ( m , 3h ), 2 . 97 ( m , 1h ), 3 . 06 ( m , 4h ), 3 . 13 ( s , 3h ), 3 . 31 ( m , 2h ), 3 . 90 ( m , 1h ), 4 . 08 ( m , 1h ), 4 . 19 ( m , 1h ), 4 . 31 ( m , 2h ), 4 . 79 ( br m , 1h ), 5 . 12 ( br m , 1h ), 6 . 99 ( m , 1h ), 7 . 35 - 7 . 59 ( m , 4h ), 7 . 73 ( m , 1h ), 10 . 44 ( m , 2h ), 13 . 93 ( br s , 1h ), 14 . 55 ( br s , 1h ). ms m / z 786 [ m + h ] + the title compound was prepared according to the method described for example 29 using n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( 4 , 5 , 6 , 7 - tetrahydro - 1h - imidazo [ 4 , 5 - c ] pyridin - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 370 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 32 - 2 . 67 ( m , 2h ), 2 . 91 ( m , 2h ), 3 . 02 ( s , 3h ), 3 . 10 ( s , 3h ), 3 . 32 ( m , 2h ), 4 . 34 ( m , 2h ), 4 . 67 ( m , 1h ), 5 . 00 ( m , 1h ), 6 . 72 ( m , 1h ), 6 . 74 ( m , 1h ), 6 . 98 ( m , 1h ), 7 . 33 ( m , 1h ), 7 . 72 ( m , 1h ), 9 . 65 ( m , 1h ), 11 . 26 - 11 . 33 ( m , 1h ), 12 . 62 ( m , 1h ), 13 . 59 ( m , 1h ). ms m / z 662 [ m + h ] + to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( 4 , 5 , 6 , 7 - tetrahydro - 1h - imidazo [ 4 , 5 - c ] pyridin - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 370 , 200 mg , 0 . 29 mmol ) and dipea ( 0 . 096 ml , 0 . 58 mmol ) in dcm ( 35 ml ) at 0 ° c . was added acetyl chloride ( 0 . 021 ml , 0 . 29 mmol ) and the reaction stirred at room temperature for 2 hours . the reaction was concentrated in vacuo and partitioned between etoac and water . the organic phase was dried , concentrated in vacuo and purified by silica gel column chromatography eluting with 5 % meoh in dcm . the residue was dissolved in dcm ( 10 ml ) and treated with boron tribromide ( 0 . 083 ml , 0 . 87 mmol ) at 0 ° c . the reaction was stirred at room temperature for 18 hours before the addition of another aliquot of boron tribromide ( 0 . 25 ml ) with further stirring for 3 hours . the reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution and extracted with 20 % ipa in dcm . the organic extract was separated , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc eluting with 10 % meoh in dcm to afford the title compound ( 25 mg , 38 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm ( tautomers ?) 2 . 50 ( m , 3h ), 2 . 62 ( m , 2h ), 3 . 02 ( s , 3h ), 3 . 15 ( s , 3h ), 3 . 70 ( m , 3h ), 4 . 35 ( m , 4h ), 4 . 54 ( m , 1h ), 5 . 00 ( m , 1h ), 6 . 71 - 7 . 00 ( m , 3h ), 7 . 35 ( m , 1h ), 7 . 70 ( m , 1h ), 9 . 60 ( m , 1h ), 10 . 37 ( m , 1h ), 11 . 54 - 11 . 67 ( m , 1h ), 13 . 65 ( s , 1h ). ms m / z 704 [ m + h ] + to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( 4 , 5 , 6 , 7 - tetrahydro - 1h - imidazo [ 4 , 5 - c ] pyridin - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl )- amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 370 , 80 mg , 0 . 116 mmol ) in dcm ( 5 ml ), was added boron tribromide ( 0 . 077 ml , 0 . 81 mmol ) at 0 ° c . and stirred for 2 hours . another aliquot of boron tribromide ( 7 eq ) was added and the reaction mixture was stirred at room temperature for a further 2 hours . the reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution and extracted with 20 % ipa in dcm . the organic extract was separated , dried over sodium sulfate and concentrated in vacuo . the residue was purified by preparative tlc . the residue was dissolved in thf ( 1 ml ) and added to a solution of n , n - dimethylamine ( 6 ml ) in thf ( 1 . 5 ml ) and bromoethylisocyanate ( 0 . 02 ml , 0 . 18 mmol ) that had stirred at 0 ° c . for 10 minutes . the reaction was stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and purified using preparative tlc to afford the title compound ( 15 mg , 21 %). 25 minute hplc qc ( sunfire c18 ( 150 × 4 . 6 mm × 5u ), mobile phase a = mecn , mobile phase b = 10 mm ammonium acetate in water rt = 2 . 59 minutes . ms m / z 776 [ m + h ] + the title compound was prepared according to the method described for example 157 using tert - butyl 2 - iodo - 6 , 7 - dihydro - 1h - imidazo [ 4 , 5 - c ] pyridine - 5 ( 4h )- carboxylate ( wo20131014567a1 ) and racemic 3 -(( 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethylsilyl ) ethoxy )- methoxy ) phenyl )- 3 - iodo - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino )- 2 - methylpropan - 1 - ol ( preparation 380 ). ms m / z 521 [ m + h ] + the title compound was prepared according to the method described for example 161 using racemic 2 - fluoro - 4 -( 4 -(( 3 - hydroxy - 2 - methylpropyl ) amino )- 3 -( 4 , 5 , 6 , 7 - tetrahydro - 1h - imidazo [ 4 , 5 - c ] pyridin - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 6 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( example 172 ). the residue was purified by preparative hplc . 10 minute hplc qc ( gemini nx - c18 ( 50 × 4 . 6 mm × 3u ), mobile phase a = 0 . 05 % formic acid in water , mobile phase b = mecn rt = 4 . 20 minutes ms m / z 611 [ m + h ] + the title compound was prepared according to the method described for preparation 11 using n -( 2 -((( 6 -( 5 - fluoro - 4 - hydroxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - hydroxyphenyl )- n - methylmethanesulfonamide ( preparation 271 ). ms m / z 585 [ m + h ] + to a solution of 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl )- n - methyl - 4 -{[ 2 -( methylamino ) benzyl ] amino }- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 14 , 500 mg , 0 . 705 mmol ) in thf ( 2 ml ) was added nah ( 28 . 2 mg , 0 . 70 mmol ) at 0 ° c . followed by dropwise addition of sulfamoyl chloride ( 97 mg , 0 . 84 mmol ). the reaction was allowed to warm to room temperature for 1 hour . the reaction was quenched with water and extracted with ethyl acetate . the organic extracts were separated , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 40 % etoac in hexanes to afford the title compound as an off - white solid ( 350 mg , 62 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), − 0 . 02 ( s , 9h ), 0 . 80 ( t , 2h ), 0 . 84 - 0 . 91 ( m , 5h ), 2 . 59 ( m , 2h ), 2 . 83 ( s , 3h ), 3 . 01 ( s , 3h ), 3 . 57 ( t , 2h ), 3 . 75 ( t , 2h ), 4 . 75 ( br s , 1h ), 5 . 00 ( br s , 1h ), 5 . 29 ( s , 2h ), 5 . 72 ( s , 2h ), 6 . 96 ( s , 1h ), 7 . 04 ( s , 1h ), 7 . 13 ( m , 1h ), 7 . 22 - 7 . 43 ( m , 4h ), 8 . 85 ( m , 1h ), 9 . 68 ( m , 1h ). ms m / z 788 [ m + h ] + to a solution of 4 -[( 2 -{[( 3 - chloropropyl ) sulfonyl ]( methyl ) amino } benzyl ) amino ]- 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 1 -{[ 2 -( tri - methylsilyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 4 , 290 mg , 0 . 32 mmol ) in etoh ( 2 ml ) was added morpholine ( 0 . 5 ml ) and the reaction was heated to 110 ° c . under microwave irradiation for 75 minutes . the reaction was cooled , concentrated in vacuo and partitioned between etoac and water . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc to afford the title compound ( 70 mg , 80 %). ms m / z 940 [ m + h ] + to a solution of 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 - fluoro - 4 -{[ 2 -( trimethyl - silyl ) ethoxy ]- methoxy } phenyl )- n - methyl - 4 -{[ 2 -( methylamino ) benzyl ] amino }- 1 -{[ 2 -( trimethylsilyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 17 , 150 mg , 0 . 19 mmol ) in thf ( 10 ml ) was added 1h - pyrazole - 4 - suffonylchloride ( 0 . 03 ml , 0 . 19 mmol ) at 0 ° c . the reaction mixture was allowed to stir at room temperature for 18 hours . the reaction was concentrated in vacuo and purified using silica gel column chromatography to afford the title compound ( 76 mg , 43 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), − 0 . 03 ( s , 9h ), 0 . 79 - 0 . 89 ( m , 4h ), 2 . 86 ( d , 3h ), 2 . 99 ( s , 3h ), 3 . 57 ( t , 2h ), 3 . 76 ( m , 4h ), 4 . 80 ( br m , 1h ), 4 . 97 ( br m , 1h ), 5 . 30 ( s , 2h ), 5 . 73 ( s , 2h ), 6 . 70 ( m , 1h ), 7 . 09 ( s , 1h ), 7 . 18 - 7 . 71 ( m , 5h ), 7 . 71 ( s , 1h ), 8 . 29 ( s , 1h ), 8 . 90 ( m , 1h ), 9 . 84 ( m , 1h ), 13 . 75 ( s , 1h ). ms m / z 893 [ m + h ] + the following preparations ( preparations 4 - 10 ) were prepared according to the method described for preparation 3 using either 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl )- ethoxy ] methoxy } phenyl ]- 4 -{[ 2 -( methylamino ) benzyl ] amino }- 1 -{[ 2 -( trimethylsilyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 16 ) or 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl )- n - methyl - 4 -{[ 2 -( methylamino ) benzyl ] amino }- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 17 ) and the appropriate sulfonyl chloride as described below : to a solution of n -( 2 -{[( 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethyls - ilyl ) ethoxy ]- methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl )- amino ] methyl }- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 85 , 250 mg , 0 . 26 mmol ) in meoh ( 4 ml ) was added molybdenum hexacarbonyl ( 84 . 91 mg , 0 . 32 mmol ), dbu ( 0 . 119 ml , 0 . 80 mmol ) and pd ( oac ) 2 ( 4 mg , 0 . 02 mmol ). the reaction was heated to 125 ° c . for 15 minutes under microwave irradiation . the reaction was cooled , diluted with etoac and filtered through celite . the filtrate was concentrated in vacuo and purified using silica gel column chromatography eluting with 10 % meoh in dcm to afford the title compound ( 100 mg , 51 %). ms m / z 858 [ m + h ] + the title compound was prepared according to the method described for preparation 11 using n - methyl - n -( 2 -{[( 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ]- methyl } phenyl ) methanesulfonamide ( preparation 105 ). ms m / z 829 [ m + h ] + to a solution of 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- n - methyl - 4 -{[ 2 -( methylamino ) benzyl ] amino }- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 17 , 50 mg , 0 . 06 mmol ) in thf ( 5 ml ) was added sodium hydride ( 1 . 88 mg , 0 . 08 mmol ) at 0 ° c . after stirring for 2 minutes , dimethylsulfamoyl chloride ( 15 mg , 0 . 11 mmol ) was added and the reaction was stirred for 1 hour . the reaction was partitioned between etoac and water , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was dissolved in dmf ( 1 ml ) and treated with cesium carbonate ( 64 mg , 0 . 19 mmol ) followed by methyl iodide ( 27 mg , 0 . 19 mmol ). the reaction was stirred at room temperature for 18 hours before quenching with ammonium chloride and extraction with etoac . the organic layer was collected and purified using preparative tlc to afford the title compound ( 45 mg , 78 %). ms m / z 870 [ m + h ] + to a solution of benzyl [ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl )- ethoxy ] methoxy } phenyl )- 3 -( methylcarbamoyl )- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino }- methyl ) phenyl ] methylcarbamate ( preparation 18 , 775 mg , 0 . 91 mmol ) in etoh ( 25 ml ) was added 10 % pd / c ( 100 mg ) and the reaction was hydrogenated at room temperature at 30 psi for 1 hour . the reaction was filtered , the filtrate was concentrated in vacuo and the residue was purified by silica gel column chromatography eluting with 15 % etoac in hexanes to afford the title compound ( 530 mg , 81 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 81 ( t , 2h ), 0 . 88 ( t , 2h ), 1 . 00 ( t , 2h ), 2 . 38 ( s , 3h ), 2 . 69 ( m , 2h ), 2 . 82 ( s , 3h ), 3 . 56 ( t , 2h ), 3 . 77 ( t , 2h ), 4 . 59 ( m , 2h ), 5 . 33 ( s , 2h ), 5 . 71 ( s , 2h ), 6 . 02 ( m , 1h ), 6 . 44 ( m , 1h ), 6 . 55 ( m , 1h ), 6 . 98 ( s , 1h ), 7 . 04 - 7 . 19 ( m , 2h ), 7 . 22 ( m , 1h ), 8 . 84 ( m , 1h ), 9 . 69 ( m , 1h ). ms m / z 709 [ m + h ] + the title compound was prepared according to the method described for preparation 14 using 6 -[ 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 4 -[({ 2 -[ methyl ( methylsulfonyl ) amino ] pyridin - 3 - yl } methyl ) amino ]- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 22 ). the residue was purified by silica gel column chromatography eluting with 25 % etoac in dcm . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 58 ( m , 2h ), 1 . 74 ( m , 1h ), 1 . 93 - 2 . 10 ( m , 3h ), 3 . 10 ( s , 1h ), 3 . 12 ( s , 3h ), 3 . 61 ( m , 1h ), 3 . 74 ( m , 2h ), 3 . 90 ( m , 1h ), 4 . 82 ( m , 2h ), 5 . 88 ( m , 1h ), 6 . 95 ( d , 1h ), 7 . 03 ( s , 1h ), 7 . 25 ( d , 1h ), 7 . 36 ( m , 1h ), 7 . 81 ( m , 1h ), 7 . 97 ( br s , 1h ), 8 . 19 ( br s , 1h ), 8 . 41 ( m , 1h ), 9 . 82 ( t , 1h ), 10 . 15 ( s , 1h ). ms m / z 652 [ m + h ] + the title compound was prepared according to the method described for preparation 14 using benzyl ( 2 -{[( 3 - carbamoyl - 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl )- ethoxy ] methoxy } phenyl ]- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl )- amino ] methyl } phenyl ) methylcarbamate ( preparation 23 ). the residue was purified by silica gel column chromatography eluting with 30 % etoac in hexanes . ms m / z 749 [ m + h ] + the title compound was prepared according to the method described for preparation 14 using benzyl [ 2 -({[ 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy }- phenyl )- 3 -( methylcarbamoyl )- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ] methylcarbamate ( preparation 19 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 81 ( t , 2h ), 0 . 89 ( t , 2h ), 2 . 50 ( s , 3h ), 2 . 84 ( d , 3h ), 3 . 58 ( t , 2h ), 3 . 76 ( t , 2h ), 3 . 95 ( q , 2h ), 4 . 58 ( d , 2h ), 5 . 33 ( s , 2h ), 5 . 73 ( s , 2h ), 5 . 74 ( br s , 1h ), 6 . 45 - 6 . 54 ( m , 2h ), 7 . 05 - 7 . 11 ( m , 3h ), 7 . 35 - 7 . 39 ( m , 2h ), 8 . 87 ( m , 1h ), 9 . 73 ( m , 1h ). ms m / z 763 [ m + h ] + to a solution of benzyl [ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ] methylcarbamate ( preparation 91 , 1 g , 1 mmol ) in methylamine / thf ( 10 ml ) was added dbu ( 0 . 49 ml , 3 . 23 mmol ), pd ( oac ) 2 ( 17 mg , 0 . 08 mmol ) followed by molybdenum hexacarbonyl ( 0 . 29 mg , 1 . 09 mmol ). the reaction was heated to 100 ° c . under microwave irradiation for 10 minutes . the reaction was cooled , concentrated in vacuo and diluted with etoac . the mixture was filtered through celite , the filtrate concentrated in vacuo and purified using silica gel column chromatography eluting with 47 % etoac in hexanes to afford the title compound ( 775 mg , 84 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 10 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 79 ( t , 2h ), 0 . 90 ( m , 5h ), 2 . 57 ( m , 2h ), 2 . 83 ( d , 3h ), 3 . 08 ( s , 1h ), 3 . 59 ( t , 2h ), 3 . 74 ( t , 2h ), 4 . 53 ( m , 1h ), 4 . 71 ( m , 1h ), 4 . 88 ( m , 1h ), 5 . 00 ( m , 1h ), 5 . 27 ( s , 1h ), 5 . 75 ( s , 2h ), 6 . 98 ( s , 1h ), 7 . 08 - 7 . 42 ( m , 11h ), 8 . 83 ( m , 1h ), 9 . 67 ( m , 1h ). ms m / z 843 [ m + h ] + a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl )- ethoxy ] methoxy } phenyl ]- n - methyl - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide - 5 - oxide ( preparation 117 , 3 . 2 g , 4 . 96 mmol ) in dmf ( 100 ml ) was treated with benzyl methyl [ 2 -({[( 4 - nitrophenoxy ) carbonyl ] amino }- methyl ) phenyl ] carbamate ( preparation 178 , 2 . 68 g , 6 . 16 mmol ) and triethylamine ( 0 . 68 ml , 4 . 96 mmol ) and heated at 80 ° c . for 16 hours . further benzyl methyl [ 2 -({[( 4 - nitrophenoxy ) carbonyl ] amino } methyl ) phenyl ] carbamate ( 1 . 24 eq ) and triethylamine ( 1 eq ) were added and the reaction allowed to continue for 6 hours . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography to afford the title compound as an oil ( 4 . 2 g , 94 %). ms m / z 897 [ m + h ] + the title compound was prepared according to the method described for preparation 18 using n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ] amino } methyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 79 ) and 6 - methylpyridin - 3 - amine with dbu at 100 ° c . for 10 minutes under microwave irradiation . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography eluting with 7 % heptanes in etoac . ms m / z 864 [ m + h ] + to a solution of n -( 2 -{[( 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl )- ethoxy ]- methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl )- amino ] methyl }- 5 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 89 , 350 mg , 0 . 37 mmol ) in meoh ( 2 ml ) was added dbu ( 0 . 16 ml , 1 . 19 mmol ), palladium acetate ( 5 . 86 mg , 0 . 03 mmol ) and molybdenum hexacarbonyl ( 99 mg , 0 . 37 mmol ) and the reaction was heated to 100 ° c . for 10 minutes under microwave irradiation . the reaction was cooled , concentrated in vacuo and purified directly using silica gel column chromatography eluting with 12 % meoh in dcm . the resulting residue was dissolved in anhydrous thf ( 5 ml ) and nmm ( 0 . 033 ml , 0 . 30 mmol ) was added followed by isobutylchloroformate ( 0 . 04 ml , 0 . 30 mmol ) at − 20 ° c . the reaction was stirred for 2 hours at this temperature before the addition of aqueous ammonia ( 0 . 5 ml ) with further stirring for 1 hour . the reaction was partitioned between etoac and water , the organic layer collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 42 % etoac in hexanes to afford the title compound ( 102 mg , 32 % over 2 steps ). ms m / z 857 [ m + h ] + the following preparations ( preparations 22 - 26 ) were prepared according to the method described for preparation 21 using the appropriate iodo intermediate as described below : to a solution of n -( 2 -((( 6 -( 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ) methyl ) phenyl )- n - methyl - methanesulfonamide ( preparation 60 , 5 . 8 g , 7 . 04 mmol ) in thf ( 15 ml ) was added molybdenum hexacarbonyl ( 1 . 872 g , 7 . 04 mmol ), dbu ( 3 . 15 ml ) and pd ( oac ) 2 ( 111 mg , 0 . 15 mmol ), and t - butyl amine ( 6 ml ). the reaction was heated in a sealed tube to 100 ° c . for 45 minutes . the reaction was cooled , filtered and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 29 % etoac in hexanes to afford the title compound ( 4 g , 71 %). 1 h nmr ( 400 mhz , dmso - d 6 ): 5 ppm 1 . 45 ( s , 9h ), 1 . 67 - 1 . 73 ( m , 2h ), 1 . 91 - 2 . 02 ( m , 2h ), 2 . 44 ( m , 2h ), 3 . 05 ( s , 3h ), 3 . 10 ( s , 3h ), 3 . 66 - 3 . 95 ( m , 4h ), 4 . 78 - 4 . 86 ( m , 2h ), 5 . 21 ( s , 2h ), 5 . 86 ( m , 1h ), 7 . 04 ( s , 1h ), 7 . 27 - 7 . 50 ( m , 11h ), 7 . 73 ( s , 1h ), 9 . 66 ( t , 1h ). ms m / z 797 [ m + h ] + the following preparations ( preparations 28 - 38 ) were prepared according to the method described for preparation 27 using the appropriate iodo intermediate as described below : to a solution of 4 - chloro - 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- n - methyl - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 118 , 150 mg , 0 . 27 mmol ) in n - butanol ( 4 ml ) was added n -[ 3 -( aminomethyl ) pyrazin - 2 - yl ]- n - methylbenzenesulfonamide ( preparation 219 , 114 mg , 0 . 41 mmol ) and dipea ( 0 . 17 ml 0 . 96 mmol ). the reaction was heated to 90 ° c . in a sealed tube for 18 hours . the reaction was quenched by the addition of water and extracted with etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 30 - 50 % etoac in hexanes to afford the title compound as a yellow solid ( 110 mg , 51 %). ms m / z 789 [ m + h ] + the following preparations ( preparations 40 - 59 ) were prepared according to the method described for preparation 39 using the appropriate chloropyridine and the appropriate amine as described below : to a solution of 6 -[ 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 112 , 110 mg , 0 . 18 mmol ) in dmf ( 5 ml ) was added 4 - nitrophenyl { 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } carbamate ( preparation 166 , 82 mg , 0 . 22 mmol ) followed by triethylamine ( 0 . 06 ml , 0 . 438 mmol ). the reaction was heated to 100 ° c . for 16 hours . further 4 - nitrophenyl { 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } carbamate ( 1 . 2 eq ) and triethylamine ( 2 . 5 eq ) were added and the reaction continued at 100 ° c . for 18 hours . the reaction was cooled , concentrated in vacuo and partitioned between ice - water and etoac . the organic layer was collected , washed with saturated aqueous potassium carbonate solution , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 49 % etoac in hexanes to afford the title compound ( 120 mg , 83 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 50 - 1 . 75 ( m , 3h ), 1 . 89 ( m , 2h ), 2 . 32 ( m , 1h ), 3 . 05 ( s , 3h ), 3 . 06 ( s , 3h ), 3 . 53 ( m , 1h ), 3 . 70 - 3 . 73 ( m , 2h ), 3 . 87 ( m , 1h ), 4 . 74 ( m , 1h ), 4 . 92 ( m , 1h ), 5 . 21 ( s , 2h ), 5 . 78 ( m , 1h ), 6 . 82 ( t , 1h ), 7 . 11 ( s , 1h ), 7 . 27 - 7 . 50 ( m , 11h ). ms m / z 824 [ m + h ] + a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 114 , 650 mg , 0 . 91 mmol ) in dmf was treated with 4 - nitrophenyl { 5 - chloro - 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } carbamate ( preparation 156 , 564 . 76 mg , 1 . 36 mmol ) and triethylamine ( 0 . 31 ml , 2 . 27 mmol ) and the reaction was heated to 90 ° c . for 16 hours . further 4 - nitrophenyl { 5 - chloro - 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } carbamate ( 1 . 5 eq ) and tea ( 1 . 5 eq ) were added and the reaction was heated to 90 ° c . for a further 4 hours . the reaction was cooled and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 30 % etoac in hexanes to afford the title compound ( 365 mg , 42 %). ms m / z 944 [ m 35 cl + h ] + the following preparations ( preparations 62 - 105 ) were prepared according to the method described for preparation 61 using 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 114 ) or 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 113 ) or 6 -[ 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 112 ) and the appropriate aminocarbamate . the following preparations ( preparations 106 - 109 ) were prepared according to the method described for preparation 61 using 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 111 ) or 6 -( 2 - cyclopropyl - 5 - fluoro - 4 - methoxyphenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 115 ) and the appropriate aminocarbamate . the title compound was prepared according to the method described for preparation 61 using 6 -( 2 - cyclopropyl - 5 - fluoro - 4 - methoxyphenyl )- n - methyl - 1 -{[ 2 -( trimethylsilyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide 5 - oxide ( preparation 116 ) and 4 - nitrophenyl ({ 2 -[ methyl ( methylsulfonyl ) amino ] pyridin - 3 - yl } methyl ) carbamate ( preparation 177 ). ms m / z 684 [ m + h ] + to a stirred solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 131 , 5 . 50 g , 9 . 45 mmol ) in dry dcm ( 550 ml ) at 0 ° c ., was added mcpba ( 1 . 79 g , 10 . 40 mmol ) portionwise followed by stirring at room temperature for 16 hours . the reaction was quenched with saturated aqueous sodium bicarbonate solution and saturated aqueous sodium bisulphite solution , the organic extracts separated , dried and purified by silica gel column chromatography eluting with etoac to afford the title compound ( 3 . 40 g , 50 %). 1 h nmr ( 400 mhz , dmso - ds ): δ ppm − 0 . 12 ( s , 9h ), 0 . 783 ( m , 2h ), 3 . 46 - 3 . 75 ( m , 4h ), 3 . 92 ( s , 3h ), 5 . 77 ( m , 2h ), 7 . 25 ( d , 1h ), 7 . 36 ( d , 1h ), 8 . 13 ( s , 1h ), 8 . 63 ( s , 1h ). ms m / z 598 [ m + h ] + to a stirred solution of 6 -[ 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 132 , 17 . 7 g , 29 mmol ) in anhydrous dcm ( 900 ml ) at 0 ° c . was added mcpba ( 7 . 51 g , 43 . 5 mmol ) and the reaction was stirred warming to room temperature for 18 hours . the reaction was quenched by the addition of saturated sodium sulphite solution ( 600 ml ) followed by saturated aqueous sodium bicarbonate solution ( 600 ml ). the organic layer was collected , washed with water ( 3 × 50 ml ), brine ( 2 × 50 ml ), dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 56 - 80 % etoac in hexanes to afford the title compound ( 13 g , 71 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 55 - 1 . 67 ( m , 3h ), 1 . 98 ( m , 2h ), 2 . 33 ( m , 1h ), 3 . 51 ( m , 1h ), 3 . 68 ( m , 2h ), 3 . 88 ( m , 1h ), 5 . 28 ( s , 2h ), 5 . 90 ( m , 1h ), 7 . 30 - 7 . 51 ( m , 7h ), 8 . 60 ( s , 1h ), 8 . 80 ( s , 1h ). the following preparations ( preparations 113 - 117 ) were prepared according to the method described for preparation 111 using the appropriate pyrrolopyridine as described below : to a solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- n - methyl - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 125 , 2 g , 3 . 90 mmol ) in anhydrous dcm ( 30 ml ) was added mcpba ( 1 . 2 g , 4 . 29 mmol ) at 0 ° c . and the reaction was stirred at room temperature for 18 hours . the reaction was quenched by the addition of saturated aqueous sodium bisulfite and sodium bicarbonate solutions and extracted into dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 15 % meoh in dcm to afford the intermediate n - oxide . this intermediate was dissolved in dmf ( 20 ml ) and oxalyl chloride ( 2 . 43 ml , 28 . 38 mmol ) was added at 0 ° c . with stirring for 1 hour . the reaction was quenched by the addition of water and extracted into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 17 % etoac in dcm to afford the title compound ( 400 mg , 26 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), 0 . 82 ( m , 2h ), 2 . 85 ( d , 3h ), 3 . 57 ( t , 2h ), 3 . 92 ( s , 3h ), 4 . 10 ( q , 2h ), 5 . 85 ( s , 2h ), 7 . 37 ( d , 1h ), 7 . 49 ( d , 1h ), 8 . 11 ( s , 1h ), 8 . 69 ( m , 1h ). ms m / z 547 [ m 35 cl + h ] + the title compound was prepared according to the method described for preparation 118 using n - ethyl - 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 129 ). the title compound may be prepared according to the method described for preparation 118 , step 1 using n - tert - butyl - 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 127 ). the n - oxide intermediate ( 1 . 3 g , 1 . 94 mmol ) was dissolved in dcm ( 150 ml ) with triethylamine ( 0 . 35 ml , 2 . 52 mmol ) and pocl 3 ( 0 . 23 ml , 2 . 52 mmol ) was added at 0 ° c . the reaction was stirred for 1 hour at 10 ° c . before quenching with saturated aqueous sodium bicarbonate solution and extraction with dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 15 % etoac in dcm to afford a yellow oil ( 530 mg , 39 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 08 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 84 ( t , 2h ), 0 . 90 ( t , 2h ), 1 . 41 ( s , 9h ), 3 . 58 ( t , 2h ), 3 . 77 ( t , 2h ), 4 . 07 ( m , 2h ), 5 . 37 ( s , 2h ), 5 . 84 ( s , 2h ), 7 . 46 ( m , 2h ), 8 . 08 ( s , 1h ), 8 . 33 ( s , 1h ). ms m / z 705 [ m + h ] + the title compound may be prepared according to the method described for preparation 118 using n -( 2 , 4 - dimethoxybenzyl )- 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 1 -{[ 2 -( trimethylsilyl )- ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 126 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 10 ( s , 9h ), 0 . 85 ( t , 2h ), 3 . 58 ( t , 2h ), 3 . 75 ( s , 3h ), 3 . 81 ( s , 3h ), 3 . 92 ( s , 3h ), 4 . 09 ( q , 2h ), 4 . 43 ( m , 2h ), 5 . 85 ( s , 2h ), 6 . 50 ( m , 1h ), 6 . 58 ( s , 1h ), 7 . 24 ( d , 1h ), 7 . 35 ( d , 1h ), 7 . 49 ( d , 1h ), 8 . 11 ( s , 1h ), 8 . 97 ( m , 1h ). ms m / z 683 [ m + h ] + the title compound may be prepared according to the method described for preparation 118 using 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- n - methyl - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 130 ). the n - oxide intermediate ( 800 mg , 1 . 24 mmol ) was dissolved in dcm ( 7 ml ) and a solution of pocl 3 ( 0 . 148 ml , 1 . 6 mmol ) in dcm ( 3 ml ) was added dropwise at 0 ° c . the reaction was stirred for 30 minutes before the addition of water and extraction into dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 14 % etoac in dcm to afford the title compound as a yellow solid ( 600 mg , 73 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 82 ( t , 2h ), 0 . 90 ( t , 3h ), 2 . 85 ( d , 3h ), 3 . 57 ( t , 2h ), 3 . 77 ( t , 2h ), 5 . 37 ( s , 2h ), 5 . 85 ( s , 2h ), 7 . 43 - 7 . 51 ( m , 2h ), 8 . 11 ( s , 1h ), 8 . 67 ( m , 1h ). ms m / z 629 [ m + h ] + to a solution of 6 -( 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- n -( tert - butyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxamide ( preparation 124 , 3 . 80 g , 6 . 5 mmol ) in anhydrous dcm ( 250 ml ) was added mcpba ( 1 . 68 g , 9 . 75 mol ) at 0 ° c . and the reaction was stirred at room temperature for 18 hours . the reaction was quenched by the addition of saturated aqueous sodium sulphite solution and saturated aqueous sodium bicarbonate solution and extracted into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with etoac to afford the intermediate n - oxide , that was dissolved in dcm ( 300 ml ). to the solution was added triethylamine ( 1 . 07 ml , 7 . 74 mmol ) followed by pocl 3 ( 0 . 62 ml , 6 . 71 mmol ) at 0 ° c . the reaction was stirred at 10 ° c . for 1 hour before the addition of ice - water . the reaction was extracted into dcm , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo to afford the title compound as the desired chloro isomer confirmed by noe irradiation of the remaining pyridyl proton . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 41 ( s , 9h ), 1 . 56 - 1 . 80 ( m , 3h ), 1 . 95 - 2 . 04 ( m , 2h ), 2 . 37 ( m , 1h ), 3 . 76 ( m , 1h ), 3 . 91 - 4 . 18 ( m , 3h ), 5 . 27 ( s , 2h ), 6 . 01 ( m , 1h ), 7 . 34 - 7 . 55 ( m , 7h ), 8 . 03 ( s , 1h ), 8 . 38 ( br s , 1h ). ms m / z 619 [ m + h ] + to a solution of 6 -[ 4 -( benzyloxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 132 , 8 g , 13 mmol ) in thf ( 30 ml ) and tert - butylamine ( 16 ml ) was added molybdenum hexacarbonyl ( 3 . 48 g , 13 mmol ), dbu ( 5 . 86 ml , 39 . 25 mmol ) and pd ( oac ) 2 ( 180 mg , 1 . 3 mmol ). the reaction was heated in a sealed tube at 100 ° c . for 1 hour . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography to afford the title compound . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 46 ( s , 9h ), 1 . 60 - 1 . 74 ( m , 4h ), 1 . 97 - 2 . 01 ( m , 2h ), 3 . 76 - 3 . 82 ( m , 1h ), 3 . 94 - 4 . 22 ( m , 3h ), 5 . 27 ( s , 2h ), 6 . 03 ( m , 1h ), 7 . 36 - 7 . 52 ( m , 7h ), 7 . 60 ( m , 1h ), 8 . 01 ( m , 1h ), 9 . 41 ( s , 1h ). ms m / z 585 [ m + h ] + to a solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -{([ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 131 , 3 . 4 g , 5 . 85 mmol ) and 2m methylamine in thf ( 30 ml ) was added palladium acetate ( 92 mg , 0 . 41 mmol ), dbu ( 2 . 62 ml , 17 . 54 mmol ) and molybdenum hexacarbonyl ( 1 . 55 g , 5 . 85 mmol ). the reaction was heated in a sealed tube at 100 ° c . for 60 minutes before concentrating in vacuo . the residue was diluted with etoac , filtered through celite and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 30 % etoac in hexanes to afford the title compound as a yellow solid ( 2 g , 67 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 15 ( s , 9h ), 0 . 80 ( t , 2h ), 2 . 85 ( d , 3h ), 3 . 56 ( t , 2h ), 3 . 92 ( s , 3h ), 4 . 18 ( q , 2h ), 5 . 87 ( s , 2h ), 7 . 33 ( d , 1h ), 7 . 41 ( d , 1h ), 8 . 07 ( s , 1h ), 8 . 66 ( m , 1h ), 9 . 45 ( s , 1h ). ms m / z 513 [ m + h ] + the following preparations ( preparations 126 - 129 ) were prepared according to the method described for preparation 125 using the appropriate pyrrolopyridine and amine as described below : the title compound may be prepared according to the method described for preparation 18 using 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ]- methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 135 ) with methylamine . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 15 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 80 ( t , 2h ), 0 . 91 ( t , 2h ), 2 . 85 ( d , 3h ), 3 . 58 ( t , 2h ), 3 . 78 ( t , 2h ), 4 . 10 ( q , 2h ), 5 . 36 ( s , 2h ), 5 . 87 ( s , 2h ), 7 . 43 ( m , 2h ), 8 . 08 ( s , 1h ), 8 . 64 ( m , 1h ), 9 . 46 ( s , 1h ). ms m / z 629 [ m + h ] + to a suspension of nah ( 0 . 59 g , 24 . 93 mmol ) in dry dmf ( 100 ml ) at 0 ° c . was added a solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 137 , 7 . 50 g , 16 . 62 mmol ) in dmf ( 100 ml ). the reaction was stirred for 30 minutes before the addition of sem - chloride ( 4 . 42 ml , 24 . 93 mmol ) drop - wise . the reaction was stirred for 1 hour before being quenched with ice - water and extracted into etoac . the organic layer was washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the crude residue was purified using silica gel column chromatography to afford the title compound as a yellow liquid ( 5 . 50 g , 47 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 15 ( s , 9h ), 0 . 76 ( m , 2h ), 3 . 51 ( m , 2h ), 3 . 91 ( s , 3h ), 4 . 11 ( q , 2h ), 5 . 81 ( s , 2h ), 7 . 33 ( d , 1h ), 7 . 42 ( d , 1h ), 7 . 99 ( s , 1h ), 8 . 85 ( s , 1h ). ms m / z 582 [ m + h ] + to a solution of 2 - fluoro - 4 -[ 3 - iodo - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 6 - yl ]- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( preparation 133 , 21 . 8 g , 41 . 8 mmol ) in acetone ( 200 ml ) was treated with benzyl bromide ( 7 . 5 ml , 62 . 7 mmol ) and potassium carbonate ( 14 . 4 g , 104 mmol ) and the reaction was heated to reflux . the reaction was cooled , filtered , the filtrate collected and concentrated in vacuo . the residue was taken up in etoac , washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 18 % etoac in hexane to afford the title compound ( 23 g , 90 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 57 - 1 . 71 ( m , 3h ), 2 . 01 ( m , 2h ), 2 . 37 ( m , 1h ), 3 . 74 ( m , 1h ), 3 . 91 ( m , 1h ), 4 . 03 ( m , 1h ), 4 . 16 ( m , 1h ), 5 . 27 ( s , 2h ), 5 . 97 ( m , 1h ), 7 . 34 - 7 . 51 ( m , 7h ), 7 . 95 ( s , 1h ), 8 . 83 ( s , 1h ). ms m / z 612 [ m + h ] + to a solution of 2 - fluoro - 4 -( 3 - iodo - 1h - pyrazolo [ 4 , 3 - c ] pyridin - 6 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( preparation 134 , 40 . 8 g , 93 mmol ) in dmf ( 500 ml ) was added dihydropyran ( 17 ml , 187 mmol ) and ptsa ( 7 . 10 g , 37 mmol ) and the reaction was heated to 80 ° c . for 18 hours . additional dihydropyran ( 2 eq ) and ptsa ( 0 . 4 eq ) were added and the reaction continued at this temperature for a further 2 hours followed by cooling to room temperature for 18 hours . the reaction was quenched by the addition of saturated aqueous sodium bicarbonate solution dropwise , and concentrated in vacuo . the aqueous residue was extracted into etoac , washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 17 - 20 % etoac in hexanes to afford the title compound as a yellow solid ( 22 g , 45 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 44 - 1 . 70 ( m , 3h ), 2 . 02 ( m , 2h ), 2 . 37 ( m , 1h ), 3 . 75 ( m , 1h ), 3 . 88 - 4 . 06 ( m , 2h ), 4 . 15 ( m , 1h ), 5 . 97 ( m , 1h ), 7 . 10 ( d , 1h ), 7 . 40 ( d , 1h ), 7 . 91 ( s , 1h ), 8 . 81 ( s , 1h ), 10 . 31 ( s , 1h ). ms m / z 522 [ m + h ] + a solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 137 , 44 g , 97 mmol ) in dcm ( 350 ml ) was treated with boron tribromide ( 46 ml , 488 mmol ) at 0 ° c ., and the reaction was allowed to stir warming to room temperature over 5 hours . the reaction was concentrated in vacuo and treated with saturated aqueous sodium bicarbonate solution . the resulting precipitate was filtered and dried under vacuum to afford the title compound as a white solid ( 41 g , 97 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 4 . 00 ( q , 2h ), 7 . 09 ( d , 1h ), 7 . 37 ( d , 1h ), 7 . 58 ( s , 1h ), 8 . 81 ( s , 1h ), 10 . 26 ( s , 1h ), 13 . 95 ( s , 1h ). ms m / z 438 [ m + h ] + the title compound was prepared according to the method described for preparation 131 using 6 -[ 4 -{[ tert - butyl ( dimethyl ) silyl ] oxy }- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 138 ). the reaction conditions cause deprotection of the tbdms ether and subsequent re - protection with sem - chloride . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 14 ( s , 9h ), − 0 . 03 ( s , 9h ), 0 . 78 ( t , 2h ), 0 . 90 ( t , 2h ), 3 . 56 ( t , 2h ), 3 . 77 ( t , 2h ), 4 . 09 ( m , 2h ), 5 . 36 ( s , 2h ), 5 . 81 ( s , 2h ), 7 . 42 ( m , 2h ), 8 . 01 ( s , 1h ), 8 . 86 ( s , 1h ). ms m / z 698 [ m + h ] + the title compound was prepared according to the method described for preparation 131 using 6 -( 2 - cyclopropyl - 5 - fluoro - 4 - methoxyphenyl )- 3 - iodo - 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 139 ). taken on to the next step without further purification . to a solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 140 , 11 . 20 g , 34 . 43 mmol ) in dmf ( 200 ml ) at 0 ° c . was added n - iodosuccinimide ( 9 . 29 g , 41 . 32 mmol ). the reaction was stirred at room temperature for 16 hours . the reaction was diluted with ethyl acetate , washed with saturated aqueous sodium bicarbonate solution and saturated aqueous sodium thiosulfate solution . the organic extracts were separated , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 22 % etoac in hexanes to afford the title compound as a white solid ( 7 . 50 g , 48 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 91 ( s , 3h ), 4 . 11 ( q , 2h ), 7 . 32 ( d , 1h ), 7 . 46 ( d , 1h ), 7 . 62 ( s , 1h ), 8 . 83 ( s , 1h ), 13 . 90 ( br s , 1h ). ms m / z 452 [ m + h ] + the title compound was prepared according to the method described for preparation 137 using 6 -[ 4 -{[ tert - butyl ( dimethyl ) silyl ] oxy }- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 1h - pyrazolo -[ 4 , 3 - c ] pyridine ( preparation 141 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 24 ( s , 6h ), 0 . 99 ( s , 9h ), 4 . 08 ( m , 2h ), 7 . 16 ( d , 1h ), 7 . 46 ( d , 1h ), 7 . 65 ( s , 1h ), 8 . 83 ( s , 1h ), 13 . 99 ( br s , 1h ). ms m / z 552 [ m + h ] + the title compound was prepared according to the method described for preparation 137 using 6 -( 2 - cyclopropyl - 5 - fluoro - 4 - methoxyphenyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 143 ). the residue was triturated with pentane and ether . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 69 ( m , 2h ), 0 . 80 ( m , 2h ), 2 . 09 ( m , 1h ), 3 . 88 ( s , 3h ), 6 . 73 ( d , 1h ), 7 . 30 ( d , 1h ), 7 . 65 ( s , 1h ), 8 . 83 ( s , 1h ), 13 . 88 ( s , 1h ). to a solution of 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 145 , 14 . 50 g , 35 . 41 mmol ) in dioxane ( 150 ml ) was added 4m hcl in dioxane ( 60 ml ). the reaction was stirred at room temperature for 16 hours before concentrating in vacuo . the residue was partitioned between etoac and saturated aqueous sodium bicarbonate solution . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo to afford the title compound that was used directly in the next reaction ( 11 . 20 g , 97 %). ms m / z 326 [ m + h ] + to a solution of 2 - fluoro - 4 -( 1h - pyrazolo [ 4 , 3 - c ] sulfate - 6 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( preparation 142 , 13 g , 41 . 76 mmol ) and 2 , 6 lutidine ( 7 . 29 ml , 62 . 65 mmol ) in anhydrous thf ( 500 ml ) at 0 ° c . was added tbdms - triflate ( 11 . 52 ml , 50 . 12 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and partitioned between water and ethyl acetate . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 20 % etoac in hexanes to afford the title compound as a white solid ( 11 g , 62 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 24 ( s , 6h ), 0 . 99 ( s , 9h ), 4 . 09 ( m , 2h ), 7 . 17 ( d , 1h ), 7 . 45 ( d , 1h ), 7 . 63 ( s , 1h ), 8 . 33 ( s , 1h ), 9 . 15 ( s , 1h ), 13 . 57 ( br s , 1h ). 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 146 , 24 g , 45 . 66 mmol ) was dissolved in tfa ( 48 ml ) at 0 ° c . and stirred for 1 hour . the reaction was concentrated in vacuo and taken up in meoh . ethylene diamine ( 2 . 4 ml ) was added at 0 ° c . and the reaction stirred for 20 minutes . the reaction was concentrated in vacuo and partitioned between ipa : dcm ( 1 : 9 ) and water . the organic extract was dried with sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography to afford the title compound as an off - white solid ( 13 g , 91 %). 1 h nmr ( 400 m 3 hz , dmso - d 6 ): δ ppm 3 . 98 ( m , 2h ), 7 . 08 ( d , 1h ), 7 . 33 ( d , 1h ), 7 . 57 ( s , 1h ), 8 . 32 ( s , 1h ), 9 . 14 ( s , 1h ), 10 . 21 ( brs , 1h ), 13 . 52 ( brs , 1h ). to a solution of 6 -( 2 - cyclopropyl - 5 - fluoro - 4 - methoxyphenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 148 , 5 g , 13 . 61 mmol ) in meoh ( 25 ml ) was added concentrated hcl ( 3 . 5 ml ) at 0 ° c . and the reaction was stirred for 16 hours . the reaction was concentrated in vacuo and partitioned between saturated aqueous nahco 3 solution and 25 % ipa in dcm . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo to afford the title compound as a white solid ( 3 . 3 g , 85 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 71 ( m , 2h ), 0 . 81 ( m , 2h ), 2 . 12 ( m , 1h ), 3 . 88 ( s , 3h ), 6 . 71 ( d , 1h ), 7 . 30 ( d , 1h ), 7 . 66 ( s , 1h ), 8 . 32 ( s , 1h ), 9 . 16 ( s , 1h ), 13 . 46 ( s , 1h ). ms m / z 284 [ m + h ] + the title compound was prepared according to the methods described by preparations 142 , 141 , 137 and 131 using 6 -[ 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 147 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 13 ( s , 9h ), − 0 . 01 ( s , 9h ), 0 . 79 ( t , 2h ), 0 . 91 ( t , 2h ), 0 . 99 ( t , 2h ), 2 . 66 ( q , 2h ), 3 . 55 ( t , 2h ), 3 . 78 ( t , 2h ), 5 . 35 ( s , 2h ), 5 . 80 ( s , 2h ), 7 . 22 ( m , 2h ), 7 . 88 ( s , 1h ), 8 . 84 ( s , 1h ). a solution of palladium acetate ( 0 . 47 g , 2 . 10 mmol ) and s - phos ( 0 . 86 g , 2 . 10 mmol ) in ethanol ( 75 ml ) was heated at 50 ° c . for 45 minutes after purging with nitrogen ( solution a ). meanwhile a solution of 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 149 , 10 g , 42 . 07 mmol ) in ethanol ( 75 ml ) was treated with 2 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolane ( wo20131014567a1 , 21 . 08 g , 63 . 10 mmol ) and an aqueous solution of potassium phosphate ( 17 . 86 g , 84 . 14 mmol ) in water ( 50 ml ) followed by purging with nitrogen for 10 minutes ( solution b ). solution a was added to solution b and the reaction heated to 80 ° c . for 18 hours before cooling and concentrating in vacuo . the residue was partitioned between ethyl acetate and water , the organic extracts collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 15 % etoac in hexanes to afford the title compound as a yellow liquid ( 14 . 10 g , 82 %). 1 h nmr ( 400 mhz , dmso - ds ): δ ppm 1 . 58 ( m , 2h ), 1 . 73 ( m , 1h ), 1 . 99 ( m , 2h ), 2 . 42 ( m , 1h ), 3 . 78 ( m , 1h ), 3 . 92 ( m , 4h ), 4 . 09 ( m , 1h ), 4 . 26 ( m , 1h ), 5 . 98 ( d , 1h ), 7 . 33 ( d , 1h ), 7 . 48 ( d , 1h ), 7 . 92 ( s , 1h ), 8 . 37 ( s , 1h ), 9 . 15 ( s , 1h ). ms m / z 410 [ m + h ] + the title compound was prepared according to the method described for preparation 145 using 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 149 ) and ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenoxy ] methoxy } ethyl )( trimethyl ) silane ( preparation 150 ). the residue was purified using silica gel column chromatography eluting with 9 % etoac in hexanes ( 19 g , 86 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 02 ( s , 9h ), 0 . 91 ( m , 2h ), 1 . 58 ( m , 2h ), 1 . 73 ( m , 1h ), 2 . 05 ( m , 2h ), 2 . 41 ( m , 1h ), 3 . 73 ( m , 2h ), 3 . 92 - 4 . 21 ( m , 2h ), 5 . 35 ( s , 2h ), 5 . 98 ( m , 1h ), 7 . 40 ( d , 1h ), 7 . 50 ( d , 1h ), 7 . 93 ( s , 1h ), 8 . 38 ( s , 1h ), 9 . 16 ( s , 1h ). the title compound was prepared according the method described for preparation 145 using ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 - ethyl - phenoxy ] methoxy } ethyl )( trimethyl ) silane ( wo20131014567a1 ) and 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 149 ). the residue was purified using silica gel column chromatography eluting with 30 % etoac in hexanes ( 34 g , 85 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 01 ( s , 9h ), 0 . 92 ( m , 2h ), 1 . 05 ( m , 3h ), 1 . 58 ( m , 2h ), 1 . 73 ( m , 1h ), 2 . 03 ( m , 2h ), 2 . 41 ( m , 1h ), 2 . 65 ( m , 2h ), 3 . 72 - 3 . 92 ( m , 4h ), 5 . 34 ( s , 2h ), 5 . 93 ( m , 1h ), 7 . 21 - 7 . 28 ( m , 2h ), 7 . 80 ( s , 1h ), 8 . 36 ( s , 1h ), 9 . 14 ( s , 1h ). ms m / z 526 [ m + h ] + the title compound was prepared according the method described for preparation 145 using 2 -( 2 - cyclopropyl - 5 - fluoro - 4 - methoxyphenyl )- 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolane ( preparation 151 ) and 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 149 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 65 - 0 . 85 ( m , 4h ), 1 . 58 ( m , 2h ), 1 . 71 ( m , 1h ), 2 . 01 ( m , 2h ), 2 . 11 ( m , 1h ), 2 . 40 ( m , 1h ), 3 . 31 ( s , 3h ), 3 . 74 ( m , 1h ), 3 . 90 ( m , 1h ), 5 . 94 ( m , 1h ), 6 . 74 ( d , 1h ), 7 . 31 ( d , 1h ), 7 . 91 ( s , 1h ), 8 . 35 ( s , 1h ), 9 . 15 ( s , 1h ). ms m / z 368 [ m + h ] + to a solution of 6 - chloro - 1h - pyrazolo [ 4 , 3 - c ] pyridine ( 75 g , 488 . 37 mmol ) in dcm ( 2 l ) was added dihydropyran ( 66 . 98 ml , 732 . 56 mmol ) followed by para - toluenesulfonic acid ( 18 . 58 g , 97 . 67 mmol ) and the reaction was heated to reflux for 18 hours . further para - toluenesulfonic acid ( 0 . 1 eq ) and dihydropyran ( 0 . 75 eq ) were added and the reaction continued heating at reflux for 6 hours . the reaction was cooled and quenched with saturated aqueous sodium bicarbonate solution . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 17 % etoac in hexanes followed by trituration with ether to afford the title compound as a pale yellow solid ( 83 g , 72 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 59 ( m , 2h ), 1 . 71 ( m , 1h ), 2 . 02 ( m , 2h ), 2 . 29 ( m , 1h ), 3 . 74 ( m , 1h ), 3 . 89 ( m , 1h ), 5 . 91 ( m , 1h ), 7 . 93 ( s , 1h ), 8 . 38 ( s , 1h ), 8 . 94 ( s , 1h ). to a solution of ( 2 -{[ 4 - bromo - 2 - fluoro - 5 -( 2 , 2 , 2 - trifluoroethyl ) phenoxy ] methoxy } ethyl )-( trimethyl ) silane ( preparation 152 , 34 g , 84 . 31 mmol ), in dry 1 , 4 - dioxane ( 1 l ) was added bis ( pinacolonato ) diboron ( 21 . 41 g , 84 . 31 mmol ) followed by koac ( 24 . 82 g , 252 . 95 mmol ). the reaction mixture was purged with nitrogen for 20 minutes before the addition of pd ( dppf ) 2 cl 2 ( 6 . 886 g , 8 . 432 mmol ) followed by further degassing for 20 minutes . the reaction was heated to reflux for 18 hours before cooling to room temperature and concentrating in vacuo . the residue was suspended in etoac and filtered through a bed of celite . the filtrate was washed with water , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 10 % etoac in hexanes to afford the title compound as an oil ( 31 g , 82 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 05 ( s , 9h ), 0 . 87 ( m , 2h ), 1 . 32 ( s , 12h ), 3 . 74 ( m , 2h ), 3 . 92 ( m , 2h ), 5 . 32 ( s , 2h ), 7 . 29 ( d , 1h ), 7 . 42 ( d , 1h ). the title compound was prepared according to the method described for preparation 150 using 1 - bromo - 2 - cyclopropyl - 5 - fluoro - 4 - methoxybenzene ( preparation 154 ). taken on to the next step as is . to a solution of 4 - bromo - 2 - fluoro - 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( preparation 153 , 25 g , 91 . 57 mmol ) in dcm ( 200 ml ) was added dipea ( 17 . 54 ml , 100 . 73 mmol ) at room temperature followed by sem - cl ( 17 . 86 ml , 100 mmol ) drop - wise at 0 ° c . and the reaction was stirred at room temperature for 4 hours . the reaction was partitioned between dcm and water , the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo to afford the title compound as an oil ( 34 g , 92 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 03 ( s , 9h ), 0 . 87 ( m , 2h ), 3 . 70 - 3 . 79 ( m , 4h ), 5 . 29 ( s , 2h ), 7 . 42 ( d , 1h ), 7 . 70 ( d , 1h ). to a solution of 1 - bromo - 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) benzene ( wo20131014567a1 , 30 g , 104 . 51 mmol ) in dcm ( 800 ml ) at 0 ° c . was added boron tribromide ( 130 . 91 g , 522 mmol ) drop - wise and the reaction was stirred at room temperature for 16 hours . the reaction was quenched by the addition of saturated aqueous nahco 3 solution and extracted into dcm . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo to afford the title compound as a white solid that was taken directly on to the next step ( 22 g , 77 %). to a solution of 4 - cyclopropyl - 1 - fluoro - 2 - methoxybenzene ( preparation 155 , 8 . 7 g , 41 mmol ) in dmf ( 250 ml ) at 0 ° c . was added nbs ( 7 . 40 g , 41 mmol ) and the reaction was stirred at room temperature for 3 hours . the reaction was partitioned between etoac and brine , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with hexanes to afford the title compound as a colorless oil ( 9 . 5 g , 92 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 73 ( m , 2h ), 0 . 96 ( m , 2h ), 2 . 01 ( m , 1h ), 3 . 82 ( s , 3h ), 6 . 72 ( d , 1h ), 7 . 50 ( d , 1h ). to a solution of 5 - bromo - 2 - fluoroanisole ( 10 g , 48 . 77 mmol ) in toluene ( 100 ml ) was added water ( 10 ml ), cyclopropyl boronic acid ( 5 . 44 g , 63 mmol ), tricyclohexylphosphine ( 1 . 37 g , 4 . 87 mmol ) and potassium phosphate ( 36 . 3 g , 170 mmol ). the reaction was degassed with nitrogen before the addition of pd ( oac ) 2 ( 547 mg , 2 . 44 mmol ) followed by heating to 100 ° c . for 3 hours . the reaction was cooled and partitioned between etoac and brine . the organic layer was collected , concentrated in vacuo and purified using silica gel column chromatography to afford the title compound as a colorless oil ( 9 . 7 g , quant ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 65 ( m , 2h ), 0 . 92 ( m , 2h ), 1 . 88 ( m , 1h ), 3 . 81 ( s , 3h ), 6 . 61 ( m , 1h ), 6 . 82 ( m , 1h ), 7 . 06 ( m , 1h ). to a solution of n -[ 2 -( aminomethyl )- 4 - chlorophenyl ]- n - methylmethanesulfonamide hydrochloride ( preparation 211 , 3 . 20 g , 11 . 39 mmol ) and sodium carbonate ( 3 . 62 g , 34 . 18 mmol ) in dcm ( 50 ml ) at 0 ° c ., was added 4 - nitrophenylchloroformate ( 2 . 52 g , 12 . 53 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and the residue purified by silica gel column chromatography eluting with 55 - 100 % etoac in hexanes to afford the title compound as a pale yellow solid ( 2 . 20 g , 46 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 08 ( s , 3h ), 3 . 17 ( s , 3h ), 4 . 32 ( br m , 1h ), 4 . 52 ( br m , 1h ), 7 . 40 - 7 . 61 ( m , 5h ), 8 . 25 ( m , 2h ). the following preparations ( preparations 157 - 183 ) were prepared according to the method described for preparation 156 using the appropriate amine as described below , and taken directly on to the next step ; the following preparations ( preparations 184 - 207 ) were prepared according to the methods described in the three steps below : 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 06 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 26 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 06 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 06 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 26 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 34 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 10 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 05 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 37 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 99 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 11 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 96 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 36 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 4 . 18 ( br m , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 98 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 99 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 98 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 98 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 00 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 95 ( t , to a suspension of tert - butyl ( 2 -(( 2 - oxooxazolidine )- 3 - sulfonamido ) benzyl ) carbamate ( preparation 251 , 2 . 5 g , 6 . 73 mmol ) and anhydrous potassium carbonate ( 2 . 32 g , 16 . 82 mmol ) in acetone ( 300 ml ) was added methyl iodide ( 2 . 39 g , 16 . 83 mmol ) and the reaction heated to reflux for 18 hours . the reaction was cooled and concentrated in vacuo . the residue was partitioned between water and dcm , the organic layer was collected , washed with brine and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 35 % etoac in hexanes before being treated with 4m hcl in dioxane ( 7 ml ) and stirring at room temperature for 18 hours . the reaction was concentrated in vacuo and triturated with ether / pentane to afford the title compound as the hydrochloride salt ( 1 . 60 g , 68 % over 2 steps ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 36 ( s , 3h ), 3 . 56 ( m , 1h ), 3 . 98 ( m , 1h ), 4 . 08 ( m , 1h ), 4 . 19 ( m , 1h ), 4 . 44 ( m , 2h ), 7 . 52 - 7 . 68 ( m , 4h ), 8 . 36 ( br s , 3h ). ms m / z 285 [ m + h ] + to a solution of n -( 2 - acetylphenyl )- n - methylmethanesulfonamide ( preparation 246 , 10 g , 43 . 99 mmol ) in etoh ( 150 ml ) was added triethylamine ( 7 . 93 ml , 57 mmol ) and hydroxylamine hydrochloride ( 3 . 98 g , 57 mmol ) and the reaction was heated to 80 ° c . for 18 hours . the reaction was cooled , diluted with etoac , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was dissolved in meoh ( 50 ml ) and ammonium formate ( 2 . 15 g , 34 mmol ) and activated zinc dust ( 2 . 25 g , 34 mmol ) were added . the reaction was heated to reflux for 18 hours . the reaction was filtered through celite and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 12 % meoh in dcm to afford the title compound as a colorless oil ( 1 . 2 g , 77 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 48 ( d , 3h ), 3 . 16 ( s , 3h ), 3 . 22 ( s , 3h ), 4 . 76 ( m , 1h ), 7 . 40 - 7 . 80 ( m , 4h ), 8 . 32 ( s , 1h ). ms m / z 229 [ m + h ] + to a solution of 2 -( 2 - aminoethyl ) aniline ( 30 g , 220 mmol ) in dcm ( 700 ml ) at 0 ° c . was added triethylamine ( 36 . 8 ml , 264 mmol ) followed by tert - butyldicarbonate ( 52 . 9 g , 242 mmol ) and the reaction was allowed to warm to room temperature stirring for 2 hours . the reaction was added to water ( 500 ml ), the organic layer collected , washed with brine , dried over sodium sulfate and concentrated in vacuo to afford a yellow oil . the oil was dissolved in dcm ( 400 ml ) and pyridine ( 20 ml ), and benzenesulfonyl chloride ( 26 . 1 ml , 203 mmol ) was added . the reaction was stirred at room temperature for 48 hours . further benzenesulfonyl chloride ( 6 . 51 ml , 0 . 3 eq ) was added and the reaction continued for 24 hours . the reaction was washed with 1m aqueous hcl solution ( 500 ml ), concentrated aqueous ammonia solution ( 400 ml ), brine ( 500 ml ), dried over sodium sulfate and concentrated in vacuo . the residue was recrystallised from etoac / ether to afford a white solid . the solid was dissolved in dioxane ( 200 ml ), 4m hcl in dioxane ( 282 ml ) was added and the reaction was stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and the residue suspended in hot meoh ( 150 ml ). 7m ammonia in meoh ( 150 ml ) was added and the solution cooled . the resulting precipitate was collected and purified further using silica gel column chromatography eluting with 0 . 4 % nh 3 in 10 - 15 % meoh in dcm to afford the title compound as a yellow solid ( 16 . 8 g , 26 % over three steps ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 64 ( t , 2h ), 2 . 82 ( t , 2h ), 6 . 95 - 7 . 05 ( m , 4h ), 7 . 50 - 7 . 60 ( m , 3h ), 7 . 53 ( m , 2h ). ms m / z 275 [ m − h ] − to a solution of tert - butyl { 5 - chloro - 2 -[ methyl ( methylsulfonyl )- amino ] benzyl } carbamate ( preparation 213 , 8 . 2 g , 23 mmol ) in meoh ( 100 ml ) was added 4m hcl in dioxane ( 100 ml ) at 0 ° c . and the reaction was stirred at room temperature for 5 hours . the reaction was concentrated in vacuo and triturated with a 1 : 1 mixture of mecn : ether to afford the title compound as the hydrochloride salt ( 8 . 00 g , 100 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 07 ( s , 3h ), 3 . 20 ( s , 3h ), 4 . 05 ( br m , 1h ), 4 . 20 ( br m , 1h ), 7 . 58 ( m , 1h ), 7 . 66 ( m , 1h ), 7 . 73 ( m , 1h ), 8 . 32 ( br s , 3h ). ms m / z 249 [ m + h ] the title compound was prepared according to the method described for preparation 211 using tert - butyl { 5 - methoxy - 2 -[ methyl ( pyridin - 3 - ylsulfonyl ) amino ] benzyl } carbamate ( preparation 214 ). ms m / z 308 [ m + h ] + to a solution of n -( 4 - chloro - 2 - cyanophenyl )- n - methylmethanesulfonamide ( preparation 236 , 6 . 20 g , 25 . 40 mmol ) in meoh ( 150 ml ) was added di - tert - butyl dicarbonate ( 11 . 71 ml , 50 . 82 mmol ) and nicl 2 . 6h 2 o ( 1 . 20 g , 5 . 08 mmol ). the reaction was cooled to 0 ° c . and nabh 4 ( 9 . 61 g , 254 mmol ) was added portion - wise . the reaction was stirred at room temperature for 6 hours before being quenched by the addition of diethylenetriamine with stirring for 30 minutes . the reaction was concentrated in vacuo and partitioned between etoac and saturated aqueous sodium bicarbonate . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography to afford the title compound ( 8 . 20 g , 93 %). 1h nmr ( 400 mhz , dmso - d 6 ): 3 . 06 ( s , 3h ), 3 . 21 ( s , 3h ), 4 . 01 - 4 . 19 ( m , 2h ), 7 . 54 - 7 . 57 ( q , 1h ), 7 . 65 - 7 . 68 ( d , 1h ), 7 . 82 - 7 . 83 ( d , 1h ) ms m / z 349 [ m + h ] + and 249 [ m - boc + h ] + to a solution of tert - butyl [ 5 - methoxy - 2 -( methylamino ) benzyl ] carbamate ( preparation 215 , 3 . 8 g , 10 mmol ) in thf ( 20 ml ) was added nah ( 373 mg , 15 mmol ) at 0 ° c . and the reaction was stirred for 15 minutes before the addition of pyridine - 3 - sulfonyl chloride ( 1 . 36 ml , 11 mmol ) dropwise . the reaction was stirred at room temperature for 18 hours before being quenched with water and extracted into etoac . the organic layer was collected , washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 55 % etoac in hexanes to afford the title compound ( 3 . 3 g , 78 %). ms m / z 407 [ m − h ]− the title compound was prepared according to the method described for preparation 213 using 5 - methoxy - 2 -( methylamino ) benzonitrile ( preparation 250 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 37 ( s , 9h ), 2 . 66 ( d , 3h ), 3 . 63 ( s , 3h ), 3 . 95 ( m , 2h ), 4 . 77 ( br s , 1h ), 6 . 45 ( d , 1h ), 6 . 63 ( br s , 1h ), 6 . 69 ( dd , 1h ), 7 . 23 ( br t , 1h ). ms m / z 267 [ m + h ] + to a solution of n -( 2 - cyanophenyl )- n -[ 2 -( tetrahydro - 2h - pyran - 2 - yloxy ) ethyl ] methanesulfonamide ( preparation 222 , 8 g , 25 mmol ) in meoh ( 100 ml ) was added nicl 2 . 6h 2 o ( 1 . 17 g , 5 mmol ) followed by sodium borohydride ( 6 . 53 g , 172 mmol ) at 0 ° c . the reaction was stirred at room temperature for 4 hours before being quenched by the addition of diethylenetetramine . the reaction was concentrated in vacuo and purified using silica gel column chromatography eluting with 10 % meoh in dcm to afford the title compound ( 4 . 9 g , 60 %). taken on directly to the next step . to a solution of n -( 3 - cyanopyridin - 2 - yl )- n - methylmethanesulfonamide ( preparation 230 , 10 g , 47 mmol ) in methanolic ammonia ( 100 ml ) was added raney nickel ( 2 g ) and the reaction was hydrogenated at 40 psi at room temperature for 18 hours . the reaction was filtered through celite , concentrated in vacuo and purified using silica gel column chromatography eluting with 10 % meoh in dcm to afford the title compound ( 7 . 5 g , 74 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 13 ( s , 6h ), 3 . 82 ( br s , 2h ), 7 . 44 ( m , 1h ), 8 . 03 ( m , 1h ), 8 . 37 ( m , 1h ). the title compound was prepared according to the method described for preparation 217 using n -( 3 - cyanopyridin - 2 - yl )- n - methylmethanesulfonamide ( preparation 234 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 94 ( t , 3h ), 3 . 06 ( s , 3h ), 3 . 63 ( q , 2h ), 3 . 86 ( br s , 2h ), 7 . 46 ( m , 1h ), 8 . 06 ( m , 1h ), 8 . 41 ( m , 1h ). ms m / z 230 [ m + h ] a solution of n -( 3 - cyanopyrazin - 2 - yl )- n - methylbenzenesulfonamide ( preparation 228 , 7 . 2 g , 32 mmol ) in acoh ( 100 ml ) was purged under nitrogen for 15 minutes followed by the addition of 10 % pd — c ( 1 . 4 g ) and hydrogenated under at 40 psi hydrogen in a parr - shaker for 18 hours . the reaction was filtered through celite , concentrated in vacuo , neutralized with 1n naoh and extracted with dcm . the organic layer was collected , dried over sodium sulfate , concentrated in vacuo and purified by silica gel column chromatography eluting with 10 % meoh in dcm to afford the title compound as a yellow solid ( 4 . 1 g , 46 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 06 ( s , 3h ), 4 . 04 ( s , 2h ), 7 . 17 ( m , 4h ), 7 . 33 ( m , 1h ), 8 . 34 ( d , 1h ), 8 . 64 ( d , 1h ). ms m / z 279 [ m + h ] + to a solution of tert - butyl { 2 -[( pyridin - 3 - ylsulfonyl ) amino ] benzyl } carbamate ( preparation 227 , 4 . 57 g , 12 mmol ) in acetone ( 100 ml ) was added potassium carbonate ( 5 . 20 g , 38 mmol ) followed by methyl iodide ( 1 . 56 ml , 25 mmol ). the reaction was heated to reflux for 2 hours . the reaction was evaporated to dryness and partitioned between water & amp ; ethyl acetate . the organic phase was washed with brine , dried over sodium sulfate , evaporated and purified by silica gel column chromatography eluting with 52 % etoac in hexanes . the residue was dissolved in meoh ( 25 ml ) and 4m hcl in dioxane ( 25 ml ) was added with stirring at room temperature for 4 hours . the reaction was concentrated in vacuo and triturated with mecn - ether to afford the title compound as the hydrochloride salt ( 4 . 2 g , 100 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 19 ( s , 3h ), 4 . 14 ( m , 1h ), 4 . 32 ( m , 1h ), 6 . 65 ( d , 1h ), 7 . 31 ( t , 1h ), 7 . 47 ( t , 1h ), 7 . 67 - 7 . 72 ( m , 2h ), 7 . 99 ( m , 1h ), 8 . 36 ( br s , 3h ), 8 . 71 ( m , 1h ), 8 . 95 ( m , 1h ), the title compound was prepared according to the method described for preparation 219 using n -( 3 - cyanopyrazin - 2 - yl )- n - ethylmethanesulfonamide ( preparation 229 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 00 ( t , 3h ), 1 . 86 ( br s , 2h ), 3 . 13 ( s , 3h ), 3 . 68 ( q , 2h ), 3 . 95 ( s , 2h ). ms m / z 231 [ m + h ] + to a suspension of n -( 2 - cyanophenyl ) methanesulfonamide ( preparation 223 , 7 g , 25 mmol ) and polymer bound triphenylphosphine ( 14 g , 53 mmol ) in anhydrous thf ( 100 ml ) was added dead ( 8 . 42 ml , 53 mmol ) followed by 2 -( tetrahydro - pyran - 2 - yloxy ) ethanol ( 7 . 82 g , 146 mmol ) drop - wise at 0 ° c . the reaction was stirred at room temperature for 5 hours before filtering through celite . the filtrate was concentrated in vacuo and purified using silica gel column chromatography eluting with 30 - 35 % etoac in hexanes to afford the title compound ( 8 g , 69 %). 1 h nmr ( 400 mhz , meod ): δ ppm 1 . 42 - 1 . 62 ( m , 6h ), 3 . 14 ( s , 3h ), 3 . 40 - 3 . 55 ( m , 2h ), 3 . 70 - 4 . 00 ( m , 5h ), 7 . 53 ( m , 1h ), 7 . 69 ( m , 1h ), 7 . 72 - 7 . 81 ( m , 2h ). to a solution of n -( 2 - cyanophenyl )- n -( methylsulfonyl ) methanesulfonamide ( preparation 238 , 300 g , 1 . 09 mol ) in thf ( 2 l ) was added 40 % aqueous sodium hydroxide ( 2 l ), benzyl triethylammonium chloride ( 24 . 91 g , 0 . 100 mol ), and iodomethane ( 81 . 68 ml , 1 . 31 mol .) the reaction was stirred at room temperature for 18 hours . the reaction was diluted with etoac and partitioned with brine . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was triturated in pentane - ether to afford the title compound ( 208 g , 90 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 3 . 11 ( s , 3h ), 3 . 38 ( s , 3h ), 7 . 43 - 7 . 47 ( t , 1h ), 7 . 52 - 7 . 55 ( m , 1h ), 7 . 63 - 7 . 71 ( m , 2h ); ms m / z 211 [ m − h ] − to a stirred solution of 4 - methoxy - 2 -( methylamino ) benzonitrile ( preparation 226 , 11 g , 68 mmol ) in thf at − 78 ° c ., 1m lihmds in thf ( 108 . 5 ml ) was added drop wise . the solution was stirred for 30 minutes followed by the addition of methanesulfonyl chloride ( 7 . 92 ml , 102 mmol ). the reaction was stirred for 1 hour before quenching with saturated aqueous ammonium chloride solution and extracting into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 40 % etoac in hexanes to afford the title compound as a white solid ( 12 g , 73 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 13 ( s , 3h ), 3 . 26 ( s , 3h ), 3 . 88 ( s , 3h ), 7 . 11 ( dd , 1h ), 7 . 28 ( d , 1h ), 7 . 83 ( d , 1h ). the title compound was prepared according to the method described for preparation 224 using 2 -( methylamino ) benzonitrile and 3 - methoxybenzenesulfonyl chloride . taken on directly to the next step . ms m / z 303 [ m + h ] + to a solution of 2 - fluoro - 4 - methoxybenzonitrile ( 1 g , 6 . 61 mmol ) in mecn ( 10 ml ) was added 40 % aqueous methylamine ( 20 ml ) and the reaction heated to 60 ° c . in a sealed tube . the reaction was cooled , concentrated in vacuo and purified using silica gel column chromatography eluting with 60 % etoac in hexanes to afford the title compound as a white solid ( 600 mg , 56 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 76 ( d , 3h ), 3 . 78 ( s , 3h ), 6 . 12 ( m , 2h ), 6 . 22 ( m , 1h ), 7 . 37 ( m , 1h ). ms m / z 163 [ m + h ] + to a solution of ( 2 - amino - benzyl )- carbamic acid tert butyl ester ( 3 . 2 g , 14 mmol ) in pyridine ( 25 ml ) was added pyridine - 3 - sulfonylchloride ( 1 . 75 ml , 14 mmol ) at 0 ° c . the reaction was stirred at room temperature for 4 hours before concentrating in vacuo . the residue was partitioned between etoac and water , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 65 % etoac in hexanes to afford the title compound ( 4 . 5 g , 87 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 38 ( s , 9h ), 4 . 07 ( m , 2h ), 6 . 80 ( d , 1h ), 7 . 13 ( m , 1h ), 7 . 21 ( m , 2h ), 7 . 32 ( m , 1h ), 7 . 62 ( m , 1h ), 8 . 05 ( m , 1h ), 8 . 78 - 8 . 83 ( m , 2h ), 9 . 92 ( s , 1h ). ms m / z 364 [ m + h ] + to a solution of 2 - chloro - 3 - cyanopyrazine ( 5 g , 35 . 94 mmol ) and cs 2 co 3 ( 16 . 27 g , 50 mmol ) in acetonitrile ( 75 ml ) was added n - methylbenzenesulfonamide ( 7 . 37 g , 43 mmol ) and the reaction heated to 80 ° c . for 3 hours . the reaction mixture was concentrated in vacuo and the residue partitioned between water and etoac . the organic layer was collected , dried , concentrated in vacuo and purified by silica gel column chromatography eluting with 50 % etoac in hexanes to afford the title compound ( 7 . 2 g , 73 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 12 ( s , 3h ), 7 . 64 ( m , 4h ), 7 . 76 ( m , 1h ), 8 . 80 ( d , 1h ), 8 . 85 ( d , 1h ). the title compound was prepared according to the method described for preparation 228 using n - ethylmethanesulfonamide . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 09 ( t , 3h ), 3 . 20 ( s , 3h ), 3 . 83 ( q , 2h ), 8 . 89 ( d , 1h ), 8 . 97 ( d , 1h ). to a solution of 2 - chloronicotinonitrile ( 10 g , 71 . 9 mmol ) in mecn ( 200 ml ) was added cesium carbonate ( 32 . 5 g , 99 mmol ) followed by n - methylmethanesulfonamide ( 9 . 42 g , 86 mmol ) and the reaction was heated to 80 ° c . for 3 hours . the reaction was cooled , concentrated in vacuo and partitioned between etoac and water . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 50 % etoac in hexanes to afford the title compound ( 12 . 9 g , 85 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 21 ( s , 3h ), 3 . 28 ( s , 3h ), 7 . 62 ( m , 1h ), 8 . 45 ( d , 1h ), 8 . 77 ( d , 1h ). ms m / z 212 [ m + h ] + the following preparations ( preparations 231 - 235 ) were prepared according to the method described for preparation 230 using the appropriate chloropyridine and sulfonamide as described below , and taken directly on to the next step : 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 17 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 21 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 05 ( t , a solution of n -( 4 - chloro - 2 - cyanophenyl )- n -( methylsulfonyl ) methanesulfonamide ( preparation 237 , 8 . 00 g , 25 . 91 mmol ) in thf ( 100 ml ) and 40 % aqueous naoh solution ( 100 ml ) was cooled to 0 ° c . benzyltriethylammonium chloride ( 0 . 59 g , 2 . 591 mmol ) and mel ( 5 . 64 ml , 90 . 68 mmol ) were added and the reaction was stirred for 18 hours . the reaction was partitioned between etoac and water and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 30 % etoac in hexanes to afford the title compound as a yellow solid ( 6 . 20 g , 97 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 14 ( s , 3h ), 3 . 26 ( s , 3h ), 7 . 78 ( d , 1h ), 7 . 88 ( dd , 1h ), 8 . 13 ( d , 1h ). to a solution of 2 - amino - 5 - chloro - benzonitrile ( 5 . 00 g , 32 . 77 mmol ) in pyridine ( 100 ml ) at 0 ° c . was added methanesulphonylchloride ( 10 . 21 ml , 131 . 07 mmol ) and the reaction stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and partitioned between 2n hcl and etoac . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was triturated with 1 : 1 acetonitrile : ether to afford the title compound ( 8 . 00 g , 79 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 61 ( s , 6h ), 7 . 90 ( d , 1h ), 7 . 98 ( dd , 1h ), 8 . 31 ( d , 1h ). the following preparations ( preparations 238 - 246 ) were prepared according to the method described for preparation 237 using the appropriate aniline as described below : 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 10 ( t , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 61 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 10 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 44 ( s , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 7 . 18 ( m , 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 57 ( s , to a solution of n -[ 2 -( aminomethyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 252 , 10 . 5 g , 49 mmol ) in acetone ( 250 ml ) was added potassium carbonate ( 13 . 59 g , 98 . 47 mmol ) and methyl iodide ( 6 . 13 ml , 98 . 47 mmol ) at 0 ° c . followed by heating to 60 ° c . for 4 hours . the reaction was cooled and concentrated in vacuo . the residue was partitioned between etoac and water , the organic layer was collected , washed with brine , dried and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 40 % etoac in hexanes to afford the title compound ( 10 g , 89 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 54 ( s , 3h ), 2 . 97 ( s , 3h ), 3 . 29 ( s , 3h ), 7 . 46 ( m , 1h ), 7 . 60 - 7 . 64 ( m , 3h ). ms m / z 228 [ m + h ] + the title compound was prepared according to the method described for preparation 247 using 3 - cyano - 4 - aminopyridine . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 92 ( s , 3h ), 7 . 31 ( d , 1h ), 7 . 99 ( d , 1h ), 8 . 66 ( s , 1h ), 13 . 14 ( br s , 1h ). ms m / z 198 [ m + h ] + to a solution of tert - butyl [ 2 -( methylamino ) benzyl ] carbamate ( wo2004 / 046107a1 , 1 . 7 g , 7 . 2 mmol ) in thf ( 25 ml ) at 0 ° c . was added nah ( 0 . 25 g , 10 . 8 mmol ) followed by cbz - chloride ( 1 . 22 g , 7 . 2 mmol ) and catalytic dmap ( 9 mg , 0 . 72 mmol ). the reaction was heated to reflux for 2 hours before cooling , quenching with water and extracting into etoac . the organic extracts were dried over sodium sulfate , concentrated in vacuo and purified by silica gel column chromatography . the residue was dissolved in meoh ( 10 ml ) and 20 % hcl in dioxane ( 10 ml ) was added with stirring at room temperature for 18 hours . the reaction was concentrated in vacuo , and triturated with pentane and ether to afford the title compound as the hydrochloride salt ( 1 . 2 g , 91 %). ms m / z 271 [ m + h ] + to a solution of 2 - amino - 5 - methoxybenzonitrile ( 10 g , 67 mmol ) in dmf ( 100 ml ) was added tbuok ( 9 . 46 g , 84 mmol ) followed by dimethyl oxalate ( 11 . 95 g , 101 mmol ) at 0 ° c . and the reaction was heated to 120 ° c . for 4 hours . the reaction was cooled , quenched by the addition of ice - water and extracted into etoac . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography to afford the title compound ( 2 . 9 g , 27 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 72 ( s , 3h ), 3 . 67 ( s , 3h ), 5 . 77 ( br s , 1h ), 6 . 67 ( d , 1h ), 7 . 06 ( d , 1h ), 7 . 11 ( dd , 1h ). to a solution of chlorosulfonyl isocyanate ( 2 g , 8 . 99 mmol ) in dry dcm ( 20 ml ) at 0 ° c . was added bromoethanol ( 0 . 60 ml , 8 . 19 mmol ) and the reaction was stirred at room temperature for 10 minutes . triethylamine ( 2 . 74 ml , 19 . 79 mmol ) in dcm was added followed by ( 2 - amino - benzyl )- carbamic acid tert - butyl ester ( 1 . 9 g , 8 . 19 mmol ) and the reaction stirred at room temperature for 18 hours . the reaction was quenched with water , extracted into dcm , the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 20 % etoac in hexanes to afford the title compound ( 2 . 5 g , 75 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 43 ( s , 9h ), 3 . 81 ( t , 2h ), 4 . 25 ( m , 2h ), 4 . 36 ( t , 2h ), 7 . 23 - 7 . 37 ( m , 5h ), 10 . 42 ( br s , 1h ). to a solution of 1 -( 2 - aminophenyl ) ethanone ( 2 g , 14 . 8 mmol ) in pyridine ( 20 ml ) at 0 ° c . was added methanesulfonyl chloride ( 4 . 6 ml , 59 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and partitioned between 2n hcl and etoac . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo to afford the title compound as a brown solid ( 1 . 6 g , 51 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 65 ( s , 3h ), 3 . 18 ( s , 3h ), 7 . 23 ( t , 1h ), 7 . 58 - 7 . 67 ( m , 2h ), 8 . 07 ( d , 1h ). ms m / z 212 [ m − h ] − to a solution of 4 -(( 2 -( n - ethylphenylsulfonamido )- 5 - hydroxybenzyl ) amino )- 6 -( 5 - fluoro - 4 - hydroxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 259 , 300 mg , 0 . 36 mmol ) in anhydrous thf ( 4 ml ) was added nmm ( 0 . 06 ml , 0 . 58 mmol ) and isobutylchloroformate ( 0 . 07 ml , 0 . 58 mmol ) at − 20 ° c . and the reaction was stirred at this temperature for 30 minutes . aqueous ammonia ( 0 . 2 ml ) was added and the reaction stirred at room temperature for 30 minutes . the reaction was diluted with water and extracted into etoac . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 40 % etoac in hexanes to afford the title compound ( 180 mg , 60 %). ms m / z 818 [ m + h ] + the title compound was prepared according to the method described for preparation 253 using 4 -(( 2 -( n - methylphenylsulfonamido )- 5 - hydroxybenzyl ) amino )- 6 -( 5 - fluoro - 4 - hydroxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 260 ). ms m / z 804 [ m + h ] + to a solution of ( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( methylcarbamoyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )( methyl ) carbamate ( preparation 256 , 80 mg , 0 . 13 mmol ) in anhydrous thf was added boron tribromide ( 0 . 08 ml , 0 . 9 mmol ) at 0 ° c . and the reaction was stirred at this temperature for 2 hours . another aliquot of boron tribromide was added ( 7eq ) and the reaction continued stirring for a further 2 hours . the reaction was concentrated in vacuo and dissolved in dcm . the solution was washed with saturated aqueous sodium bicarbonate solution , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using preparative tlc . the residue was dissolved in anhydrous thf ( 5 ml ). to this solution at 0 ° c . was added triethylamine ( 0 . 04 ml , 0 . 3 mmol ) followed by ditertbutyldicarbonate ( 0 . 05 ml , 0 . 21 mmol ) and catalytic dmap ( 1 mg , 0 . 008 mmol ). the reaction was stirred at room temperature for 1 hour before concentrating in vacuo . the residue was purified by preparative tlc to afford the title compound . ms m / z 704 [ m + h ] + to a solution of benzyl ( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( methylcarbamoyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )( methyl ) carbamate ( preparation 257 , 150 mg , 0 . 23 mmol ) in anhydrous thf ( 5 ml ) at 0 ° c . was added triethylamine ( 0 . 08 ml , 0 . 575 mmol ) followed by ditertbutyldicarbonate ( 60 mg , 0 . 27 mmol ) and a catalytic amount of dmap . the reaction was stirred at room temperature for 18 hours . the reaction was partitioned between etoac and brine , the organic layer collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 32 % etoac in hexanes . the residue was dissolved in ethanol ( 15 ml ) and was hydrogenated at 30 psi for 1 hour over 10 % palladium on carbon ( 10 mg ). the reaction was filtered through celite , concentrated in vacuo and purified using silica gel column chromatography eluting with 32 % etoac in hexanes to afford the title compound as a white solid ( 50 mg , 80 %). ms m / z 618 [ m + h ] + the title compound was prepared according to the method described for preparation 18 using benzyl ( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )( methyl ) carbamate ( preparation 284 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 82 ( s , 3h ), 3 . 17 ( s , 3h ), 3 . 88 ( s , 3h ), 4 . 26 - 4 . 39 ( m , 2h ), 4 . 65 ( m , 1h ), 4 . 79 - 4 . 90 ( m , 2h ), 5 . 03 ( m , 1h ), 7 . 09 - 7 . 42 ( m , 10h ), 7 . 70 ( m , 1h ), 8 . 83 ( t , 1h ), 9 . 90 ( t , 1h ), 14 . 07 ( s , 1h ). the following preparations ( preparations 258 - 261 ) were prepared according to the method described for preparation 11 using the appropriate pyrazolopyrimidine as described below . to a suspension of nah ( 0 . 163 g , 6 . 79 mmol ) in dry dmf ( 50 ml ) was added n -( 2 -((( 6 -( 4 -(( tert - butyldimethylsilyl ) oxy )- 2 - ethyl - 5 - fluorophenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylbenzenesulfonamide ( preparation 274 , 2 . 1 g , 2 . 71 mmol ) at 0 ° c . and the reaction stirred for 15 minutes . semcl was then added ( 1 . 06 ml , 5 . 97 mmol ) and the reaction allowed to warm to room temperature . the reaction was quenched with water , partioned between etoac and brine , the organic layer collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography to afford the title compound ( 760 mg , 43 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 10 ( s , 9h ), − 0 . 05 ( s , 9h ), 0 . 89 ( m , 4h ), 1 . 03 ( t , 3h ), 2 . 88 ( m , 2h ), 3 . 09 ( s , 3h ), 3 . 57 ( t , 2h ), 3 . 73 ( t , 2h ), 4 . 93 ( m , 1h ), 5 . 16 ( m , 1h ), 5 . 31 ( s , 2h ), 5 . 61 ( s , 2h ), 6 . 54 ( m , 1h ), 7 . 10 ( d , 1h ), 7 . 17 ( t , 1h ), 7 . 30 ( t , 1h ), 7 . 42 ( m , 1h ), 7 . 46 ( m , 1h ), 7 . 62 - 7 . 67 ( m , 4h ), 7 . 76 ( m , 1h ), 7 . 95 ( s , 1h ). ms m / z 919 [ m + h ] + the following preparations ( preparations 263 - 268 ) were prepared according to the method described for preparation 131 using either dmf or thf and the appropriate pyrazolopyrimidine as described below . the title compound was prepared according to the methods described for preparations 262 and 258 using n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoro - ethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 273 ). ms m / z 743 [ m + h ] the title compound was prepared according to the methods described for preparations 262 and 258 using n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 275 ). ms m / z 713 [ m + h ] to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethane sulfonamide ( preparation 273 , 300 mg , 0 . 43 mmol ) in dcm ( 10 ml ) at 0 ° c . was added boron tribromide ( 0 . 28 ml , 3 . 02 mmol ). the reaction was stirred at room temperature for 30 minutes before concentrating in vacuo . the residue was partitioned between ethyl acetate and water , the organic layer was collected , dried and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 10 % meoh in dcm to afford the title compound as a white solid ( 250 mg , 87 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 02 ( s , 3h ), 3 . 12 ( s , 3h ), 4 . 11 - 4 . 21 ( m , 2h ), 4 . 80 ( m , 1h ), 4 . 94 ( m , 1h ), 6 . 67 ( m , 1h ), 6 . 76 ( m , 1h ), 6 . 97 ( m , 1h ), 7 . 29 - 7 . 33 ( m , 2h ), 7 . 57 ( m , 1h ), 9 . 58 ( s , 1h ), 10 . 36 ( s , 1h ), 13 . 88 ( s , 1h ). ms m / z 667 [ m + h ] + the following preparations ( preparations 272 - 284 ) were prepared according to the method described for preparation 137 in an organic solvent such as dcm or dmf and using the appropriate pyrazolopyrimidine as described below . the following preparations ( preparations 285 - 293 ) were prepared according to the method described for preparation 140 using either 4m hcl in dioxane or chcl in meoh with the appropriate pyrazolopyrimidine as described below . n -( 2 -((( 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylbenzene sulfonamide ( preparation 301 , 2 . 6 g , 3 . 48 mmol ) was treated with tfa ( 5 ml ) and the reaction stirred at room temperature for 30 minutes before concentrating in vacuo . the residue was diluted with methanol ( 20 ml ), cooled in ice - water and treated with a drop - wise addition of ethylene diamine until the solution became basic . the solution was concentrated in vacuo and purified using silica gel column chromatography eluting with etoac . the residue ( 1 . 7 g , 3 . 19 mmol ) was dissolved in anhydrous thf ( 10 ml ) and 2 , 6 lutidine ( 0 . 55 ml , 4 . 78 mmol ) was added followed by tbdms - triflate ( 0 . 88 ml , 3 . 83 mmol ) at 0 ° c . the reaction was stirred for 18 hours . the reaction was concentrated in vacuo and partitioned between water and ethyl acetate . the organic phase was washed with brine , dried over sodium sulfate , concentrated in vacuo and purified by silica gel column chromatography eluting with 20 % etoac in hexanes to afford the title compound as a white solid ( 1 . 9 g , 92 %). ms m / z 647 [ m + h ] + the following preparations ( preparations 295 - 297 ) were prepared according to the method described for preparation 294 using the appropriate pyrazolopyrimidine as described below . to a solution of 4 -( benzyloxy )- 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethyl - silyl ) ethoxy ) methoxy ) phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine ( preparation 306 , 12 . 5 g , 19 . 75 mmol ) in thf ( 100 ml ) was added 10 % palladium on carbon ( 1 . 5 g ) and the reaction was hydrogenated at 50 psi for 18 hours . the reaction was filtered and the filtrate was concentrated in vacuo . the residue was purified using silica gel column chromatography to afford a white solid . 5 g ( 9 . 22 mmol ) was dissolved in dmf ( 50 ml ) and cooled to 0 ° c . oxalyl chloride ( 7 . 96 ml , 92 mmol ) was added and the reaction stirred at room temperature for 6 hours . the reaction was quenched with water and extracted into etoac . the organic layer was collected , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography to afford the title compound ( 1 . 2 g , 23 %). ms m / z 561 [ m + h ] + a solution of palladium acetate ( 42 mg , 0 . 19 mmol ) and s - phos ( 77 mg , 0 . 19 mmol ) in ethanol ( 10 ml ) was heated at 50 ° c . for 45 minutes after purging with nitrogen ( solution a ). meanwhile a solution of n -( 2 -((( 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 312 , 1 . 7 g , 3 . 77 mmol ) in ethanol ( 30 ml ) was treated with 2 -( 4 -( benzyloxy )- 2 - ethyl - 5 - fluorophenyl )- 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolane ( preparation 323 , 1 . 83 g , 5 . 13 mmol ) and an aqueous solution of potassium phosphate ( 1 . 6 g , 7 . 54 mmol ) in water ( 12 ml ) this solution was purged with nitrogen for 10 minutes ( solution b ). solution a was added to solution b and the mixture heated at 80 ° c . for 18 hours . the reaction was cooled , concentrated in vacuo . the resulting black solid was suspended in ethyl acetate filtered through celite . the filtrate was concentrated in vacuo , the residue was taken up in etoac , washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 40 % etoac in hexanes to afford the title compound as a fluffy white solid ( 1 . 58 g , 65 %). 1 h nmr ( 400 mhz , dmso - ds ): δ ppm 1 . 02 ( m , 3h ), 1 . 55 ( m , 2h ), 1 . 74 ( m , 1h ), 1 . 85 ( m , 1h ), 2 . 00 ( m , 1h ), 2 . 43 ( m , 1h ), 2 . 84 ( m , 2h ), 3 . 07 ( s , 3h ), 3 . 16 ( s , 3h ), 3 . 63 ( m , 1h ), 3 . 93 ( m , 1h ), 4 . 75 ( br m , 1h ), 5 . 00 ( br m , 1h ), 5 . 22 ( s , 2h ), 5 . 85 ( m , 1h ), 7 . 10 ( m , 1h ), 7 . 30 - 7 . 54 ( m , 10h ), 8 . 22 ( s , 1h ), ms m / z 645 [ m + h ] + the following preparations ( preparations 300 - 311 ) were prepared according to the method described for preparation 299 using the appropriate chloro - pyrazolopyrimidine and arylboronic ester as described below . to a stirred solution of 4 , 6 - dichloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine ( wo20131014567a1 , 3 g , 10 . 99 mmol ) in anhydrous n - butanol ( 12 ml ), containing dipea ( 6 . 69 ml , 38 . 45 mmol ) was added n -[ 2 -( aminomethyl ) phenyl ]- n - methylmethane sulphonamide hydrochloride ( wo20101058846a1 , 2 . 76 g , 10 . 98 mmol ) and the reaction was heated in a sealed tube at 90 ° c . for 16 hours . the reaction was concentrated in vacuo and the residue partitioned between water & amp ; ethyl acetate . the organic extracts were washed with brine , dried over sodium sulfate , concentrated in vacuo and triturated with pentane - ether to afford the title compound as an off white solid ( 3 . 5 g , 71 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 17 ( m , 2h ), 1 . 70 ( m , 1h ), 1 . 81 ( m , 1h ), 1 . 98 ( m , 1h ), 2 . 36 ( m , 1h ), 3 . 09 ( s , 3h ), 3 . 25 ( s , 3h ), 3 . 66 ( m , 1h ), 3 . 94 ( m , 1h ), 4 . 60 ( br m , 1h ), 4 . 90 ( br m , 1h ), 5 . 72 ( m , 1h ), 7 . 33 - 7 . 42 ( m , 3h ), 7 . 56 ( m , 1h ), 8 . 22 ( s , 1h ), 9 . 26 ( m , 1h ). the following preparations ( preparations 313 - 321 ) were prepared according to the method described for preparation 312 using 4 , 6 - dichloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine ( wo20131014567a1 ) or other suitable pyrazolopyrimidine and benzylamine as described below . to a suspension of nah ( 0 . 48 g , 20 . 13 mmol ) in thf ( 50 ml ) at 0 ° c . was added benzyl alcohol ( 1 . 98 g , 18 . 30 mol ) slowly . the mixture was allowed to stir for 45 minutes at 0 ° c . followed by the addition of 4 , 6 - dichloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine ( wo20131014567a1 , 5 g , 18 . 30 mmol ). the reaction was stirred at room temperature for 2 hours before being quenched with brine . the solution was extracted into etoac , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography to afford the title compound ( 3 . 4 g , 54 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 56 ( m , 2h ), 1 . 74 - 1 . 78 ( m , 1h ), 1 . 87 - 1 . 91 ( m , 1h ), 1 . 98 - 2 . 02 ( m , 1h ), 2 . 33 - 2 . 43 ( m , 1h ), 3 . 71 ( m , 1h ), 3 . 92 ( m , 1h ), 5 . 62 ( s , 2h ), 5 . 86 ( m , 1h ), 7 . 38 ( m , 3h ), 7 . 55 ( m , 2h ), 8 . 37 ( s , 1h ). ms m / z 345 [ m + h ] + the title compound was prepared according to the method described for preparation 150 using 1 -( benzyloxy )- 4 - bromo - 5 - ethyl - 2 - fluorobenzene ( preparation 324 ). taken on directly to the next step . to a solution of 4 - bromo - 5 - ethyl - 2 - fluorophenol ( wo20131014567a1 , 3 g , 13 . 69 mmol ) in acetone ( 30 ml ) was added benzyl bromide ( 2 . 57 g , 15 . 06 mmol ) and the reaction heated to reflux with potassium carbonate ( 2 . 83 g , 20 . 54 mmol ) for 18 hours . the reaction was filtered , concentrated in vacuo and purified using silica gel column chromatography eluting with 5 % etoac in hexanes to afford the title compound as a colorless oil ( 3 . 20 g , 76 %). 1 h nmr ( 400 mhz , dmso - ds ): δ ppm 1 . 13 ( t , 3h ), 2 . 62 ( q , 2h ), 5 . 18 ( s , 2h ), 7 . 26 - 7 . 51 ( m , 7h ). the title compound was prepared according to the methods described for preparations 323 and 324 using 4 - bromo - 2 - fluoro - 5 -( 2 , 2 , 2 - trifluoroethyl ) phenol ( preparation 326 ). taken on directly to the next step . to a solution of 1 - bromo - 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) benzene ( wo20131014567a1 , 88 . 5 g , 308 . 31 mmol ) at 0 ° c . in dcm ( 2000 ml ), was added boron tribromide ( 204 . 56 ml , 2158 . 17 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was quenched by the addition of cold water dropwise at 0 ° c . the organic layer was separated , the aqueous extracts washed twice with dcm , the organic extracts combined , washed with brine , dried , concentrated in vacuo and triturated with pentane to afford the title compound as a white solid ( 78 g , 93 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 66 - 3 . 74 ( m , 2h ), 7 . 10 ( d , 1h ), 7 . 52 ( d , 1h ), 10 . 50 ( br s , 1h ). a solution of 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl trifluoromethanesulfonate ( preparation 331 , 100 mg , 0 . 15 mmol ), n -( 2 -( aminomethyl ) phenyl )- n -( 2 -( benzyloxy ) ethyl ) methanesulfonamide hydrochloride ( preparation 366 , 64 mg , 0 . 225 mmol ) and triethylamine ( 62 μl , 0 . 45 mmol ) in dmf ( 2 ml ) was heated to between 80 - 90 ° c . for 36 hours . the reaction was cooled and partitioned between etoac ( 50 ml ) and water ( 50 ml ). the organic layer was collected , further washed with water and brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with etoac in heptanes to afford the title compound ( 51 mg , 40 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 00 ( s , 9h ), 0 . 50 ( s , 9h ), 0 . 95 ( m , 3h ), 1 . 05 ( m , 3h ), 1 . 15 ( m , 3h ), 2 . 80 ( m , 2h ), 3 . 10 ( s , 3h ), 3 . 50 ( m , 1h ), 3 . 60 ( m , 4h ), 3 . 80 ( m , 1h ), 3 . 90 ( m , 2h ), 4 . 05 - 4 . 20 ( m , 1h ), 4 . 60 ( m , 2h ), 5 . 15 ( m , 1h ), 5 . 20 ( s , 2h ), 5 . 60 ( s , 2h ), 6 . 15 ( m , 1h ), 6 . 80 ( s , 1h ), 7 . 10 - 7 . 40 ( m , 10h ), 7 . 70 ( m , 1h ), 7 . 90 ( s , 1h ). ms m / z 851 [ m + h ] + triflic anhydride ( 0 . 21 ml , 1 . 25 mmol ) was added dropwise to a solution of 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - ol ( preparation 329 , 495 mg , 0 . 96 mmol ) and pyridine ( 0 . 34 ml , 4 . 2 mmol ) in dcm ( 5 ml ) at 0 ° c . the reaction was stirred at room temperature for 4 hours . the reaction was diluted with water ( 45 ml ), acidified to ph = 3 with citric acid and extracted with etoac ( 2 × 45 ml ). the organic layers were combined , washed with a dilute solution of citric acid at ph = 3 ( 45 ml ), saturated aqueous nahco 3 solution , brine , dried over sodium sulfate and concentrated in vacuo to afford the title compound that was used directly in the next reaction . to a solution of 6 -( 2 - ethyl - 4 -(( 2 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 333 , 8 g , 15 . 5 mmol ) in thf ( 160 ml ) was added tea ( 3 . 13 g , 31 mmol ) dropwise , followed by the addition of acetic anhydride ( 23 . 7 g , 232 . 5 mmol ) dropwise at room temperature . the reaction was heated to 65 ° c . for 18 hours . the reaction was cooled and quenched by the addition of saturated aqueous nahco 3 solution ( 60 ml ), and stirred for 10 hours . the reaction was diluted with water and extracted into etoac . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography to afford the title compound as an oil ( 5 g , 63 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm − 0 . 06 ( s , 9h ), 0 . 0 ( s , 9h ), 0 . 84 - 0 . 88 ( m , 2h ), 0 . 94 - 0 . 98 ( m , 2h ), 1 . 12 - 1 . 16 ( m , 3h ), 2 . 62 - 2 . 64 ( m , 2h ), 3 . 55 - 3 . 59 ( m , 2h ), 3 . 75 - 3 . 79 ( m , 2h ), 5 . 26 ( s , 2h ), 5 . 57 ( s , 2h ), 6 . 42 ( s , 1h ), 6 . 92 - 6 . 96 ( m , 1h ), 6 . 99 ( s , 1h ), 7 . 21 - 7 . 25 ( m , 1h ), 8 . 14 ( s , 1h ), 9 . 15 ( s , 1h ). ms m / z 516 [ m + h ] + the title compound was prepared according to the methods described for preparations 328 and 329 using 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl )- ethoxy ] methoxy } phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 332 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 01 ( s , 9h ), 0 . 91 ( t , 2h ), 1 . 04 ( t , 3h ), 1 . 59 ( m , 2h ), 1 . 71 ( m , 1h ), 2 . 30 ( m , 2h ), 2 . 37 ( m , 2h ), 2 . 70 ( m , 2h ), 3 . 78 ( t , 3h ), 3 . 90 ( d , 1h ), 5 . 36 ( s , 2h ), 6 . 05 ( d , 1h ), 7 . 25 ( d , 1h ), 7 . 35 ( d , 1h ), 8 . 10 ( s , 1h ), 8 . 52 ( s , 1h ). the title compound was prepared according to the methods described for preparations 328 and 329 using 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl )- ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 335 ). used directly in the next reaction . to a stirred solution of 6 -[ 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl )- ethoxy ]- methoxy } phenyl ]- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 147 , 24 g , 50 . 88 mmol ) in anhydrous dcm ( 300 ml ) was added mcpba ( 33 . 52 g , 117 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was quenched with saturated aqueous nahco 3 solution and extracted into dcm . the organic layer was collected , washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 10 % heptanes in etoac to afford the title compound as a yellow solid ( 14 . 5 g , 58 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 0 . 01 ( s , 9h ), 0 . 91 - 0 . 95 ( t , 2h ), 1 . 00 - 1 . 01 ( t , 3h ), 1 . 56 ( s , 2h ), 1 . 66 - 1 . 69 ( m , 1h ), 1 . 95 . 1 . 98 ( m , 2h ), 2 . 28 - 2 . 36 ( m , 3h ), 3 . 69 - 3 . 80 ( m , 3h ), 3 . 71 - 3 . 80 ( m , 3h ), 3 . 86 ( d , 1h ), 5 . 34 ( s , 2h ), 5 . 94 ( d , 1h ), 7 . 16 - 7 . 23 ( m , 2h ), 7 . 94 ( s , 1h ), 8 . 20 ( s , 1h ), 8 . 91 ( s , 1h ). ms m / z 488 [ m + h ] + to a solution of 6 -( 2 - ethyl - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl )- ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 336 , 7 g , 14 mmol ) in dcm ( 100 ml ) was added m - cpba ( 5 . 6 g , 28 mmol ) at room temperature , and the reaction was stirred for 5 hours . the reaction was washed with 10 % aqueous nahso 3 solution and saturated aqueous nahco 3 solution . the organic layer was separated , dried over sodium sulfate and concentrated in vacuo to afford the title compound ( 7 g , 97 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 01 ( s , 9h ), 0 . 09 ( s , 9h ), 0 . 92 - 0 . 96 ( m , 2h ), 1 . 03 - 1 . 07 ( m , 2h ), 1 . 17 - 1 . 21 ( m , 3h ), 2 . 41 - 2 . 56 ( m , 1h ), 2 . 68 - 2 . 81 ( m , 1h ), 3 . 62 - 3 . 66 ( m , 2h ), 3 . 82 - 3 . 87 ( m , 2h ), 5 . 33 ( s , 2h ), 5 . 75 ( s , 2h ), 6 . 99 - 7 . 08 ( m , 1h ), 7 . 12 ( m , 1h ), 7 . 24 ( d , 1h ), 7 . 57 ( d , 1h ), 7 . 57 ( d , 1h ), 8 . 12 ( s , 1h ). ms m / z 516 [ m + h ] + the title compound was prepared according to the method described for preparation 111 using 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy )- methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 337 ). ms m / z 472 [ m + h ] + the title compound was prepared according to the method described for preparation 111 using 6 -( 2 - ethyl - 5 - fluoro - 4 -(( 2 -( trimethylsilyl ) ethoxy ) methoxy ) phenyl )- 1 -(( 2 -( trimethylsilyl )- ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 338 ). ms m / z 534 [ m + h ] + ( preparation 340 , 6 . 7 g , 23 . 67 mmol ) in dmso ( 120 ml ) was added ( 2 -(( 3 - ethyl - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl ) phenoxy ) methoxy ) ethyl ) trimethylsilane ( preparation 343 , 9 . 8 g , 26 . 03 mmol ), potassium phosphate ( 18 . 88 g , 71 . 01 mmol ) and water ( 12 ml ) at room temperature . pd ( pph 3 ) 4 ( 2 . 7 g , 2 . 3 mmol ) was added , the reaction degassed under vacuum and refilled with nitrogen , and heated to 100 ° c . for 18 hours . the reaction was poured into ice water ( 200 ml ) and extracted with etoac . the organic layer was washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography to afford the title compound as a yellow oil ( 6 g , 51 %). taken on directly to the next step . the title compound was prepared according to the method described for preparation 336 using 2 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolane ( wo20131014567a1 ) and 6 - chloro - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 340 ). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 00 ( s , 9h ), 0 . 95 ( m , 2h ), 3 . 62 ( m , 2h ), 3 . 82 ( q , 2h ), 4 . 02 ( s , 3h ), 5 . 80 ( s , 2h ), 7 . 10 ( m , 1h ), 7 . 30 ( m , 1h ), 7 . 60 ( s , 1h ), 8 . 25 ( s , 1h ), 9 . 12 ( s , 1h ). the title compound was prepared according to the method described for preparation 336 using ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 - ethylphenoxy ] methoxy } ethyl )( trimethyl )- silane ( wo20131014567a1 ) and 6 - chloro - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 340 ). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 10 ( s , 9h ), 0 . 10 ( s , 9h ), 0 . 93 - 1 . 00 ( m , 2h ), 1 . 04 - 1 . 08 ( m , 2h ), 1 . 13 - 1 . 22 ( m , 3h ), 2 . 76 ( q , 2h ), 3 . 66 ( m , 2h ), 3 . 91 ( m , 2h ), 5 . 38 ( s , 2h ), 5 . 82 ( s , 2h ), 7 . 21 - 7 . 25 ( m , 2h ), 7 . 56 ( s , 1h ), 8 . 24 ( s , 1h ), 9 . 21 ( s , 1h ). ms m / z 518 [ m + h ] + the title compound was prepared according to the method described for preparation 312 using 4 , 6 - dichloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 341 ) and n -[ 2 -( aminomethyl ) phenyl ]- n - methylmethane - sulfonamide ( wo20101058846a1 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 55 ( m , 2h ), 1 . 69 ( m , 1h ), 1 . 88 - 2 . 00 ( m , 2h ), 2 . 29 ( m , 1h ), 3 . 04 ( s , 1h ), 3 . 27 ( s , 3h ), 3 . 69 ( m , 1h ), 3 . 85 ( m , 1h ), 4 . 55 ( br m , 1h ), 4 . 91 ( br m , 1h ), 5 . 67 ( m , 1h ), 6 . 91 ( s , 1h ), 7 . 32 - 7 . 52 ( m , 4h ), 8 . 24 ( s , 1h ), 8 . 29 ( t , 1h ). ms m / z 449 [ m + h ] + to a solution of 6 - chloro - 1h - pyrazolo [ 4 , 3 - c ] pyridine ( 8 . 5 g , 55 mmol ) in anhydrous thf ( 200 ml ) was added nah ( 60 % dispersion in oil , 2 . 3 g , 58 mmol ) at 0 ° c . after stirring at room temperature for 20 minutes , semcl ( 9 . 67 g , 58 . 06 mmol ) was added dropwise at 0 ° c . the reaction was stirred at room temperature for 2 hours before quenching with water and extracting into etoac . the organic layer was separated , dried over na 2 so 4 and concentrated in vacuo . the residue was purified by silica gel column chromatography to afford the title compound as a yellow oil ( 14 g , 90 %). taken on directly to the next step . the title compound was prepared according to the method described for preparation 149 using 4 , 6 - dichloro - 1h - pyrazolo [ 4 , 3 - c ] pyridine . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 58 - 1 . 61 ( m , 3h ), 1 . 97 - 2 . 03 ( m , 2h ), 2 . 31 - 2 . 34 ( m , 1h ), 3 . 76 - 3 . 80 ( s , 1h ), 3 . 84 - 3 . 91 ( m , 1h ), 5 . 92 - 5 . 95 ( d , 1h ), 8 . 07 ( s , 1h ), 8 . 46 ( s , 1h ). ms m / z 272 [ m + h ] + to a solution of 3 - ethyl - 4 - iodo - 5 - methylphenol ( j . med . chem . ( 2005 ), 48 ( 2 ), 586 - 592 , 500 mg , 1 . 90 mmol ) in acetone ( 20 ml ) was added benzylbromide ( 1 . 43 ml , 2 . 86 mmol ) and potassium carbonate ( 658 mg , 4 . 77 mol ). the reaction was heated to 70 ° c . for 18 hours . the reaction was cooled , filtered and concentrated in vacuo . the residue was dissolved in etoac and washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with hexanes . the residue was dissolved in anhydrous dmso ( 1 . 6 ml ) and bis ( pinacolonato ) diboron ( 1032 mg , 4 . 06 mmol ) and koac ( 543 mg , 5 . 54 mmol ) were added the reaction was purged under argon for 10 minutes before the addition of pd ( dppf ) 2 cl 2 ( 135 mg , 0 . 18 mmol ) followed by degassing for another 10 minutes and then heating to 80 ° c . for 18 hours . the reaction was cooled , concentrated in vacuo and suspended in etoac . the suspension was filtered through celite and the filtrate washed with water , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 5 % etoac in hexanes to afford the title compound . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 1 . 10 ( t , 3h ), 1 . 30 ( s , 12h ), 2 . 28 ( s , 3h ), 2 . 60 ( q , 2h ), 5 . 06 ( s , 2h ), 6 . 63 ( m , 2h ), 7 . 31 - 7 . 44 ( m , 5h ). to a solution of ( 2 -(( 4 - bromo - 3 - ethylphenoxy ) methoxy ) ethyl ) trimethylsilane ( preparation 344 , 300 mg , 0 . 9 mmol ) in dioxane ( 5 ml ) was added bispinacolatodiboron ( 276 mg , 1 . 09 mmol ), pd ( pph 3 ) 4 ( 105 mg , 0 . 09 mmol ) and potassium phosphate ( 384 mg , 1 . 81 mmol ) and the reaction was heated to 80 ° c . for 18 hours . the reaction was cooled and partitioned between water and etoac , eluted though a phase separation cartridge and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 0 - 50 % dcm in heptanes to afford the title compound . taken on directly to the next step . to a solution of 4 - bromo - 3 - ethylphenol ( 9 g , 44 . 8 mmol ) in dcm ( 100 ml ) was added dipea ( 8 . 6 ml , 49 . 3 mmol ) followed by semcl ( 8 . 73 ml , 49 . 3 mmol ) and the reaction was stirred at room temperature for 18 hours . the reaction was washed with water , 1n aqueous hcl solution and saturated aqueous sodium hydrogen carbonate solution , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 3 % etoac in hexanes to afford the title compound . 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 00 ( s , 9h ), 0 . 90 ( m , 2h ), 1 . 25 ( m , 3h ), 2 . 75 ( m , 2h ), 3 . 75 ( m , 2h ), 5 . 20 ( s , 2h ), 6 . 80 ( m , 1h ), 7 . 00 ( d , 1h ), 7 . 40 ( d , 1h ). to a solution of 6 - fluoro - 3 , 4 - dihydro - 2h - isoquinolin - 1 - one ( 13 g , 79 mmol ) in dmso ( 150 ml ) was added 4 - methylimidazole ( 7 . 8 g , 95 mmol ) followed by cesium carbonate ( 38 g , 118 . 5 mmol ) and the reaction was heated to 125 ° c . for 18 hours . the reaction was cooled and extracted into chloroform / isopropanol ( v : v 3 : 1 , 500 ml ) three times . the organic layers were combined , washed with brine , dried over sodium sulfate and concentrated in vacuo . a portion of the residue ( 9 g , 39 . 6 mmol ) was dissolved in thf and cooled to 0 ° c . lialh 4 ( 3 g , 79 . 2 mmol ) was added portionwise , and the reaction heated to 60 ° c . for 18 hours . the reaction was cooled and quenched by the addition of 10 % naoh solution ( 6 ml ), before filtration and concentration in vacuo . the residue was purified by silica gel column chromatography eluting with 30 - 100 % etoac in petroleum ether followed by the addition of 2n hcl in etoac . the resulting precipitate was filtered to afford the title compound as the hydrochloride salt ( 11 . 6 g , 42 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 34 ( s , 3h ), 3 . 08 ( m , 2h ), 3 . 37 ( m , 2h ), 4 . 30 ( m , 2h ), 7 . 48 ( m , 1h ), 7 . 62 - 7 . 68 ( m , 2h ), 8 . 00 ( s , 1h ), 9 . 61 ( s , 1h ), 9 . 91 ( br s , 2h ). to a solution of 1 , 2 , 3 , 4 - tetrahydro - 2 -( 2 , 2 , 2 - trufluoroacetyl )- 7 - isoquinoline sulfonyl chloride ( 400 mg , 1 . 2 mmol ) in meoh ( 5 ml ) was added 2 -( pyrrolidin - 1 - yl ) ethanamine in excess and the reaction stirred at room temperature for 30 minutes . water ( 1 ml ) followed by potassium carbonate ( 150 mg , 1 . 4 mmol ) were added and the reaction stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and the residue dissolved in dcm . the suspension was filtered and the filtrate purified by silica gel column chromatography eluting with 10 - 100 % ( 90 : 10 : 1 dcm : meoh : nh 3 ) in dcm to afford the title compound ( 130 mg , 35 %). ms m / z 310 [ m + h ] + to a solution of 2 -( tert - butoxycarbonyl )- 1 , 2 , 3 , 4 - tetrahydroisoquinoline - 5 - carboxylic acid ( 200 mg , 0 . 721 mmol ) and dipea ( 87 μl , 0 . 793 mmol ) in dcm ( 10 ml ) was added hbtu ( 301 mg , 0 . 793 mmol ) followed by a solution of benzylamine ( 151 μl 0 . 865 mmol ) in dcm ( 5 ml ) and the reaction was stirred at room temperature for 72 hours . the reaction was washed with water ( 1 ml ), 1n hcl ( aq ) ( 1 ml ) and 1n naoh ( aq ) ( 1 ml ). the organic layer was dried over sodium sulfate and concentrated in vacuo . the residue was dissolved in meoh ( 5 ml ) and 4n hcl in dioxane ( 3 ml ) was added . the reaction was stirred at room temperature for 18 hours . the reaction was concentrated in vacuo and triturated with diethylether to afford the title compound as the hydrochloride salt ( 200 mg , quant .). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 09 ( t , 2h ), 3 . 33 ( m , 2h ), 4 . 28 ( t , 2h ), 4 . 44 ( d , 2h ), 5 . 19 ( s , 2h ), 5 . 95 ( d , 1h ), 7 . 23 - 7 . 42 ( m , 8h ), 8 . 93 ( t , 1h ), 9 . 49 ( br s , 2h ). to a solution of 1 - benzhydryl - 3 - azetidinyl methanesulfonate ( 44 . 6 g , 0 . 147 mol ) and 2 - chloro - 5 - cyanophenol ( 22 . 6 g , 0 . 147 mol ) in mecn ( 600 ml ) was added cs 2 co 3 ( 62 . 3 g , 0 . 19 mol ). the reaction was stirred at 80 ° c . for 24 hours . the reaction was filtered , and the filtrate was concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 5 - 20 % etoac in petroleum ether . the residue was dissolved in dichloroethane ( 550 ml ) and potassium carbonate ( 66 . 4 g , 0 . 48 mmol ) followed by ace - cl ( 20 . 5 g , 0 . 14 mmol ) was added . the reaction was heated to reflux for 2 hours . the reaction was concentrated in vacuo and the residue was recrystallized with meoh to afford the title compound as the hydrochloride salt ( 13 . 8 g , 59 %). 1 h nmr ( 400 mhz , meod ): δ ppm 4 . 21 - 4 . 25 ( m , 2h ), 4 . 61 - 4 . 66 ( m , 2h ), 5 . 25 - 5 . 31 ( m , 1h ), 7 . 30 ( s , 1h ), 7 . 40 - 7 . 43 ( d , 1h ), 7 . 63 - 7 . 65 ( d , 1h ). to a solution of tert - butyl 2 -( n - propylmethylsulfonamido ) benzylcarbamate ( preparation 351 , 265 mg , 0 . 77 mmol ) in dcm ( 2 ml ) was added tfa ( 0 . 5 ml ) and the reaction stirred at room temperature for 1 hour . the reaction was diluted with dcm and washed with a 1 : 1 mixture of 880 nh 3 in water ( 20 ml ). the organic layer was collected , dried over magnesium sulfate and concentrated in vacuo to afford the title compound as the trifluoroacetate salt ( 172 mg , 92 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 89 ( t , 3h ), 1 . 41 - 1 . 55 ( m , 2h ), 2 . 96 ( s , 3h ), 3 . 39 - 3 . 46 ( m , 1h ), 3 . 62 - 3 . 69 ( m , 1h ), 3 . 83 - 3 . 87 ( m , 1h ), 4 . 10 - 4 . 20 ( m , 1h ), 7 . 19 - 7 . 22 ( m , 1h ), 7 . 28 - 7 . 32 ( m , 1h ), 7 . 36 - 7 . 40 ( m , 1h ), 7 . 57 ( d , 1h ). the title compound was prepared according to the method described by preparation 349 using tert - butyl 2 -( n - butylmethylsulfonamido ) benzylcarbamate ( preparation 354 ) and isolated as the trifluoroacetate salt . 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 87 ( t , 3h ), 1 . 26 - 1 . 52 ( m , 4h ), 2 . 95 ( s , 3h ), 3 . 41 - 3 . 48 ( m , 1h ), 3 . 67 - 3 . 75 ( m , 1h ), 3 . 83 - 3 . 86 ( m , 1h ), 4 . 10 - 4 . 14 ( m , 1h ), 7 . 19 - 7 . 21 ( m , 1h ), 7 . 28 - 7 . 32 ( m , 1h ), 7 . 37 - 7 . 41 ( m , 1h ), 7 . 58 ( d , 1h ). the title compound was prepared according to the method described by preparation 213 using n -( 2 - cyanophenyl )- n - propylmethanesulfonamide ( preparation 352 ). taken on directly to the next step . to a solution of n -( 2 - cyanophenyl ) methanesulfonamide ( preparation 223 , 500 mg , 2 . 55 mmol ) in nmp ( 10 ml ) was added sodium hydride ( 148 mg , 3 . 83 mmol ) and the reaction stirred for 30 minutes at room temperature . propyl iodide ( 1 . 74 ml , 3 . 83 mmol ) was added and the reaction was stirred at room temperature for 18 hours . the reaction was quenched by the addition of water and extracted into etoac . the organic layer was collected , washed with water , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 30 % etoac in heptanes to afford the title compound ( 505 mg , 83 %). 1 h nmr ( 400 mhz , cdcl 3 ): δ ppm 0 . 94 ( t , 3h ), 1 . 48 - 1 . 60 ( m , 2h ), 3 . 11 ( s , 3h ), 3 . 71 ( t , 2h ), 7 . 46 - 7 . 54 ( m , 2h ), 7 . 65 - 7 . 69 ( m , 1h ), 7 . 72 - 7 . 74 ( m , 1h ). sodium hydride ( 76 mg , 1 . 92 mmol ) was added to a solution of tert - butyl 2 -( n - methylmethylsulfonamido ) benzylcarbamate ( wo20101058846a1 , 200 mg , 0 . 64 mmol ) in nmp and the reaction was stirred at 0 ° c . for 30 minutes . 2 - morpholinoethanamine ( 226 mg , 0 . 96 ml ) was added and the reaction stirred at room temperature for 18 hours . the reaction was quenched by the addition of water and extracted into etoac . the organic layer was collected , dried and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 100 : 10 : 1 dcm : meoh : tea . the residue was dissolved in dcm ( 2 ml ) and tfa ( 1 ml ) was added . the reaction was stirred at room temperature for 1 hour . the reaction was concentrated in vacuo to afford the title compound as the trifluoroacetate salt . ms m / z 328 [ m + h ] + the following preparations ( preparations 354 - 359 ) were prepared according to the method described by preparation 353 using the appropriate sulphonamide and alkyl halide as described below . the compounds were isolated according to the described experimental or by dissolving in dcm ( 20 ml ) and washing with a 1 : 1 mixture of ammonium hydroxide : water . the organic layer was collected , dried over magnesium sulfate and concentrated in vacuo to afford the title compound that was used in the next reaction directly . the following preparations ( preparations 360 - 364 ) were prepared according to the methods described by preparations 351 and 352 using the appropriate sulphonamide and alkyl halide as described below : the title compound was prepared according to the method described for preparation 223 using methyl iodide and n -( 4 - methyl - 2 - cyanophenyl )- n -( methylsulfonyl ) methanesulfonamide ( preparation 242 ). taken on directly to the next step . the title compound was prepared according to the methods described for preparations 222 , 213 and 211 using 2 -( benzyloxy ) ethanol and n -( 2 - cyanophenyl ) methanesulfonamide . ms m / z 335 [ m + h ] + to a solution of 2 -( 3 , 4 - dimethoxyphenyl )- n -( 2 -( n - methylmethylsulfonamido ) benzyl ) acetamide ( preparation 368 , 800 mg , 2 . 03 mmol ) in thf ( 15 ml ) was added borane - dimethylsulfide ( 2m in thf , 2 . 55 ml , 5 . 10 mmol ) and the reaction was heated to reflux for 2 . 5 hours . the reaction was cooled , concentrated in vacuo and the residue dissolved in methanol ( 12 ml ). the solution was treated with 6n hcl ( 8 ml ) and heated to reflux for 2 hours . the reaction was concentrated in vacuo and the residue basified with 3n naoh solution . the aqueous layer was extracted into 10 % meoh / dcm , the organic extracts were washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified using silica gel column chromatography eluting with 7 % meoh in dcm to afford the title compound ( 390 mg , 51 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 63 - 2 . 71 ( m , 4h ), 3 . 04 ( s , 3h ), 3 . 17 ( s , 3h ), 3 . 71 ( s , 6h ), 3 . 80 ( br s , 2h ), 6 . 68 - 6 . 83 ( m , 3h ), 7 . 28 - 7 . 51 ( m , 4h ). ms m / z 379 [ m + h ] + to a solution of n -[ 2 -( aminomethyl ) phenyl ]- n - methylmethanesulfonamide ( wo 2010 / 058846a1 , 1 g , 3 . 64 mmol ) and 2 -( 3 , 4 - dimethoxyphenyl ) acetic acid ( 786 mg , 4 . 00 mmol ) in thf ( 20 ml ) was added propylphosphonic anhydride ( 2 . 9 g , 9 . 11 mmol ) followed by dipea ( 2 . 21 ml , 12 . 68 mmol ) and the reaction was stirred at room temperature for 14 hours . the reaction was concentrated in vacuo and the residue was partitioned between etoac and saturated aqueous sodium bicarbonate solution . the organic layer was collected , washed with brine , dried , concentrated in vacuo and purified using silica gel column chromatography eluting with 4 % meoh in dcm to afford the title compound as a white solid ( 540 mg , 38 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 3 . 05 ( s , 3h ), 3 . 14 ( s , 3h ), 3 . 40 ( s , 2h ), 3 . 71 ( s , 6h ), 4 . 26 ( br s , 1h ), 4 . 46 ( br s , 1h ), 6 . 77 - 6 . 88 ( m , 3h ), 7 . 26 - 7 . 34 ( m , 3h ), 7 . 45 - 7 . 47 ( m , 1h ), 8 . 34 ( t , 1h ). ms m / z 393 [ m + h ] + the title compound was prepared according to the methods described for preparations 368 and 367 using 2 -( 4 -( methylsulfonamido ) phenyl ) acetic acid . 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm 2 . 66 - 2 . 73 ( m , 4h ), 2 . 92 ( s , 3h ), 3 . 04 ( s , 3h ), 3 . 13 ( s , 3h ), 3 . 80 ( br s , 2h ), 7 . 09 - 7 . 17 ( m , 4h ), 7 . 29 - 7 . 34 ( m , 2h ), 7 . 42 - 7 . 49 ( m , 2h ), 9 . 52 ( br s , 1h ). ms m / z 412 [ m + h ] + the title compound was prepared according to the method described for example 157 using n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 273 ) and tert - butyl 2 - iodo - 6 , 7 - dihydro - 1h - imidazo [ 4 , 5 - c ] pyridine - 5 ( 4h )- carboxylate ( wo20131014567a1 ) using hcl in dioxane for the deprotection step . ms m / z 690 [ m + h ] + to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 -( hydrazinecarbonyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl )- 4 - methoxyphenyl )- n - methylmethanesulfonamide ( preparation 375 , 450 mg , 0 . 59 mol ) in butanol ( 2 ml ) was added tert - butyl 4 - cyanopiperidine - 1 - carboxylate ( 624 mg , 2 . 97 mmol ) and the reaction was heated to 150 ° c . under microwave irradiation for 50 minutes . the reaction was cooled , filtered and concentrated in vacuo . the residue was purified using preparative hplc . the residue was treated with tfa ( 2 ml ) and stirred at room temperature for 30 minutes . the reaction was concentrated in vacuo , dissolved in meoh ( 5 ml ) and cooled in ice water . ethylene diamine was added dropwise until the solution was basic , with stirring for 1 hour . the solution was concentrated in vacuo and extracted into 20 % ipa in dcm . the organic layer was washed with water , dried over sodium sulfate and concentrated in vacuo to afford the title compound that was used directly in the next reaction . ms m / z 719 [ m + h ] + the title compound was prepared according to the method described by preparation 371 using n -( 2 -((( 3 - cyano - 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoro - ethyl ) phenyl )- 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino ) methyl ) phenyl )- n - methylmethanesulfonamide ( preparation 377 ) and tert - butyl 4 -( hydrazinecarbonyl ) piperidine - 1 - carboxylate in the presence of potassium carbonate . ms m / z 689 [ m + h ] + the title compound may be prepared according to the method described for preparation 11 using n -[ 2 -({[ 6 -( 2 - ethyl - 5 - fluoro - 4 -{[ 2 -( trimethylsilyl ) ethoxy ] meth - oxy } phenyl )- 3 - iodo - 1 -{[ 2 -( trimethylsilyl )- ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ]- amino } methyl ) phenyl ]- n - methylmethanesulfonamide ( preparation 79 ). taken on directly to the next step . to a solution of 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 4 -({ 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } amino )- 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine - 3 - carboxylic acid ( preparation 12 , 0 . 55 g , 0 . 66 mmol ) in meoh / toluene ( 15 ml ) was added 2m trimethylsilyldiazomethane in thf ( 0 . 997 ml , 1 . 99 mmol ) dropwise at 0 ° c . the reaction was stirred for 2 hours at room temperature . the reaction was concentrated in vacuo and the residue was purified using silica gel column chromatography eluting with 8 % meoh in dcm . the residue was dissolved in meoh ( 5 ml ) and hydrazine monohydrate ( 40 . 12 mg , 0 . 80 mmol ) was added . the reaction was heated to reflux for 18 hours . the reaction was concentrated in vacuo and the residue was purified using neutral alumina column chromatography eluting with 50 % etoac in hexanes to afford the title compound ( 297 mg , 66 %). ms m / z 842 [ m + h ] + the title compound was prepared according to the method described by preparation 374 using 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 4 -(( 5 - methoxy - 2 -( n - methylmethylsulfonamido ) benzyl ) amino )- 1 -(( 2 -( trimethylsilyl ) ethoxy )- methyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidine - 3 - carboxylic acid ( preparation 269 ). ms m / z 757 [ m + h ] + the - was prepared according to the methods described for preparations 11 and 374 using n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - tri - fluoroethyl ) phenyl )- 3 - iodo - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ) methyl ) phenyl )- n - methyl - methanesulfonamide ( preparation 378 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 11 ( s , 9h ), 0 . 83 ( m , 2h ), 1 . 23 ( br s , 2h ), 3 . 05 ( s , 3h ), 3 . 11 ( s , 3h ), 3 . 57 ( m , 2h ), 3 . 77 ( m , 2h ), 3 . 86 ( s , 3h ), 4 . 68 ( m , 2h ), 4 . 80 ( br m , 1h ), 4 . 90 ( br m , 1h ), 5 . 72 ( s , 2h ), 7 . 08 ( s , 1h ), 7 . 20 - 7 . 51 ( m , 6h ), 9 . 68 ( t , 1h ), 10 . 17 ( m , 1h ). ms m / z 726 [ m + h ] + to a solution of n -( 2 -((( 6 -( 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - pyrazolo [ 4 , 3 - c ] pyridin - 4 - yl ) amino ) methyl )- phenyl )- n - methylmethanesulfonamide ( preparation 378 , 1 . 2 g , 1 . 51 mmol ) in dmf ( 10 ml ), was added zinc cyanide ( 0 . 19 g , 1 . 66 mmol ) and pd ( pph 3 ) 4 ( 0 . 05 mg , 0 . 04 mmol ). the reaction was degassed with nitrogen and heated to 120 ° c . under microwave irradiation for 20 minutes . the reaction was quenched with water and extracted into ethyl acetate . the organic extracts were dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 48 % etoac in hexanes to afford the title compound ( 610 mg , 58 %). 1 h nmr ( 400 mhz , dmso - d 6 ): 5 ppm − 0 . 07 ( s , 9h ), 0 . 86 ( t , 2h ), 3 . 08 ( s , 3h ), 3 . 17 ( s , 3h ), 3 . 65 ( t , 2h ), 3 . 88 ( s , 3h ), 4 . 22 ( m , 2h ), 4 . 89 ( br m , 1h ), 5 . 00 ( br m , 1h ), 5 . 75 ( s , 2h ), 7 . 24 - 7 . 41 ( m , 4h ), 7 . 55 - 7 . 59 ( m , 2h ), 8 . 29 ( t , 1h ). ms m / z 694 [ m + h ] + the title compound was prepared according to the method described for preparation 61 using 6 -[ 5 - fluoro - 4 - methoxy - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl ]- 3 - iodo - 1 -{([ 2 -( trimethylsilyl ) ethoxy ]- methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine ( preparation 111 ) and 4 - nitrophenyl { 2 -[ methyl ( methylsulfonyl ) amino ] benzyl } carbamate ( preparation 166 ). ms m / z 794 [ m + h ] + the title compound was prepared according to the method described for preparation 61 using 6 -[ 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -{[ 2 -( trimethylsilyl ) ethoxy ] methoxy } phenyl ]- 3 - iodo - 1 -{[ 2 -( trimethylsilyl ) ethoxy ] methyl }- 1h - pyrazolo [ 4 , 3 - c ] pyridine 5 - oxide ( preparation 114 ) and 4 - nitrophenyl ( 2 -( methylthio ) ethyl ) carbamate ( preparation 385 ). ms m / z 787 [ m + h ] + the title compound was prepared according to the method described for preparation 131 using racemic n -( 3 -(( tert - butyldimethylsilyl ) oxy )- 2 - methylpropyl )- 6 -( 4 -(( tert - butyldimethylsilyl )- oxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 3 - iodo - 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - amine ( preparation 381 ). ms m / z 786 [ m + h ] + the title compound was prepared according to the method described for preparation 137 using racemic n -( 3 -(( tert - butyldimethylsilyl ) oxy )- 2 - methylpropyl )- 6 -( 4 -(( tert - butyldimethylsilyl ) oxy )- 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl ) phenyl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - amine ( preparation 382 ). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 01 ( s , 6h ), 0 . 22 ( s , 6h ), 0 . 79 ( m , 9h ), 0 . 98 ( m , 12h ), 2 . 04 ( m , 1h ), 3 . 48 ( m , 1h ), 3 . 56 ( m , 2h ), 3 . 68 ( m , 1h ), 4 . 45 ( m , 2h ), 6 . 71 ( m , 1h ), 7 . 13 ( m , 1h ), 7 . 81 ( m , 1h ), 13 . 88 ( s , 1h ). ms m / z 754 [ m + h ] + the title compound was prepared according to the methods described by preparations 142 and 141 using racemic 3 -(( 6 -( 5 - fluoro - 2 -( 2 , 2 , 2 - trifluoroethyl )- 4 -(( 2 -( trimethylsilyl )- ethoxy ) methoxy ) phenyl )- 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino )- 2 - methylpropan - 1 - ol ( preparation 383 ). ms m / z 626 [ m − h ]− the title compound was prepared according to the method described for preparation 299 using racemic 3 -(( 6 - chloro - 1 -( tetrahydro - 2h - pyran - 2 - yl )- 1h - pyrazolo [ 3 , 4 - d ] pyrimidin - 4 - yl ) amino )- 2 - methylprop - an - 1 - ol ( preparation 384 ) and ( 2 -{[ 2 - fluoro - 4 -( 4 , 4 , 5 , 5 - tetramethyl - 1 , 3 , 2 - dioxaborolan - 2 - yl )- 5 -( 2 , 2 , 2 - trifluoroethyl ) phenoxy ]- methoxy } ethyl ) ( trimethyl ) silane ( preparation 150 ). ms m / z 614 [ m + h ] + the title compound was prepared according to the method described by preparation 299 using racemic 3 - amino - 2 - methylpropan - 1 - ol . ms m / z 326 [ m + h ] + the title compound was prepared according to the method described for preparation 156 using 2 -( methylthio ) ethanamine . taken on directly to the next step . to solution of 4 , 5 - dimethyl - 1 -(( 2 -( trimethylsilyl ) ethoxy ) methyl )- 1h - imidazole ( preparation 387 , 270 mg , 1 . 19 mmol ) in anhydrous thf ( 3 ml ) at − 78 ° c ., was added butyllithium ( 0 . 54 ml , 1 . 31 mmol ) dropwise . the reaction was kept at − 78 ° c . for 15 minutes before the addition of carbon tetrabromide ( 474 mg , 1 . 43 mmol ) in thf ( 2 ml ). the reaction was warmed to room temperature before quenching with ammonium chloride and extracting into etoac . the organic layer was collected , washed with water , brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 3 % meoh in dcm to afford the title compound as a colorless oil ( 220 mg , 60 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 01 ( s , 9h ), 0 . 87 ( t , 2h ), 2 . 01 ( s , 3h ), 2 . 14 ( s , 3h ), 3 . 51 ( t , 2h ), 5 . 18 ( s , 2h ). a suspension of nah ( 124 mg , 3 . 12 mmol ) in dmf ( 3 ml ) was added a solution of 4 , 5 - dimethyl - 1h - imidazole ( 200 mg , 2 . 08 mmol ) in dmf ( 2 ml ) at 0 ° c . the suspension was stirred for 15 minutes before the dropwise addition of sem chloride ( 0 . 44 ml , 2 . 49 mmol ). the reaction was stirred at room temperature for 1 hour , then partitioned between ethyl acetate and water . the combined organic extracts were washed with brine , dried over sodium sulfate and concentrated in vacuo . the residue was purified by silica gel column chromatography eluting with 5 % meoh in dcm to afford the title compound as a colorless oil ( 270 mg , 57 %). 1 h nmr ( 400 mhz , dmso - d 6 ): δ ppm − 0 . 01 ( s , 9h ), 0 . 82 ( t , 2h ), 2 . 01 ( s , 3h ), 2 . 09 ( s , 3h ), 3 . 43 ( t , 2h ), 5 . 18 ( s , 2h ), 7 . 52 ( s , 1h ). test article was solubilized in dimethyl sulfoxide ( dmso ) to a stock concentration of 30 mm . an 11 - point half log dilution series was created in dmso with a top concentration of 600 μm . the test compound plate also contained positive control wells containing a known inhibitor to define 100 % inhibition and negative control wells containing dmso to define no inhibition . the compound plates were diluted 1 to 60 resulting in a top final assay compound concentration of 10 μm and a 2 % dmso concentration . test article and assay controls were added to a 384 - well plate . reaction mixtures contained 20 mm hepes , ph 7 . 4 , 10 mm magnesium chloride , 0 . 01 % bovine serum albumin ( bsa ), 0 . 0005 % tween 20 , 1 mm atp and 1 μm peptide substrate . the jak1 and tyk2 assays contained 1 μm of the irstide peptide ( 5fam - kksrgdymtmqid ) and the jak2 and jak3 assays contained 1 μm of the jaktide peptide ( fitc - kggeeeeyfelvkk ). the assays were initiated by the addition of 20 nm jak1 , 1 nm jak2 , 1 nm jak3 or 1 nm tyk2 enzyme and were incubated at room temperature for three hours for jak1 , 60 minutes for jak2 , 75 minutes for jak3 or 135 minutes for tyk2 . enzyme concentrations and incubation times were optimized for each new enzyme preps and were modified slightly over time to ensure 20 %- 30 % phosphorylation . the assays were stopped with a final concentration of 10 mm edta , 0 . 1 % coating reagent and 100 mm hepes , ph = 7 . 4 . the assay plates were placed on a caliper life science lab chip 3000 ( lc3000 ) instrument , and each well was sampled using appropriate separation conditions to measure the unphosphorylated and phosphorylated peptide . an assay measuring the efficacy of jak inhibitors on the functional response of recombinant human interferon γ ( rhifnγ ) stimulated stat - 3 phosphorylation in the a549 human epithelial cell line . a549 cells ( atcc # ccl - 185 ), were plated at 30 , 000 cells / well in 96 well flat bottomed tissue culture plates ( bd # 353072 ) in 200 μl of growth medium ( dmem , pfizer media prep , with 10 % fetal bovine serum , sigma # f4135 , 2 mm l - glutamine , pfizer media prep , 100 u / ml penicillin , pfizer media prep , and 200 μg / ml streptomycin , pfizer media prep ), and cultured at 37 ° c ., 5 % co 2 incubator for 18 hours . growth medium was removed by vacuum aspiration ( v & amp ; p scientific # vp187 bp - 60 ), and 90 μl of pre - warmed assay medium ( dmem with 0 . 2 % bsa , miltenyi # 130 - 091 - 376 ) was added to each well and incubated for 15 minutes at 37 ° c . 10 μl of vehicle control or test compound ( final concentration range of 0 . 3 nm to 10 μm with 0 . 1 % dmso ) was added to the cells . plates were incubated at 37 ° c . for 1 hour . after compound incubation , 10 μl of 220 ng / ml recombinant human ifnγ ( r & amp ; d systems # 285 - if , final rhifnγ concentration of 20 ng / ml ) was added to the cells and plates were incubated for 30 minutes at 37 ° c . wells containing a549 cells , medium with 0 . 1 % dmso and no rhifnγ were used as background controls . after rhifnγ stimulation , media was aspirated from each well and 35 μl / well of iced - cold msd lysis buffer containing protease and phosphatase inhibitors from phospho - stat3 tyr705 assay kit ( meso - scale discovery # k150did ) was added to each well . plates were incubated at 4 ° c . with shaking for 30 minutes . cell lysates were assayed following the msd phospho - stat3 tyr705 assay kit protocol to detect pstat3 . data were collected and transformed into percent inhibition and calculated using the following formula : data were graphically displayed as percent inhibition using graphpad prism 4 . 0 , and ic 50 curves were fitted using a point to point analysis . an assay measuring the efficacy of jak inhibitors on the functional response of recombinant human interleukin - 2 ( rhil - 2 ) stimulated stat5 phosphorylation in isolated human t cells . human whole blood from individual donors was collected from the phlebotomy unit on - site . peripheral venous blood ( 30 - 60 ml ) from healthy volunteers of either sex was used as the source of t cells . t cell isolation from venous whole blood is routinely performed in a class ii microbiological safety cabinet . each sample was collected into between 3 and 6 10 ml sodium heparin vacutainer tubes ( bd # 367874 ). the blood was poured into sterile 50 ml conicals ( corning # 430828 ) and incubated with the t cell rosette sep cocktail ( stemcell technologies # 15061 ) at 50 μl / ml antibody / blood ratio for 20 min . with shaking at room temperature . the blood / antibody mixture was then diluted 1 : 2 with pbs ( pfizer media prep )/ 2 % fbs ( sigma # f4135 ) and 30 ml of the mixture was layered onto 15 ml ficoll - hypaque ( ge healthcare # 17 - 1440 - 03 ) in 50 ml conical tubes . the tubes were then centrifuged at 1200 × g for 20 min . at room temperature with no brake . following centrifugation the t cells formed a buffy coat between the ficoll - hypaque and plasma layers . the plasma above the buffy coats was removed to within 5 mm of the buffy coat using a sterile pasteur pipette . the buffy coats were then collected into fresh sterile 50 ml conical tubes containing 25 ml pbs / 2 % fbs ( 2 buffy coats per 50 ml conical ). pbs / 2 % fbs was added to the buffy coat cells such that the final volume in the tube was 50 ml . the tubes were then centrifuged at 200 × g for 15 min . at room temperature . the supematant was discarded and the pellet re - suspended in 10 - 20 ml of dmem ( pfizer media prep ) assay media / 0 . 2 % bsa ( miltenyi # 130 - 091 - 376 ). a differential cell count was performed using a haemacytometer and cells were diluted to 1 . 1 × 10 6 t cells / ml in dmem / 0 . 2 % bsa media . compounds ( 10 mm - 0 . 3 μm ) were diluted with hanks balanced salt solution ( hbss ) ( sigma # h6648 ) at 1 : 100 dilution . immediately following cell isolation and compound dilution , 90 μl / well of t cells (˜ 1 × 10 6 / ml ) in assay medium ( dmem + 0 . 2 % bsa ) was added to the vwr deep well v bottom plate (# 3906 - 520 - 300 ). 10 μl / well of compound ( final concentration range of 10 μm - 0 . 3 nm with 0 . 1 % dmso ) or 0 . 1 % dmso in hbss as controls was added to the appropriate wells . plates were incubated for 1 hour at 37 ° c ., 5 % co 2 incubator . 10 μl of 3 . 3 μg / ml rhil - 2 ( r & amp ; d systems # 202 - il ) was added to the cells ( 300 ng / ml final assay concentration ). wells containing t cells , medium with 0 . 1 % dmso and no rhil - 2 were used as background controls . plates were incubated for 15 min at 37 ° c . after rhil - 2 stimulation , 800 μl of cold pbs / 0 . 1 % bsa was added and the plates were centrifuged at 1400 rpm for 5 min at 4 ° c . supematant was aspirated and 100 μl of ice - cold msd lysis buffer containing protease and phosphatase inhibitors from phospho - stat5a / b tyr694 assay kit ( meso - scale discovery # k150igd ) was added to the cell pellet . plates were shaken for 30 min . at 4 ° c . and then frozen overnight . the following day , cell lysates were assayed using the msd elisa kit protocol for detection of pstat5 . data were collected and transformed into percent inhibition and calculated using the following formula : data were graphically displayed as percent inhibition using graphpad prism 4 . 0 , and ic 50 curves were fitted using a point to point analysis .

a compound having the structure : or a pharmaceutically acceptable salt thereof , or a pharmaceutically acceptable solvate of said compound or pharmaceutically acceptable salt , wherein a and a ′ are independently c or n , where c may be unsubstituted or substituted by halo or c 1 - c 6 alkyl ; r and r 0 are independently selected from the group consisting of h , c 1 - c 6 alkyl , hydroxy , phenyl , and — n — w , where w is c 3 - c 8 cycloalkyl , phenyl , naphthyl , 5 - or 6 - membered heteroaryl or heterocyclic containing 1 - 3 n , s and / or o atoms , — so 2 — r ′, — nhso 2 — r ′, — nr ″ so 2 — r ′ and sr ′, where r ′ and r ″ are independently c 1 - c 6 alkyl or c 3 - c 8 cycloalkyl , etc . ; wherein each of said alkyl , cycloalkyl , heterocyclic , phenyl , naphthyl or heteroaryl may be unsubstituted or substituted by phenyl , heteroaryl , etc . ; or , r and r 0 and the n atom to which they are bonded together form a monocyclic or bicyclic heterocyclic ring which may be unsubstituted or substituted by halo , hydroxy , heteroaryl , c 1 - c 6 alkyl , c 1 - c 6 alkoxy , etc ., or — n — w , where w is c 3 - c 8 cycloalkyl , phenyl , etc . ; r 1 is h , halo or cyano ; r 2 and r 2 ′ are independently h , c 1 - c 6 alkyl , cyano , c 1 - c 6 alkoxy , c 1 - c 6 alkylthio , or c 3 - c 8 cycloalkyl where alkyl , alkoxy , or cycloalkyl is optionally substituted by one or more fluorine atoms ; x is a bond , — co —, — conh —, — so 2 —, — sonh —, or — m —; r 3 is h , c 1 - c 4 alkyl , phenyl , naphthyl , 5 - or 6 - membered heteroaryl or heterocyclic containing 1 - 3 n atoms , a 5 - membered heteroaryl or heterocyclic , etc ., or 2 o or s atoms and 0 - 2 n atoms ; wherein each of said phenyl , naphthyl , heteroaryl or heterocyclic is optionally substituted by alkyl , 1 substituent — y — r 4 and / or 1 - 4 substituents each independently selected from r 5 ; with the proviso that when x is — co — or — so 2 —, r 3 is not h ; y is a bond , — m — or — o —; r 4 is h , c 1 - c 6 alkyl , c 3 - c 8 cycloalkyl , halo , oxo , — or 6 , — nr 7 r 8 , — sr 6 , — sor 9 , — so 2 r 9 , — cor 6 , — ocor 6 , — coor 6 , — nr 6 cor 6 , — conr 7 r 8 , etc . ; phenyl or naphthyl , said phenyl and naphthyl being optionally substituted with 1 - 5 substituents selected from c 1 - c 6 alkyl , c 3 - c 8 cycloalkyl , halo , cyano , — or 6 , — nr 7 r 8 , etc . ; or a 3 to 8 - membered saturated or partially unsaturated monocyclic heteroaryl , etc . ; r 6 is h , c 1 - c 6 alkyl or c 3 - c 8 cycloalkyl , etc . ; r 7 and r 8 are each independently h , c 1 - c 6 alkyl or c 3 - c 8 cycloalkyl or are taken together with the nitrogen atom to which they are attached to form a 4 -, 5 - or 6 - membered saturated heterocyclic ring containing 1 - 2 nitrogen atoms or 1 nitrogen and 1 oxygen atom , said c 1 - c 6 alkyl is optionally substituted by c 3 - c 8 cycloalkyl , halo , etc ., and said heterocyclic ring being optionally substituted by one or more c 1 - c 6 alkyl or c 3 - c 8 cycloalkyl groups ; r 9 is c 1 - c 6 alkyl or c 3 - c 8 cycloalkyl ; and , m and n are independently 0 , 1 , 2 or 3 . the invention also relates to pharmaceutically acceptable salts of these compounds and to pharmaceutically acceptable solvates thereof ; to compositions containing such compounds ; and to the uses of such compounds in the treatment of various diseases , particularly asthma and copd .

referring to fig1 a hand covering , in the form of a glove constructed in accordance with this invention is shown . the glove 10 includes an inner liner 12 and an outer shell 14 . the inner liner 12 is preferably formed of several layers of material . a preferred material for the inner layer 16 , i . e . that which immediately surrounds the hand , is a strong , high temperature resistant , cut , puncture and abrasion resistant fabric , such as one formed from aromatic polyamide fibers . the outer layer 18 of the inner liner 12 is preferably formed of a fabric made of a natural fiber such as cotton . one side of the material forming the outer layer 18 is knife coated with a light scrim coat , i . e . a very thin coat 20 , of a nitrile compound . the inner layer 16 is bonded to the non - coated side of the outer layer 18 by a laminator using an adhesive such as latex . palm and backhand portions of the glove are then cut from the laminated inner layer 16 and outer layer 18 , and secured to each other , such as by stitching or sonic welding , to form the inner liner 12 . thereafter a heavier coating 22 of a nitrile compound is provided on the outer surface of the outer layer 18 . in a preferred embodiment of this invention , the heavier coating of nitrile compound is applied to the outer layer 18 by dipping the liner 12 into a liquid bath of a nitrile compound one or more times . for instance , the liner 12 may be dipped into a container filled with a liquid nitrile compound as shown in fig2 . after curing , the nitrile compound provides a liquid proof coating or bladder 22 on the liner 12 . as an alternative to the nitrile compound , neoprene ® can be used . the neoprene ® when hardened forms a waterproof bladder over the natural fiber outer layer 18 . the neoprene ® coating or bladder has the desirable characteristic of being chemically resistant , so as to protect the hand from undesirable contact with fluids such as gasoline and hydraulic fluid . the outer shell 14 is made from a temperature - resistant aromatic polyamide fiber . the material which is to be used to form the palm portion of the outer shell 14 is provided with a kiss , or very thin coating 24 of silicone . a raised pattern 26 of silicone is applied over the thin coating 24 of silicone so as to provide both an enhanced gripping surface , and enhanced heat - resistance . while a particular raised pattern 26 of silicon is shown , other patterns may well be used with the same beneficial results . in a preferred embodiment of this invention , a two part silicone material is metered and mixed and then applied over the thin coating 24 of silicone through a nozzle as shown in fig3 . the nozzle is caused to move with respect to the palm portion of the outer shell 14 so as to form the desired raised pattern 26 of silicone which is thereafter cured . the movement of the nozzle with respect to the palm portion of the outer shell is in a preferred embodiment controlled by a computer . in applying the bead of silicone material to form the raised pattern 26 , it is desirable to space the bead away from the edges of the pattern for the palm portion , such that it will not interfere with the securing of the edges of the back and palm portions to each other , such as by sewing or sonic welding . in the preferred method of making the hand covering of this invention , the palm portion of the outer shell is cut from the material after the raised pattern 26 has been formed thereon . to complete the assembly of the glove , a wristlet 28 , also formed of a polyamide fabric is secured to the wrist ends of the inner liner 12 and the outer shell 14 . the wristlet is of a suitable length to protect the wrist and lower arm of the user . the wristlet 28 , which may be knit or formed as a woven cuff is preferably formed of a strong , high temperature resistant , cut , puncture and abrasion resistant fabric , such as one formed from aromatic polyamide fibers . in the preferred form of this invention , the inner liner is formed of 6 oz ./ sq . yd . 100 % kevlar ® inter - lock fabric . however , it is believed that kevlar ® and other aromatic polyamide fibers such as nomex ® aromatic polyamide fiber , manufactured by dupont and p . b . i .® polyamide fiber , manufactured by celanese , having a weight of 4 to 8 oz ./ sq . yd ., could be used in certain applications of this invention . the preferred fabric for the outer layer 18 of the inner liner 12 is 100 % cotton jersey fabric . cotton and other natural fiber fabrics having a weight of 4 to 7 oz ./ sq . yd . would be suitable for use in certain applications of this invention . as set forth above , the waterproof coating or bladder 22 on the liner 12 may be formed of a nitrile compound such as buna - n , synthetic rubbers or neoprene ®. however , other synthetic rubbers , such as butyl rubber , as well as polyurethane , may be used to form the waterproof bladder . while the preferred fabric for the outer shell is 9 oz ./ sq . yd . kevlar jersey fabric , fabrics formed from other aromatic polyamide fibers as mentioned above , having a weight of 8 to 10 oz ./ sq . yd . would be suitable for use in certain applications of this invention . it should be apparent to those skilled in the art , that while what has been described is considered at present to be the preferred embodiment of the protective hand covering of this invention , in accordance with the patent statutes , changes may be made in the hand covering without actually departing from the true spirit and scope of this invention . the appended claims are intended to cover all such changes and modifications which fall within the true spirit and scope of this invention .

a hand covering in the form of a glove which is water proof and provides protection against cutting , puncturing and lacerations as well as thermal insulation for protection against burning of the user &# 39 ; s hand when grasping hot objects . a raised silicone pattern is formed on the palm portion of the glove to enhance the heat insulating and gripping abilities of the hand covering .

referring to fig1 through 6 , one embodiment of this invention will be explained in the following . a floating body 1 is suspended on the liquid level 2 of liquid 3 , said liquid level 2 being kept in a predetermined height from the inside bottom 4 of the vessel 5 . the floating body 1 consists of a main body 6 and a plate member 7 . the main body 6 consists of a holding member 8 having rectangular opened fitting portions 9 , 9 , . . . in two lateral rows and culture ground members 10 , 10 , . . . fitted into each of said holes 9 , 9 , . . . the upper surface of the culture ground member 10 is used as a planting surface 10a which is kept to the position lower than the upper surface 8a of the holding member 8 . the lower surface 10b of the culture ground member 10 is kept to the position lower than the lower surface 8b of the holding member 8 . a holding member 8 has air ventilating holes 11 , 11 , . . . the culture ground member 10 is made of polyurethane foam and fed with water and nourishment . as shown in fig3 a plate member 7 consists of a plate portion 12 and a grid sheet 13 , said plate portion 12 having openings 14 , 14 , . . . which are the same as said opened fitting portions 9 , 9 , . . . with respect to shape and scale , said grid sheet 13 having a desired size of meshes 15 , namely a net having a number of fine rectangular meshes , which is adhered to the periphery of the opening 14 at the back surface of the plate portion 12 . the plate member 7 is provided with a perforated hole 16 at the position corresponding to air ventilating holes 11 , 11 , . . . of the holding member 8 . the lower surface 10b of the culture ground member 10 is positioned below the liquid surface 2 and a space 17 is provided between the liquid level 2 and the lower surface 8b of the holding member 8 . seeds 18 , 18 , . . . are planted on the planting surface 10a of the culture ground member 10 , 10 , . . . , the plate member 7 is put on an upper surface 8a of the holding member 8 , thus the opened fitting portion 9 is covered by the grid sheet 13 and also the space 19 of a predetermined distance is formed between the grid sheet 13 and the planting surface 10a of the culture ground member 10 . in this case , the plate member 7 should be mounted so as not to be separated from the upper surface 8a of the holding portion 8 . a supporting portion 20 of the holding member 8 supports the culture ground member 10 . water and nourishment are fed to planted seeds 18 on the planting surface 10a through the culture ground member 10 , because the lower surface 10b of the culture ground member 10 is positioned below the liquid level 2 . meanwhile , air is supplied to planted seeds 18 on the planting surface 10a through the culture ground member 10 , because the air is supplied to the space 17 through the holes 16 and the air ventilating holes 11 . seeds 18 put forth buds and take roots in which each stalk 21 is in the bending state and at the next growing stage the stalk 21 is still growing in the bending state with a seed coat 22 on its end which is in the position apart from the planting surface 10a . at further growing stage , a knee part 23 of the stalk 21 is caused to project through a mesh 15 of the grid sheet 13 , then the stalk 21 stretches itself at knee part 23 uprightly , at the same time the seed coat 22 is scraped by the crossbar portion of the mesh 15 of the grid sheet 13 due to the growing force of young seedling of welsh onion , and thus any seed coat 22 is no longer attached to the top end of the stalk 21 . each mesh 15 of the grid sheet 13 should be so designed as to prevent the passing of seed coat 22 ; namely it should be so designed that seed coat 22 can be scraped by the crossbar position of the mesh 15 when the stalk 21 stretches itself at the knee part 23 uprightly . when young seedlings of welsh onion are grown up to a predetermined length ( 10 cm to 25 cm ), they are put on the market in the state that they are planted on the culture ground member together with the plate member . fig7 and 8 show another embodiment of this invention , in which a culture ground member 24 is made of a sponge material and fitted into the vessel 25 . water and nourishment are fed into said culture ground member 24 through a perforated hole 26 provided at the bottom of the vessel 25 . seeds are planted on the planting surface 27 of the culture ground member 24 , a grid sheet 28 having a desired size of meshes 29 is adhered to the upper periphery of the opening 30 of the vessel 25 , and a tube shaped plastic film 31 at the lower end periphery thereof is adhered to the vessel 25 at the outer periphery of the opening 30 thereof . when young seedlings of welsh onion are grown up to a predetermined length they are put on the market in the state that they are planted in the culture ground member 24 . as mentioned above , this invention is to provide the method and apparatus for self - removing the seed coats which utilize the growing characteristic feature of young seedlings of welsh onion , and also which eliminate troublesome work for removing seed coats such as combing or cutting of the tops of young seedlings of welsh onion . thus , this invention can be applied to a mass production of young seedlings of welsh onion at high efficiency and has an effect for increasing its demand .

the present method and apparatus can be applied to cultivation of young seedlings of welsh onion economically in a mass production way utilizing the growing progresses of this vegetable , during which seed coats can be scraped and removed by the flange of each mesh of a screen sheet or screen film when a stalk stretches itself at the knee part uprightly . young seedlings of welsh onion are supplied as material for hors - d &# 39 ; oeuvre , soup and japanese &# 34 ; sushi &# 34 ;.

the present invention is directed toward a process utilizing enzymes to degrade and modify spent vanilla beans . typically , there may be used one or more enzymatic systems chosen from hydrolases ( e . c . 3 .) such as pectinase ( polygalacturonase ; ec 3 . 2 . 1 . 15 ), cellulases ( 1 , 4 -[ 1 , 3 ; 1 , 4 ]- β - d - glucan - 4 - glucano - hydrolase ; ec 3 . 2 . 1 . 4 ), β - glucosidases , proteases ( ec 3 . 4 . 23 ), hemicellulases ( glycan hydrolase e . c . 3 . 2 . 1 . ), xylanase ( 1 , 4 - β - d - xylanxylanohydrolase ; ec 3 . 2 . 1 . 8 ), lipases ( triacylglycerol lipase ec 3 . 1 . 1 . 3 ), esterases ( e . c . 3 . 1 . ), proteases ( e . c . 3 . 4 . ), oxidoreductases ( ec 1 . 1 .- ec 1 . 11 ), and transferases ( e . c . 2 .). these enzymes may be used alone or in a mixture with β - glucosidase . these enzymes are well known to persons skilled in the art and many systems based thereon are commercially available . the broader implication of this discovery includes the economical production of products and intermediary by - products . it is understood that the invention covers all the enzymatic systems , which make it possible to degrade and transform the precursors potentially contained in the spent ( exhausted ) vanilla pods . persons skilled in the art will be able , using simple procedures such as those defined in the examples which follow , to choose the enzymatic systems that are appropriate . preferably , the spent vanilla pods are treated with an enzymatic system which comprises : at least one enzyme capable of destroying the cell membrane systems of plant cells and is selected from the group consisting of an enzyme having hydrolase activity , such as pectinase activity , an enzyme having cellulase activity , an enzyme having hemicellulase activity , an enzyme having protease activity , an enzyme having lipase activity , and an enzyme having β - glucosidase activity , an enzyme having oxidoreductase activity or an enzyme having transferase activity . the enzyme is utilized in an amount of about 10 to about 1000 units per gram of spent vanilla pods . it is further understood that the spent vanilla beans are ground and hydrated in water . generally , without being an essential feature of the present invention , spent vanilla pods are present in the amount of between 5 to 50 % of the water added . the enzymatic system advantageously comprises 10 to 1000 units of enzyme activity per gram of spent vanilla pods , and preferably from 20 to 500 units of enzymatic activity . it was observed that a range of between 40 and 400 units of enzymatic activity was even more advantageous for implementing the process according to the invention . the enzymatic reaction is carried out at a ph which is advantageously between about 3 and 10 , and preferably at the optimum ph for each individual enzyme system . as the ph of the ground , spent vanilla pods product obtained is approximately equal to 5 , this ground product is therefore naturally at the optimal value for enzymes such as cellulases , pectinases , hemicellulases , and β - glucosidases . enzyme systems containing lipases , proteases , esterases , oxidoreductases and transferases require a ph that is optimal for each enzyme system . the process is carried out with stirring for a period of time sufficient to allow the degradation and modification of the spent vanilla pods and the release of the natural flavor components . advantageously , this period will be greater than 2 hours at room temperature . this processing temperature can be increased or decreased while being careful not to exceed a maximum temperature which can result in degradation of the flavor precursors and enzymes . decrease of the temperature too substantially can cause slowing or cessation of the desired reactions . the temperature will be generally between about 10 and 100 degrees c ., preferably between about 30 and 40 degrees c . regarding the time that incubation takes place , the period of incubation will be between about 2 and 120 hours . however , it has been observed that the release is generally complete after a few hours of incubation , it being possible for persons skilled in the art to determine substantial completion by means of high performance liquid chromatography ( hplc ) or gas chromatography mass spectroscopy ( gcms ) analyses . after incubation , the liquid phase containing the vanilla flavor is separated from the solid phase , which contains especially insoluble cell residues . this separation may be carried out , for example , by filtration and / or by centrifugation . the liquid phase containing the natural flavor compounds and food modifiers may then be used , either directly or after concentration of the compounds . such concentration may be carried out by evaporation , selective chromatography , optionally under vacuum , and then filtration . it may also be carried out by extraction with solvents and subsequent evaporation of the latter . the following examples are intended to illustrate the invention without limiting it . this example demonstrates the effect of different enzymes on the release of flavor components and their sensory attributes . ten grams ( 10 g ) of dry , spent vanilla pods were ground and mixed with 90 g of deionized water . a slurry consisting of two phases , solid and liquid , was stirred . different enzymes such as cellulase , xylanase , hemicellulase , pectinase , protease , lipases and oxidoreductase were typically added in the amount of 500 - 1 , 000 units per gram of dry spent vanilla pods . these enzymes were applied either alone or in combination with each other . one unit of cellulase liberates 1 . 0 μmol of reducing sugar measured as glucose from cellulose in one hour at a ph 5 at 37 degree c . one unit of xylanase liberates 1 . 0 μmol of reducing sugar measured as xylose equivalents from xylan per minute at ph a 4 . 5 at 30 degree c . one unit of hemicellulase liberates 1 . 0 μmol of d - galactose from hemicellulose per hour at a ph 5 . 5 at 37 degree c . one unit of pectinase liberates 1 . 0 μmol of galacturonic acid from polygalacturonic acid per minute at a ph 4 . 0 at 25 degree c . one unit of protease hydrolyzes hemoglobin to produce color equivalent to 1 . 0 μmol of tyrosine per minute at a ph 2 . 8 at 37 degree c . one unit of lipase hydrolyzes 1 . 0 microequivalent of fatty acid from a triglyceride in 1 hour at ph 7 . 7 at 37 degree c . activity of laccase is expressed in nanokatal / ml ; one nanokatal is the amount of enzyme needed to perform oxidative dimerization of one nanomole of 2 , 6 - dimethoxyphenol ( slomczynski et al ., 1995 ). the incubation with the enzymes was typically carried out without ph adjustment at the natural ph of around 5 . 0 . the ph was measured after addition of the enzyme and at the end of the enzymatic reaction and recorded . the slurry was mixed while mixture was incubated at 35 degree c ., typically for 10 hours . at the end of the reaction , the mixture is filtered rapidly through a whatman filter paper . a small portion ( 10 ml ) of the filtrate was extracted with 5 ml of methylene chloride . the methylene chloride phase was concentrated and subjected to gc / ms analysis . the remaining filtrate was mixed with 96 % alcohol to obtain a 50 % aqueous - alcoholic medium . the filtrate was evaluated by sensory panel . the hydrolysis released a number of compounds from spent vanilla beans . table 1 shows only the level of vanillin released and compares the sensory attributes of the samples . ten grams ( 10 g ) of dry , spent vanilla pods were ground and mixed with 90 g of deionized water . a slurry consisting of two phases , solid and liquid , was formed and agitated . the enzyme , β - glucosidase , was added in the amount of 53 units of enzyme per gram of dry spent vanilla pods . one unit of enzyme activity is defined as the amount of enzyme which would liberate 1 . 0 μg of p - nitrophenol from p - nitrophenyl - glucoside at 35 degrees c . at ph 5 . the incubation of spent vanilla pods with the enzyme β - glucosidase was carried out without ph adjustment at the natural ph of about 5 . 0 . the ph was measured after addition of the enzyme and at the end of the enzymatic reaction and recorded . the slurry was mixed while incubated at 35 degree c . typically for 12 hours . at the end of the reaction , the solid material was quickly filtered through a whatman filter paper . a small portion ( 10 ml ) of the filtrate was extracted with 5 ml of methylene chloride . the methylene chloride fraction was concentrated and subjected to gc / ms analysis . the remaining liquid filtrate was mixed with 96 % alcohol to obtain a 50 % aqueous - alcoholic flavor mixture . this filtrate was evaluated by a sensory panel . the hydrolysis released a number of compounds from the spent vanilla beans . table 2 shows only the level of vanillin released and defines the sensory attributes of the sample . ten grams ( 10 g ) of dry , spent vanilla pods were ground and mixed with 90 g of deionized water . a slurry consisting of two phases , solid and liquid , was stirred . two enzymes , cellulase and β - glucosidase were added in the amount of 1 , 000 and 100 units per gram of dry spent vanilla pods , respectively . the incubation with the enzymes was typically carried out without ph adjustment at the natural ph of around 5 . 0 . the ph was measured after addition of the enzyme and at the end of the enzymatic reaction and recorded . the slurry was mixed while incubated at 35 degree c ., typically for 10 hours . at the end of the reaction , the mixture was mixed with 96 % ethyl alcohol to obtain 50 % aqueous - alcohol solution . the mixture was mixed for 3 hours at room temperature and filtered through a whatman filter paper . the filtrate was evaluated by a sensory panel . the essence of the present invention is a process for obtaining a natural flavor extract and a food modifier from the hydrated product of spent vanilla pods by treating the pods in an enzymatic system capable of degrading the plant cells and / or modifying precursors in spent vanilla beans . ground , spent vanilla beans are used to facilitate the enzymatic catalysis of the cell components . finer ground products will allow the enzymes to perform the reaction faster . spent vanilla pods are understood to mean cured vanilla pods from which the vanilla components have already been extracted , typically with ethanol or carbon dioxide . an enzymatic system capable of disintegration and modification of the spent vanilla bean cells is understood to mean all the systems which can liquefy fruits , vegetables and , in general , all plant cells . it also means enzymes which can modify and transform existing chemical entities present in the spent vanilla . it will be apparent to those of skill in this art that the foregoing examples are only illustrative of my invention and those modifications thereof and additions thereto will be apparent to those of skill in this art . as to all such modifications and additions , it is desired that they be included within the purview of my invention , which is to be limited only by the scope , including equivalents , of the following , appended claims

a process for obtaining flavor components from spent vanilla pods and beans , comprises forming a mixture of such spent pods and beans , incubating the mixture with an enzyme capable of destroying the cell membrane systems of said pods and beans for period of time and at a ph and temperature such that said cell membranes break down and release flavor constituents , and thereafter recovering the flavor constituents from the mixture .

while this invention is susceptible of embodiments in many different forms , there will herein be described in detail a preferred embodiment of the invention with the understanding that the present disclosure is to be considered as an exemplification of the principles of the invention and is not intended to limit the broad aspect of the invention to any specific embodiments . although the exact cause or causes for canker sores are not well - known , a large population has suffered from this potentially painful condition . the flair up of a canker sore can make it painful for a person to speak , drink , swallow or eat , depending the location and size of the canker sores . for many individuals , repeat occurrences are frequent and unpredictable . various likely causes of canker sores have been proposed . a comparison of the more popular proposed causes points to a few commonly observed factors . one likely factor is a deficiency in the intake of vitamins , including b 6 , b 9 and b 12 , and minerals such as zinc . another major factor that has been frequently cited as a cause of canker sores is stress . still another cited factor for canker sore formation is trauma to the mouth , such as an accidental bite of the soft cheek tissue or the tongue . it is also believed that acidity can exacerbate the sores , which is why many medical professionals suggest canker sore sufferers avoid highly acidic food and drinks . while none of these factors alone may cause canker sores , the more factors present increases the likelihood of canker sore formation . the present formulation is directed to an ingestible mouth spray that aims at inhibiting the development of canker sores at an early stage and at healing existing canker sores should they develop . the preferred composition is constituted totally of non - toxic chemical ingredients . generally speaking , the formulation includes the use of an alkaline solution as an effective treatment of canker sores . the preferred formula also contains at least three b vitamins , and zinc , as deficiencies in these vitamins and mineral are commonly - known hosting factors of canker sores . most preferably , the b vitamins include vitamin b 6 , b 9 , and b 12 , but may include additional b vitamins . a preferred embodiment of the formula contains the following chemical ingredients and measures ( in parts - per - million ( ppm )) in an aqueous solution : 1 . carbonate : 1200 ppm in an alkali metal salt 2 . vitamin b 6 : 3 . 0 ppm 3 . vitamin b 9 : 1 . 0 ppm 4 . vitamin b 12 : 5 . 0 ppm 5 . zinc : 65 ppm in zinc ( ii ) edta alkali metal salt 6 . methycarboxycellulose : 1000 ppm in methycarboxycellulose alkali metal salt , as a thickening agent 7 . a natural flavoring chemical ( such as peppermint ): very minute amount , as needed preferably , the alkali metal is sodium ( na 30 ), but may also be potassium ( k + ), lithium ( li + ), or cesium ( cs + ). further , the formula can be modified and the following ranges for the chemical ingredients are possible for alternate embodiments in an aqueous solution : 1 . carbonate : 12 to 12000 ppm in an alkali metal salt 2 . hydroxide : 17 to 17000 ppm in an alkali metal salt 3 . vitamin b 6 : 0 . 0 to 5 . 0 ppm 4 . vitamin b 9 : 0 . 0 to 5 . 0 ppm 5 . vitamin b 12 : 0 . 0 to 15 . 0 ppm 6 . other b vitamins : 0 . 0 to 5 . 0 ppm 17 . zinc : 0 . 1 to 150 ppm in zinc ( ii ) edta alkali metal salt 8 . methylcarboxycellulose : 30 - 3000 ppm in alkali metal salt , as a thickening agent 9 . a natural flavoring chemical ( such as peppermint ): 0 . 0 to as needed due to the non - toxicity of the composition , it can be applied to and held in the mouth as often and as long as the user prefers . this in turn allows longer contact between the mucosal membrane of the user &# 39 ; s mouth and the composition . a user could allow an optimal time for an effective chemical reaction between the ingredients and the tissue , including any canker sores . afterward , the composition can be swallowed . this provides a significant advantage over non - ingestible treatments , such as mouthwashes — i . e ., it is not necessary to spit it out . the fact that this product is ingestible greatly enhances the timing that this product can be conveniently used or applied — the user can carry the product in his / her handbag or briefcase and use it almost anytime and almost anywhere . no longer does treatment require a bathroom sink or the like . the product can be administered at the user &# 39 ; s home while watching tv , at the user &# 39 ; s office desk , in the airport without toting bags to the restroom , at the bus stop and on the bus , in a restaurant , and in an almost infinite number of other locations where it would be otherwise be inappropriate to dispel a mouthwash . more frequent and longer applications of the treatment lead to more effective prevention and treatment of canker sores . further , the chemical composition results in a product which is neutral in both taste and smell . as such , the use of the composition , even in intimate social gatherings , is not offensive to others and significantly reduces the occasions for missing a treatment . in a preferred formulation , vitamin b 6 ( pyridoxine , pyridoxal , or pyridoxamine , or pyridoxine hydrochloride ), b 9 ( folic acid ), b 12 ( various cobalamins ; commonly cyanocobalamin in vitamin supplements ) and zinc have been added to the alkaline solution to improve the intake of these components and reduce the cause of canker sores due to the deficiencies in these components . other b vitamins , such as b 1 ( thiamine ), b 2 ( riboflavin ), b 3 ( niacin or niacinamide ), b 5 ( pantothenic acid ) and b 7 ( biotin ; vitamin h ) may be substituted or added to other formulations , if desired . the inclusion of zinc in a preferred formulation is due to the fact that , in scientific literature , zinc has also been shown to be anti - ulceration and thus , will further help to reduce the likelihood of forming canker sores ( ulceration of the mucous of the mouth ). mental stress is believed to be a major and probably the most frequent contributing factor for the occurrence of canker sores . as much as stress is a psychological condition , it is however extremely import to realize that , this psychological condition does produce physiological changes . in 2012 , proceedings of national academy of sciences , usa , released a report that correlates stress and tissue inflammations . this strongly supports the long clinical observation that stress results in tissue inflammation . inflammation biochemically results in acidosis of the body &# 39 ; s tissues . there have been many studies which support the fact that under stress , many tissues / organs are more acidic than those of a control group . the ulcerations of the tissues come as a result of the prolonged stress and prolonged acidosis of the tissues , such as the mucous membranes of the mouth . trauma to the mouth , such as an accidental bite of the soft cheek or the tongue is believed to be another factor for the occurrences of canker sores . an unhealed injury to any tissues is known to incite the inflammation reactions from the body . thus , just like mental stress , trauma to the mouth also results in the inflammations and acidosis of the injured tissues in the mouth , resulting in the formation of canker sores . many medical professionals suggest canker sore sufferers avoid highly acidic food and drinks . this clinical advice correlates well with the fact that canker sores , irrespective of the causes , are in principle results of the inflammation and the ensuing acidosis of the mucous tissues and extracellular acidity from the drinks or foods will only exacerbate the inflammation . the disclosed formulation , therefore , directs to reverse the acidosis — and the possible ensuing ulcerations — of the mucous membranes of the mouth during early phases in the development of canker sores . such treatment , as described , will result in prevention of and quicker healing of canker sores . with the inclusion of methylcarboxycellulose alkali metal salt , as a thickening agent , the preferred formulation is considered to be stable ( for shelf - life consideration ) and is a clear , non - toxic and ingestible alkaline solution . it is also very neutral and can be pleasant in taste , especially when a natural flavoring chemical is included . and , it has no offensive odor to others . the present embodiment can thus be applied in any work or social occasion without missing a dose . the matter set forth in the foregoing description is offered by way of illustration only and not as a limitation . while particular embodiments have been described , it will be apparent to those skilled in the art that changes and modifications may be made without departing from the broader aspects of applicants &# 39 ; contribution . the actual scope of the protection sought is intended to be defined in the following claims when viewed in their proper perspective based on relevant prior art .

a treatment and prevention formulation for canker sores of the oral cavity including an aqueous alkaline solution having at least three b vitamins and zinc edta alkali metal salt ) mixed therein and administered to a target area by a spray container . the vitamins are b 6 , b 9 and b 12 , but may include other vitamins as well . the formulation is totally non - toxic in chemical nature and non - offensive in odor to others , so it can be used anywhere and at any time and can be swallowed after use .

in a preferred embodiment , the water insoluble thermoplastic polymer is either cellulose or neutral ester copolymer ( e . g ., neutral methacrylic acid esters such as eudragit ® ne , eudragite ® rl , and eudragit ® rs ) or mixture thereof . in still another preferred embodiment , the loading dose is provided by the same or different therapeutic agent ( s ) in the central drug compartment . in yet another preferred embodiment there is present one or more surface active agents and as an example glyceryl monooleate , sodium lauryl sulphate or glyceryl monostearate . in a preferred embodiment , release of therapeutic agent is characterized by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within two hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within three hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within fours hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within five hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within six hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within eight hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within twelve hours , and greater than 50 % is released in 24 hours , or by a dissolution profile in which at least 0 . 5 % to 50 % of the therapeutic agent is released within sixteen hours , and greater than 50 % is released in 24 hours the oils used in the invention can be one or more selected from almond oil , apricot kernel oil , avocado oil , black currant oil , 14 % gla , borage oil , 20 % gla , canola oil , carrot oil , castor oil , clove leaf oil , coconut oil , corn oil , cottonseed oil , evening primrose oil , 9 % gla , flaxseed oil , 55 % ala , grapeseed oil , hazelnut oil , hemp oil , ala / gla , hydrogenated oils , jojoba oil , golden jojoba oil , water - white kukui nut oil , macadamia nut oil , oat oil , olive oil , extra virgin olive oil pomace /“ b ” grade , olive oil , pure / nf , palm oil , parsley seed oil , peach kernel oil , peanut oil , pecan oil , pistachio oil , pumpkinseed oil , rice bran oil , rose hip seed oil , rosemary oil , safflower oil , linoleic &# 39 ; safflower oil , high - oleic , sesame oil nf , sesame oil toasted , soybean oil , sunflower oil , salad sunflower oil high - oleic , tea tree oil , vegetable , glycerine , usp , walnut oil , wheat germ oil , cold - pressed and mineral oil or other similar oils . the gas generators used in the invention can be selected from soluble or insoluble acid sources such as food acids ( citric acid , tartaric acid , lactic acid , malic acid , fumaric acid , ascorbic acid , adipic acid , succinic acid ); acid anhydrides ( succinic anhydride , citric anhydride ); acid salts ( sodium dihydrogen phosphate , disodium dihydrogen pyrophosphate , sodium dihydrogen citrate , disodium hydrogen citrate ) and carbonate sources ( sodium bicarbonate , sodium carbonate , potassium bicarbonate , potassium carbonate , sodium sesquicarbonate , sodium glycine carbonate , lysine carbonate , arginine carbonate , amorphous calcium carbonate ); and oxygen gas generators such as anhydrous sodium perborate or other suitable generators the antioxidants used in this invention may be selected from ascorbic acid , fumaric acid , malic acid , alpha tocopherol , ascorbic acid palmitate , butylated hydroxyanisole , propyl gallate , sodium ascobate , and sodium metabisulfite or other suitable antioxidants the said invention also accomplishes these and other objectives through a method for preparing a tablet , pellet or bead for use in controlled extended release of therapeutic agent in which dry or wet granulation of an effective amount of therapeutic active agent , or a pharmaceutically acceptable salt thereof , and dry or wet granulating aids , tableting aids and gas generators and optionally oil is carried out using high shear or low shear mixers or fluidbed granulators . if required , the granules are dried , sieved , lubricated , blended before being compressed to form a tablet or pellet to form a central compartment . this is followed by coating using fluid bed or pan coating techniques to apply one or more coats of thermoplastic cellulose ether in combination with an acrylic polymer and or cellulose esters and superdisintegrant and optionally oil and electrolytes . additionally , a top coat may be added for the purpose of delivering a loading dose and or serving as a timing device made from acrylic polymers or cellulose esters . a preferred embodiment further includes the steps of combining an effective amount of a therapeutic active agent , or a pharmaceutically acceptable salt thereof , gas generators , optionally oil , and extrusion spheronization aids to form beads or pellets made from extrusion spheronization techniques . the beads may also be made by drug powder or solution layering . the beads formed are coated using fluid bed or pan coating to apply one or more coats of thermoplastic cellulose ether in combination with an acrylic polymer and or cellulose esters and superdisintegrant and optionally oil and electrolytes and antioxidants . additionally a top coat may be added for the purpose of serving as a loading dose and or serving as a timing device made from acrylic polymers or cellulose esters . in a preferred embodiment , a pharmaceutically acceptable acrylic polymer , includes , but is not limited to , acrylic acid and methacrylic acid copolymers , methyl methacrylate copolymers , ethoxyethyl methacrylates , cyanoethyl methacrylate , aminoalkyl methacrylate copolymer , poly ( acrylic acid ), poly ( methacrylic acid ), methacrylic acid alkylamide copolyer , poly ( methyl methacrylate ), poly ( methyl methacrylate ) copolymer , polyacrylamide , aminoalkyl methacrylate copolymer , poly ( methacrylic acid anhydride ), and glycidyl methacrylate copolymers . additionally , the acrylic polymers may be cationic , anionic , or non - ionic polymers and may be acrylates , methacrylates , formed of methacrylic acid or methacrylic acid esters . the polymers may also be ph independent or ph dependent . it is to be understood that more than one therapeutically active agent may be incorporated into the device of this invention , and that the use of the term “ agent ” or “ drug ” in no way excludes the use of two or more such agents or drugs . the therapeutic agents can be in various forms , such as uncharged molecules , components of molecular complexes or nonirritating , pharmacologically acceptable salts . also , simple derivatives of the agents ( such as ethers , esters , amides , etc .) which are easily hydrolyzed by body ph , enzymes , etc ., can be employed . the term “ therapeutically active agent ”, or “ therapeutic agent ”, or “ active agent ” as used herein refers to an agent , drug , compound , composition of matter or mixture thereof which provides some biological , often beneficial , effect . this includes pesticides , herbicides , germicides , biocides , algaecides , rodenticides , fungicides , insecticides , antioxidants , plant growth promoters , plant growth inhibitors , preservatives , antipreservatives , disinfectants , sterilization agents , catalysts , chemical reactants , fermentation agents , foods , food supplements , nutrients , cosmetics , drugs , vitamins , sex sterilants , fertility inhibitors , fertility promoters , microorganism attenuators and other agents that benefit the environment of use . as used herein , the terms further include any physiologically or pharmacologically active substance that produces a localized or systemic effect or effects in animals , including warm blooded mammals , humans and primates ; avians ; domestic household or farm animals such as cats , dogs , sheep , goats , cattle , horses and pigs ; laboratory animals such as mice , rats and guinea pigs ; fish ; reptiles ; zoo and wild animals ; and the like . the active agent that can be delivered includes inorganic and organic compounds , including , without limitation , drugs which act on the peripheral nerves , adrenergic receptors , cholinergic receptors , the skeletal muscles , the cardiovascular system , smooth muscles , the blood circulatory system , synoptic sites , neuroeffector junctional sites , endocrine and hormone systems , the immunological system , the reproductive system , the skeletal system , autacoid systems , the alimentary and excretory systems , the histamine system and the central nervous system . suitable active agents may be selected from , for example , proteins , enzymes , hormones , polynucleotides , nucleoproteins , polysaccharides , glycoproteins , lipoproteins , polypeptides , steroids , hypnotics and sedatives , psychic energizers , tranquilizers , anticonvulsants , muscle relaxants , antiparkinson agents , analgesics , anti - inflammatories , local anesthetics , muscle contractants , antimicrobials , antimalarials , hormonal agents including contraceptives , sympathomimetics , polypeptides and proteins capable of eliciting physiological effects , diuretics , lipid regulating agents , antiandrogenic agents , antiparasitics , neoplastics , antineoplastics , hypoglycemics , nutritional agents and supplements , growth supplements , fats , ophthalmics , antienteritis agents , electrolytes and diagnostic agents . examples of beneficial active agents useful in this invention include prochlorperazine edisylate , ferrous sulfate , aminocaproic acid , mecaxylamine hydrochloride , procainamide hydrochloride , amphetamine sulfate , methamphetamine hydrochloride , benzphetamine hydrochloride , isoproteronol sulfate , phenmetrazine hydrochloride , bethanechol chloride , methacholine chloride , pilocarpine hydrochloride , atropine sulfate , scopolamine bromide , isopropamide iodide , tridihexethyl chloride , phenformin hydrochloride , methylphenidate hydrochloride , theophylline cholinate , cephalexin hydrochloride , diphenidol , meclizine hydrochloride , prochlorperazine maleate , phenoxybenzamine , thiethylperazine maleate , anisindione , diphenadione erythrityl tetranitrate , digoxin , isoflurophate , acetazolamide , methazolamide , bendroflumethiazide , chlorpropamide , tolazamide , chlormadinone acetate , phenaglycodol , allopurinol , aluminum aspirin , methotrexate , acetyl sulfisoxazole , hydrocortisone , hydrocorticosterone acetate , cortisone acetate , dexamethasone and its derivatives such as betamethasone , triamcinolone , methyltestosterone , 17 -. beta .- estradiol , ethinyl estradiol , ethinyl estradiol 3 - methyl ether , prednisolone , 17 -. beta .- hydroxyprogesterone acetate , 19 - nor - progesterone , norgestrel , norethindrone , norethisterone , norethiederone , progesterone , norgesterone , norethynodrel , aspirin , indomethacin , naproxen , fenoprofen , sulindac , indoprofen , nitroglycerin , isosorbide dinitrate , propranolol , timolol , atenolol , alprenolol , cimetidine , clonidine , imipramine , levodopa , chlorpromazine , methyidopa , dihydroxyphenylalanine , theophylline , calcium gluconate , ketoprofen , ibuprofen , cephalexin , erythromycin , haloperidol , zomepirac , ferrous lactate , vincamine , phenoxybenzamine , milrinone , captropril , mandol , quanbenz , hydrochlorothiazide , ranitidine , flurbiprofen , fenbufen , fluprofen , tolmetin , alclofenac , mefenamic , flufenamic , difuninal , nimodipine , nitrendipine , nisoldipine , nicardipine , felodipine , lidoflazine , tiapamil , gallopamil , amlodipine , mioflazine , lisinopril , enalapril , captoril , ramipril , enalaprilat , famotidine , nizatidine , sucralfate , etintidine , tetratolol , minoxidil , chlordiazepoxide , diazepam , amitriptylin , and imipramine . further examples are proteins and peptides which include , but are not limited to , insulin , colchicine , glucagon , thyroid stimulating hormone , parathyroid and pituitary hormones , calcitonin , renin , prolactin , corticotrophin , thyrotropic hormone , follicle stimulating hormone , chronic gonadotropin , gonadotropin releasing hormone , bovine somatotropin , porcine somatropin , oxytocin , vasopressin , prolactin , somatostatin , lypressin , pancreozymin , luteinizing hormone , lhrh , interferons , interleukins , growth hormones such as human growth hormone , bovine growth hormone and porcine growth hormone , fertility inhibitors such as the prostaglandins , fertility promoters , growth factors , and human pancreas hormone releasing factor . as used herein , the terms “ therapeutically effective ” amount or rate refer to the amount or rate of the active agent needed to achieve the desired therapeutic result . the following examples are illustrative only , and not limiting of the remainder of the disclosure in any way whatsoever . formulation 1 , 2 or 3 may be used to make tablets or pellets . carvedilol is mixed with microcystalline cellulose , tartaric acid and or citric acid , calcium carbonate or sodium bicarbonate , oil , glyceryl monooleate , sodium lauryl sulphate , polysorbate 80 and silicon dioxide in a high - shear or low shear mixer granulator or patterson - kelley v - blender for 10 minutes . the mixture is then granulated with an organic solvent and dried . granulation may be done in a fluidbed . magnesium stearate is added to the dried granules and blended for 5 more minutes in a v - blender . the powder blend is then compressed using an elizabeth hata rotary tablet press . formulation 4 or 5 may be used to make beads . carvedilol , microcrystalline cellulose , citric acid , sodium bicarbonate , sodium lauryl sulphate , polysorbate 80 , glyceryl monooleate and oil are dry mixed in a low shear mixer and wet granulated . the wet mass is extruded and spheronized using a caleva extruder and spheronizer . the spheronized beads are dried . ethylcellulose is dissolved in alcoholic solution . to this is added an acrylic polymer ( eugragit l and eudragit s ), super disintegrant ( crospovidone ) and oil ( coconut oil ). this is sprayed on the tablets ( using a side vented pan coater ), pellets or beads ( using a fluid bed coater ) to form a coat or coats around them to a weight gain of from about 1 . 0 % to about 25 %. formulation 1 , 2 or 3 may be used to make tablets or pellets . venlafaxine is mixed with microcystalline cellulose , tartaric acid and or citric acid , calcium carbonate or sodium bicarbonate , oil , polysorbate 80 and silicon dioxide in a high - shear or low shear mixer granulator or pafterson - kelley v - blender for 10 minutes . the mixture is then granulated with an organic solvent and dried . granulation may be done in a fluidbed . magnesium stearate is added to the dried granules and blended for 5 more minutes in a v - blender . the powder blend is then compressed using an elizabeth hata rotary tablet press . formulation 4 or 5 may be used to make beads . venlafaxine , microcrystalline cellulose , citric acid , sodium bicarbonate , polysorbate 80 , and oil are dry mixed in a low shear mixer and wet granulated . the wet mass is extruded and spheronized using a caleva extruder and spheronizer . the spheronized beads are dried . ethylcellulose is dissolved in alcoholic solution . to this is added hydroxypropylmethyl cellulose acetate , super disintegrant and sodium chloride . this is sprayed on the tablets , pellets or beads to form a coat or coats around them to a weight gain of from about 1 . 0 % to about 25 %. ( v ) composition and addition of loading dose of venlafaxine 5 % wt by wt . a loading dose consisting of venlafaxine is applied as a coat to a weight gain of about 5 %. this consists of venlafaxine in a hydroxypropylmethyl cellulose binder solution . the coat is made from eudragit l and s in ratio of 5 : 1 . and is applied to a coating thickness of from about 1 % to about 50 % weight gain this is the same as example 2 except that venlafaxine is replaced by paroxetine hydrochloride and sodium chloride is added to the ethylcellulose coat . this is the same as example 2 except that venlafaxine is replaced by metoprolol succinate and sodium chloride and coconut oil are added to ethylcellulose coat this is the same as example 2 except that venlafaxine is replaced by metoprolol succinate and ascorbic acid , sodium chloride and coconut oil are added to the cellulose coat .

a controlled extended drug release technology for the controlled extended release of hydrophobic or hydrophilic drugs or therapeutically active agents consisting of a homogeneous blend of one or more therapeutic agents , gas generators and surrounded by one or more layers of coat made of thermoplastic water insoluble cellulose derivatives , acrylic polymers , superdisintegrants and optionally an oil , antioxidants and electrolytes . the technology platform is capable of releasing therapeutic agents via zero , first or pseudo first order release .

turning now to the drawings , in fig1 , a cotton harvester 10 is shown , including an on - board cotton packager or module builder 12 for compacting cotton harvested by harvester 10 into a unitary cotton module ( not shown ). referring also to fig2 , a compactor 14 operable according to the present invention is shown . compactor 14 includes a compactor frame 16 which is oriented generally horizontally , or within a range of small acute angles relative to horizontal , and substantially entirely disposed within a compactor chamber 18 , for movement downwardly against cotton contained therein for compacting the cotton against a floor 20 therein . compactor frame 16 includes a front cross member 22 disposed in chamber 18 adjacent a front wall 24 , and having opposite ends which extends through sidewardly open slots 26 in the sides of module builder 12 . similarly , a rear cross member 28 is disposed in chamber 18 and has opposite end portions which extend through slots 30 in sides of module builder 12 . augers 32 are supported in forward and rearward extending relation between cross members 22 and 28 within chamber 18 . augers 32 can be rotated using any suitable commercially available drivers , such as a gear drive driven by a motor such as a fluid or electric motor , or directly by fluid or electric motors , as desired , and as controlled by an auger on solenoid 34 and an auger reverse solenoid 36 ( fig3 ), for distributing the collected cotton in chamber 18 as will be explained . in this regard , it should be noted that it is desirable and a sought after feature to distribute the cotton evenly with respect to the plane of floor 20 , such that the resultant compacted cotton module will have a substantially uniform height along its length and width . compactor frame 16 of compactor 14 is supported in compacting chamber 18 on each side by an exterior side structure 38 , each structure 38 including a forwardly and rearwardly extending main beam 40 which extends between and connects front and rear cross members 22 and 28 . each side structure 38 additionally includes a pair of braces 42 which extend downwardly and at converging angles from front and rear cross members 22 and 28 , and which are connected together by a gusset 44 located spacedly below about the middle of main beam 40 . here , it should be noted that compactor frame 16 located within compacting chamber 18 and exterior side structures 38 on the exterior of module builder 12 are movable upwardly and downwardly together . the upward and downward movement of exterior side structures 38 and compactor frame 16 is preferably achieved and controlled by fluid cylinders 46 extending , respectively , between gussets 44 of each exterior side structure 38 and a support frame 48 supported by and extending upwardly from a frame 50 of module builder 12 . importantly , a rod 52 of each cylinder 46 is connected to gusset 44 at a pivot 54 which allows limited pivotal movement of side structure 38 and thus compactor frame 16 and augers 32 of compactor 14 about a side - to - side extending pivotal axis within a limited range of pivotal movement . support frame 48 on each side of module builder 12 includes a pair of diagonally extending braces 56 having lower ends connected to frame 50 , and upper ends which connect to and support vertical braces 58 which support a cross member 60 to which fluid cylinder 46 is attached . a more forward brace 56 of support frame 48 on that side of module builder 12 facing outwardly from the page , and the more rearwardly located brace 56 on the opposite side of the module builder , support forward and rear compactor position sensors 62 a and 62 b , respectively . each compactor position sensor 62 a and 62 b includes an elongate actuator arm 64 which pivotally connects to gusset 44 on that side of the module builder . each sensor 62 a and 62 b is a rotary type sensor , which will detect rotational movement of the respective actuator arm 64 , as compactor 14 is moved from the position shown in fig1 , for instance , to any of several lowered positions ( not shown ). because two compactor position sensors 62 a and 62 b are used , movements of compactor 14 at a tilt will result in different rotational displacements of actuator arms 64 of the respective sensors 62 a and 62 b , and thus the sensors will output different positional values . the difference between these positional values can be utilized for determining both the vertical position of compactor 14 , and also any tilt thereof . compactor position sensors can include , for instance , potentiometers , which vary a voltage or current signal when an input thereof is rotated . actuator arms 64 can be slidable relative to the input to prevent binding when rotated . for instance , a vertical position of the compactor can be determined as a function of a sum of the values outputted by sensors 62 a and 62 b , such as by averaging the values . referring also to fig3 , a compactor controller 66 of packager 12 is operable for receiving signals outputted by a number of devices , including , but not limited to , a compactor pressure signal from a compactor pressure sensor 68 , two compactor position signals outputted by compactor position sensors 62 a and 62 b , auger pressure signals outputted by an auger pressure sensor 70 , and one or more additional signals outputted by various sensors or other devices . controller 66 can be connected to the sensors using any suitable conductive paths , such as , but not limited to , a conventional wiring harness , optical cables , a wireless network , or the like . responsive to the signals from these devices , and / or other devices , controller 66 is operable for automatically responsively outputting signals to apparatus such as a compactor raise solenoid 72 and a compactor lower solenoid 74 , which control fluid cylinders 46 ( fig1 ) operable for moving compactor 14 upwardly and downwardly against cotton accumulated in a bottom region of compactor chamber 18 . the cylinders 46 can also be used for setting or indexing the compactor position . compactor controller 66 is also operable for outputting signals to augers 32 ( fig1 and 2 ), for effecting forward or reverse rotation thereof via auger on solenoid 34 and auger reverse solenoid 36 , and to other suitable devices . as indicated above , compactor controller 66 is preferably programmed for automatically controlling compactor and auger operation for building a compacted cotton module within compactor chamber 18 , as a function of the outputs from the sensors , time , and other parameters , as harvester 10 is operating . however , failure or fault of a sensor or sensors , and / or of the conductive path connecting a sensor or sensors to controller 66 , either of a continuous , intermittent , or erratic manner , could result in a lack of complete data required for normal automatic programmed operation of compactor 14 . as a result , compactor 14 could be shut down , either automatically or by an operator , to allow diagnosis and correction of the failure or fault , thereby interrupting the harvesting operation . the methods of the present invention provide alternatives to allow continued operation of the compactor under several sensor fault or failure conditions . referring also to fig4 , a high level flow diagram 76 is shown , including preferred steps for operation of compactor controller 66 in a fault tolerant mode , wherein one or both compactor position sensors 62 a and 62 b have failed or are faulty , and / or a conductive path in connection therewith is faulty . in this mode , controller 66 will determine or set a compactor position value , that is , the position of compactor 14 in relation to some reference , and a compactor tilt value , if any , for use , for instance , in determining the location of compactor 14 in chamber 18 , or whether a movement or indexing of compactor 14 is required . at decision block 78 , controller 66 determines if there is a failure or fault with rear compactor position sensor 62 b . this can be determined from outputs from the sensor , or lack thereof . for instance , a value of an output from the signal can be outside of prescribed range of values , or above or below some prescribed threshold . if a fault is determined , controller 66 will set a rear calculated compactor position value equal to a front calculated compactor position value , as denoted at block 80 . if no fault is determined , controller 66 proceeds to determine if there is a failure or fault with front compactor position sensor 62 a , as denoted at decision block 82 . this determination can be made based on the same or different parameters as those used for determining a fault of sensor 62 b . if a fault is determined , controller 66 sets the front calculated compactor position value equal to the rear calculated compactor position value , as denoted at block 84 . if no fault is determined , controller 66 will proceed to set or determine a compactor position value equal to a value which is a function of a sum of the front calculated compactor position value and the rear calculated compactor position value , as denoted at block 86 . for instance , an average of the position values can be calculated . additionally , controller 66 will set a compactor tilt value equal to a value which is a function of a difference between the rear calculated compactor position value and the front calculated compactor position value . as denoted at decision block 88 , if controller 66 determines that a failure or fault condition exists in regard to both sensors 62 a and 62 b , it will set the compactor position value equal to a default value , which is preferably a bottom position for the compactor , and will set the compactor tilt value equal to a default value , which is preferably zero tilt , as denoted at block 90 . if , at decision block 88 , controller 66 determines that there is no failure of both sensors 62 a and 62 b , the compactor position and tilt values set as indicated in block 86 will be used . as a result , in the event of a fault or failure of one or both of compactor position sensors 62 a and 62 b , or of a conductive path connecting sensors 62 a and 62 b to controller 66 , a compactor position value is determined , such that compactor 14 is still able to operate in a reasonably effective manner . for instance , even if compactor 14 is tilted but a sensor failure makes it impossible to determine the existence of the tilt , controller 66 can operate augers 32 in a default mode . as an example , the augers can be operated in one direction for a first operating period , then reversed and operated in the opposite direction for another time period , such that at least some additional distribution of cotton beneath the compactor will occur before the next compacting operation . a default compaction stroking routine can also be used , such that the cotton in the bottom of the chamber can be compacted to at least some extent , to facilitate receipt of additional cotton into the chamber during continued harvesting . thus , although optimal module building may not occur in the event of failure of one or both of the compactor position sensors 62 a and 62 b , a reasonably effective compaction routine is provided . referring also to fig5 , a high level flow diagram is shown , including preferred steps for operation of compactor controller 66 in a fault tolerant mode in the event of a faulty auger pressure signal , or absence thereof . at block 94 , augers 32 are operating to distribute cotton within compactor chamber 18 . controller 66 will determine if an auger pressure signal is present , as denoted at decision block 96 . if yes , controller 66 will determine whether the signal is good , for instance , within a predetermined or correct value range , as denoted at block 98 . if yes , controller 66 will operate the compactor in the normal manner , for instance , responsive to auger pressure signals , as denoted at block 100 . returning to decision blocks 96 and 98 , if controller 66 determines that the auger pressure signal is not present , or is not within the correct range , controller 66 will operate in a fault tolerance mode , which can involve , for instance , operation of augers 32 for a predetermined auger operation time period . controller 66 will time the operation , and determine when the auger operation time is expired , as denoted at decision block 102 , then will proceed to lower the compactor to perform one or more compacting strokes , as denoted at block 104 . during normal operation , controller 66 will raise the compactor if the compactor pressure exceeds a predetermined threshold . in the event of fault or failure of the compactor pressure sensor 68 , for instance , if the signal from compactor pressure sensor 68 is not within a correct range , or is absent , controller 66 can be programmed to operate in a mode tolerant of this fault , for instance , to raise compactor 14 when a prescribed compactor lowering time exceeds a predetermined threshold . still further , there are some fault conditions wherein continued operation is not desirable until the fault is corrected . for instance , in this regard , controller 66 can be programmed to check the operability or status of critical operator switches during key up or start up , such as , but not limited to , a door open operator switch , a door close operator switch , a fire unload operator switch , and an unload chain operator switch ( not shown ), and if one or more of the switches is faulty , to prevent execution of the operation commanded by the switch , and output or display a message or error signal to the operator , to turn off the switch and / or check the wiring harness , connections , and / or to take other troubleshooting steps . it will be understood that changes in the details , materials , steps , and arrangements of parts which have been described and illustrated to explain the nature of the invention will occur to and may be made by those skilled in the art upon a reading of this disclosure within the principles and scope of the invention . the foregoing description illustrates several preferred embodiments of methods of the invention ; however , concepts , as based upon the description , may be employed in other embodiments without departing from the scope of the invention . accordingly , the following claims are intended to protect the invention broadly as well as in the specific form shown .

a fault tolerant operating method for a cotton compactor of a cotton module builder or packager of a cotton harvesting machine , which serves as an alternative to shutting down the compacting process or erratic operation thereof , in the event of indication of a fault or failure condition involving one or more sensors associated with the compactor , or a conductive path in connection with a sensor , all generally identified as a fault or failure condition .

the disclosure is based on the collection of new data , and a new use of data presently collected within the ensite system as sold by endocardial solutions ( esi ) of st paul minn . in this specification reference is made to patents held by esi , each patent is incorporated by reference herein . the use of the trade marked term ensite is intended to refer to commercially available structures . it has been widely known that one may pace the heart through an ep catheter or through a separate pacing catheter to explore the electrical behavior of the heart during a diagnostic or ablation procedure . more recently it has been determined that pacing in both the left and right ventricle or bi - ventricular pacing is a useful therapy for the treatment of congestive heart failure . by closely coordinating the contraction of both ventricular chambers , an improved cardiac output can be achieved which tends over time to reduce the overt symptoms of congestive heart failure . it is recently , but not widely , recognized that the timing intervals and pacing sites of biventricular pacing must be carefully selected to generate the benefits of biventricular pacing . it is becoming well understood that the precise placement of ventricular pacing leads in the heart is critical to achieving success with biventricular pacing or other pacing therapies directed to patients with chf . it is believed that if the lead system is located in tissue that is refractory , ischemic or scarred , the propagation of activation is delayed and the resulting contraction is disorganized and less effective than normal . the coherence of electrical activation is a non standard but useful way of expressing the requirement that the electrical activation of the heart be propagated over the diseased tissue in a way to result in an effective contraction . from a hemodynamic viewpoint a coherent contraction arises from a homogenous volumetric contraction , in which all portion of the observable heart chamber contract progressively and in “ unison ”. the coherence of electrical activation can be directly observed by the ensite system in the ep data while the homogeneity or hydralic coherence measure is a hemodynamic index computed beat to beat by a modified ensite system . users of the ensite system become skilled at interrupting the propagation of such waveforms and can readily determine the location of infarcted regions in the myocardium based upon their electrical behavior . it is generally wise to avoid attempting to pace these regions of the heart . fig1 shows a commercially available ensite electrophysiology mapping system sold by endocardial solutions of st . paul , minn . although the ensite system in its current commercial embodiments presents electrophysiologic data on a static geometry of the heart , it should be recognized that certain heart information ( ep activation ) is available on a single beat basis this attribute is important in understanding the use of the system in this application . in this system a patient 10 is undergoing a diagnostic procedure through a minimally invasive procedure involving the introduction of an ensite catheter coupled to the breakout box 12 . a conventional electrophysiology catheter 16 is also introduced into the patient while a variety of surface electrodes 11 are used to monitor cardiac activity during the procedure . the breakout box 12 permits the ecg cables and ep system to be coupled to additional hardware , which is not shown in this figure . the patient interface unit 18 couples the ensite catheter to the workstation computer and its related peripherals . 20 . the workstation operates under the control of a software program , which provides a substantial amount of information to the attending physician . in use the physician will see an activation map image similar to that shown in fig3 on the monitor 23 . the computed index 51 will also been shown to the physician as indicated by index value “ 0 . 93 ” seen on the monitor 23 . in general , the physician is able to visualize the intracardiac cavity 32 containing the ensite catheter 14 as seen in fig3 on a color monitor 23 . color is used to reduce the clutter in the image . expressed or displayed on this wire frame geometry image 50 are activation maps and other electrophysiology information derived from the ensite catheter in conjunction with the ep catheter . in this particular instance , the patient is also provided with one or more pacing catheters 24 which are coupled to a temporary pacer 26 through the breakout box 12 . the temporary pacer 26 allows the physician to make measurements while varying the a - v delay and the v — v delay time . pacing rate may be varied to ensure capture . turning to fig2 the heart 30 is shown schematically with a right ventricle 32 containing the ensite catheter 14 and a conventional ep catheter 16 as well as the pacemaker lead 24 . in brief , software running on the workstation 20 in fig2 can create an electrophysiological map of the heart during a single heartbeat as follows . in operation current sourced from a pair of electrodes ( electrode 40 and 42 ) and injected into the heart chamber 32 , chamber . a roving catheter , shown as ep catheter 16 , is located on the endocardial surface 31 toward the exterior of the heart this catheter may be moved widely and may be placed on the interior heart surface along the septum is shown by reference numeral 33 . the injected current is detected through the electrode 44 on the ep catheter 16 . this location is determined and as the catheter is moved about the chamber , complete chamber geometry can be built up by noting the sequential positions of the electrode 44 . incorporated references describe this process in more detail but for purposes of this disclosure a convex hull modeling technique is used to build a statically displayed interior geometry of the heart chamber by selecting certain locations developed from the electrode motion . the convex hull model of the interior chamber of the heart can be smoothed and a representative wire grid displayed to the physician . such a wire grid is shown in fig3 as element 50 . the ensite catheter also carries an array of passive electrode sites typified by electrode site 46 . these electrodes are arrayed around the geometric access of the ensite balloon 47 . at any given instant some of these electrode sites are pointed toward the exterior surface wall 31 and the septal wall 33 . by computing the inverse solution , the electrophysiologic potentials passing along these surfaces can be measured within one beat . reference may be had to u . s . pat . nos . 5 , 297 , 549 ; 5 , 311 , 866 ; 6 , 240 , 307 and 5 , 553 , 611 for further discussion of the inverse solution and the creation of the electrophysiologic map . each of these references is incorporated in its entirety in the present application . in the commercially available ensite system the depolarization wavefront is displayed on a representative geometric surface such as the grid surface 50 of fig3 . the workstation 20 animates this electrophysiology data and the propagation of the electrical way front along the interior surfaces of the heart can be monitored . wavefronts 80 82 and 83 are sequence movements of the stimulus from pacing site 84 seen in fig3 . fig4 shows an equivalent circuit implementation to facilitate a description of conduction volumetry measurements made from an ensite catheter . returning to the geometry of the array on the ensite catheter 14 the interior of the balloon 47 is non - conductive which provides a limited field of view for each of the electrode sites on the surface of the balloon . in essence each electrode responds only to electrical activity bounded by the heart wall , which is directly opposite the electrode site . for example , an electrode such as electrode 46 sees electrical activity and conductance data bounded by the wall 31 and is blind to electrical activity on wall 33 . in a similar fashion , an electrode such as electrode 50 sees only electrical activity occurring on wall surface 33 . by monitoring the voltages on the array electrodes during the pulse , or more particularly measuring the resistance between adjacent columnar pairs of electrodes as indicated by exemplary difference amplifier 86 it is possible to compute the volume of a partial slice 88 of the chamber volume best seen in fig5 . it is important to note that the volume measurement is segmented into several local volumes typified by volume 88 . fig5 shows a slice of chamber volume computed by measuring the difference in resistance between electrodes adjacent along the axis 21 . this view shows that the volume segments are non - overlapping and extend along the axis 21 . the conductance term r is the resistance measured at electrodes in the passive array . this value is directly available to the software in the program , and rho is the conductance of the blood in ohms - centimeters . d represents the distance between adjacent electrode sites in the passive array along the axis 21 . this value is known from the geometry of the ensite catheter . the preferred conduction volumetry algorithms can be computed very fast and the volume changes throughout a single beat of the heart may be tracked . the measurement of chamber volume is most accurate at the mid volume level indicated in fig4 at reference 90 . it is preferred but not required to sum or stack the independent volume measurements to create “ columnar values ” centered on the axis 21 . this is achieved by adding volumes 92 through 96 to create a column volume 90 located near the septum . a similar process is repeated to create a column volume near the wall 31 as shown as a slice 88 in fig5 as well as elsewhere around the chamber . it is believed that the most effective heartbeat will involve the simultaneous and progressive activation of all of the muscle tissue , which should result in a self similar reduction in the measured volume among all of the volume segments measured . fig6 is a display of four representative volume segments of the heart chamber displayed as a function of time . it is expected that eight volumes will be used most effectively . segment 88 may correspond to the antero - lateral volume while the other traces represent other volumes such as the septal ; antero - septal ; anterior ; antero - lateral ; lateral or other volumes defined around axis 21 . the preferred way to compare the self - similarity of the volume waveforms is to cross correlate them statistically . by cross correlation of the values of the segment volumes over time one can compute a number that represents the similarity relationship of the various waveforms to each other . that is if the all the volumes contract identically then they should share the same waveform morphology and be completely self similar . in this instance the index value is unity . real measurements taken have shown that a chf patient in normal sinus rhythm has an index value of about 0 . 8 . and that by manipulating the a - v delay time , location of stimulus and v — v delay interval this index can by increased to about 0 . 9 this is a very significant improvement in the heart contraction . in fig6 a computed value of 0 . 93 is delayed showing improved contraction behavior based on the selected pacing parameters . it is important to note that it is not intended to make a display like fig6 available to the physician because it is difficult to “ compute ” self - similarity qualitatively . the figure is designed to show how the performance index is calculated . index in operation the physician will have the index saved for each pacing location and set of pacing variables . the physician will look for an improved contraction that is reflected by a high index value and a “ normal ” activation sequence . for example a relatively invariant collection of volumes on one side of the heart or the other is some indication that wall is not contracting vigorously and that a better pacing site should be selected . this coherence of contraction index can be displayed as a simple number of percent of a total ( unity ). it is expected that simple figures of merit will be displayed for the physician to allow him to optimize the location of the pacing lead . it is expected that a measure of hemodynamic performance based upon conduction volumetry will be given independently of a coherence of contraction index . it must be recognized that such measures are largely arbitrary and they may be combined in a variety of ways to improve the relationship between the hemodynamic performance index and the clinical outcome for the patient based upon pacing site .

conduction volumetry is used to determine the hemo - dynamic performance of the heart under various pacing protocols to optimize cardiac output as a function of the pacing protocol .

it is to be understood that the figures and descriptions of the disclosed embodiments have been simplified to illustrate elements that are relevant for a clear understanding of the disclosed embodiments , while eliminating , for purposes of clarity , other elements . those of ordinary skill in the art will recognize , however , that these and other elements may be desirable . however , because such elements are well known in the art , and because they do not facilitate a better understanding of the disclosed embodiments , a discussion of such elements is not provided herein . certain embodiments will now be described to provide an overall understanding of the principles of the structure , function , manufacture , and use of the devices and methods disclosed herein . one or more examples of these embodiments are illustrated in the accompanying drawings . those of ordinary skill in the art will understand that the devices and methods specifically described herein and illustrated in the accompanying drawings are non - limiting embodiments and that the scope of these embodiments is defined solely by the claims . the features illustrated or described in connection with one embodiment may be combined with the features of other embodiments . further , where an ordering of steps in a process is indicated , such ordering may be rearranged or the steps may be carried out contemporaneously as desired unless illogical or the listed order is explicitly required . such modifications and variations are intended to be included within the scope of the appended claims . also , in the following description , it is to be understood that terms such as “ forward ,” “ rearward ,” “ front ,” “ back ,” “ right ,” “ left ,” “ over ,” “ under ,” “ top ,” “ bottom ,” “ upwardly ,” “ downwardly ,” “ proximally ,” “ distally ,” and the like are words of convenience and are not to be construed as limiting terms . the description below is for the purpose of describing various embodiments and is not intended to limit the appended claims . turning to the drawings , wherein like numerals denote like components throughout the several views , in fig1 - 2 , a surgical stapling and severing instrument 10 that is capable of practicing the unique benefits of at least one embodiment , including both fluid actuation ( e . g ., opening , closing / clamping ) of an upper jaw ( anvil ) 12 of an end effector 14 as well as dispensing a medical substance onto tissue as severed . fluid actuation of the end effector 14 provides a range of design options that avoid some design limitations of traditional mechanical linkages . for example , instances of binding or component failure may be avoided . further , dispensing liquids onto severed tissue allows for a range of advantageous therapeutic treatments to be applied , such as the application of anesthetics , adhesives , cauterizing substances , antibiotics , coagulant , etc . with particular reference to fig2 , the surgical stapling and severing instrument 10 includes an implement portion 16 formed by an elongate shaft 18 and the end effector 14 , depicted as a stapling assembly 20 . the surgical stapling and severing instrument 10 also includes a handle 22 ( fig1 ) attached proximally to the shaft 18 . the handle 22 remains external to the patient as the implement portion 16 is inserted through a surgical opening , or especially a cannula of a trocar that forms a pneumoperitoneum for performing a minimally invasive surgical procedure . left and right fluid bladders ( lift bags ) 24 , 26 are supported within an aft portion 28 of a staple channel 30 . the anvil 12 includes a pair of inwardly directed lateral pivot pins 32 , 34 that pivotally engage outwardly open lateral pivot recesses 36 , 38 formed in the staple channel 30 distal to the aft portion 28 . the anvil 12 includes a proximally directed lever tray 40 that projects into the aft portion 28 of the staple channel 30 overtop and in contact with the fluid bladders ( lift bags ) 24 , 26 such that filling the fluid bladders 24 , 26 causes a distal clamping section 41 of the anvil 12 to pivot like a teeter - totter toward a staple cartridge 42 held in a distal portion 44 of the staple channel 30 . evacuation and collapse of the fluid bladders 24 , 26 , or some other resilient feature of the end effector 14 , causes the anvil 12 to open . left and right fluid conduits 46 , 48 communicate respectively with the left and right fluid bladders 24 , 26 to bi - directionally transfer fluid for actuation . it should be appreciated that applications consistent with the present embodiment may include a mechanical actuation in the handle 22 ( e . g ., closure trigger ) ( not shown ) wherein the user depresses a control that causes closure and clamping of the end effector 12 . it will be appreciated that the terms “ proximal ” and “ distal ” are used herein with reference to a clinician gripping a handle of an instrument . thus , the staple applying assembly 20 is distal with respect to the more proximal handle 22 . it will be further appreciated that , for convenience and clarity , spatial terms such as “ vertical ” and “ horizontal ” are used herein with respect to the drawings . however , surgical instruments are used in many orientations and positions , and these terms are not intended to be limiting and absolute . with particular reference to fig2 , the elongate shaft 18 includes a frame 50 whose proximal end is rotatably engaged to the handle 22 ( fig1 ) such that a rotation knob 52 rotates the frame 50 along with the end effector 14 . a distal end of the frame 50 has lateral recesses 54 that engage a proximal lip 56 of the staple channel 30 . the frame 50 includes a laterally centered , bottom firing slot 58 that passes longitudinally through the frame 50 for receiving a two - piece firing bar 60 comprised of a firing bar 62 with a distally attached e - beam 64 , the latter translating within the staple applying assembly 20 to sever and staple tissue . a distal portion of the frame 50 includes an upper cavity 66 whose distal and proximal ends communicate through distal and proximal apertures 68 , 70 , defining there between a cross bar 72 over which a distally projecting clip 74 of a clip spring 76 engages with a lower spring arm 78 , distally and downwardly projecting through the upper cavity 66 to bias the firing bar 62 downwardly into engagement with the staple channel 30 , especially when the lower spring arm 78 encounters a raised portion 80 on the firing bar 62 . medical substance dispensing is integrated into the elongate shaft 18 by including a laterally offset cylindrical cavity 90 formed in the frame 50 that communicates along its longitudinal length to the outside via a rectangular aperture 92 that is slightly shorter than an electroactive polymer ( eap ) syringe 100 that is inserted through the aperture 92 into the cylindrical cavity 90 . a proximal portion of the cylindrical cavity 90 contains a longitudinally aligned compression spring 102 that urges a distal dispensing cone 104 of the eap syringe 100 distally into sealing contact with the frame 50 and allows translation for insertion and removal of the eap syringe 100 . an electrical conductor 106 passes through the frame 50 and is attached to the compression spring 102 , which is also formed of an electrically conductive metal . an aft portion of the eap syringe 100 is conductive and contacts the spring 102 to form a cathode to an eap actuator 110 held in a proximal portion of the eap syringe 100 . it will be appreciated that another conductor , perhaps traveling with the conductor 106 , also electrically communicates to the eap actuator 110 to serve as the anode . when activated , the eap actuator 110 longitudinally expands , serving as a plunger to dispel a medical substance 112 in a distal portion of the eap syringe 100 through the distal dispensing cone 104 . insofar as the eap actuator 110 laterally contracts to compensate for its longitudinal expansion , a plunger seal 114 maintains a transverse seal within the eap syringe 100 . an vent ( not shown ), such as around conductor 106 allows air to refill the eap syringe 100 behind the plunger seal 114 as the medical substance 112 is dispensed . the vent may rely upon the surface tension of the medical substance 112 to avoid leaking or be a one - way valve . as described below , the medical substance 112 is conducted by the frame 50 to a lateral fluid groove 120 that is formed in the firing bar 62 and the e - beam 64 to direct the medical substance to a cutting surface 122 of the e - beam 64 . the frame slot 58 is sized to seal the lateral fluid groove 120 . the portion of the lateral fluid groove 120 that is positioned under the spring clip 76 is sealed by a firing bar guide 124 . in the illustrative version , an outer sheath 130 encompasses the frame 50 and proximally projecting lever tray 40 of the anvil 12 . a top distal opening 131 allows closing of the anvil 12 . an outer rectangular aperture 132 of the outer sheath 130 is sized and longitudinally positioned to correspond to the rectangular aperture 92 formed in frame 50 . in some applications , the outer sheath 130 may be rotated to selectively align the rectangular aperture 92 with the outer rectangular aperture 132 for insertion or removal of the eap syringe 100 . it should be appreciated that in some applications that the eap syringe 100 may be integrally assembled into an elongate shaft that does not allow for selecting a desired medical substance . for instance , a disposable implement portion with an integral staple cartridge and medical dispensing reservoir may be selected by the clinician as a unit . it is believed that allowing insertion at the time of use , though , has certain advantages including clinical flexibility in selecting a medical substance ( e . g ., anesthetics , adhesives , antibiotics , cauterizing compound , etc .) and extending the shelf life / simplifying storage and packaging of the implement portion 16 . in the illustrative version , an elongate stack of many disk - shaped eap layers are aligned longitudinally and configured to expand along this longitudinal axis . electroactive polymers ( eaps ) are a set of conductive doped polymers that change shape when electrical voltage is applied . in essence , the conductive polymer is paired to some form of ionic fluid or gel and electrodes . flow of the ions from the fluid / gel into or out of the conductive polymer is induced by the voltage potential applied and this flow induces the shape change of the polymer . the voltage potential ranges from iv to 4 kv , depending on the polymer and ionic fluid used . some of the eaps contract when voltage is applied and some expand . the eaps may be paired to mechanical means such as springs or flexible plates to change the effect that is caused when the voltage is applied . there are two basic types of eaps and multiple configurations of each type . the two basic types are a fiber bundle and a laminate version . the fiber bundle consists of fibers around 30 - 50 microns . these fibers may be woven into a bundle much like textiles and are often called eap yarn because of this . this type of eap contracts when voltage is applied . the electrodes are usually made up of a central wire core and a conductive outer sheath that also serves to contain the ionic fluid that surrounds the fiber bundles . an example of a commercially available fiber eap material , manufactured by santa fe science and technology and sold as panion ™ fiber , is described in u . s . pat . no . 6 , 667 , 825 , which is hereby incorporated by reference in its entirety . the other type is a laminate structure , which consists of a layer of eap polymer , a layer of ionic gel and two flexible plates that are attached to either side of the laminate . when a voltage is applied , the square laminate plate expands in one direction and contracts in the perpendicular direction . an example of a commercially available laminate ( plate ) eap material is from artificial muscle inc , a division of sri laboratories . plate eap material is manufactured by eamex of japan and is referred to as thin film eap . it should be noted that eaps do not change volume when energized ; they merely expand or contract in one direction while doing the opposite in the transverse direction . the laminate version may be used in its basic form by containing one side against a rigid structure and using the other much like a piston . the laminate version may also be adhered to either side of a flexible plate . when one side of the flexible plate eap is energized , it expands flexing the plate in the opposite direction . this allows the plate to be flexed in either direction , depending on which side is energized . an eap actuator usually consists of numerous layers or fibers bundled together to work in cooperation . the mechanical configuration of the eap determines the eap actuator and its capabilities for motion . the eap may be formed into long stands and wrapped around a single central electrode . a flexible exterior outer sleeve will form the other electrode for the actuator as well as contain the ionic fluid necessary for the function of the device . in this configuration when the electrical field is applied to the electrodes , the strands of eap shorten . this configuration of eap actuator is called a fiber eap actuator . likewise , the laminate configuration may be placed in numerous layers on either side of a flexible plate or merely in layers on itself to increase its capabilities . typical fiber structures have an effective strain of 2 - 4 % where the typical laminate version achieves 20 - 30 %, utilizing much higher voltages . for instance , a laminate eap composite may be formed from a positive plate electrode layer attached to an eap layer , which in turn is attached to an ionic cell layer , which in turn is attached to a negative plate electrode layer . a plurality of laminate eap composites may be affixed in a stack by adhesive layers there between to form an eap plate actuator . it should be appreciated that opposing eap actuators may be formed that can selectively bend in either direction . a contracting eap fiber actuator may include a longitudinal platinum cathode wire that passes through an insulative polymer proximal end cap through an elongate cylindrical cavity formed within a plastic cylinder wall that is conductively doped to serve as a positive anode . a distal end of the platinum cathode wire is embedded into an insulative polymer distal end cap . a plurality of contracting polymer fibers are arranged parallel with and surrounding the cathode wire and have their ends embedded into respective end caps . the plastic cylinder wall is peripherally attached around respective end caps to enclose the cylindrical cavity to seal in ionic fluid or gel that fills the space between contracting polymer fibers and cathode wire . when a voltage is applied across the plastic cylinder wall ( anode ) and cathode wire , ionic fluid enters the contracting polymer fibers , causing their outer diameter to swell with a corresponding contraction in length , thereby drawing the end caps toward one another . returning to fig1 , the handle 22 controls closure of the anvil 12 , firing of the two - piece firing bar 60 ( fig2 ), and dispensing of the medical substance . in an illustrative version , a pistol grip 140 may be grasped and a thumb button 142 depressed as desired to control closure of the anvil 12 . the thumb button 142 provides a proportional electrical signal to an eap dispensing actuator not shown ) similar to the eap syringe 100 to transfer fluid through the conduits 46 , 48 to the fluid bladders 24 , 26 to close the anvil 12 ( fig2 ). when the thumb button 142 is fully depressed , a mechanical toggle lock ( not shown ) engages to hold the thumb button 142 down until a full depression releases the toggle lock for releasing the thumb button 142 . thus , when the thumb button 142 is held down , the surgeon has a visual indication that the end effector 14 is closed and clamped , which may be maintained in this position by continued activation of an eap dispensing actuator or by a locking feature . for instance , control circuitry may sense movement of the thumb button 142 , causing a normally closed eap shutoff valve ( not shown ) to open that communicates between the eap dispensing actuator and the conduits 46 , 48 . once movement ceases , the eap shutoff valve is allowed to close again , maintaining the anvil 12 position . in addition , a manual release could be incorporated to defeat such a lockout to open the anvil 12 . as an alternative , a closure trigger ( not shown ) or other actuator may be included that bi - directionally transfers fluid to the fluid bladders 24 , 26 as described in commonly owned u . s . patent application ser . no . 11 / 061 , 908 entitled “ surgical instrument incorporating a fluid transfer controlled articulation mechanism ” to kenneth wales and chad boudreaux , filed on feb . 18 , 2005 , now u . s . pat . no . 7 , 559 , 450 , the disclosure of which is hereby incorporated by reference in its entirety . a number of such fluid actuators for articulation of a pivoting shaft are described that may be adapted for closing the anvil 12 . to take full advantage of the differential fluid transfer described for several of these versions , it should be appreciated that an opposing lift bag ( not shown ) may be placed above the lever tray 40 of the anvil 12 to assert an opening force as the left and right fluid bladders ( lift bags ) 24 , 26 collapse . with particular reference to fig3 , the handle 22 includes a firing trigger 150 ( fig1 ) that is drawn proximally toward the pistol grip 140 to cause a firing rod 152 to move distally in a proximal portion 154 of the elongate shaft 18 . a distal bracket 156 of the firing rod 152 engages an upward proximal hook 158 of the firing bar 62 . a dynamic seal 160 within the frame 50 seals to the firing rod 152 so that the implement portion 16 is pneumatically sealed when inserted into an insufflated abdomen . an anti - backup mechanism 170 of the firing rod 152 may be advantageously included for a handle 22 that includes a multiple stroke firing trigger 150 and a retraction biased firing mechanism coupled to the firing rod 152 ( not shown ). in particular , an anti - backup locking plate 172 has the firing rod 152 pass through a closely fitting through hole ( not shown ) that binds when a retracting firing rod 152 tips the lock plate 172 backward as shown with the bottom of the locking plate 172 held in position within the frame 50 . an anti - backup cam sleeve 174 is positioned distal to the anti - backup locking plate 172 and urged into contact by a more distal compression spring 176 through which the firing rod 152 passes and that is compressed within the frame 50 . it should be appreciated that mechanisms in the handle 22 may manually release the anti - backup mechanism 170 for retraction of the firing rod 152 . in fig4 - 5 , the end effector 14 , which in the illustrative version is a staple applying assembly 20 , is opened by having fluid bladder 24 deflated , drawing down lever tray 40 of the anvil 12 , which pivots about pin 32 raising distal clamping section 41 thereby allowing positioning body tissue 180 between the anvil 12 and staple cartridge 42 . the e - beam 64 has an upper pin 182 that resides within an anvil pocket 184 allowing repeated opening and closing of the anvil 12 . an anvil slot 186 formed along the length of the anvil 12 receives the upper pin 182 when the anvil 12 is closed and the two piece firing bar 60 is distally advanced . a middle pin 188 slides within the staple cartridge 42 above the staple channel 30 in opposition to a bottom pin or foot 190 that slides along a bottom surface of the staple channel 30 . in fig6 - 7 , the staple applying assembly 20 has been closed by expanding the fluid bladder ( lift bag ) 24 , raising the lever tray 40 of the anvil 12 until flush with the outer sheath 130 , with a proximal upwardly bent tip 192 of the lever tray 40 allowed to enter the top distal opening 131 . this bent tip 192 in combination with the opening 131 , advantageously allows greater radial travel for the anvil 12 as well as presenting an abutting surface rather than a piercing tip to the underlying fluid bladder 24 . when the anvil 12 is closed , the upper pin 182 is aligned with the anvil slot 186 for firing and the tissue 180 is flattened to a thickness appropriate for severing and stapling . in fig7 - 8 , the e - beam 64 is cut away to show its bottom foot 190 riding along a downwardly open laterally widened recess 200 that communicates with a narrow longitudinal slot 202 through which a vertical portion 204 of the e - beam 64 passes . a proximal aperture 206 to the narrow longitudinal slot 202 allows an assembly entrance for the lower foot 190 . a bottom bump 208 is positioned on the firing bar 62 to drop into the proximal aperture 206 during an initial portion of firing travel under the urging of the clip spring 76 ( fig6 ) against the raised portion 80 of the firing bar 62 for proper engagement and for possible interaction with an end effector firing lockout mechanism ( not shown ). also , this position allows for the end effector 14 to be pinched shut to facilitate insertion through a surgical entry point such as a cannula of a trocar ( not shown ). with reference to fig8 - 10 , the firing bar guide 124 laterally contacts a portion of the firing bar 62 to close the corresponding portion of the lateral fluid groove 120 . in fig1 , the eap syringe 100 in the cylindrical cavity 90 has its distal dispensing cone 104 communicating with a radial fluid passage 220 formed in the frame 50 that communicates in turn with the lateral fluid groove 120 . in fig1 , before installation in the surgical stapling and severing instrument 10 , the eap syringe 100 may be advantageously sealed with a disposable cap 230 . in fig1 - 14 , the eap syringe 100 is shown without the disposable cap 230 and urged by spring 102 distally to engage the distal dispensing cone 104 into communication with the radial fluid passage 220 . it should be appreciated that one or more sensor in the surgical stapling and severing instrument 10 may sense a firing condition ( e . g ., movement of firing bar or mechanism coupled to the firing bar , position of the firing trigger , a separate user control to dispense , etc .) and activate dispensing control circuitry to effect dispensing . in fig1 - 18 , an alternate two - piece firing bar 300 is formed from longitudinally laminated left half and right half firing bar portions 302 , 304 that form a firing bar 305 and attached to an e - beam 309 . thereby , fluid transfer down the firing bar 300 may be further constrained . in particular , a left side fluid groove 310 in the left half firing bar portion 302 transitions distally to a pair of aligned internal fluid grooves 312 , 314 respectively in the left and right half firing bar portions 302 , 304 , defining an internal fluid passage 316 . since the e - beam 309 is laterally thicker and of short longitudinal length , a drilled fluid passage 320 is formed therein between a cutting surface 322 and an aft edge aligned to communicate with the internal fluid passage 316 . while the present embodiment has been illustrated by description of several embodiments and while the illustrative embodiments have been described in considerable detail , it is not the intention of the applicant to restrict or in any way limit the scope of the appended claims to such detail . additional advantages and modifications may readily appear to those skilled in the art . for example , while a non - articulating shaft is described herein for clarity , it should be appreciated that medical substance dispensing may be incorporated into an articulating shaft . in addition , fluid conduits may be incorporated that pass through an articulation joint of a shaft to fluid bladder actuators that close an end effector . as another example , while both medical substance dispensing and fluid actuated anvil closing are illustrated herein , applications consistent with aspects of various embodiments may include either of these features . further , for applications in which an adhesive and / or cauterizing medical substance is dispensed , it should be appreciated that features such as staples may be omitted . as another example , while a staple applying assembly 20 is illustrated herein , it should be appreciated that other end effectors ( graspers , cutting devices , etc .) may benefit from either or both of fluid controlled closing and medical substance dispensing . as yet another example , a receptacle for the eap syringe may be formed in the handle rather than in the elongate shaft . as yet an additional example , a symmetric arrangement for a second eap syringe may be formed in the elongate channel so that two medical substances may be simultaneously dispensed during firing . as yet a further example , while a staple applying apparatus provides an illustrative embodiment , it should be appreciated that other endoscopic instruments may benefit from the ability to dispense a liquid at or near a distal end thereof . examples of instruments that may benefit include , but are not limited to , an ablation device , a grasper , a cauterizing tool , an anastomotic ring introduction device , a surgical stapler , a linear stapler , etc . as such , those instruments that do not employ a firing bar that serves herein as a convenient fluid passage to a cutting surface may instead incorporate ducting or fluid conduits to an appropriate location . while an electroactive polymer plunger has various advantages , it should be appreciated that other types of actuated devices may be employed to dispense a medical substance to the end effector . for example , fig1 - 20 illustrate various embodiments of a disposable loading unit 1010 , with fig2 showing an exploded view of the disposable loading unit 1010 . the disposable loading unit 1010 includes a first end 1012 configured for releasable connection to a surgical instrument ( see fig3 ), and a second end 1014 opposite the first end 1012 . the disposable loading unit 1010 comprises a housing assembly 1016 , an agent cartridge 1018 , a knife assembly 1020 , a staple cartridge 1022 , and an anvil assembly 1024 . the disposable loading unit 1010 may be removed and discarded after a single use . the housing assembly 1016 comprises a channel 1026 and a channel cover 1028 connected to the channel 1026 . the channel 1026 and the channel cover 1028 may be fabricated from any suitable material such as , for example , a plastic . the channel 1026 includes a first end 1030 proximate the first end 1012 of the disposable loading unit 1010 and a second end 1032 proximate the second end 1014 of the disposable loading unit 1010 . the channel 1026 comprises a base 1034 , a first wall 1036 , and a second wall 1038 . according to various embodiments , the base 1034 defines an opening 1040 proximate the first end 1030 of the channel 1026 , a first slot 1042 proximate the first end 1030 of the channel 1026 , a second slot 1044 proximate the first end 1030 of the channel 1026 , and a third slot 1046 proximate the second end 1032 of the channel . the first wall 1036 is connected to the base 1034 and extends generally perpendicular therefrom . the second wall 1038 is connected to the base 1034 , extends generally perpendicular therefrom , and is opposite the first wall 1036 . the second wall 1038 may be a mirror - image of the first wall 1036 , and the first and second walls 1036 , 1038 may be fabricated integral with the base 1034 . according to various embodiments , each of the first and second walls 1036 , 1038 define a fourth slot 1048 , a first tab 1050 , a first indent 1052 , a fifth slot 1054 , a second indent 1056 , a sixth slot 1058 , a third indent 1060 , a fourth indent 1062 , a seventh slot 1064 , an eighth slot 1066 , and a first flange 1068 . the channel cover 1028 includes a first end 1070 proximate the first end 1012 of the disposable loading unit 1010 and a second end 1072 opposite the first end 1070 , and may be symmetric along an axis that extends from the first end 1070 of the channel cover 1028 to the second end 1072 of the channel cover 1028 . the channel cover 1028 is configured to engage with the channel 1026 at a plurality of locations . according to various embodiments , the channel cover 1028 defines a pair of coupling pegs 1074 proximate the first end 1070 of the channel cover 1028 that extends from the channel cover 1028 . one of the coupling pegs 1074 passes through the opening 1040 defined by the channel 1026 . the channel cover 1028 also defines a slit 1076 proximate the second end 1072 of the channel cover 1028 . according to various embodiments , the channel cover 1028 defines a first pair of tabs 1078 that pass through and engage with the fourth slots 1048 , a first pair of interior projections that mate with the first indents 1052 , a second pair of tabs 1080 that pass through and engage with the fifth slots 1054 , a second pair of interior projections that mate with the second indents 1056 , and a third pair of interior projections that engage with the sixth slots 1058 . according to other embodiments , the channel 1026 and the channel cover 1028 may be fabricated to include other arrangements of tabs , slots , projections , indents , etc . that may be utilized to connect the channel cover 1028 to the channel 1026 . the agent cartridge 1018 is connected to the housing assembly 1016 and houses at least one medical agent . the medical agent may be any type of medical agent . for example , the medical agent may comprise an anesthetic , an adhesive , an antibiotic , a cauterizing substance , a coagulant , a growth hormone , a hemostatic agent , a sealant , etc ., or any combination thereof . the agent cartridge 1018 includes a first end 1082 proximate the first end 1012 of the disposable loading unit 1010 and a second end 1084 opposite the first end 1082 . the agent cartridge 1018 comprises a body 1086 ( see fig2 ) that may be fabricated from any suitable material ( e . g ., a plastic ) that is compatible with the medical agent . according to various embodiments , the body 1086 comprises a first section 1088 and a second section 1090 . the first section 1088 may define a first spline that extends therefrom , and passes through and engages with the first slot 1042 of the base 1034 of the channel 1026 . as shown in fig2 , the first section 1088 may also define a first projection 1094 and a first dispensing port 1096 proximate the second end 1084 of the agent cartridge 1018 . the first projection 1094 may be of any shape ( e . g ., rectangular , triangular , hemispherical , etc .). the first dispensing port 1096 is positioned between the first projection 1094 and the second end 1084 of the agent cartridge 1018 . the second section 1090 is spaced apart from the first section 1088 and may be a mirror - image thereof . the second section 1090 may define a second spline that extends therefrom , and passes through and engages with the second slot 1044 of the base 1034 of the channel 1026 . as shown in fig2 , the second section 1090 may define a second projection 1100 and a second dispensing port 1102 proximate the second end 1084 of the agent cartridge 1018 . the second projection 1100 may be of any shape ( e . g ., rectangular , triangular , hemispherical , etc .). the second dispensing port 1102 is positioned between the second projection 1100 and the second end 1084 of the agent cartridge 1018 . according to other embodiments , the body 1086 may be fabricated to include other arrangements of splines , tabs , fasteners , etc . that may be utilized to connect the agent cartridge 1018 to the housing assembly 1016 . according to various embodiments , the agent cartridge 1018 also comprises a first sealing member 1104 ( see fig2 ) and a second sealing member 1106 ( see fig2 ). the first sealing member 1104 is connected to the first section 1088 and cooperates with the first section 1088 to house a medical agent . similarly , the second sealing member 1106 is connected to the second section 1090 and cooperates with the second section 1090 to house a second medical agent . the first medical agent may be the same or different than the second medical agent . the knife assembly 1020 is connected to the housing assembly 1016 , and includes a first end 1108 proximate the first end 1012 of the disposable loading unit 1010 and a second end 1110 opposite the first end 1108 . the knife assembly 1020 comprises a body 1112 and a cutting surface 1114 . according to various embodiments , the cutting surface 1114 comprises a portion of a knife blade that is connected to the body 1112 proximate the second end 1110 of the knife assembly 1020 . the body 1112 may be fabricated from any suitable material such as , for example , a plastic . according to various embodiments , the body 1112 comprises a first clamping member 1116 proximate the first end 1108 of the knife assembly 1020 , a second clamping member 1118 proximate the first end 1108 of the knife assembly 1020 , and a foot member 1120 proximate the second end 1110 of the knife assembly 1020 . the foot member 1120 passes through the third slot 1046 of the base 1034 of the channel 1026 and is mated with a retainer 1122 that is external to the housing assembly 1016 and serves to slidably connect the body 1112 to the housing assembly 1016 such that the knife assembly 1020 can be selectively advanced along the third slot 1046 toward the second end 1032 of the channel 1026 . the body 1112 of the knife assembly 1020 also comprises a first surface 1124 and a second surface 1126 ( see fig2 ) that is opposite the first surface 1124 . the first surface 1124 of the body 1112 is adjacent the first section 1088 of the agent cartridge 1018 , and the second surface 1126 of the body 1112 is adjacent the second section 1090 of the agent cartridge 1018 . the first surface 1124 of the body 1112 defines a first groove 1128 and the second surface 1126 of the body 1112 defines a second groove 1130 ( see fig2 ). the first groove 1128 is proximate the cutting surface 1114 of the knife assembly 1020 and may extend any distance along the first surface 1124 of the body 1112 toward the first end 1108 of the knife assembly 1020 . the first groove 1128 is adjacent the first dispensing port 1096 and is configured to receive the first projection 1094 of the first section 1088 of the body 1086 . the second groove 1130 is proximate the cutting surface 1114 of the knife assembly 1020 and may extend any distance along the second surface 1126 of the body 1112 toward the first end 1108 of the knife assembly 1020 . the second groove 1130 is adjacent the second dispensing port 1102 and is configured to receive the second projection 1100 of the second section 1090 of the body 1086 . each of the first and second grooves 1128 , 1130 may be of any shape ( e . g ., rectangular , triangular , hemispherical , etc .) suitable for respectively receiving the first projection 1094 and the second projection 1100 . the body 1112 of the knife assembly 1020 may also define an opening 1132 that extends from the first surface 1124 to the second surface 1126 proximate the second end 1110 of the knife assembly 1020 . the staple cartridge 1022 is connected to the housing assembly 1016 . the staple cartridge 1022 includes a first end 1134 and a second end 1136 opposite the first end 1134 . the second end 1136 of the staple cartridge 1022 is proximate the second end 1014 of the disposable loading unit 1010 . the staple cartridge 1022 may be similar to other staple cartridges known in the art . for example , the staple cartridge 1022 may comprise a plurality of surgical fasteners and a plurality of corresponding pushers . according to various embodiments , the staple cartridge 1022 defines a slot 1142 that is aligned with the third slot 1046 of the base 1034 of the channel 1026 and extends from the first end 1134 of the staple cartridge 1022 toward the second end 1136 of the staple cartridge 1022 . the staple cartridge 1022 may also define tabs that extend from the staple cartridge 1022 and pass through and engage with the seventh slots 1064 and the eighth slots 1066 of the channel 1026 , and may further comprise flanges 1146 which frictionally engage the first and second walls 1036 , 1038 of the channel 1026 proximate the second end 1032 of the channel 1026 . according to other embodiments , the staple cartridge 1022 may be fabricated to include other arrangements of tabs , flanges , fasteners , etc . that may be utilized to connect the staple cartridge 1022 to the housing assembly 1016 . the anvil assembly 1024 is connected to the housing assembly 1016 . the anvil assembly 1024 includes a first end 1148 and a second end 1150 opposite the first end 1148 . the second end 1150 of the anvil assembly 1024 is proximate the second end 1014 of the disposable loading unit 1010 . the anvil assembly 1024 may be similar to other anvil assemblies known in the art . for example , the anvil assembly 1024 is moveable between an open position and a closed position , and may comprise an anvil plate 1152 and an anvil body 1154 connected to the anvil plate 1152 . according to various embodiments , the anvil plate 1152 defines a slot 1156 that is aligned with the slot 1142 of the staple cartridge 1022 , and the anvil body 1154 defines a slot 1158 that is aligned with the slot 1156 of the anvil plate 1152 . the anvil plate 1152 may also define a first pair of ears 1160 proximate the first end 1148 of the anvil assembly 1024 and a second pair of ears 1162 positioned between the first pair of ears 1160 and the second end 1150 of the anvil assembly 1024 . one of the ears of the second pair of ears 1162 is engaged with the third indent 1060 defined by the first wall 1036 of the channel 1026 , and the other ear of the second pair of ears 1162 is engaged with the third indent 1060 defined by the second wall 1038 of the channel 1026 . a spring member 1164 or other biasing arrangement may be utilized to urge the anvil assembly 1024 to the open position , and an anvil pin 1166 that passes through the opening 1132 of the knife assembly 1020 may be utilized to urge the anvil assembly 1024 toward the closed position . according to other embodiments , the anvil assembly 1024 may be fabricated to include other fastener arrangements that may be utilized to connect the anvil assembly 1024 to the housing assembly 1016 . the disposable loading unit 1010 may further comprise a first medical agent driver 1168 proximate the first end 1082 of the agent cartridge 1018 and a second medical agent driver 1170 ( see fig2 ) proximate the first end 1082 of the agent cartridge 1018 . according to various embodiments , the first and second medical agent drivers 1168 , 1170 may comprise a portion of a drive block 1172 that is coupled to the knife assembly 1020 at the first end 1108 thereof . for such embodiments , the first medical agent driver 1168 may be configured to slidably fit within the first section 1088 of the body 1086 of the agent cartridge 1018 , and the second medical agent driver 1170 may be configured to slidably fit within the second section 1090 of the body 1086 of the agent cartridge 1018 . according to other embodiments , the first medical agent driver 1168 may comprise an electrically activated polymer that is in contact with the first section 1088 of the body 1086 of the agent cartridge 1018 as shown in fig1 and 14 . similarly , the second medical agent driver 1170 may comprise an electrically activated polymer that is in contact with the second section 1090 of the body 1086 of the agent cartridge 1018 . for such embodiments , each of the first and second medical agent drivers 1168 , 1170 may be electrically connected to a contact 1174 ( see fig3 ) that is proximate the first end 1012 of the disposable loading unit 1010 and is connected to a voltage source . as shown in fig2 , the disposable loading unit 1010 may also comprise a lock member 1176 connected to the drive block 1172 , a retainer 1178 for coupling the lock member 1176 to the drive block 1172 , and a sled 1180 positioned proximate the second end 1110 of the knife assembly 1020 . the drive block 1172 , the lock member 1176 , the retainer 1178 and the sled 1180 may be similar to those known in the art . the disposable loading unit 1010 may further comprise a firing member adapter 1182 connected to the drive block 172 . the firing member adapter 1182 is configured for receiving a firing member that does not comprise a portion of the disposable loading unit 1010 . fig2 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . the first and second clamping members 1116 , 1118 are connected to the drive block 1172 , and the lock member 1176 and the retainer 1178 are also connected to the drive block 1172 . the first medical agent driver 1168 is connected to the drive block 1172 , and the sled 1180 is proximate the second end 1110 of the knife assembly 1020 . the general positions of the shown components relative to the channel 1026 represent the positions of the components prior to the advancement of the firing member ( i . e ., the pre - fire positions ). fig2 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . fig2 is similar to fig2 , and shows that the first medical agent driver 1168 is aligned with the first section 1088 of the body 1086 of the agent cartridge 1018 . the general positions of the shown components relative to the channel 1026 represent the positions of the components prior to the advancement of the firing member ( i . e ., the pre - fire positions ). fig2 illustrates various embodiments of the disposable loading unit 1010 , and shows a cross - section of the disposable loading unit 1010 along line 6 - 6 of fig2 . as shown in fig2 , the first and second dispensing ports 1096 , 1102 may pass through the respective first and second sections 1088 , 1090 at an angle relative to the base 1034 of the channel 1026 . fig2 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . the anvil assembly 1024 is shown in the open position relative to the staple cartridge 1022 in fig2 . the general positions of the shown components relative to the channel 1026 represent the positions of the components prior to the advancement of the firing member ( i . e ., the pre - fire positions ). fig2 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . fig2 is similar to fig2 , but also shows the first section 1088 of the body 1086 of the agent cartridge 1018 . fig2 illustrates various embodiments of the disposable loading unit 1010 , and is an enlarged version of a portion of the disposable loading unit 1010 illustrated in fig2 . fig2 illustrates various embodiments of the disposable loading unit 1010 , and is an enlarged version of a portion of the disposable loading unit 1010 illustrated in fig2 . fig2 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . the general positions of the shown components relative to the channel 1026 represent the positions of the components after the advancement of the firing member ( i . e ., the post - fire positions ). as shown in fig2 , the anvil assembly 1024 is in the closed position , and the post - fire positions of the knife assembly 1020 , the anvil assembly 1024 , the first medical agent driver 1168 , the drive block 1172 , and the lock member 1176 are different than their pre - fire positions relative to the channel 1026 . fig3 illustrates various embodiments of the disposable loading unit 1010 , and is an enlarged version of a portion of the disposable loading unit 1010 illustrated in fig2 . as shown in fig3 , the post - fire position of the first medical agent driver 1168 may be some distance from the first dispensing port 1096 . similarly , the post - fire position of the second medical agent driver 1170 may be some distance from the second dispensing port 1102 . fig3 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . fig3 is similar to fig2 , but shows the first section 1088 of the body 1086 of the agent cartridge 1018 , and also shows the first medical agent driver 1168 embodied as an electrically activated polymer . fig3 also illustrates the conductors 1184 that electrically connect the first medical agent driver 1168 and the contact 1174 . the general positions of the shown components relative to the channel 1026 represent the positions of the components prior to the advancement of the firing member ( i . e ., the pre - fire positions ). fig3 illustrates various embodiments of the disposable loading unit 1010 . for purposes of clarity only , certain portions of the disposable loading unit 1010 are not shown in this figure . fig3 is similar to fig2 , but shows the first medical agent driver 1168 embodied as an electrically activated polymer . fig3 also illustrates the contact 1174 and the conductors 1184 that electrically connect the contact 1174 and the first medical agent driver 1168 . the general positions of the shown components relative to the channel 1026 represent the positions of the components prior to the advancement of the firing member ( i . e ., the pre - fire positions ). fig3 illustrates various embodiments of a surgical instrument 1200 . the surgical instrument 1200 includes a handle assembly 1202 , an elongated body 1204 connected to the handle assembly 1202 , and a disposable loading unit 1010 releasably connected to the elongated body 1204 . the disposable loading unit 1010 may be releasably connected to the elongated body 1204 in any manner . for example , the disposable loading unit 1010 may be releasably connected to the elongated body 1204 via the coupling pegs 1074 described hereinabove . the handle assembly 1202 and the elongated body 1204 may be similar to other handle assemblies and elongated bodies known in the art . for example , the handle assembly 1202 may include means for advancing a firing member that is surrounded by the elongated body 1204 and is utilized to advance the drive block 1172 of the disposable loading unit 1010 . in operation , when the firing member is advanced , the advancement of the firing member causes the drive block 1172 to advance toward the second end 1014 of the disposable loading unit 1010 . as the drive block 1172 advances , the knife assembly 1020 advances toward the second end 1014 of the disposable loading unit 1010 . the advancement of the knife assembly 1020 causes the anvil pin 1166 to cooperate with the anvil body 1154 to urge the anvil assembly 1024 toward the closed position . the advancement of the knife assembly 1020 also causes the sled 1180 to advance toward the second end 1014 of the disposable loading unit 1010 . as the sled 1180 advances , the angled leading edges of the sled 1180 sequentially contact pushers supported within the staple cartridge 1022 , causing the pushers to urge surgical fasteners from the staple cartridge 1022 in a known manner . for embodiments where the first and second medical agent drivers 1168 , 1170 are coupled to the knife assembly 1020 , the advancement of the drive block 1172 advances the first and second medical agent drivers 1168 , 1170 within the first and second sections 1088 , 1090 of the body 1086 toward the second end 1084 of the agent cartridge 1018 . as the first and second medical agent drivers 1168 , 1170 advance , they make contact with the first and second sealing members 1104 , 1106 and urge the first and second medical agents out of the first and second dispensing ports 1096 , 1102 . because the post - fire positions of the first and second medical agent drivers 1168 , 1170 may be some distance from the first and second dispensing ports 1096 , 1102 , some medical agent may still remain housed by the agent cartridge 1018 after the first and second medical agent drivers 1168 , 1170 advance from their pre - fire positions to their post - fire positions . for embodiments where the first and second medical agent drivers 1168 , 1170 are electrically activated polymers , the advancement of the firing member causes an electrical connection to be made with the contact 1174 , causing a voltage to be applied to the first and second medical agent drivers 1168 , 1170 . in response to the applied voltage , the first and second medical agent drivers 1168 , 1170 expand within the first and second sections 1088 , 1090 of the body 1086 of the agent cartridge 1018 and urge the first and second medical agents out of the first and second dispensing ports 1096 , 1102 . with the first projection 1094 and the second projection 1100 serving as stops which restrict the flow of the first and second medical agents along the grooves 1128 , 1130 in the direction toward the first end 1012 of the disposable loading unit 1010 , the medical agents urged out of the first and second dispensing ports 1096 , 1102 advance along the respective grooves 1128 , 1130 toward the cutting surface 1114 of the disposable loading unit 1010 . as the knife assembly 1020 advances along the slot 1142 defined by the staple cartridge 1022 , the staple cartridge 1022 also serves to keep the medical agents in the grooves 1128 , 1130 until the medical agents exit the grooves 1128 , 1130 proximate the cutting surface 1114 . the medical agents are thus effectively delivered to the site of the cutting and stapling . after a single use , the disposable loading unit 1010 is removed from the elongated body 1204 and may be replaced with another disposable loading unit 1010 for another use . this process may be repeated any number of times . therefore , the handle assembly 1202 and the elongated body 1204 connected thereto may be reused any number of times . while several embodiments have been described , it should be apparent , however , that various modifications , alterations and adaptations to those embodiments may occur to persons skilled in the art with the attainment of some or all of the advantages of the various embodiments . for example , according to various embodiments , a single component may be replaced by multiple components , and multiple components may be replaced by a single component , to perform a given function or functions . this application is therefore intended to cover all such modifications , alterations and adaptations without departing from the scope and spirit of the disclosed embodiments as defined by the appended claims . any patent , publication , or other disclosure material , in whole or in part , that is said to be incorporated by reference herein is incorporated herein only to the extent that the incorporated material does not conflict with existing definitions , statements , or other disclosure material set forth in this disclosure . as such , and to the extent necessary , the disclosure as explicitly set forth herein supersedes any conflicting material incorporated herein by reference . any material , or portion thereof , that is said to be incorporated by reference herein , but which conflicts with existing definitions , statements , or other disclosure material set forth herein will only be incorporated to the extent that no conflict arises between that incorporated material and the existing disclosure material .

in various embodiments , a surgical system is disclosed . the surgical system comprises a medicament and a surgical instrument comprising a plurality of staples , a cavity configured to store the medicament , a delivery passage configured to convey the medicament from the cavity to tissue , and a firing system . the firing system comprises a movable member and a firing member , wherein the firing system is configured to cooperatively move the movable member and the firing member to concurrently fire the plurality of staples and deliver at least a portion of the medicament to the tissue owing to advancement of the firing member .

in the description of the embodiments shown in the attached figures the same numerals will be used to refer to like or corresponding parts . referring firstly to fig1 to 4 , fig1 shows a syringe for injections provided with a barrel 2 which slidingly receives a plunger having a cross type section , namely provided with four flat wings 4 , 4 &# 39 ;, 5 , 5 &# 39 ;, placed at 90 ° with respect to one another . according to the present invention the barrel 2 is completely opaque , namely non transparent to light , in order to protect the medicinal substance contained therein against light rays . two measuring scales 6 , 6 &# 39 ;, one made of numerals ascending from 0 to 5 ml and the other made of numerals descending from 5 to 0 ml , are placed in whatever possible way on two aligned wings 4 , 4 &# 39 ; of the plunger 3 , for example through incision , colouring or application of adhesive strips . the rectilinear end 8 of the base of barrel 2 provides a common index for the two scales . the measuring scales shown in fig1 to 3 are readable , when numerals are correctly oriented , by persons using the syringe with the right hand . in these figures it may be clearly seen that the amount of the injected substance can be read directly by the user on the superior scale 6 , while the remaining amount can be read on the inferior scale 6 &# 39 ;; the opposite happens in case the syringe is used for drawing a substance or during the filling of the same with medicinal substance . in case of injection for example , in the positions shown in fig2 and 3 , 1 . 7 and 3 . 6 ml of substance have been respectively injected ( readable on scale 6 &# 39 ;) and in the syringe respectively remain 2 . 3 and 1 . 4 ml of substance . the same happens in case of drawing or filling of the syringe through intake of liquid substance by inverting the function of the two measuring scales 6 , 6 &# 39 ;. in fig4 two further measuring scales 7 , 7 &# 39 ; are foreseen along the sides of wings 4 , 4 &# 39 ; opposite to those shown in fig1 to 3 , which are also provided with inverted numbering and numerals readable by left handed persons . clearly the two further measuring scales 7 , 7 &# 39 ; may be foreseen also on the second pair of flat wings 5 , 5 &# 39 ;. the hypodermic syringe shown in fig1 to 4 is provided with a metallic needle 9 which , in a way per se known , has an enlarged inserted base 10 , also made of metal , enclosed in a plastic bush 11 with inner conical shape , fixed , by means of an interference connection , to a conical tang 12 , foreseen on the tip of barrel 2 . according to the present invention , the sleeve base 10 of the needle is lengthened till covering at least 1 / 4 of the length of the needle , in order to stiffen it and limit its tendency to bend . on the needle 9 a protective cap ( not shown ) is positioned , in a way per se known , and the whole can be packed in a sealed envelope . in fig5 and 6 a 60 ml syringe is shown , which differs from the one in fig1 to 4 both because of dimensions and the fact that plunger 3 has a cylindrical shape . the four measuring scales 6 , 6 &# 39 ;, 7 , 7 &# 39 ;, with numbering from 0 to 60 are therefore positioned on the outer cylindrical surface of plunger 3 . in these fig5 and 6 the needle is not shown so as to indicate the possibility of using the syringe as a simple pump . the syringe according to the invention is preferably constructed of plastic material and is of the throw - away type , although the invention clearly also covers other syringes made of more &# 34 ; valuable &# 34 ; materials , which can be used again , provided that the various parts forming the same are sterilized . while the invention has been illustrated and described herein as embodied in a syringe with blocking capacity , it is not intended to be limited to the details shown , since various modifications and structural changes may be made without departing in any way from the spirit of the present invention .

a syringe comprising a completely opaque barrel and a plunger having at least two measuring scales , wherein one scale includes ascending numeral values and the other includes descending numeral values having a common index made of a rectilinear end of the base of the cylinder .

with reference to the drawings , which are meant to be illustrative and not limiting , the invention will now be described in detail with regard for the best mode and the preferred embodiment . in general , the invention is a cranial distractor comprising two cranial bone plates structured for affixation to opposing cranial segments using mechanical fasteners , adhesive bonding or other suitable means , the bone plates being connected to a linear distraction mechanism defining a longitudinal axis whereby operation of the distraction mechanism increases the distance between the bone plates . each of the bone plates is connected to the distraction mechanism by a ball and socket connector assembly such that each bone plate may tilt up and down , rotate about the axis defined by the ball and socket connector assembly , and swivel side to side . with this structure the cranial plates of the distractor will properly orient relative to whatever convex , planar or concave surface topology is presented by a particular area of the cranium during the distraction procedure . the cranial distractor comprises a distal cranial bone plate member 21 and a proximal cranial bone plate member 22 connected or mounted to a linear distraction mechanism 10 , the distraction mechanism 10 operating in known manner such that the distance between the distal cranial plate 21 and the proximal cranial plate 22 can be gradually increased . the distal and proximal cranial plates 21 and 22 are relatively thin in cross - section , and may be composed of a malleable material to allow the cranial plates 21 and 22 to be shaped to conform to the surface topography of the cranial segments to which they will be affixed . the cranial bone plates 21 and 22 are provided with bone screw receiving apertures 24 that allow the plates 21 and 22 to be affixed to opposing cranial bone segments in known manner using mechanical screw fasteners , either permanent or bioresorbable . the cranial bone plates 21 and 22 are sufficiently dimensioned in length and width so as to be readily attachable to the exterior of the cranial bone segments such that localized stresses are reduced . the cranial bone plates 21 and 22 are connected to the distraction mechanism 10 in a manner that allows for independent movement of each cranial plate 21 and 22 in multiple directions relative to the distraction mechanism 10 . depending abutment or tab members 24 may be provided which extend from the proximal side of the distal cranial plate 21 and from the distal side of the proximal cranial plate 22 . these abutment members 24 abut against the sides or edges of the opposing cranial bone segments during the distraction process to further minimize local stresses . in the embodiment shown in fig1 through 3 , the linear distraction mechanism 10 comprises a distal end 14 , a proximal end 15 and an elongated sleeve member 11 defining a longitudinal axis 99 and provided with a longitudinal slot 16 . a threaded drive rod 12 is received coaxially within the sleeve member 11 in a manner that allows for rotation of the drive rod 12 relative to the stationary sleeve member 11 . a head member 13 attached to the proximal end of the drive rod 12 extends beyond the proximal end 15 of the sleeve member 11 and is used to rotate the drive rod 12 , whether by finger manipulation or with the use of a drive tool . the distal cranial plate 21 is joined to the distal end of the sleeve member 11 with a stationary ball and socket connector assembly 31 , such that the stationary ball and socket assembly 31 does not move axially along the distraction mechanism 10 . the ball and socket connector assembly 31 allows the distal cranial plate 21 to move in multiple directions relative to the axis 99 of the distraction mechanism 10 , as shown by the arrows in the drawings . the distal cranial plate 21 may tilt up and down relative to the axis 99 of the distraction mechanism 10 , swivel left and right relative to the axis 99 of the distraction mechanism 10 , and rotate about the distraction mechanism axis 99 , the axis defined by the ball and socket assembly 31 being coaxial to the distraction mechanism axis 99 , such that the cranial plate 21 can be properly oriented for affixation to the cranial bone segment without creating detrimental stresses on the distraction mechanism 10 or the bone screws or other mechanical fasteners . the proximal cranial plate 22 is joined to the drive rod 12 with a moving ball and socket connector assembly 32 , and the moving ball and socket connector assembly 32 is connected to an internally threaded riding collar member 33 which is coaxially mounted onto the externally threaded drive rod and which extends through the slot 16 of the elongated sleeve member 11 . as the drive rod 12 is rotated , the riding collar member 33 and the moving ball and socket connector assembly 32 moves longitudinally along the drive rod 12 in the axial direction . the moving ball and socket connector assembly defines a lateral or radial axis 98 extending outward to the distraction mechanism axis 99 , and the moving ball and socket connector assembly 32 allows the proximal cranial plate 22 to move in multiple directions relative to the axis 98 , as shown by the arrows in the drawings . the proximal cranial plate 22 may tilt up and down relative to the axis 98 , swivel left and right relative to the axis 98 , and rotate about the axis 98 , such that the cranial plate 22 can be properly oriented for affixation to the other cranial bone segment without creating detrimental stresses on the distraction mechanism 10 or the bone screws or other mechanical fasteners . furthermore , the cranial plates 21 and 22 are independently adjustable relative to each other . with this structure , the cranial plates 21 and 22 are readily mountable to the cranial segments in proper conformation and are free to change orientation as they are gradually separated during the distraction procedure . this freedom accommodates the curvature of the cranium such that undesirable stresses are not imparted to the cranial bone segments and the configuration of the bone regeneration is able to approximate a natural curvature . as shown in the alternative embodiment of fig4 , the affixation of the cranial plates 21 and 22 may be reversed such that the proximal cranial plate 22 is mounted by a fixed ball and socket connector assembly 31 to the proximal end 15 of the distraction mechanism and the distal cranial plate 21 is mounted by a moving ball and socket connector assembly 32 onto the internally threaded collar 33 positioned on the externally threaded drive rod 12 , such that the distal cranial plate 21 moves longitudinally along the axis 99 away from the proximal cranial plate 22 . it is contemplated and understood that the cranial distractor may comprise distraction mechanisms having different operational mechanisms as long as the cranial plates are connected to the distractor assembly using ball and socket connector assemblies . furthermore , while axial and lateral mountings for the ball and socket assemblies are preferred in order to minimize the overall height of the device such that the distractor mechanism remains close to the cranium , it is understood that the ball and socket assemblies could also be mounted beneath the distractor mechanism in an alternative embodiment , and the longitudinal slot could be disposed on the underside of the distractor mechanism . it is to be understood that equivalents and substitutions to certain elements set forth above may be obvious to those ordinary skill in the art , and therefore the true scope and definition of the invention is to be as set forth in the following claims .

a pair of bone plates adapted for attachment to cranial bone segments , the cranial plates being mounted to a distraction mechanism such that the cranial plates can be separated gradually in very small increments . each of the cranial plates is mounted to the distraction mechanism with a ball and socket connector assembly , such that each of the plates can move in multiple directions and orientations relative to the distraction mechanism to better accommodate a convex , planar or concave surface topography presented during the distraction procedure .

referring to fig1 a tourniquet cuff according to the preferred embodiment of the invention is shown . the cuff 5 includes an inflatable member 10 , a means 20 for securing the inflatable member around a body part , and a means 30 for facilitating inflation of the inflatable member 10 . the inflatable member 10 has a length dimension l -- l &# 39 ;, a thickness dimension , which is perpendicular to the plane of the drawing of fig1 and a width dimension w -- w &# 39 ;. according to the invention , the inflatable member 10 has a substantially arcuate shape with a radius r of the arc passing along the width dimension . note that in fig1 r is not the full radius of the arc but is only intended to show the direction of a radius . turning now to a more detailed description of the preferred embodiment of the invention , we refer to fig1 and 2 . in the preferred embodiment , inflatable member 10 includes a flexible backing 12 , 14 and a stiffener 13 . stiffener 13 is positioned between the two pieces 12 and 14 of backing material and the edges of the backing material are sealed , as at 42 , to form an envelope containing stiffener 13 . the preferred embodiment of inflatable member 10 also includes a bladder 15 and a bladder protective layer 17 . bladder 15 is preferably formed of two sheets 16a and 16b which are sealed along their edges . protective layer 17 is placed over bladder 15 and both are sealed to the envelope formed by backing pieces 12 and 14 so that bladder 15 is enclosed in an envelope formed by piece 14 and protective layer 17 . it is noted that the thicknesses of the elements 12 , 13 , 14 , 15 , and 17 making up the inflatable member and the relative distance between these pieces has been exaggerated in fig2 in order to more clearly show the individual pieces and the relationships between the pieces . thus the thickness t -- t &# 39 ; is not to be taken to indicate the true thickness of the uninflated inflatable member as compared to the width w -- w &# 39 ; and length l -- l &# 39 ;. in an actual cuff , the thickness t -- t &# 39 ; would be substantially less as compared to the width and length . turning now to a detailed description of the preferred embodiment of the means 20 for securing the inflatable member 10 to a body part , we return to fig1 . the preferred embodiment of means 20 includes a fabric hook and loop fastener system 21 and 22 and a ribbon 23 which is attached to backing 12 by attachment strip 24 . in the embodiment shown , 21 comprises an adhesive - backed strip of fabric hooks , with the adhesive surface applied to the upper surface ( the surface facing out of the paper ) of backing 12 . adhesive - backed strip 22 of fabric loops is attached to the lower surface of backing material 12 . in the drawing the strip 22 is shown in dotted outline since it is on the reverse side of the strip 12 and would not be seen in the view . ribbon 23 and attachment strip 24 form a secondary means for securing the cuff . ribbon 23 is placed against the back side of backing 12 and strip 24 is placed over the ribbon and sealed to backing 12 to attach the ribbon 23 to backing 12 . again , strip 24 is dotted to indicate that it is on the reverse side of the backing 12 and would not be visible from the front side . turning now to a detailed description of the means 30 for facilitating the inflation of inflatable member 10 , means 30 includes tubing 31 and 32 which extend under the edge of protective layer 17 and through the seal 41 of the two sheets 16a and 16b forming bladder 15 . a corner of bladder 15 is shown in dotted outline in fig1 to indicate the connection of tubes 31 and 32 . the ends 36 and 37 of tubes 31 and 32 within bladder 15 are open . the opposite ends of tubes 31 and 32 are connected to male connectors 33 and 34 for connecting to an inflation and deflation means ( not shown ). in the preferred embodiment shown , one of the tubes 31 and 32 is intended to be connected to a source of pressurized gas , while the other one of the tubes is connected to a deflation valve . in other embodiments , there is only one tube which is used for both inflation and deflation . turning now to a description of materials out of which the preferred embodiment is made , the inner and outer layers 12 and 14 of the backing envelope 11 and the attachment strip 24 are preferably made of a lamination of b . f . goodrich koroseal ™ reinforced with polyester fabric . the material used is a lamination known under the trademark weblon . the preferred fabric weight is 12 oz . per square yard , with a grab tensile strength of 120 × 120 pounds per inch , and a tear strength of 24 × 24 pounds . the polyester preferably has a yarn count of 24 × 24 and a denier of 220 . the total vinyl thickness is preferably 10 mil . stiffener 13 is preferably made of 0 . 030 inch polypropylene sheeting , medical grade . the bladder sheets 16a and 16b and the protective layer 17 are preferably made of 0 . 012 mil . translucent pvc ( poly vinyl chloride ). the hook and loop fastener system 21 and 22 is preferably made of adhesive - backed velcro hook and loop fabric . tubing 31 and 32 is preferably 3 / 16 &# 34 ; o . d .× 1 / 8 &# 34 ; i . d . clear medical grade tubing made of a material such as pvc , silicone rubber , etc . connectors 33 and 34 are preferably white nylon , male , luer - compatible connectors . fig4 shows a diagram of a cuff such as cuff 5 of fig1 when it is wrapped around a human limb 4 , such as an arm or a leg . as shown , when the cuff 5 is wrapped in a circle about the limb 4 , it forms a frustum of a cone . this conical shape of the cuff 5 as applied conforms to the general conical nature of limbs 4 . in the preferred embodiment , the proximal end 5a and the distal end 5b of cuff 5 are indicated in order to ensure proper placement of the cuff 5 on limb 4 . the incorporation of a polyester reinforcement in the cuff layers eliminates the possibility of stretch or growth of the backing envelope 11 during inflation . the elimination of stretching further adds to the ability of the cuff to occlude blood at lower pressure setting . the stiffening layer 13 as well as the polyester reinforcement in the pvc force the &# 34 ; growth &# 34 ; of the bladder during inflation to occur only in the direction of the limb . this further eliminates the &# 34 ; ballooning &# 34 ; or &# 34 ; rubber band &# 34 ; effect which cause concentration of pressures at the center of the cuff - to - limb interface . the stiffener 13 further prevents rolling of the cuff along the limb in the distal direction . this feature also decreases the cuff inflation time . on the other hand , because of the strength and stiffness of backing envelope 11 and stiffener 13 , the bladder 15 and protective layer 17 may be made out of relatively soft material , which prevents tissue damage during inflation . it has been found that the protective layer 17 also helps to eliminate ballooning and protects bladder growth at extremely high pressures . another feature of the invention is that all the materials used in the cuff are water or solution repelling and thus there is very little danger of the cuff harboring and absorbing chemicals and solutions that might cause skin - side burns and irritation . turning now to fig3 a set of cuffs 5 through 9 according to the invention is shown . by measuring the arms and legs of a large number of people , a preferred set of sizes of cuffs according to the invention required to conform to as many limb sizes as possible , was determined . it was discovered that by nesting the various sizes , it was possible to provide a set of cuffs which fit nearly every conceivable limb size . by nesting it is meant that the radius of the arc of a smaller size cuff is smaller than the radius of the arc of a larger size cuff . that is , of the cuffs shown in fig3 the arc of cuff 5 would have the largest radius , while the arc of cuff 9 would have the smallest radius , and intermediate cuffs 6 , 7 , and 8 would have intermediate radii . in fig3 ribbon 23 and tubing 31 and 32 are shown for cuff 5 in order to orient the drawing , but other details of cuff 5 and the details of cuff 6 through 9 are not shown . in the preferred embodiment of the invention the inside radius of one size cuff corresponds to the outside radius of another size cuff . this arrangement has been found to conform well to various limb tapers , however it is not necessary . the principles of the invention may be utilized by making , say , the inside radius of a smaller size cuff smaller than the inside radius of a larger size cuff , or for example , the mean radius of a smaller size cuff smaller than the mean radius of a larger size cuff . the following table indicates the preferred dimensions for the embodiment of the set of cuffs described . __________________________________________________________________________ w - w &# 39 ; t r d - d &# 39 ; l - l &# 39 ; bladder ( cuff (° of ( inside ( adhesive ( overalllength width ) taper ) radius ) area ) length ) __________________________________________________________________________ 8 23 / 8 17 ° 20 . 8 3 . 4 12 . 512 3 22 ° 23 . 8 3 . 4 16 . 018 4 26 ° 27 . 8 4 . 8 23 . 324 4 27 ° 31 . 8 5 . 4 29 . 434 4 32 ° 35 . 8 6 . 0 39 . 2__________________________________________________________________________ all dimensions are given in inches except the degree of taper which is given in degrees . the various sealed joints , such as 41 , 42 , 43 , and the attachment of strip 24 to backing material 12 are made by radio frequency ( rf ) sealing . a novel tourniquet cuff that provides more even pressure distribution to the limb and numerous other features and advantages as compared to prior art cuffs has been described . while the above description of the invention has been referenced to a few particular embodiments , it is evident that , now that the advantages of an arcuate cuff shape and the nesting of a set of cuffs has been disclosed , those skilled in the art can now make numerous uses of , modifications of , and departures from the specific embodiments described herein without departing from the inventive concepts . for example , one or another of the various parts of the preferred cuff , such as the stiffener 13 or the protective layer 17 , may be eliminated . other means may be used for securing to the body , or other means for facilitating inflation may be incorporated . a wide variety of cuff dimensions may be used . consequently , the invention is to be construed as embracing each and every novel feature and novel combination of features within the appended claims .

this invention relates to an improved tourniquet cuff . the cuff is of the type which includes an inflatable member for wrapping about a body part , such that the inflatable member is secured to the body part , and at least one tube is provided to facilitate inflation of the inflatable member , with the inflatable member having a length dimension , and a width dimension . the inflatable member is formed in a substantially arcuate shape , with the radius of the arc passing along the width dimension . in the preferred embodiment , the inflatable member includes a bladder and a polyester reinforced backing , both of which have the arcuate shape . the invention also provides a set of tourniquet cuffs having the arcuate shape and in which the cuffs of the set nest one within the other . in the preferred embodiment , the inside radius of one cuff is equal to the outside radius of the next smaller size cuff .

referring now to fig1 , a front and rear unloading forage transporter 10 may include a forage box 12 providing a generally horizontal floor 14 and left and right upstanding sidewalls 16 and 18 defining a volume therein for receiving forage . the forage transporter 10 may optionally include a top 20 attached to the left and right upstanding sidewall 16 and 18 to partially cover the forage - receiving volume . the forage box 12 may be open at a front face 22 and a rear face 24 . the rear face 24 may be closed by pivoting door 26 hinging about a horizontal axis at the upper edge of the door ( as shown ) or two doors pivoting about vertical axes as hingeably attached to the left and right upstanding sidewalls 16 and 18 respectively ( not shown ). generally the forage box 12 will be attached to a wheel set 27 , for example , to produce a forage wagon ( as shown ) which may be drawn by a draw bar 28 or as part of an integrated rear bed of a truck or the like ( not shown ). positioned at a front and rear edge of the floor 14 and generally parallel to the front and rear edges of the floor 14 are front and rear conveyor driveshafts 30 and 32 which may rotate along generally horizontal axes parallel to the floor 14 and aligned with their width . apron chains 34 connected in a loop between and around the front and rear conveyor driveshafts 30 and 32 present a first extent above the floor 14 and a second extent returning below the floor 14 ( not visible in fig1 ). in one embodiment , the apron chains 34 are provided in pairs , each pair connected by slats 36 extending perpendicularly to the extent of the apron chains 34 which provide upwardly extending projections that may engage with the forage in the forage box 12 . rotation of the front and rear conveyor driveshafts 30 and 32 in one direction will cause the apron chains 34 to move so as to transport forage in one direction , for example , out of the rear of the forage box 12 , and rotation of the front and rear conveyor driveshafts 30 and 32 in the opposite direction will cause the apron chains 34 to move so as to transport forage in the opposite direction , for example , out of the front of the box 12 . the rear conveyor driveshaft 32 may connect to a rear drive mechanism 38 being , in one embodiment , a hydraulic motor as will be described below . this hydraulic motor may receive pressurized hydraulic fluid from an external source through hydraulic lines 40 , for example , connected to a hydraulic pump in a tractor or truck or the like . the front conveyor driveshaft 30 connects to a front drive mechanism 42 , as will be described in more detail below , which receives power from a power takeoff ( pto ) driveshaft 44 extending forward from the front of the transporter 10 along a front - to - rear axis 45 to be received by a corresponding pto output shaft of a tractor or truck ( not shown ). the pto driveshaft 44 is received by an independent outfeed clutch 46 as will also be described below . the front drive mechanism 42 , as well as driving the front conveyor driveshaft 30 , may drive a set of three feed rollers 48 , for example augers , in rotation about a set of vertically displaced horizontal axes at the front face 22 of the feed box 12 . generally drive mechanism 38 may directly rotate the lowest most feed roller 48 through a direct coupling shaft and power may be transmitted to the other to feed rollers 48 through a set of overlapping chain drives 50 . the front drive mechanism 42 may further provide power to a cross conveyor 52 positioned beneath and in front of the feed rollers 48 to receive outflows from the feed rollers 48 and move them in a direction perpendicular to the front - to - rear axis 45 . referring now to fig2 , the outfeed clutch 46 may provide a pair of side - by - side pulley 54 and 56 rotating generally about axes parallel to the front - to - rear axis 45 and roughly aligned with the axis of the pto driveshaft 44 , the latter of which connects directly to pulley 54 . the pulleys 54 and 56 are joined by a v - belt 49 of the reinforced elastomeric material that fits loosely about the pulleys 54 and 56 when the clutch 46 is released and which , when the clutch 46 is engaged , is tightened around the pulleys 54 and 56 by an idler wheel 60 moved against the v - belt 49 by a clutch actuation lever 62 that may be manually engaged by operator . when the outfeed clutch 46 is engaged , a chain drive sprocket 64 attached coaxially with pulley 56 drives a chain 66 which engages a cross feed conveyor drive pulley 68 driving the cross conveyor 52 ( shown in fig1 ) and a auger drive sprocket 70 forming part of the front drive mechanism 42 . referring now to fig3 and 4 , the auger drive sprocket 70 is attached to dual groove v - belt pulley 72 so that the two rotate together yet freely about a horizontal jack shaft 74 having an axis also parallel to the axis 45 . the pulley 72 connects via a dual groove v - belt 77 with a corresponding coplanar dual groove v - belt pulley 76 displaced vertically from pulley 72 and attached to auger driveshaft 78 to rotate auger driveshaft 78 . auger driveshaft connects to right angle drive 80 which communicates with the lowermost feed roller 48 as also shown in fig1 . auger driveshaft 78 also connects with a clutch / variable speed drive c / vsd 79 formed by a master split sheave v - belt pulley 82 having co - rotating and separable opposed frustoconical sheaves 83 . pulley 82 is attached to auger driveshaft 78 to rotate therewith and a servant variable sheave belt pulley 84 as joined by single groove v - belt 86 . as is generally understood in the art , by changing the separation between the sheaves of the master pulley 82 , the effective diameter of the master pulley 82 may be changed controlling an effective “ gear ” ratio between pulleys 82 and 84 . separation of the sheaves of the pulley 82 beyond a certain point provides a clutch action effectively disengaging pulley 82 from 84 . the separation of the sheaves 83 of the master pulley 82 is controlled by a pivoting lever 90 driven by a control shaft 92 . as best shown in fig4 , the control shaft 92 terminates at a crank 93 communicating with an arm 97 to one end of lever 90 , the other end of which attaches to one of the sheaves 83 of pulley 82 . the sheaves of the servant pulley 84 are spring biased by helical compression spring 95 to accommodate the changing effective diameter of pulley 82 by reducing or expanding the effective diameter of servant pulley 84 , passively , against the spring biasing of compression spring 95 . servant pulley 84 is attached to jack shaft 74 to rotate therewith and to drive a chain drive 94 communicating with right angle gear drive 96 , the latter which may communicate rotative power directly to front conveyor driveshaft 30 . it will be generally understood then that power is directly conducted from the pto driveshaft 44 to the feed rollers 48 when the outfeed clutch 46 ( shown in fig2 ) is engaged . that power is only then applied to the front conveyor driveshaft 30 with engagement of civsd 79 . this design allows the operator to control the rate of feed forage into the feed rollers 48 while ensuring that the feed rollers 48 will be up to speed before any forage is directed into them . the v - belts 77 and 86 , as well as transferring power , provide for torsional shock absorption . referring now to fig2 and 5 , the control shaft 92 may include a central shaft 100 surrounded by a coaxial tubular shaft 102 , both extending generally perpendicular to axis 45 across a front of the transporter 10 . the central shaft 100 attaches directly to the crank 93 and extends rightward ( as depicted in fig2 ) to a control operator 104 in the form of an l - shaped lever extending radially from the end of the control shaft 92 . the lever may be attached to a splined collar 106 that may slide along central shaft 100 and rotate therewith as a result of entering gauging splines on the central shaft 100 and splined collar 106 . the collar 106 may further be connected to the outer tubular shaft 102 through a swivel coupling 108 . in this way , axial motion 110 of the control operator 104 is transmitted through the outer tubular shaft 102 without affecting the inner shaft 100 , whereas rotational motion 112 of the control operator 104 is transmitted through the central shaft 100 to the crank 93 without affecting the outer tubular shaft 102 . a guide cowling 113 provides a slot 114 constraining motion of the control operator 104 to be able to move axially only when the control operator 104 is fully raised and to move rotationally only when the control operator 104 is in its full rightmost axial position . as depicted in fig5 , when the control operator 104 is in its full leftmost and full raised position , the crank 93 will be positioned to fully disengage the sheaves 83 of pulley 82 ( shown in fig4 ) thus effectively disconnecting front conveyor driveshaft 30 ( shown in fig1 ) and allowing the front conveyor driveshaft 30 to freewheel . the outer tubular shaft 102 will be in its full leftmost position so that a tab 116 extending from the tubular shaft 102 will be removed from an operator 118 of a hydraulic valve 120 . this allows the hydraulic valve 120 to open allowing hydraulic fluid 122 to pass into hydraulic motor 124 forming the rear drive mechanism 38 and attached to the rear conveyor driveshaft 32 . thus , the front conveyor driveshaft 30 will be disconnected and the rear conveyor driveshaft 32 fully connected to rotate under the influence of received hydraulic fluid . this in turn allows discharge of forage from a rear of the forage box 12 through rearward motion of the upper extent of the apron chains 34 ( shown in fig1 ). referring now to fig6 , control operator 104 may be retracted or moved to rightward in the axial direction as guided by slot 114 to an intermediate position . this retraction does not rotate shaft 100 and thus leaves the front conveyor driveshaft 30 disengaged from power but does retract outer tubular shaft 102 so that tab 116 engages operator 118 stopping hydraulic fluid flow and thus motion of hydraulic motor 124 . the hydraulic motor 124 may include an internal relief valve or a ratchet coupling allowing it to freewheel in a backward direction when hydraulic fluid flow ceases . thus , in this intermediate position , neither the front conveyor driveshaft 30 nor the rear conveyor driveshaft 32 are engaged with driving power and may freewheel . it will be appreciated that an alternative type of hydraulic valve 120 may be used and the tab 116 may be repositioned so that removal of engagement of the tab 116 and operator 118 will cause a closing of the valve 120 and that the selection of a particular type valve 120 is simply a matter of engineering choice . referring to fig7 , the operator 104 , once at the intermediate position , may be pulled downward in rotation by a variable amount to rotate the crank 93 to engage the c / vsd 79 ( shown in fig4 ) and thus to cause rotation of the front conveyor driveshaft 30 starting at low rates of rotation and increasing that rotational rate with further rotational movement of the operator 104 downward in rotational motion 112 . this rotation downward of the operator 104 , by virtue of the swivel coupling 108 , does not cause tab 116 to move away from operator 118 of valve 120 and thus holds hydraulic motor 124 in its off state . it will be appreciated that the slot 114 requires this simultaneous disengagement of the front conveyor driveshaft 30 and rear conveyor driveshaft 32 before engagement of the front conveyor driveshaft 30 may be undertaken . in the above discussion , it will be understood that although chains are described for use in the apron conveyor , that other similar materials may be used including reinforced belts and the like all of which will generally be termed belts . further , while the above description shows a single control lever , it will be appreciated that alternative operator configurations may be used including , for example , two operator elements mechanically interlocked to act as one lever with respect to locking out simultaneous activation of the front and rear drive rollers . in addition , the term “ gear ratio ” should be understood as not requiring gears but refer equally to the variable diameter belt drives and the like . certain terminology is used herein for purposes of reference only , and thus is not intended to be limiting . for example , terms such as “ upper ”, “ lower ”, “ above ”, and “ below ” refer to directions in the drawings to which reference is made . terms such as “ front ”, “ back ”, “ rear ”, “ bottom ” and “ side ”, describe the orientation of portions of the component within a consistent but arbitrary frame of reference which is made clear by reference to the text and the associated drawings describing the component under discussion . such terminology may include the words specifically mentioned above , derivatives thereof , and words of similar import . similarly , the terms “ first ”, “ second ” and other such numerical terms referring to structures do not imply a sequence or order unless clearly indicated by the context . when introducing elements or features of the present disclosure and the exemplary embodiments , the articles “ a ”, “ an ”, “ the ” and “ said ” are intended to mean that there are one or more of such elements or features . the terms “ comprising ”, “ including ” and “ having ” are intended to be inclusive and mean that there may be additional elements or features other than those specifically noted . it is further to be understood that the method steps , processes , and operations described herein are not to be construed as necessarily requiring their performance in the particular order discussed or illustrated , unless specifically identified as an order of performance . it is also to be understood that additional or alternative steps may be employed . it is specifically intended that the present invention not be limited to the embodiments and illustrations contained herein and the claims should be understood to include modified forms of those embodiments including portions of the embodiments and combinations of elements of different embodiments as come within the scope of the following claims . all of the publications described herein , including patents and non - patent publications , are hereby incorporated herein by reference in their entireties .

a forage transporter allowing both front and rear unloading using an apron conveyor passing along the floor of the forage transporter provides a single control lever selectively enabling a forward drive for forward unloading and a rear drive for rear unloading while providing a mechanical lockout preventing simultaneous activation of the front and rear drives .

referring to the drawings comprising fig1 - 18 , wherein like numerals indicate like elements , there are shown in fig1 - 12 various embodiments of the heart massager of the present invention , whereas fig1 - 18 show the anatomical elements of the body related to the method of the present invention . the instant invention is related to massagers and a particular method for their usage that allows each massager to be inserted through a small surgical opening made in the chest wall , so as to allow the massager to directly contact the anterior antero - lateral surface of the human heart . each of the massagers of the present invention is manually operated to accomplish a direct substernal massage of the heart so as to induce alternate cycles of compression and relaxation ; thereby , sustaining blood circulation to facilitate resuscitation of the heart from its cardiac arrest condition . manual compression and relaxation of the massager produce pulsatile blood - flow patterns similar to the normal ventricular filling and contraction of a healthy heart . in various embodiments , features are provided that allow for direct internal cardiac defibrillation , detection of abnormal electrocardiac rhythms , cardiac pacing and / or relocation of the massager during the heart massage , to optimize results obtained through manual compression of the massager . the massager has various embodiments , all with padded - concave members having features particularly suited to accommodate different aspects of the heart massage . in one embodiment , an umbrella arrangement is provided which holds a padded cup member in its collapsed condition until the device is subcutaneously introduced through a surgical incision in the intercostal space , and , then , allows for the radial expansion of the sidewall of the cup member so that it directly contacts the heart . in other embodiments , various means are provided to prevent the massager from being pressed downward with excessive force or to rapidly , each of which might otherwise damage or scar the heart being massaged . still further embodiments are provided that have concave members to contact the heart in a conformal manner so that the following features are provided : ( 1 ) a gripping action of the surface of the heart during the massage ; and ( 2 ) a sucking effect on the surface of the heart so as to allow the heart to expand outward as the massager is moved or lifted upward ; thereby , enhancing the filling cycle of the chambers of the heart . in its operation , and in a general manner , the massager is inserted into the left chest via a small surgical incision . the massager has a heart - contacting member that is subcutaneously inserted through the incision and into the interior of the chest so that the now substernal massager may be placed on the anterior and lateral surfaces of the ventricular chambers of the heart . the massager allows the operator to manually compress the heart , which is confined by the pericardium , against the posterior chest wall structures . this manual compression results in an ejection of blood from the ventricular chambers similar to that occurring during normal cardiac contraction ( systole ). the massager is then relaxed so that the heart is allowed to fill passively ( diastole ). all of the embodiments of the present invention advantageously allow the heart to be compressed manually without the requirement of any external device , such as a pneumatic apparatus . as described herein , the massagers of the present invention have heart - contacting member having different forms but all forms of the heart - contacting members have a shape so that surface of the member contacting the heart is concave . this concaveness advantageously serves to conform the shape of the contacting surface of the massagers to the surface of the heart . each of the heart - contacting members has no dimension that is greater than three inches . as further described herein , the massagers of the present invention are subjected to a downward - axial force that most commonly occurs because the patient , who is receiving the heart massage , is commonly in the lying - down position . the massagers of the present invention may be subjected to forces in any direction necessary so as to accommodate any of the possible positions of the patient receiving the heart massage . still further , as described herein , some of the massagers of the present invention have a telescopic arrangement , wherein tube members move in an axial direction relative to each other or move in an axial direction in unison . as shown herein , the concavity of the heart - contacting member may take the shape of a cup . the cup member has the appearance of a plunger except that its portion of the plunger contacting the heart is closed . the cup has one end which is open and receives a handle and another end which is closed by a diaphragm having a concave shape for contacting the heart and a predetermined diameter . the ends are joined by a flexible sidewall which flares outward from the handle . one of the embodiments of the heart massager may be further described with reference to fig1 . the figure illustrates a massager 10 preferably having a grip 12 , and comprised of a handle 14 , a heart - contacting member 16 and , preferably , cushioning means 18 and padding means 20 . the heart - contacting member 16 is attached to one end of the handle 14 and has a surface 16a , shown in fig2 which is at least partially concave for contacting the heart . the heart - contacting member , in the form shown in fig1 and 2 , is a concave plate and has edges 16b and 16c . the embodiment 10 has some of the features of the well - known &# 34 ; richardson retractor &# 34 ; commonly used to hold back or retract elements of the abdominal wall to facilitate exposure of this wall during abdominal operations . the retraction of the abdominal wall is achieved by pulling on the handle of the richardson retractor which has a handle similar to the present invention . however , the bottom portion of the richardson retractor has a hoe - like retracting blade , whereas the present invention has a smoothly , contoured concave plate 16 having means to allow it to conform to the surface of the heart during its usage , and does not have any of the blade - like features of the richardson retractor . furthermore , the richardson retractor is used as a pulling device , whereas the present invention is used as a pushing device for pushing against the heart . further details of the massager 10 may be described with reference to fig2 which is a view taken along line 2 -- 2 of fig1 . the cushioning means 18 provides a cushioning action for the heart as the handle 14 is typically pressed downward . the cushioning means 18 is contoured so as to match the shape of the heart which the massager contacts . the cushioning means 18 , in cooperation with the concave plate 16 , provides a massager that conforms to the surface of the heart being massaged . the handle 14 and the heart - contacting member 16 are both preferably of a plastic material to facilitate electrodes and wiring therefor , along with other electrical functions ( e . g ., defibrillation , cardiac pacing and ekg ). the cushioning means 18 comprises deformable material having a portion 18a that conforms to the shape of the surface 16a of the concave heart - contacting member 16 , and a portion 18b which generally conforms to the shape of the anterior surface of the heart . the heart - contacting member 16 has edges 16b and 16c which are spaced apart from each other by a distance 16d in a manner corresponding to a typical concave shape , such as that of heart - contacting member 16 shown in fig2 . the portion 18b and , preferably , at least the edges 16b and 16c of the heart - contacting member 16 are covered with a padded material 20 , preferably a foam rubber or plastic in which two electrodes are embedded and have wiring routed through handle 14 . if desired , this material 20 may be formed as a part of cushioning means 18 . the pad 20 has an uneven , non - smooth surface that provides a slight gripping action which assists the external mating surface of the massager 10 to remain in contact , during the heart massage , with only one region of the heart . this assistance is desired so that once the massager is placed onto the desired location ( to be described ) of the heart , it remains at this location even in spite of the upward - downward movements of the handle 14 normally occurring during the massage of the heart . the embodiment 10 of fig2 as well those massagers of the fig3 - 8 have an overall dimension such as that defined by 16d , shown in fig2 that is measured from the tip of each of the edges 16b and 16c , and is of a value of less than about three ( 3 ) inches . as will be described , this three inch dimension is also pertinent to the diameter of the heart - contacting members having a cup shape , shown in fig3 - 8 . as will be further discussed with regard to the embodiment of fig9 - 11 , this three inch dimension may be further reduced to be less than about one inch . another embodiment 22 of a substernal heart massager , having shock absorbing or damping means for absorbing energy of sudden impulses that might accidently be applied to the massager 22 and might otherwise damage or scar the heart being massaged , is disclosed in fig3 . the massager 22 of fig3 as well as the massagers of fig4 and 5 , comprises the heart - contacting member 16 , cushioning member 18 and pad 20 all of which have already described with reference to fig2 . the apparatus 22 of fig3 also includes various elements having reference numbers all of which are given in the below table 1 . table 1______________________________________apparatus 22referenceno . element______________________________________24 first telescopic tube26 second telescopic tube28 guide slot formed in tube 2430 adjustment screw localed in slot 28 and af - fixed to tube 2632 compressible material comprising the shock absorbing means of massager 2234 electrode for transmitting electrical pulses or receiving electrocardiographic impulse36 electrode for transmitting electrical pulses or receiving electrocardiographic impulses38 opening in foot member 1640 conductor connected to electrode 3442 conductor connected to electrode 3644 grommet abutting between conductor 34 and opening 3846 grommet abutting between conductor 36 and opening 3848 plug for accepting signals from a cardiac stimulator for defibrillation / cardiac pacing______________________________________ the upper telescopic tube 24 has one of its ends connected to grip 12 and its other end movable within one end of the tube 26 which , in turn , has its other end connected to the heart - contacting member 16 . the tube 24 has a predetermined diameter that allows for movement in an axial direction relative to tube 26 . the guide slot and adjusting screw 30 provide the means for limiting the distance of relative motion ( axial movement ), in both the upward and downward directions , between the tube 24 and tube 26 . more particularly , the positioning or tightening of the screw 30 within the slot 28 of tubular member 24 allows for axial movement of tube 24 relative to tube 26 , until screw 30 engages the upper end of guide slot 28 . the amount of relative movement is determined by the axial - lengthwise dimension of guide slot 28 . during the downward movement , the compressible material 32 acts as a shock absorber to absorb the energy of any sudden downward - axially forces that may be imparted to tubes 24 and 26 . this shock absorbing means 32 , preferably of a spongy material , absorbs this unwanted sudden force that might scar , irritate or otherwise damage the heart organ . the spongy material 32 absorbs or dampens the accidental forces that might otherwise be imparted to the heart organ . the massager 22 further comprises electrodes 34 and 36 that extend out of and exposed by padded member 20 . the electrodes 34 and 36 are routed to a cardiac stimulator ( not shown ), commonly referred to as defibrillator , by means of conductors 40 and 42 , respectively , that pass through the handle and out the top of grip 12 . the cardiac simulator may also have provisions for cardiac pacing to be described hereinafter . grommets 44 and 46 respectively separate conductors 38 and 40 from the heart - contacting member 16 . in operation , the defibrillator ( not shown ) supplies electrical pulses , by way of an external conductor ( not shown ) and plug 48 , to the electrodes 34 and 36 which are in contact with the heart tissue , so as to terminate the ventricular fibrillation which most commonly causes cessation of spontaneous cardiac pumping action . alternately , the electrodes 34 and 36 may be arranged to serve as sensors for receiving electrocardiographic impulses and converting such impulses into appropriate electrical signals for transmittal to cardiac pacing equipment . another embodiment shown in fig4 as massager 52 has means for releasably engaging tubes forming a telescopic handle allowing for unison movement of the tubes in their engaged condition . the releasably engaging means further comprises resilient biasing means for releasing the engagement of tubes when the force applied to the tubes exceeds a predetermined limit . an indicator is also preferably provided to notify the operator that this force limit has been exceeded . the massager 52 has an actuator assembly 54 in the form of a telescopic arrangement . the telescopic assembly 54 a first tube 56 having one end connected to the grip 12 and its other end having dimensions selected to allow movement , in an axial direction , relative to a second tube 58 which , in turn , has its other end connected to the heart - contacting member 16 . further details of the massager 52 may be described with reference to fig5 which is a view taken along line 5 -- 5 of fig4 . fig5 shows an assembly 60 that serves as the means for releasing engaging the first and second tubes 54 and 56 , respectively , and also provides the resilient biasing means for releasing the engagement of the tubes when the force applied to the tubes exceeds a predetermined limit . the forces are applied , usually in a vertical - downward direction , to the grip and , thus to the handle comprised of both telescopic tubes . it is preferred , that an indicator be provided to warn the operator when the limit has been exceeded . the assembly 60 is comprised of elements having reference numbers all of which are given in the below table 2 . table 2______________________________________assembly 60referenceno . element______________________________________62 groove in sleeve 5864 ball66 spring means68 screw means70 contact switch72 excessive pressure warning indicator______________________________________ in operation , screw 68 is adjusted so as to establish the degree of force that pushes the ball 64 against the wall of groove 62 . the ball - groove contact provides the engagement between the tubes for the engagement between the tubes 56 and 58 allowing them to move in unison in an axial direction . this spring adjustment also determines the amount of force needed to cause the ball 64 to move out of the groove 62 which , in turn , allows the tube 56 to surge downward in a vertical direction relative to tube 58 . the operator , upon sensing such a surge , responds by removing or relieving the downward force applied to the massager 52 by way of grip 12 , thereby , avoiding any possible damage to the heart through excessive application of force . rather than rely on the operator &# 39 ; s perception of the movement of tube 56 relative to tube 58 , it is preferred to have the movement of ball 64 out of the groove 62 activate a switch 70 located below the groove 62 which , in turn , activates an excessive pressure warning indicator 72 . this warning mechanism may also be provided by having a switch 70a , the same as switch 70 , located on tube 56 and above tube 58 . the switch 70a only comes into contact with tube 58 when the ball 64 moves out of the groove 62 . the alternate arrangement of switch 70a allows tube 56 and tube 58 , both of actuator assembly 54 , to be more easily separated during the adjustment procedure of the spring 66 - screw 68 arrangement . a further massager 74 of the present invention , having inflatable / deflatable characteristics and the appearance of a plunger device , is shown in fig6 . the massager 74 has a handle 76 in the form of a tube with the grip 12 affixed to its upper end , first conduit means 78 which communicates with a source of positive fluid ( liquid or gas ) pressure and is affixed to handle 76 , second conduit means 80 connected to a source of negative fluid ( fluid or gas ) pressure and affixed to the handle 76 , and a heart - contacting member 82 having a flexible sidewall 84 and one end 86 that generally conforms to the surface of the heart . each of the conduit means 78 and 80 is in the form of a flexible tubular member . the heart - contacting member 82 comprises a cup having one end which is open and receives the handle 76 and another end which is closed by a diaphragm 86 having a concave shape for contacting the heart and a predetermined diameter . the ends of the cup 84 are joined by the flexible sidewall 84 which flares outward from said handle 76 . the source of positive fluid pressure used for the massager 74 may be in the form of a cartridge preferably containing co 2 or liquid water both under pressure , but may alternatively contain pressurized air . the source of negative fluid pressure used for the massager 74 may be a syringe or some other suction - generating source . the massager 74 may be further described with reference to fig7 which is a view taken along line 7 -- 7 of fig6 . the handle 76 has an opening 76a at one end that allows for the ingress , shown by arrow 88 , and egress , shown by arrow 90 , of fluid entering into and exiting out ( respectively ) of the cup member 82 . the ingress and egress of such fluid is controlled by fluid control means 92 comprising an assembly of elements having reference numbers all shown in the below table 3 . table 3______________________________________control assembly 92referenceno . element______________________________________94 control knob96 needle member98 guide means for needle member 96100 l - shaped conduit102 sealing member______________________________________ in operation , the control means 92 either lets a fluid medium , such as the medium co 2 , air raised to a positive pressure or liquid water under pressure , into the cup member 82 , or allows for the escape of this medium from the cup member 82 back to the source of positive liquid pressure . the manual control of this ingress or egress of the fluid medium is accomplished by means of the knob 94 opening or closing a valve ( not shown ) in l - shaped conduit 100 . the knob 94 may be arranged so that its depression causes the inward movement of needle 96 which , in turn , engages a valve seated in an opposing wall of the l - shaped conduit 100 , so as to allow the medium under positive pressure , shown by arrow 88 entering into the first conduit means 78 , to pass through the l - shaped conduit 100 near the top of the handle 76 and into the cup 82 , thereby , causing the laterally flexible wall 84 of cup 82 to expand outward and into its operative state ready to be used in the performance of the heart massage . in such an expanded state , the sealing member 102 , preferably of a silicone material in its hardened state , prevents the medium raised to a positive pressure in the cup 82 , from passing upward from control means 92 . sealing member 104 , preferably of a silicone material , prevents the medium , raised to the positive pressure , within cup 82 from finding its way out through the upper portion 84a of the flexible cup sidewall 84 . when it is desired that the cup member 82 be placed into its non - operable state , for insertion or removal from chest , the source of positive fluid pressure is removed from the first conduit means 78 , and the knob 94 is depressed causing the needle member 96 to move outward and thereby open the valve in member 100 . this allows the medium ( co 2 , liquid water or air raised to a positive pressure ) trapped in the cup 82 to find its way out of the cup 82 by way of the l - shaped member 100 , in the direction as shown by arrow 90 . negative fluid pressure acting as a suction source , may be applied to the cup 82 by way of the second conduit means 80 that has a tube or channel running into the open end 76a of handle 76 . the second conduit means 80 may further comprise control means ( not shown ) for selectively activating and deactivating the negative pressure entering handle 76 , but such a pressure - selective control device may also be arranged in the negative fluid pressure source itself . the suction effect provided by means 80 , causes the concave - conforming surface of diaphragm 86 of the cup 82 to more effectively grip onto the heart allowing for enhanced pumping of blood through the heart during the cpr method of the present invention to be described hereinafter . a further massager embodiment 106 , partially shown in fig8 is designed to also assist in the filling process ( diastole ) of the heart that occurs during the heart massage . massager 106 has a handle 108 having within its hollow a suction channel 110 similar to the suction channel of device 80 of fig7 . the handle 108 of fig8 has a heart - contacting member 112 in the shape of a cup connected to its open end 108a and sealed thereto by sealing means 114 preferably of a silicone material . the heart - contacting member 112 serves the same function as the heart - contacting member 16 having the form of a concave plate of fig1 and has a cup - like shape and a structure very similar to the cup - like heart - contacting member 82 of fig6 and 7 . the heart - contacting member 112 has a continuous sidewall 116 , and one end 118 with a predetermined diameter and its other end connected to said opened end 108a of the tubular handle 108 . the sidewall 116 expands outward when subjected to an applied downward force . the end 118 has an outer surface with a plurality of circular shaped indentations 120 comprising suction - cups members . the outer surface has this concave shape so as to be adapted for contacting the heart . in operation , the suction cups 120 , in cooperating with the suction channel 110 connected ( not shown ) to a source of negative fluid pressure , provide a sucking or gripping effect for the massager 106 onto the surface of the pericardium , so that the muscle of the heart along with its blood controlling chambers are expanded outward upon the upward movement of the handle 108 of the massager 106 . this upward movement of handle 108 expands the heart muscle so as to enhance the filling of its heart chambers during the relaxation period of the cpr process to be described . another embodiment of the present invention , having umbrella - like features and allowing for its insertion through the chest by way of a surgical incision of less than about one inch , is shown in fig9 as massager 122 . the umbrella - like massager 122 has a handle 124 , in the form of a telescopic arrangement , and a concave cup member 126 which is compressible and radially expandable in response to a force applied to its outer surface . in general , the handle 124 comprises first and second telescoping tubes for axial movement relative to each other , one of the tubes being connected to the grip 12 . the first and second tubes are interconnected by a rod with a carriage member on an end thereof and displaceable within the tubes . the carriage member has affixed thereto a plurality of flexible spoke members . the flexible spoke members each having one end affixed and circumferentially spaced apart from each other in the carriage member . each of the flexible spoke members further has its other end axially extending out of the carriage member . the collapsible cup comprises stretchable material and has flexible sidewall having means for respectively capturing and holding each of the other ends of the flexible spoke members axially extending out of the carriage member . more particularly , the collapsible cup 126 has a plurality of flexible ribs 128 , a plurality of flexible body portions 130 , and , preferably , a cushion member 132 that extends around the lower rim portion of cup 126 as seen in fig9 . the massager 122 further comprises sensors / electrodes 134 and 136 located on the circumference of the cushion member 132 and which are respectively routed to plug 138 via conductors 140 and 142 for connection to a defibrillator or cardiac simulator . the operation of the sensors / electrodes will be described below , in connection with the description of fig1 . the massager 122 preferably has third conduit means in the form of a flexible tubular member that extends into a hollow of the handle 124 and which is connected ( not shown ) to a source of negative fluid pressure to provide the suction effect of the heart muscle in a manner as described with reference to conduit means 80 and 110 of fig7 and 8 respectively . the massager 122 may be further described with reference to fig1 , which is a view taken along line 10 -- 10 of fig9 . fig1 , in addition to those elements shown in fig9 illustrates a plurality of elements given in the below table 4 along with their respective reference number . table 4______________________________________massager 122referenceno . elements______________________________________146 first tube member of telescopic handle 124148 second tube member of telescopic handle 124 - 150 central pole interconnecting tubes 146 and 148152 attaching means for pole 150154 carriage member for riding and being dis - placed within tube 146156 plurality of flexible spoke members158 outer surface of a piece of stretchable material having a cup - like shape in its expanded state160 inner surface of the stretchable flexible material having a cup - like shape in its expanded state162 bottom concave surface of the cup - like flexible material______________________________________ the tube 146 has a bore with a predetermined dimensions , and an exit portion . as best illustrated in the upper portion of fig1 , the tube 148 has a predetermined outer dimension selected to allow the tube 148 to move within the bore of the tube 146 in a reciprocating manner . the tube 148 is connected ( not shown ) to the grip 12 . the central pole 150 has one end affixed to the inner diameter of the tube 148 by attachment means 152 and has its other end glided within the bore of the tube 146 by carriage member 154 . the carriage member 154 rides and is displaced within the tube 146 . the attachment means 152 may be of any suitable form such as welding so long as the pole 150 remains fixed to the tube 148 during the reciprocating movement of tube 148 . the carriage member 154 has affixed therein a plurality of flexible spokes 156 , each being circumferentially spaced apart from each other and each vertically extending downward from the carriage member 154 . the compressible and radially expandable cup member 126 comprises a stretchable material and is connected to the carriage member 154 by means of the plurality of spoke members 156 that are lodged and held in each of the ribs 128 of the expandable material by appropriate capturing means affixed to the expandable material . as best seen in the lower portion of fig1 , the ribs 128 are bowed outward by the action of their captured radially - expanding spokes 156 . in operation , the massager 122 operates in a similar manner as a conventional umbrella allowing the stretchable material to have a cup - like shape in its expanded condition and a tubular - like shape in its compressed condition . when grip 12 is retracted upward relative to tube 146 , the carriage member 154 moves upward , as viewed from fig1 , which correspondingly causes the spoke members 156 to be withdrawn upward which , in turn , results in the ribs 128 being pulled radially inward until all the cup - like shaped material is compressed and drawn into the tube 146 . such a compressed condition is shown in fig1 , wherein it is seen that spoke members 156 , in their relaxed condition , merge at a central region 164 and taper outward therefrom toward both their upper and lower regions . this compressed condition is used for the insertion and withdrawal of the massager 122 from the chest cavity . with further reference to fig1 , the maximum outer diameter of the cushion member 132 , shown as dimension line 132a , is preferably less than about one inch . such a dimension allows the umbrella massager to be first inserted into a surgical incision of less than one inch for later intra - thoracic positioning of the massager to accomplish the massage of the heart muscle . the cup member 126 is closed off by the bottom wall 162 which has a flexible diaphragm - like concave shape to adapt to the surface of the heart . when it is desired to cause the cup member 126 of fig9 - 11 to radially expand outward from its compressed condition so as to obtain its operative state , the grip 12 is gripped by the operator and a force is applied so that the grip moves toward the exit portion of tube 146 . as viewed from fig1 , this movement in turn causes the carriage member 154 to move downward which , in turn , causes or allows the spoke members 156 to move the ribs radially outward until all of the cup - like shaped material of cup 126 is expanded outside of the tube 146 . following its use , the umbrella massager 122 is retracted into its tube 146 for removal from the chest cavity . a still further embodiment , massager 166 , of the present invention is shown in fig1 having provisions for receiving signals transmitted from a plurality of external devices . the massager 166 has a handle 168 to which is mated , at a slight angle , a collar 170 of an endoscope 172 that accepts a cable 174 routed from an endoscopic light source 176 . the endoscope 172 has a cable 178 that runs downward into the lower portion of handle 168 to direct the related light thereto and is used by the operator of endoscope for viewing purposes . the endoscope may be any of the known commercially available endoscopes . the massager 166 has sensors / electrodes 134 and 136 in the heart - contacting member 126 having a cup like shape and that contacts the heart during massage . in one form , that is , the sensors 134 and 136 act as elements for receiving signals such as impulses generated by the heart , whereas in another form that is , electrodes transmit electrical pulses received from a defibrillator . the sensors / electrodes are connected by internal cabling ( not shown ) that exits handle 12 as cabling 184 which , in turn , is connected to a switching device 186 . the device 186 selectively interconnects the massager 166 to one or more of the external devices given as follows : ( 1 ) electrocardiograph 188 ; ( 2 ) cardiac pacing device 190 ; ( 3 ) defibrillator 192 ; and ( 4 ) excessive compression pressure warning light 72 . each of the electrocardiograph 188 , the cardiac pacing device 190 and the defibrillator 192 may be of a known commercially available type . the mechanism that controls the warning light 72 is not shown in fig1 , but is shown and has been described with reference to fig5 . the devices 188 , 190 , 192 and 72 assist or enhance the operation of the massage of the heart and are respectively interconnected to switch 186 by means of cables 194 , 196 , 198 and 200 . in operation , the endoscopic device 172 allows the massager 166 to be guided in a very accurate manner to its desired position of the heart . the device 172 allows the operator to see the subcutaneous movement of the concave - cup 126 , serving as the foot of the massager 166 , so that the foot may be exactly placed onto its desired location on the heart during the heart massage . the selection of device 188 by switch 186 allows the electrocardiogram ( ekg ) to record the changes in the electrical potential caused by the heartbeat . such changes may be used to determine and correct for any abnormal electrocardiograph rhythms that may be present during the cpr procedure involved in the cardiac arrest . the selection of device 190 by switch 186 may be used to synchronize and stabilize the ekg signals created by the heartbeats during the cardiac arrest condition . the stabilization provided by device 190 may assist in a more rapid recovery from a cardiac arrest condition . the selection of device 192 by switch 186 may be used , as previously discussed , for defibrillation of the heart muscle to allow it to return to its normal operating condition in a more rapid manner . another instrument ( not shown ) that finds particular usage with the practice of the present invention and that is external from the massager 166 , is a blood pressure monitor unit that normally contacts the patient by means of a cuff device . this monitor is used to detect the increased flow of blood generated by the cpr techniques of the present invention relative to the closed cpr techniques of the prior art . this elevated blood pressure may be used as a tool or indicator that a hemodynamically significant compression of the heart organ is actually taking place during the internal cpr , to be described , using the present invention . the blood pressure may be used a monitoring means to advantageously place , maintain or move the massagers of the present invention to or from desired positions along the heart , so that the compression thereof causes the advantageous enhancement of the blood flow as manifested by elevated blood pressure during the internal cpr procedure . it should now be appreciated that the practice of the present invention provides for various massagers all having special provisions for allowing for increased blood flow , while at the same time being of relatively small dimensions to minimize the surgical incision necessary for the internal cpr technique of the present invention . method and technique for internal cardio - pulmonary resuscitation ( cpr ) of the present invention the method of the present invention may be described with reference to fig1 - 18 in which fig1 and 14 illustrate the location of the anatomical elements related to the present invention ; fig1 and 16 illustrate the predetermined location of surgical incision of the present invention ; fig1 , composed of fig1 a and 17b , illustrates the anterior - posterior compression of the heart achieved according to the practice of the present invention ; and fig1 , composed of fig1 a , 18b and 18c , illustrates the lateral - medial compression of the heart achieved according to the practice of the present invention . fig1 is a right lateral view of the thorax ( chest ) 210 showing the human heart 212 located in its lower portion . fig1 further illustrates the location , relative to the heart 212 , of the phrenic nerve 214 , the ribs 216 ( the plurality of ribs being indicated by a single reference number 216 ), the esophagus 218 , vertebrae 220 , azygous vein 222 and diaphragm 224 . further details of the heart 212 are shown in fig1 which is a cross - sectional view of the thorax . the heart 212 is enclosed in a tough , fibrous sac called the pericardium 226 , which is attached by fibrous strands to the posterior portion of the sternum 228 . the heart 212 has right and left cardiac ventricles 230 and 232 , respectively , which propel blood into the heart and the body , commonly called systemic circulation . as shown in fig1 , behind the heart 212 , overlying the thoracic vertebrae 220 , are located the esophagus 218 and the descending aorta 234 ( the main artery carrying blood to the body from the heart ). the particular arrangement of these anatomical features lend themselves to the ability of the massagers of the present invention to provide the desired compression of the heart in both the anterior - posterior and the lateral - medial directions once these massagers are intra - thoracically and substernally positioned . the positioning of any of these massagers may be described with reference to fig1 showing the rib cage 236 of the body . fig1 primarily illustrates the left side of the rib cage 236 , as well as the location of the heart 212 ( shown in phantom ) relative to the left nipple 238 , sternum 228 ( shown in phantom ) and the fourth intercostal space 240 . as shown in fig1 , a small surgical incision 242 is made in the fourth intercostal space 240 between the fourth and fifth ribs , approximately 1 - 3 inches in length . this incision 242 is created on the anterior chest wall along a line between the left nipple 238 and the lateral border 244 of the sternum 228 . this 1 - 3 inch incision is applicable for the use of the massages illustrated in fig1 - 8 and 12 . the massagers of fig9 - 11 require only an about one ( 1 ) inch incision . unlike the known prior art methods such as that of the &# 39 ; 932 patent discussed in the &# 34 ; background &# 34 ; section above , the surgical incision 242 of the present invention does not enter the upper abdomen beneath the xiphoid process of the sternum , but is made in the left chest over the ventricles of the heart . after such an incision is made , a sharp surgical instrument is used to provide sharp dissection , preferably in the fourth intercostal space , thereby , allowing for the entrance of the finger of an operator which is used to locate , by finger palpation , the apex region of the heart . the massager , such as massager 10 of fig1 having the heart - contacting member with a surface which is at least partially concave for contacting the heart 16 , may now be introduced onto the heart and such introduction may be described with reference to fig1 . fig1 shows the massager 10 introduced through the ribs which are shown as 216 2 , 216 3 , 216 4 , 216 5 and 216 6 with the subscript indicative of the particular rib of the body . the massager is inserted between the fourth 216 4 and fifth 216 5 ribs and onto the apex region 246 of the heart 212 . the incision provides for the entrance of any of the massagers of the present invention between the ribs and allows such massager to be placed on the heart muscle without incising the pericardium . cardiac compression is then carried out by manual depression of the introduced massager . the compression of the heart may be described with reference to fig1 and 18 . fig1 a shows the right and left ventricles 230 and 232 as being divided by septum 248 . fig1 a further shows heart 212 in its non - compressed state , whereas fig1 b shows heart 212 in its compressed state created by a pressure applied to the massager 10 . the compression is predominantly in the anterior - posterior direction 250 , thereby compressing , as shown in fig1 b , both the right and left ventricles 230 and 232 . however , the manual stroke also has a lateral - medial component , which is described with reference to fig1 . fig1 is composed of fig1 a , 18b and 18c which respectively illustrate the esophagus 218 and aorta 234 located between the vertebrae 220 and the heart 212 shown in its non - compressed condition , an initial compressed condition of the heart 212 , and a final compressed condition of the heart 212 . as seen in fig1 a , the esophagus 208 and aorta 234 are in their non - restricted state when the heart 212 is in its non - compressed condition . fig1 b illustrates the initial contact of massager 10 with the heart 212 . the massager 10 is shown in fig1 b in an enlarged manner , relative to that of fig1 and 17b , so as to more clearly illustrate the conformity of the heart - contacting member of the massager 10 to the surface of the heart . as seen in fig1 b , when the massager 10 is pressed downward , a lateral - medial component 252 is applied to the heart causing the heart to compress the ventricles 230 and 232 against the vertebrae 220 . the final compressed state of ventricles 230 and 232 is shown in fig1 c . fig1 c shows the heart with the massager removed so as to more clearly illustrate the compressed state of the ventricles 230 and 232 , as well as the compressed state of the esophagus and , more particularly , the compressed or restricted state of the aorta 234 . because of the anatomic location of the aorta 234 , the compression stroke along the lateral - medial direction 256 also produces a partial occlusion of the aorta 234 which , in turn , as previously discussed , elevates the blood pressure to a greater degree in the upper portion of the body than in the lower part . this compression , as previously mentioned , selectively increases the blood flow of the heart and the brain . this selective increase in blood pressure and flow does not occur in conventional open - chest or closed - chest manual cpr techniques . further , this increased blood flow , which is a significant advantage of the present invention , has never been reported during the prior art open - chest or closed chest manual cpr techniques . in the practice of the present invention , since the pericardium is left intact , ( unlike open bi - manual cpr ) the pericardial sac holds the heart in a favorable position during compression and prevents dilation of the heart chambers . these factors may also improve resuscitation using the present invention . it should now be appreciated that the method of the present invention provides an internal , manual cpr technique that increases the blood flow in the vital organs improving the probability of patient survival . results achieved by the heart massager and method both of the present invention a massager of the present invention was introduced using minimum invasive surgery directly onto the heart of ten separate swines for the performance of direct cpr in a manner as previously described hereinbefore . the results of the direct cpr massage of the ten swines of the present invention , herein termed minimally - invasive ( mid ) cpr , were compared against the results obtained from massaging the hearts of the ten swines using open chest ( oc ) cpr . the parameters monitored during both the mid cpr and oc - cpr massagers are given in the below table 5 . table 5______________________________________parameter units of measurements______________________________________cardiac index ( ci ) cc / kg / minarterial ( ph ) acidity / alkalinity on a scale from 0 to 14arterial lactate ( al ) mml / dlmixed venous po2 ( pvo2 ) mmhg______________________________________ for each swine , ventricular fibrillation was induced and the parameters of table 5 were recorded every ten minutes ( m ) in which five minutes of recording was allocated to the oc - cpr massage and the remaining five minutes of recording allocated to the mid cpr . the initial ( 0 minutes ) recording was assigned as the baseline ( base ) for each of massage techniques . the results of the testing are given in the below table 6 . table 6______________________________________time cpr ci ph al pv02______________________________________base oc 78 . 4 ± 27 . 9 7 . 42 ± 0 . 06 1 . 44 ± 0 . 48 53 . 8 ± 11 . 1mid 78 . 4 ± 11 . 8 7 . 42 ± 0 . 10 1 . 30 ± 0 . 35 53 . 4 ± 24 . 710 m oc 26 . 8 ± 4 . 5 7 . 34 ± 0 . 09 4 . 56 ± 0 . 43 23 . 4 ± 2 . 6mid 40 . 5 ± 9 . 5 7 . 31 ± 0 . 10 3 . 60 ± 1 . 24 24 . 2 ± 6 . 820 m oc 35 . 1 ± 12 . 4 7 . 31 ± 0 . 04 5 . 14 ± 0 . 70 25 . 8 ± 4 . 5mid 39 . 5 ± 11 . 9 7 . 30 ± 0 . 10 4 . 56 ± 1 . 62 22 . 5 ± 8 . 930 m oc 26 . 7 ± 10 . 5 7 . 23 ± 0 . 10 5 . 86 ± 0 . 62 24 . 8 ± 6 . 0mid 43 . 9 ± 5 . 5 7 . 26 ± 0 . 10 5 . 04 ± 1 . 80 23 . 7 ± 9 . 0______________________________________ the values ci , ph , al and pvo2 are expressed in the mean values obtained (±) standard deviation from these mean values . from table 6 it can be seen that all of the parameters measured during the method of the present invention ( mid ) were comparable or better than those obtained from the open chest ( oc ) cpr method . it should now be appreciated that the practice of the present invention provides for an apparatus and a method for a heart massager that yields results that are comparable and even better than that obtained from the open - chest cpr requiring major cardiac thoracotomy . the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof and , accordingly , reference should be made to the appended claims , rather than to the foregoing specification , as indicating the scope of the invention .

a heart massager for substernal heart massage is disclosed . the heart massager has a heart - contacting member having a surface which is at least partially concave for contacting the heart , cushioning on the surface of the contact member , and a handle attached to the heart - contacting member for manually manipulating the massager . the partially concave surface allows the heart - contacting member to conform to the shape of the heart . the handle is substantially upright with respect to the surface of the heart - contacting member . the handle is significantly offset from a central portion of the heart - contacting member in the region where it attaches to the member . the cushioning covers substantially the entire surface of the heart - contacting member so as to form a solid surface .

referring now to fig1 of the drawings , there is shown a bracelet 1 in the form of an elongated relatively narrow web of a durable light - weight plastic material , suitable for molding , such as polyethylene or polyurethane . a web portion 3 , comprising the major length of the bracelet 1 , preferably has a width of approximately 0 . 5 inches and a thickness in the range of from 0 . 06 to 0 . 25 inches . at one end of the bracelet 1 there is an outward taper 5 terminating in a widened substantially rectangular stub 7 having a substantially rectangular aperture 9 . the corners of the rectangular aperture 9 are rounded as are the two extreme exterior corners of the stub 7 . the length of the aperture 9 is parallel to the width of the web 3 , i . e ., transverse to its longitudinal axis , and is equal to or just slightly greater than the width of the web 3 . that is , where the width of the web 3 is 0 . 5 inches , the length of the aperture 9 would preferably be in the range of 0 . 5 to 0 . 7 inches . the radius of curvature of each corner of the aperture 9 is approximately 0 . 08 inches . the width of the aperture 9 along a dimension parallel to the longitudinal axis of the web portion 3 and transverse to the width of the web portion 3 is less than its length but greater than the thickness of the bracelet web portion 3 . in the preferred embodiment where the width of the web 3 is 0 . 5 inches , the width of the aperture 9 would preferably be in the range of 0 . 3 to 0 . 4 inches for a bracelet having a web thickness of up to 0 . 25 inches and 0 . 1 to 0 . 4 inches for a web thickness of as little as 0 . 06 inches . at the end of the bracelet 1 opposite the end having the stub 7 is a connector portion 11 defined by a step in the width of the web 3 forming a neck 13 followed by a semicircular barb 15 , the width of the barb 15 in a direction transverse to the longitudinal axis of the web 3 being substantially equal to the width of the web 3 , and the thickness of the neck 13 and barb 15 being substantially equal to the thickness of the web 3 whereby each of the web 3 , neck 13 , barb 15 and stub 7 has a rectangular cross section transverse to the longitudinal axis of the web 3 . the end - to - end length of the bracelet 1 measured from an apex of the barb 15 to a distal edge of the stub 7 is 111 / 8 inches in the preferred embodiment of the invention . although this dimension will be suitable for most wrists , variations may be made within the scope of the invention to accommodate smaller or larger appendages . evenly spaced along the longitudinal axis of the web 3 and spanning approximately two thirds of the length of the web portion 3 , beginning proximate the outward taper 5 of the bracelet 3 , are a series of like - dimensioned , evenly spaced , rectangular apertures 17 having rounded corners which serve as locking holes for the barb 15 . the width of each of the apertures 17 in a direction parallel to the width of the web 3 and transverse to the longitudinal axis of the web 3 is preferably equal to or slightly less than one half the width of the bracelet web portion 3 , and its length in a direction parallel to the longitudinal axis of the web 3 is slightly greater than the width of the barb 15 , e . g ., in the preferred embodiment of the invention with a barb 15 having a width of 0 . 5 inches , the web 3 has apertures 17 with a width of 0 . 241 inches and a length of 0 . 6 inches . the radius of curvature of each corner of each aperture 17 is approximately 0 . 08 inches . the spacing between the apertures 17 will depend on the thickness of the web portion 3 , the greater the thickness , the closer the spacing permitted . in the preferred embodiment of the invention , with a web portion 3 having a width of 0 . 5 inches and a thickness of approximately 0 . 125 inches , the spacing between adjacent extremities of the apertures 17 is preferably on the order of 0 . 375 inches . referring now to fig2 - 5 , in use , the bracelet 1 is wrapped about the wrist ( or ankle ) of a person and the barb 15 is inserted through the rectangular aperture 9 in the stub 7 and pulled through until a snug enough fit is obtained to prevent movement of the bracelet 1 relative to the wrist or , if a loose fit is desired , to prevent the bracelet from sliding over the hand of the wearer . the barb 15 is then twisted approximately 90 degrees , as best seen in fig3 to align its width with the length of the nearest aperture 17 through which the barb 15 is then inserted . the barb 15 is then released thereby permitting the resilience of the plastic material from which the bracelet 1 is molded to restore the barb to its original orientation as best seen in fig5 . the barb 15 is thereby prevented from being withdrawn through the aperture 17 and the neck 13 is captured within the aperture 17 . to remove the bracelet , the wearer need only twist the barb 15 again to align it with the aperture 17 at which time the barb 15 can be passed back through the aperture 17 and then withdrawn through the aperture 9 in the stub 7 . depending on the size of the hand of the wearer , it may be possible to slide the bracelet over the hand by partially withdrawing the web portion 3 through the stub 7 without causing the barb 15 to pass through the stub aperture 9 . it is to be appreciated that the foregoing is a description of preferred embodiment of the invention to which variations and modifications may be made without departing from the spirit and cope of the invention .

a bracelet has a guide aperture at one end , a barb at the opposite end , and elongated holes running along its length . the barbed end can be pulled through the guide aperture and then twisted and inserted into an adjacent hole for being captured therein . the holes in the bracelet have rounded corners to enhance the flow of the polymeric material from which the bracelet is formed during an injection molding process and to alleviate the formation of stress points which otherwise occur at the corners of rectangular holes .

the present invention will be further described in the following examples and comparative examples in which compositions demonstrating various facets of the invention were prepared . the examples are by way of illustration only , and are not to limit the invention . table 1 lists the sources of the fats used in the examples and comparative examples . the effect of adding 20 % of a fat rich in sss triglycerides ( the stearin fraction of palm oil ) or 20 % of a fat rich in ssu ( the mid fraction of palm oil ) on the solids content at 5 ° c . of a fat rich in suu ( the double fractionated olein fraction of palm oil ) was determined using the method described above . the safa contents , the measured solids contents and the solid : safa ratios are given in table 2 . adding a fat rich in ssu produced the same increase in solid fat at 5 ° c . as adding a fat rich in sss , but with a smaller increase in the amount of safa i . e . to 45 % rather than to 49 %. this data shows that ssu triglycerides are more effective than sss at increasing the solids : safa ratio of a suu rich fat . ice creams were prepared using fats and fat blends according to the present invention ( examples 1 - 11 ). these are based on one or more palm oil components , some of which are diluted with rapeseed oil or sunflower oil . ice creams were also prepared using fats and fat blends which are outside the scope of the invention ( comparative examples a - f ). the fats and fat blends are listed in table 3 . the triglyceride composition , safa content , and solid fat content of the examples and comparative examples are given in table 4 . the exact composition of fats varies between sources and the numbers given are typically averages . because of experimental and rounding errors , the sum sss + ssu + suu + uuu may not be exactly 100 % in every case . examples 1 - 10 according to the invention all have safa contents of no more than 55 %, sss & lt ; 8 % ( hence long chain sss & lt ; 8 %), and solid fat : safa & gt ; 1 . comparative examples a - f do not satisfy all of these criteria and are therefore outside the scope of the invention . ice creams were prepared to the formulation given in table 5 using the examples as the fat source , according to the following process . water was placed in a jacketed mix tank at 85 ° c ., then all the ingredients except for the colour and flavour were dissolved by mixing with a high shear mixer . hot water was circulated in the jacket of the tank to maintain a premix temperature of 65 ° c . the premix was heated using a plate heat exchanger to a temperature of 83 ° c ., homogenised using a the first stage of a crepaco double stage valve homogeniser at a pressure of 140 bar . after holding the mix at 82 to 85 ° c . for 12 seconds , the mix was cooled using a tubular heat exchanger to 4 - 6 ° c . and held at this temperature for at least 2 hours before freezing . the colour and flavour were added to the cooled mix . the mix was processed through a continuous ice cream freezer ( type apv technohoy mf75 ) fitted with an open dasher rotating at a speed of 370 to 400 rpm and operating with a barrel pressure in the range 1 - 4 bar . the ice creams were produced with a mix flow rate of between 0 . 6 and 0 . 63 litres / minute , with an overrun of 100 % and an extrusion temperature in the range − 7 . 5 to − 7 . 8 ° c . they were collected in 500 ml waxed paper cartons and then hardened in a blast freezer at − 35 ° c . for a period of 2 hours . the hardened ice creams were stored at − 25 ° c . examples 1 - 10 according to the invention all produced ice creams which processed well ( i . e . they could reliably be aerated to 100 % overrun and were smooth and dry on extrusion ) and had good shape retention . on eating , the ice creams had a good texture and did not have a waxy mouthfeel . comparative example a ( coconut oil ) is a conventional fat source for ice cream . the ice cream processed well and had good shape retention on extrusion . however the fat has a very high safa content ( 92 %). although the amount of sss triglycerides is high ( 76 %), the sss triglycerides in coconut oil almost entirely have short acyl chains , so that the amount of long chain sss triglycerides is less than 8 %. as a result , comparative example a did not have a waxy texture . comparative examples b ( palm oil ) and c ( a blend of palm oil and the stearin fraction of palm oil ) have somewhat lower safa contents , solid fat : safa & gt ; 1 , ssu : suu & gt ; 1 and ssu + suu & gt ; 50 %. however , unlike coconut oil , the sss triglycerides in palm oil and palm oil fractions are almost entirely long chain sss triglycerides ( approximately 99 % for po and psf ). as a result , the long chain sss triglyceride contents of comparative examples b and c are greater than 8 %, and although the ice cream made with these fats processed well , it had an unpleasant waxy texture . comparative examples d , e and f have low safa contents ( 36 to 40 %), but have solid fat : safa & lt ; 1 . comparative example d ( a blend of coconut oil and sunflower oil ) is largely comprised of sss and uuu triglycerides . it has ssu + suu & lt ; 50 % and also ssu : suu & lt ; 1 . its solid fat content was measured to be 18 %. on processing , aeration was observed to be inconsistent , and the ice cream had a thin , icy , cold eating texture . the poor quality of the ice cream arises from the low solid fat content . although this blend has a sss triglyceride content of about 22 %, these are almost entirely short chain triglycerides which do not cause a waxy mouthfeel . comparative example e ( the double fractionated olein fraction of palm oil ) has ssu + suu of 90 % and sss & lt ; 8 %. however the ssu : suu ratio is less than one and the solid fat content was only 2 . 5 %. ice cream made with this fat processed very poorly . it was not possible to achieve 100 % overrun . on extrusion , the ice cream was wet and non - uniform , and had poor shape retention . popping of air bubbles was observed , i . e . air was visibly lost from the structure . on eating , the ice cream had a thin , coarse , icy texture and was very cold - eating . again , the poor quality of the ice cream is due to the very low solid fat content . comparative example f ( a blend of palm oil and sunflower oil ) had solids : safa and ssu : suu ratios of 0 . 94 and 0 . 99 respectively . ice cream made with this fat also processed poorly and suffered from the same defects as comaparative example d . the comparative examples show that simply replacing some of a fat high in safa with one low in safa is not sufficient to produce good quality ice cream with a safa content of no more than 55 %. however , the examples according to the invention show that by carefully choosing the fat source according to certain specific criteria ( the solid fat content and triglyceride composition ), good quality ice cream can be produced with safa contents of less than 55 %, and even as low as 35 %.

a frozen aerated confection having an overrun of at least 40 % and a fat component in an amount of 2 to 20 % , said fat component comprising triglycerides of fatty acids wherein less than 70 % of the fatty acids in the triglycerides are saturated , less than 8 % of the triglycerides are sss triglycerides ; characterized in that the ratio of the percentage of fat that is solid at 5 ° c . to the percentage of the fatty acids in the triglycerides that are saturated is greater than 1 and in that the fat component comprises at most 60 % cocoa butter or shea nut oil .

in accordance with the present invention , the improved ultrasonic surgical tool provides enhanced tactile feedback to the surgeon and may be adjusted to customize the feedback , depending on the preference of the surgeon . additionally , the ultrasonic tool of the present invention can be configured to cut a wide variety of tissues by altering the blade structure alone , or in combination with the operating mode . it thus is understood that the general shape of the present surgical tool can be changed to be similar to any of the 75 plus conventional shapes available from several microtome / sharps manufacturers , which are specifically designed to cut specific tissue or material type , while still incorporating the advantages of the present invention . incorporation of the invention into known blade configurations will give a normally &# 34 ; static &# 34 ; blade / tool / sharps improved capabilities , as discussed below . the following discussion of the invention , however , will focus on an improved cutting tool configuration in combination with the other aspects of the present invention . as schematically shown in fig1 an ultrasonic surgical system 24 ultimately vibrates a surgical blade 26 . the blade 26 couples to an ultrasonic transducer ( not shown ) mounted in a handpiece 28 which is driven by a control system 30 . a surgeon grasps the handpiece 28 and manipulates the blade 26 within a patient . a cable 32 transmits the ultrasonic driving signals from the control system 30 to the transducer within the handpiece 28 . referring to fig2 a preferred embodiment of surgical blade 26 is shown . the blade 26 includes a cutting section 34 at its distal end . as seen best in fig4 the cross section of the cutting section 34 reveals a central channel or relief 36 formed into each side . the blade 26 is symmetric about a vertical plane through the center . the relief portion 36 allows the knife blade 26 to cut through various types of tissue with a minimum thermal footprint . the thermal footprint of a blade includes all the surfaces in contact with the tissue . the blade 26 vibrating at ultrasonic frequencies can produce a substantial amount of heat from the frictional and ultrasonic contact with the tissue . the size of the relief 36 , or percentage of area of the blade 26 out of contact with the tissue , directly affects the thermal footprint . adjacent the relief 36 , tissue contact surfaces 38 extend for a distance generally parallel to the plane of the blade 26 towards the edge of the blade 26 . in general , the width of each contact surface 38 in this plane is within the range of from about 0 . 0 to about 0 . 050 inches , and preferably within the range of from about 0 . 015 to about 0 . 025 inches . these contact surfaces 38 represent the widest portion of the blade 26 along an axis transverse to the plane of the blade and produce a substantial amount of thermal friction with the tissue . typically , the thickness of the blade through contact surface 38 is within the range of from about 0 . 010 to about 0 . 050 inches , and preferably within the range of from about 0 . 015 to about 0 . 025 inches . the size of the contact surfaces 38 also directly affects the thermal footprint . smaller contact surfaces 38 reduce the thermal footprint of the blade 26 . with reference to fig4 the sharpened edge 27 of the blade 26 comprises a first taper 40 which is separated from the contact surface 38 by a second taper 42 . both the first and second tapers widen in the medial direction . as illustrated in fig4 the first taper 40 is formed by the intersection of two inclined surfaces 41 . each inclined surface 41 tapers from a vertical central axis at a taper angle ranging between about 5 ° and about 45 °, and more preferably between about 15 ° and about 30 °. the taper angle most preferably equals about 30 °. consequently , the first taper 40 has an included angle ranging between about 10 ° and about 90 °, and preferably equals about 60 °. it has been determined that taper angles within the above ranges provide a good balance of mechanical sharpness and hemostatic tissue activity , as discussed below . it is also understood that the cross - sectional profile of the blade edge 27 can be changed to obtain a different balance of these effects . for instance , fig4 a illustrates an alternative cross - sectional shape of a first taper 43 . in this embodiment , a pair of intersecting curved surfaces 47 ( preferably concave surfaces ) form the first taper 43 , thus giving the first taper 43 a &# 34 ; hollow - ground &# 34 ; shape . each surface desirably has a radius of curvature ranging between about an eighth ( 1 / 8 ) to about ten ( 10 ) times the blade thickness , more preferably between about a quarter ( 1 / 4 ) to about five ( 5 ) times the blade thickness , and most preferably between about a half ( 1 / 2 ) to about two ( 2 ) times the blade thickness . the hollow - ground configuration of the first taper 43 also has a shorter depth 49 , as measured in the direction of the vertical central axis , in comparison to the cross - sectional profile of the blade edge 27 illustrated in fig4 . the short , hollow - ground profile provides for a sharper cutting edge at the tip with increased thermal properties directly behind the tip . of course , the smaller the radii of the curved surfaces , the closer these two effects ( sharpness and thermal hemostatic activity ) are brought together . with reference back to fig4 the second taper 42 desirably range between about 3 ° and about 45 °, and more preferably the taper is about 8 °. the angles of the tapered portions directly affect the character of cut and associated drag , or feel , experienced by the surgeon . a short taper , such as 45 degrees , would provide a duller blade generating more cavitation , drag and hemostasis . a longer , sharper taper would have substantially less tissue differentiation . the blade may have a continuous , sharp cutting edge as with conventional scalpels , or may have serrations or teeth as described below . referring now to fig5 a preferred shape of the serrations is shown enlarged . the serrations comprise parabolic - shaped recesses 44 separated by outwardly protruding teeth 46 . as seen in fig5 a , the teeth 46 also could have an alternate off - set or ` set ` where the blade includes hollow ground teeth . that is , the hollow ground , which forms the teeth , can extend into the blade from only one blade side and can alternate between the sides so as to give the blade edge 27 a wider kerf , as seen in fig5 a . this would cause additional drag , but also would allow better self - cleaning of the teeth 46 , especially on harder material which tends to clog teeth . with reference back to fig5 the teeth 46 desirably are spaced a certain distance apart to result in optimal cutting . advantageously , the teeth 46 are separated by a distance 46c of less than one longitudinal stroke of the blade 26 to ensure that the tips of at least two teeth 46 cross any one point in a single stroke . the spacing 46c of the teeth 46 is most preferably at most eight - tenths of the blade stroke so that every tissue bond is contacted by at least two teeth 46 during each stroke , while internal material stresses are minimized . it is understood that the effect of mechanical cutting increases as the number of teeth , which pass over a particular tissue bond during each stroke , increases . thus , the finer the teeth spacing for a given stroke length , the greater the effect of mechanical cutting . it has been determined that blades formed of an extremely hard material can support extremely fine teeth while withstanding the resultant internal material stresses produced by the ultrasonic vibrations at the cutting edge . for instance , materials with a rockwell ( c ) hardness of 55 or greater can support teeth having a spacing between about 1 to 10 microns , and materials with a rockwell ( c ) hardness of 65 or greater can support teeth having a spacing closer than 1 micron apart from one another . the material , however , should be selected such that the resultant internal material stresses produced by the acceleration and deceleration of the vibrating material stay below twenty - five percent ( 25 %) of the ultimate yield strength of the material . examples of suitable materials are diamond , sapphire , and composites formed by a metal base , such as , for example , aluminum alloy , high carbon stainless steel , or preferably titanium alloy , coated with a hard coating , such as , for example , a ceramic . preferred examples of suitable ceramics are tungsten carbide ( rc 74 ), titanium nitride ( rc 70 ), silicon - carbide ( rc 70 +), aluminum oxide ( rc 78 ), plasma - chrome oxide ( rc 74 ) ( i . e ., chrome oxide applied by a plasma ion application ) hard chrome plating ( rc 65 ), dense chrome plating ( rc 70 ), or electroless nickel plating after heat treatment ( rc 62 ). the advantageous shape of the teeth 46 of the blade 26 , shown in fig5 provides an enhanced feel of cut at all times . a straight - edged ultrasonic knife blade will slip through tissue with a substantially constant resistance due to the blade edge being everywhere parallel to the tissue . ultimately , the surgeon might apply more pressure than necessary , without realizing the depth of cut , and sever tissue not intended to be cut . the contour of the recesses 44 on the ultrasonic blade 26 of the present invention changes the angle of the portion of the blade edge which strikes the tissue . during light cuts , the surgeon notices little resistance as bond severing occurs primarily at the tip edges of the teeth 46 parallel to the plane of uncut tissue ahead of the cutting edge . to provide ample light cutting surfaces , the width 46b of the tips of the teeth 46 are preferably 30 - 60 % of the stroke amplitude , and most desirably the width 46b is 50 % of the stroke . slightly more pressure results in cutting at the sidewalls 45 of the recesses 44 , at least a portion of which is perpendicular to the plane of uncut tissue . the sidewalls 45 extend from the tip 46 of the teeth to the bottom 44 of the recess a sufficient distance to expose the perpendicular surfaces to the tissue . to ensure this exposure while retaining some strength for the extending teeth 46 , the depth 46a of the sidewalls 45 is 20 - 100 % of the blade stroke amplitude , and preferably the depth 46a is 80 % of the stroke . where a hollow - ground cutting edge 27 is used , it also is understood that if the depth 46a of the tooth 46 extends over the full radius of the curved surface 47 of the first taper 43 , cavitation effects will be increased and thermal effects will be reduced . an increase in the downward force causes more of the sidewalls 45 perpendicular to the tissue , between the teeth 46 to contact the tissue , resulting in a change of resistance due to the increased surface area contact at a high vector angle . thus , the surgeon experiences a greater resistance as the blade 26 is pressed harder into the tissue , and may adjust accordingly to prevent inadvertent injury to the patient . the surface texture of the blade 26 directly affects the amount of frictional and ultrasonic heat generation , in addition to the level of cavitation . highly polished surfaces tend to slide through the tissue with minimal friction and associated heat generation and sound transfer . the tapered surfaces 40 , 42 and the recessed region 36 are preferably polished to minimize thermal effects ( i . e ., thermal transfer ) to the tissue . concurrently , if dry cutting is preferred , the contact surfaces 38 may be finished slightly rougher to ensure heat will build up mostly at this region and increased hemostasis will occur . alternatively , the surfaces of the blade 26 may be roughened all over , a saline solution introduced at the operative site , and the blade oscillated at preferred rates to minimize thermal effects yet increase the amount of cavitation . such a situation is seen in brain surgery where a constant stream of water , or other coolant fluid , is applied to the incision area , and the majority of the cut is cavitation - assisted . referring again to fig2 and 3 , a transition section 48 alters the cross section of the blade 26 from the flat cutting section 34 to a generally cylindrical portion 50 comprising opposing wrench flats 52 . the transition section 48 amplifies the gain of the ultrasonic oscillations . a coupling member 54 adjacent to the cylindrical portion 50 mates with an opposite sex coupling member on the distal end of the handpiece 28 or an extender . due to the minimum time - constraints imposed by surgery , the coupling members are preferably rapid connect / disconnect types described below , with reference to fig9 - 16 , showing an alternative embodiment with an extender 55 . fig9 - 11 illustrate two junctions on either end of the extender 55 . fig1 shows a partial cross section of the coupling between the handpiece 28 and the extender 55 , and the extender 55 and the blade 26 . of course , it is understood that the coupling between the extender 55 and the preferred surgical blade 26 applies equally as well to a direct coupling between the blade 26 and the handpiece 28 . each junction comprises a generally cylindrical male component 56 and a tubular female component 58 comprising a generally cylindrical recess 60 adapted to receive the male component 56 . these components quickly connect by inserting the male component 56 into the female component 58 and rotating one component with respect to the other component , preferably through a relatively short rotational arc , and optimally about 90 °, plus or minus 10 °. only one junction will be referred to , as the junctions are identical . when joined , the junction produces a relatively high axial compression force , which is preferably uniformly distributed symmetrically about the contact surfaces between the two components to optimize the transfer of ultrasonic energy across the junction . non - uniform distribution of the axial compression force about the longitudinal axis of the junction tends to decrease the efficiency of the transfer of energy across the junction , and can cause unwanted transverse motion ( whipping ) and may lead to premature mechanical failure . although fig9 through 14 illustrate the male component 56 extending in a distal direction , it is understood that the relationship of the male and female components can be reversed . referring to fig9 - 12 , the male component 56 comprises at least two axially extending splines 62 spaced apart by at least two axially extending flats 64 . preferably , the male component 56 comprises two diametrically opposed splines 62 and two diametrically opposed flats 64 , alternatively positioned around the circumference of the component , as seen in fig1 . each spline 62 comprises a plurality of external threads 66 preferably configured in accordance with the american national standard for unified threads (&# 34 ; un &# 34 ;). it will be understood that other thread configurations , such as the american national standard acme screw threads (&# 34 ; acme &# 34 ;), can be used as well . it has been found preferable , however , to employ the un thread design instead of others , such as the acme thread design , primarily for manufacturing ease . advantageously , the thread pitch and the pitch diameter of the threads 66 and the length of the splines 62 are selected to produce high axial compression between the components without structural failure . it is also preferable to select a generally standard thread for manufacturing convenience . additionally , the threads 66 must engage to produce high axial compression with little rotation . preferably , circumferentially , 75 % of the threads 66 engage with rotation of no more than about 90 ° plus or minus 10 °. for example , in one preferred embodiment the splines 62 comprise a series of 4 - 56 uns - 2a threads 66 along a length of 0 . 215 inches , and in another embodiment , the splines 62 comprises a series of 5 - 48 unf - 2a threads 66 along a length of 0 . 250 inches . in general , the spline 62 preferably comprises about twelve interrupted threads 66 . in general , the junction has a minimum of 45 ° of total engagement between the spline threads 66 to produce the high axial compression without mechanical failure . preferably , the junction has an engagement between about 90 ° to about 179 °, and most preferably about 173 ° ( 48 % of 360 °= 172 . 8 °). thus , in a most preferred embodiment , the sum of the lengths of the threads 66 on the male component 56 measured in a circumferential direction preferably range from 90 ° to 179 °, and more preferably equal 173 °. the circumferential length of each spline thread 66 ( i . e ., the circumferential width of each spline ) depends upon the number of splines 62 employed . for example , in a most preferred embodiment having two splines 62 , the length of the thread 66 in a single spline along the circumferential direction ranges between 45 ° and 89 . 5 °, and preferably equals 86 . 5 °. the female component 58 likewise comprises at least two axially extending splines 68 and at least two axially extending flats 70 , disposed on the recess 60 circumference in a corresponding relationship with the flats 64 and splines 62 on the male component 56 , as best seen in fig9 and 16 . preferably , the female component 58 comprises two diametrically opposed splines 68 and two diametrically opposed flats 70 alternatively positioned around the circumference of the recess 60 , as best seen in fig1 . each spline 68 comprises a plurality of internal threads 72 configured to match and engage with the threads 66 on the male component 56 . as discussed above , the sum of the length of the threads 72 around the circumference of the recess 60 is preferably not less than about 90 ° and not greater than about 179 °, and most preferably equal 173 °. each spline thread length depends upon the number of splines 68 employed . for example , in a most preferred embodiment having two splines 68 , the threads 72 of each spline extend around the circumference of the recess 60 for at least approximately 45 °, but less than approximately 89 . 5 °, and preferably equal 86 . 5 °. the two splines 68 and two flats 70 alternately disposed on the interior circumference of the female component 58 recess 60 provide an axial key - way 74 for receiving the two opposing splines 62 on the male component 56 , as shown in fig1 . the male component 56 is inserted into the recess 60 of the female component 58 and rotated to interlock the corresponding splines 62 , 68 on the male and female components , as shown in fig1 . it is desired that minimum rotation of one component with resect to the other component will produce a junction which achieves a relatively high efficiency of energy transmission therethrough . in general , it has been found that a high compression across the junction symmetrically distributed about its longitudinal axis optimizes energy propagation . preferably , the thread design of the junction produces greater than about 100 pounds of axial compression force between the components with rotation of about 90 °± 10 %. more preferably , a compression in excess of about 200 pounds will be achieved . as a result of higher compression , the ultrasonic pressure wave propagates across the junction with minimal energy loss . it is preferred that the points of contact between the two joined surgical components be symmetric about the longitudinal axis of the male component 56 to uniformly distribute the compression force about the junction in the radial direction . as a result , the ultrasonic oscillation maintains its propagation along the longitudinal axis of the joined surgical components without deflection from that axis . if deflection occurs , the tool will tend to whip resulting in undesired heat build - up and loss of energy at the tool tip . in this regard , the female component 58 preferably additionally comprises an annular engagement surface 76 on the proximal end thereof which contacts a corresponding annular engagement surface 78 of the male component 56 . preferably , the engagement surface 76 of the female component 58 extends radially outwardly along a plane substantially perpendicular the axis of the internal recess 60 , and the engagement surface 78 of the male component 56 extends radially outward along a plane substantially perpendicular to the axis of the male component 56 . referring to fig1 , as the splines 62 , 68 , interlock , the two components draw together to force the engagement surfaces 76 , 78 , against each other , resulting in an axial compression force across the junction . preferably , the engagement surfaces 76 , 78 , are smoothly polished to produce a substantially liquid - tight seal between the components as the surfaces abut . in addition to optimizing energy propagation , a liquid - tight seal reduces cavitation erosion of the components at the junction and thereby extends the life of each component . in a preferred embodiment , the female component 58 additionally comprises an axially extending , generally cylindrical counterbore 80 at the distal end of the recess 60 for receiving a generally cylindrical shank barrel 82 on the proximal end of the male component 56 . the counterbore 80 and the shank barrel 82 are preferably centered with respect to the longitudinal axis of the male component 56 . preferably , the shank barrel 82 smoothly fits into the counterbore 80 to center the female component 58 with respect to the male component 56 . advantageously , the male component 56 further comprises an undercut region 84 positioned between the engagement surface 78 and the spline so that the spline threads 66 are fully formed ( i . e ., no run - out region ). as a result , the splines 62 , 68 can be reduced in overall length , as will be understood in the art . referring to fig1 , the female component 58 preferably additionally includes a generally cylindrical pilot recess 86 for receiving a corresponding generally cylindrical tip barrel 88 at the proximal end of the male component 56 . preferably , the diameters of the pilot recess 86 and the tip barrel 88 substantially coincide with the minor diameter of the threads 72 . advantageously , the pilot recess 86 and the tip barrel 88 are centered about the longitudinal axis of the male component 56 for optimizing the concentricity of the engagement surfaces , between the components to optimize the longitudinal transfer of ultrasonic energy through the junction . to facilitate rapid interconnection between the components , the female component 58 preferably additionally comprises an annular internal chamfer 90 and the male component 56 additionally comprises an annular tip chamfer 92 . when the male component 56 is inserted into the female component 58 , the chamfers 90 , 92 ease the insertion by funneling the components together . additionally , the edges of the leading spline threads 66 of the male component 56 preferably include a chamfer 94 to ease the engagement between the splines 62 , 68 of the male component 56 and female component 58 . referring to fig1 - 16 , it is preferred that the surgical components include alignment arrows 96 etched on the exterior surface of the components to aid in the connection process . by aligning the arrows 96 , the splines 62 of the male component 56 align with the key - way 74 of the female component 58 , as seen in fig1 and 15 . by rotating the components as shown in fig1 , the splines 62 , 68 of the two components interlock , as shown in fig1 . flat opposing surfaces 98 are provided on the exterior of all parts to receive a wrench to facilitate tightening and untightening of the junctions . those skilled in the art can manufacture the disclosed junction by processes known in the art . for example , the generally cylindrical male component 56 and the shank barrel 82 thereto can be cut into an end of the shank of a surgical component , such as the extender or the tool bit . the threads 66 can either be cold rolled onto the cylinder or preferably machine cut into the cylinder . the flats 64 can then be milled onto the component thereby interrupting the threads 66 . finally , the tip barrel 88 can be cut onto the distal end of the male component 56 such as by lathing operations well known in the art and the chamfers 92 , 94 , similarly added thereto . the recess 60 of the female component 58 can be made by drilling the pilot hole recess 86 into the end of a surgical component . the counterbore 80 then can be milled and a portion of the pilot hole 86 tapped with the appropriate internal threads 72 by processes known in the art . the flats 70 can be milled and broached into the recess 60 thereby interrupting the threads 72 on the recess wall . finally , the internal annular chamfer 90 can be drilled or milled to form a smooth transition from the counterbore 80 to the threaded recess 60 . referring again to the improved ultrasonic surgical knife system 24 of fig1 the control system 30 comprises an ultrasonic signal generator 100 which supplies an electric impulse to the handpiece 28 , the voltage of which can be varied at different frequencies and with different wave - shapes . the signal may , for example , be a pure sinusoidal wave or may be modulated with one or more other frequencies . alternatively , the signal may be a stepped or spiked pulse . in a preferred embodiment , the ultrasonic generator 100 transmits a signal of between 20 - 80 khz . more preferably , the signal is at about 60 khz . the signal generator 100 includes a liquid crystal or other display device 102 for convenient display of selected power or frequency mode . the signal generator 100 may , for example , transmit a constant amplitude signal at a constant frequency , or alternate one or both of these parameters . the cutting power level is normally selected as a percentage of maximum cutting power . although not illustrated in fig1 an audio output indicative of mode changes and present mode is preferably included which is responsive to the ultrasonic signal generator output 100 . the signal transmits through a multi - conductor shielded cable 32 , for safety and durability , to the handpiece 28 which imparts ultrasonic , generally longitudinal , movement to the surgical blade 26 . as will be described more fully later , high - efficiency piezo - ceramic washers 164 ( fig7 ) which generate the ultrasonic vibrations within the handpiece 28 ( fig7 ), allow a thin high - flex cable 32 to be used . the electronic signals are a lower than usual voltage not requiring a thick cable , which gives the surgeon added freedom to maneuver the handpiece 28 . a high quality autoclavable connector 106 couples the cable 32 to the signal generator 100 . referring to fig6 and 7 , the outer protective cover of the handpiece 28 generally comprises a nose cone 108 , a cylindrical casing 110 and an end cap 112 of durable stainless steel or other corrosion resistant material . advantageously , the protective cover is stainless steel and the sections are sealed hermetically , to protect the internal components from the corrosive fluids of surgery and temperatures in a steam autoclave . the handpiece 28 is preferably about 6 inches long and 1 / 2 inch in diameter . the distal end of the handpiece 28 is the end proximate the blade 26 , and the proximal end is the end proximate the cable 32 . an acoustic horn 114 transmits standing pressure waves from the piezo - ceramic washers 164 to the blade 26 . as used herein , the term &# 34 ; horn &# 34 ; means the component of the handpiece 28 that functions both as the acoustical concentrator , as well as the front end of the transducer . a central bolt 116 extends substantially the length of the handpiece 28 and provides a central coupling member rigidly joining the internal elements , as seen in cross section in fig8 . a heel slug 118 includes internal threads 120 for engagement with external threads 122 of the central bolt 116 . the horn 114 also includes internal threads 124 which couple with external threads 126 on the central bolt 116 . the piezo - ceramic washers 164 include a central bore 128 sized to fit over the external threads 122 of the central bolt 116 . the horn 114 and heel slug 118 compress the washers 164 therebetween via longitudinal movement along the central bolt threads 122 , 126 . the piezo - ceramic washers 164 , in combination with portions of both the horn 114 and heel slug 118 , comprise an electromechanical transducer , converting electrical energy to mechanical pressure waves . a rear annular bulkhead 130 is silver soldered to the rear of the central bolt 116 and supports the outer casing 110 at the proximal end of the handpiece 28 . the interface between the outer circumference of the bulkhead 130 and cylindrical casing 110 provides a hermetic seal and a solid ground connection . additionally , an o - ring 132 disposed between a front flange of the horn 114 and the nose cone 108 provides a fluid - tight interface . the piezo - ceramic washers 164 , and all other internal components shown in fig8 between the seals 130 , 132 , are thus enclosed within the cylindrical casing in a fluid - tight manner allowing the handpiece 28 to be immersed in a steam autoclave without harm . the horn 114 comprises generally three sections , a cross - sectionally enlarged section 134 , a transition section 136 and a narrow section 138 ( see fig7 ). the narrow section 138 at the distal portion of the horn 114 includes a female junction component 140 adapted to receive a male junction component ( not shown ) of a surgical blade 26 , or other surgical component . the mechanical energy which is produced by the piezo - ceramic washers 164 propagates along the horn 114 and amplifies at the transition section 136 . as is well known in the art , decreasing the cross section of a structure transmitting longitudinal pressure waves increases the stroke , i . e ., produces a positive gain in longitudinal oscillation . a stepped horn produces a gain which is approximately equivalent to the ratio of the larger area to the smaller area of the horn 114 , while a more gradual change in diameter produces a gain equivalent only to the ratio of the diameters . moreover , the location of the cross - sectional changes along the structure affects the degree of gain produced , as described below . thus , by adjusting the change in cross section of the horn 114 , the shape of the dimensional transition , and the location of the dimensional transition , a specific gain may be obtained to tailor the stroke of the blade 26 for optimum performance . preferably the gain achieved by the transition section 136 works in conjunction with a transition section of the blade 26 to produce an optimum longitudinal amplitude at the blade tip . the longitudinal amplitude of the blade 26 is preferably between 0 . 00025 and 0 . 004 inches peak - to - peak , and more preferably 0 . 0025 inches peak - to - peak at 60 khz , reducing the chance of material failure and controlling the energy for a fixed thermal footprint . the piezo - ceramic washers 164 remain in a stationary , compressed state between the horn 114 and the heel slug 118 and thus occupies a node of a standing wave created along the heel slug - washer - horn combination . at the nodes of vibration there is no motion but maximum stress . nodes are spaced exactly one half wavelength apart and thus from the piezo - ceramic washers 164 , nodes occur every half wavelength down the horn 114 ( e . g ., front transition 136 ). anti - nodes are points of absolute maximum amplitude , experience the largest longitudinal movement and the least stress , and are located 1 / 4 wavelength from each node . the closer the location of the cross - sectional change 136 to a node of vibration , the greater the gain realized , because the ultrasonic energy is stored as internal potential at these points , as opposed to kinetic energy at the anti - nodes . the elongated , cylindrical horn 114 preferably includes one step concentrator to tailor the gain to cause a preferred blade 26 to function optimally ; i . e ., to preferably stroke from 0 . 00025 to 0 . 004 inches , peak - to - peak , and more preferably at 0 . 0025 inches , peak - to - peak at 60 khz . the small stroke advantageously reduces internal stresses in the horn 114 and blade 26 and thus reduces the chance of material failure . the proximal end of the horn 114 defines an aperture leading to a central cylindrical cavity 142 sized to receive the distal end of the central bolt 116 . the cavity 142 includes internal threads 124 which mate with external threads 126 on the central bolt 116 . the cavity 142 extends axially in the distal direction , past the internal threads 124 , and ends at a chamfered portion 144 . the central bolt 116 includes opposing axial flats 119 for a wrench - assisted insertion into the cavity 142 . the majority of the enlarged section 134 comprises a solid cylinder to optimize ultrasonic energy propagation . the horn 114 is thus preferably constructed of a high strength material which efficiently propagates ultrasonic energy . more preferably , the horn 114 is constructed of titanium . the enlarged section 134 desirably has a length equal to approximately a half wavelength , with an anti - node positioned generally at the longitudinal center of the enlarged section 134 . in this manner , the front half of the enlarged section 134 forms part of the acoustical concentrator , while the back half forms the front end of the transducer . the distal portion of the horn 114 includes a female coupling portion 140 , as described above . the distal portion of the horn 114 additionally comprises a central lumen 146 extending proximally from the female coupling 140 preferably throughout the length of the narrow section 138 . the lumen 146 extends slightly past the transition section 136 . the lumen 146 assists in amplifying the ultrasonic energy propagated down the horn 114 . as described previously , pressure waves crossing a reduction in the cross - sectional area of a structure experience a gain . the lumen 146 defines a tubular section at the distal portion of the horn 114 , further reducing the cross - sectional area of the material of the narrow section 138 . in an alternative embodiment , as illustrated in fig8 a , the lumen 146a extends entirely through the handpiece 28a . for ease of understanding , like reference numerals with a prime mark (&# 39 ;) have been used to indicate like parts between the two embodiments . the lumen 46 &# 39 ; extends co - axially with the longitudinal axis of the horn 114 &# 39 ; and the central bolt 116 &# 39 ; and through the end cap 112 &# 39 ;. the end cap 112 &# 39 ; desirably is adapted to receive a tube coupling to couple the lumen 146 &# 39 ; with a fluid tubing . it also is contemplated that the blade / tool / sharps bit coupled to the ultrasonic handpiece 28 &# 39 ; likewise would include an internal lumen , which would communicate with the central lumen 146 &# 39 ; of the handpiece 28 &# 39 ; when coupled to the handpiece distal coupling 140 &# 39 ;. in this manner , the central lumen 146 &# 39 ; may be used either for irrigation or aspiration . with reference back to fig8 the overall length of the &# 34 ; horn &# 34 ; 114 for 60 khz is preferably less than about 2 . 5 inches , and more preferably the length of the horn 114 is 2 . 40 inches . the horn 114 is sized so that the front coupling junction 140 experiences a minimum of stress from being positioned close to an anti - node of vibration . the transition section 136 is desirably 10 : 1 or greater , and more preferably less than 0 . 75 of an inches from the farthest front portion of the horn 114 , and more desirably the transition section is 0 . 600 inches from the front of the horn 114 . the enlarged section 134 has a diameter of no more than 1 / 2 inch to fit comfortably in the hand of a surgeon , and more preferably the diameter of the enlarged section is 0 . 425 inch . advantageously , the inside diameter of lumen 146 in the narrow section 138 of the horn 114 is less than about 0 . 1 inches to provide a sufficient wall thickness of the frontal section to minimize stress failure . more preferably , the diameter of the lumen 146 is about 0 . 07 inches . the outer diameter of the narrow section 138 of the horn 114 is preferably no more than about 0 . 25 inches , and more preferably the outer diameter of the narrow section is 0 . 125 inches . advantageously , an exterior annular flange 150 at a position proximal to the transition section 136 provides a shoulder against which the o - ring 132 abuts . the nose cone 108 of the outer cover compresses the o - ring 132 rearward against the flange 150 in a semi - rigid manner , and in a fluid tight manner between the inside diameter of the tubing 110 and the outside diameter of the horn 114 . referring to the cross - sectional view of fig8 the length of the central bolt 116 is shown . the central bolt 116 comprises a solid , generally cylindrical metallic rod with a chamfer 152 at the distal end of a distal cylindrical portion 154 . the distal cylinder 154 fits in the distal cavity 142 of the horn 114 , as previously described . the distal chamfer 152 bottoms out at the internal chamfer 144 , providing a flush stop for the central bolt - horn interface , thus more efficiently transmitting ultrasonic energy . the distal thread region 126 separates the cylindrical portion 154 from a middle cylindrical region 156 . the threads 126 are preferably 0 . 2 inches from the front of the central bolt 116 , and the proximal section of threads 122 is located 1 . 4 inches further rearward . preferably , the threads 126 are 10 - 56uns - 2a type threads , and configure to meet with similar internal threads 124 of the horn 114 . the middle cylindrical portion 156 extends through the central bore 128 of the piezo - ceramic washers 164 . the washers 164 slide along the middle portion 156 to abut the horn 114 adjacent the distal threads 126 of the central bolt 116 . the distal axial face of the washers 164 and proximal axial face of the horn 114 lie flush against a thin annular spacer 170 therebetween to optimize transmission of ultrasonic vibrational energy . the proximal thread region 122 separates the middle region 156 from a cylindrical heel slug receiving portion 158 . the bolt 116 terminates in a reduced diameter isolation region 160 and a rear bulk head support shaft 162 . the rear thread 122 region is adapted to receive the heel slug 118 . as stated previously , the heel slug 118 threads onto the central bolt 116 , compressing the piezo - ceramic washers 164 against the horn 114 . the rear - most portion of the heel slug 118 terminates at the transition of the central bolt 116 to the isolation region 160 . the large change in diameter between the heel slug 118 and the isolation region 160 causes the isolation region 160 to tend to vibrate at its own natural frequency , interfering with sound propagation at the fundamental frequency in this direction . the ratio between the cross - sectional areas of the heel slug 118 to the isolation region 160 desirably is 10 : 1 or greater , and more preferably is 25 : 1 or greater . this abrupt cross - sectional area change in the transducer configuration causes less than 5 percent loading on the overall handpiece 28 to essentially isolate the handpiece 28 from the produced vibrations . in this manner , little ultrasonic energy is propagated rearward . in addition , the additional 1 / 4 wavelength length of the bulk head support shaft 162 on one side , and the isolation region 160 of a length less than 1 / 4 wavelength -- preferably about 1 / 16 to about 1 / 8 wavelength -- on the other side forces the bulkhead 130 to be an artificial or virtual node ( a node and an anti - node separated by less than λ / 4 ). the 1 / 4 wavelength length of the bulk head support shaft 162 thus functions as a terminal resonator which is desirably tuned to reflect vibrations in phase with the vibrations propagating from the distal side of the piezo - ceramic washers 164 down the horn 114 and the attachments thereto ( e . g ., extender ( s ), tool / blade / sharp bit , etc .). this reinforces the stability of the bulkhead 130 location and minimizes any loading of the handpiece when the bulk head 130 is silver soldered to the central bolt 116 and the inside diameter of the tube 110 . the combination of the cross - sectional area ratio between the heel slug 118 and the isolation region 160 of the central bolt 116 , the location of this area reduction at an anti - node , the one - quarter wavelength length of the bulk head support shaft 162 , and the overall configuration provides good acoustical isolation ( i . e ., 5 percent or less transmission ), good electrical conductivity between the components of the handpiece for a good ground path , and good mechanical stability ( i . e ., torque resistance ) to prevent the horn 114 from rotating when coupling a tool / blade / sharp to the distal coupling end 140 . in addition , where the central lumen 146 &# 39 ; extends through the entire handpiece 28 &# 39 ; ( see fig8 a ), this integral construction provides a uniform , continuous , low - resistance flow path between the proximal and distal ends of the handpiece 28 &# 39 ;. the material of the thin annular washers 164 is a piezo - ceramic compound of lead - titanate and zirconium titanate . advantageously , two to eight washers 164 may be utilized , depending on the strength of vibration desired , and preferably there are two washers 164 . these washers 164 include central bores 128 to fit over the middle cylindrical region 156 of the central bolt 116 . the central bore 128 passes over the rear threads 122 , and an insulating material 168 is wrapped around the central region 156 to fill the annular void formed and hold the washers centered on the bolt 116 . two very thin annular spacers 170 separate the piezo - ceramic washers 164 from the horn 114 and heel slug 118 , and distribute the compressive forces evenly . a layer of electrically insulating material 166 covers the washers 164 and isolates them from the outer casing 110 of the handpiece 28 . an air gap 172 between the insulating layer 166 and the casing 110 provides effective thermal and acoustical isolation from the outer casing . preferably , the air gap 172 is approximately 0 . 017 inches , which has been found to reduce the radiation of internal heat to the outer casing 110 . a &# 34 ; hot &# 34 ; electrode 174a and a ground electrode 174b connect to the appropriate piezo - ceramic washer 164 to effectuate mechanical vibrations . the electrodes 174 extend proximally from the piezo - ceramic washers 164 within the air gap 172 . the &# 34 ; hot &# 34 ; electrode 174a passes through a small passage 176 in the bulkhead 130 and from there to the rear end cap 112 and a &# 34 ; hot &# 34 ; circuit of the connector 106 of the cable 32 . the ground electrode 174b connects directly to the bulkhead 130 which is in electrical contact with the ground circuit of the connector 106 . as is well known in the art , piezo - ceramic materials produce mechanical vibrations upon excitation by an applied voltage . this mechanical vibration is caused by changes in the internal structure when under the influence of the external voltage . the piezo - ceramic washers 164 are held under compression between the horn 114 and the heel slug 118 . preferably , the compression of the piezo - ceramic washers 164 is between 500 and 5000 psi , and most preferably about 1500 psi . aligning the washers so that the positive side of one abuts the positive side of another causes the washers to oppose each other &# 39 ; s motion , and in effect double their amplitude vibrations . such piezo - ceramic washers 164 held in compression are restricted from thickening ; their internal stresses are transmitted to the surrounding compressive members in the form of pressure waves . the preferred piezo - ceramic configuration is a &# 34 ; langevin sandwich &# 34 ; design . as the waves propagate along the horn 114 and heel slug 118 , the potential strain energy converts to kinetic energy and back , due to the wave - like nature of the signal . the heel slug 118 and adjacent isolation region of the central bolt 116 tend to reflect the vibratory motion with some losses ( damping ) while the excellent energy transmittal properties of the titanium horn 114 propagates the vibrations directly to the blade 26 with minimal losses . the washers are thus aligned and compressed between the heel slug 118 and the transducer / horn 114 . the compression of the piezo - ceramic washers 164 results in standing pressure waves propagated down the horn 114 and reflected back . as stated previously , the piezo - ceramic washers 164 preferably occupy a node of vibration and other nodes appear approximately one - half wavelength later ( if the material or cross section does not change ) and every half wavelength subsequently . anti - nodes are located approximately 1 / 4 wavelength between the nodes and experience the largest longitudinal movement and the least stress . at 60 kilohertz , each 1 / 2 wavelength equals approximately 1 . 6 inches in the preferred titanium horn 114 . the horn 114 is machined so that the transition region 136 desirably occupies a node . in addition , the coupling region 140 at the front portion of the horn 114 is preferably placed close to an anti - node to reduce the stress of the coupling . thus , locations of the transition region 136 and the front coupling region 140 are multiples or fractions of the preferred 1 / 2 wavelength of 1 . 6 inches . the heel slug 118 is preferably fabricated from tool steel or stainless steel . a central bore 178 extends through the heel slug 118 and includes internal threads 120 at the rear ( proximal ) end . the heel slug 118 also comprises two opposing wrench flats 180 at the rear end . the parameters of the blade 26 may be altered , or the ultrasonic signal may be varied , to customize the type and character of incision desired . as stated previously , higher frequency surgical knives tend to propagate energy shorter distances into surrounding tissue and thus propagate thermal effects to less depth . at times though , some thermal effect on the tissue is desirable , especially when dry cutting . modulating a high frequency signal with a substantially lower carrier frequency allows the surgeon to nominally retain the advantageous features provided at high frequencies ( hemostasis ) while periodically applying a lower frequency to effectuate some increased degree of cavitation . at lower frequencies there is more drag , and thus more feel and tissue differentiation . adjusting the modulating frequency to decrease the periods of high frequency results in more feel , and thus the surgeon may selectably alter the response of the surgical blade 26 to different types of tissue . in a preferred embodiment , the surgical blade 26 of the present invention is vibrated at 60 khz with a modulating frequency of between 10 and 10 , 000 hz , and a preferred frequency of 600 hz . this is a concession to ergonomics only . loud , audible frequencies above 600 hz are more irritating ; otherwise , the modulation frequencies of choice to maximize cavitation would be about 5 to about 25 khz , with about 10 to about 15 khz the more preferred and about 10 khz the most preferred . referring now to fig1 and 18 , a split chuck 182 connects with an extender which couples with the handpiece 28 in an alternate form of the present invention . the split chuck 182 is shown in greater detail in fig1 . the split chuck 182 includes a forward slot 184 which receives a flat surgical cutting blade 186 . the blade 186 is placed within the slot 184 and a collet 188 threads over the chuck 182 to tighten the blade within the chuck . chuck 182 is provided with opposing wrench flats 190 to tighten the chuck in the handpiece 28 , or an extender , with a wrench . advantageously , the extenders allow the handpiece of the present invention to be remain external to the body while the blade extends within a catheter , trocar sheath , or endoscope lumen for endoscopically - assisted surgery . the extenders possess excellent sonic transmission properties with minimal losses at the interfaces . additionally , the rapid connect / disconnect coupling feature allows rapid changing of extenders , blades and chucks . preferably , the present invention may be used endoscopically with a 4 millimeter catheter opening . preferably , extenders allow surgery at a depth of as much as 24 inches from the handpiece 28 . referring to fig1 , there is disclosed a blade carrier 195 in accordance with a further aspect of the present invention . blade carrier 195 facilitates handling of the ultrasonic surgical blade in a sterile environment prior to installation on an ultrasonic handpiece . in addition , the use of the blade carrier 195 minimizes the risk of inadvertent blade sticks during handling and installation , removal , and disposal of the blade . blade carrier 195 generally comprises a blade housing having a blade connector end 196 , and a blade tip end 198 . the overall length of the blade carrier 195 is preferably about 2 . 5 inches . blade cavity 200 is disposed therebetween , for receiving the sharp end of the blade . the connection end of the blade , which may be threaded or provided with other quick connection / disconnection means previously disclosed , projects from the blade cavity 200 axially through the open channel 204 and out the open end 210 . the open channel 204 is provided with a pair of opposing surfaces 206 and 208 for frictionally engaging the wrench flats on the connector end of a blade , as has been previously described . referring to fig2 , opposing surfaces 206 and 208 can be more clearly seen . in a preferred embodiment , projections 209 and 211 are additionally provided for retaining the connection end of the blade within the open channel 204 . the blade cavity 200 is a shallow flat or rounded bottomed recess , having a length dimension 201 of about 1 . 5 inches and a width dimension 202 of about 0 . 50 sufficient to accommodate a variety of blade configurations . in general , blades contemplated to be utilized with the ultrasonic knife in accordance with the present invention have a cutting edge length within the range of from about 0 . 5 inches to about 1 . 5 inches . in addition , the width along the plane of the blade varies within the range of from about 0 . 030 inches to about 0 . 40 inches for most applications . specialty blades , for unique applications , may vary considerably from the foregoing ranges . the blade carrier 195 is provided at its blade tip end 198 with a knob 212 . knob 212 comprises a generally cylindrical body , preferably having a diameter of about 0 . 50 inches , and length of about 0 . 5 inches , having friction enhancing structures such as knurling on the radially exterior wall thereof . the axis of knob 212 is aligned with the axis extending through the open channel 204 . in this manner , the clinician can spin the knob 212 between two fingers to threadably engage the connector on the knife with the corresponding connector on the ultrasonic handpiece or extender as discussed below . the blade carrier 195 is preferably also provided with a pair of opposing wings 214 and 216 to provide leverage for rotating the blade carrier to tighten the connection between the blade and the ultrasonic knife , extender , or handpiece . preferably , the overall width of the carrier through the wings 214 , 216 is about 1 . 2 inches . as has been previously discussed , the typical connection between the knife tip and the handpiece is a rotatably engageable connection . for example , with the quick connect and disconnect embodiment previously disclosed , the blade is inserted onto the handpiece or the connector by an axial advancement and then the blade is tightened by rotating the blade through an angle of approximately 90 °. in an alternate embodiment , the blade is simply threaded onto the handpiece or connector by rotating through a series of complete revolutions . in either embodiment , the blade must be appropriately rotationally tightened into the handpiece or extender . for this purpose , the opposing surfaces 206 and 208 and a hinge region 207 therebetween are preferably molded from a material having a suitable resilience that the rotation of the blade carrier 195 will rotate the blade contained therein until the blade is suitably tightened against the handpiece or extender . further rotation of the blade carrier 195 will cause the opposing surfaces 206 and 208 to spread slightly , permitting relative rotation between the blade carrier 195 and the blade contained therein . the clinician simply rotates the blade carrier 195 until the assembly &# 34 ; snaps &# 34 ; or starts to cam over . in this manner , a predetermined predictable and repeatable amount of torque within the range of from about 0 . 50 to from about 80 . 0 inch - lbs ., preferably about 3 . 0 inch - lbs ., can be applied during installation of the blade . wings 214 and 216 provide both a friction surface and leverage for the clinician to use to rotate the blade carrier 195 during installation . following sufficient tightening of the blade , the blade carrier 195 is simply pulled laterally away from the tip of the blade and discarded , or saved , to be reinstalled at the end of surgery and then discarded with &# 34 ; sharps .&# 34 ; the blade carrier 195 may be constructed in any of a variety of ways which will be well known to one of skill in the art . for example , the entire blade carrier may be integrally molded such as by injection molding , thermo forming or vacuum forming of a pre - formed sheet of plastic . alternatively , the blade carrier 195 can be fabricated from premolded component parts , such as by premolding the blade connection end 196 and the knob 212 . the main body of the blade carrier 195 is preferably stamped or molded from a sheet of plastic , and may be thereafter secured to the blade connector end 196 and knob 212 using thermal bonding , solvent bonding , ultrasonic welding or other techniques known in the art . alternatively , some or all of the blade carrier 195 can be formed from an appropriate metal sheet , and preferably thereafter provided with an appropriate plastic coating . in general , the construction of the blade carrier 195 is of appropriate materials that will permit sterilization of the assembly of the blade carrier 195 with a blade therein . the blade carrier 195 and blade are thereafter introduced into a sealed packet or pouch for sterilization and shipment . problems associated with ultrasonic surgery can be generally classed in two categories . the first category would be the effect on the living tissue on either side of the cut . excess heat generation , tearing of tissue or inadvertent cutting of nearby anatomical structures are all problematic to ultrasonic surgery . the second category of problems is a relative lack of operator comfort , flexibility and feedback . in ultrasonic surgery , the knife blade may oscillate at anywhere within the range of from about 1 khz to about 100 khz . typically , however , frequencies of lower than about 23 khz are not used because they are within the audio range . in addition , frequencies in excess of about 50 or 60 khz produce an increased amount of localized heating along the tissue contacting sides of the blades . for relatively low frequencies , e . g ., below about 20 or 30 khz , high carbon steel or stainless steel is an appropriate construction material for the ultrasonic knife blades of the present invention . however , frequencies in excess of about 30 khz , which are considered relatively high , are preferably used in conjunction with ultrasonic knife blades made from or coated with titanium , aluminum , or other metals or alloys which will transmit ultrasonic energy efficiently , with less internal heating . approximately 50 % of the heat is produced from sound absorption in the surrounding tissue , 25 % produced from internal frictional heating of the blade itself , and 25 % produced by friction of the blade and tissue . at times , heat is preferred if a hemostatic nature of cut is desired . hemostasis is the coagulation or formation of white gelatinous substance at the sides of the cut , and is commonly referred to as &# 34 ; bloodless surgery .&# 34 ; at temperatures above 65 ° c . ( 149 ° f . ), proteins in human tissue are denatured , producing coagulation . although in some instances hemostasis is desirable , the increased temperatures involved in ultrasonic surgery potentially increase the likelihood of denaturing protein in tissue and can produce localized thermal damage , or necrosis , to the tissue surrounding the incision . as mentioned above , sound absorption into the surrounding tissue comprises the majority of heat generation in ultrasonic surgery . ultrasonic surgical instruments propagate pressure waves down the blade and into the surrounding tissue . at the interface of the blade material and the tissue , there is an impedance mismatch , causing the sound waves to dampen or &# 34 ; deaden &# 34 ; as they attempt to propagate further into the tissue . the energy absorbed by the damping characteristics of the tissue is converted to heat . preferably , the ultrasonic energy does not propagate far into the tissue , to limit the effect the heat produces . it is well known that higher frequency , shorter wavelength signals dissipate faster and in shorter distances in elastic material , such as biological tissue , and therefore would appear to be favorable to limit the depth of thermal effects . a smaller percentage of the total heat produced in an ultrasonic surgical procedure occurs from internal frictional heating of the knife blade . in general , the construction material of the surgical blade determines the level of internal friction , and potentially damaging heat . stainless steel , for example , is a relatively inefficient conductor of acoustic energy , and a lot of internal friction results . stainless steel used at frequencies above about 20 khz can get very hot . the titanium used for the present surgical blade 26 on the other hand is an excellent conductor of acoustic energy and may be used at the highest frequency contemplated ( 60 khz ) with a minimum heat buildup , especially if caused to start and stop vibrating intermittently . however , as stated previously , some heating may be necessary if bloodless surgery is desired . sharp surgical blades oscillating at ultrasonic frequencies can tend to fall through living tissue , much like a hot knife through butter . conversely , a duller knife , or one which has low or no ultrasonic assistance , requires a greater amount of force , and more subsequent tearing of the tissue occurs . such slower cutting , which may result in more scarring , may be desirable when performing surgery proximate vital organs so that the surgeon can feel the blade advancing through the tissue and more carefully continue . ultimately , ultrasonic surgery results in the breaking of living tissue bonds which are of varying strengths . the present invention addresses this issue and provides the surgeon with multitudes of configurations of blades , depending on the type of cut desired . the second category of problems associated with ultrasonic surgical tools are those relating to the lack of operator control of , and poor ergonomics of , the instruments . first , there has been a lack of understanding of the tactile feedback necessary to carefully resect different types of living tissues with one particular knife . as discussed above , a very sharp knife might be desirable , for instance in cosmetic surgery , but provide the surgeon with little or no feedback of the type of tissue the knife is cutting through . conversely , a knife with lots of drag may provide feedback , but may have a substantial reduction in the quality of cut desired . additionally , the amount of feedback desired is a subjective determination by the individual surgeon . a more experienced surgeon would tend to require less feedback than a novice . the amount of heat generated is another critical control parameter , previously addressed by simply altering the thermal footprint of the blade . another phenomenon associated with ultrasonic surgery is the formation of cavitation bubbles in the region proximate the surgical blade . control of the amount of cutting from mechanical shearing of the tissue bonds , as opposed to that from cavitation - assisted cutting , has not previously been addressed . cavitation occurs when the local pressure in a fluid decreases below the vapor pressure of that fluid . local voids or vacuum pockets , in effect , are created which then tend to implode violently upon an increase in pressure . objects moving rapidly through a fluid can induce such cavitation in their wakes by abrupt changes in fluidic pressure , as is known in the art of fluid dynamics . ultrasonically oscillating surgical blades have a tendency to cavitate in the bodily fluids surrounding an incision . in addition , normal saline or other fluids can be supplied to a surgical site to enhance normal cavitation . the saline or irrigation solution desirably includes a biocompatible element to introduce micro - weaknesses in the solution . carbonate salts , dissolved gases , and other compounds or particulates can be suspended or dispersed in the solution as the biocompatible element . the presence of such elements lowers the energy threshold requirements for effective cavitation . for instance , an effective power cavitation threshold can be reached at 40 khz with a 0 . 0012 - inch peak - to - peak stroke length in a well - filtered , degassed irrigation solution . with an enhanced salt solution ( i . e ., an increased amount of biocompatible carbonate salts ), the power threshold can be lowered to produce a 0 . 0006 - inch peak - to - peak stroke length while obtaining effective cavitation . at lower frequencies , e . g ., below about 20 to 30 khz , ultrasonic knives tend to create a cavitation emulsification layer which nominally provides better lubrication for the knife blade , and tends to minimize the effects of heat transfer to the surrounding tissue . the amount of cavitation plays a major role in the characteristics of the final cut . the implosion of cavitation &# 34 ; bubbles &# 34 ; can be detrimental to the micro - surface of the surgical instrument , but also can assist the cutting action by breaking tissue bonds at the same time . the physics of the formation of cavitation bubbles is such that the temperature at their surface can reach 5000 ° f . this intense but highly localized energy is converted to the kinetic energy of a shock wave upon implosion . the result is that the knife tends to &# 34 ; blow through &# 34 ; the tissue and the energy which would have been converted to thermal transfer to the surrounding tissue is used for cutting . in some instances cavitation primarily , in conjunction with some hemostatic action , is a preferred cutting method . the present invention addresses the aforementioned problems associated with ultrasonic surgery in terms of varying the characteristics of the incision and providing the surgeon with proper feedback and flexibility of use . the surgeon has a wide range of blade configurations and operating modes to best perform a particular procedure to his or her preference . the tactile feedback and cutting options available with the present ultrasonic surgical blade are a major improvement over prior instruments . the amount of heat generated and propagated into the surrounding tissue is controlled by the shape of the preferred blade 26 . the area of the contact surfaces 38 can be widened to increase the heat generation from sonic and conductive energy transfer . this is desirable in regions containing numerous blood vessels to induce hemostasis . similarly , the angle of the tapers 40 , 42 affects the magnitude of thermal footprint of the blade 26 . a large portion of the cross - section of the blade 26 in contact with the tissue being cut is removed by the formation of the relief 36 . these parameters can be cohesively managed to provide a wide range of incision characteristics . for example , cosmetic surgery requires the sharpest blade with minimal thermal damage to minimize scarring . alternatively , a sharp blade with more heat generation may require a similar blade tip with more contact surface and less relief in the central portion . the hollow - ground blade edge also brings these two effects closer together to provide a very sharp knife with substantial heat generation directly behind the cutting edge for increased hemostatic tissue activity . the various shapes of the current blade 26 contemplate an infinite number of functional combinations . another factor in heat generation is the surface texture of the blade 26 surfaces . smoother surfaces result in less frictional resistance than rougher ones . roughening the surface texture of the contact surfaces 38 , while highly polishing the tapers 40 , 42 and relief portions 36 , results in some increase in heat generation , which can be customized for the type of tissue involved . surface textures can be modified by either polishing an existing surface or roughening the existing surface of the blade . minimal surface friction will be incurred in a blade having a highly polished surface such as an rms of 1 or 2 . rms , or root - mean - square , is a proportionate term generally referring to the statistical average of the sizes of irregularities . practically , however , polishes of this degree are difficult to produce on the construction material utilized for surgical blades . relatively rough areas of the surgical blades disclosed herein are contemplated to have an rms of about 63 . this level of roughening can be accomplished by processing the knife blade with glass beading , chemical etching , or other techniques which will be known to one of skill in the art . preferably , a random sized distribution of bumps or pockets within the range of from about 20 to about 400 micron are utilized when operating in an ultrasonic frequency range of about 20 khz . the bumps or pockets are preferably rounded or hemispherical in shape , to improve longevity under ultrasonic vibration conditions , and to minimize fragmentation and leaving parent material behind . it is understood that the size of the irregularities or micro cavities in the cutting edge surface can be decreased in size if the knife is intended for operation in a higher frequency range . for instance , if operated in an ultrasonic frequency range of about 40 khz , the irregularities can have sizes ranging between about 10 to about 200 microns , and if operated in a frequency range of about 60 khz , the irregularities can have sizes ranging between about 5 to about 100 microns . at higher frequencies ( e . g ., 100 khz ), the irregularities sizes can be smaller ( e . g ., 1 . 0 micron or less ). a further parameter influencing the amount of thermal generation is the frequency and mode of oscillations . the control system 30 of the present invention allows for complete flexibility for the surgeon to alter the oscillation character . as is known , a higher frequency surgical blade tends to transfer less thermal energy to a greater depth via sound propagation to the surrounding tissue but has higher internal heat of blade and at the interface of blade and tissue . the control system 30 provides a means for modulating such an advantageous frequency with lower frequencies to provide some drag , or tactile feedback , to the surgeon , and increase effective cavitation . other combinations of frequencies and waveforms can be generated by the control system 30 to tailor the oscillations of the blade 26 to the particular surgical environment . for instance , as discussed above , both the amplitude and the frequency of a signal driving the blade can be modulated to produce effective heating and cavitation cutting . by way of an example , the amplitude of a base signal ( e . g ., 60 khz ) can be modulated between about 5 % and about 100 % of its peak amplitude . that is , the control system 30 steps the amplitude of the base signal between about 5 % and about 100 % of the peak amplitude . ( as noted above , the peak amplitude of the driving signal is selected to produce the desired stroke length of the blade ). the period between modulations desirably can be as low as 600 hz and as high as 15 khz . during periods of peak amplitude , the base signal also can be modulated with a high frequency ( e . g ., 90 khz ). as discussed above , the modulation of the base signal with a high frequency provides excellent heating effects at the knife / tissue interface for tissue hemostatic activity , while amplitude modulation of the base signal at a low frequency increases cavitation formation to assist cutting . the present invention also identifies and presents solutions to the problems of feedback and individual surgeon needs . the advantageous shape of the serrations of the present blade 26 transfer resistance forces more efficiently to the hand of the surgeon . providing surfaces perpendicularly vectored to the tissue means that more resistance is encountered from an increase in the pressure of cut . reducing the stroke of the blade and spacing the teeth 46 of the blade 26 so that at least two teeth 46 encounter a specific tissue bond on each stroke reduces the internal stresses on the knife as well as the magnitude of vibrations of the handpiece , while ensuring a clean and effective cut . another benefit of the ultrasonic surgical system 24 of the present invention is the ability to manage the amount of cavitation generated . cavitation minimizes thermal energy penetration into the surrounding tissue by converting the transient shock wave energy into a cutting action . the dynamic feedback associated with cavitation - assisted cutting provides enhanced tissue differentiation , as the stronger , more elastic , bonds holding such anatomical structures as blood vessels together require more energy to break than does the surrounding tissue . the feedback from cavitation cutting , in effect , increases the surgeon &# 39 ; s feel for changes in drag felt when cutting from weak to tough tissue , as opposed to the coarse changes in feedback from simply mechanically shearing the same tissue layers . control of the various parameters of the present invention allows the surgeon to select the amount of cavitation produced . the primary factor for changing the amount of cavitation at a fixed frequency and a uniform saline solution is the surface texture of the blade 26 surfaces . smoother surfaces result in less frictional resistance than rougher ones and thus less disturbance of the fluid boundary layer next to the blade . roughening the surface texture of a blade results in wakes and the subsequent formation of additional cavitation bubbles . in general , the larger the surface irregularity , the larger and more energetic bubbles are formed . the discussion of surface roughness of the blade 26 above in terms of preferred frictional heating applies to cavitation as well . relatively rough areas of the surgical blades disclosed herein to induce a substantial amount of cavitation are contemplated to have an rms of about 63 . cavitation can also be increased by increasing the angle and width of the blade cross - section which contacts the tissue . the shape and surface texture of the teeth of the present blade can be altered to increase or decrease cavitation or , in effect , manage the percentage of cutting due to cavitation . for instance , as mentioned above , a blade edge having a hollow - ground first taper and a tooth depth 46a which extends over the full radius of the curved surface of the first taper will increase the amount of cavitation . cavitation is highly dependent on the frequency of oscillation . lower frequencies , in general , produce more cavitation as slower moving blades tend to form larger bubbles ; there is approximately nine times more cavitation energy at 20 khz than at 60 khz for the same stroke ( peak to peak motion ). the present invention advantageously can be configured to increase the amount of cavitation at higher frequencies . in addition to altering the shape and texture of the blade , the blade 26 oscillation may be started and stopped with gated pulses to induce more cavitation . a blade operating at 60 khz to take advantage of the reduced thermal penetration , for example , may be gated to cause a greater number of larger cavitation bubbles to form during the slow - down and start - up periods without increasing the thermal effect on the surrounding tissue . the depth of thermal penetration is desirably limited to 1 mm into the sides and bottom of an incision . advantageously , the gated pulses would be applied directly out of phase from the original frequency to rapidly dampen out the natural vibration of the oscillating blade 26 and horn 114 . the gated pulse would preferably only reduce the vibrational amplitude to s - 5 - 10 % of the original and thus leave the blade and horn &# 34 ; singing &# 34 ;. the start up pulse would then be applied directly in synchronous phase with the small residual vibrations , to more quickly bring the blade 26 and horn 114 back to the original amplitude . another primary advantage with the surgical knife of the present invention is seen in its ability to cut through a wide range of materials with a maximum of control . coordinating the blade 26 configuration , ultrasonic signal shape and surgical technique permit an infinite number of applications . for example , in the area of tissue resection , straight cutting or dry cutting with hemostasis , or cavitation - assisted cold - cutting are all within the realm of uses for the present invention . similarly , other more durable materials may be cut with the present blade 26 . osseous matter can be sawed easily and with minimal necrosis . plastics and cements , such as pmma used in affixing prosthetic devices within bodycavities , are also rapidly cut through with the proper toothed blade 26 and at the proper frequency . another possible use for the present invention is for delaminating hi - tech composites , the vibrations serving to break the chemical bonds of the laminates . a further configuration possible with the blade 26 of the present invention is machining more than one shaped edge around the blade . this time - saving feature would provide a surgeon with essentially two or more tools in one . normally , a surgical incision passes through many different types of tissue , requiring different techniques or a new blade altogether . the time spent switching a blade can be extremely costly to the patient . the present surgical blade 26 may have one side shaped and finished for rapid , sharp cutting through outer layers of tissue . the other edge of the blade may have a rougher wider shape to induce more cavitation and drag , for &# 34 ; teasing &# 34 ; the blade through tissue close to vital organs . other possibilities include edges preferred for cold - cutting ( more cavitation ), cauterizing ( localized heating ) or bone cutting ( minimum heating ). finally , the variations of blade and oscillation character provide the knowledgeable surgeon with a highly advanced and flexible surgical tool . the numerous combinations of the aforementioned surgical knife parameters give the surgeon ultimate freedom in choosing the preferred embodiment . the present invention has been described in terms of certain preferred embodiments . however , additional embodiments and variations will become apparent to one of skill in the art in view of the disclosure contained herein . such variations are intended to be within the scope of the present invention . accordingly , the scope of the present invention is not limited by the specific embodiments disclosed herein , but is to be defined by reference to the appended claims .

disclosed is an improved ultrasonic knife of a type for incising various types of material . the knife has a reduced thermal footprint to tailor thermally induced tissue effects . tooth configuration on the knife cooperates with the stroke of the ultrasonic drive to produce efficient cutting , as well as tactile feedback to the surgeon with respect to the rate of cutting , and changes in tissue density . ultrasonic knife tip extenders are also disclosed for advancing the ultrasonic knife tip through the working channel of an endoscope or trocar sheath . methods utilizing the foregoing apparatus are also disclosed .

fig1 illustrates a common set - up for assisting a patient 1 to breathe with a negative pressure ventilator 3 . in this set - up , the patient 1 is lying supine on the bed 5 and the shell 7 is positioned above his chest . the shell 7 is held in place by velcro or other removable strap arrangements 9 with the perimeter edge at 11 of the shell 7 substantially conforming to the patient &# 39 ; s body to form a seal . the main portion 13 of the shell 7 is slightly spaced from the patient &# 39 ; s chest to form a chamber 15 . in operation , the negative pressure ventilator 3 periodically draws orsucks air out of the chamber 15 through the connecting tube 17 . this causesthe patient &# 39 ; s chest to be expanded upwardly into the chamber under the influence of the ambient air . that is , the ambient air which is at a relatively higher pressure seeks the lower pressure in the chamber 15 . as a consequence , the ambient air enters into and expands the patient &# 39 ; s chestinto the chamber 15 filling his lungs with air . upon release of the negative pressure by the ventilator 3 , the patient &# 39 ; s chest then essentially falls under its own weight to expel the breath from his lungs . this fundamental manner of operation is old and well known . the negative pressure ventilator system of the present invention is shown in cross section in fig2 . as shown , the connecting tube 17 which is the primary flow channel to the patient is attached in a simple , overlapping manner to the passageway 19 . the passageway 19 is part of the valving housing 21 and as illustrated is located at the bottom of the valve housing 21 of the overall valving arrangement 23 . in the position of fig2 air is being drawn through the connecting tube or primary flow channel 17 from the shell 7 ( fig1 ) on the patient &# 39 ; s chest under the influence ofthe turbine 31 of the air blower 33 . more specifically , the turbine 31 of the air blower 33 is being rotated by the motor 35 to move air through themain body 37 of blower 33 . the inlet 29 to the main body 37 of the blower 33 is thus at a negative pressure less than ambient or atmospheric pressure . the outlet 39 is then at a positive pressure greater than ambient or atmospheric pressure . ( such negative pressure relative to ambient pressure is indicated in fig2 and the other drawings by hollow arrows and such positive pressure relative to ambient pressure is indicated in fig2 and the other drawings by solid arrows .) the three dimensional nature of the valving or control arrangement 23 between the primary flow channel 17 and the blower 33 is somewhat difficult to describe and illustrate simply by reference to only one or two figures . however , suffice it to say at this introductory point in the disclosure that the air flow through the apparatus in the position of fig . 2 is as follows . using the shell 7 of fig1 as a beginning reference point , the air flows from the shell 7 through the primary flow channel or connecting tube 17 ( see fig2 ) to the passageway 19 at the bottom of the valve housing 21 . from there , the air passes through the bottom of the valve housing 21 in fig2 and out the valve housing outlet at 25 into theflow path 27 which leads to the inlet 29 of the blower 33 . thereafter , the air passes through the main body 37 of the blower 33 to the outlet 39 and through the flow path 41 back into the valve housing 21 through the valve housing inlet 43 in the wall 45 . the air then flows out of the valve housing 21 through the valve housing outlet 47 . subsequently , as best seenin fig2 and 3 , the positive pressure air passes from the valve housing outlet 47 into a chamber 49 . this chamber 49 surrounds the right half of the valve housing 21 in fig3 and is created or defined by the enclosure formed between the back wall 45 and the cover member 51 with its dividers 53 ( see fig2 and 3 ). from chamber 49 , the air passes through the slots 55 ( see fig2 ) in the wall 45 and along the flow path 57 outside of the main body 37 of the blower 33 where it is vented to atmosphere through vent holes 59 . in this manner , a flow path from the shell 7 through the valve housing 21 to the blower 33 and back through the valve housing 21 to the atmosphere is thus created . referring again to fig2 the wall 45 containing the valve housing inlet 43 performs multiple functions . among them , the wall 45 functions at portion 45 &# 39 ; ( see fig4 and 5 ) as the back end wall for the valve housing21 . in this regard , the wall 45 at portion 45 &# 39 ; provides not only the valve housing inlet 43 as discussed above but also provides the valve housing outlet 25 at the bottom and back of the valve housing 21 . fig5 best illustrates this point in an exploded fashion wherein the wall portion 45 &# 39 ; with its openings 25 and 43 is shown below the exploded valving arrangement23 . the valving arrangement 23 as illustrated includes the valve housing 21and the single , one - piece valve member 2 ( which is explained in more detailbelow ). assembly of the exploded parts of fig5 then results in the assemblage in the positions of fig6 and 7 . the valving arrangement 23 of the present invention includes the valve housing 21 ( see fig5 ) and the single , one - piece valve member 2 . additionally , the portion 45 &# 39 ; of the end wall 45 as explained above forms the back end wall of the valve housing 21 when the valving arrangement 23 is assembled . when assembled , the valve member 2 is mounted as shown ( see fig5 - 7 ) within the valve housing 21 for rotation about the axis 4 . in one extreme position of the valve member 2 wherein it is rotated to its far clockwise position ( see fig1 - 7 ), a flow path is created from the shell 7 on the patient &# 39 ; s chest through the valve housing 21 to the blower 33 and back through the valve housing 21 to atmosphere . in this position , the full negative pressure of the blower 33 is applied to the chamber 15 in the shell 7 . in the other extreme position of valve member 2 wherein itis rotated to its far counterclockwise position ( skip ahead to fig1 and12 ), the condition exists in which the full positive pressure of the blower33 is being applied to the chamber 15 in the shell 7 . more specifically and referring first back to fig6 the valve housing 21 has a side opening 6 in its cylindrical wall 8 . with the cover 51 of fig2 in place , this opening 6 communicates with a chamber 49 &# 39 ; in essentially the same manner as explained above that the valve housing outlet 47 in fig2 and 3 communicates with chamber 49 . that is , the cover 51 with itsdividing walls 53 separates the space radially about the valve housing 21 into a left chamber 49 &# 39 ; and a right chamber 49 ( see fig7 ). the right chamber 49 as explained above is ultimately connected in fluid communication with the atmosphere through flow path 57 in fig2 . in a similar manner , chamber 49 &# 39 ; through slots 55 &# 39 ; and a flow path corresponding to 57 is also connected in fluid communication with the atmosphere . however , with the valve member 2 in its extreme , clockwise position of fig1 - 7 , the opening 6 in the valve housing 21 is closed to communication with any of the portals ( e . g ., passageway 19 , outlet 25 , or inlet 43 ) in the valve housing 21 . this is accomplished by the inclined , ramp member 10 of the valve member which is best seen in fig5 and 6 . inthis regard and with the valve member 2 in its extreme , clockwise position of fig6 and 7 , the ramp member 10 prevents or blocks any fluid communication through valve opening 6 with any of the other portals ( e . g ., 19 , 25 , or 43 ) of the valve housing 21 . the full negative pressure of the blower 33 can then be applied through passageway 19 to the chamber 15 in the shell 7 . conversely , with the valve member in its other extreme position ( i . e ., counterclockwise ) of fig1 and 12 , the flow path at 12 from the side opening 6 to the valve housing outlet 25 ( which leads to the inlet 29 of the blower 33 ) is not blocked by the ramp member 10 . consequently , the flow negative pressure draw of the blower 33 is connected to ambient or atmospheric air via the valve housing portal 25 , side opening 6 , chamber 49 &# 39 ;, and slots 55 &# 39 ;. at this position of valve member 2 , its elongated hole14 ( see fig5 and 11 ) connects with the valve housing inlet 43 . in this manner , the full positive pressure of the blower 33 is directed from the valve housing inlet 43 through the hole 14 past the upper side 16 ( see also fig1 ) of the ramp member 10 to the passageway 19 which in turn leads to the shell 7 . fig1 in this regard illustrates this flow throughthe valve housing 21 with the valve member 2 in this position from a view taken along line 13 -- 13 of fig1 . in a normal cycle of operation , the valve member 2 is moved between its extreme clockwise position of fig6 and 7 and a position approaching butnot actually reaching its extreme counterclockwise position of fig1 and12 . that is , the most common mode of operation of the negative pressure ventilator 3 is intended to produce a pressure cycle in the chamber 15 of shell 7 such as shown in fig1 . in such a cycle , the pressure in the shell 7 changes between negative and positive , preferably abruptly to compensate for the resistance of the patient &# 39 ; s chest wall . also , the negative pressure side is preferably smaller in time ( e . g ., 40 percent of each cycle ) and larger in relative pressure ( e . g ., negative 30 centimetersof water ) than the positive pressure side ( e . g ., 60 percent in time and positive 5 centimeters of water ). in this manner , the pressure in the shell chamber 15 is reduced to 30 centimeters of water in our example ( i . e ., from point a to point b in fig1 ) causing the patient to breathe in or inhale . this negative pressure is then gradually relieved to zero ( i . e ., from point b to point c of fig1 ) allowing the patient &# 39 ; s chest tofall under its own weight and the patient to breathe out or exhale . additionally , the cycle then continues to include a slight rise to a smallpositive pressure in the chamber 15 of 5 centimeters of water in our example ( i . e ., from point c to point d in fig1 ) to slightly compress the patient &# 39 ; s chest to aid his exhalation . thereafter , the positive pressure is reduced to zero ( i . e ., from point d to point a &# 39 ; in fig1 ) tobegin a new cycle . for clarity and in reference to the positioning of the rotatably mounted valve member 2 , this normal cycle of operation is perhaps easier to understand , describe , and illustrate by following the cycle in fig1 from trough to trough ( i . e ., line t -- t in fig1 ). more specifically , at the bottom of the trough , the valve member 2 is in its extreme clockwise position of fig6 and 7 . in this position , the full negative pressure ofthe air blower 33 is being applied through passageway 19 and the connectingor primary flow channel 17 to the chamber 15 in shell 7 . from there , the valve member 2 is rotated slightly counterclockwise to the position of fig8 . in this position , the ramp member 10 which completely sealed off portal 25 from chamber 49 &# 39 ; in fig7 has been moved to slightly uncover the valve housing outlet 25 ( which in turn leads to the inlet 29 of the blower 33 ). the negative draw of the blower 33 then draws both from passageway 19 leading to the patient &# 39 ; s shell 7 and from atmospheric air via side opening 6 , chamber 49 &# 39 ;, and slots 55 &# 39 ; leading to ambient air . thenegative pressure in the shell 7 thus begins to be relieved ( e . g ., to 15 centimeters of water -- see fig1 ). continued counterclockwise rotation ofvalve member 2 to fig9 produces a zero draw through passageway 19 leadingto the patient &# 39 ; s shell 7 . in the position of fig9 the negative draw of the blower 33 is completely drawn from ambient air through chamber 49 &# 39 ; andportal 25 . additionally , the exhaust or positive pressure discharged from the blower 33 is directed through valve housing outlet 47 to chamber 49 toatmosphere . the cycle to the shell 7 is thus at a dead spot ( i . e ., zero , nonegative or positive pressure being applied through passageway 19 into shell 7 ). further rotation of valve member 2 counterclockwise to fig1 then slightly connects part of the positive pressure discharge at 43 from the blower 33 to the elongated hole 14 of the valve member 2 . a slight positive pressure ( e . g ., 5 centimeters of water ) is thus driven or directed to passageway 19 leading to the shell 7 to assist the patient in exhalation . the valve member 2 is then rotated back clockwise in essentially a mirror image manner to again reach a negative pressure trough in our example of 30 centimeters of water . in normal operation , fig1 is as far counterclockwise as the valve member2 is rotated in a cycle . this is true because patients rarely need the fullpositive pressure of the blower 33 ( e . g ., 30 centimeters of water ) applied to them . yet , the capacity to do so is built into the valving arrangement 23 of the present invention . that is , if desired , the valve member 2 can be moved to the extreme counterclockwise position of fig1 and 12 wherein the valve housing outlet 47 is completely blocked and all air fromthe blower 33 is directed or driven through hole 14 in the valve member 2 into the passageway 19 leading to the patient &# 39 ; s shell 7 . this position could be used to apply the full positive pressure to the patient or simplyto rapidly relieve the pressure or shell by bringing it very rapidly back to zero . for additional clarity , the operation of the valving arrangement 23 of the present invention has been illustrated in schematic form in fig1 - 20 . fig1 is a two - dimensional representation of the present invention . as shown in it , the connecting tube or primary flow channel 17 to the patientis connected to the passageway 19 of the valve housing 21 . this passageway 19 for clarity has been described as a separate part to give a reference point of where the primary flow channel 17 enters the valve housing 21 . however , passageway 19 and primary flow channel 17 could simply be portions of the same or continuous tube . in any event , the valve housing 21 then has a first outlet 25 in the schematic of fig1 leading through a first flow path 27 to the blower inlet 29 . the positive pressure air from the blower outlet 39 is directed through the second flow path 41 to the first inlet 43 of the valve housing 21 . additionally , valve housing 21has a second outlet 47 connected via chamber 49 and slots 55 to ambient air . valve housing 21 in the schematic of fig1 also has a second inlet ( i . e ., side opening 6 in the valve housing 21 ) connected via chamber 49 &# 39 ; and slots 55 &# 39 ; to ambient air . in operation in the two - dimensional schematics of fig1 - 20 , the valving arrangement 23 can first be positioned as shown in fig1 to draw 100 % ofthe negative pressure of the blower 33 through the primary flow channel 17 leading to the patient &# 39 ; s shell 7 . in this position of fig1 , 100 % of thepositive pressure air from the blower 33 is exhausted through the second valve housing outlet 47 to atmosphere . fig1 thus corresponds to fig1 - 7 . thereafter , the valve member 2 can be rotated counterclockwise to thesecond position of fig8 which is schematically shown in fig1 . in this position , air is drawn both from the primary flow channel 17 and the second valve housing inlet 6 . in this regard , the valve member 2 can be rotated or positioned to draw any desired proportion of air ( e . g ., 75 % from the primary flow channel 17 and 25 % from opening 6 or whatever ) but it is intended to draw 100 % of the demand of the blower 33 only from thesetwo sources ( i . e ., x % from 17 and 100 %- x % from 6 ). the valve member 2 is thereafter rotated to its neutral or dead spot of fig9 and 18 with the blower 33 drawing 100 % of its negative flow from atmosphere through the second valve housing inlet 6 and discharging 100 % of its positive flow to atmosphere through the second valving outlet 47 . in this position , the valving arrangement 23 completely bypasses any communication with the patient through the primary flow channel 17 and the air for the blower 33 is drawn entirely from ambient air and the air from the blower 33 is discharged entirely to the ambient air . this neutral position with its by - pass flow is desirable as the blower 33 is not working against a dead space . consequently , the by - pass flow in the neutral position helps to keep the blower 33 cool . continuing rotation of valve member 2 to the third position of fig1 ( schematic fig1 ) then serves to apply some positive pressure ( e . g ., y % or enough to apply 5 centimeters of water ) to the shell 7 . however , the bulk ( 100 %- y %) of the positive pressure air in this position is being vented through the second valve housing outlet 47 to atmosphere . this movement through fig1 - 19 is then essentially reversed to complete a cycle . schematic fig2 illustrates the capacity of the valving arrangement 23 toposition valve number 2 in its extreme counterclockwise position . this corresponds to fig1 and 12 and applies to the patient 100 % of the ambient air being drawn through the second valve housing inlet 6 into the blower 33 . as discussed above , however , this is not a normal step in the operation of the present invention but is available if desired and if required by the particular condition of the patient 1 . for example , it maybe desirable in certain cases to abruptly relieve the negative pressure to zero by infusing full positive pressure to the shell chamber 15 . schematic fig1 - 20 and in particular fig1 also illustrates the preferred manner of operation of the valving arrangement 23 . in it , the periodic delivery or flow of air is preferably managed without ever connecting the negative and positive sides of the blower 33 at the same time to the shell 7 on the patient &# 39 ; s chest . otherwise , the opposing pressures would work against one another trying both to inflate and deflate the patient &# 39 ; s lungs at the same time . this is accomplished in the preferred embodiment by having a specific position of valve 2 ( i . e ., figs . 9 and 18 ) in which structure positively prevents the undesirable application of both positive and negative pressure to the patient 1 . thus , the valving arrangement 23 of the present invention offers several distinct advantages over prior art devices . for example , the blower 33 in the system can be set at a single speed ( e . g ., 5 , 000 rpm &# 39 ; s or a single speed blower 33 can simply be used ) and the valving arrangement 23 can then deliver either positive or negative pressure to the patient 1 anywhere from zero to the maximum blower pressure ( positive and negative ). additionally , if a servo motor is used , for example , this pressure deliveryto the patient 1 can be infinitely adjusted by the valving arrangement 23 . further , the blower 33 can be operated at a slightly higher speed ( e . g ., 7 , 000 rpm &# 39 ; s ) to create a maximum pressure ( e . g ., 35 centimeters of water ) in excess of the maximum pressure ( e . g ., 30 centimeters of water ) normallyapplied to the shell chamber 15 . in this mode of operation , the valve member 2 would not be moved to its extreme clockwise position during a normal cycle and would essentially operate between a cycle of just fig1 and 19 . the disadvantage of running the blower 33 at such a higher speed and pressure is for the most part just increased noise . the advantage gained , however , is that the valve arrangement 23 can then compensate for leakage out of the shell chamber 15 ( e . g ., if the patient 1moves or if the seal at the edge 11 changes ). in this regard , for example , if the desired maximum negative pressure in the shell chamber 15 is negative 30 centimeters of water and the blower 33is run at negative 35 centimeters of water , the valve member 2 may then be rotated only to the position of fig1 to draw the pressure down in shellchamber 15 to 30 centimeters of water . however , if a leak occurs or the size of the leak increases , the valve member 2 can simply be rotated farther clockwise to connect the tube 17 with , for example , 32 or the full35 centimeters of water from the blower 33 . this will then compensate for the leakage and maintain the maximum desired draw of 30 centimeters of water over the patient &# 39 ; s chest in the shell chamber 15 as sensed by sensor38 . if the leakage or change in leakage is sudden , the valving arrangement 23 of the present invention can adjust to it virtually instantaneously by a simple movement of valve member 2 . in contrast , prior single speed blowers normally cannot compensate for such leaks or must use separate valves or bleeders and prior adjustable speed blowers usually compensate for such leaks by reving up the blower to a higher speed which can often take several minutes . the valving arrangement 23 can therefore be used to enhance the operation of such adjustable blowers as well as less expensive , single speed blowers . the valve housing 21 of the valving arrangement 23 as shown in fig5 has asubstantially cylindrical , curved wall 8 extending about and along the axis4 . the wall 8 has interior exterior , curved sides 18 and 20 ( see fig7 ) that also extend substantially about and along the axis 4 between the valve housing end walls 45 &# 39 ; and 22 ( see fig5 ). the valve housing end walls 45 &# 39 ; and 22 as illustrated extend substantially perpendicular to the axis 4 . the valve member 2 , in turn , is positioned within valve housing 21between the end walls 45 &# 39 ; and 22 ; and , the passageway 19 as illustrated extends through the front end wall 22 ( see fig5 ). in the preferred embodiment , the first outlet 25 and first inlet 43 of the valve housing 21pass through the back end wall at 45 &# 39 ;; and , the second inlet 6 and second outlet 47 respectively pass through the cylindrical wall 8 of the valve housing 21 . the valve member 2 as disclosed above includes ramp member 10 which is inclined to the axis 4 . ramp member 10 extends between the two end walls 45 &# 39 ; and 22 and essentially divides the valving arrangement 23 internally into two chambers 24 and 26 ( see fig5 ). chamber 24 is illustrated as always being to the left or clockwise from the dividing ramp member 10 in the figures and chamber 26 is illustrated as always being to the right or counterclockwise from the ramp member 10 in the figures . curved portion 28of valve member 2 ( see fig5 ) also helps to define chamber 26 and further serves with portion 30 to control and valve the valve housing outlet 47 ( see fig1 and 11 ). in operation as discussed above , chamber 24 is always in fluid communication with the second inlet 6 of the valve housing21 at least in the three valve positions of fig7 , and 10 and corresponding schematic fig1 , 17 , and 19 . in all of these last - mentioned valve positions as shown , the ramp member 10 prevents fluidcommunication within the valve housing 21 between the valve and inlets 6 and 43 . the other chamber 26 when the valve member 2 is in the third position of fig1 ( see also schematic fig1 ) is always in fluid communication with the first inlet 43 of the valve housing 21 . in the neutral or dead spot position of fig9 and 18 , chamber 26 is isolated from fluid communication with either portal 25 or 43 and in this position , neither positive nor negative pressure air is directed to the passageway 19leading to the shell 7 . that is , in this neutral position , the primary flowchannel 17 to the patient 1 is closed . the unique structure of the valving arrangement 23 allows the operator to easily program , adjust , and vary the operation of the system to meet a particular patient &# 39 ; s needs . this is primarily a consequence of the fact that the operation of the system is essentially controlled by simply manipulating a single , one - piece valve member 2 . in this light , for example , the cycle time and pressure limits of fig4 can be set by simplyadjusting knobs 32 of the programmable control unit 34 on the front of the negative pressure ventilator 3 of fig1 and 2 . the pressure sensor 36 with its feedback sensor line 38 of fig1 and 2 then insures that the desired pressure settings at the chamber 15 are met . in the example of fig1 , the desired pressure limits to the patient are set between negative 30 centimeters of water and positive 5 centimeters of water . the turbine 31 is then run by motor 35 ( which is also controlled by unit 34 ) to a speed ( e . g ., 5 , 000 rpm ) that will generate negative 30 centimeters ofwater in chamber 15 ( as sensed by sensor 36 through its sensor line 38 ). this is preferably at the extreme clockwise position of the valve 2 ( i . e ., position of fig1 - 7 ) which connects the full negative draw of the blower 33 to the shell 7 . as a practical matter , the blower 33 is then being operated as quietly and cool as possible since it operating at the lowest speed to achieve the desired , maximum pressure . the speed and positioning of how the valve member 2 is then moved counterclockwise through fig8 - 10 is controlled in response to operationof the drive 40 ( see fig2 and 21 ). the drive 40 ( e . g ., servo motor ) as shown includes a simple belt and pulley arrangement 42 wherein valve member 2 is selectively rotated about axis 4 . this is all controlled by the control unit 34 of fig2 in cooperation with the feedback sensor 36 which seeks to create the pre - set , desired cycle of fig1 in as smooth amanner as possible . this simplicity of operation and the infinite adjustability of valve member 2 enable the cycle or cycles to be set to include any number of variations . these may include such desirable features as a sigh function ( e . g ., a relatively large breath over a longerthan normal time at a negative pressure greater than the negative 30 centimeters of water in our example ). such a sigh function can be accomplished by simply increasing the turbine speed from its normal rate of , for example , 5 , 000 rpm to 13 , 000 rpm and appropriately manipulating valve member 2 to its extreme clockwise position of fig1 - 7 . the cooperation between the sensor 36 with its feedback line 38 and the control unit 34 also automatically adjusts for leaks in the system ( eithercontinuous or temporary ones ). it can do this by ramping up the turbine 31 to whatever speed is necessary to achieve the desired , maximum negative pressure at the chamber 15 in shell 7 . the system also quickly adjusts to any size shell or other hard or soft covering ( e . g ., pancho , full body suit ) on the patient . that is , the system automatically operates to createthe pre - set pressure differentials in the shell 7 or other covering regardless of its volume . the operation can also include breaths in which more positive pressure is directed into the shell 7 to more fully compressthe lungs . this can be accomplished by either rotating the valve member 2 counterclockwise beyond fig1 to allow increased pressure to develop in shell 7 . a cycle of operation could additionally be set at less than completely breathing for the patient so that he would have to work his muscles somewhat . all of this flexibility is easily achieved by the systemof the present invention primarily because of the structure of the single , one - piece valve member 2 which can be easily , quickly , and accurately adjusted and controlled to move among infinitely variable positions . while several embodiments of the invention have been shown and described indetail , it is to be understood that various modifications and changes couldbe made to them without departing from the scope of the invention .

a valving arrangement primarily designed and intended for use in a negative pressure ventilator system . the valve arrangement has a uniquely designed valve that enables the ventilator to perform a number of operations simply by manipulation of a single , one - piece valve member within a valve housing . the valve housing has multiple inlets and outlets and the valve member can be rotated within it to present a variety of pressure cycles to the patient . in the preferred manner of operation , the present invention selectively applies negative and positive pressures to the patient during different portions of the cycle . in this regard , the valve member is structured so that negative and positive pressures are never at the same time delivered to the patient . the valve arrangement is compact and lightweight . it is also functionally superior to prior designs that can only supply whatever pressure is being created by the blower . in contrast , the valve of the present invention can create pressures from zero to the maximum blower pressures by positioning the valve to selectively split the blower pressure to ambient air .

with reference to the figures . in the preferred embodiment , buttons 106 have an upper barrel , 107 , a lower barrel , 108 , and a cap 109 . the cross - section of the upper barrel 107 is generally rectangular as is the cross - section of the lower barrel 108 . furthermore , the width w of the lower barrel 108 is slightly larger than the width w of the upper barrel 107 . rubber stamp , 112 is attached by any convenient means to the bottom end of lower barrel 108 . the top surface 102 of hollow rectangular base 101 contains a 5 × 5 matrix of rectangular holes 103 . the dimensions of the holes are such that the upper barrel 107 of button 106 will slide therethrough with little play . the spacing of the holes is such that they will be in alignment with each location on a bingo card . plate 104 , which is located parallel to the top surface 102 of base 101 , may be affixed to the inside of the base by any convenient means . plate 104 also has a 5 × 5 matrix of holes 105 contained within its surface . the dimensions of said holes are such that the lower barrel 108 of button 106 will slide therethrough with little play . the location of holes 105 is such that they are in alignment with each location on a bingo card . the width of rectangular set - reset plate 113 , which may be made of any material which is convenient , such as a plastic material , is slightly less than the width of the interior of top surface 102 of base 101 , and the length of plate 113 is slightly less than the length of the interior of surface 102 so that plate 113 may be attached to surface 102 in a manner which permits plate 113 to slide back and forth in the lengthwise direction . in the preferred embodiment rivets 114 , which pass through slots 115 , provide the slidable attachment means between surface 102 and plate 113 . extension 116 of plate 113 protrudes from the assembly and provides the means for moving plate 113 to the left at the urging of a user &# 39 ; s hand . upper and lower spring means 117 , one of which is shown in figure 4 , is attached , on one end , to ear 118 on base 101 , and attached , on the other end to ear 119 on plate 113 . spring means 117 urges plate 113 to the right . plate 113 contains a 5 × 5 matrix of rectangular holes 120 which are in alignment with holes 103 and 105 so that buttons 106 may pass therethrough . the depth of hole 120 ( fig6 ) is the same as the depth of hole 103 . ( fig1 ). each position in the matrix has a tab 121 which extends into hole 120 and provides the detent means for locking button 106 in an extended position . the width of holes 120 is such that the upper barrel 107 of button 106 will clear the end of tab 121 when the set - reset plate ( 113 ) is pushed to the left . tab 121 contains transverse groove 122 . ( fig2 ). the cross - section of the grove may be rectangular , triangular or any other convenient shape so long as the material of the tab is thinned by the groove thereby creating a hinge at the groove which allows tab 121 to pivot and slide up ramp 110 when a button is depressed . when button 106 is depressed , tab 121 will slide up ramp 110 , and then up the body of button 106 until tab 121 enters notch 111 . notch 111 may have a rectangular cross - section and extend transversely entirely across upper barrel 107 of button 106 or may be a cavity formed in barrel 107 . this form of notch is shown in fig3 . in the latter case , the material remaining on either side of the cavity provides a bearing surface which allows button 106 to move freely within holes 103 . ramp 110 , which extends transversely across upper barrel 107 of button 106 , may have any convenient slope so long as tab 121 is caused to rotate about hinge point 122 . spring means 123 surrounds upper barrel 107 and is compressed between cap 109 and surface 102 so that buttons 106 are urged upward at all times . other forms of this invention are within the contemplation of the inventor . for example , button 106 could comprise several axially aligned cylinders . this form of the button is shown in fig5 . a shown therein , upper barrel 107 has a larger diameter than lower barrel 106 , ramp 110 has been eliminated , upper surface 102 of base 101 has been thickened to provide a bearing surface which surrounds notch 111 . spring means 123 has been moved so that it surrounds lower barrel 106 . spring retaining clip 124 has been added to hold spring 123 in a compressed state and stop clip 125 has been added to prevent button 106 from moving upward beyond a desired home position . other forms of set - reset plate are also within the contemplation of the inventor . such as for example , making detent tabs 121 of spring steel so that groove 122 may be eliminated , or making set reset plate 113 entirely of spring steel .

a bingo card stamping device comprising a 5 × 5 matrix of marker buttons , having individual stamping means attached to the bottom of each button , and which can be set in any pattern desired by the user whereby the user may stamp the pattern on a bingo card .

the following detailed description illustrates the invention by way of example and not by way of limitation . the description clearly enables one skilled in the art to make and use the invention , describes several embodiments , adaptations , variations , alternatives , and uses of the invention , including what is presently believed to be the best mode of carrying out the invention . referring now to fig1 through fig3 reference numeral 10 illustrates one embodiment of the hand - held sub - microvolt electrical signal measurement device of the present invention . the measurement device 10 includes an enclosure 12 , which in the preferred embodiment , and for purposes of illustration and not for limitation , measures 7¼ ″ long by 3¾ ″ wide by 1½ ″ deep . it is important to note that the device 10 can be carried by the user without compromise , and truly represents a portable hand - held device having full functionality as described below . the device 10 includes a keyboard 14 and an lcd display 16 . one or more led indicators are optionally included , such as an led pass / refer indicator 18 , and an led ac charging indicator 20 . again , by way of illustration and not by limitation , it should be noted that the lcd display 16 measures , in the preferred embodiment , approximately , 2 ″ by 3⅜ ″. the measurement is not necessarily important , except to show that the lcd display 16 is fully functional for a user , and the device 10 can operate independently of any other computer system . in the embodiment illustrated , the enclosure 12 also houses an infrared port 22 , and a compatible rs - 232 port 24 , a probe connection 26 suitable for use with an input probe 28 , such as an ear probe , and an interface 30 for a plurality of output electrodes 32 . the electrodes 32 are shown attached to a conventional carrier 34 . probe 28 is conventional and is not described in detail . suitable probes , such as ear probes , are commercially available from etymotic research , part nos . er - 10c , er - 10d , and gsi 2002 - 3250 , for example . referring now to fig4 a block diagram view of the hand - held sub - microvolt electrical signal measurement device 10 is shown and described . preferably , the system shown in fig4 is manufactured on a single printed circuit board , with mixed signal design for both analog and digital operation . the device 10 preferably is low powered , and generally operates at 3 . 3 volts , except for the lcd display 16 and some low power portions of the analog circuitry employed with the device 10 . to reduce undesired signal interference , it has been found that it is preferable that all analog and digital circuit components share a common electrical ground point within the device 10 . a suitable micro - processor 40 is the control for the device 10 . in the preferred embodiment illustrated , the processor 40 is a motorola model no . 56303 digital signal processor , however , those of ordinary skill in the art will recognize that any suitable micro - processor or micro - controller having sufficient computational power and speed may be utilized . all signal processing functions described hereinafter are performed by the processor 40 , as well as the control of all input and output functions of the device 10 . in addition , graphic functions , user interface functions , data storage functions , and other device functionality are controlled by the processor 40 . in conventional design logic , the digital signal processor 40 is used for signal processing , and a separate micro - controller is used for device control . in device 10 , the digital signal processor 40 performs the functions of the separate micro - controller in addition to signal processing , eliminating the requirement of a separate micro - controller , resulting in substantial savings in circuit board space , manufacturing cost and operational power consumption . to reduce undesired signal interference during the data - acquisition phase of operations , the processor 40 in device 10 is either shut - down or switched to a “ sleep ” mode during data collection operations , which can be carried out independently of the processor 40 . further reduction in external signal noise is achieved by the execution of software in data and program memory 40 m internal to processor 40 , thereby eliminating external bus access signal noise . a memory subsystem 42 is operatively connected to the processor 40 . the memory subsystem 42 includes a random access memory ( ram ) 42 a for storing intermediate results and holding temporary variables , and a flash memory 42 b for storing non - volatile , electrically programmable variables , test result data and system configuration information . in the embodiment illustrated , the flash memory 42 b is substantially oversized , enable the device 10 to accommodate several hundred data records , as well as multiple configurations files . a memory mapped input / output device 44 is operatively connected to the memory subsystem 42 and to the digital signal processor 40 . the memory mapped input / output 44 in turn is operatively connected to the lcd display 16 , the keyboard 14 , an output led indicator 18 and a real time clock 46 . the lcd display 16 provides the user with a display array preferably having a minimum size of 128 × 256 pixels . a display array of this size is sufficient to present full waveforms of signal tests conducted by the device 10 . the device 10 enables the lcd 16 to present signal information to a user graphically in real time on the device 10 itself , complemented with textual and numeric information about the quality of the data , signal amplitudes , signal frequency , noise floors and other related signal information . the keyboard 14 preferably is a membrane switch keyboard , which incorporates only the minimum keys necessary for operation of the device 10 . all keys are programmable , except for an on / off key 15 . the real - time clock 46 is operatively connected to the processor 40 through the memory mapped input / output device 44 . the real - time clock 46 enables the processor 40 to provide a time stamp for each data collection or test performed . the output led 18 is used to convey test results to non - trained users to avoid confusion or misinterpretation of the lcd graphics display 16 . for example , the processor 40 may be programmed to illuminate the output led 18 when a set of predetermined input criteria , such as a signal strength at or above a minimum value . the output led 18 further allows the use of the device 10 in low light areas , where the lcd display 16 may be difficult to read or interpret . the plurality of analog to digital / digital to analog coder / decoders 48 ( codecs 48 ) are operatively connected to the signal processor 40 along a dedicated serial link . as will be appreciated by those skilled in the art , the codecs 48 are special integrated circuit chips that perform analog to digital and digital to analog conversion . the codecs 48 in turn are operatively associated with a one or more input / output devices interfaces 52 , which provide the functionality of the device 10 under control of the processor 40 . preferably , the digital signals generated and received by codecs 48 have 20 - bit resolution for both analog to digital and digital to analog conversions . in one embodiment of the present invention 10 , input / output interface 52 includes four input channels and four output channels adapted for sub - microvolt electrical signals . as shown in fig5 the input / output interface 52 consists of a plurality of analog signal processing chips , not shown individually , which filter and amplify the signals received from a number of discrete input channels 32 . specifically , signals received from each input channel 32 are routed through an electrical insulator circuit 52 a , consisting of a plurality of metal oxide varistors , resistors , and capacitors , which function as surge arrestors to isolate the input channels 32 from any dangerous electrical currents or voltages . the insulator circuit 52 a functions to replace conventional insulator circuits which utilize optical signal pathways , thereby eliminating the associated signal noise resulting from the conversion between electrical signals and optical signals . signals from the input channels 32 are then routed through a switching network 52 b , wherein an individual signal is automatically selected and passed to a capacitive coupled differential operational amplifier 52 c having high gain . a common mode cancellation amplifier 52 d is included in the input / output interface 52 to further reduce signal noise levels . the resulting amplified signal is then routed to the codecs 48 . in addition to selecting an individual signal from the input channels 32 , the switching network 52 b permits a variety of signal measurements on the input channels 32 to be carried out using the same differential operational amplifier 52 c , by altering the amplifier gain setting . returning to fig4 a mode configuration system 54 , a reset watchdog system 56 , a clock crystal 58 , a power supply 60 , preferably a nickel - metal hydride battery , and a battery charger 62 all are also positioned within the enclosure 12 and operatively connected to the processor 40 . while each of these blocks is required for operation of the device 10 , they are standard in nature and are not described in detail . the processor 40 has an input - output channel 64 , which preferably includes an infrared connection 22 , a fiber - optic connection 23 and an isolated rs - 232 interface 24 . the device 10 can communicate with any infrared compatible or rs - 232 compatible personal computer , printer , or other digital device ( not shown ) for data transmission . data transmission may include test subject information , configuration data for the signal processor 40 , or software program updates for storage in the memory subsystem 42 . a audio output 66 in the form of a buzzer also is provided . the audio output 66 provides an audio feedback to the user for keyboard actions and an audio indication for error conditions . a serial port 68 also is operatively connected to the processor 40 . the serial port 68 is utilized to provide direct programming of the processor 40 from a personal computer , for example , and is intended for use only for initial software download and major software program upgrades of the processor 40 . referring now to fig6 a block diagram view of one embodiment of the device 10 , configured for use as an auditory screening device 200 , is shown and described . the device 200 contains oae and abr simulator capabilities in a single , hand - held package . preferably , the system shown in fig6 is manufactured on a single printed circuit board , with mixed signal design for both analog and digital operation . the device 200 preferably is low powered , and generally operates at 3 . 3 volts , except for the lcd display 216 and some low power portions of the analog circuitry employed with the device 200 . to reduce undesired signal interference , it has been found that it is preferable that all analog and digital circuit components share a common electrical ground point within the device 200 . a digital signal processor 240 is the control for the device 200 . in the preferred embodiment illustrated , the processor 240 is a motorola model no . 56303 dsp . all signal processing functions described hereinafter are performed by the processor 240 , as well as the control of all input and output functions of the device 200 . in addition , the graphic functions , user interface , patient data storage functions and other device functionality are controlled by the processor 240 . in conventional design logic , the digital signal processor 240 is used for signal processing , and a separate micro controller is used for device control . in device 200 , the digital signal processor 240 performs the functions of the separate micro controller in addition to signal processing , eliminating the requirement of a separate microprocessor , resulting in substantial savings in circuit board space , manufacturing cost and operational power consumption . to reduce undesired signal interference during the data - acquisition phase of operations , the processor 240 in device 200 is either shut - down or switched to a “ sleep ” mode during data collection operations , which can be carried out independently of the processor 240 . further reduction in external signal noise is achieved by the execution of software in data and program memory 240 m internal to processor 240 , thereby eliminating external bus access signal noise . a memory subsystem 242 is operatively connected to the processor 240 . the memory subsystem 242 includes a random access memory ( ram ) 242 a for storing intermediate results and holding temporary variables , and a flash memory 242 b for storing non - volatile , electrically programmable variables , patient data , and configuration information . in the embodiment illustrated , the flash memory 242 b is substantially oversized , enable the device 200 to accommodate several hundred full patient records , as well as multiple configurations files . a memory mapped input / output device 244 is operatively connected to the memory subsystem 242 and to the digital signal processor 240 . the memory mapped input / output 244 in turn is operatively connected to the lcd display 216 , the keyboard 214 , the pass / referral led indicator 218 and a real time clock 246 . the lcd display 216 provides the user with a display array preferably having a minimum size of 128 × 256 pixels . a display array of this size is sufficient to present full waveforms of audiometric tests conducted by the device 200 . the device 200 enables the lcd 216 to present signal information to a user graphically in real time on the device 200 itself , complemented with textual and numeric information about the quality of the data , signal amplitudes , signal frequency , noise floors and other related signal information . the keyboard 214 preferably is a membrane switch keyboard , which incorporates only the minimum keys necessary for operation of the device 200 . all keys are programmable , except for an on / off key ( not shown ). the real - time clock 246 is operatively connected to the processor 240 through the memory mapped input / output device 244 . the real - time clock 246 enables the processor 240 to provide a time stamp for each patient and test performed , as well as providing time signals for internal operation of the device 200 . the led pass / refer diode 218 is used to convey test results to non - trained users , namely a nurse as opposed to an audiologist or otolaryngologist . use of the led 218 avoids confusion or misinterpretation of the lcd graphics display 216 , and allows use of the device 200 in low light areas , where the lcd display 216 may be difficult to interpret . the plurality of analog to digital / digital to analog coder / decoders 248 ( codecs 248 ) are operatively connected to the signal processor 240 along a dedicated serial link . as will be appreciated by those skilled in the art , the codecs 248 are special integrated circuit chips that perform analog to digital and digital to analog conversion . the codecs 248 in turn are operatively associated with a plurality of input / output devices , which provide the functionality of the device 200 under control of the processor 240 . an otoacoustic emission interface 250 ( dpoae i / f ) is operatively connected to the signal processor 240 through the associated codecs 248 . the otoacoustic emission interface 250 preferably is a low noise , differential analog circuit with high common mode noise rejection . as shown in fig7 the otoacoustic emission interface 250 is intended to drive two sound transducers 251 r , 251 l through a pair of differential operational amplifiers 250 a , 250 b to produce a variety of signals , from pure tones at various frequencies to chirps , clicks , sine waveforms , etc . the otoacoustic emission interface 250 can present tones at standard sound pressure levels . the device employed with the otoacoustic emission interface 250 includes a microphone 251 m , also inserted in the ear canal , which collects signals coming back from the ear , and provides sufficient linear amplification through a dual - stage amplifier 250 c to present the signals to the codecs 248 . the transducers and microphone interface circuits include a plurality of electrostatic discharge diodes and induction coils to provide electrical shock protection 250 d . in various embodiments of this invention , the otoacoustic emission interface 250 also can be used for otoreflectance measurements for assessing some middle ear conditions . the abr interface 252 , shown in fig8 consists of a plurality of analog signal processing chips , not shown individually , which filter and amplify the signals received from the scalp of a subject via electrode wires 232 . specifically , signals received from each of the individual electrodes 234 are routed through an electrical insulator circuit 252 a , consisting of a plurality of metal oxide varistors , resistors , and capacitors , which functions to isolate the electrodes 234 from any dangerous electrical currents or voltages . the insulator circuit 252 a functions to replace conventional insulator circuits which utilize optical signal pathways , thereby eliminating the associated signal noise resulting from the conversion between electrical signals and optical signals . signals from the electrodes 234 are then routed through a switching network 252 b , wherein an individual signal is automatically selected and passed to a differential operational amplifier 252 c . a common mode cancellation amplifier 252 d is included in the abr interface 252 to further reduce signal noise levels . the resulting amplified signal is then routed to the codecs 248 . in this mode of operation , the ear is presented with a repeated acoustic stimuli that cause neurons to fire beginning with the eighth cranial nerve and sequentially through neurons in the auditory pathways in the central nervous system from the brainstem to the cortex . through the mechanism of volume conduction , the electrical potentials generated from these neuronal firings can be detected by the electrodes 232 on the surface of the skin . an additional function of the abr interface 252 is to provide automated impedance check of the placement of electrodes 232 . once the electrodes 232 are in place , a small current is injected through the electrodes 232 into the scalp of the subject , and the impedance between electrodes 232 is measured . in addition to selecting an individual signal from the electrodes 234 , the switching network 252 b permits impedance measurements of the electrodes 234 to be carried out using the same differential operational amplifier 252 c , by altering the amplifier gain setting . impedance can be varied by placement of the electrodes . once the impedance is within the predetermined range for operation , abr signal connection can begin . it is important to note that impedance checking can be accomplished without unplugging the electrodes . that is to say , checking is automatic . returning to fig6 a mode configuration system 254 , a reset watchdog system 256 , a clock crystal 258 , a power supply 260 , preferably a nickel - metal hydride battery , and a battery charger 262 all are operatively connected to the processor 240 . while each of these blocks is required for operation of the device 200 , they are standard in nature and are not described in detail . the processor 240 has an input - output channel 264 , which preferably includes infrared connection 22 , fiber - optic connection 23 and isolated rs - 232 interface 24 . the device 200 can communicate with any infrared compatible or rs - 232 compatible personal computer , printer , or other digital device ( not shown ) for data transmission . data transmission may include patient information , configuration data for the signal processor 240 , or software program updates for storage in the memory subsystem 242 . a audio output 266 in the form of a buzzer may be provided . the audio output 266 provides an audio feedback to the user for keyboard actions and an audio indication for error conditions . a serial port 268 also is operatively connected to the processor 240 . the serial port 268 is utilized to provide direct programming of the processor 240 from a personal computer , for example , and is intended for use only for initial software download and major software program upgrades of the processor 240 . as an auditory screening device 200 , the present invention utilizes a auditory phenomena known as a distortion product otoacoustic emission ( dpoae ). a dpoae is a tone generated by a normal ear in response to the application of two external tones . when two tones , f 1 and f 2 are applied to an ear , the normal non - linear outer hair cells generate a third tone f dp , which is called a distortion product . the distortion product f dp then propagates from the outer hair cells back to the ear canal where it is emitted . the level of the dpoae can be used as a measure of outer hair cell function . if the outer hair cell system is absent or otherwise not functioning properly , the non - linearity will be absent or reduced and the f dp will either not be generated or generated at a lower than expected level . the measured dpoae is highly dependent upon the specific tones that invoke it . the frequencies of f 1 and f 2 , and their respective levels in the ear canal , l 1 and l 2 must be controlled precisely . under known signal conditions , the largest distortion product f dp is generated at a very specific frequency ( f dp = 2f 1 − f 2 ), and level l dp . comparison of the level of l dp with known values from individuals with normal outer hair cell systems forms the basis of the decision of whether the patient either passed the screening , illuminating the pass / refer led 218 , or requires a referral for a more complete diagnostic testing . signals other than pure tones can be presented to the ear , which will also evoke an auditory response from the ear , such as clicks , chirps , etc . the dpoae response is used with the auditory screening device 200 as an example of one such input . other auditory stimuli generating an auditory response would be processed by the auditory screening device 200 the same way as the dpoae . as shown in fig6 during operation , the processor 240 sends a filtered output signal from a filter 268 through the digital to analog converter portions 270 of the codecs 248 . the output signals are then routed through amplifier components 250 a and 250 b in the dpoae interface 250 and transmitted to the output components 251 r and 251 l of the ear probe 228 utilized in conjunction with the device 200 . as shown in fig7 the ear probe 228 includes a microphone 270 which returns signals through a second amplifier 250 c in the dpoae interface 250 . the amplified analog signal is routed through an analog - to - digital converter 280 in the codecs 248 , and conveyed to the processor 240 . in the processor 240 , the incoming analog signal is sampled using a frame buffer 282 . the size of each new frame in the frame buffer 282 is calculated to be an integer number of samples of the two primary tones at frequencies f 1 and f 2 and also , an integer number of samples of the otoacoustic tone produced by the ear at f dp . this is a critical step to assure quality of subsequent signaling processing , by avoiding windowing techniques , which can introduce substantial artifacts . tables of numbers for each standard frequency employed in the device 200 and for other frequencies in use or intended use in the device 200 are available , and are programmed into the algorithm once the user selects the test frequencies . should a combination of frequencies be required for which no common integer number can be found to fit in a practical size frame , the frame size is adjusted to j dp and the frame is windowed prior to fourier transformation , but this method is used only in extreme cases since in normal operation , the required frequencies are available . the data from a single frame is passed to a point discrete fourier transform ( dft ) block 284 which calculates the signal &# 39 ; s magnitude and phase content , but only at frequencies of interest , including f 1 , f 2 , and f dp to determine a noise floor . windowing is induced prior to processing the dft to reduce edge effects , although windowing induces energy at other bands . the block 284 is used only for temporary calculations , and the windowed data is not reused again . the output of block 284 is the magnitude and phase of primary signals at f 1 and f 2 and the noise floor figure of time at f dp . the output of block 284 forms an input to frame rejection block 286 and to an on - line calibration calculation block 288 . with the information on the magnitudes at various frequencies , a noise calculation algorithm is employed at and / or around f dp to determine the noise floor . the magnitude of the noise floor and frequency content are used against a set of predetermined conditions i . e . a comparison against an empirically derived table contained in the processor 240 , to determine the outcome of the frame . that outcome has three distinct possibilities . first , if the noise magnitude and frame content exceed a multi - threshold condition at measured frequency bands , the new frame is rejected . second , if the noise magnitudes fall between a set of reject thresholds and a set of accept thresholds , the data in the frame is disregarded , but the noise information is kept to update the noise level average . third , if the noise magnitudes are below the accept thresholds , the frame is kept and passed on for further processing and the noise magnitudes are averaged together with the noise average of the previous frame . this information is used to update thresholds , such that the system adapts to environmental conditions . when the information about magnitudes of primary tones at f 1 and f 2 , and the noise floor information at and / or around f dp , an online calibration of the level of magnitudes takes place . several actions occur in the calibration block 288 . first , if the noise floor is large when no primary tones are present , the frequency of the primaries is adjusted within predetermined limits . a new f dp is calculated , and the noise content of frequency bins at and around f dp is checked again . this process is repeated until a stable , low noise floor is established . no primary tones are played through the speaker through this process . once the primaries are presented , they are stepped up to the full output amplitude , as programmed by the user and calibrated in the ear by increasing the output of the codecs 248 . no data collection from the ear has taken place yet . at this time , if the level is not reached in a user predetermined time , and at the rate of increase of the primaries , the test is aborted due to lack of fit or the low quality of fit of the probe in the ear canal . once a proper fit of the probe in the ear canal is achieved , and testing begins , data collection takes place . during the entire process of data collection , the levels of tones at f 1 and f 2 are checked to ensure that they remain within predetermined limits throughout the test . if they exceed those limits , the output is adjusted up or down to compensate until a maximum compensation limit is reached , at which time , the test is aborted and the user is notified . also , the magnitude at and / or around f dp is continuously monitored to assure low noise floor , and if necessary , the frequency of the primary tones are adjusted on - line within predetermined limits to avoid the high external noise region . the change in frequencies of the primaries is minimal , and within the specified tolerances of the device 200 , and have been shown not to affect the magnitude of the tone within the ear at f dp . the block 290 is a store / copy buffer . as a frame of signal data is collected in new frame buffer 282 , a copy of it is saved by the store / copy buffer 290 for processing of the subsequent frames . the store / copy buffer 290 receives frame data from new frame buffer 282 and has a variable depth , depending the number of frames averaged together . buffer 290 provides an output to a slide buffer block 292 and an average buffer 294 . the slide buffer 292 operates with the stored previous frames , which are slid by a predetermined amount and the empty spaces padded with zeros for subsequent processing in the average buffer 294 . in the average buffer 294 , the frames are averaged together to reduce the uncorrelated noise present . theoretically , the noise is reduced by a factor of one over the square root of the number of averaged frames , i . e . : the frames are averaged in a linear fashion , sample by sample and a new frame is created at the end of the averaging operation . the advantage of this method is that the data is essentially correlated against a slid copy of itself , slid far enough away to avoid averaging the same information content . this provides either a substantial reduction in uncorrelated noise energy for the same amount of sampling time or a substantial reduction in sampling time to obtain the equivalent noise reduction when compared to standard linear averaging . the minimum limit to the sliding of the data , and to the reuse of old data frame is the autocorrelation function of the data in the frame , which can be predetermined or calculated on - line . this method is equivalent to taking much smaller frames and averaging them together . however , for the purposes of the subsequent fourier transformations and filtering taking place , the frame size is required to be large ( i . e ., 960 samples at 48 kilohertz sampling rate ), to obtain several full cycles of each of the tones at f 1 , f 2 , and f dp . the problem with taking a large number of very small frames is that the fourier transforms or other signal processing methods require several cycles of data for proper operation . the method of the present invention outperforms standard linear averaging of large frames because of the reduction in time as well as providing proper operation of the fourier transforms . the final average buffer 296 obtains the averaged data from the average buffer 294 , and collects it in a buffer that is used for subsequent processing and signal statistics . the output of the final average buffer 296 is digitally filtered in filter 298 which removes any remaining high or low frequency components not required for final data presentation . the averaged and filtered data is converted to frequency domain , in the embodiment illustrated , by using a discrete fourier transform at block 300 , and the resulting data then is ready for presentation to an operator as indicated at block 302 . as will be appreciated by those skilled in the art , other signal processing methods are available to convert data , and those other methods are compatible with the device 200 . in further alternate embodiments of the present invention 10 , utilized as vibration detectors or alcohol intoximeters , the input / output interface 52 is replaced with , or coupled to , one or more suitable electrical signal sensors configured to measure signals representative of the desired test material , for example , a vibration waveform or an electrical signal representative of breath alcohol content . in view of the above , it will be seen that the several objects of the invention are achieved and other advantageous results are obtained . as various changes could be made in the above constructions without departing from the scope of the invention , it is intended that all matter contained in the above description or shown in the accompanying drawings shall be interpreted as illustrative and not in a limiting sense .

a portable hand - held electrical testing device including a processor housed within an enclosure . the processor is configured to operate on commands by a user to process sub - microvolt electrical signals received through an input / output interface . the input / output interface includes a capacitive coupled amplifier with adjustable gain settings . onboard memory linked to the processor stores processing data and instructions . a display device is mounted to said enclosure and is operatively connected to the processor to display processing results in real time .

fig1 is a top plan view showing a mechanical hand or arm 20 which a child may strap on to his hand or wrist . the mechanical hand includes a control compartment and cover 22 with a hand or palm section 24 having four articulated fingers 26 and thumb 28 extending therefrom . the thumb might be either articulated or a fixed appendage . the control compartment 22 includes controls which are manipulated by a child &# 39 ; s fingers and thumb ( if an articulated thumb is provided ) for clinching the mechanical hand into a fist , or for opening the mechanical hand , or for moving the hand to assume any suitable position between a fist and an open hand . the child may also control the special effects box 39 from within the control compartment 22 . the control compartment 22 has a cover 30 ( fig2 ) hinged to the mechanical palm section 24 . a control panel 32 extends from the palm section 24 to provide a means which the child may grip and control the mechanical hand . the child places his hand and fingers on top and his thumb under the control panel 32 and then closes the cover 30 over his hand and wrist . two straps 34 , 36 ( fig5 ) are wrapped around the child &# 39 ; s wrist or forearm and then joined together in any suitable manner . while any suitable fastener may be provided to hold together the joined straps 34 , 36 , a hook and loop fastener , such as that sold under the trademark &# 34 ; velcro &# 34 ;, may be used . the child &# 39 ; s fingertips fit into rings or triggers 37 ( fig7 ) formed on slides inside the control compartment 22 . by flexing his own individual fingers and thumb , he may control the individual fingers and thumb ( if articulated ) of the mechanical hand . thus , by making his own hand into a fist , the child causes the mechanical hand to also make a fist ( fig1 ). by extending his own fingers to make an open hand , the mechanical hand also extends its fingers ( fig1 ) to become an open hand . any of a number of special effects or weapons or other add - ons may be in a box 39 which may clip onto the mechanical hand ( fig2 ). more particularly , the palm section 24 , includes not only mechanisms to articulate the fingers , but also connectors to receive the special effects box 39 . in this particular example , the special effects box 39 includes four claws 40 which may be extended or retracted by the child . a connector - actuator 41 depends from the bottom of the box 39 . this connector may be pressed into mating connectors formed on the back of the palm section 24 . once clipped on , four claws 40 operate responsive to the child &# 39 ; s manipulation of at least one of his own fingertips . in its simplest form , the claws 40 extend simultaneously with a clinching of the fist . in a more sophisticated form , the child may be given a separate control in the control compartment for the claws . many other special effects boxes may be provided for alternative attachment to the hand . for example , a &# 34 ; laser beam &# 34 ; in box 39 may shoot the enemy , at the child &# 39 ; s command . a sound device may be added to provide audible &# 34 ; attack &# 34 ; sounds or digitized voice commands . still other special effects may be provided . each of the thumb 28 and fingers 26 ( fig6 - 8 ) include a plurality of segments 42 , 43 , 44 which are hinged together in any suitable manner . more particularly , in a preferred embodiment , ( fig8 a ) the entire finger may be made in a low cost way as a single , integral , molded part , if desired . there are three segments 42 , 43 , 44 which correspond to the three segments of a human finger . a strip of plastic 45 connects each of the finger segments to its neighboring segment , preferably near the top thereof . each of these strips 45 may flex and act as a hinge which may bend as shown in fig6 . or the strips 45 may be straight so that the finger is straight , as shown in fig7 . the plastic memory of the strips 45 is such that the finger is normally straight , unless pulled in by a strap 58 . in another embodiment ( fig8 b ), each of the segments 42a , 43a , 44a has projecting parts 46 on one end which snap over mating parts on the other end . preferably , the segments 42 - 44 are molded plastic parts with a shape which simply snaps together , with no need for a hinge pin . however , it should be understood that the segments may also be held together by a hinge pin or the like . each mechanical finger segment 42 - 44 is hollow and contains a window at each end so that a sliding member in the form of a strap - like mechanism 58 may extend throughout the finger and attach at 60 to the inside of the finger tip segment 44 . attached to the palm end of the mechanism 58 is a ring or dished trigger 62 for receiving the child &# 39 ; s fingertip . trigger 62 is part of a sliding assembly . when the child &# 39 ; s fingertip pulls trigger 62 in direction a ( fig6 ), the mechanical finger curls . when the child pushes trigger 62 in the opposite direction , the finger straightens . as seen in fig9 three triggers 62 , 64 , 66 are positioned side by side within a box 68 formed in the control panel 32 . the inside dimensions of box 68 define how far forward or backward the triggers 62 - 66 may slide in order to open the mechanical hand or to clinch it in a fist . in one embodiment , the trigger 62 controls the index finger 70 of the mechanical hand . the trigger 64 controls the middle finger 72 of the mechanical hand . the trigger 66 controls the mechanical ring and little fingers , 74 , 76 . if it is articulated , the mechanical thumb 28 may be controlled by the child &# 39 ; s thumb in a similar manner . the operation of the inventive mechanical hand is shown in fig9 - 11 . the child grips the control panel 32 by placing his fingertips in the rings or dished areas of triggers 62 - 66 and his thumb under the control panel 32 . while the child is so holding the control panel 32 , cover 30 is closed and the straps 34 , 36 ( fig5 ) are secured around the child &# 39 ; s wrist or forearm . when the child closes his hand , his fingertips and thumb pull the rings or dished triggers while he closes his own fist to , in turn , close the fingers and thumb of the mechanical hand ( fig1 ). the child &# 39 ; s fingertips pull the dished triggers . when the child &# 39 ; s fingertips and push the rings or dished triggers , the mechanical fingers straighten . in one embodiment , the trigger 62 controls both the forefinger 70 and also the extension and retraction of the claws 40 in special effects box 39 . if box 39 ( fig7 ) contains an electrical device such as a laser gun or a sounding device , a push button may be located in or under the control panel 32 to be activated by a finger or thumb . in greater detail , the special effects control box 39 includes a lever arm 77 which is pivoted at 78 . the bottom 80 of the lever arm fits into a window 82 in the strap like mechanism 58 . the opposite end of lever arm 77 includes a lost motion linkage 84 which enables the lever arm to engage a pin 86 on a slide 88 . as the lever arm 77 swings back and forth , the slide 88 moves back and forth in direction e , d . this causes the claws 40 to move out to an extended position ( shown by dashed lines ) or to move back ( shown by solid lines ), in response to a movement of the trigger 62 . those who are skilled in the art will readily perceive how to modify the invention . therefore , the appended claims are to be construed to cover all equivalent structures which fall within the true scope and spirit of the invention .

a child &# 39 ; s toy is a mechanical hand having at least articulated fingers , and perhaps an articulated thumb , associated with a palm section . a control compartment attached to and extending from the palm section includes fingertip controls which selectively move the articulated fingers . a clip - on special effects box , also operated by fingertip control , may be added to the mechanical hand in order to provide a weapon , sound effect , laser beam , or the like . in the embodiment described herein , the weapon is a set of four claws which may extend from or retract into the box .

in the following paragraphs , the present invention will be described in detail by way of example with reference to the attached drawings . while this invention is capable of embodiment in many different forms , there is shown in the drawings and will herein be described in detail specific embodiments , with the understanding that the present disclosure is to be considered as an example of the principles of the invention and not intended to limit the invention to the specific embodiments shown and described . that is , throughout this description , the embodiments and examples shown should be considered as exemplars , rather than as limitations on the present invention . descriptions of well known components , methods and / or processing techniques are omitted so as to not unnecessarily obscure the invention . as used herein , the “ present invention ” refers to any one of the embodiments of the invention described herein , and any equivalents . furthermore , reference to various feature ( s ) of the “ present invention ” throughout this document does not mean that all claimed embodiments or methods must include the referenced feature ( s ). interactive toys are increasingly popular . interaction with most interactive toys occurs solely within the physical experience . these toys may contain lights , video displays , and speakers sufficient to provide an interactive experience with the user . user interaction may cause sounds to be emitted lights to flash or in some instances video to be displayed . some interactive toys are additionally coordinated with virtual worlds provided through the internet . these toys allow for limited interaction for a user to experience their toy online . one feature of the present invention is that it provides an interactive system that includes an interactive toy and an online virtual world . in this system a user may interact with a toy and through the interaction new functionality in “ unlocked ” in the virtual world . further , in some embodiments , interaction with the virtual world causes the interactive toy to respond in some manner . exemplary responses include video display , illumination of lights or light emitting diodes , emission of sounds or music from speakers , and the like . additionally , in some embodiments , new functionality may be imparted to the interactive toy based on interaction with the virtual world . as used herein , the terms “ toy ” and “ interactive toy ” are not limited by the appearance of the toy . for example , in some embodiments , the term “ toy may refer to a doll , a doll with a robot appearance , a plush toy such as a teddy - bear , a toy car , a key - chain , or virtually any other interactive device designed for play . one embodiment of a provided interactive toy is illustrated in fig1 . in this embodiment , interactive toy 10 , illustrated as a robot , it contains a toy body 50 . mounted on toy body 50 is display 20 . within toy body 10 is processor 30 and memory 40 . processor 30 is connected to display 20 . memory 40 contains multiple codes . in this embodiment , processor 30 is configured to control content displayed on display 20 . in an exemplary embodiment , memory 40 contains a set of processor executable instructions ( software ) that , when executed by processor 30 , cause an authentication code to be displayed on display 20 . as used herein the term “ authentication code ” refers to an alpha - numeric code unique to the toy . in some embodiments , this code may be used to authenticate the toy on a server on a network . in one embodiment , the set of processor executable instructions additionally configures the processor to periodically cause other codes to be displayed on display 20 . in further embodiments , display 20 provides interactive games that can be played independent of a network . this periodic display configuration may be on a periodic timed basis . for example , in one embodiment , memory 40 contains a set of 16 alpha - numeric codes . these codes may be displayed for a discrete time period over a number of days . on the first day code 1 may be displayed for 2 hours , and code 2 may be displayed for 3 hours . on the second day code 3 may be displayed for 6 hours followed by code 1 for 1 hour . in this manner the display may cycle through various combinations of codes in different days . in another embodiment , the periodic display may be controlled by interaction with interactive toy 10 . in this embodiment , processor 30 may track the interaction a user is having with the toy and once a threshold is reached cause a code to be displayed on display 20 . in a further embodiment , interactive toy 10 is configured with a number of games and controls . video portions of these games may displayed on display 20 . as a user interacts with the game processor 30 records points achieved by the user in game - play . once a certain point total is achieved , processor 30 causes a code to be displayed on display 20 . one feature of this embodiment is the displayed codes allow the user to interact with a “ virtual world ” through the use of a user computer . in one embodiment , the user computer is connected to a server on a network and the authentication code is entered a webpage and interactive toy 10 is authenticated on the network . in this embodiment , once the authentication code is received by the server , a query is run on a database . the results of the query are checked to determine if the authentication code is present in the database , and if so an account is created for the user and interactive toy 10 is authenticated on the network . in another embodiment , the server additionally hosts the virtual world and serves it to the user computer via the network . on the user computer , the virtual world is displayed for the user to interact with . various provided embodiments utilize communication technologies and networks . as is known in the art , networks can be classified in a number of ways . a network may be classified by the range supported by the underlying technology . for example , some network technologies are designed to provide communication across significant ranges . the public switched telephone network ( pstn ) is a network with almost global reach . significant portions of the pstn comprise a wired infrastructure . portions of the pstn comprise optical fiber media . other portions of the pstn infrastructure may include microwave or radio frequency links communicating across a wireless medium . wireless and cellular telephone networks interface to the pstn . the development of cellular networks has evolved over time . initially , cellular networks only supported voice traffic . more recently , data technologies have been enabled across these networks . one of the earlier cellular based data technologies was general packet radio service ( gprs ). gprs is packet based wireless communication service that provides data communication services in the 50 to 100 kilobit per second ( kbps ) speed range . the gprs standard is based on the global system for mobile ( gsm ) standard and compliments existing services such as circuit switched cellular phone connections and the short message service ( sms ). as used herein , the terms circuit switched and packet switched refer to types of network connections . circuit switched connections are typically dedicated connections between users during the entirety of the communication . packet switched refers to a connection in which packets are routed across a network based on destination and source addresses of the packets . other cellular based data technologies exist and others are currently under development , enhanced data gsm environment ( edge ) is a faster data transfer technology capable of data rates up to almost 400 kbps . like gprs , edge is based on the gsm standard and uses the gsm time division multiple access ( tdma ) frame structure . cellular or mobile based data technologies continue to evolve . the high speed downlink packet access ( hdspa ) is one of the next steps in the evolution of the universal mobile telecommunications service ( umts ). the goal of umts is to provide up to 2 megabits per second ( mbps ) to the user which will allow the transmission of text , video , voice multi - media and other services . other third generation ( 3g ) cellular efforts are additionally based on the gsm architecture . the cdma2000 based data services include evolution data optimized ( ev - do ) and 1x - evdo may additionally be employed to practice aspects of the present invention . ( cdma2000 is a registered trademark of the telecommunications industry association ). other types of data networks , may or may not interface with voice networks . these networks may have essentially global ranges , such as the internet . these networks may include wireless metropolitan area network technologies such as the technology currently referred to as wimax . metropolitan area networks may include wired or optical media as well as a wireless medium . local area networks may be wired , typically employing twisted - pair wired media , or wireless . wireless local area networks ( wlans ) may include networks employing wireless technologies like direct sequence spread spectrum ( dsss ) or orthogonal frequency division multiplexing ( ofdm ). these two wireless technologies are currently the basis for well known wlan technologies commonly referred to as wifi or 802 . 11a , b , g , and n . wireless networks may additionally be classified as wireless personal area networks ( wpan ). typically in wpan technologies the data rates can be quite significant but the ranges may be limited to under 20 meters . one example of wpan technology includes a frequency hopping spread spectrum technology , such as the technology currently known as bluetooth ( bluetooth is a registered trademark of the bluetooth special interest group ). other wpan technologies that are currently under development include ultra - wideband , which currently is being implemented as an impulse technology , a dss technology , and a frequency hopping ofdm technology . some provided embodiment embodiments employ short range communication technologies . these low range technologies may include the wpan technologies mentioned above or may comprise other low data rate low range communication transceivers . in another embodiment , illustrated in fig2 interactive toy 10 contains a communications transceiver 60 and communications port 70 . in this embodiment , interactive toy 10 may communicate codes directly to a users computer via a communications medium . various embodiment may additionally include a number of light emitting diodes and speakers ( not shown ) that assist in providing an interactive experience to a user . in various embodiments communications transceiver may be a wireless transceivers , a universal serial bus ( usb ) transceiver , a firewire transceiver , a rfid transceiver , a bluetooth transceiver , a wifi transceiver , an ultra - wideband transceiver , a modem transceiver , a wireless local area network transceiver , a wireless personal area network transceiver , a direct sequence spread spectrum transceiver , an orthogonal frequency division multiplexing transceiver , or a frequency hopping transceiver . further communications port 70 in various embodiments may be a wireless communication port comprising an antenna , a wired media communication port , a usb port , a firewire port , and a twisted pair wired media port . as is known in the art usb ports allow connection of devices and communication through the use of universal serial bus transceivers . usb is an alternative to peripheral component interconnect ( pci ) connectivity . further , firewire refers to an additional connection port and transceiver . one feature of embodiments containing communications capabilities , is that interaction within the virtual world may be communicated to interactive toy 10 via communication port 70 and communications transceiver 60 . for example , a user may achieve a point total or other interaction with a virtual copy of the interactive toy 10 . in some embodiments , this achievement causes the server to send a communication to the user &# 39 ; s computer . the user computer then communicates with interactive toy 10 causing an event to occur . exemplary events include but are not limited to illumination of leds 120 , emission of music or other sounds from the speaker ( not shown ), display of images or video on display 20 , display of codes on display 20 , or other such action . an exemplary embodiment of an interactive play system is illustrated in fig3 . the system includes an interactive toy 10 having a display 20 . other features , described above , are omitted from fig3 for convenience . in this embodiment , interactive toy 10 is configured to communicate with user computer 80 via a wireless communication medium . user computer 80 includes a display 20 , a processor 30 , memory 40 , and communications transceiver 60 . in some embodiments , user computer may comprise additional communications transceivers 60 ( one shown for convenience ). user computer 70 additionally includes communications ports 70 . in fig3 one communications port 70 is illustrated with an antenna for wireless connectivity to interactive toy 10 and another communications port 70 is illustrated providing connection to network 90 . as is known in the art , connection to network 90 may be through a wireless or wired media . user computer 80 is additionally illustrated with keyboard 250 . additional components are omitted for convenience . one feature of an embodiment of interactive play system is that codes may be displayed on display 20 on interactive toy 10 . in embodiments where interactive toy 10 does not include a communications transceiver 60 a user may enter the codes through keyboard 250 . in other embodiments , codes may be communicated directly from interactive toy 10 to user computer 80 through communications transceivers 60 and communications ports 70 . another feature of various embodiments of the interactive play system is that interactive toy 10 may be authenticated on server 100 . once authenticated , user computer 80 may display a virtual world environment on display 20 allowing a user to interact with the virtual world . in various embodiments , the virtual world contains a virtual toy 130 with an appearance of interactive toy 10 . one aspect of an embodiment containing a virtual toy 130 is that user interactions with virtual toy 130 may cause events to happen on interactive toy 10 . for example , a user may play a game in the virtual world that involves interactive toy 10 . once an event occurs in the virtual world , lights or leds 120 on interactive toy 10 may light , sounds or songs may be played on interactive toy 10 , or images or video displayed on display 20 on interactive toy 10 . as described above , network 90 may be any form of network 90 , but in an exemplary embodiment is the internet . server 100 is additionally connected to network 90 through communications port 70 and is configured to communicate with user computer 80 via communications transceiver 60 . server 100 contains a processor 30 , memory 40 , and in some embodiments storage media 140 . server 100 additionally includes database 110 . one feature provided by embodiments of the interactive play system is that interactive toy 10 is authenticated on server 100 and a user is provided a virtual world on user computer 80 . as the user interacts with the virtual world , server 100 stores data related to interactive toy 10 , the user , and his / her activities in the virtual world . as a user achieves certain objectives in the virtual world , accrued points and other data , such as a set of codes for each interactive toy 10 , are stored on storage medium 140 . once point totals exceed a threshold value , or other objectives are met , server 100 communicates the accomplishment back to user computer 80 which , in some embodiments , are communicated to interactive toy 10 . further , as described above , interactive toy 10 periodically displays codes on display 10 . these codes , once authenticated on server 100 may provide additional features to the virtual world . for example , a user &# 39 ; s virtual toy 130 may be enabled with additional functionality , secret rooms or spaces may be opened for virtual toy 130 to explore , additional virtual point totals may be assigned , access to additional virtual worlds , access to additional virtual objects , access to additional virtual cash , and access to additional virtual games to name a few . in some embodiments , each code displayed is unique to an add - on functionality . further , in some embodiments are displayed on a time periodic basis , they may expire if not authenticated within a predetermined time period . in various embodiments , the virtual world hosted and served by server 100 , is displayed on user computer 80 through a web - browser interface . in one such embodiment the web - browser interface contains a plug - in that allows animation and control . many browser plug - ins are well known in the art and may be used to practice the current invention . an exemplary plug - in is a flash control plug - in . the browser - interface and plug - in are configured to provide a graphical and in some instances audible experience to a user . an exemplary provided method is illustrated in fig4 . flow begins in block 150 where an authentication code is displayed on the display 20 of an interactive toy 10 . in block 160 the toy is authenticated on a server 100 on network 90 . flow continues to block 170 where server 100 provides a virtual world to a user computer 80 . in block 180 a code is selected from a set of codes contained in memory 40 of interactive toy 10 . in this embodiment , the selected code is not displayed on display 20 of interactive toy 10 . this code is validated in block 190 . flow continues to block 200 where additional functionality is provided by server 100 . in some embodiments the codes are entered into user computer 80 through keyboard 250 . in other embodiments codes are communicated from interactive toy 10 to user computer 80 through the use of communications transceivers 60 and communications ports 70 . user computer 80 then communicates the codes to server 100 across network 90 . authentication or validation of a code at server 100 may comprise executing a query on database 110 to determine if the code is stored in database 110 . as discussed above , the additional functionality provided in block 200 may be additional functionality within the virtual world or in some embodiments may be additional functionality communicated to interactive toy 10 . fig5 illustrated an additional embodiment of a provided method . like the above embodiment flow begins at block 150 where an authentication code is displayed on the display 20 of an interactive toy 10 . in block 160 the toy is authenticated on a server 100 on network 90 . flow continues to block 170 where server 100 provides a virtual world to a user computer 80 . in block 180 a code is selected from a set of codes contained in memory 40 of interactive toy 10 . flow continues to block 210 where the selected code is displayed on display 20 of interactive toy 10 . flow continues to block 200 where additional functionality is provided by server 100 . the remainder of the method is the same as the method illustrated in fig4 . fig6 illustrates another embodiment of a provided method . this embodiment functions the same as fig . for with the exception of conditional block 220 . in this block interactive toy 10 determines is an appropriate condition is met before selecting and displaying a code . until the condition is met the method remains in condition block 220 . once the condition is met flow continues back to block 180 where a new code is selected . as discussed above , in various embodiments codes are released based meeting conditions . one condition is passage of time . in this embodiment , codes are selected and displayed on a time periodic basis . another condition is a level of interaction with the interactive toy 10 . a further condition is the achievement of a point total in a game . fig7 - 12 illustrate various embodiments of interactive toy 10 . embodiments of interactive toy 10 ( a ) through 10 ( e ), shown in fig7 illustrate exemplary functionality of displays 20 ( a ) through 20 ( e ). in some embodiments like display 20 ( c ) may be used to display messages or advertisements . in other embodiments , illustrated in interactive toy 10 ( d ), display 20 ( d ) may be used to display video or still imagery , in further embodiments , illustrated in interactive toy 10 ( e ), display 20 ( e ) is used to display a video game . in another embodiment of interactive toy 10 ( b ), display 20 ( b ) is used to display a mood . fig8 illustrates another embodiment of interactive toy 10 , configured with display 20 and communications connector 350 . as is known in the art , this embodiment contains communications transceivers and ports capable of communications on communications connector 350 . in this embodiment , communications connector 350 may be any wired media connector , such as a usb , firewire , twisted pair , s - video , or coaxial connector to name a few examples . another embodiment of interactive toy 10 is illustrated in fig9 . in this embodiment interactive toy 10 is equipped with display 20 , capable of displaying alpha - numeric codes , led 120 , speaker 430 , communications port 70 and communications connector 350 . one feature of this embodiment is that it is able to communicate codes directly to user computer 80 , and receive communications from user computer 80 , through communications port 80 and communications connector 350 . an alternate embodiment is illustrated in fig1 . a primary distinction between this and the embodiment illustrated in fig9 , is this embodiment is not equipped with the ability to communicate with user computer 80 . a feature of this embodiment is that codes displayed on display 20 must be manually entered into user computer 80 with keyboard 250 . fig1 illustrated a further embodiment of interactive toy 10 . in this embodiment , toy body 50 is fabric and interactive toy 10 is a stuffed “ plush ” toy . fig1 further illustrates multiple communications ports 70 , which require multiple communications transceivers 60 . one communications transceiver being suitable for wired media communication over communications connector 350 , another being suitable for communication over a wireless medium . fig1 further illustrates embodiments of interactive toy 10 that include controls 440 . controls 440 , depicted here as on / off controls , in some embodiments include controls sufficient to control a video game displayed on display 20 . a further embodiment of interactive toy 10 is illustrated in fig1 . in this embodiment interactive toy 10 is a key chain having a toy body 50 , display 20 and controls 440 . as stated above , in embodiments where interactive toy 10 is capable of displaying a video game on display 20 , further controls 440 are present allowing a user to interact with the video game . one feature of this embodiment is that in embodiments where codes are displayed on a time periodic basis , a user is able to carry interactive toy 10 with them throughout the day and record the code when displayed ensuring they do not miss the display . at the heart of the interactive system provided by one embodiment is the user computer 80 . certain provided architecture and functionality is illustrated in fig1 - 15 . fig1 illustrates an embodiment of user computer 80 . in this embodiment , user computer 80 is configured with browser environment 260 and driver 290 . the virtual world hosted by server 100 is provided to a user through browser environment 260 . in one embodiment , browser environment contains a web - browser with a software plug - in 270 . software plug - in 270 is capable of visually displaying the virtual world on display 20 of user computer 80 . many software plug - ins are known in the art and may be used to practice the current invention . an exemplary plug - in is a “ flash ” plug - in well known in the art . in some embodiments , software plug - in 270 is capable of writing a file 280 through a write command . in these embodiments , the contents of file 280 are related to activities within the virtual world . many file formats are known in the art and may be used to practice various embodiments the invention . an exemplary file is commonly known as a “ cookie ”. user computer 80 , in embodiments where interactive toy 10 is capable of communication , is configured with driver 290 . driver 290 contains interpreter 300 which is configured to retrieve file 280 with a pull command , interpret the contents of file 280 and send action signals to interactive toy 10 . the specific actions are described above . further , in embodiments where interactive toy , can communicate with user computer 80 , driver 290 may be configured to receive codes and other data from interactive toy 10 and communicate it to server 100 across network 90 . operational flow of an exemplary driver 290 is illustrated in fig1 . in block 310 driver 290 issues a pull command to retrieve file 280 . flow continues to conditional block 320 where driver 290 determines if file 280 exists . if file 280 does not exist , flow returns to block 310 and periodically repeated . if it is determined in conditional block 320 that file 280 exists , flow continues to block 330 and file 280 is retrieved and interpreted by interpreter 300 . flow continues to conditional block 340 where the validity of file 280 is determined . if the file is not valid , flow continues back to block 310 . if conditional block validates file 280 , flow continues to block 350 where a signal is sent to interactive toy 10 . in block 360 interactive toy 10 responds to the signal by performing an action . flow continues to block 370 where file 280 is deleted . operational flow of an exemplary browser environment 260 is illustrated in fig1 . server 100 provides virtual world to browser environment 260 . flow begins in block 390 where a user is playing on the provided virtual world . flow continues to conditional block 400 where it is determined if the user accomplishes a predetermined goal within the virtual world . if the user has not achieved the goal flow remains within conditional block 400 until the goal is achieved . once the goal is achieved , flow continues to conditional block 410 where it is determined if the achieved goal is one that should cause an action by interactive toy 10 . if it is not flow continues back to conditional block 400 and waits for the next accomplishment . if conditional block 410 determines that the accomplished goal is one where interactive toy 10 action is appropriate , flow continues to block 420 where software plug - in 270 writes file 280 . thus , it is seen that a system , methods and apparatus for interactive play are provided . one skilled in the art will appreciate that the present invention can be practiced by other than the above - described embodiments , which are presented in this description for purposes of illustration and not of limitation . the specification and drawings are not intended to limit the exclusionary scope of this patent document . it is noted that various equivalents for the particular embodiments discussed in this description may practice the invention as well . that is , while the present invention has been described in conjunction with specific embodiments , it is evident that many alternatives , modifications , permutations and variations will become apparent to those of ordinary skill in the art in light of the foregoing description . accordingly , it is intended that the present invention embrace all such alternatives , modifications and variations as fall within the scope of the appended claims . the fact that a product , process or method exhibits differences from one or more of the above - described exemplary embodiments does not mean that the product or process is outside the scope ( literal scope and / or other legally - recognized scope ) of the following claims .

a system , method and apparatus for interactive play is provided . in an exemplary embodiment , an interactive toy is provided . the interactive toy is configured to store a plurality of codes and display codes on a display on the toy . the codes can authenticate the toy and unlock functionality within a virtual world . in one embodiment , the interactive toy is configured to communicate with a user computer . another embodiment provides methods for interactive play with an interactive toy and a virtual world . a further embodiment provides a computing device configured to participate in the interactive play system . this abstract is provided for the purpose of complying with the abstract requirement rules that allow a reader to quickly ascertain the subject matter of the disclosure contained herein . this abstract is submitted with understanding that it will not be used to interpret or to limit the scope or the meaning of the claims .

a soybean cultivar needs to be highly homogeneous , homozygous and reproducible to be useful as a commercial cultivar . there are many analytical methods available to determine the homozygotic and phenotypic stability of these cultivars . the oldest and most traditional method of analysis is the observation of phenotypic traits . the data is usually collected in field experiments over the life of the soybean plants to be examined . phenotypic characteristics most often observed are for traits associated with seed yield , lodging resistance , disease resistance , emergence , maturity , plant height , shattering , flower color , pubescence color , pod color and hilum color . in addition to phenotypic observations , the genotype of a plant can also be examined . there are many laboratory - based techniques available for the analysis , comparison and characterization of plant genotype ; among these are isozyme electrophoresis , restriction fragment length polymorphisms ( rflps ), randomly amplified polymorphic dnas ( rapds ), arbitrarily primed polymerase chain reaction ( ap - pcr ), dna amplification fingerprinting ( daf ), sequence characterized amplified regions ( scars ), amplified fragment length polymorphisms ( aflps ), and simple sequence repeats ( ssrs ) which are also referred to as microsatellites . the cultivar of the invention has shown uniformity and stability for all traits , as described in the following cultivar description information . it has been self - pollinated a sufficient number of generations , with careful attention to uniformity of plant type to ensure homozygosity and phenotypic stability . the line has been increased with continued observation for uniformity . no variant traits have been observed or are expected in s56 - d7 . soybean cultivar s56 - d7 , being substantially homozygous , can be reproduced by planting seeds of the line , growing the resulting soybean plants under self - pollinating or sib - pollinating conditions , and harvesting the resulting seed , using techniques familiar to the agricultural arts . publications useful as references in interpreting the breeding history and data presented below include : caldwell , b . e . ed . 1973 . “ soybeans : improvement , production , and uses ” amer . soc . agron . monograph no . 16 ; buttery , b . r ., and r . i . buzzell 1968 . “ peroxidase activity in seed of soybean varieties ” crop sci . 8 : 722 - 725 ; hymowitz , t . 1973 . “ electrophoretic analysis of sbti - a2 in the usda soybean germplasm collection ” crop sci ., 13 : 420 - 421 ; payne r . c ., and l . f . morris , 1976 . “ differentiation of soybean varieties by seedling pigmentation patterns ” j . seed . technol . 1 : 1 - 19 . the disclosures of which are each incorporated by reference in their entirety . s56 - d7 was selected from an f 4 plant from the cross x9757 / b530142r . x9757 is a breeding line developed by syngenta seeds , inc . from the cross hutcheson / s57 - 11 . b530142r is a breeding line developed by syngenta seeds , inc . from the cross s59 - 60 / 40 - 3 - 2 . s57 - 11 and s59 - 60 are commercial cultivars developed by syngenta seeds , inc . hutcheson is a public cultivar developed and released by the virginia agricultural experiment station . 40 - 3 - 2 is a line developed by monsanto , which carries their patented gene conferring tolerance to roundup herbicide . the cross between x9757 and b530142r was made at kekaha , kauai , hi in february 1996 . the f 1 generation was grown at the at bay , ar in the summer of 1996 , the f 2 and f 3 at kekaha in the winter of 1996 - 97 , and the f 4 at bay in the summer of 1997 . all four generations were sprayed with roundup to eliminate any susceptible plants . single f 4 plants were harvested and threshed individually . each single - plant derived line was tested in a second year retest in 1999 and then in advanced trials in the southern usa in 2000 and 2001 . seed was purified and increased in puerto rico in the winter of 1999 - 2000 and at bay in the summer of 2000 . s56 - d7 was tested in the greenhouse at bay for resistance to soybean cyst nematode in 1998 , 1999 , 2000 , and 2001 and found to be resistant to races 1 , 3 , 9 , and 14 . it was also screened for resistance to southern stem canker in the greenhouse at bay in 2000 , 2001 , and 2002 and for resistance to frogeye leaf spot in the field at bay in 2000 and 2001 . s56 - d7 was found to be resistant to both diseases . flower color purple pubescence color tawny pod color tan hilum color black leaf shape ovate stem termination determinate plant type intermediate hypocotyl color light purple maturity group 5 relative maturity 5 . 6 soybean cyst nematode resistant ( r 1 , 3 , 9 , 14 ) southern stem canker resistant frogeye leaf spot resistant hypocotyl length long the invention is further directed to methods for producing a soybean plant by crossing a first parent soybean plant with a second parent soybean plant , wherein the first or second soybean plant is the soybean plant from the soybean line s56 - d7 . further , both first and second parent soybean plants may be from the soybean cultivar s56 - d7 . therefore , any methods using the soybean cultivar s56 - d7 are part of this invention : selfing , backcrosses , hybrid breeding and crosses to populations . all plants produced using soybean cultivar s56 - d7 as a parent are within the scope of the invention . the invention also encompasses plants of cultivar s56 - d7 and parts thereof further comprising one or more specific , single gene transferred traits . such traits are introgressed into cultivar s56 - d7 from another soybean cultivar or are directly transformed into cultivar s56 - d7 . preferably , one or more new traits are transferred to cultivar s56 - d7 , or , alternatively , one or more traits of cultivar s56 - d7 are altered or substituted . the introgression of the trait ( s ) into cultivar s56 - d7 is for example achieved by recurrent selection breeding , for example by backcrossing . the goal of a backcross protocol is to alter or substitute a single trait or characteristic in the original inbred . in one embodiment of the present invention , cultivar s56 - d7 ( the recurrent parent ) is first crossed to a donor inbred ( the non - recurrent parent ) that carries the appropriate gene ( s ) for the trait ( s ) in question . the progeny of this cross is then mated back to the recurrent parent followed by selection in the resultant progeny for the desired trait ( s ) to be transferred from the non - recurrent parent . after three , preferably four , more preferably five or more generations of backcrosses with the recurrent parent with selection for the desired trait ( s ), the progeny will be heterozygous for loci controlling the trait ( s ) being transferred , but will be like the recurrent parent for most or almost all other genes , i . e ., will be like the recurrent parent for essentially all of the recurrent parent &# 39 ; s physiological and morphological characteristics . ( see , for example , poehlman & amp ; sleper ( 1995 ) breeding field crops , 4th ed ., 172 - 175 ; fehr ( 1987 ) principles of cultivar development , vol . 1 : theory and technique , 360 - 376 ). the laboratory - based techniques described above , in particular rflp and ssr , can be used in such backcrosses to identify the progenies having the highest degree of genetic identity with the recurrent parent . this permits one to accelerate the production of soybean cultivars having at least 90 %, preferably at least 95 %, more preferably at least 99 % genetic identity with the recurrent parent , yet more preferably genetically identical to the recurrent parent , and further comprising the trait ( s ) introgressed from the donor patent . such determination of genetic identity can be based on molecular markers used in the laboratory - based techniques described above . the last backcross generation is then selfed to give pure breeding progeny for the gene ( s ) being transferred . the resulting plants have essentially all of the morphological and physiological characteristics of cultivar s56 - d7 , in addition to the single gene trait ( s ) transferred to the inbred . the exact backcrossing protocol will depend on the trait being altered to determine an appropriate testing protocol . although backcrossing methods are simplified when the trait being transferred is a dominant allele , a recessive allele may also be transferred . in this instance it may be necessary to introduce a test of the progeny to determine if the desired trait has been successfully transferred . the cultivar of the invention can also be used for transformation where exogenous genes are introduced and expressed by the cultivar of the invention . genetic variants created either through traditional breeding methods using cultivar s56 - d7 or through transformation of cultivar s56 - d7 by any of a number of protocols known to those of skill in the art are intended to be within the scope of this invention ( see e . g . trick et al . ( 1997 ) recent advances in soybean transformation , in plant tissue culture and biotechnology , 3 : 9 - 26 , incorporated herein by reference ). production of a genetically modified plant tissue by transformation combines teachings of the present disclosure with a variety of techniques and expedients known in the art . in most instances alternate expedients exist for each stage of the overall process . the choice of expedients depends on the variables such as the plasmid vector system chosen for the cloning and introduction of the desired recombinant dna molecule , as well as the particular structural gene , promoter elements and upstream elements used . persons skilled in the art are able to select and use appropriate alternatives to achieve functionality . culture conditions for expressing desired structural genes and cultured cells are known in the art . also as known in the art , soybeans are transformable and regenerable such that whole plants containing and expressing desired genes under regulatory control may be obtained . general descriptions of plant expression vectors and reporter genes and transformation protocols can be found in gruber , et al ., “ vectors for plant transformation , in methods in plant molecular biology & amp ; biotechnology ” in glich et al ., ( eds . pp . 89 - 119 , crc press , 1993 ). moreover gus expression vectors and gus gene cassettes are available from clone tech laboratories , inc ., palo alto , calif . while luciferase expression vectors and luciferase gene cassettes are available from pro mega corp . ( madison , wis .). general methods of culturing plant tissues are provided for example by maki et al . “ procedures for introducing foreign dna into plants ” in methods in plant molecular biology & amp ; biotechnology , glich et al . ( eds . pp . 67 - 88 crc press , 1993 ); and by phillips et al . “ cell - tissue culture and in - vitro manipulation ” in soybean & amp ; soybean improvement , 3rd edition sprague et al . ( eds . pp . 345 - 387 ) american society of agronomy inc . et al . 1988 . methods of introducing desired recombinant dna molecule into plant tissue include the direct infection or co - cultivation of plant cells with agrobacterium tumefaciens , horsch et al ., science , 227 : 1229 ( 1985 ). descriptions of agrobacterium vector systems and methods for agrobacterium - mediated gene transfer provided by gruber , et al . supra . other useful methods include but are not limited to expression vectors introduced into plant tissues using a direct gene transfer method such as microprojectile - mediated delivery , dna injection , electroporation and the like . more preferably expression vectors are introduced into plant tissues using the biolistic microprojectile delivery or agrobacterium - medicated transformation . transformed plants obtained via protoplast transformation are also intended to be within the scope of this invention . many traits have been identified that are not regularly selected for in the development of a new cultivar but that can be improved e . g . by backcrossing techniques or by genetic transformation . using materials and methods well known to those persons skilled in the art , traits that are capable of being transferred , to cultivar s56 - d7 include , but are not limited to , herbicide tolerance , resistance for bacterial , fungal , or viral disease , nematode resistance , insect resistance , enhanced nutritional quality , such as oil , starch and protein content or quality , improved performance in an industrial process , altered reproductive capability , such as male sterility or male fertility , yield stability and yield enhancement . other traits transferred to cultivar s56 - d7 are for the production of commercially valuable enzymes or metabolites in plants of cultivar s56 - d7 . traits capable of being transferred to soybean cultivar s56 - d7 are naturally occurring soybean traits or transgenic traits . transgenes are directly introduced into cultivar s56 - d7 using genetic engineering and transformation techniques well known in the art , some of which are described above , or are originally introduced into a donor , non - recurrent parent using genetic engineering and transformation techniques , which are then introgressed into cultivar s56 - d7 , for example by backcrossing . a transgene typically comprises a nucleotide sequence whose expression is responsible or contributes to the trait , under the control of a promoter capable of directing the expression of the nucleotide sequence at the desired time in the desired tissue or part of the plant . constitutive , tissue - specific or inducible promoters preferably are used . the transgene may also comprise other regulatory elements such as for example translation enhancers or termination signals . in one embodiment of the present invention , the transgene nucleotide sequence in the cultivar s56 - d7 includes a coding sequence that is transcribed and translated into a protein . in another embodiment of the invention , the nucleotide sequence encodes an antisense rna or a sense rna that is not translated or only partially translated . where more than one trait are introgressed into cultivar s56 - d7 , it is preferred that the specific genes are all located at the same genomic locus in the donor , non - recurrent parent , preferably , in the case of transgenes , as part of a single dna construct integrated into the donor &# 39 ; s genome . alternatively , if the genes are located at different genomic loci in the donor , non - recurrent parent , backcrossing allows to recover essentially all of the morphological and physiological characteristics of cultivar s56 - d7 in addition to the multiple genes in the resulting soybean cultivar . the genes responsible for a specific , single gene trait are generally inherited through the nucleus . known exceptions are , e . g . the genes for male sterility , some of which are inherited cytoplasmically , but still act as single gene traits . in a preferred embodiment , a transgene to be introgressed into cultivar s56 - d7 is integrated into the nuclear genome of the donor , non - recurrent parent or the transgene is directly transformed into the nuclear genome of cultivar s56 - d7 . in another preferred embodiment , a transgene to be introgressed into cultivar s56 - d7 is integrated into the plastid genome of the donor , non - recurrent parent or the transgene is directly transformed into the plastid genome of cultivar s56 - d7 . in a preferred embodiment , a plastid transgene comprises one gene transcribed from a single promoter or two or more genes transcribed from a single promoter . a non - exclusive list of traits or nucleotide sequences capable of being transferred into cultivar s56 - d7 , using material and methods well known to those persons skilled in the art are as follows : genetic factor ( s ) responsible for resistance to brown stem rot ( u . s . pat . no . 5 , 689 , 035 ) or resistance to cyst nematodes ( u . s . pat . no . 5 , 491 , 081 ); a transgene encoding an insecticidal protein , such as , for example , a crystal protein of bacillus thuringiensis or a vegetative insecticidal protein from bacillus cereus , such as vip3 ( see for example estruch et al . nat biotechnol ( 1997 ) 15 : 137 - 41 ; a herbicide tolerance transgene whose expression renders plants of cultivar s56 - d7 tolerant to the herbicide , for example , expression of an altered acetohydroxyacid synthase ( ahas ) enzyme confers upon plants tolerance to various imidazolinone or sulfonamide herbicides ( u . s . pat . no . 4 , 761 , 373 ). other such traits include , for example , a non - transgenic trait conferring to cultivar s56 - d7 tolerance to imidazolinones or sulfonylurea herbicides ; a transgene encoding a mutant acetolactate synthase ( als ) that render the plants resistant to inhibition by sulfonylurea herbicides ( u . s . pat . no . 5 , 013 , 659 ); a gene encoding a mutant glutamine synthetase ( gs ) resistant to inhibition by herbicides that are known to inhibit gs , e . g . phosphinothricin and methionine sulfoximine ( u . s . pat . no . 4 , 975 , 374 ); and a streptomyces bar gene encoding a phosphinothricin acetyl transferase resulting in tolerance to the herbicide phosphinothricin or glufosinate ( u . s . pat . no . 5 , 489 , 520 ). other traits capable of being transferred to the cultivar s56 - d7 of the invention include toleration to inhibition by cyclohexanedione and aryloxyphenoxypropanoic acid herbicides ( u . s . pat . no . 5 , 162 , 602 ), which is conferred by an altered acetyl coenzyme a carboxylase ( accase ); transgenic glyphosate tolerant plants , which tolerance is conferred by an altered 5 - enolpyruvyl - 3 - phosphoshikimate ( epsp ) synthase gene ; tolerance to a protoporphyrinogen oxidase inhibitor , which is achieved by expression of a tolerant protoporphyrinogen oxidase enzyme in plants ( u . s . pat . no . 5 , 767 , 373 ). in yet another embodiment of the present invention , a transgene introgressed into cultivar s56 - d7 comprises a gene conferring tolerance to a herbicide and at least another nucleotide sequence for another trait , such as for example , insect resistance or tolerance to another herbicide . direct selection may be applied where the trait acts as a dominant trait . an example of a dominant trait is herbicide tolerance . for this selection process , the progeny of the initial cross are sprayed with the herbicide prior to the backcrossing . the spraying eliminates any plant which do not have the desired herbicide tolerance characteristic , and only those plants which have the herbicide tolerance gene are used in the subsequent backcross . this process is then repeated for the additional backcross generations . this invention is also directed to methods for producing a soybean plant by crossing a first parent soybean plant with a second parent soybean plant , wherein the first or second soybean plant is the soybean plant from the line s56 - d7 . further , both first and second parent soybean plants may be from the cultivar s56 - d7 . therefore , any methods using the cultivar s56 - d7 are part of this invention : selfing , backcrosses , hybrid breeding , and crosses to 1 5 populations . any plants produced using cultivar s56 - d7 or cultivar s56 - d7 further comprising one or more specific , single gene traits as a parent are within the scope of this invention . for example , the soybean cultivar s56 - d7 or cultivar s56 - d7 further comprising one or more specific , single gene traits are used in crosses with other , different , soybean plants to produce first generation ( f 1 ) soybean hybrid seeds and plants with superior characteristics . for example , a method to produce a hybrid soybean seed comprises the steps of planting , preferably in pollinating proximity , seeds of soybean cultivar s56 - d7 or seeds of soybean cultivar s56 - d7 further comprising one or more specific , single gene traits and another soybean cultivar , cultivating the soybean plants resulting from said seeds until said plants bear flowers , emasculating the plants of either one or the other soybean cultivar , inducing cross pollination to occur between said soybean cultivars and harvesting seeds produced on said emasculated plants of the cultivar line . as used herein , the term “ plant ” includes plant cells , plant protoplasts , plant cells of tissue culture from which soybean plants can be regenerated , plant calli , plant clumps , and plant cells that are intact in plants or parts of plants , such as pollen , flowers , seeds , pods , leaves , stems , and the like . thus , another aspect of this invention is to provide for cells that upon growth and differentiation produce the cultivar s56 - d7 . further reproduction of the cultivar can occur by tissue culture and regeneration . tissue culture of various tissues of soybeans and regeneration of plants therefrom is well known and widely published . for example , reference may be had to komatsuda , t . et al ., “ genotype x sucrose interactions for somatic embryogenesis in soybean ,” crop sci . 31 : 333 - 337 ( 1991 ); stephens , p . a . et al ., “ agronomic evaluation of tissue - culture - derived soybean plants ,” theor . appl . genet . ( 1991 ) 82 : 633 - 635 ; komatsuda , t . et al ., “ maturation and germination of somatic embryos as affected by sucrose and plant growth regulators in soybeans glycine gracilis skvortz and glycine max ( l .) merr .,” plant cell , tissue and organ culture , 28 : 103 - 113 ( 1992 ); dhir , s . et al ., “ regeneration of fertile plants from protoplasts of soybean ( glycine max l . merr . ): genotypic differences in culture response ,” plant cell reports ( 1992 ) 11 : 285 - 289 ; pandey , p . et al ., “ plant regeneration from leaf and hypocotyl explants of glycine wightii ( w . and a .) verdc . var longicauda ,” japan j . breed . 42 : 1 - 5 ( 1992 ); and shetty , k ., et al ., “ stimulation of in vitro shoot organogenesis in glycine max ( merrill .) by allantoin and amides ,” plant science 81 : ( 1992 ) 245 - 251 ; as well as u . s . pat . no . 5 , 024 , 944 , issued jun . 18 , 1991 to collins et al . and u . s . pat . no . 5 , 008 , 200 , issued apr . 16 , 1991 to ranch et al . thus , another aspect of this invention is to provide cells which upon growth and differentiation produce soybean plants having all or essentially all the physiological and morphological characteristics of cultivar s56 - d7 . the disclosures , publications , and patents which are disclosed herein are all hereby incorporated herein in their entirety by reference . the seed of soybean cultivar s56 - d7 further comprising one or more specific , single gene traits , the plant produced from the seed , the hybrid soybean plant produced from the crossing of the cultivar with any other soybean plant , hybrid seed , and various parts of the hybrid soybean plant can be utilized for human food , livestock feed , and as a raw material in industry . soybean is the world &# 39 ; s leading source of vegetable oil and protein meal . the oil extracted from soybeans is used for cooking oil , margarine , and salad dressings . soybean oil is composed of saturated , monounsaturated and polyunsaturated fatty acids . it has a typical composition of 11 % palmitic , 4 % stearic , 25 % oleic , 50 % linoleic and 9 % linolenic fatty acid content (“ economic implications of modified soybean traits summary report ”, iowa soybean promotion board & amp ; american soybean association special report 92s , may 1990 . changes in fatty acid composition for improved oxidative stability and nutrition are constantly sought after . industrial uses of soybean oil which is subjected to further processing include ingredients for paints , plastics , fibers , detergents , cosmetics , and lubricants . soybean oil may be split , inter - esterified , sulfurized , epoxidized , polymerized , ethoxylated , or cleaved . designing and producing soybean oil derivatives with improved functionality , oliochemistry , is a rapidly growing field . the typical mixture of triglycerides is usually split and separated into pure fatty acids , which are then combined with petroleum - derived alcohols or acids , nitrogen , sulfonates , chlorine , or with fatty alcohols derived from fats and oils . soybean is also used as a food source for both animals and humans . soybean is widely used as a source of protein for animal feeds for poultry , swine and cattle . during processing of whole soybeans , the fibrous hull is removed and the oil is extracted . the remaining soybean meal is a combination of carbohydrates and approximately 50 % protein . for human consumption soybean meal is made into soybean flour which is processed to protein concentrates used for meat extenders or specialty pet foods . production of edible protein ingredients from soybean offers a healthy , less expensive replacement for animal protein in meats as well as dairy - type products . [ 0062 ] table 2 comparison between s56 - d7 , syngenta s58 - r3 , ag5701 , pioneer p95b53 , and syngenta s57 - a4 data summary , 2001 yield lodging height prr scn cultivar ( bu / ac ) maturity ( 1 - 9 ) ( cm ) ( 1 - 9 ) r3 r14 s56 - d7 47 . 4 oct - 10 3 . 0 87 4 . 4 r r s58 - r3 46 . 4 oct - 11 2 . 7 85 4 . 3 r r ag5701 46 . 2 oct - 12 2 . 0 86 4 . 9 — — pioneer 45 . 5 oct - 7 1 . 9 78 3 . 9 — — p95b53 s57 - a4 44 . 9 oct - 8 2 . 3 88 4 . 1 mr mr grand 45 . 5 oct - 9 2 . 4 88 4 . 5 — — mean no . of 21 7 11 15 1 — — tests lsd 2 . 6 2 0 . 7 4 0 . 9 — — ( 0 . 05 ) applicants have made a deposit of at least 2500 seeds of the cultivar of the present invention with the american type culture collection ( atcc ), manassas , va ., 20110 - 2209 u . s . a ., atcc accession number no : ______ . this deposit of cultivar s56 - d7 will be maintained in the atcc depository , which is a public depository , for a period of 30 years , or 5 years after the most recent request , or for the effective life of the patent , whichever is longer , and will be replaced if it becomes nonviable during that period . additionally , applicants have satisfied all the requirements of 37 c . f . r . §§ 1 . 801 - 1 . 809 , including providing an indication of the viability of the sample . applicants impose no restrictions on the availability of the deposited material from the atcc ; however , applicants have no authority to waive any restrictions imposed by law on the transfer of biological material or its transportation in commerce . applicants do not waive any infringement of its rights granted under this patent or under the plant variety protection act ( 7 usc 2321 et seq .). the foregoing invention has been described in detail by way of illustration and example for purposes of clarity and understanding . however , it will be obvious that certain changes and modifications such as single gene modifications and mutations , somaclonal variants , variant individuals selected from large populations of the plants of the instant inbred and the like may be practiced within the scope of the invention , as limited only by the scope of the appended claims .

a novel soybean cultivar designated s56 - d7 with high yield potential , tolerance to roundup herbicide , mid mg v maturity , and resistant to frogeye leaf spot , southern stem canker and races 1 , 3 , 9 and 14 of soybean cyst nematode , further including the plants and seeds of the cultivar s56 - d7 , methods for producing a soybean plant by crossing the cultivar s56 - d7 with itself or another soybean plant . the invention also relates to soybean cultivar s56 - d7 further comprising one or more single gene traits , and to methods of producing a soybean having such traits by transformation or mutagenesis . the invention also includes using the soybean cultivar s56 - d7 to produce other soybean cultivars , breeding lines , and progeny

referring to fig1 and 2 , a bolus is shown which contains therapeutic , additive or nutrient substances for delivery into the reticulorumen of a ruminant over a prolonged period of time . the bolus comprises a substantially cylindrical body or shell 10 having a hollow interior chamber 12 , an open end 14 , and a closed end 16 . the body 10 is made from a material or is coated with a material which is resistant to biochemical activity inside the reticulorumen . for example , body 10 may be made from a resistant epoxy , polyethylene , or polypropylene . alternatively , body 10 may be made from wood or metal and coated with a resistant epoxy , polyethylene , or polypropylene . inside the bolus , partitions 18 separated from each other by spacers 20 divide interior chamber 12 into a sequence of compartments 22 . each compartment 22 contains a dosage of the therapeutic , additive or nutrient substance to be released into the reticulorumen of the ruminant . spacers 20 are attached to or fit closely against the inner wall of body 10 . spacers 20 comprise rings made from paper or some other cellulosic material which dissolves in the reticulorumen environment . partitions 18 comprise solid discs of substantially the same diameter as interior chamber 12 . partitions 18 are held in place by spacers 20 . alternatively , spacers 20 may be omitted . partitions 18 will then be held in place by their close fit against the inner wall of body 10 and by the substance placed inside compartments 22 . partitions 18 are made from a material which is degraded upon exposure to the reticulorumen environment . for example , the partitions may be made from paper . the partitions retain the dosages in their compartment until they are exposed to the reticulorumen environment . the specific material selected determines the frequency of delivery of dosages . table i illustrates the relative degradability of partitions made of various materials in the reticulorumen of beef cattle . table i______________________________________ partition days / partition degraded______________________________________wax paper 5 . 4resin treated paper 1 . 540 lb . basis kraft paper 1 . 375 lb . basis kraft paper 1 . 0paraffin 0 . 4______________________________________ table i demonstrates that dosages of the therapeutic , additive or nutrient substance will be delivered daily into the reticulorumen of beef cattle if the partitions are made from a kraft paper of 75 lb . basis . retaining means are needed to retain the bolus within the ruminant . for example , weight means which impart to the bolus a specific gravity of at least about 1 . 5 , preferably about 1 . 5 - 7 . 0 , can serve as the retaining means . in the embodiment shown in fig1 a steel ball 24 serves as the weight means . the steel ball 24 is attached to the closed end 16 of body 10 . for example , the steel ball 24 may be enclosed within an epoxy shell as illustrated . the bolus should be small enough to pass readily into the reticulorumen of the particular ruminant . the bolus of fig1 should be no larger than 25 mm diameter and 100 mm long for administration to a calf weighing 180 kg . a bolus of this size would readily pass into the calf &# 39 ; s reticulorumen . the dimensions of each of the compartments 22 may also be varied to contain the desired dosage of the substance to be released . compartments that are 1 mm deep and 18 mm in diameter , as viewed in fig1 are adequate to deliver a daily dose of 300 mg rumensin ® to a calf weighing 180 kg . thus , the bolus of fig1 may be used to deliver approximately 90 daily doses of rumensin ® to growing calves . in use , the bolus is administered orally to the ruminant , for example , by means of a common balling gun apparatus . the bolus passes through the esophagus into the reticulorumen of the ruminant . due to its high specific gravity , it settles into the dorsal reticulorumen and is retained there . the body or shell 10 protects the partitions from bacterial attack except from the direction of open end 14 . the first partition is degraded or eroded by the bacteria , opening the first compartment and releasing a dose of the therapeutic , additive or nutrient substance contained therein . upon opening of the first compartment , the second partition becomes exposed to bacterial attack from the reticulorumen environment . the process repeats itself until all doses have been released at regular intervals . the material selected for the partitions determines the frequency of dosage release . when all the doses have been released , the bolus remains as a hollow shell inside the animal . the bolus can be allowed to remain there without causing any problems . if desired , each twentieth spacer can be made of a relatively digestible wood , such as oak wood , and may be left free from the epoxy coating . the shell will then break into smaller units and will leave the digestive tract after the bacteria degrade or erode the wooden spacers . fig3 illustrates a second embodiment of a bolus adapted to deliver therapeutic , additive or nutrient substances into the reticulorumen of a ruminant . the device shown in fig3 is similar to the one shown in fig1 and 2 except that a metal shell 30 , such as a stainless steel shell , serves as the weight means to impart the desired specific gravity to the bolus . inside the metal shell is an epoxy coating 32 , spacers 34 , and partitions 36 which divide interior chamber 38 into compartments 40 . in the embodiment shown , end 42 is open while end 44 is closed . bacterial attack can come only from the direction of open end 42 . alternatively , both ends can be open and bacterial attack can come from both ends . the device works in a manner similar to that of the device illustrated in fig1 and 2 . in addition to feed additives , such as rumensin ®, the device of the present invention may be used to dispense other substances , or combinations of substances , into the reticulorumen of a ruminant . these other substances include the following : rumen fermentation manipulators and ionophores , such as products sold under the trade names lasalocid , virginamycin , and ronnel , viable yeast cultures , sodium bicarbonate , mineral salts , zeolite , salinomyocin , and sarsoponin ; minerals , especially in regions of acute mineral deficiency , such as phosphorus on the gulf coast , copper in florida , magnesium in areas of grass tetany , and iodine in goiter areas ; pharmaceuticals for the treatment of specific diseases , such as antibiotics and sulfa drugs ; anthelmintics and other drugs , such as sedatives , antipyretics , hormones , and antibloating agents ; fecal markers and &# 34 ; rumen - by - pass &# 34 ; materials . devices were constructed similar to the one illustrated in fig1 and 2 . each bolus was fabricated with partitions made of 75 lb . basis uncoated kraft paper . each bolus was 100 mm long , 25 mm in diameter , and had 90 compartments . each compartment was loaded with a dosage of rumensin ® available from elanco products co . commercially available rumensin ® is prepared by extending a concentrated form of the active ingredient with rice hulls . commercially available rumensin ® is so bulky that recommended levels of the active ingredient cannot be provided in a bolus of the size fabricated . in this example , each compartment contained a bulky ( 60 g ./ lb .) powdered rumensin ® composition having 19 . 5 mg of active ingredient . twenty - four heifers averaging 625 lbs . were chosen for testing . the daily diet of each heifer comprised 6 lbs . of corn , 1 . 7 lbs . of a protein supplement , and orchard grass and white clover hay ad libitum . the heifers were kept on this diet for 86 days . a group of eight heifers , group a , received two devices at the start of the test . each bolus was dipped in mineral oil and administered orally to the heifers with a speculum . the heifers of group a were thus scheduled to receive from the devices 39 mg of rumensin ® per day . a second group of eight heifers , group b , received 300 mg of rumensin ® each in their daily supplement . the third group of eight heifers , group c , was the control group and the heifers received no rumensin ® in their diet . each animal &# 39 ; s pen was searched daily to see if a bolus had been regurgitated or passed . remnants of two devices were found in the pens of two animals on the 28th and the 38th days of the test respectively . inspection of the remnants showed that some of the epoxy had eroded and that the devices had lost their integrity . epoxies more resistant to erosion then the one employed are available commercially . the heifer that lost her devices on the 28th day was given two new ones on the 38th day . the averaged results of the tests are presented in table ii . table ii__________________________________________________________________________ group b ( no group c ( no group a ( bolus - group a heifers bolus - bolus - no no rumensin ® in that retained rumensin ® in rumensin ® in supplement ) their devices supplement ) supplement ) __________________________________________________________________________number of heifers 8 6 8 8initial weight ( lbs .) 630 598 623 621daily gain in weight 1 . 79 1 . 92 1 . 90 1 . 66for 86 days ( lbs ./ heifer / day ) daily gain in weight 1 . 42 1 . 57 1 . 29 0 . 97for final 33 daysof test ( lbs ./ heifer / day ) total hay consumption 822 837 593 789for 86 days ( lbs . ) feed efficiency 5 . 34 4 . 88 3 . 70 5 . 59for 86 days ( lbs . hay / lbs . gained ) __________________________________________________________________________ the daily gain in weight is measured more accurately than the feed efficiency and thus provides a better test for the utility of the device of the present invention . the data show that the heifers receiving rumensin ® from the devices had a higher daily weight gain than the heifers which did not receive rumensin ® ( group c ). this was true during the entire test period and during the last 33 days of the test . the performance during the last 33 days shows that the devices were still functional at that time . the data also show that the six heifers that retained their devices throughout had a daily weight gain which was similar to or better than the heifers which received rumensin ® in their supplement . again , this was true for the entire length of the trial and for the last 33 days of the trial . the improved performance of the six heifers that retained their devices over the entire group a heifers , including the two that lost their devices , also demonstrates the usefulness of the bolus . the increased daily weight gain experienced by heifers that received rumensin ® either by bolus or by supplement is within the range observed by other workers . feed efficiency is calculated as the number of pounds of hay consumed per pound of weight gain . thus , the lower the number , the more effective is the diet . the data for feed efficiency is not as significant as the data for daily weight gain . one reason is because calves have a tendency to throw hay out of their troughs . nonetheless , the data for feed efficiency show that heifers receiving rumensin ® through the bolus or from the supplement utilize their feed more effectively than heifers not receiving rumensin ®. it has previously been shown that rumensin ® does not influence feed utilization unless given to animals in a controlled manner daily or at least on alternate days . see muller , potter , and grueter , &# 34 ; alternate day vs . daily feeding of supplements containing monensin to pasture cattle ,&# 34 ; abstract from a . s . a . s . meeting at cornell univ ., july 27 - 30 , 1980 . the results of the tests described herein show that a device according to the present invention releases rumensin ® at the desired dosage rate . in further research , it has been found that under certain diet conditions , such as 100 % grass hay fed ad libitum , the bolus becomes clogged with hay particles after about 10 days . this slows the rate at which the partitions are degraded . placing a nylon cloth across the open end will prevent this from occurring by allowing microbes to pass through but not hay particles . while the invention has been described by reference to specific embodiments , this was for purposes of illustration only and should not be construed to limit the spirit and scope of the invention .

a device and a method for the prolonged administration of controlled amounts of a substance are disclosed . in one embodiment , the device is disclosed for the prolonged administration of dosages of a therapeutic , additive , or nutrient substance into the reticulorumen of a ruminant , comprising a body having an interior chamber , at least one end of said chamber open to the exterior of said body , said body made from a material which maintains its integrity in the reticulorumen environment , a plurality of degradable , such as cellulosic , partitions within said interior chamber connected to or fitting closely against the inner walls of said body , said partitions dividing said interior chamber into a series of compartments , each compartment containing a dosage of the therapeutic , additive or nutrient substance , said partitions arranged in an order to be degraded sequentially upon exposure to the reticulorumen environment thereby releasing sequentially the dosages of the substance into the reticulorumen , and retaining means provided by said body to retain the device within the reticulorumen of the ruminant .

as is shown in fig3 and 4 , the heterologous total stentless mitral valve prosthesis to be used in mitral and tricuspid position of the present invention is illustrated . this mitral valve prosthesis is comprised of a mitral sewing rim ( 1 ), a complete subvalvular apparatus ( 2 ) with chordal attachments ( 3 ) which are secured naturally between mitral leaflets ( 6 ) and heterologous portions or slices ( 8 ) of the papillary muscle ( chordal origin ) of the animal . as shown in fig4 these heterologous portions ( 8 ) of papillary muscle are fixed by suturing to a heterologous pericardial patch graft ( 5 ) or to a synthetic material to preserve its spatial geometry . the process of attaching the heterologous pericardial patches ( 5 ) to the chordal origins ( 8 ) is further described in fig5 - 10 . fig5 shows a harvested heterologous mitral heart valve ; the heart valve is harvested to preserve its anatomical integrity . the pericardial tissue ( 10 ) which is used to form pericardial patch ( 5 ) is also shown in fig5 . the pericardial tissues ( 10 ) are formed 20 % larger and in the same shape as their corresponding chordal origins ( 8 ). since the natural chordal origin is preserved , the spatial geometry between each chordae or chordal group is not violated and the natural angle of the chordal origin ( 8 ) is maintained . fig6 illustrates how the chordal origin or heterologous papillary muscle slice ( 8 ) is attached to the pericardial tissue ( 10 ) with interrupted and continuous sutures ( 15 ), such that the original papillary muscle angulation , as the chordae arise from the papillary slice ( 8 ), is preserved . the pericardial tissue ( 10 ) is made larger than the chordal origin ( 8 ) so the free pericardial edge can be used to anchor the mitral valve prosthesis to the patient &# 39 ; s papillary muscle . fig7 shows the sutures ( 15 ) in more detail to demonstrate that the sutures are not attached to the delicate chordal structure so that the structural integrity of the chordae are preserved . attaching the sutures directly to the chordal structure weakens the chordae and , therefore , lessens the durability of the mitral valve prosthesis . thus , by attaching the sutures to the pericardial tissue , rather than each chordae , the chordal structure is not weakened as it was by the prior art attachment methods . the pericardial tissue ( 10 ) is attached to the chordal origin ( 8 ) to preserve the original ( natural ) angulation of the chordal origin , which ensures that there will be uniform tension on the chordae when the mitral valve prosthesis is attached to the patient &# 39 ; s papillary muscle . by maintaining uniform chordal tension , undue stress on the chordal origin and on the individual chordae is avoided , which lessens the chance of papillary muscle tear and makes the mitral valve prosthesis more durable . since each mammal &# 39 ; s mitral valve has a different spatial geometry and natural chordal origin angulation , preserving the chordal origin is the best way to ensure that these structures are preserved , so that the valve prosthesis will obtain the advantages provided by uniform chordal tension . fig8 - 10 illustrate the attachment of an upper ( second ) pericardial tissue ( 11 ) to the chordal origin ( 8 ) and the pericardial tissue ( 10 ) to form the pericardial patch ( 5 ) of fig3 and 4 . the second pericardial tissue ( 11 ) is of the same size or smaller than the chordal origin ( 8 ) and is divided or sliced so as to fit around each chordae or chordal group , as shown in fig8 . the upper pericardial tissue ( 11 ) is attached , as shown in fig8 and 9 , on top of the heterologous papillary muscle slice ( 8 ) to both the chordal origin ( 8 ) and pericardial tissue ( 10 ) by continuous sutures ( 16 ). this attachment provides a well - secured chordal origin without any weakening of the chordal structure itself ; the sutures are not attached to the chordal structure . the second pericardial tissue ( 11 ), like the pericardial tissue ( 10 ), is attached to the chordal origin ( 8 ) so that the spatial geometry and natural angulation of the chordal origin is preserved . thus , the benefits provided by uniform chordal tension are maintained even after the second pericardial tissue ( 11 ) has been attached . fig1 illustrates the final affixation of the chordal origin to the pericardial patch ( 5 ). the chordae are surrounded by the upper pericardial tissue ( 11 ) so the sutures ( 17 ) do not injure the structure of the chordae and the chordal origin is attached to the pericardial tissues ( 10 and 11 ) with sutures ( 18 ) to fully respect the natural geometry of the chordal origin , thus the pericardial patch ( 5 ) is formed . the second ( upper ) pericardial tissue ( 11 ) acts as a collar to strengthen the chordal origin attachment and to avoid tearing of each chordae or chordal group from the thin heterologous papillary slice ( 8 ). the free edge of the pericardial tissue ( 10 ) is maintained throughout the procedure to allow the mitral valve prosthesis to be anchored to the patient &# 39 ; s papillary muscle . the free edge also provides a way to control alignment of the annulus ( described below ) and to address the issue of subvalvular height to attain consistent results . proper alignment and the correct subvalvuar height also helps to avoid undue stress and tension on the chordae and so the valve is more durable . to attach the pericardial patch ( 5 ), including the chordal origin ( 8 ), to the patient &# 39 ; s papillary muscle , it is preferred to use &# 34 ; u &# 34 ; type pledgetted sutures ( 18 ). the &# 34 ; u &# 34 ; type pledgetted sutures are squired at the origin of each papillary muscle ( 40 ) and are directed toward the papillary muscle head at a 45 degree angle towards the opposite side of the muscle , as shown in fig1 - 16 . the pledgetted sutures must never be placed close to the head of the papillary muscle because they will tear the muscle , as illustrated in fig1 b . the anterior sutures anchor the posterior edge of the pericardial patch ( 5 ) and the posterior sutures anchor the anterior edge of the pericardial patch ( 5 ) to the origin of the patient &# 39 ; s papillary muscle , as shown in fig1 - 16 . the suture spacing is widened as they extend from the origin of the papillary muscle to encompass the circumference of the pericardial patch ( 5 ) when the sutures are attached thereto , as shown in fig1 a . the pledgetted sutures are attached to the free edge of the lower pericardial tissue ( 10 ) of the pericardial patch ( 5 ) so as to respect the alignment of the chordal implant in relation to the left ventricular outflow tract , as shown in fig1 . the alignment of the mitral valve prosthesis is perfected in relation to the x markings on the right and left trigone of the implant valve ( described later ). the papillary muscle is analyzed to determine the optimal positioning of the pledgetted sutures at the posterior and anterior of the origin of the papillary muscle . it is preferred that 3 or 4 sutures be attached to each of the posterior and anterior portions of the patient &# 39 ; s papillary muscle to ensure the proper attachment and alignment of the chordal origin ( 8 ) and the entire mitral valve prosthesis . by following this procedure , repetitive good results are possible where they have never been known before . this heterologous device obtained from any mammal valve obtained from any mammal of size , anatomy and functions similar to the human heart , including , for example , swine , ovine , bovine and others . this heterologous mitral device is removed immediately after slaughtering of the animal by surgical technique . the entire mitral device is obtained and immersed into a balanced electrolyte , glutaraldehyde containing solution . thereafter , the excess of muscle tissue is removed by a dissection procedure . each procedure is performed so as to preserve the integrity of the mitral device . continual monitoring of the quality control of each element is done daily until the proper tanning and cross linking of the tissue is obtained by the balanced electrolyte glutaraldehyde containing solution . the posterior circumference of the patient &# 39 ; s mitral annulus may be normal , enlarged or distended or the diameter of the mitral annulus may be moderately enlarged depending on the etiology causing the valve stenosis , insufficiency or mixed lesion . thus , it becomes impossible to determine the correct stentless heterologous ( or homologous ) valve size , which is one of the reasons why previous techniques failed . if a valve with a smaller diameter is inserted into an enlarged mitral annulus , then the thin posterior annular circumference will rupture . even if the correct valve size is selected for implant , misalignment of the valve will produce mitral leaflet prolapse which creates valve insufficiency . prolapse can also be caused by incorrect valve sizing . therefore , it is important not only to select the correct size valve for implant but to strengthen the posterior annular circumference and provide a systematic way to align the valve upon implantation . fig1 - 13 illustrate the attachment of a pericardial tissue collar to the mitral valve sewing rim to strengthen the annulus , to provide for height adjustment of the valve in the patient and to provide a surface so that markings may be placed to determine correct sizing and alignment of the mitral valve . fig1 shows the heterologous mitral valve with the chordal origin attachment of the invention . the arrow ( 20 ) points to the naturally weak posterior circumference of the annulus . the weak posterior portion represents 2 / 3 of the annular circumference . once the valve is removed from the donator animal , the weak posterior portion of the annulus expands , making size determination impossible . thus , by placing a collar around the annulus it is returned to its correct size and the left and right trigone can be marked to allow selection of a properly sized valve . fig1 shows the attachment of a pre - formed pericardial tissue ( 21 ) over the heterologous mitral annulus . the pericardial tissue is preferably approximately 1 cm in width and of the same length as the circumference of the mitral annulus , thus the mitral annulus is reinforced with approximately 50 mm of biological material on either side once the pericardial tissue is attached thereto . the attachment of the pericardial tissue ( 21 ) serves many functions , including : maintaining the natural heterologous mitral valve diameter and circumference , reinforcing the naturally weak posterior circumference of the annulus , providing a surface to secure the mitral annulus to the patient &# 39 ; s mitral rim , permitting markings used to determine correct valve size and alignment ( illustrated in fig1 and described below ), and providing an extra approximately 20 or 30 mm of height adjustment for valve placement . the pericardial tissue collar ( 21 ) is attached to the mitral annulus by folding it around the annulus , thus enveloping the annulus , and using interrupted and / or continuous sutures ( 22 ) to attach the pericardial collar ( 21 ) to the mitral annulus , as shown in fig1 , with a needle ( 23 ). thus , the heterologous mitral annulus is placed between the pericardium so that the above - mentioned advantages may be achieved . the new sewing ring is completely biological , with the exception of the sutures which are mono or polyfilament sutures . fig1 illustrates the markings used to determine correct valve size and provide a reference for the structural alignment required during valve implant . the x markers are placed at each of the heterologous mitral valve trigone ( 25 ). the distance between the x markers must match the patient &# 39 ; s intertrigonal width for the valve size to be correct . the space between the left and right trigone is used because it does not vary significantly ( little or no change ) in either the patient or the heterologous mitral valve . the type of disease affecting the patient &# 39 ; s mitral valve does not affect the size of this area and the anterior annular portion of the heterologous mitral valve is strong and fibrous and so does not vary in size as does the weak posterior portion . the x markings ( 25 ) are made with color sutures on the pericardial collar ( 21 ) and provide a fixed distance to determine valve size . the x markings are also used in combination with the edge of pericardial tissue ( 10 ) to help attain proper alignment of the valve upon implantation . proper alignment of the valve prosthesis allows a uniform tension to be maintained when the chordae through pericardial patches ( 5 ) are attached to the patient &# 39 ; s papillary muscle and thus the durability of the mitral valve prosthesis is increased . the midpoint of the posterior annular circumference is marked &# 34 ; i &# 34 ; ( 26 ) to help determine the correct valve size and maintain alignment of the mitral valve prosthesis during implant . the &# 34 ; i &# 34 ; marking ( 26 ) further enables the valve to be implanted so that the stress force is more evenly distributed during the attachment of the valve to avoid injuring the weak posterior of the annulus . as an alternative to the pericardial tissue collar shown in fig1 - 13 , a pericardial tissue collar ( 30 ) containing naturally secured vertical pericardial strips ( 31 ) can be used , as shown in fig1 - 18 . the pericardial collar ( 30 ) of fig1 - 18 provides all of the above mentioned advantages and , furthermore , it decreases the stress on the chordae ( 3 ) by 50 % through the attachment of the pericardial strips ( 31 ) shown in fig1 - 18 . the pericardial strips ( 31 ) are part of the same tissue used to create the pericardial collar ( 30 ). these strips ( 31 ) are attached to pericardial patch ( 5 ) with continuous and interrupted sutures ( 33 ), as shown in fig1 , to divide the stress load over more points . thus , instead of the chordae ( 3 ) absorbing all the stress of attachment , these pericardial strips ( 31 ) absorb 50 % of the stress , which leads to a more durable valve prosthesis . the pericardial strips ( 31 ) are attached to the pericardial patch ( 5 ) and not the chordae ( 3 ), thus the chordal structure is not weakened . these strips are attached to the pericardial patch ( 5 ) and the chordal origin ( 8 ) so that the natural angulation and spatial geometry of the chordal origin ( 8 ) and chordae are maintained . the strips are the same length as the chordae of the valve prosthesis to maintain a uniform chordal tension and thus properly distribute the stress load between the strips and the chordae to increase the valve prosthesis &# 39 ; durability . in the device of the present invention each individual chordae and chordal group ( 3 ) is preserved and naturally secured between mitral leaflets ( 6 ) and heterologous portions ( 8 ) of papillary muscle . these portions ( 8 ) of papillary muscle are affixed between two heterologous pericardial patches or synthetic materials using mono or polyfilament sutures , as shown in fig5 - 10 . as shown in fig4 the pericardial patches ( 5 ) are then sutured to the papillary muscle ( 4 ) of the patient . this procedure properly anchors the chordal groups to the papillary muscle ( 4 ), while respecting the spatial geometry and natural angulation of each chordae or chordal group ( 3 ). further , a biological pericardial ( 21 or 30 ) collar can be sewn to the mitral sewing rim and this collar may further include various vertical heterologous strips ( 31 ) which are attached to the pericardial patch ( 5 ) to help distribute the stress of attachment . this total biological device consists of a sewing collar ( 21 or 30 ), leaflets ( 6 ) and the subvalvular apparatus ( 2 ), having chordal attachments ( 3 ) which are naturally secured to their chordal origin ( 8 ), which is affixed to the pericardium ( 5 ) or to synthetic material and used as a heart valve substitute for the mitral and tricuspid valve of humans . this device presents a specific design that allows the anchoring of it to the papillary muscle and mitral annulus of the patient , thus providing for better hemodynamic performance and avoiding disfunction and dilatation of the left ventricle . as a result thereof , this device benefits the recipient &# 39 ; s quality of life as well as the durability of the device during use . this heterologous total stentless mitral heart valve prosthesis may provide better durability , since it has all the known elements necessary for the harmonic function of the mitral valve and left ventricle which are not present in previous heart valve prostheses . while in the foregoing specification a detailed description of an embodiment of the invention has been described for the purpose of illustration , many variations in the details herein given may be made by those skilled in the art without departing from the spirit and scope of the invention .

the present invention relates to a heterologous total stentless mitral valve prosthesis for the use in mitral and tricuspid position which provides for better hemodynamic performance . this mitral valve prosthesis addresses the specific needs of flow and contractility , thereby prolonging durability . the heterologous total stentless mitral heart valve prosthesis of this present invention comprises one sewing ring , leaflets , and subvalvular apparatus having new chordal insertions which are fixed to or in between two heterologous pericardial patches . this heart valve prosthesis is obtained from heterologous mammals , such as swine , ovine , bovine , and moreover , the pericardial patches used may also be of synthetic material . the heart valve prosthesis presents specific design benefits from its anatomical and functional characteristics permitting chordal insertion to the papillary muscles and attachment of its sewing rim to the mitral annulus of the recipient , thereby providing for better hemodynamic performance and avoiding disfunction and dilatation of the left ventricle .

the present invention is directed to methods and devices for performing percutaneoous surgery and in particular spinal surgery . the surgery is performed through a portal or passageway provided by a retractor . the retractor is expandable in situ to thereby increase the size of the surgical area as well as the access thereto . it is particularly constructed to minimize trauma to tissue surrounding the retractor and the surgical area . the retractor can be used with any surgical approach to the spine such as ; lateral , postero - lateral and / or antero - lateral , anterior , posterior , posterior mid - line , and in other regions of the body not associated with the spine . fig1 is a top view of retractor 1 . as shown retractor 1 is comprised of four discrete segments . as shown , the segments consist of a pair of opposing semi cylindrical members 4 a and 4 b as well as a pair of opposing semi elliptical members 6 a and 6 b . a ratcheting mechanism 8 a and 8 c is located at each of the junctions between opposing semi cylindrical member 4 b and opposing semi elliptical members 6 a and 6 b . likewise a ratcheting mechanism 8 b and 8 d is located at each of the junctions between semi cylindrical segment 4 a and opposing semi elliptical members 6 a and 6 b . the four segments once assembled and surrounded by a silicone sleeve 10 and form a single working unit that is generally elongated and oval shaped in cross section . the sleeve 10 conforms to the shape of the exterior surfaces of the segments and extends the entire length of the segments from the proximal end , the top portion , to the distal end , bottom portion and exerts a radially directed inward force against segments 4 a , 4 b , 6 a and 6 b . the assembly thereby forms an elongated ovoid shaped retractor wherein the parallel distraction will occur along the length of the retractor based upon the engagement and disengagement of the teeth placed along the longitudinal axis of the retractor . while shown and described as having four segments the retractor could be formed as two segments each including a semi cylindrical segment and a semi elliptical segment . the segments 4 a , 4 b , 6 a and 6 b are formed from plastic or any other suitable radio lucent material . segments 4 a and 4 b each respectively have screw holes 12 a and 12 b designed to receive a bone screw for distal fixation of the retractor to a vertebral body . the segments also contain insulated electrical conductors 16 included in the walls of the segments . the conductors 16 terminate at the proximal and distal surfaces of the segments with exposed electrical contacts to provide an electrical pathway for nerve monitoring also included within the segments are internal tracks for mounting fiber optical lights 18 to provide illumination of the surgical space located at the distal end of the retractor . each of the segments 4 a , 4 b , 6 a , and 6 b may contain radio opaque markers 24 to enable visualization throughout the procedure . fig2 shows a partially exploded top view if the four segments prior to engagement via the ratcheting mechanisms . fig3 is an enlarged view of the encircled area of fig2 as can be seen in fig3 semi - circular segments have teeth or grooves 20 located on the interior surface adjacent both edges of the semi circular segments 6 a and 6 b . these teeth or grooves 20 are located at four separate points along the length of the segments between the proximal and distal end portions as shown in fig5 . alternatively teeth or grooves 20 can run the entire length of the segments from the distal end portion to the proximal end portion as shown in fig6 . as seen from the front view , the lower distal end surface of segments 6 a and 6 b form a concave edge 26 . fig7 shows a side view of the external surface of the semi cylindrical segments 6 a and 6 b . fig8 is a front view of the exterior surface of one of the semi elliptical members 4 a and 4 b . as seen from the front view the lower distal end surface of segments 4 a and 4 b form a convex edge 28 . concave edges 26 and convex edges 28 form a tip at the distal end of the retractor 1 that is anatomical in shape and particularly configured to significantly improve the ability to mobilize , dissect , split and retract the terminal tissues of the psoas muscle at the point on the spine where the entry is to be made . fig9 is an exterior surface side view of one of the semi elliptical members 4 a and 4 b . one or both of the segments 4 a and 6 b contain an insulated electrical conductor 16 included in the walls of the segments . a tool 30 , shown in fig1 a and 11b is used to facilitate a ninety degree rotation of the retractor as will be explained below . the tool 30 includes a pair of diametrically opposed handles 32 and 34 that are each connected to an annular member 36 . the inner surface 38 of the annular member 36 is configured to operatively cooperate with the external surface of sleeve 10 surrounding the retractor 1 adjacent the top portion thereof . semi elliptical segments 4 a and 4 b have complimentary teeth or grooves 22 . teeth or grooves 22 are located on the exterior of semi elliptical segments 4 a and 4 b adjacent each of the edges of semi elliptical segments 4 a and 4 b . teeth or grooves 22 extend the entire length of the segments form the proximal to distal end portion as shown in fig9 . fig4 shows one of the ratcheting mechanisms 8 a - 8 d and the inter engagement of teeth 20 and 22 are well as sleeve 10 which exerts a radially directed inward force on each of the retractor segments . the present system is a patient based retractor that does not require fixation to an articulating arm . the patient based retractor includes a pad 40 that eliminates the necessity for a rigid fixation to a point outside of the field of operation or to an independent immobile point such as a bed rail . the pad 40 is applied to the sterilized area on the patient &# 39 ; s body 41 . the physical properties of the material including its size , thickness and composition cause significant friction , or adhesion , between the pad and the sterile site on the patient &# 39 ; s skin . by way of example , fig1 a and 10b illustrate the pad 40 in a deployed position . this can be reinforced with the application of tape and or ioban ® if necessary . by way of example the pad can be formed from a polyurethane material . while the pad 40 and retractor 1 have been shown for use during minimally invasive spinal surgery it should be understood that the anchoring pad 40 could be used in combination with retractor 1 or a retractor of any configuration and for other types of surgery as well , such as laparoscopic gal bladder surgery or appendectomy . the tube retractor technique starts with identifying the correct entry point , and correct trajectory from the skin to the pathology to be addressed . a top view of the incision 42 is shown in fig1 b . the initial soft tissue canulated dilator 50 of circular cross section , as shown in fig1 a , is inserted through the incision and forcefully advanced to the objective site . a series of larger diameter dilators as shown in fig1 b , 51 and 52 , are inserted over the initial dilator sequentially increasing in diameter until the final operative diameter is inserted . the final operative dilator 54 is oblong in cross section as is shown in fig1 c through 12f . fig1 c is a front perspective view of the ovoid shaped final dilator . fig1 d is a side view of the bottom or distal end surface 56 of the final dilator that is convex in shape on both side wall portions . fig1 e is a perspective bottom view of the bottom or distal end surfaces of the final dilator that shows the front and back wall surfaces having bottom edges 58 that are concave in shape . the two convex surfaces at the lower edge of the side wall portions and the concave edges on the front and back walls form a distal or end surface that is anatomical in shape and particularly configured to significantly improved the ability to mobilize , dissect , split and retract the terminal tissues of the psoas muscle at the point on the spine where the entry is to be made . once in its proper position the oblong operative dilator 54 is rotated ninety degrees and then counter rotated back to its original position using the tool 30 . fig1 is a top view of the retractor in position on the patient . fig1 a is a top view of the retractor 1 in position within the patient &# 39 ; s body and fig1 b is a side view of the retractor within the patient &# 39 ; s body 41 . as shown in fig1 a and 14b the spinal disc 60 is located between vertebral bodies 62 and 64 . as shown , each vertebral body includes a spinous process bone 66 and a pair of pedicle bones 68 . either segment 6 a or 6 b can be seen in this view . once in this position , the retractor 1 is then rotated ninety degrees using tool 30 to the position shown in fig1 . fig1 is a top view of the retractor 1 in position on the patient after being rotated . the shape of the distal end of the retractor segments provides a significant improvement in the ability to mobilize , dissect , split and retract the terminal tissues of the psoas muscle at the point on the spine where entry is to be made . current designs are parallel to the spine and do not comply with the natural shape of the spine . thereby allowing the terminal psoas muscle fibers to creep under the retractor and completely undermine the process and in many cased reduces the overall success and intention of the minimally invasive technique . the ninety degree rotation of the retractor 1 enables the distal portions of the psoas muscle to be mobilized and retracted via the retractor . this action reduces muscle creep thereby reducing the necessity for the surgeon to cut , cauterize and remove muscle fibers to access to the pathology . once the retractor 1 is rotated into final position the final and initial dilators are withdrawn from the patient . at this point , a series of retractor expansion dilators , shown in fig1 c , are available for expansion of the retractor 1 . these expansion dilators are canulated and assist in centering the retractor with the initial k - wire that is already in place . the expansion dilators are 14 , 16 , 18 and 20 mm in diameter . as progressively larger expansion dilators are inserted within the retractor 1 , segments 4 a , 4 b , 6 a and 6 b move relative to one another by virtue of ratcheting mechanisms 8 a , 8 b , 8 c and 8 d . by way of example , it is contemplated that the distance between segments 4 a and 4 b at their mid points can be increased from 14 mm to 18 mm and the distance between segments 6 a and 6 b at their mid points can be increased from 18 mm to 22 mm , as shown diagrammatically in fig1 a and 17b . at this point the expansion dilators are removed and the operation can proceed . the retractor is anchored at the near portion by frictional engagement with pad 40 . in addition the retractor 1 may be anchored at the distal end portion using bone screws that are inserted through holes 12 a and 12 b of the retractor and threaded into the vertebral body . the pad 40 eliminates the need for a rigid fixation to a point outside the surgical field or to an independent fixed point such as a bed rail . the elimination of these metallic supports that are typically associated with minimally invasive tube or split blade retractors increases the visualization of the approach through the retractor and visualization of the surgical site while using operative fluoroscopy . the retractor system of the present invention was developed to provide minimally invasive access to a patient &# 39 ; s pathology . the ability to dilate muscle tissue , as opposed to the process where the muscle tissue to stripped or detached from the skeletal structure will usually reduce the morbidity associated with the standard invasive technique . the procedure to utilizing the retractor system of the present invention starts with the identification of the correct entry point , the proper trajectory from the skin to the pathology to be addressed as well as the point of incision . after the initial incision has been made through the patient based anchoring pad and into the skin the initial soft tissue dilator is inserted through the incision and forcefully advanced to the objective site . the initial dilator is 6 mm in diameter and round in cross section . thereafter , a series of progressively larger dilators are inserted over the initial dilator ; increasing in diameter until the final operative dilator is inserted . the final operative dilator is oblong in cross section . once the final operative oblong dilator is in place it is then rotated ninety degrees by tool 30 and then counter rotated ninety degrees back to its initial position . the retractor 1 is then placed over the final operative dilator and forcefully advanced to the objective site . once in position the retractor is then rotated ninety degrees by using a tool 30 . thereafter the initial and final dilators are removed . following removal of the dilators used for initial delivery , a series of expansion dilators , are inserted into the center of the retractor 1 to expand the open area or portal within the retractor . these expansion dilators are circular in cross section and range in diameter from 14 mm to 20 mm . as the expansion dilators are inserted the ratcheting mechanisms 8 a , 8 b , 8 c , and 8 d allow relative movement between the adjacent retractor segments by virtue of the disengagement and reengagement of the teeth 20 and 22 . the expansion dilator creates a force directed radially outwards thereby causing a shift in the alignment of teeth 20 and 22 . simultaneously resilient sleeve 10 exerts a radially inward directed force maintaining the teeth 20 and 22 in their newly established position . all patents and publications mentioned in this specification are indicative of the levels of those skilled in the art to which the invention pertains . all patents and publications are herein incorporated by reference to the same extent as if each individual publication was specifically and individually indicated to be incorporated by reference . it is to be understood that while a certain form of the invention is illustrated , it is not to be limited to the specific form or arrangement herein described and shown . it will be apparent to those skilled in the art that various changes may be made without departing from the scope of the invention and the invention is not to be considered limited to what is shown and described in the specification and any drawings / figures included herein . one skilled in the art will readily appreciate that the present invention is well adapted to carry out the objectives and obtain the ends and advantages mentioned , as well as those inherent therein . the embodiments , methods , procedures and techniques described herein are presently representative of the preferred embodiments , are intended to be exemplary and are not intended as limitations on the scope . changes therein and other uses will occur to those skilled in the art which are encompassed within the spirit of the invention and are defined by the scope of the appended claims . although the invention has been described in connection with specific preferred embodiments , it should be understood that the invention as claimed should not be unduly limited to such specific embodiments . indeed , various modifications of the described modes for carrying out the invention which are obvious to those skilled in the art are intended to be within the scope of the following claims .

a retractor having an elongate body that provides access to a surgical location within a patient . the elongate body is generally ovoid in cross section and includes a plurality of segments that are connected to one another through a plurality of ratcheting mechanisms . the ratcheting mechanisms permit relative movement of the segments with respect to one another when expander dilators are inserted within the retractor . the segments are surrounded and retained by a resilient elastomeric sleeve . the distal end surfaces of the segments include edges that are configured to mobilize , dissect , split and retract the terminal tissues in the surgical area . the retractor is used in conjunction with a resilient elastomeric pad that is affixed to the patient and firmly engages the outer surface of the elongate body to thereby anchor the retractor to the patient .

several embodiments of the present invention will now be described in detail with reference to the accompanying drawings . fig1 shows a block diagram of an x - ray ct apparatus , which is an embodiment of the present invention . the configuration of the apparatus represents an embodiment of the apparatus in accordance with the present invention . the operation of the apparatus represents an embodiment of the method in accordance with the present invention . as shown in fig1 the apparatus comprises a scan gantry 2 , an imaging table 4 and an operating console 6 . the scan gantry 2 has an x - ray tube 20 . x - rays ( not shown ) emitted from the x - ray tube 20 are formed into a fan - shaped x - ray beam , i . e ., a fan beam , by a collimator 22 , and projected toward an x - ray detector 24 . the x - ray detector 24 has a plurality of detector elements arranged in line as an array in the extent direction of the fan - shaped x - ray beam . the configuration of the x - ray detector 24 will be described in detail later . the x - ray tube 20 , collimator 22 and x - ray detector 24 together constitute an x - ray emitting / detecting apparatus , which will be described in detail later . the x - ray detector 24 is connected with a data collecting section 26 . the data collecting section 26 collects signals detected by the individual detector elements in the x - ray detector 24 as digital data . the emission of the x - rays from the x - ray tube 20 is controlled by an x - ray controller 28 . the interconnection between the x - ray tube 20 and x - ray controller 28 is omitted in the drawing . the collimator 22 is controlled by a collimator controller 30 . the interconnection between the collimator 22 and collimator controller 30 is omitted in the drawing . the above - described components from the x - ray tube 20 through the collimator controller 30 are mounted on a rotating section 34 of the scan gantry 2 . the rotation of the rotating section 34 is controlled by a rotation controller 36 . the interconnection between the rotating section 34 and rotation controller 36 is omitted in the drawing . the imaging table 4 is configured to carry a subject to be imaged ( not shown ) into and out of an x - ray irradiation space in the scan gantry 2 . the relationship between the subject and x - ray irradiation space will be described in detail later . the operating console 6 has a data processing apparatus 60 . the data processing apparatus 60 is comprised of , for example , a computer . the data processing apparatus 60 is connected with a control interface 62 . the control interface 62 is connected with the scan gantry 2 and the imaging table 4 . the data processing apparatus 60 controls the scan gantry 2 and imaging table 4 via the control interface 62 . the data collecting section 26 , x - ray controller 28 , collimator controller 30 and rotation controller 36 in the scan gantry 2 are controlled via the control interface 62 . the individual connections between these sections and the control interface 62 are omitted in the drawing . the data processing apparatus 60 is also connected with a data collection buffer 64 . the data collection buffer 64 is connected with the data collecting section 26 in the scan gantry 2 . data collected at the data collecting section 26 are input to the data processing apparatus 60 via the data collection buffer 64 . the data processing apparatus 60 performs image reconstruction using transmitted x - ray data for a plurality of views collected via the data collection buffer 64 . the image reconstruction is performed using a filtered backprojection technique , for example . the data processing apparatus 60 is also connected with a storage device 66 . the storage device 66 stores several kinds of data , programs , and so forth . several kinds of data processing relating to imaging are achieved by the data processing apparatus 60 executing the programs stored in the storage device 66 . the data processing apparatus 60 is further connected with a display device 68 and an operating device 70 . the display device 68 displays the reconstructed image and other information output from the data processing apparatus 60 . the operating device 70 is operated by a user , and supplies several kinds of instructions and information to the data processing apparatus 60 . the user interactively operates the present apparatus using the display device 68 and operating device 70 . [ 0045 ] fig2 schematically shows one configuration of the x - ray detector 24 . as shown , this x - ray detector 24 is a multi - channel x - ray detector having a multiplicity of detector elements 24 ( i ) arranged in a one - dimensional array . reference symbol ‘ i ’ designates a channel index and ‘ i ’= 1 - 1 , 000 , for example . the detector elements 24 ( i ) together form an x - ray impingement surface curved as a cylindrical concavity . the x - ray detector 24 may instead be one having a plurality of detector elements 24 ( ik ) arranged in a two - dimensional array , as shown in fig3 . the detector elements 24 ( ik ) together form an x - ray impingement surface curved as a cylindrical concavity . reference symbol ‘ k ’ designates a row index and ‘ k ’= 1 , 2 , 3 , 4 , for example . the detector elements 24 ( ik ) that have the same row index ‘ k ’ together constitute a detector element row . the x - ray detector 24 is not limited to having four detector element rows , and may have a plurality of rows that is more or less than four rows . each detector element 24 ( ik ) is formed of a combination of a scintillator and a photodiode , for example . it should be noted that the detector element 24 ( ik ) is not limited thereto but may be a semiconductor detector element using cadmium telluride ( cdte ) or the like , or an ionization chamber detector element using xenon ( xe ) gas , for example . [ 0048 ] fig4 shows an interrelationship among the x - ray tube 20 , collimator 22 and x - ray detector 24 in the x - ray emitting / detecting apparatus . fig4 ( a ) is a view from the front of the scan gantry 2 and ( b ) is a view from the side thereof . as shown , the x - rays emitted from the x - ray tube 20 are formed into a fan - shaped x - ray beam 400 by the collimator 22 , and projected toward the x - ray detector 24 . [ 0049 ] fig4 ( a ) illustrates the extent of the fan - shaped x - ray beam 400 . the extent direction of the x - ray beam 400 coincides with the direction of the linear arrangement of the channels &# 39 ; in the x - ray detector 24 . fig4 ( b ) illustrates the thickness of the x - ray beam 400 . the thickness direction of the x - ray beam 400 coincides with the direction of the side - by - side arrangement of the detector element rows in the x - ray detector 24 . a subject 8 placed on the imaging table 4 is carried into the x - ray irradiation space with the subject &# 39 ; s body axis intersecting the fan surface of such an x - ray beam 400 , as exemplarily shown in fig5 . the scan gantry 2 has a cylindrical structure containing therein the x - ray emitting / detecting apparatus . the x - ray irradiation space is formed in the internal space of the cylindrical structure of the scan gantry 2 . an image of the subject 8 sliced by the x - ray beam 400 is projected onto the x - ray detector 24 . the x - rays passing through the subject 8 are detected by the x - ray detector 24 . the thickness ‘ th ’ of the x - ray beam 400 impinging upon the subject 8 is regulated by the degree of opening of an aperture of the collimator 22 . the x - ray emitting / detecting apparatus comprised of the x - ray tube 20 , collimator 22 and x - ray detector 24 continuously rotates ( or scans ) around the body axis of the subject 8 while maintaining their interrelationship . when the imaging table 4 is continuously moved in the body axis direction of the subject 8 as indicated by an arrow 42 simultaneously with the rotation of the x - ray emitting / detecting apparatus , the x - ray emitting / detecting apparatus will rotate relative to the subject 8 along a helical trajectory surrounding the subject 8 , thus conducting a scan generally referred to as a helical scan . it will be easily recognized that when a scan is conducted with the imaging table 4 immobilized , a scan at a fixed slice position , i . e ., an axial scan , is conducted . projection data for a plurality of ( for example , ca . 1 , 000 ) views are collected per scan rotation . the collection of the projection data is conducted by a system including the x - ray detector 24 , data collecting section 26 and data collection buffer 64 . when the number of detector element rows in the x - ray detector 24 is four , data for four slices are simultaneously collected , as shown in fig6 . the data processing section 60 uses the projection data for the four slices to perform image reconstruction . representing the distance between centers of adjacent slices as ‘ s ’, and the movement distance of the x - ray emitting / detecting apparatus in the body axis direction per rotation of a helical scan as ‘ l ’, l / s is generally referred to as the pitch of the helical scan . prior to such a scan , dose adjustment for the particular subject 8 is conducted . the dose adjustment is achieved by modulating the tube current - time product , i . e ., the so - called milliampere - seconds ( mas ), for the x - ray tube . the tube current - time product will be sometimes referred to simply as the tube current hereinbelow . tube current adjustment for the particular subject 8 is sometimes referred to as auto - milliampere ( auto ma ). for the tube current adjustment , a projection of the subject 8 is measured . the measurement of the projection is achieved by fluoro - imaging the subject 8 by the x - ray beam 400 in , for example , a 0 ° ( sagittal ) direction and a 90 ° ( lateral ) direction , and obtaining respective projections , as conceptually shown in fig7 . such fluoro imaging will be sometimes referred to as scout imaging hereinbelow . for these projections , respective projection areas are calculated by the equations below . the calculation is conducted by the data processing section 60 . the same applies to the following description . projection_area = ∑ i = 1 i = max_ch   proj 0   deg   i ,  and ( 1 ) projection_area = ∑ i = 1 i = max_ch   proj 90   deg   i , ( 2 ) proj 0degi : projection data for each channel in the sagittal direction , and proj 90degi : projection data for each channel in the lateral direction . the projection areas calculated using equations ( 1 ) and ( 2 ) will have the same value . for the sagittal and lateral projections , respective center values are calculated using the following equations : proj_  0   deg = ∑ i = cent - 49 i = cent + 50   proj 0   deg   i ,  and ( 3 ) proj_  90   deg = ∑ i = cent - 49 i = cent + 50  proj 90   deg   i , ( 4 ) cent + 50 : a number obtained by adding 50 to the center channel index , and cent − 49 : a number obtained by subtracting 49 from the center channel index . proj — 0deg will be sometimes referred to as a sagittal center value and proj — 90deg as a lateral center value hereinbelow . the center values are used to calculate an oval ratio when the cross section of the subject 8 is assumed to be elliptical . the oval ratio is given by the following equation : oval_ratio = ∑ i = cent - 49 i = cent + 50  proj 90   deg   i ∑ i = cent - 49 i = cent + 50   proj 0   deg   i . ( 5 ) it should be noted that the numerator and denominator of the oval ratio are set so that the oval ratio has a value no less than one . therefore , if the sagittal center value is greater than the lateral center value as in the head , the sagittal center value is set at the numerator and the lateral center value is set at the denominator , contrary to the equation above . the one of the sagittal and lateral center values that has a larger value corresponds to the major axis of the ellipse , and the other that has a smaller value corresponds to the minor axis . it also possible to obtain only one projection fluoro - imaged in either the sagittal or the lateral direction . in this case , the projection area is obtained from either equation ( 1 ) or ( 2 ) depending upon the direction of the fluoro - imaging , and the center value of the projection is similarly obtained from either equation ( 3 ) or ( 4 ) depending upon the direction of the fluoro - imaging . the relationship among the projection area , sagittal center value and lateral center value is given by the following equation : projection_area =( proj — 0 deg × proj — 90 deg )× s + i , ( 6 ) therefore , if any two of the projection area , sagittal center value and lateral center value are known , the remaining one value can be arithmetically determined . when the projection area and one of the center values are known from fluoro - imaging in one of the directions , the other center value is determined by the following equation : proj_orthogonal = projection_area - 1 proj_measure × s , ( 7 ) therefore , when proj_measure is the sagittal center value , the oval ratio is given by : and when proj_measure is the lateral center value , it is given by : it will be easily recognized that also in this case , the numerator and denominator are set so that the oval ratio is no less than one . the quality of a reconstructed image is represented by an image sd ( image standard deviation ). the image sd when the subject has a circular cross section is a function of the projection area under a certain reference dose , and is given by the following equation : α , β , γ : constants that depend upon the tube voltage ( kv ) etc . when the subject has an elliptical cross section , the image sd varies with the oval ratio . assuming that the projection area is constant , the relationship between the oval ratio and the rate of change of the image sd is given by the following equation : the relationship of equation ( 11 ) is shown by the graph in fig8 . as shown , when the oval_ratio is one , the sd ratio is one . that is , the image sd does not vary when the cross section is circular . from such a relationship , when the subject has an elliptical cross section , a modified image sd is determined for the shape of the cross section by the following equation : the modified image sd is a predicted value for the image sd of a reconstructed image when the subject 8 is imaged by a reference dose . since a target value of the image sd for the reconstructed image is determined beforehand , the dose must be set so that an image satisfying the target value is obtained . the relationship among the image sd predicted value and the reference dose , and the image sd target value and the required dose is given by the following equation : image_sd target image_sd predited = mas reference × thickness_factor mas scan , ( 13 ) thickness_factor = 10 . 0 thickness  ( mm ) . ( 14 ) the ‘ thickness ’ is the thickness of the x - ray beam 400 at the iso - center of the subject 8 . mas scan = mas reference × thickness_factor ( image_sd target image_sd predicted ) 2 . ( 15 ) thus , the tube current corresponding to the required dose is obtained as follows : ma scan = mas scan scan_time   ( sec ) , ( 16 ) where ‘ scan_time ’ is the scan time of the present apparatus , i . e ., the time period during one rotation of the x - ray emitting / detecting apparatus . [ 0103 ] fig9 shows a flow chart of the operation from the scout imaging to the tube current calculation as described above . as shown , scout imaging is conducted at step 502 . by the scout imaging , the subject 8 is fluoro - imaged in one or both of sagittal and lateral directions over a certain range in the body axis direction , and respective projections at positions on the body axis are acquired . next , at step 504 , localization is conducted . the localization is for specifying scan start and end points on the body axis in a fluoroscopic image obtained by the scout imaging . this determines the length of the imaged range , and for a helical scan , determines a scan position for every pitch . the localization is performed by the user via the operating device 70 . next , at step 506 , an image sd target value is input . the input is also performed by the user via the operating device 70 . if a standard value pre - stored in the present apparatus is used for the image sd target value , input by default is possible . next , at step 508 , an image sd is calculated . in calculating the image sd , a projection area is first obtained . when the scout imaging is conducted in the sagittal and lateral directions , respective projection areas are obtained from equations ( 1 ) and ( 2 ); and when the scout imaging is conducted in one of these directions , a projection area is obtained by equation ( 1 ) or ( 2 ) depending upon the direction . the image sd is then calculated from equation ( 10 ) using the projection area ( s ). the image sd is calculated for every pitch of the helical scan . calculations for values described below are also conducted in the same way . next , at step 510 , a modified image sd is calculated . prior to calculating the modified image sd , sagittal and lateral center values are obtained from equations ( 3 ) and ( 4 ), respectively , and an oval ratio is obtained from equation ( 5 ). alternatively , a sagittal or lateral center value is obtained from equation ( 3 ) or ( 4 ), a lateral or sagittal center value is obtained from equation ( 7 ), and an oval ratio is obtained from equation ( 8 ) or ( 9 ). an sd ratio is obtained from equation ( 11 ) using the oval ratio , and the sd ratio is used to calculate the modified image sd from equation ( 12 ). next , at step 512 , dose is calculated . the dose calculation is performed according to equation ( 15 ). in this equation , the image sd target value input at step 506 is used as image_sd target , and the two modified image sd &# 39 ; s obtained as described above are used as image_sd predicted . thus , two doses are calculated . next , at step 514 , a tube current is calculated . the tube current calculation is performed according to equation ( 16 ). the solid line in fig1 shows an exemplary tube current thus calculated . as shown , the tube current is obtained for every position on the body axis . the tube current indicated by the broken line will be explained later . next , at step 516 , the calculated values for the tube current are stored in the memory . thus , the tube current is stored for every pitch of the helical scan . a general operation of the present apparatus will now be described . fig1 is a flow chart of the general operation of the present apparatus . as shown , at step 602 , an upper limit of the exposure dose is specified . the specification is performed by the user via the display device 68 and operating device 70 . a portion comprised of the display device 68 and operating device 70 is an embodiment of the setting means of the present invention . the exposure dose is set using a dlp ( dose length product ). the unit for the dlp is mgy · cm ( milligray · centimeter ). the upper limit for the dlp is specified as , for example , 300 mgy · cm . the upper limit for the dlp will be sometimes referred to as dlpu hereinbelow . next , at step 604 , an imaging protocol is specified . when auto - milliampere is employed , the protocol specification is achieved by the operation as shown in fig9 . by auto - milliampere , a tube current for the particular subject 8 is set . when auto - milliampere is not employed , the imaging protocol desired by the user is specified , and the tube current is also set at a desired value . next , at step 606 , a predicted value for the exposure dose is calculated . the calculation of the exposure dose predicted value is achieved based on the tube current set value . specifically , based on the tube current set value , a ctdivol ( ct dose index volume ) is first calculated . the calculation of the ctdivol based on the tube current is executed using a predefined algorithm . alternatively , the ctdivol is found from a pre - measured relationship between the ctdivol and tube current using a phantom , for example . the unit for the ctdivol is mgy . the exposure dose predicted value is obtained by multiplying the ctdivol by the imaged length in the body axis direction . the exposure dose predicted value will be denoted by dlpc hereinbelow . it should be noted that the upper limit for the exposure dose may be set by the ctdivol rather than the dlp . in this case , the exposure dose predicted value is also obtained as the ctdivol . the upper limit and predicted value for the ctdivol will be denoted by ctdivolu and ctdivolc , respectively , hereinbelow . next , at step 608 , a decision is made on whether the predicted value is greater than the upper limit . if the predicted value is greater than the upper limit , the tube current is modified at step 610 . when the tube current has been set by auto - milliampere , the modification of the tube current is performed according to the following equation : where i is the tube current set by auto - milliampere , i . e ., an unmodified tube current , and i ′ is the modified tube current . by such modification , the tube current by auto - milliampere as indicated by the solid line in fig1 is modified into one as indicated by the broken line in fig1 , for example . if the tube current i has been set without using auto - milliampere , the tube current modification is achieved as follows : the upper limit dlpu is divided by the imaged length to obtain the ctdivol , and the tube current is obtained based on the ctdivol by inverting the aforementioned algorithm . alternatively , the tube current may be obtained based on the pre - measured relationship between the ctdivol and tube current . if the upper limit is set using the ctdivol , the tube current can be obtained from the ctdivol without the division by the imaged length . the data processing apparatus 60 for conducting the processing of steps 606 - 610 is an embodiment of the modulating means of the present invention . the tube current modification may alternatively be conducted using the following equation . this facilitates the tube current modification . next , at step 612 , a scan is conducted . the scan uses the tube current modified as described above . this allows a scan to be conducted without the exposure dose exceeding the upper limit . if the predicted value does not exceed the upper limit , the unmodified tube current is used to achieve a scan without the exposure dose exceeding the upper limit . next , at step 614 , image reconstruction is conducted . a reconstructed image is displayed on the display device 68 and stored in the memory at step 616 . although the preceding description has been made on a case in which a helical scan is conducted , it will be easily recognized that the present technique is not limited to the case of a helical scan but also enables a similar effect to be obtained in conducting an axial scan . [ 0121 ] fig1 shows a block diagram of an x - ray ct apparatus , which is an embodiment of the present invention . the configuration of the apparatus represents an embodiment of the apparatus in accordance with the present invention . in fig1 , parts similar to those shown in fig1 are designated by similar reference numerals , and explanation thereof will be omitted . the present apparatus comprises a communication interface 72 . the communication interface 72 is disposed between an external communication network and the data processing apparatus 60 . the data processing apparatus 60 exchanges data with the outside via the communication interface 72 . [ 0123 ] fig1 shows a block diagram of a medical image network to which the present apparatus belongs . as shown , an image server 802 and a plurality of x - ray imaging apparatuses 812 , 814 , . . . , 81 n are connected via a communication circuit 820 to constitute a medical image network . the x - ray imaging apparatus 81 i ( i : 2 , 4 , . . . , n ) is an x - ray ct apparatus , for example . however , the x - ray imaging apparatus is not limited to the x - ray ct apparatus but may be an appropriate imaging apparatus conducting imaging using x - rays , such as an x - ray fluoroscopic imaging apparatus . the image server 802 archives images captured by each x - ray imaging apparatus 81 i ( i : 2 , 4 , . . . , n ) and associated information . the information is accessible by each x - ray imaging apparatus 81 i . if another image server 902 separate from the image server 802 is connected via the communication circuit , the x - ray imaging apparatus 81 i can access the image server 902 . the image servers 802 and 902 represent an embodiment of the server of the present invention . the present apparatus is configured to be capable of accessing the image server 802 ( or 902 or both ) to acquire historical information on a particular patient , calculating an x - ray dose to which the patient has been exposed thus far based on the information , and displaying the x - ray dose on the display device 68 . specifically , as shown in the flow chart of fig1 , upon input of patient information at step 702 , the data processing apparatus 60 acquires historical imaging data at step 704 , and calculates an exposure dose at step 706 . thus , the exposure dose over the past year is found , for example , and is displayed at step 708 . the data processing apparatus 60 for executing the processing at steps 704 and 706 is an embodiment of the calculating means of the present invention . the display device 68 for executing the display at step 708 is an embodiment of the display means of the present invention . thus , the user of the present apparatus can know the exposure dose to date for a patient . the exposure dose may be effectively used as reference data for present or future imaging to reduce the total amount of the exposure dose to the patient . although the present invention has been described with reference to the preferred embodiments hereinabove , several changes and substitutions may be made on these embodiments by those ordinarily skilled in the art to which the present invention pertains without departing from the scope of the present invention . therefore , the technical scope of the present invention is intended to encompass not only the aforementioned embodiments but all embodiments pertaining to the appended claims . many widely different embodiments of the invention may be configured without departing from the spirit and the scope of the present invention . it should be understood that the present invention is not limited to the specific embodiments described in the specification , except as defined in the appended claims .

for the purpose of enabling reduction of exposure dose , in an x - ray controlling method for an x - ray imaging apparatus for producing an image based on detected x - ray signals , an upper limit of an x - ray exposure dose to a subject to be imaged is set , and the tube current of an x - ray tube is modulated so that the exposure dose does not exceed the upper limit . the modulation of the tube current is achieved by finding an exposure dose predicted value based on an imaging protocol , and modifying a tube current set value in the imaging protocol when the predicted value exceeds the upper limit .

the collapsible containers , per se , which form a part of the invention , are not novel , and suitable containers are readily available . the only requirement is that the container be sufficiently flexible so that opposing sides readily come together when liquid is drained from the container by gravity . typically useful plastic from which the bags may be made are the polyolefins , polyvinylchlorides , ethylvinylacetates , etc . fig1 is a side view of plastic bag 10 formed by sealing the edges of planar plastic sheet 12 with the edges of planar plastic sheet 13 . handle 16 for supporting bag 10 in use is provided at the upper portion . as shown , the bag contains a liquid which has its upper surface 37 , the meniscus , in the zone between 34 and 36 . the position of the top surface of the liquid may vary depending on the volume of air space , yet its position is conventionally used to determine the apparent volume of liquid within the container . errors arising from variations in the meniscus level for a typical container having a capacity of 3 liters may be as great as several hundred milliliters of liquid or more . fig2 and 3 show the preferred embodiment of the liquid level indicating device which makes possible an increase in the accuracy and precision of measuring the volume of liquid within the bag . the preferred liquid level indicating device 31 comprises a pair of rollers 30 , 32 which are spaced apart and mounted on axle means 28 . the length of the rollers is not critical and rollers shorter than the width dimension of the bag may be used by sizing element 28 to span the width of the bag . however , the rollers are preferably long enough so that they fit easily over the collapsed bag , and most preferably have a length about equal to or slightly greater than the width of the collapsed bag . however , rollers significantly longer than the container width are also within the scope of the invention . the spacing between the rollers must be great enough so that the flattened sheets 12 , 13 in the collapsed portion of the bag will pass freely between the rollers , i . e ., the spacing between the rollers should be slightly greater than the thickness of the two layers of film which make up the bag . inasmuch as it may be necessary when assembling the bag and the liquid level indicating device for the rollers 30 , 32 to pass over a handle , such as handle 16 , when assembling the bag and the rollers , the spacing between the rollers should be sufficient to accommodate an increased thickness of plastic at that point . typically useful spacing of the rollers is from about 1 / 8 inch to about 3 / 8 inch . while the spacing between the rollers of the device 31 is not critical , it may affect the relative vertical positions of the rollers 30 , 32 and volume reading , and it may be necessary in some instances to calibrate the collapsible bag with the specific device being used , or one having appropriate dimensions . as shown in fig4 and 5 , bag 10 is provided with liquid level indicating device 31 . bag 10 is provided with handle 16 having openings 15 and 17 for suspending the container in use . also shown in fig4 and 5 are fluid outlet means 26 , inlet means 18 and 20 , and injection port means 24 and 22 . front face 12 is provided with calibrations 14 . as shown in fig5 planar sheets 12 and 13 are sufficiently close together in the zone above device 31 so that little or no liquid is present above the line of contact between rollers 32 , 30 and plastic sheets 12 and 13 , respectively . therefore , device 31 provides a positive and reproducible reference point for reading the level of liquid within the flexible container 10 . while in the preferred embodiment of device 31 as shown in the figures , the elements 30 and 32 are shown as cylindrical bodies , the invention also contemplates the use of other forms for these elements . for example , elements may be used which have a rectangular cross - section and which would not roll but instead would be capable of sliding downwardly on the outer surface of the plastic bag from the force of gravity as liquid is administered . the diameters of the elements 30 , 32 are not critical ; however , elements having diameters in the range from about 1 / 4 inch to about 1 / 2 inch are preferred . when properly sized , there is no need to force device 31 downwardly as liquid is administered , but it should just be permitted to follow the liquid surface down by virtue of gravity as liquid is removed from the bag . in the preferred apparatus , all air is excluded from the bag . however , the invention also contemplates the use of the liquid level indicating device with bags containing air in such small quantities that the accuracy of measuring the volume of liquid in the bag is not significantly reduced . having thus described the invention , the following example is given to illustrate it in more detail . an intravenous collapsible container as shown in the figures having a capacity of 3000 milliliters and dimensions when collapsed of about 91 / 2 inches by about 11 inches , was fitted with a device 31 having steel rollers made from hollow tubing which were about 101 / 8 inches long , about 3 / 8 inch in outside diameter , about 1 / 4 inch in inside diameter and having a space between them of about 5 / 32 inch . the element 28 was a steel wire about 3 / 32 inch in diameter which enters through element 30 and was bent to extend about 1 / 2 inch into each end of element 32 .

apparatus for administering medical liquids comprising a combination of a calibrated flexible bag and a liquid level indicating device for increasing the accuracy of measuring the level of liquids within the bag . the liquid level indicating device consists of a pair of spaced - apart elongated rollers which form a slot into which the upper part of the bag can be inserted . the rollers rest on the outside of the bag at about the upper level of liquid within the bag .

referring now to fig1 - 5 , the present invention features a sports team novelty system 100 for attaching to a vehicle 105 . the novelty system 100 of the present invention allows a user to display support for his / her favorite team . the novelty system 100 is designed to resemble an animal or mascot for a sports team . in some embodiments , the novelty system 100 resembles a tiger . in some embodiments , the novelty system resembles another animal or mascot such as a hornet , a bulldog , an eagle , a wildcat , a cougar , a falcon , a devil , a wolf , a trojan , or the like . the novelty system 100 comprises a first housing 110 , which is designed to resemble the front or head part of the animal or mascot ( e . g ., a tiger head , an eagle head , a hornet head ). the first housing 110 is for installing on the front 106 of a vehicle 105 . in some embodiments , the first housing 110 is attached to the front 106 of a vehicle 105 via one or more first straps 190 ( e . g ., elastic straps ), for example with first hooks 191 . the novelty system 100 of the present invention further comprises a second housing 120 designed to resemble the back portion or tail of the animal or mascot ( e . g ., tiger tail , eagle tail , hornet stinger ). the second housing 120 is for attaching to the rear 107 of a vehicle 105 . in some embodiments , the second housing 120 is attached to the rear 107 of the vehicle 105 via one or more second straps 192 ( e . g ., elastic straps ), for example with second hooks 193 . disposed inside the first housing 110 is a first microprocessor 160 . a speaker 180 is disposed on the housing 110 and is operatively connected to the first microprocessor 160 . the system 100 also comprises a first receiver operatively connected to the first microprocessor 160 . the first microprocessor 160 comprises a memory component ( e . g ., random access memory , flash memory ) programmed with various sounds , for example sounds such as sounds of animals ( e . g ., tiger roar ) and / or songs such as fight songs for teams . when the speaker 180 is activated , it emits the sound accordingly . the first microprocessor 160 and / or speaker 180 and / or first receiver are operatively connected to a power source . in some embodiments , the power source is an electrical system of the vehicle 149 or a battery . in some embodiments , the microprocessor 160 and / or speaker 180 and / or first receiver 170 are operatively connected to the electrical system of the vehicle 149 via first wiring 145 . a motor 210 and a second receiver 270 are both disposed inside the second housing 120 . the motor 210 ( when activated ) functions to cause the second housing 120 to move or rotate ( e . g ., if the second housing 120 is a tiger tail , the tiger tail can be made to wag ). such motors are well known to one of ordinary skill in the art . a second microprocessor 260 is operatively connected to the motor 210 and the second receiver . the second microprocessor 260 and / or motor 210 and / or second receiver 270 are operatively connected to a power source . in some embodiments , the power source is an electrical system of the vehicle 149 or a battery . in some embodiments , the second microprocessor 260 and / or motor 210 and / or second receiver 270 are operatively connected to the electrical system of the vehicle 149 via second wiring 148 . the novelty system 100 the present invention further comprises a remote control 310 , which allows a user to operate the system 100 while inside the vehicle 105 . disposed inside the remote control 310 are a third microprocessor 340 and a transmitter 330 , wherein the transmitter 330 is operatively connected to the third microprocessor . a plurality of control buttons 320 for operating the first housing 110 and / or the second housing 120 are disposed on the remote control 310 . for example , a first housing button may function to control the first housing 110 and a second housing button may function to control the second housing 120 . the control buttons 320 may each be operatively connected to the third microprocessor . control buttons 320 are well known to one of ordinary skill in the art . the third microprocessor is configured to receive a first input signal from the first housing button when the first housing button is pressed . upon receipt of the first input signal , the third microprocessor is configured to generate a first output command to the transmitter 330 to cause the transmitter 330 to emit a first transmitter signal to the first receiver . the first microprocessor 160 is configured to receive a first receiver input signal from the first receiver when the first receiver receives the first transmitter signal from the transmitter . upon receipt of the first receiver input signal the first microprocessor 160 is configured to generate a speaker output command to the speaker 180 to cause the speaker 180 to emit a sound . the third microprocessor is configured to receive a second input signal from the second housing button when the second housing button is pressed . upon receipt of the second input signal , the third microprocessor is configured to generate a second output command to the transmitter 330 to cause the transmitter 330 to emit a second transmitter signal to the second receiver . the second microprocessor 170 is configured to receive a second receiver input signal from the second receiver when the second receiver receives the second transmitter signal from the transmitter . upon receipt of the second receiver input signal the second microprocessor 170 is configured to generate a motor output command to the motor 210 cause the motor 210 to move the second housing 210 . in some embodiments , the third microprocessor and / or the transmitter 330 are operatively connected to a power source . in some embodiments , the power source is an electrical system of the vehicle 149 or a battery . in some embodiments , the third microprocessor and / or the transmitter 330 are operatively connected to the electrical system of the vehicle 149 via third wiring 147 . referring now to fig5 , the microprocessor stores all information for the first housing 110 ( e . g ., sounds such as tiger roar , fight songs , etc .). an audio circuit may supply the information to an amplifier ( e . g ., a 50 w amplifier ), which is then fed to the speaker . in some embodiments , the amplifier comprises volume control . the transmitter may be a uhf - fm transmitter . the 12v dc vehicle power supply may provide regulated power supply with a quick disconnect plug from the car battery . the first housing 110 and / or the second housing 120 may be constructed in a variety of sizes . in some embodiments , the first housing 110 is between about 5 to inches tall as measured from the top end 113 to the bottom end 114 . in some embodiments , the first housing 110 is between about 10 to 15 inches tall as measured from the top end 113 to the bottom end 114 . in some embodiments , the first housing 110 is more than about 15 inches tall . in some embodiments , the first housing 110 is between about 5 to 10 inches long as measured from the first side end 111 to the second side end 112 . in some embodiments , the first housing 110 is between about 10 to 15 inches long as measured from the first side end 111 to the second side end 112 . in some embodiments , the first housing 110 is more than about 15 inches long . in some embodiments , the first housing 110 is between about 5 to 10 inches wide as measured from the front end 115 to the back end 116 . in some embodiments , the first housing 110 is between about 10 to 15 inches wide as measured from the front end 115 to the back end 116 . in some embodiments , the first housing 110 is more than about 15 inches wide . as used herein , the term “ about ” refers to plus or minus 10 % of the referenced number . for example , an embodiment wherein the first housing 111 is about 10 inches wide includes a first housing 111 that is between 9 and 11 inches wide . the following the disclosures of the following u . s . patents are incorporated in their entirety by reference herein : u . s . pat . no . 6 , 197 , 390 ; u . s . pat . no . 6 , 769 , 951 ; u . s . pat . no . 6 , 037 , 679 ; u . s . pat . no . 4 , 635 , 039 ; u . s . pat . no . 6 , 288 , 633 ; u . s . pat . no . 5 , 397 , 866 ; u . s . pat . application no . 2007 / 0124972 ; u . s . pat . application no . 2006 / 0001284 . various modifications of the invention , in addition to those described herein , will be apparent to those skilled in the art from the foregoing description . such modifications are also intended to fall within the scope of the appended claims . each reference cited in the present application is incorporated herein by reference in its entirety . although there has been shown and described the preferred embodiment of the present invention , it will be readily apparent to those skilled in the art that modifications may be made thereto which do not exceed the scope of the appended claims . therefore , the scope of the invention is only to be limited by the following claims .

a novelty system for attaching to a vehicle featuring a first housing designed to resemble a front or a head of a mascot and a second housing designed to resemble a back portion or a tail of the mascot ; means of attaching the housings to the front and rear of the vehicle , respectively ; a speaker disposed in the first housing for emitting sounds such as animal sounds or fight songs ; a motor disposed in the second housing for moving or rotating the second housing ; and a remote control for controlling the first and second housings .

as shown in fig1 the preferred embodiment of a remote control system according to the present invention is comprised of a pair of transceivers : a user - directed mobile transceiver 10 ( which may or may not be in a gun - shaped housing 100 as shown ) and a fixed transceiver 50 . the transceivers 10 , 50 are used in conjunction with a microprocessor controlled gaming unit 60 , such as the genesis 16 bit mega system ® manufactured by sega corporation , and a raster scanned video display terminal 70 , such as a standard television . the gaming unit 60 , typically under the control of a gaming program contained in a cd rom 61 , or other such storage means , generates a video signal 62 which causes the television 70 to display sequential game images that may be varied as the game progresses and in response to various user inputs . during operation , the user may aim the mobile transceiver 10 at a target area 74 on the television 70 , whereby image data 73 contained within the target area 74 is focused onto a sensor device ( described later ) contained in the mobile transceiver 10 and then communicated to the gaming unit 60 via the fixed transceiver 50 . the gaming unit 60 is generally connected to the fixed transceiver 50 with a hard link 52 . the fixed transceiver 50 provides a two - way wireless communications link 54 between the gaming unit 60 and the mobile transceiver 10 , whereby the gaming unit 60 obtains the image data 73 from which it can determine the target location 74 . in response to the image data 73 , the gaming unit 60 locates the target area 74 and , if desired , modifies the video signal 62 to generate a cursor 71 at the target location 74 . fig2 is a schematic block diagram of the mobile transceiver 10 shown in fig1 . fig6 is an actual schematic of the presently preferred circuitry for implementing the mobile transceiver 10 . to the extent possible , portions of the circuitry shown in fig6 have been identified as corresponding to the functional blocks of fig2 . in view of the functional diagram of fig2 it is not believed necessary to include a blow - by - blow signal level description of the circuit shown in fig6 . as shown in fig2 the circuitry of the mobile transceiver 10 may be conceptually divided into a intermediate ir transmit section 30 and two signal generating sections : an upper target focusing section 20 and an ir receive section 40 . the upper target focusing section 20 and the lower ir receive section 40 are each connected to the ir transmit section through an or gate 31 . a pulse received at either input to the or gate 31 causes a corresponding pulse to be issued at an output 31a . the output 31a of the or gate 31 is connected to a one - shot 32 which then generates a 10 μsec pulse on its output 32a . a driver 33 illuminates a pair of ir leds 34 , 35 upon receipt of each one - shot output 32a pulse . the target focusing section 20 monitors the image data 73 contained with the target area 74 of the television 70 . the image data 73 is focused through a lens 24 onto a phototransistor 22 which is sensitive to visible light . thus , every time a horizontal scan line 76 passes through the target area 74 , the phototransistor 22 generates a pulse which is amplified by an amplifier 25 and output as an amplified pulse signal 25a . a circuit 26 generates a reference voltage which is provided as a reference signal 26a . a comparator 27 compares the amplified pulse signal 25a with the reference signal 26a and , whenever the former exceeds the latter , outputs a horizontal scan detection signal 27a to the ir transmit section 30 . the target focusing section 20 features an optical biasing led 21 which , in the preferred embodiment , is physically located immediately adjacent to the phototransistor 22 . the gain of the phototransistor 22 is very low at low collector currents ( low light ). the gain generally increases as the collector current increases . the use of the optical biasing led 21 allows for the two - terminal phototransistor 22 instead of a standard three - terminal phototransistor having an electrical base drive input . the latter device is undesirable because an attempt to increase gain by biasing the base of the transistor may actually reduce gain . in particular , the resistive network used to electrically bias the device may act as a low impedance shunt for any photocurrent generated by the device . it has been discovered that a sensitivity increase of 10 : 1 can be obtained with the optical biasing led 21 , as compared to a sensor in darkness . a bias driver 23 provides the necessary current to drive the biasing led 21 at a current which makes it sufficiently bright . because the mobile transceiver is powered by batteries ( not shown ), the drive current provided by the bias driver 23 must be a compromise between brightness and reasonable battery life . at the present time , the preferred bias led 21 is red in color and is driven at 10 ma . it is supposed that amber or orange leds may be preferred over red because of their generally higher quantum efficiencies . the ir receive section 40 outputs button press data 45a to the ir transmit section 30 every time an ir pulse is sensed by an ir sensitive diode 41 . an output of the diode 41 is enhanced with an amplifier 42 . thus , in response to the diode &# 39 ; s receipt of an ir pulse , the amplifier 42 provides an amplified pulse signal 42a . a circuit 43 generates a reference voltage which is provided as a reference signal 43a . a comparator 44 compares the amplified pulse signal 42a with the reference signal 43a and , whenever the former exceeds the latter , outputs an ir pulse detection signal 44a to an enable input of a pulse generator 45 . each time the pulse generator 45 receives a pulse at its enable input , it provides the button - press data 45a to the ir transmit section 30 . in the preferred embodiment , the ir receive section 40 operates with four switches sw1 , sw2 , sw3 , and sw4 . the button - press data 45a is preferably comprised of a variable string of pulses , the number of pulses based on which one or more the switches sw1 , sw2 , sw3 , sw4 was depressed at the time an ir pulse was sensed by the diode 41 . the following table identifies a preferred encoding system wherein sw1 is identified by one pulse , sw2 is identified by two pulses , sw3 is identified by four pulses , and sw4 is identified by eight pulses . if two or more of the switches was pressed , then the number of pulses generated by the pulse generator 45 is equal to the sum of the foregoing pulses . on the basis of this encoding system , each series of between 0 and 15 pulses uniquely identifies each possible combination of the four switches sw1 , sw2 , sw3 , sw4 as follows : ______________________________________no . of pulses switches depressed______________________________________0 none1 sw12 sw23 sw2 sw14 sw35 sw2 swl6 sw3 sw27 sw3 sw2 sw18 sw49 sw4 sw110 sw4 sw211 sw4 sw2 sw112 sw4 sw313 sw4 sw3 sw114 sw4 sw3 sw215 sw4 sw3 sw2 sw1______________________________________ fig3 depicts a functional block diagram of the fixed transceiver 50 and fig7 is an actual schematic of the presently preferred circuitry for implementing the same . to the extent possible , portions of the circuitry shown in fig7 have been identified as corresponding to the functional blocks of fig3 . in view of the functional diagram of fig3 it is not believed necessary to include a blow - by - blow signal level description of the circuit shown in fig7 . like schematic diagram of the mobile transceiver 10 shown in fig2 the diagram of the fixed transceiver 50 shown in fig3 may also be conceptually divided , but in this case into only two sections : an ir transmit section 80 and an ir receive section 90 . as shown in fig1 the fixed transceiver 50 is connected to the gaming unit 60 via the connector plug p1 and the cable 52 . the gaming unit 60 , under the control of appropriate software , receives data from the fixed transceiver 50 and controls its operation . the fixed transceiver &# 39 ; s ir transmit and receive sections 80 , 90 respectively &# 34 ; talk &# 34 ; and &# 34 ; listen &# 34 ; to the conversely named ir receive and transmit sections 40 , 30 of the mobile transceiver . in the preferred embodiment , both transmission protocols are relatively simple , i . e . pulses of fixed duration . the gaming unit 60 controls the operation of the fixed transceiver 50 with a variable length reset signal rst . as shown in fig3 ., the reset signal rst is provided to a counter 92 and a latch 93 in the ir receive section 90 and to a pulse length detector 82 in the ir transmit section 80 . in the preferred embodiment , the reset signal rst has only two different lengths : a counter reset signal which is between 2 - 3 μsec long and a main reset signal which is about 10 μsec long . the counter reset signal is issued after the counter data ( switch data ) is read at the end of the vertical blanking interval . the main reset signal is issued once at the beginning of each vertical retrace , i . e . 60 times per second at the beginning of each television frame . during the vertical blanking interval , no scan lines are present . thus , the receiver may differentiate between the data from the button encoder and the visible scan line pulses . switch data is generated during the vertical blanking interval and , therefore , there can be no scan lines present . to do this , a sync signal is provided at the beginning of the vertical retrace interval , and the mobile transceiver sends switch data . the game unit then reads the counter of the fixed transceiver at the end of the vertical interval , before any scan lines are generated . the game unit then resets the counter ( without generating an ir sync pulse ) and waits for scan lines to appear in the field of view . the ir receive section 90 is comprised of an ir sensitive led 91 which is connected across an amplifier 94 . in response to the detection of an ir pulse by the diode 91 , the amplifier 94 provides an amplified pulse signal 94a . a comparator 44 compares the amplified pulse signal 94a with a reference signal 95a provided by a circuit 95 . the output 96a of the comparator is provided as a clock clk input to a counter 92 configured as an up counter . thus , for each horizontal scan line 76 crossing the target area 71 and the corresponding ir pulse detected by diode 91 , the counter 92 counts up , with its outputs q1 , q2 , q3 , q4 going successively from 0000 to 1111 . of all the scan lines which cross the target area , the central most scan line will provide the longest and highest intensity pulse , which will provide the least amount of timing variation from frame to frame . on average , when the preferred mobile transceiver 10 is about ten feet away from a 21 &# 34 ; television , the target area 74 encompasses about 20 scan lines . thus , when the count reaches 8 , i . e . when q4 goes high to provide binary 8 ( 1000 ), then latch 93 is set and provides a scan line detection signal to the gaming unit 60 until it receives the reset signal rst . in order to accommodate other screen sites , other counts may be used . in the ir transmit section 80 , the pulse length detector 82 ignores the shorter counter reset signal , but , in response to a longer main reset signal outputs a reset detection signal 82a to a driver 84 , which in turn pulses a pair of ir leds 85 , 86 . the ir pulse may be referred to as a vertical retrace pulse in that the ir leds 85 , 86 are triggered at the beginning of each vertical retrace . the transmission of the vertical retrace pulse is the signal for the mobile transceiver 10 to send button - press data , if any . the communication between the two transceivers can be summarized as follows . the mobile transceiver continuously transmits image position data comprised of a single pulse each time a horizontal scan line 76 passes through the target area 74 and , immediately after having received a vertical retrace pulse from the fixed transceiver ( time phase information ), transmits button - press data comprised of between 0 and 15 pulses depending on the number of buttons currently pressed by the user , if any . fig4 and 5 are respectively a perspective view and an exploded view of a preferred multipiece gun - shaped housing 100 for the mobile transceiver 10 . as best shown in fig5 the housing 100 is comprised of three main components , separable from one another : a main body 110 , a stalk 130 , and a sighting system 150 . the main body 110 is comprised of a tube portion 112 , a u - shaped grip 114 having a front member 118 and a rear member 116 . the main body 110 actually houses the circuitry for the mobile transceiver shown in fig2 . the lens 24 which focuses the target area 74 onto the phototransistor 22 is located at a front end of the tube portion 112 as shown in fig5 . as best shown in fig4 the four switches sw1 , sw2 , sw3 , sw4 are exposed on the exterior of the main body 110 . one switch sw2 serves as the &# 34 ; fire &# 34 ; switch and is positioned on the rear member 116 for activation by the user &# 39 ; s index finger . the other three switches sw1 , sw3 , sw4 are located on the front member 118 for activation by the fingers of the user &# 39 ; s other hand . thus , the mobile transceiver 10 and housing 100 can serve not only as a targeting device (&# 34 ; fire &# 34 ; button only ), but also as a multibutton controller , with or without a visible cursor . the tube portion 112 of the preferred main body 110 also includes two adjacent pairs of slots , an upper pair of slots 115 , and a lower pair of slots 117 . as suggested by the dashed lines in fig5 the upper pair of slots 115 receive the sighting system 150 and the lower pair of slots receive the stalk 130 . the stalk 130 is comprised of a body portion 132 and a pair of opposed inwardly - extending tabs 134 which surround a hollow portion 136 . the sighting system is likewise comprised of a pair of opposed inwardly - extending tabs 154 which surround a hollow portion 156 . the inwardly extending tabs 154 , 134 respectively engage the upper and lower pairs of slots 115 , 117 at the rear of the main body 110 . by this novel arrangement , the user is provided with a versatile easily - modified housing 100 for the mobile transceiver 10 . with all three components 110 , 130 , 150 connected to one another , the user has a rifle - like housing 100 with a sighting system 130 . under certain conditions , the user may prefer to use just the main body 110 and the stalk 130 . under even other conditions , the user may use just the main body 110 in a pistol - like fashion . the sighting system 150 is further comprised of a pair of viewing tubes 152 which appear like binoculars . in the preferred embodiment , the viewing tubes 152 do not contain any optics , but simply are configured with an interior transverse wall having a small hole through which the user may look . a rotatable member 153 having a hollow sight tube 158 is provided at a front end of a member 157 which extends forward of the viewing tubes 152 . the hollow sight tube 158 may be rotated in front of a desired one of the viewing tubes 152 . the user would ordinarily rotated the hollow sight tube 158 in front of the viewing tube 152 serving the user &# 39 ; s strongest eye ( i . e , the dominant eye ). those skilled in the art will appreciate that various adaptations and modifications of the just - described preferred embodiment can be configured without departing from the scope and spirit of the invention . therefore , it is to be understood that , within the scope of the appended claims , the invention may be practiced other than as specifically described herein .

the present invention is a remote control targeting and control system for use with a standard raster scanned television and an associated gaming unit . the remote control system includes a mobile transceiver and a fixed transceiver which communicates with the gaming system . the system implements a simple communication system which allows the mobile transceiver to ordinarily transmit position data and to periodically transmit button press data . the mobile transceiver includes an electro - optic detector for observing when a scan line crosses a target area . each time a scan line is detected , the mobile transceiver transmits a pulse to the fixed unit , based upon which the gaming unit translate the pulse into position data based on the time of its detection . the button press data is transmitted synchronously with the raster scanned television in that the fixed transceiver is caused to transmit a single pulse during each vertical retrace of the television . the preferred remote control system is contained in a gun - shaped main unit of a multipiece housing . a stalk and a sighting system having viewing tubes may be optionally attached to the main unit . the sighting system is comprised of a pair of viewing tubes and a rotatable sighting tube which may be place in front of either sighting tube .

this invention will be best understood by referring to the drawings . looking now at the blood taking syringe of fig1 , note the syringe housing ( 4 ) having first and second chambers , ( 4 a ) and 4 ( b ), respectively , with the first chamber ( 4 a ) adapted to receive a blood taking vacuum vial ( 5 ) and with the second chamber ( 4 b ) designed to hold a spring loaded , double ended needle ( 1 a ) within a needle holder head ( 1 b ) of the hypodermic needle head assembly ( 1 ). the double ended needle ( 1 a ) is mounted within a needle head assembly ( 1 ) and extends axially therethrough , allowing the distal end of the needle to be inserted into a vein of a blood giver while the other end punctures one end of a vacuum vial ( 5 ) upon inserting the vial into chamber ( 4 a ) and inserting moderate pressure thereto . as is well known in the art , the vacuum within the vacuum vial ( 5 ) then draws blood from the vein of the patient and fills the vial accordingly . any desired number of vials of blood may be drawn from the patient before initiating the withdrawal of the needle from the patient and releasing the needle head assembly ( 1 ) for storage within chamber ( 4 a ) of syringe housing ( 4 ). a compressed spring ( 3 ) tends to force the needle assembly ( 1 ) into the inside of the upper chamber ( 4 a ) of the syringe housing ( 4 ), however , spring ( 3 ) is prevented from driving the head assembly ( 1 ) into chamber ( 4 a ) by an annular hold - down ring ( 2 ), which can be fabricated of hard plastic or vinyl or any other material strong enough to withstand the force of the compressed spring ( 3 ) and apply a counter force to the needle assembly ( 1 ) and hold the needle assembly in a state of equilibrium from the force of the compressed spring ( 3 ). retraction of the needle head assembly ( 1 ) may be effected , upon demand , by depressing the release pin ( 7 ), whereupon the end of pin ( 7 ) is forced against the edge of the hold - down ring ( 2 ), forcing the ring into the center of the syringe housing ( 4 ). note that a syringe projection , having a slot ( 4 c ) therein , extends entirely around the periphery of the syringe housing ( 4 ) at the intersection of the syringe housing chambers ( 4 a ) and ( 4 b ) and that the slot is of such a width and depth that the thickness of the annular ring ( 2 ) readily fits therein . the annular ring ( 2 ) has a hole in the center thereof sufficiently large to allow the needle assembly to unobstructively pass therethrough upon alignment of the axis of annular ring ( 2 ) and the central axis of the syringe housing ( 4 ), however , when the was of the annular ring ( 2 ) and the axis of the syringe ( 4 ) are not coincident , the annular ring , being off - center , functions to restrain the needle assembly ( 1 ) by stopping the upward movement of the needle assembly ( 1 ) with one edge of the annular ring ( 2 ) being in contact with needle holder ( 1 b ). the centerline axis of the annular ring is normally offset from the central axis of the syringe housing ( 4 ), thusly restraining the needle assembly ( 1 ) until activation of the release pin ( 7 ). prior to activating the release pin ( 7 ), the safety cap ( 9 ) is attached to the syringe housing , which is here shown as a screw - on type , but may be of any design that would securely seal the cap when applied to the container . the safety cap ( 9 ) is shown as twisted onto the guide ( 10 ) and may be similar to a push down , commercial safety cap used on safe liquid medicine bottles for protecting children . the safety cap seal ( 12 ) seals the top of chamber ( 4 a ) upon placing the safety cap ( 9 ) on the syringe housing and accordingly seals in any fluid remaining in chamber ( 4 a ) upon removal of the vacuum vial ( 5 ), as well as sealing chamber ( 4 a ) for accepting the needle assembly upon release of the needle restraining means . safety cap ( 11 ) is only one example , among many , that may be used to effect the sealing of the needle within the syringe . broken rings ( 13 ) on the cap screw are mated with ring guide ( 10 ) on the housing . when the cap is screwed onto the housing , it seals that end of the housing , eliminating the possibility of the inadvertent removal of the cap and spilling contaminated fluid . upon activation of the needle release means the compressed spring ( 3 ) forces the needle head assembly ( 1 ) into chamber ( 4 a ) of the syringe for storage . a push - in motion applied to the push tab ( 7 ) releases the holding ring ( 2 ) which can be performed easily with one hand . the spring then would be free to expand and force the needle head assembly into the container holder . the inner cylinder ( 8 ) provides a guide for vacuum vial ( 5 ), but is not an essential part of the syringe . if desired , the inner cylinder ( 8 ) may be molded into the syringe upon manufacture or may be a simple sleeve inserted within the syringe housing . once the needle assembly is retracted into the syringe housing chamber ( 4 a ), it cannot be reused . when the cap is screwed onto the housing , it seals that end of the housing , eliminating the possibility of the inadvertent removal of the cap and spilling contaminated fluid . seal ( 14 ) is a self - sealing seal that immediately seals the hole left by the needle after retraction . this self - sealing material seals the lower end of the syringe housing , which prevents any inside contaminated fluid from leaking to the outside and prevents the retracted needle from protruding back through the hole left by retraction of the needle ( 1 a ). cavity ( 15 ) captures any droplets of blood that might be wiped off the needle ( 1 a ) by the seal ( 14 ) during retraction , however , an additional cap ( not shown ), similar to that of cap ( 9 ), may be used on the outermost end of the second chamber ( 4 b ) of the syringe housing ( 4 ), in the event that one desires to be assured that the spent needle and other body fluids are absolutely encapsulated within the housing upon retraction of the needle assembly . the two end caps ( 9 ) may be attached to the syringe upon manufacture and removed prior to use , in order to protect the needle from any potential environmental contamination . upon attaching safety caps ( 9 ) on the outermost ends of chambers ( 4 a ) and ( 4 b ) and upon retraction of the needle assembly ( 1 ) into chamber ( 4 a ) of the syringe housing ( 4 ), the needle head assembly and any contaminated fluids within the housing of the syringe ( 4 ) are encapsulated within the syringe housing ( 4 ), thereby rendering the syringe housing ( 4 ) a safe container for users and handlers who may come in contact with the used syringe . fig2 shows the blood - taking device of fig1 in its retracted state . safety cap ( 9 ) has been twisted onto the top of the container , effectively sealing and securing the top end of the syringe . push tab ( 7 ) has been activated , forcing the annular ring ( 2 ) into central alignment with the axis of the of the syringe housing and thereby releasing the spring biased needle assembly ( 1 ) and forcing the needle assembly ( 1 ) into chamber ( 4 a ) of the syringe housing ( 4 ). the needle assembly ( 1 ) and spring ( 3 ) have been retracted . the needle seal ( 14 ) has automatically closed , sealing the bottom of the container . the container cannot be used again , thus the needle is inaccessible and the container is completely sealed , thereby rendering the used syringe ( 4 ) a safe container for the spent needle and fluids . while this invention has been described in terms of a preferred embodiment , those skilled in the art will recognize that the invention can be practiced with modification within the spirit and scope of the appended claims .

a blood - taking syringe having a retractable needle , releasable upon demand , having a needle assembly , under tension from a compressed spring , with a needle holding head supporting a double ended needle axially aligned with the central axis of the syringe , which is held in equilibrium by a releasable annular ring interposed within a slot on the inside of the syringe housing . a release pin extends through the side of the syringe at the point where the annular ring is located and is disposed such that upon depressing the pin the ring is dislodged from the slot in the syringe housing , releasing the needle assembly to be forced into the syringe for safe storage .

as illustrated in fig2 , an x - ray detectable bioabsorbable bone screw 1 comprises a screw member 12 and a cap member 11 . the screw member 12 has a cylindrical shape . the cap member 11 is provided on an end of the screw member 12 and extended outwardly from the screw member 12 . the cap member 11 and the screw member 12 are formed integrally , and the cap member 11 and the screw member 12 are made of polylactice acid and iron oxide nanoparticles . a nanoparticle is defined as a microscopicparticle with at least one dimension less than 100 nm . preferably , the sizes of the iron oxide nanoparticles are less than 50 nanometer in size . as shown in the flow diagram of fig1 , the method of manufacturing the x - ray detectable bioabsorbable bone screw is described according to steps s 1 to s 8 . the method of manufacturing the bone screw comprises steps of : providing an injection molding device , a die , polylactice acid and iron oxide nanoparticles , wherein the die is provided with a bone screw mold , and the injection molding device is connected to the die ; mixing the polylactice acid and the iron oxide nanoparticles to form a raw material ; granulating the raw material to form a raw granule ; providing the injection molding device with the raw granule ; pressurizing and heating the raw granule with the injection molding device until the raw granule reach a predetermined state ; providing the die with the raw granule of in the predetermined state to form a bone screw shaped by the bone screw mold . the embodiment of the present invention will be described in detail with reference to the appended drawings hereinafter in order to make the objects , characteristics and advantages of the present invention more clear and comprehensible . injection molding method , a conventional plastic processing method , is employed to manufacture a bone screw model . pure pla granules and iron oxide nanoparticles are mixed to form a raw material , and the raw material is granulated to obtain a pla raw granule containing 20 wt % iron oxide nanoparticles as a raw granule . the raw granule is pressurized and heated by the injection molding device until the raw granule reaches a predetermined state . the die is filled with the raw granule in the predetermined state rapidly , and the pressure is kept constant until the raw material is cooled . after the raw material is cooled , the raw material is demolded to form a bone screw model which is shaped by the bone screw mold . the bone screw model is manufactured by an injection molding process . the bone screw model can be used to make bone screws of specific size . more specifically , the bone screw model can be used to manufacture , by injection molding process , a bone screw with a length of 11 - 20 mm in an axial direction , and a diameter of 2 . 5 - 3 . 6 mm . in this preparation example , as shown in fig3 , the bone screw model is used to manufacture bone screws with a length of 2 . 5 mm in the axial direction , and a diameter of 3 . 1 mm . however , the present invention is not limited to this . furthermore , in order to make the present invention applicable to various parts of the body to be treated and various bone diseases , other bioabsorbable material , such as hydroxyapatite , β - tricalcium phosphate and calcium polyphosphate , is selectively added into the pla raw material . in addition , pla can be substituted by materials as follows : polyglycolic acid ( pga ), polycaprolactone ( pcl ), polyethylene glycol ( peg ), lactic acid - glycol copolymer , polydioxanone ( pdo ) and / or chitin . the iron oxide nanoparticles include fe 2 o 3 , fe 3 o 4 and feo . iron oxide nanoparticles can be substituted by materials as follows : nano - sized oxides of iron , cobalt , nickel and / or chromium , such as fe 2 coo 4 , nio and cro 2 . besides the injection molding process , the bone screw can be manufactured by a 3d printing process . the bone screw model is rod - shaped and can be applied to a 3d printing process , in which the bone screw model is used to manufacture a bone screw by a 3d printing process . as mentioned above , pure pla raw material and iron oxide nanoparticles are mixed to form a raw material , and the raw material is granulated to obtain a pla raw granule containing 20 wt % iron oxide nanoparticles as a raw granule . the raw granule is pressurized and heated by the injection molding device until the raw granule reaches a predetermined state . the die is filled with the raw granule in the predetermined state rapidly , and the pressure is kept constant until the raw material is cooled . after the raw material is cooled , the raw material is demolded to form a bone screw model with a length in an axial direction of is 20 mm and a diameter of 1 . 65 mm . the bone screw model , which is rod - shaped , can be applied to an additive manufacturing device . a bone screw of 16 mm in length and 3 . 1 mm is formed by utilizing 3d printing process ( as shown in fig4 ) with the bone screw model being used as a 3d printing model . furthermore , in order to make the present invention applicable to various parts of the body to be treated and various bone diseases , other bioabsorbable material , such as hydroxyapatite , β - tricalcium phosphate and calcium polyphosphate , is selectively added into the pla raw material . in addition , pla can be substituted by materials as follows : polyglycolic acid ( pga ), polycaprolactone ( pcl ), polyethylene glycol ( peg ), lactic acid - glycol copolymer , polydioxanone ( pdo ) and / or chitin . the iron oxide nanoparticles include fe 2 o 3 , fe 3 o 4 and feo . iron oxide nanoparticles can be substituted by materials as follows : nano - sized oxides of iron , cobalt , nickel and / or chromium , such as fe 2 coo 4 , nio and cro 2 . in order to prove that the cap member 11 and the screw member 12 containing 0 . 5 to 40 weight percent of iron oxide nanoparticles is able to be manufactured and is radiographable under x - ray exposure , polylactice acid and iron oxide nanoparticles are mixed in different mix proportions by weight , producing fe 3 o 4 / pla samples at different weight ratios : 0 wt %, 20 wt %, 30 wt % and 40 wt %. a plurality of i - shaped specimens for tensile strength testing are manufactured with the fe 3 o 4 / pla samples by an injection molding process according to the astm ( american society for testing and materials ) d638 type v testing standard . the i - shaped specimens have the following specifications : a width of 2 . 6 mm , a thickness of 4 . 2 mm and a parallel length of 20 mm during the tensile strength testing , the i - shaped specimens manufactured by the injection molding process are provided on a clamping member of the tensile strength testing device , and a set the strain rate to be 2 mm / min to measure and record the yield strength of the i - shaped specimens . the yield strength of the i - shaped specimens is shown in table i . the bone screw is immersed in sterile water and oscillated by an ultrasonic oscillator for 10 minutes , and then immersed in 75 % alcohol as a cleaning step . the bone screw is sterilized by γ - ray , after which the preoperative preparation for an animal experiment is completed . a new zealand white rabbit is anesthetized by receiving a subcutaneous cervical injection ; an implantation area at the front hind leg of the new zealand white rabbit was shaved ; a position in the implantation area to be cut is locally anesthetized with lidocaine ; subcutaneous layer and muscular layer are cut open with surgical scalpel blades no . 15 from a marked point in the middle of the two joints at both ends of the femur along a front side of the femur in a long axial direction ; and the periosteum above the femur is lifted to expose femur . a proximal portion of the femoral diaphysis that is away from the articular pan is determined to be implanted . the bone tissue of the part to be implanted is drilled by a drilling machine and is rinsed with physiological saline , and the physiological saline is extracted . after the drilling is completed , the bone screw is implanted , and the subcutaneous layer and the muscular layer are sutured with 5 - 0 absorbable suture , and then epidermis is sutured with suture . the rabbit is sacrificed 4 weeks after the surgery . the femur containing the bone screw is resected and immersed in a fixative solution containing 10 % formalin . then , the femur sample containing the bone screw is dehydrated through multiple processes , and then is wrapped with paraffin and sliced to form a paraffin section . the paraffin section is stained by hematoxylin - eosin staining method and then a section slide of the paraffin section is prepared using mounting media . then , the femur sample containing the bone screw is scanned by a slice - scanner , and an image file of the section slide obtained therefrom is observed . after the bone screw of the present invention is degraded , the substance released from degradation is shown as the black parts in fig5 . the substance does not initiate an inflammatory response within body and can be covered by bone tissue , on which bone cells can grow . it is thus indicated that the bone screw of the present invention is biocompatible . the bone screw is immersed in sterile water and oscillated by an ultrasonic oscillator for 10 minutes , and then immersed in 75 % alcohol as a cleaning step . the bone screw is sterilized by γ - ray , after which the preoperative preparation for an animal experiment is completed . a new zealand white rabbit is anesthetized by receiving a subcutaneous cervical injection ; an implantation area at the front hind leg of the new zealand white rabbit was shaved ; a position in the implantation area to be cut is locally anesthetized with lidocaine ; subcutaneous layer and muscular layer are cut open with surgical scalpel blades no . 15 from a marked point in the middle of the two joints at both ends of the femur along a front side of the femur in a long axial direction ; and the periosteum above the femur is lifted to expose femur . a proximal portion of the femoral diaphysis that is away from the articular pan is determined to be implanted . the bone tissue of the part to be implanted is drilled by a drilling machine and is rinsed with physiological saline , and the physiological saline is extracted . after the drilling is completed , the bone screw is implanted , and the subcutaneous layer and the muscular layer are sutured with 5 - 0 absorbable suture , and then epidermis is sutured with suture . the rabbit is sacrificed 4 weeks after the surgery . the femur containing the bone screw is resected and immersed in a fixative solution containing 10 % formalin . the femur section is scanned by a micro computed tomography ( micro ct ) scanner for observation . the parts of the femur sample to be observed , including the bone screw and the bone tissue around the bone screw , are circled using a built - in instructional tool of the micro computed tomography to calculate the volume of new bone . as shown in fig6 a , the bone screw made of pure pla in bone is radiopaque when being exposed to x - ray radiation . as shown in fig6 b , the bone screw of the present invention is clearly shown in the bone when being exposed to x - ray radiation . it is thus indicated that the bone screw of the present invention is radiographable under x - ray exposure . as shown in fig7 , a result of a micro - ct based quantitative analysis shows a greater amount of new bone around the implanted x - ray detectable and bioabsorbable bone screw prepared according to the above - mentioned preparation examples compared with the amount of new bone around an implanted conventional bone screw . according to the above examination result , the bioabsorbable bone screw containing pla and iron oxide nanoparticles of the present invention is radiographable , and thus can be used for radiographic inspection . therefore , the bioabsorbable bone screw containing pla and iron oxide nanoparticles of the present invention can be applied to orthopedics , neurosurgery and plastic surgery to improve the positioning of the implanted bone screw and to achieve the effect of accelerating bone healing . however , the present invention is not limited to the above mentioned advantages . the above description should be considered as only the preferred embodiments of the present invention , and the scope of the embodiment of present invention is not limited thereto . various equivalents and modifications without departing from the appended claims and the description of present invention are included in the scope of the present invention .

a x - ray detectable bioabsorbable bone screw comprises a light - emitting element , a light - sensing element , a transparent inner encapsulant body , an outer covering body , and two conductive frames on which . an optically reflective surface is in contact and formed between the dome enclosing portion of the transparent inner encapsulant body and the outer encapsulant body . a portion of the light emitted by the light - emitting element is reflected to the light - sensing element through the optically reflective surface , and the other portion of the light emitted from the light - emitting element is directly emitting to the light - sensing element through the transparent inner encapsulant body . the present invention applies the optically reflective surface to minimize the overlapping area between the two conductive frames , and reduces the capacitance value , and increases the cmrr in a manner that the photo coupler of the present invention is able to meet the standard of electrical characteristics as required .

in accordance with the present invention , instead of obtaining a smear from a part of the body where it is suspected that micro - organisms are present and placing the smear on a culture medium which is then incubated , the culture medium is placed directly in the body cavity at the area where the presence of the micro - organisms is suspected , and after a time normally required for growth of the micro - organisms , if any , the device of the invention is removed from the body cavity and examined so as to diagnose whether or not there are indeed micro - organisms present at the body cavity . in this way it becomes unnecessary to use incubators , to transfer a smear to a culture medium , and the diagnosis is made as rapidly as possible and as accurately as possible since the growth of the micro - organisms , if any , takes place directly in the body under precisely the same conditions which prevail in the body of the patient . thus , referring to fig1 there is schematically illustrated therein a part 10 of the body of a human being , this part 10 having the body cavity 12 . this schematically illustrated body cavity 12 may be any cavity of the body such as the mouth , the nose , a sinus passage , the ear , the anal canal , the vagina , the urethra , the uterus , etc . assuming that a physician suspects a given area of the body as being infected by a micro - organism , the physician will simply introduce into the body cavity where the presence of infection is suspected a device 14 according to the present invention . this device 14 includes a culture medium 16 ( fig2 ) which may be any known culture medium which will promote the growth of micro - organisms if the latter are present . since such micro - organisms normally are not capable of retaining a given shape and size , the culture medium 16 is carried by a carrier means which includes a body 18 of compressed filamentary material and an outer frame 20 which carries the compressed body 18 of filamentary material . this body of filamentary material 18 may be compressed cotton fibers , fine gold wire filament , or any fine plastic monofilaments such as nylon or polyester filaments which when compressed form a large surface area having a large number of interstices in which the nutrient or culture medium 16 will become located . the frame 20 forms a shell which surrounds the body of compressed filamentary material 18 to retain the latter in its compressed condition . this frame 20 in the example of fig2 includes a pair of receptacles portions 20a and 20b joined together at their mating flanges 22 and releasably held together for example by gluing these flanges to each other if desired . the walls of the shell 20a and 20b are formed with a large number of openings 24 through which the micro - organisms will have free access to the culture medium 16 . in the embodiment of fig3 and 4 , in order to expose as large an area as possible of the culture medium , the compressed body 18 of filamentary material which supports and carries directly the culture medium 16 is situated within and surrounded by a frame 26 made up of a number of intersecting ribs 28 which provide the relatively large open spaces indicated in fig3 and 4 so that the culture medium is almost entirely exposed at the outer surface of the compressed body of filamentary material 18 . according to the embodiment of fig5 which is particularly adapted for use in testing for gonorrhea or vaginitis , the device of the invention such as that of fig2 or that of fig3 is incorporated into a tampon 30 of the type which is normally inserted into the vagina during the mentrual cycle , this tampon 30 having the string 32 which enables it to be conveniently removed . thus , fig5 shows a pair of the devices 14 incorporated into the tampon 30 which may then be conveniently introduced into the vagina to remain therein for a period of time sufficient to permit micro - organisms if they are present to grow at the culture medium in the devices 14 . then the tampon 30 is removed and the physician may examine the devices 14 to determine whether or not micro - organisms are present . with respect to the devices of fig2 and 3 , the outer frame such as the frame 20 of fig2 or the frame 26 of fig3 may be made of any metal or plastic compatible with the human body . the compressed body of filamentary material may be made of any of the filamentary materials referred to above or combinations thereof , or in fact any filamentary material compatible with the human body . the following constitutes a diagnostic medium for determining the presence of candida albicans in vaginal infection . this basic medium is made antibacterial by the addition thereto of neomycin sulfate . yeast extract 0 . 1 % glycine 1 . 0 % dextrose 1 . 0 % bismuth sulfite indicator 0 . 8 % neomycin sulfate 0 . 0002 % agar 2 . 0 % water q . s . 100 . 0 % when c . albicans is present black colonies appear , while any other fungus will not give this type of growth . bacteria are inhibited by both the neomycin sulfate and bismuth sulfite indicator . ______________________________________beef heart infusion 500 gm . tryptose 10 gm . sodium chloride 5 gm . agar 20 gm . water , q . s . 1000 ml . ______________________________________ peptone 10 gm . lactose 5 gm . sucrose 5 gm . dipotassium phosphate 2 gm . agar 20 gm . eosin y 0 . 4 gm . methylene blue . 065 gm . water , q . s . 1000 ml . this shows a green metallic sheet with e . coli . 110 media______________________________________yeast extract 2 . 5 gm . tryptose 10 . gm . gelatin 30 gm . lactose 2 gm . d - mannitol 10 gm . sodium chloride 75 gm . dipotassium phosphate 5 gm . agar 20 gm . water , q . s . 1000 ml . ______________________________________ urea - agar base______________________________________peptone 1 gm . dextrose 1 gm . sodium chloride 5 gm . monopotassium phosphate 2 gm . urea 20 gm . phenol red . 012 gm . agar 20 gm . water , q . s . 1000 ml . ______________________________________ thus , with any suitable culture medium , such as those in the examples above , it is possible for the physician utilizing the device of the present invention , simply to introduce the device directly into the body cavity where it is suspected that the micro - organisms are present . the device is permitted to remain in the body cavity for the period of time required for growth of the micro - organism , if indeed it is present , and thereafter the physician will remove the device of the invention and will examine it so as to determine whether or not micro - organisms are indeed present . since the device of the invention is incorporated directly into the body cavity to remain there at body temperature and under the exact conditions prevailing in the body , ideal conditions for growth of micro - organisms , if they are present , are used , and thus a far more accurate diagnosis is possible with the present invention than with conventional techniques and apparatus . in addition , any physician can readily practice the method of the invention without requiring the use of special equipment , laboratories , or the like , so that not only is there a great convenience and high accuracy in the diagnosis , but in addition there is a minimum of delay in making the diagnosis . in any of the above devices , the culture medium may be divided into several sections for indicating , respectively , the presence of different micro - organisms , so that a broad spectrum type of differential devices is utilized for simultaneously growing a number of different culture . also , in the case of the tampon , the devices incorporated therein may have , respectively , different culture mediums for indicating the presence of different micro - organisms .

for use in testing for the presence of micro - organisms , a tampon , and a plurality of devices incorporated therein , said devices each include a culture medium for promoting the growth of micro - organisms and a carrier means for carrying the culture medium .

fig1 illustrates the main components of a laser system constructed in accordance with the present invention for use in ablating tissue , shown at t . thus , the illustrated system includes a laser 2 which produces a continuous laser beam 4 . in the preferred embodiment of this invention the laser 2 is a carbon dioxide laser . the continuous laser beam is applied to a laser scanner system , generally shown by box 6 , which cyclically scans the beam along two orthogonal axes to cause the beam to trace lissajous figures , shown generally at 8 in fig1 over the tissue t to be ablated . the laser beam leaving the scanning system 6 first passes through a focussing lens 10 which focusses the laser beam on tissue t . the scanning system 6 includes two mirrors 12 , 14 , each rotated by a motor m 1 . m 2 . these mirrors are so located with respect to the laser beam 4 and also to each other to cyclically scan the laser beam along two orthogonal axes , and to cause the beam to trace the lissajous fig8 over the tissue t to be ablated . the manner in which the lissajous fig8 are produced by the scanning mirrors 12 , 14 will now be described particularly with reference to fig2 - 7 . fig2 illustrates a system including motor m1 rotating at angular velocity ω1 about an axis defined by the normalised vector b1 . a mirror ( e . g ., 12 , fig1 ) is fixed to the motor such that its normal , defined by n1 , lies at an angle of θ / 2 to the rotating axis b1 . as the motor rotates , the vector n1 defines a cone of half angle θ / 2 . the axis of symmetry of the cone is defined by the vector b1 . a ray , defined by vector a , impinging on the mirror at an angle of 45 ° to axis b1 will , according to the laws of reflection , produce reflected rays described by the time dependent vector c1 ( t ). this vector c1 ( t ) traces an envelope of a cone with an elliptical base . the vector z1 , which represents the axis of this cone , lies in the plane defined by vectors a and b1 . the angle between vectors z1 and b1 is also 45 degrees . a cartesian coordinate system based on the three vectors x1 , y1 and z1 may now be defined . the origin of this coordinate system is represented by 0 1 in fig2 . the vector x1 lies in the plane containing vectors a , b1 and z1 , and is perpendicular to vector z1 . the direction of vector y1 is perpendicular to vectors x1 and z1 . the projections of the reflected rays can now be described by the following equations : a x1 ( t ) is the angle of the projection c1 ( t ) in the plane x1 - z1 ; a y1 ( t ) is the angle of the projection of c1 ( t ) on the plane y1 - z1 ; and δ1 is an arbitrary phase which defines the angles a at time t = 0 . the relatively large displacement associated with amplitude θ in equation ( 1 ) lies in the plane containing vectors a and b1 . the smaller amplitude θ /√ 2 of equation ( 2 ) is in the direction of vector y1 . now can be added a second motor m2 ( fig3 ), whose axis is defined by vector b2 rotating with angular velocity ω2 . a mirror ( e . g ., 14 , fig1 ) whose normal is n2 ( t ) is fixed to motor m2 forming an angle of θ / 2 between normal n2 ( t ) and vector b2 ( as in motor m1 ). motor m2 will be aligned such that the axis of vector b2 lies at 45 ° to the axis of vector z1 . vector b2 also lies in the plane defined by vectors z1 and y1 . as a result , there is obtained reflected rays c2 ( t ) which form a solid cone with a circular ( not elliptical ) base . the axis of symmetry z2 of this cone lies at 45 ° to the axis b1 of the motor m1 and in the plane defined by vectors z2 and b2 . a new cartesian coordinate system may now be defined having an origin at 0 2 ( see fig3 ). vector x2 is perpendicular to vector z2 and lies in the plane defined by vectors b2 and z2 . vector y2 is perpendicular to vectors x2 and z2 . the larger amplitude always exists in the x direction and the smaller amplitude in the y direction . the two motors m1 , m2 are aligned in such a way that the x direction of motor m1 combines with the y direction of motor m2 , and the y direction of motor m 1 combines with the - x direction of the second motor . in this way amplitude compensation is obtained , resulting in a cone with a circular ( not elliptical ) base . all the rays c2 ( t ) exiting from the second mirror ( e . g ., 14 , in fig1 ) are defined by the following equations : ## equ1 ## assuming δ1 =- 90 ° and δ2 = 0 then : the angle of the exiting rays formed with axis z2 can exist between zero and ( θ + θ /√ 2 ). thus the rays fill the whole area of the base of the cone whose half angle is defined by ( θ + θ /√ 2 ). a ray which is focussed by a lens of focal length &# 34 ; f &# 34 ; ( e . g ., lens 10 , fig1 ), will be displaced at the back focal plane of the lens by an amount a . f , where a is the angle subtended by the ray and the optical axis of the lens ( see fig4 ). if a lens is placed perpendicular to axis z 2 ( fig5 ), a time dependent ray pattern will be produced at the focal plane of the lens ( of focal length f ), given by the following equations ( see fig5 and 6 ): . sub . x2 ( t )= fa . sub . x2 ( t ) = f ( θ /√ 2 ) cos ( ω . sub . 1 * t )+ fθ cos ( ω . sub . 2 * t ) eq . 7 . sub . y2 ( t )= fa . sub . y2 ( t ) = fθ sin ( ω . sub . 1 * t )+ f ( θ /√ 2 ) sin ( ω . sub . 2 * t ) eq . 8 for example , the lens may be of f = 125 mm ; the mirror wedge angle may be θ = 2 . 34 mrad ; and the angular velocities may be ω1 = 600 rad / sec and ω2 = 630 rad / sec . let a = θf /√ 2 = 0 . 207 ; b = θf = 0 . 293 ; and c = ω2 / ω1 = 1 . 05 . the ray exiting from the lens will scan at the focal plane an area whose limits are defined by a circle of radius 0 . 5 mm ( see fig6 ). every 20 revolutions the ray completely scans the whole area and starts anew . the 20 revolution scan period is about 0 . 2 seconds . the resultant ray pattern can be seen in fig6 . fig8 illustrates the invention in one form of laser apparatus used for free - hand surgery . the laser apparatus illustrated in fig8 therein designated 20 , outputs a laser beam via an articulated - arm system 22 and a handpiece 24 grasped by the surgeon for directing the laser beam to the appropriate locations of the tissue t to be ablated in accordance with the present invention , the laser of fig8 includes a scanner system , generally designate 26 , as described above for cyclically scanning the continuous laser beam along two orthogonal axes and thereby to cause the beam to trace lissajous figures over the tissue t to be ablated . in the apparatus illustrated in fig8 the focussing lens ( 10 , fig1 ) is in the hand - held handpiece gripped and manipulated by the surgeon . following is one example of the parameters of a hand laser apparatus such as illustrated in fig8 . these two velocities , vavg . and vmin ., are four times that shown in fig7 because fig7 represents a scan radius of 0 . 5 mm , whereas in the above example the radius is four times greater . 1 . spot radius at focus ## equ2 ## 2 . power density at focus p . d .= p / s = p / π · w0 2 = 637 watts / mm &# 39 ; where s is the area of the focussed spot . at this power density the thermal damage is minimal , and there are no signs of carbonisation . assuming no scanning , the rate of evaporation ve would be : ## equ3 ## at such a large speed there is no way of controlling the homegeneity of tissue removal . as a result , deep holes and valleys are formed . if the scanner is operated at a scan radius of r = 2 mm , the average power density on the tissue within the scanning area is : at this speed it is easy to control the rate of tissue removal causing minimal damage . because of the scan speed , each element of the tissue feels the equivalent of a short time pulse . the pulse duration is given by the ratio of the spot diameter at the focus ( 2wo ) to the linear scan speed ( vs ) ( see fig7 ). pulses of this duration give very low thermal damage . the time between successive pulses is 0 . 2 sec . this is the ideal time for the tissue to cool down . this is a further reason for low thermal damage . fig9 illustrates the invention included in another form of laser apparatus particularly useful for microsurgery . the laser , generally designated 30 in fig9 outputs a laser beam via a system of articulated arms 32 and a micro - manipulator 34 , such as described in u . s . pat . no . 4 , 228 , 341 , to the tissue t to be ablated . micro - manipulator 34 includes a joystick 35 enabling the surgeon to manipulate the laser beam as desired , and also an eyepiece and microscope ( not shown ) to permit the surgeon to view the working area containing the tissue to be ablated . the scanner system , generally designated 36 , corresponds to the scanner system 6 in fig1 and is effective to cyclically scan the continuous laser beam along two orthogonal axes as described above to cause the beam to trace lissajous figures over the tissue to be ablated . following is one example of the parameters of a gynecological colposcope constructed as illustrated in fig9 and having a working distance of 400 mm . the laser apparatus illustrated in fig9 and constructed in accordance with the foregoing parameters produces the following results : ## equ4 ## 2 . power density at focus p . d .= p / s = p / πwo 2 = 165 watts / mm 2 . at this power density the thermal damage and carbonisation is minimal . with the scanner , p . d .= p / πr 2 = 60 / π2 2 = 4 . 78 watts / mm 2 , and the rate of evaporation ve = 0 . 4 × p . d .= 0 . 4 × 4 . 78 = 1 . 9 mm . sec . this represents an evaporation rate which is very convenient for efficient working conditions . while the invention has been described with respect to several preferred embodiments , it will be appreciated that these are set forth merely for purposes of example , and that many variations may be made . for example , the scanning need not trace lissajous figures ; in fact , only one scanning mirror is needed since the movement of the laser by the surgeon will cause the beam to scan the surface to be ablated . also , more than two mirrors could be used . many other variations , modifications and applications of the invention may be made .

a system for causing uniform ablation of irradiated material of living tissue while not causing damage below a predetermined depth where laser radiation is provided sequentially and continuously in a predetermined pattern is disclosed .

the compositions of one embodiment of this subject invention contains effective amounts of tumor necrosis factor ( tnf ), human interferon , an antiviral surface active agent , and a physiologically acceptable carrier . the tnf may be natural or synthetic tnf , i . e . tnf produced by recombinant dna technology . the compositions may also contain any , natural or synthetic , of the known varieties or sub - varieties of human interferon . thus , the compositions may contain natural and / or synthetic tnf and natural and / or synthetic alpha - or human leukocyte interferon , and / or beta - or fibroblast interferon , and / or gamma - or immune interferon . an effective dose of tnf for treatment in accordance with the practices of the subject invention would be about 10 2 to 10 8 iu , more or less . an effective dosage of human interferon when present in the compositions in accordance with the practices of the subject invention would be about 10 2 to 10 8 , preferably about 10 4 to 10 8 , i . u . although tnf is the preferred antitumor agent in the practices of this invention , particularly in combination with human interferon , e . g . alpha - and / or beta - and / or gamma - interferon , other antitumor agents in combination with the antiviral surfactant , alone or in combination with interferon or with another antitumor agent are also useful . as indicated hereinabove , other antitumor agents usefully employed include interleukin , especially interleukin ii , tclf and cbf , separately or in any combination . these other antitumor agents are employed in an effective antitumor amount , equivalent to or the same amount as indicated hereinabove with respect to interferon and / or tnf . antiviral surfactants useful in the compositions of this invention include the antiviral anionic , cationic , and nonionic surfactants . antiviral surface active agents are known , see u . s . pat . nos . 4 , 147 , 803 , 4 , 020 , 183 , 4 , 139 , 630 and 4 , 507 , 281 . suitable anionic surfactants include sodium alkylsufonates and sodium alkylbenzenesulfonates . suitable cationic surfactants include quarternary ammonium detergents , such as cetyl pyridinium chloride , and benzalkonium chlorides . nonionic surface active agents are preferred in the compositions of this invention . in contrast to cationic , anionic , and ampholytic surface active agents , the nonionics surface active agents or surfactants contain no ionizable groups and have no surface charge . they depend upon the entire molecule for surface activity . almost any hydrophobic compound which has in its structure a carboxy , hydroxy , amido or amino group with a free hydrogen attached to the nitrogen , can be reacted with ethylene oxide to form a nonionic surfactant . at least three groups of nonionic surfactants are recognized ( a ) those having an ether linkage between the hydrophilic and hydrophobic portions of the molecule , ( b ) those having an ester or etherester linkage , and ( c ) those having an amide linkage . nonionic surfactants having at least one ether or amide linkage are preferred for purposes of the present invention . examples of preferred nonionic surfactants include the following : nonylphenoxypolyethoxy ethanol ( available under the trade name nononxynol - 9 ), p - diisobutylphenoxypolyethoxy ethanol ( available under the trade name triton x - 100 ), polyoxyethylene ( 10 ) oleyl ether ( available under the trade name brij - 97 ), and onyx - ol ( available under the trade name onyx - ol 345 ). for an effective dosage the antiviral surface active agent for the compositions and purposes of this invention comprises about 0 . 1 % to 20 % by weight of the pharmaceutical composition . the preferred range is about 1 % to 5 % by weight . the balance of the pharmaceutical compositions comprises a substantially inert , physiologically acceptable carrier . the carrier should not react with the active ingredients and not to reduce their effectiveness . suitable physiologically acceptable carriers include water , ethanol , polyethylene glycol , mineral oil , petrolatum , propylene glycol , dimethylsulfoxide , and the like . dimethylsulfoxide is also usefully included in the compositions since diemthylsulfoxide is known to effectively carry drugs past the dermal layer of the skin and as such would be a useful component of the carrier for the composition of this invention . the compositions of this invention , as indicated , are topically applied or administered in formulations suitable for topical application , such as gels , creams , lotions , shampoos , sprays and the like . the following are examples of suitable formulations of carriers useful in the preparation of compositions in accordance with this invention : ______________________________________pharmaceutical lotionpropylene glycol 24 . 75 ml . triethnolamine 1 . 00 ml . water 7 . 00 ml . oleic acid 1 . 50 gm . polyethylene glycol monostearate 10 . 50 gm . silicon fluids 10 . 00 ml . carbopol - 934 ( 2 % mucilage ) 50 . 00 ml . pharmaceutical cream awhite petrolatum 41 . 00 gm . microcrystalline wax 3 . 00 gm . fluid lanolin 10 . 00 gm . sorbitan monooleate 4 . 75 gm . polysorbate - 80 0 . 25 gm . purified water 41 . 00 gm . pharmaceutical cream bspermaceti 7 . 5 % white wax 12 . 0 % mineral oil 56 . 0 % sodium borate 0 . 5 % sorbitan monooleate 5 . 0 % water 19 . 0 % ______________________________________ in addition to the components listed above , the antitumor agent , e . g . tnf , interleukin ii , tclf and / or cbf , preferably in combination with a human interferon in an amount from about 10 2 iu to 10 8 iu , would also be included . additionally , there would be included an antiviral surfactant in the amount 0 . 05 % to about 20 % by weight based on the overall composition and , if desirable , a minor amount of dimethylsulfoxide , e . g . an amount in the range about 0 . 1 - 10 % by weight of the overall composition . topical administration of compositions of the present invention may be effected by applying a small amount ( e . g ., about 1 - 5 mls ) of the compositions directly to and onto the areas adjacent to the site of the lesion with a cotton swab , soft brush , sponge or the like . a quantity sufficient to cover the lesion is usually adequate for treatment . treatment by topical application of the composition should be regular and , if necessary , frequent , for example , every 2 - 4 hours , for about 1 - 7 days , more or less . the compositions also display antimicrobial activity as well as antitumor and antiviral activity . for example , the compositions are effective in treating certain bacterial infections . as used herein , the term antimicrobial activity refers to activity against microorganisms other than viruses , such as bacteria , yeast and fungi . in addition to direct topical application of the compositions , the composition may be administered topically by various other methods , for example , by application to the affected skin area in microencapsulated , temperature and / or pressure sensitive form or encapsulated in a film or solid carrier which is soluble in body fluids and the like for subsequent release of the compositions . the compositions may also be delivered in a foam , spray , tampon , suppository , etc . in another embodiment , the antitumor agent is incorporated in a cosmetic composition containing effective amounts of interferon , the antiviral surfactant , and the antitumor agent , such as tnf and / or interleukin ii , and a physiologically acceptable cosmetic carrier . additional components , for example , skin softeners , may be included in the cosmetic formulations . cosmetic formulations are known in the art and are usually hypoallergenic and ph controlled . the cosmetic formulations of this invention are useful as a prophylactic or for the cosmetic and therapeutic treatment of cancerous skin disorders . cosmetic formulations according to the present invention generally contain less antitumor agent , human interferon and antiviral surface active agent than the usual therapeutic preparations . the preferred range of the antitumor agent in such compositions is about 10 2 to about 10 8 iu the preferred range of human interferon is 10 3 - 10 5 i . u . and the preferred range of the antiviral surface active agent is 0 . 1 %- 5 %. a typical carrier for use in a cosmetic formulation according the the present invention has the formulations : ______________________________________cosmetic cream______________________________________beeswax 12 . 1 % spermaceti 12 . 6 % sweet almond oil 54 . 4 % borax 0 . 5rose water 19 . 4 % ______________________________________ a composition in the form of an aqueous gel and containing 2 % by weight nonylphenoxypolyethoxy ethanol and a minor amount , about 10 5 i . u ., tnf is applied topically to a cancerous skin lesion , such as a melanoma skin lesion , sufficient to generously cover the skin lesion . after 4 hours another similar application is made and this procedure continued for four days and then discontinued for observation and evaluation before another such series of treatment . a composition in accordance with example 1 is prepared but additionally comprising a minor amount , about 10 5 i . u . interleukin ii , and is applied similarly to a melanoma skin lesion . a composition in accordance with example 1 is prepared but additionally comprising a minor amount , about 10 5 i . u ., human alpha interferon , and is applied similarly to a melanoma skin lesion . as would be apparent from the above , the compositions of this invention are also useful when topically applied in the treatment of disseminated or metastasized cancer or tumors . accordingly , topical application of the composition provides another approach or technique for , in effect , the systemic treatment of cancer or tumors , rather than the conventional intravenous technique . all the above - identified u . s . patents and the above cited publication are herein incorporated and made part of this disclosure . as is apparent to those skilled in the art , many changes and modifications and substitutions are possible in the practices of this invention without departing from the spirit or scope thereof .

a composition containing an antiviral and an antitumor agent is useful when topically applied for the treatment of tumors and cancers , particularly for the treatment of viral or cancerous skin disorders and skin manifestations thereof . interferon is usefully employed in these compositions in combination with the antitumor agent . antitumor agents which are useful include interleukin , such as interleukin ii , tumor necrosis factor , target cell lysis factor and carcino - breaking factor . these compositions are especially usefully applied by topical application to the skin manifestation of the viral or cancerous skin disorder .

fig1 shows a schematically simplified blood circuit 1 with a dialysis filter or blood filter or filter 3 for executing an extracorporeal blood treatment in which blood coagulation is influenced by a citrate - calcium administration . the filter 3 is connected with an apparatus 4 ( indicated by broken lines in fig1 ) according to the present invention for extracorporeally treating blood . the blood circuit 1 encompasses an arterial patient line or line 5 that leads away from a patient p with a blood pump 7 . from a citrate solution source , here exemplarily embodied as a citrate bag 9 , a citrate solution is delivered into the line 5 . from the citrate solution source , four percent na 3 citrate , for example , is supplied . the blood circuit 1 further encompasses a venous patient line or line 11 that leads to the patient p . a dispensing device , here embodied as calcium pump 12 , is provided for delivering a calcium solution into the line 11 from a calcium solution source , in fig1 exemplarily embodied as a calcium bag 13 . from the calcium solution source , a cacl 2 solution , for example , is supplied . this solution may have a calcium concentration of 91 mmol / l . the filter 3 is connected with a dialysate source , e . g ., a dialysate bag 15 , on the dialysate side . dialysate , e . g ., a citrate - calcium ( ci - ca ) dialysate k2 , is supplied to the filter 3 from the dialysate source . the filter 3 is further connected with an outlet 17 . the line 11 is further connected with a substituate source , here in the form of a substituate bag 19 . the bag 19 contains a calcium - containing substituate which may , e . g ., be the product “ multibic ” by the company fresenius medical care deutschland gmbh . the bag 19 is merely exemplarily — as an example for a source for substituate — arranged for dispensing the substituate in postdilution , i . e ., at a location of the blood circuit 1 which in flow direction of the blood is behind the filter 3 . the blood flow set by the blood pump 7 may be set to 100 ml / min . at a pump not shown in fig1 for conveying citrate solution out of the citrate bag 9 , a flow of 180 ml / h may be set . at the dispensing device 12 , here embodied as calcium pump , for conveying calcium solution out of the calcium bag 13 and into the extracorporeal blood circuit 1 , a flow of 45 ml / h may be set . at a pump not shown in fig1 for conveying dialysate out of the dialysate bag 15 , a flow of 1800 ml / h may be set . at a pump not shown in fig1 for conveying substituate out of the substituate bag 19 , a flow of 1000 ml / h may be set . the outflow may be about 3100 ml / h . its volume depends on the doctor &# 39 ; s targeted net ultrafiltration rate . it is evident from fig1 that calcium is supplied to the patient p from the calcium bag 13 , the dialysate bag 15 and the substituate bag 19 . supplying the calcium or the calcium solution from the calcium bag 13 by the calcium pump 12 is controlled or regulated by a control device 20 of the apparatus 4 . a corresponding signal connection is indicated as dashed - dotted line . fig2 exemplarily shows possible setting ranges for a calcium dosage with the purpose of calcium balance . fig2 further shows limits of the setting range . the dimension of the dosage for calcium , in fig2 denoted as calcium dose , is millimole per liter [ mmol / l ]. the setting range regarded as permissible or the allowed setting range for the dosage of calcium for the calcium balance in the example of fig2 extends over the range of values from 0 to 3 mmol / l . determining a calcium dose in a range 21 corresponds to the substitution of an amount of calcium which is smaller than the expected amount of calcium that will be removed by the blood treatment . a setting in this area might lead to an undesired calcium loss , therefore setting such dosage may in certain embodiments according to the present invention take place together with displaying a message , an optical or acoustical alarm or the like . alternatively , trying to set a calcium dose in the range 21 may also be a sign that less calcium than expected goes out of the blood through the membrane and is removed with the effluent . this may be a result of a reduced permeability of the membrane . this possibility may be pointed out to the user or the treatment apparatus in certain embodiments according to the present invention . setting a calcium dose which is in a range 23 would match the expectations and would not lead to a message for the user . setting a calcium dose which is in a range 25 corresponds to a calcium substitution above the expected calcium removal through the effluent . a message for the user about this is in certain embodiments according to the present invention provided ; in others , such message is not output . it might also be pointed out that possibly a citrate accumulation is present , as it is often connected with an increased calcium substitution demand . setting a calcium dose which is in a range 27 would be regarded as being too high and would not be permissible . selectability of such impermissible value for a calcium balance by the device or pump according to the present invention would not be possible according to the present invention . the user may in some embodiments according to the present invention , should he try to set calcium dose in the range 27 , receive the message that he should check whether under the given circumstances a different treatment option than the set one or executed one is more suitable and optionally set this option . further , in some embodiments according to the present invention , an automatic adjustment of other parameters than the calcium dose may take place which may lead to a balanced calcium balance . a list with automatically permissible changes may be on file . if adjustments are necessary or even automatically performed , it is in certain embodiments provided to inform the operator about this . fig3 shows a user interface 31 according to the present invention for executing a method according to the present invention . from the left side of the user interface 31 indications regarding the arterial pressure 33 , the venous pressure 35 and the transmembrane pressure 37 , here given each in mmhg , may be taken . the corresponding values may hereby be read both expressed as numbers and as variable markings on scales 39 . the user interface 31 further comprises an indication 41 of the currently set or measured blood flow , an indication 43 of the ultrafiltration rate , an indication 45 if a continuous heparin administration takes place ( here with 0 for off ), and an indication 47 if a heparin administration takes place via bolus ( here with b for “ bolus ” set ). the user interface 31 further comprises setting possibilities 49 for a level of the venous bubble trap and control interfaces 51 for displaying or controlling further treatment data or treatment options . additionally , the user interface 31 comprises a display 53 for displaying the set dialysate flow ( corresponding to the dialysate rate ), a display 55 for displaying the set citrate flow in relation to the blood flow , and a display 57 for displaying the set calcium flow in relation to the filtrate flow . all or some of the displays mentioned above or hereafter may in certain embodiments according to the present invention also serve for changing the set flow rates . for example , they may be adjustable by touching the corresponding display surfaces . in the above description , the user interface 31 does not differ from an already known user interface as may be used also for a different treatment option such as the cvvhd ( continuous veno - venous hemodialysis ) in combination with citrate - calcium anticoagulation . what is new or added , however , is the display 59 of the substituate flow . the user interface 31 thus additionally encompasses the indication which is relevant for executing a method known as cvvhdf ( continuous veno - venous hemodiafiltration ) in combination with citrate - calcium anticoagulation , as is indicated in the upper left corner of the user interface 31 . the user interface 31 illustrated in fig3 is thus a development of a user interface as may be used with the cvvhd . the user interface 31 according to the present invention therefore advantageously differs only marginally from a user interface the user already knows from another blood treatment option . it may therefore in certain embodiments according to the present invention be easily adjusted based on a display for cvvhd to a display for cvvhdf , and vice versa . offering a few specific displays and / or setting possibilities , more or less depending on which of the before - mentioned treatment options should be followed , may suffice . coupled with this is the advantage that the user may use a user interface that is already known to him from performing a treatment option in an only marginally changed setup of the user interface also when performing a further treatment option or the method according to the present invention . confusing the user may thus be prevented and the requirement that the user has to be familiar with a multitude of user interfaces can be eliminated . if an impossibility of a set constellation of the various setting parameters ( flows , calcium dose , and so on ), e . g ., in the sense of an in principle negative value of the calculated calcium flow out of the calcium pump , is produced when the requested value for the calcium addition by the calcium pump is set by the user , the calcium dose is in certain embodiments according to the present invention increased to zero , to the minimum settable value , a lower threshold value or to a predetermined default value that deviates herefrom . the minimum settable value is in some embodiments according to the present invention a value rounded up to a full 0 . 1 mmol / l . it may , e . g ., be calculated according to the formula ( 1 ) or ( 1 ′) given above or in a different suitable way . such technical impossibility of the required calcium dose ( i . e ., for instance , a negative value ) that was determined by way of calculation may , e . g ., also be taken into account in that the calcium dose that is required but not realizable in practice cannot be entered by the user . for example , the display computer , or every other suitable device , may already be programmed accordingly in order to make only useful and / or technically possible dosages selectable to the user . if the user nevertheless enters technically impossible values or at least values that are not allowed for the calcium dosage by the calcium pump , which may initially be possible in certain embodiments according to the present invention , however , not in others from the beginning , preferably a message may be output in order to inform the user of , e . g ., the impossibility . in certain embodiments according to the present invention , a message for the user is output whenever a setting of the calcium dose that was set on the machine side , i . e ., automatically , deviates from the one the user has requested or has tried to enter . this may contribute to the user &# 39 ; s understanding of the machine &# 39 ; s behavior . in some embodiments according to the present invention , the calcium dose may automatically be set to zero , a minimum value or the value resulting by way of calculation ( e . g ., according to formula ( 1 ) or ( 1 ′)), if or when setting parameters other than the calcium dose , such as for example , the dialysate flow or the substituate flow , which influence the calculated calcium flow , change — with or without the involvement of the user . the user may be informed about this by a message , alarm , text field , and so on . an exception from the latest mentioned procedure is in some embodiments according to the present invention provided when or if after having achieved the balance aim , i . e ., an aim for the cumulated removal of water from the blood , a stop or an end of the ultrafiltration is reached and therefore the machine transitions to a changed state ( in relation to parameter value , flows and the like ). in this case , the calcium dose does not have to be adjusted in certain embodiments according to the present invention . also in this situation , the user may be informed hereof by a message , alarm , text field , and so on . in certain embodiments according to the present invention , it is provided that before an automatic change of parameter values , e . g ., for the calcium pump , valid settings for arbitrary setting parameters of the device or apparatus according to the present invention ( calcium dose and others ) are automatically saved by the device or apparatus according to the present invention . using the saved values , the machine &# 39 ; s parameters may be reset — automatically or at the user &# 39 ; s suggestion — to the previous settings when the previous settings are again technically and medically useful , possible and desired . the user may receive a message that due to certain changes previous values may now be set again . examples for such settings and parameters include the calcium dose , the blood flow , the dialysate flow , the substituate flow and others , without being limited hereto . in some embodiments according to the present invention , when starting the treatment , a flag set in a previous treatment , which , e . g ., is always set when the calcium dose was increased automatically , may first be initialized as deleted . the flag may be set when the user enters a calcium dose . by setting the flag , the previously valid value of the calcium dose is at least temporarily saved . if or when the flag is set and if or when decreasing the calcium dose is possible or increasing the calcium dose is necessary or advisable after changing another parameter ( optionally except for automatically decreasing the ultrafiltration rate to zero when the balance aim is achieved ), a message to the user may be output that he may now decrease the calcium dose . the flag may subsequently be deleted . alternatively , such adjustment may also take place automatically . in some embodiments according to the present invention , the selection of the calcium dose is limited to a countable finite number of dose levels . in these embodiments , setting or controlling the lowest possible calcium dose , as it was calculated , e . g ., by formula ( 1 ) or ( 1 ′), is not provided or not provided in each case . in these embodiments , e . g ., 0 . 1 mmol / l or 0 . 2 mmol / l may be provided as setting width or setting increments which has advantageously proven effective . therefore , in some embodiments according to the present invention , an increment of 0 . 1 mmol / l or 0 . 2 mmol / l is provided for the available setting steps . the limitation of the possible adjustment by or to at least 0 . 1 mmol / l or 0 . 2 mmol / l advantageously allows for a little time - consuming preparation of the hardware utilized in the treatment , in particular of the dispensing device by which the calcium balance takes place , and its control . furthermore , this allows for a design of the user interface in which the smallest setting steps on the one hand enable individualization of the treatment , and on the other hand prevent displaying unnecessary information by waiving precision of the display that goes beyond this , which would be necessary with smaller setting steps . this advantageously makes the user interface clearer . hereafter , three examples of the method according to the present invention are illustrated . in an exemplary initial configuration of the apparatus according to the present invention , the following applies : hereby , the internally calculated flows for the citrate solution is 176 . 47 ml / h and for the calcium solution is 37 . 95 ml / h . in case the operator reduces the calcium dose now to , e . g ., 1 . 2 mmol / l , the calcium flow changes to 22 . 18 ml / h with otherwise unaltered values . informing the operator with the text “ low calcium dose ” may be advisable — the filter may have lost function . if the calcium dose is reduced to 0 . 5 mmol / l , the calcium flow decreases to 0 . 38 ml / h with otherwise unaltered values . informing the operator with the text “ lower limit for calcium dose reached ” may be advisable . if the calcium dose is reduced to 0 . 4 mmol / l , the calcium flow would by way of calculation have to be decreased to − 2 . 70 ml / h to achieve a correct calcium balance with otherwise unaltered values . however , such a setting is obviously useless and is therefore not offered or permitted . in a different exemplary initial configuration of the apparatus according to the present invention , the following applies : hereby , the internally calculated flows for the citrate solution is 176 . 47 ml / h and for the calcium solution is 6 . 58 ml / h . the calcium dose is hereby low ; however , it is still permissible due to the high dialysate flow . yet , by reducing the dialysate flow , a limit of the calcium dose may be reached , as can be seen hereafter . if the operator reduces the dialysate flow to , e . g ., 800 ml / h , the calculated calcium flow changes to − 0 . 47 ml / h with otherwise unaltered values . automatically raising the calcium dose to a lowest of the possible values is advisable here , informing the operator if applicable . after the calcium dose was automatically raised to , e . g ., 0 . 8 mmol / l , the calcium flow increases to 1 . 62 ml / h . this is a permissible value . in a further different exemplary initial configuration of the apparatus according to the present invention , the following applies : hereby , the internally calculated flows for the citrate solution is 176 . 47 ml / h and for the calcium solution is 1 . 71 ml / h . the calcium dose is hereby low ; however , it is still permissible due to the low substituate flow . yet , by raising the substituate flow , a limit of the calcium dose may be reached , as can be seen hereafter . if the operator raises the substituate flow to , e . g ., 1000 ml / h , the calcium flow calculated to be required for achieving a calcium balance changes to − 2 . 70 ml / h with otherwise unaltered values . automatically raising the calcium dose to a lowest of the possible values is here also advisable , informing the operator if applicable . after the calcium dose was automatically raised to , e . g ., 0 . 5 mmol / l , the calcium flow increases to 0 . 38 ml / h with otherwise unaltered values . this is a permissible value .

the present invention relates to a method for controlling or regulating an apparatus for extracorporeally treating blood in an extracorporeal blood circuit while adding citrate and calcium for the purpose of anticoagulation . the apparatus comprises a dispensing device for dispensing a calcium solution into the extracorporeal blood circuit . the method encompasses outputting a signal to the dispensing device for altering a setting of the dispensing device , in which the setting corresponds to or effects an addition of calcium , a calcium dosage , concentration , amount or rate . the method further encompasses defining or altering the signal with regard to a calcium content or a calcium concentration of a substituate solution used in extracorporeally treating the blood , and detecting the range of values of permissible or allowed settings for the calcium introduced or to be introduced into the extracorporeal blood circuit by the dispensing device , particularly its amount , concentration or dosage .

surprisingly , it has now been found that the compounds described below in greater detail have a high affinity for sigma receptors and are therefore advantageously suitable for the treatment of illnesses which can be favorably affected by alterations in the function of sigma receptors . the invention relates to the use of compounds of the formula i ## str2 ## in which either r4 is cyclopentyl , cyclohexyl , phenyl , 2 - norbornen - 5 - yl or tricyclo [ 2 . 2 . 1 . 0 2 , 6 ] hept - 2 - yl , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . 1 - 4c - alkyl represents straight - chain or branched alkyl radicals having 1 to 4 carbon atoms . examples which may be mentioned are the butyl , isobutyl , sec - butyl , tert - butyl , propyl , isopropyl , ethyl and , in particular , the methyl radical . suitable salts of compounds of the formula i are preferably all pharmacologically tolerable acid addition salts with the inorganic and organic acids customarily used in pharmacy . those which are suitable are water - soluble and water - insoluble acid addition salts with acids such as , for example , hydrochloric acid , hydrobromic acid , phosphoric acid , nitric acid , sulfuric acid , acetic acid , citric acid , d - gluconic acid , benzoic acid , 2 -( 4 - hydroxybenzoyl ) benzoic acid , butyric acid , sulfosalicylic acid , maleic acid , lauric acid , malic acid , fumaric acid , succinic acid , oxalic acid , tartaric acid , embonic acid , stearic acid , toluenesulfonic acid , methanesulfonic acid or 3 - hydroxy - 2 - naphthoic acid , it being possible to employ the acids in salt preparation - depending on whether it is a mono - or polybasic acid and depending on which salt is desired -- in an equimolar quantitative ratio or one differing therefrom . depending on the nature of the substituents , the compounds of the formula i can be optically active compounds . the invention therefore includes both the enantiomers and their mixtures and racemates . illnesses which may be mentioned which are favorably affected by substances having affinity for sigma receptors are psychoses ( schizophrenia ), hallucinations in combination with psychotic disorders and chronic psychological depressions , psychoneuroses , brain function disorders ( cerebral ischemia , cognitive dysfunction ), disorders of intestinal function ( absorption , secretion , motility ) and of other smooth muscular organs , such as vas deferens and bladder ( urinary incontinence ), and other disorders such as are connected with affinities for sigma receptors and such as are described , for example , in the literature surveys of b . l . largent et al . ( eur . j . pharmacol ., 155 , 345 - 7 , 1988 ), s . i . deutsch et al . ( clinical neuropharmacology , 11 ( 2 ), 105 - 119 , 1988 ), t . p . su ( eur . j . biochem ., 200 , 633 - 642 , 1991 ) and j . m . walker et al . ( pharmacol . rev . 42 , 355 - 402 , 1990 ). in the use according to the invention of the compounds of the formula i for the production of the abovementioned medicaments , the pharmacologically active compounds of the formula i and / or their salts (= active compounds ) are processed with suitable pharmaceutical auxiliaries or excipients to give tablets , coated tablets , capsules , suppositories , patches ( e . g . as tts ), emulsions , suspensions or solutions , the active compound content advantageously being between 0 . 1 and 95 % and it being possible to achieve a pharmaceutical administration form ( e . g . a delayed release form or an enteric form ) exactly matched to the active compound and / or to the desired onset of action by the appropriate choice of the auxiliaries and excipients . the person skilled in the art is familiar on the basis of his expert knowledge with the auxiliaries or excipients which are suitable for the desired pharmaceutical formulations . apart from solvents , gel - forming agents , suppository bases , tablet auxiliaries and other excipients , it is possible to use , for example , antioxidants , dispersants , emulsifiers , antifoams , flavor corrigents , preservatives , solubilizers , colorants or in particular permeation promoters and complexing agents ( e . g . cyclodextrins ). the active compounds can be administered orally , rectally , parenterally or percutaneously . in general , it has proven advantageous in human medicine to administer the active compound ( s ) in the case of oral administration in a daily dose from approximately 0 . 01 to approximately 20 , preferably 0 . 05 to 5 , in particular 0 . 2 to 2 . 0 , mg / kg of body weight , if appropriate in the form of several , preferably 1 to 4 , individual doses to achieve the desired result . in a parenteral treatment , it is possible to use similar or ( in particular in the intravenous administration of the active compounds ) generally lower doses . any person skilled in the art can easily determine the optimum dose necessary in each case and manner of administration of the active compounds on the basis of his expert knowledge . if the compounds of the formula i and / or salts are employed for the treatment of sigma receptor - modulated illnesses , the pharmaceutical preparations can also contain one or more pharmacologically active constituents of other pharmaceutical groups . the invention furthermore includes the use of the compounds according to the invention for the treatment of sigma receptor - modulated illnesses . one embodiment of the invention is the use of compounds of the formula i in which a further embodiment of the invention is the use of compounds of the formula i in which r4 is cyclopentyl , cyclohexyl , phenyl , 2 - norbornen - 5 - yl or tricyclo [ 2 . 2 . 1 . 0 2 , 6 ] hept - 2 - yl , a further embodiment of the invention is the use of compounds of the formula i in which r1 is phenyl , r2 is phenyl , r3 is hydrogen , r4 is methyl , r5 is methyl , m is the number 2 and n is the number 0 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is phenyl , r2 is phenyl , r3 is methyl , r4 is methyl , r5 is methyl , m is the number 2 and n is the number 0 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is 2 - norbornen - 5 - yl , r5 is hydroxyl , m is the number 2 and n is the number 2 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is cyclopentyl , r5 is hydroxyl , m is the number 2 and n is the number 2 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is phenyl , r5 is hydroxyl , m is the number 2 and n is the number 2 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is cyclohexyl , r5 is hydroxyl , m is the number 1 and n is the number 2 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is cyclohexyl , r5 is hydroxyl , m is the number 2 and n is the number 2 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is tricyclo [ 2 . 2 . 1 . 0 2 , 6 ] hept - 2 - yl , r5 is hydroxyl , m is the number 2 and n is the number 2 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . a further embodiment of the invention is the use of compounds of the formula i in which r1 is hydrogen , r2 is hydrogen , r3 is phenyl , r4 is phenyl , r5 is hydroxyl , m is the number 2 and n is the number 3 , and their salts for the production of medicaments for the treatment of sigma receptor - modulated illnesses . the compounds of the formula i are known , for example from the patent specifications mentioned in the prior art . the affinity and selectivity of the compounds described above in greater detail for sigma receptors in comparison with nmda receptors were determined following published methods ( kornhuber et al . : eur . j . pharmacol . 162 , 483 - 490 , 1989 and neurosci . lett . 163 , 129 - 131 , 1993 ). the table shows the exemplary comparison of the compound budipine with aminoadamantane derivatives , which are also employed for the therapy of parkinsonism . a higher affinity and selectivity ( lower k i value ) of budipine for sigma receptors has now surprisingly been found . ______________________________________ sigma receptors ( pentazocine nmda receptors k . sub . i values ( μm ) binding ) ( mk 801 binding ) ______________________________________budipine 2 . 0 11 . 7 amantadine 20 . 3 10 . 5 memantine 20 . 0 0 . 5______________________________________

the invention concerns the use of compounds of formula , in which r 1 , r 2 , r 3 , r 4 , r 5 , m and n have the meanings indicated herein for use in the treatment of sigma - receptor modulated disorders . ## str1 ##

the term “ oral composition ” as used herein means a product that in its ordinary course of usage is retained in the oral cavity for a time sufficient to contact substantially all of the dental surfaces and / or oral tissues for purposes of oral activity . the oral composition of the present invention may be in various forms including toothpaste , dentifrice , rinses , gels , edible films , lozenges , sprays , tooth powders , subgingival gels , or denture products . the oral composition may also be incorporated onto strips or films for direct application or attachment to oral surfaces . the term “ dentifrice ” as used herein means paste , gel , powder , tablets , or liquid formulations , unless otherwise specified , that are used to clean the surfaces of the oral cavity . the composition of the present invention is an oral composition that comprises : suitable salivating agents include those agents capable of long - lasting moisturisation in the oral cavity . it may act via nerve stimulation ( via secondary processes transduced by interaction of trigeminal and parasympathetic nerves ) and / or via osmotic pressure . suitable salivating agents include , without limitation , trans - pellitorin , spilanthol , sanshool , hydroxy α - sanshool , hydroxy β - sanshool , hydroxy γ - sanshool , sanshool i , sanshool ii , sanshoamide , piperine , chavicine , 4 -( 1 - menthoxymethyl )- 2 - phenyl - 1 , 3 - dioxolane , effervescing agents and analogs thereof . other tingling sensates can be found in u . s . pat . no . 5 , 545 , 424 which is herein incorporated by reference . the total amount of salivating agent in the composition ranges from 0 . 001 - 4 . 00 % w / w . suitable gum sensates are those compounds that are able to modulate nociceptors perception of pain or inflammatory pain sensation . some embodiments may comprise a trpv1 activator , a transient receptor potential vanilloid receptor 1 activator . these activators may not only off - set bad tastes , but may also reduce dryness perception , by limiting the mouth &# 39 ; s ability to perceive dryness . in one embodiment , the gum sensate is chosen from the group comprising vanillyl butyl ether , zingerone , capsaicin , capsiate , shoagol , gingerol , piperine , bisabolol , 4 - t - butyl cyclohexanol , such as symsitive ® or a combination thereof . the total amount of gum sensates in the composition ranges from 0 . 001 - 4 . 00 % w / w . the term “ orally acceptable carrier ” as used herein means a suitable vehicle , which can be used to apply the present compositions to the oral cavity in a safe and effective manner . such vehicle may include materials such as fluoride ion sources , antibacterial agents , anticalculus agents , buffers , other abrasive materials , peroxide sources , alkali metal bicarbonate salts , thickening materials , humectants , water , surfactants , titanium dioxide , flavor system , sweetening agents , cooling agents , xylitol , coloring agents , and mixtures thereof . the following is a non - limiting list of actives that may be used in the present invention . the total amount mentioned here should only be considered in cases where said component is indeed present in the formulation . the present invention may comprise a safe and effective amount of a fluoride compound ( e . g . water soluble ). representative fluoride ion sources include : stannous fluoride , sodium fluoride , potassium fluoride , amine fluoride , sodium monofluorophosphate , zinc fluoride , and many others . in one embodiment the composition comprises stannous fluoride or sodium fluoride , as well as mixtures thereof . fluoride ion may also be considered an antibacterial active , as it has been shown to effectively reduce bacterial activity in the mouth . the total amount of fluoride compound in the composition ranges from 0 . 001 - 4 . 00 % w / w . the oral compositions may contain tartar control agents including salts of any of these agents , for example their alkali metal and ammonium salts : phosphates and polyphosphates ( for example pyrophosphates ), polyaminopropanesulfonic acid ( amps ), polyolefin sulfonates , polyolefin phosphates , diphosphonates such as azacycloalkane - 2 , 2 - diphosphonates ( e . g ., azacycloheptane - 2 , 2 - diphosphonic acid ), n - methyl azacyclopentane - 2 , 3 - diphosphonic acid , ethane - 1 - hydroxy - 1 , 1 - diphosphonic acid ( ehdp ) and ethane - 1 - amino - 1 , 1 - diphosphonate and phosphonoalkane carboxylic acids . useful inorganic phosphate and polyphosphate salts include monobasic , dibasic and tribasic sodium phosphates , sodium tripolyphosphate , tetrapolyphosphate , mono -, di -, tri - and tetrasodium pyrophosphates , sodium trimetaphosphate , sodium hexametaphosphate and mixtures thereof . the oral compositions may also contain an anticalculus agent , preferably a pyrophosphate ion source which is from a pyrophosphate salt . the pyrophosphate salts useful in the present compositions include the dialkali metal pyrophosphate salts , tetraalkali metal pyrophosphate salts , and mixtures thereof . the pyrophosphate salt may be present in one of three ways : predominately dissolved , predominately undissolved , or a mixture of dissolved and undissolved pyrophosphate . the preferred composition involves dissolved pyrophosphate with a minimum concentration of 1 % free pyrophosphate ions for clear mouthwash and dentifrice applications . disodium dihydrogen pyrophosphate , tetrasodium pyrophosphate , and tetrapotassium pyrophosphate are the preferred species . the total amount of pyrophosphate compounds in the composition ranges from 0 . 1 - 6 . 00 % w / w . free pyrophosphate ions may be present in a variety of protonated states depending on a the ph of the composition . the oral compositions may contain a buffering agent . buffering agents , as used herein , refer to agents that can be used to adjust the ph of the oral compositions to a range of about ph 3 . 0 to about ph 10 . 0 . the buffering agents include alkali metal hydroxides , ammonium hydroxide , organic ammonium compounds , carbonates , sesquicarbonates , borates , silicates , phosphates , imidazole , and mixtures thereof . specific buffering agents include monosodium phosphate , trisodium phosphate , sodium benzoate , benzoic acid , sodium hydroxide , potassium hydroxide , alkali metal carbonate salts , sodium carbonate , imidazole , pyrophosphate salts , citric acid , and sodium citrate . the total amount of buffering compound in the composition should be sufficient to attain the desired ph . preferably , it ranges from 0 . 01 - 3 . 00 % w / w . coloring agents may also be added to the present composition . pigments , pealing agents , filler powders , talc , mica , magnesium carbonate , calcium carbonate , bismuth oxychloride , zinc oxide , and other materials capable of creating a visual change to the oral compositions may also be used . titanium dioxide may also be added to the present composition . titanium dioxide is a white powder which adds opacity to the compositions . the total amount of colorant in the composition ranges from 0 . 00001 - 1 . 00 % w / w . an abrasive polishing material may also be included in the oral compositions . the abrasive polishing material contemplated for use in the compositions of the present invention can be any material that does not excessively abrade dentin . typical abrasive polishing materials include silicas , including gels and precipitates ; aluminas ; phosphates including orthophosphates , polymetaphosphates , and pyrophosphates ; and mixtures thereof . specific examples include hydrated silica , dicalcium orthophosphate dihydrate , calcium pyrophosphate , tricalcium phosphate , calcium polymetaphosphate , insoluble sodium polymetaphosphate , hydrated alumina , beta calcium pyrophosphate , calcium carbonate , and resinous abrasive materials such as particulate condensation products of urea and formaldehyde . mixtures of abrasives may also be used . the most preferred abrasive is hydrated silica . silica dental abrasives of various types are preferred because of their unique benefits of exceptional dental cleaning and polishing performance without unduly abrading tooth enamel or dentine . the silica abrasive polishing materials herein , as well as other abrasives , generally have an average particle size ranging between about 0 . 1 to about 30 microns , and preferably from about 5 to about 15 microns . the abrasive can be precipitated silica or silica gels . dental abrasives , like the dental polishing agents , also cause a small amount of enamel erosion which is termed “ polishing ” action . the removal of plaque and calculus prevents caries and periodontal disease . the polishing of teeth removes stains from tooth surfaces , but has not been shown to improve dental health over and above the effects of the removal of plaque and calculus . the total amount of abrasive polishing material in the composition ranges from 1 . 0 - 25 . 0 % w / w . additional mucoadhesive agents , such as polymeric thickeners , may be utilized . suitable mucoadhesive agents are carboxyvinyl polymers , carrageenan , hydroxyethyl cellulose , laponite and water soluble salts of cellulose ethers such as sodium carboxymethylcellulose and sodium carboxymethyl hydroxyethyl cellulose . natural gums such as gum karaya , xanthan gum , gum arabic , and gum tragacanth can also be used . mucoadhesive agents can include polymeric polyether compounds , e . g ., polyethylene or polypropylene oxide ( m . w . 300 to 7 , 000 , 000 ). a suitable class of thickening or gelling agents includes a class of homopolymers of acrylic acid crosslinked with an alkyl ether of pentaerythritol or an alkyl ether of sucrose , or carbomers . carbomers are commercially available from b . f . goodrich as the carbopol ® series . particularly the carbopols include carbopol 934 , 940 , 941 , 956 , 971p , 974p and mixtures thereof . the total amount of mucoadhesive agent in the composition ranges from 0 . 001 - 16 . 00 % w / w . a humectant can help to keep the oral composition from hardening upon exposure to air and provide a moist feel in the mouth . a humectant or additional solvent may be added to the oral composition . suitable humectants for the present invention include water , edible polyhydric alcohols such as glycerin , sorbitol , xylitol , butylene glycol , polyethylene glycol , propylene glycol , erythritol , maltitol , isomalt , lactitol , diglycerin , hydrogenated starch hydrolysate ( hsh ), and combinations thereof . sorbitol , glycerin , water , and combinations thereof are preferred humectants . the total amount of humectants in the composition ranges from 10 - 90 % w / w . suitable water retention agents are those capable of avoiding the loss of water molecules . examples of suitable agents include , without limitations , beta glucan soluble fibers , such as oat beta glucan , trimethyglycine , l - carnosine , and other aminoacids . the total amount of water retention agents in the composition ranges from 0 . 001 - 2 . 00 % w / w . a surfactant may be added to the oral composition . surfactants , also commonly referred to as sudsing agents , may aid in the cleaning or foaming of the oral composition . suitable surfactants are those which are reasonably stable and foam throughout a wide ph range . the surfactant may be anionic , nonionic , amphoteric , zwitterionic , cationic , or mixtures thereof . examples of anionic surfactants useful herein include the water - soluble salts of alkyl sulfates having from 8 to 20 carbon atoms in the alkyl radical ( e . g ., sodium alkyl sulfate ) and the water - soluble salts of sulfonated monoglycerides of fatty acids having from 8 to 20 carbon atoms . sodium lauryl sulfate ( sls ) and sodium coconut monoglyceride sulfonates are examples of anionic surfactants of this type . examples of other suitable anionic surfactants are sarcosinates , such as sodium lauroyl sarcosinate , taurates , sodium lauryl sulfoacetate , sodium lauroyl isethionate , sodium laureth carboxylate , and sodium dodecyl benzenesulfonate . mixtures of anionic surfactants can also be employed . another suitable surfactant is one selected from the group consisting of sarcosinate surfactants , isethionate surfactants and taurate surfactants . preferred for use herein are alkali metal or ammonium salts of these surfactants , such as the sodium and potassium salts of the following : lauroyl sarcosinate , myristoyl sarcosinate , palmitoyl sarcosinate , stearoyl sarcosinate and oleoyl sarcosinate . cationic surfactants useful in the present invention include derivatives of aliphatic quaternary ammonium compounds having one long alkyl chain containing from about 8 to 18 carbon atoms such as lauryl trimethylammonium chloride ; cetyl pyridinium chloride ; cetyl trimethylammonium bromide ; di - isobutylphenoxyethyl - dimethylbenzylammonium chloride ; coconut alkyltrimethylammonium nitrite ; cetyl pyridinium fluoride ; etc . nonionic surfactants that can be used in the compositions of the present invention include compounds produced by the condensation of alkylene oxide groups ( hydrophilic in nature ) with an organic hydrophobic compound which may be aliphatic or alkylaromatic in nature . examples of suitable nonionic surfactants include the pluronics ( for example , poloxamers of the form peo - ppo - peo ), polyethylene oxide condensates of alkyl phenols , products derived from the condensation of ethylene oxide with the reaction product of propylene oxide and ethylene diamine , ethylene oxide condensates of aliphatic alcohols , long chain tertiary amine oxides , long chain tertiary phosphine oxides , long chain dialkyl sulfoxides and mixtures of such materials . zwitterionic synthetic surfactants useful in the present invention include derivatives of aliphatic quaternary ammonium , phosphonium , and sulfonium compounds , in which the aliphatic radicals can be straight chain or branched , and wherein one of the aliphatic substituents contains from about 8 to 18 carbon atoms and one contains an anionic water - solubilizing group , e . g ., carboxy , sulfonate , sulfate , phosphate or phosphonate . typical alkyl dimethyl betaines include decyl betaine or 2 -( n - decyl - n , n - dimethylammonio ) acetate , coco betaine or 2 -( n - coc - n , n - dimethyl ammonio ) acetate , myristyl betaine , palmityl betaine , lauryl betaine , cetyl betaine , cetyl betaine , stearyl betaine , etc . the amidobetaines are exemplified by cocoamidoethyl betaine , cocoamidopropyl betaine , lauramidopropyl betaine and the like . the betaines of choice are preferably the cocoamidopropyl betaine and , more preferably , the lauramidopropyl betaine . the total amount of surfactant in the composition ranges from 2 . 00 %- 8 . 00 % w / w . microbeads may be used in the present formulation for providing sensorial cues . it may contain encapsulated saliva flow stimulants , encapsulated freshening agents and encapsulated cooling agents . apart from the ingredients mentioned above , the composition according to the invention may comprise conventional ingredients , such as sweeteners , flavors and soothing botanicals . the table below illustrates several toothpaste compositions according to the invention . like letters following means indicate no significant difference ( nsd ) between those means . if the letters are different , there is a significant difference between those mean scores . the results show that for several criteria analyzed , the formulations according to the invention performed better than the prior art &# 39 ; s . the results , shown in table 4 , clearly show that the difference of moisturization and salivation imparted by the formulations under analysis was statistically significant when compared with the prior art formulations .

the present invention discloses an oral composition comprising a combination of salivating agents and gum sensates in an orally acceptable carrier useful in treating and / or alleviating the symptoms associated with xerostomia . additional components common in oral compositions may also be used , in order to potentiate the effect . such components may be chosen among humectants , mucoadhesive polymers , water retention agents , antioxidants , antimicrobials , breath fresheners .

for the purpose of promoting an understanding of the present invention , references are made in the text to exemplary embodiments of a ratcheting device , only some of which are described herein . it should be understood that no limitations on the scope of the invention are intended by describing these exemplary embodiments . one of ordinary skill in the art will readily appreciate that alternate but functionally equivalent parts or components may be used . the inclusion of additional elements may be deemed readily apparent and obvious to one of ordinary skill in the art . specific elements disclosed herein are not to be interpreted as limiting , but rather as a basis for the claims and as a representative basis for teaching one of ordinary skill in the art to employ the present invention . it should be understood that the drawings are not necessarily to scale ; instead emphasis has been placed upon illustrating the principles of the invention . in addition , in the embodiments depicted herein , like reference numerals in the various drawings refer to identical or near identical structural elements . moreover , the terms “ substantially ” or “ approximately ” as used herein may be applied to modify any quantitative representation that could permissibly vary without resulting in a change in the basic function to which it is related . referring to fig1 , a recirculating ratcheting assembly is designated by the reference number 10 . the device is referred to as “ recirculating ” because it mechanically resets from the highest ratchet position when extended past the maximum height . ratcheting assembly 10 includes at least one rack 12 which is moved vertically during its operation , at least one indexing rod 37 and at least one side plate 23 . as shown in fig9 , the rack 12 as a rectangular shaped member having an upward face 18 and a second downward face 19 . the two faces are connected by a contoured sidewall 17 . as shown in fig1 , the side plate 23 has an upward face 28 and a second , downward face 29 . the two faces are connected by a contoured sidewall 27 . side plate 23 also includes a contoured slot 26 that supports cylindrical face 38 of rod 37 . fig1 shows rod 37 with cylindrical face 38 . referring to fig1 and 2 , when rod 37 is positioned in slot 26 , the cylindrical face 38 of rod 37 communicates the sidewall 27 of the side plate 23 . face 19 of rack 12 communicates with face 28 of side plate 23 . fig2 shows the rack 12 and rod 37 in the seated position . cylindrical face 38 of rod 37 rests on surface 31 of side plate 23 . the semi - circular surface 16 of rack 12 rests on the cylindrical face 38 of rod 37 . in this position , rack 12 is prevented from moving downward with respect to side plate 23 . as shown in fig3 , when rack 12 is lifted vertically , angled face 15 contacts cylindrical face 38 of rod 37 , thereby lifting rod 37 clear of the ratchet rack 12 and allowing each rack tooth to pass as rack 12 is lifted . as shown in fig2 , lowering rack 12 allows rod 37 to pass between the teeth of rack 12 and settle back onto surface 31 of side plate 23 . with rod 37 in this position , it stops rack 12 from traveling down any further thus supporting rack 12 in this location . fig4 , 5 , 6 , 7 and 8 detail resetting the ratchet assembly 10 so that it may be set to its lowest position as shown in fig8 . fig4 shows rack 12 being lifted to a point where lower surface 13 of rack 12 contacts cylindrical face 38 of rod 37 . continuing to lift rack 12 allows lower surface 13 of rack 12 to contact cylindrical face 38 of rod 37 , lifting it into the reset position as shown in fig5 . fig6 shows rack 12 being lowered with rod 37 in the reset position sitting on surface 30 of side plate 23 . as rack 12 continues downward , upper surface 14 of rack 12 will contact cylindrical face 38 of rod 37 removing it from surface 30 of side plate 23 and allowing rod 37 to drop down to surface 31 of side plate 23 . this places rack 12 in its lowest position as shown in fig8 . from this point rack 12 can be lifted vertically and rod 37 can be placed in any position on rack 12 . this process can continue until rod 37 is once again places in the reset position allowing rack 12 to be lowered into its lowest position . as shown in fig1 , the ratcheting assembly 10 is combined with a plyometrics exercise assembly 40 to provide box assembly 40 with a recirculating , ratcheting movement . exercise assembly 40 comprises at least two sub - assemblies , base assembly 40 a and box assembly 40 b . base assembly 40 a comprises at least two base side walls 41 alternately interconnected with at least two side plates 23 , at least one base plate 43 , and at least one rod 37 . box assembly 40 b comprises at least two box indicia walls 45 alternately interconnected with at least two box side walls 46 ( only one visible in fig1 ), at least one ratchet 12 , and at least one exercise platform 47 . each indicia wall 45 comprises at least one measurement indicia 48 utilized to determine how far box assembly 40 b has been raised . at least one of base side walls 41 may also comprise at least one indicia slot or groove 42 , which clearly indicates the measurement . fig1 shows separate fully assembled embodiments of base assembly 40 a and box assembly 40 b . fig1 illustrates an exploded isometric view of base assembly 40 a . each lateral side of side plate 23 comprises at least one tab 23 a . during assembly , tab 23 a is inserted into at least one slot 41 a of a base side wall 41 . side plate 23 also comprises at least one slot 23 b . during assembly , slot 23 b receives at least one tab 43 a of base plate 43 . the components of base assembly 40 a are thereby strongly interconnected . fig1 illustrates an exploded isometric view of box assembly 40 b . each lateral side of each indicia 45 and side 46 wall comprises at least one tab - and - groove configuration 45 a and 46 a , respectively . during assembly , tabs of configuration 45 a are inserted into complementary grooves of configuration 46 a . likewise , tabs of configuration 46 a are inserted into complementary grooves of configuration 45 a . each side of exercise platform 47 also comprises at least one slot 47 a . during assembly , slot 47 a receives at least one tab 46 b of side wall 46 or at least one tab 45 a of indicia wall 45 . the components of box assembly 40 b are thereby strongly interconnected . in various embodiments the components of box assembly 40 b may be connected by screws , bolts , nails , or adhesives , interlocking components or by any other mechanical means known in the art . fig1 a , 16 b , and 16 c show a first side view of base assembly 40 a , second side view of base assembly 40 a rotated through 90 degrees from the previous view , and isometric overview of base assembly 40 a , respectively . fig1 a , 17 b , and 17 c show a first side view of box assembly 40 b , second side view of box assembly 40 b rotated through 90 degrees from the previous view , and isometric underview of box assembly 40 b , respectively . plyometrics exercise assembly 40 is preferably constructed of engineered wood , such as high - density plywood . it can be seen through the description of this invention that various alternative embodiments are possible without deviating from the scope and spirit of this invention .

the present invention is a telescoping plyometric exercise device which can be rapidly adjusted for height using a single lifting motion . the apparatus is incrementally height - adjusted and secured in a position capable of withstanding force . the apparatus requires a single lifting motion to securely and incrementally adjust the device , and to balance impact and force through the use of novel , contoured inner components and structural assemblies . these inner components and structural assemblies are engineered to provide the capability for rapid height adjustment of the apparatus .

a receptacle 10 for use with this invention has a wall 11 which forms an internal cavity 12 . a port 13 through the top of the container has an externally threaded neck 14 to receive a cap 15 . it is preferred that cap 15 be of a typical ratchet - type child - safe type which is intended to limit access , and to be less likely to loosen in the event the container is knocked over . this safety feature is optional . a rigid locking ring 16 , shown separately in fig3 is kept available but in a place where it is unlikely to be attached to the receptacle until the continuing use of this receptacle is concluded . at such time , it will be placed around the neck between the cap and the receptacle when a permanent closure is required . it is not compressible and will prevent the cap from being depressed after the cap is tightened . then the ratchet cannot thereafter engage and the cap cannot be unthreaded from the neck . a suitable quantity of dry solid material 20 is added to the receptacle , into which the sharps will be placed . the level to which the solid material is added leaves sufficient open volume even after all of the sharps are placed in the receptacle , that the sharps and a slurry to be described can be received and shaken together with the solid material and sharps . the presently preferred solid material is dry calcium hypochlorite -- ca ( ocl ) 2 in granule or powder condition . sharps can be dropped into the container up to a sharps fill line 21 on the wall of the receptacle . a supply container 25 has a wall which forms an internal cavity 26 to store a desired amount of the solid reagents and materials 27 needed for the final solidification , encapsulation and disposition of the sharps . it has a port 28 with an externally threaded neck 29 . neck 29 receives a removable cap 30 that forms a closure to contain the materials . a reference line 31 is provided to show the level of water to be added , and still leave room for shaking the water and solid material to form a slurry . according to this invention the materials in the supply container are kept dry until they are to be used . they are for the purpose of reducing any remaining hypochlorite in the receptacle , for creating a slurry with a ph above 11 . 5 , for forming a cementitious body encapsulating the sharps , and for reducing the temperature generated by the curing of the cements and by reduction of the hypochlorite . the receptacle and the supply container are sold as a kit so the substances in them will be present when the need for them arises . the materials in the supply container are : portland cement and pumice are provided to form a cementitious body . potassium chloride is provided for temperature reduction . all are provided for creating a slurry with a ph above 11 . 5 . the use of this product is straightforward . sharps are dropped into the receptacle , and the cap is replaced after each insertion . a very faint bleach odor will be detected , which is neither dangerous nor objectionable . because chlorine is a heavier gas , it is unlikely to escape the receptacle , and likely to be retained in it . it is a &# 34 ; friendly &# 34 ; smell , and indicates that chlorine is present to destroy the microorganisms , which in fact it does . when sufficient sharps have been inserted into the receptacle , or when the desired maximum shelf life of the hypochlorite has passed , the supply container and its contents will be used . the half life of the hypochlorite in the receptacle is about 2 years . a sensible shelf life is about one year . in the supply container , water is added to line 31 , and the container is closed and shaken vigorously for about thirty seconds . this will activate these materials and form a slurry . this slurry is promptly poured from the supply container into the receptacle . it is closed , and is vigorously shaken for about thirty seconds to thoroughly mix the contents . the supply container can be separately disposed of as ordinary trash . now , in addition to the setting of the cement in the receptacle , the following reaction occurs : this is in addition to the curing of the cement and the formation of the matrix with the pumice . the potassium chloride takes no part in the reaction . when used , and it need not always be used , it merely accepts calories with its negative heat of solution to keep the temperature at a maximum of about 125 degree f . which is best not to exceed so that the integrity and safe handling of the plastic container will not be compromised . after about 48 hours , the mass in the receptacle will have been set and cured . the ring 16 will have been placed around the receptacle neck , and the receptacle cap will then have been closed . the receptacle , now an urn , is permanently closed and ready to go to the landfill for permanent disposal . this material qualifies for long term disposal in a non - hazardous landfill . the material of the receptacle itself is preferably non - biodegradable , impermeable and strong , for example high density polypropylene . the supply container may be an ordinary polyethylene bottle . its disposal represents no problem . a convenient volume for the receptacle is about 2 quarts . about 50 gm of calcium hypochlorite will be added , which after the sharps are inserted to capacity , leaves a volume of about 1 . 5 quarts as open volume in which to receive the slurry . a convenient volume for the supply container is about 1 gallon . it will be filled about as follow : ______________________________________sodium sulfite : 70 - 90 preferably 79 gmpotassium chloride : 150 - 250 preferably 200 gmpumice : 500 - 600 gm preferably 570 gmportland cement : 800 - 1 , 000 gm preferably 920 gm______________________________________ about 1 , 300 ml ( 37 fl oz ) of water will be added to form the slurry in the preferred amounts . these amounts may of course be varied , but have been found to be optimal for the purpose intended . the sodium sulfite is provided in an approximately stoichiometric amount , even though it may be in excess after the calcium hypochlorite has previously degraded or reacted . it is best practice to be certain that all of it has been reacted . other advantages of this invention become evident with its use . many sharps have hollow cavities or passages , such as needles and barrels . it is most desirable for these to be filled , even though they are sometimes of near capillary dimensions . in fact the cementitious material expands as it starts to set . it tends to flow readily into the spaces and lumens , assisted by the forces of capillary action and of expansion of the mass . this is a secondary and useful functional feature . furthermore , the alkalinity of this composition further inhibits any organisms which might somehow have survived the hypochlorite . this invention is not to be limited by the embodiment shown in the drawings and described in the description , which is given by way of example and not of limitation , but only in accordance with the scope of the appended claims .

apparatus and method for the interim sanitary storage of medical sharps and subsequent inurnment of them to a condition acceptable in commercial landfills . a receptacle contains dry calcium hypochlorite to sanitize sharps that are placed in the reception . it has a removable closure cap to admit the sharps . a supply container contains cementitious material to which water is added , and then poured into the receptacle to envelop the sharps . a permanent lock is applied to the receptacle cap to prevent further access or escape of the sharps which are encapsulated in the resulting cementitious urn .

it must be understood that , just as the human - visible colors present on an animal vary greatly over the surface of that animal , the ultraviolet ( uv ) light reflected from the surface of that animal can also vary greatly . a human - visible color such as the green found on the head of a drake mallard duck is simply a set of reflected wavelengths of light that falls within the spectrum of light visible to humans ; specifically , it is wavelengths of light that humans perceive as the color green ( for illustration purposes only we ignore the iridescent “ angle - dependant ” nature of these feathers ). depending on the exact wavelength and intensity of the reflected light , the color “ green ” may range in appearance from blue - green to yellow - green . similarly , the amount of uv light , as well as the specific wavelengths of uv light , reflected from the surface of an animal can vary greatly , creating different “ colors ” of uv light . although these uv colors are not visible to humans , they are visible to many animals , and should be accounted for when creating realistic models or decoys of those animals . that is the intent of the present invention . throughout this specification and in the claims , the terms “ matches ” or “ substantially matches ” and derivations thereof are used to describe to what degree the surface reflection patterns of the decoy and decoy lures created through the employment of this invention will appear natural to the animals for which they are intended to deceive . it is possible , for example , for someone to increase the total amount of ultraviolet light reflected from a decoy or decoy lure by covering it with an ultraviolet - reflecting material such as tyvek for bird decoys or embossed foils for decoy lures . while there is an overall increase in the amount of ultraviolet light reflected from such a decoy , this decoy will not “ substantially match ” the varied surface refection pattern of a real animal or object , and will not appear realistic to the target animal . the terms “ matches ” and “ substantially matches ” and derivations thereof , as used in this specification , are used to describe decoys and decoy lures with surface reflectance patterns which will appear realistic to the target animal , and which fall within the range of ultraviolet wavelengths seen in nature for the corresponding portions of the actual target animal or object being mimicked . fig1 illustrates the uv reflectance patterns of two different waterfowl . although waterfowl are used as examples herein , it should be understood that any type of animal can be used with similar results . fig1 shows areas of high uv reflectance 10 , areas of medium uv reflectance 20 , and areas of little or no uv reflectance 30 in the patterns in which they would typically appear on a drake mallard duck or a canada goose . although the present invention shows areas of high uv reflectance 10 are often seen associated with areas of white or light human - visible colors on the waterfowl , studies have shown that this is often not the case for all white colored animals . similar studies have shown that areas of black can be associated with significant uv reflectance , and areas of white can have very little uv reflectance . white feathers and many other white surfaces on animals ( for example some white fish scales ) often possess uv reflection of 50 to 70 % ( a common range of waterfowl white feathers and some white fish scales ) but could be as high as 90 % and as low as 15 %. these percentages represent averages of the reflectance across the uv wavelengths visible to the targeted animal . similarly , non - white colors have ranges typical for each species . these percentages can be determined using uv - visible reflectance curves . the uv - visible reflectance curves of some common decoy and decoy lure colors are shown in fig2 . fig2 illustrates the uv - visible reflectance curves of example animals as compared to materials found in the prior art used for coating decoys or decoy lures . the visual spectrum of humans 60 and the visual spectrum typical of birds , many fish and other animals 61 are indicated along the bottom access of the line graph . the reflectance curves of several materials taught in the prior art , including white 40 , light tan 41 , and tan 42 , are shown . each of the materials 40 , 41 , and 42 demonstrates very little reflectance in the wavelengths of ultraviolet light between 300 and 400 nanometers . the reflectance curves of a snow goose body and wing 50 , a canada goose cheek patch 52 , a canada goose breast 54 and a typical white area on a bait fish such as a minnow 97 are also shown . the reflective characteristics of animal components 50 , 52 , 54 , and 97 cannot adequately be implemented using materials 40 , 41 , and 42 . animals which can see in the visual spectrum of birds and many fish 61 will see materials 40 , 41 , and 42 as significantly different colors than animal components 50 , 52 , 54 , and 97 even though materials 40 , 41 , and 42 will appear as close matches in the human visible spectrum 60 . the range of uv wavelengths within the visual spectrum of animals 61 , can vary from species to species . this range could be as wide as 300 to 400 nm but is more commonly about 320 to 400 nm . this effective visible uv range can be about 350 to 400 nm for some species such as many waterfowl or as narrow as about 380 to 400 nm . the present invention describes a method of mapping the reflectance characteristics of the outer surface of an animal . refer now to fig3 and fig4 . fig3 illustrates a test set - up which uses uv imaging or similar techniques to determine areas of low , medium , and high reflectance on the outer surface of the animal . the animal subject 70 is placed in front of a uv imaging camera 74 . light sources 72 emit ultraviolet light onto the animal subject 70 , and the reflected uv light is detected by the uv imaging camera 74 . a monochrome image 78 , showing areas of high uv reflectance as bright areas , moderate uv reflectance as shades of gray and no uv reflectance as dark areas , is displayed on a computer display 76 . the data from the image 78 is interpreted and recorded to show a map of the uv reflection pattern like that shown in fig1 . fig4 illustrates an additional step in which the animal subject 70 is mapped with a uv - vis spectrophotometer 80 to determine the quantitative reflectance curves 84 across the spectrum of the animal vision system . surface measurements are taken from whole or partial samples 82 from carcasses or other natural samples . a reflectance curve 84 is generated in this manner for each different sample 82 . example reflectance curves 84 can be seen in greater detail for the animal components 50 , 52 , 54 and 97 on fig2 . fig5 is a flowchart of the process of creating animal decoys exhibiting realistic uv reflections . in step 90 , a uv reflectance surface map is created for the animal subject 70 . this is done by the uv imaging process previously described herein in fig3 . in step 91 , uv reflectance curves 84 are created for various samples 82 of an animal carcass . this is done by the uv - vis process previously described herein in fig4 . the uv image 78 and reflectance curves 84 are analyzed to create specific formulations of paint or other surface covering material , as shown in step 92 . in step 93 , the uv - reflective paint or material is applied to the outer surface of an animal decoy , or the decoy itself is composed of said materials , to create a model of the animal subject 70 that appears visually realistic to the animal in the animal &# 39 ; s visual spectrum . fig6 illustrates the uv general reflectance patterns of common bait fish minnows such as flathead , shiners , suckers and many other common bait fish . the dark top 94 is typically brown , tan , and black patterns and contains a low level of uv reflection , typically less than 20 %. the silvery sides 95 possess bright specular uv reflectance , as much as 80 % or higher at certain angles . the belly 96 possess bright diffuse uv reflectance from 15 - 80 %. current lures attempt to achieve this diffuse reflectance with conventional paints and as such are uv - absorbing , typically about 90 % uv absorbing ( or more ) or only 10 % reflecting ( or less ) in the uv wavelengths . the present invention matches the diffuse uv reflection of animals such as bait fish which is typically 15 % to 80 % uv in the areas described in fig6 . while it is apparent that some of the current art inadvertently often achieves some specular ( non - diffuse ) uv reflection of the sides 95 of a bait fish with the use of embossed or otherwise textured metal foils , it is also apparent the diffusely reflecting surface is achieved with paint , coatings , printing inks , or molded - in colored surfaces which are typically plastic , and as such will be uv - absorbing as said surfaces employ tio2 to achieve diffuse reflectance of white and light colors . in the preferred embodiment , one or more surfaces of a decoy or decoy lure are coated or coloration is molded - in to achieve diffuse uv reflection on those surfaces where diffuse uv reflection is present in the animal to be mimicked . this diffuse reflection is typically between 15 to 80 % uv reflecting in the wavelengths from 320 nm to 400 nm with the preferred embodiment being 30 % to 70 % for human - visible white ( and other light colors and some bright colors ) but could be as low as 12 % and as high as 90 %. in another embodiment , the underside or belly areas of a decoy lure are painted with diffusely reflecting uv paint which is human - visible white , or light - colored , or bright - colored but it will become apparent that this pattern of uv reflection applies to certain bait fish , and other animals to be mimicked by decoys or decoy lures could have different diffusely uv - reflective areas . the methods used to measure this uv reflectance and subsequently to develop coatings or molded - in colors are established in u . s . patent application ser . nos . 11 / 711 , 409 and 11 / 711 , 410 , to which this application is a continuation in part . the ability to discriminate and measure diffuse versus specular reflectance is a common attribute of uv / vis spectrophotometers and understood by those skilled in the art . having described the preferred embodiment and other example embodiments , it will become apparent that various modifications can be made without departing from the scope of the invention as defined in the accompanying claims . in particular , any animal , plant , seeds , or even an object , can be used as a decoy or decoy lure if it aids in the deception of an animal . the common waterfowl and bait fish minnow discussed directly herein are intended as examples only and the teachings of this application could be applied to any object intended to deceive or lure an animal . in addition , methods of measuring the uv reflectance of an animal or object other than those discussed herein may be used to achieve the same or similar results . using the methods described , it will also become apparent that certain animals may have cryptic uv reflecting surfaces that do not correspond to bright human visible colors . it is obvious that many animals vary in human - visible appearance between male and female ; using the methods described , it will become apparent that game animals and bait fish may possess human - invisible appearance differences that could be mimicked to make decoys or decoy lures more effective .

an animal decoy or fishing lure with a surface reflection which closely matches the spectral reflectance pattern of the animal that it is designed to mimic , including both human - visible and ultraviolet wavelengths , with the intent of making the decoy or fishing lure appear more realistic to animals that can see in both the human - visible and ultraviolet spectrums .

reference will now be made in detail to embodiments of the present invention , examples of which are illustrated in the accompanying drawings , wherein like reference numerals refer to the like elements throughout . the embodiments are described below in order to explain the present invention by referring to the figures . fig1 is a perspective view illustrating an accessory according to an embodiment of the present invention , fig2 is a partial enlarged front view illustrating internal elements of the accessory of fig1 , and fig3 is a partial enlarged cross - sectional view illustrating the internal elements of the accessory of fig1 . referring to fig1 through 3 , accessory 100 is a kind of necklace . accordingly , the accessory 100 comprises an accessory body 110 in the shape of a necklace , a measurement unit 120 mounted on the accessory body 110 , an ic chip 132 including a signal processing circuit , a monopole antenna 142 , and a ground 144 . a part of the measurement unit 120 is exposed outward to be capable of maintaining contact with the body of the user directly or indirectly . the accessory body 110 is in the form of a general necklace and includes a pendant 112 and a necklace string 114 . the measurement unit 120 that can be attached to the body is mounted on the rear side of the pendant 112 , the ic chip 132 and the monopole antenna 142 are mounted on a printed circuit board ( pcb ) substrate 130 in the pendant 112 . the top of the pendant 112 is connected to the necklace string 114 , and the monopole antenna 142 and the ground 144 are electrically connected via the engaged parts of the pendant 112 and the necklace string 114 . the ground 144 is built into and along the necklace string 114 and is formed in a length of more than approximately λ / 4 , where the symbol of λ indicates a wavelength of a used frequency . accordingly , the ic chip 132 may form a wireless communication unit by using the monopole antenna 142 , and the ground 144 may function as a valid image antenna with respect to the monopole antenna 142 . the measurement unit 120 installed in the pendant 112 may receive a signal for monitoring a biosignal of the user by being in contact with the body of the user . generally , the biosignal may include an electrocardiogram , an electromyogram , an electroencephalogram , and a galvanic skin response . the measurement unit may be also used for measuring a body temperature , a pulse , or a blood pressure . in addition , the measurement unit 120 may further include other functions . in addition to the use of measuring a change in the body of a user , for example , a temperature , a pressure , a humidity , a components of air , a composition of air , a radioactivity , and a deleterious bacteria analysis of the surrounding environment may be measured . to measure environmental characteristics instead of physiological characteristics , the measurement unit may be mounted on the front of a pendant instead of the rear of the pendant . in the present embodiment , though the measurement unit 120 includes one contact terminal , according to the type and number of biosignals to be measured , two or more contact terminals may be provided and positions of contact terminals may be determined . when a plurality of contact terminals are formed , each of the contact terminals is independently connected to the signal processing unit , and the signal processing unit may generate a signal to be transmitted to the wireless communication unit via a process of amplifying , filtering , or combining signals transmitted from each of the terminals . fig4 is a partial enlarged front view illustrating internal elements of a pendant of the accessory of fig1 , and fig5 is a diagram illustrating function transfer between the internal elements of fig4 . referring to fig1 - 5 , the measurement unit 120 disposed in the rear side of the pendant may be in continuous contact with the body of the user and may measure electrocardiogram or electromyogram . a signal measured by the measurement unit 120 may be an analog signal . the ic chip 132 , as a signal processing unit , receives the analog signal from the measurement unit 120 and may process the received signal to transmit to an external apparatus or server . generally , the ic chip 132 may amplify , filter , and convert the measured signal into a digital signal , and the processed signal may be transmitted to an external apparatus or server via the wireless communication unit 140 . in the present embodiment , it is described that the accessory 100 measures a biosignal and transmits the biosignal to the external apparatus or server . however , the accessory 100 may receive emergency information from the external apparatus or server and may display the degree of the danger for giving a warning to the user . namely , the wireless communication unit 140 may include not only a function of transmission but also a function of reception . fig6 is a perspective view illustrating an accessory 200 according to another embodiment of the present invention . for reference , the accessory 200 of fig6 is a necklace but includes a measurement unit and a ground mounted separately from a pendant . referring to fig6 , the accessory 200 includes an accessory body 210 formed in the shape of a necklace , a measurement unit 220 provided on the side of a string of a necklace and in contact with a body of a user , a signal processing unit 230 mounted on the measurement unit 220 , and a ground 244 . the measurement unit 220 is formed in the shape of a disk , as a conventional sensing electrode for medical use . an electrode in contact with the user &# 39 ; s body is included underneath the measurement unit 220 . the signal processing unit 230 is mounted on the top of the measurement unit 220 and includes an ic chip including a signal processing circuit and a monopole antenna . the ic chip of the signal processing unit 230 measures , converts , and processes a biosignal . the ic chip is electrically connected to the electrode of the measurement unit 220 . also , the monopole antenna of the signal processing unit 230 is functionally connected to the ground 244 . the measurement unit 220 and the signal processing unit 230 are installed in a part of the string of the accessory body 210 , and the ground 244 in the body . though the measurement unit 120 and the pcb substrate 130 are built into the accessory body 110 in fig1 and 2 , in the present embodiment , the measurement unit 220 and the signal processing unit 230 are formed on the outside of the accessory body 210 and the ground 244 is wired by using the string . the ground 244 is formed in a length greater than approximately λ / 4 of a used frequency . accordingly , the ic chip of the signal processing unit 230 may form a wireless communication unit by using the monopole antenna and the ground 244 , and the ground 244 may function as a valid image antenna with respect to the monopole antenna . the measurement unit 220 mounted on the string may receive a signal for monitoring a biosignal of a user by being in contact with the user &# 39 ; s body . as described above , the biosignal may include an electrocardiogram ( ecg ), an electromyogram ( emg ), an electroencephalogram ( eeg ), a galvanic skin response ( gsr ), a body temperature , a pulse , and a blood pressure . in the present embodiment , one measurement unit 220 is mounted on the string , but the type or number of measurement units may vary . when a plurality of measurement units is formed , the measurement units may be independent or may be dependently connected to each other along the string . the measurement 220 may measure an ecg or an emg while being in contact with the user &# 39 ; s body . the signals measured by the measurement unit 220 may be prepared to be transmitted externally via the ic chip of the signal processing unit 230 . the ic chip may amplify , filter , and convert the measured signal into a digital signal to transmit to an external apparatus or server via a wireless communication unit . fig7 is a perspective view illustrating an accessory 300 according to still another embodiment of the present invention . referring to fig7 , the accessory 300 is a watch and includes an accessory body 310 in the shape of a watch , a measurement unit 320 mounted on a part of the accessory body 310 and in contact with a user &# 39 ; s wrist , a signal processing unit 330 including an ic chip including a signal processing circuit , a monopole antenna ( not shown ), and a ground 344 . the accessory body 310 includes a watch body and a watch band 314 . the measurement unit 320 capable of being in contact with the wrist is installed in the bottom of the watch body , and the signal processing unit 330 including the ic chip is formed in a watch body together with the measurement unit 320 . also , the signal processing unit 330 includes the ic chip and the monopole antenna on a single pcb substrate . since internal elements of the accessory have been previously described with reference to fig4 , detailed configuration and description will be omitted . referring to fig7 , both sides of the watch body are connected to the watch band 314 , and the monopole antenna and the ground 344 are electrically connected to a portion of the connection between the watch body and the watch band 314 . the ground 344 is built in the watch band 314 and formed in a length of at least more than approximately λ / 4 of a used frequency . accordingly , the ic chip may form a wireless communication unit by using the monopole antenna and the ground 344 , and the ground 344 may function as a valid image antenna with respect to the monopole antenna . the measurement unit 320 mounted on the watch body may receive a signal for monitoring a biosignal of a user while being in contact with the user &# 39 ; s wrist . generally , the biosignal may include an ecg , an emg , an eeg , a gsr , a body temperature , a pulse , and a blood pressure . in addition , the measurement unit may sense a temperature , a pressure , a humidity , a components of air , a composition of air , a radioactivity , and a deleterious bacteria of the surrounding environment . in the present embodiment , the measurement unit 320 includes one contact terminal , but according to the type and number of biosignals to be measured , two or more contact terminal may be provided and a position of the contact terminal may be determined . the measurement unit 320 in the rear of the watch body may measure an ecg or an emg while being in contact with the user &# 39 ; s wrist . a signal measured by the measurement unit 320 is an analog signal . the ic chip of the signal processing unit 330 receives the analog signal from the measurement unit 320 and may prepare to transmit the received signal outside by processing . the ic chip may amplify , filter , and convert the measured signal into a digital signal , and the processed signal may be transmitted to an external apparatus or server via a wireless communication unit . fig8 is a perspective view illustrating an accessory 400 according to yet another embodiment of the present invention . referring to fig8 , the accessory 400 is motivated by glasses and includes an accessory body 410 in the shape of glasses , a measurement unit 420 mounted on a part of the accessory body 410 and in contact with sides of a user &# 39 ; s head , a signal processing unit 430 including an ic chip including a signal processing circuit , a monopole antenna ( not shown ) and a ground 444 mounted together with the ic chip on a substrate . the accessory body 410 includes a glasses body and glasses legs ( or temples ) 414 formed on both ends of the glasses body . the measurement unit 420 capable of being in contact with a side of a face is installed in one of the glasses legs 414 . the signal processing unit 430 including the ic chip is installed in the glasses leg 414 , close to the measurement unit 420 . also , the signal processing unit 430 includes the ic chip and the monopole antenna on a single pcb substrate . the measurement unit 420 , the signal processing unit 430 , and the ground 444 are sequentially provided along the glasses leg 414 . the ground 444 is built in the glasses leg 414 and formed in a length greater than approximately λ / 4 of a used frequency . accordingly , the ic chip may form a wireless communication unit by using the monopole antenna and the ground 444 , and the ground 444 may function as a valid image antenna with respect to the monopole antenna . the measurement unit 420 mounted on the glasses leg 414 may receive signals associated with an ecg , an emg , an eeg , a gsr , a body temperature , a pulse , and a blood pressure of a user while being in contact with the head . though the measurement unit 420 is mounted in one of the glasses legs 414 in the present embodiment , according to the type and number of biosignals to be measured , two or more measurement units may be provided in each glasses leg . the measurement unit 420 in the glasses leg 414 may measure a desired biosignal while being in contact with the head . a signal measured by the measurement unit 420 is processed via the ic chip of the signal processing unit 430 , and the processed signal may be transmitted to an external apparatus or server by using the monopole antenna and the ground 444 . the accessories of the above - described embodiments of the present invention may be conventional accessories or small articles and may include a micro circuit unit and an antenna . however , since a ground has to have a certain length for effective communication , the ground may be built or wired into an accessory body in the certain length . a ground having more than the certain length makes an efficient wireless communication of the accessory possible . since the shapes of conventional clothing articles or accessory can be used as an accessory body of the present invention , it is simple to put on or carry , thereby maximizing convenience of users . although a few embodiments of the present invention have been shown and described , the present invention is not limited to the described embodiments . instead , it would be appreciated by those skilled in the art that changes may be made to these embodiments without departing from the principles and spirit of the invention , the scope of which is defined by the claims and their equivalents .

an accessory for remote monitoring , including : a body capable of being placed on a human body ; a measurement unit installed in the body and measuring a change in surroundings ; a signal processing unit for processing a signal acquired from the measurement unit ; and a wireless communication unit including a monopole antenna transmitting the signal processed by the signal processing unit and a ground installed along the body , corresponding to the monopole antenna . the signal processing unit and the monopole antenna are manufactured in a small size to be formed in the accessories body or periphery of the accessories body , and a space for wiring the ground may be obtained by using a band or the accessories body in an extended shape .

the organoarsenic imaging agents of the present invention are composed of three main parts . the receptor binding / recognition domain of the compounds will be either a peptide or a small organic molecule . in either case , the molecule possesses known receptor specificity . in other words , the peptide or small molecule has a documented affinity and favorable binding kinetics at a receptor site in a tissue of interest . regarding breast cancer , the drugs tamoxifen and diethylstilbesterol ( des ) both fit these requirements . this portion of the imaging compounds will be used as the delivery agent as well as the receptor recognition and binding domain . the introduction of the spacer domain to the prepared agents is also of high importance to binding . the binding characteristics of the receptor recognition and binding domain are dependent upon the size and geometry as well as hydrophilicity of the molecule . if the geometry of the ligand is distorted or its dimensions within the binding confines of the receptor made too large , the ligand - receptor complex will not form and no tissue specificity will be imparted . since arsenic has an average single bond colvalent atomic radius of 121 pm ( roughly 1 . 6 times larger than carbon ), it is necessary to make sure this atom is well removed from potentially devastating interactions with receptor . the function of the spacer domain is to tether the arsenic to the receptor recognition ligand while not allowing the large atomic size of the metal to prevent binding to the receptor . this allows specific delivery of the radioactive arsenic to the tissue receptor of interest for imaging . for suitable peptide ligands , the peptide will be coupled to the spacer and the spacer to a lipoic acid appendage using dicyclohexylcarbodiimide ( dcc ) chemistry . this technique is the standard for peptide preparation and derivatization within the chemical industry . next , the reduction of the disulfide bond in the lipoic acid to the dithiol will be accomplished . mild conditions must be used so as to not affect any other portion of the peptide . though numerous mild techniques for this type of reduction exist , preferred is the removal of a s , s &# 39 ;- isopropylidene protecting group with mercury ( ii ) chloride and aqueous mercury ( ii ) acetate cleavage of a protected s - phenylthiomethyl dithioacetal . once the dithiol is formed , addition of as + 3 will result in formation of a covalently bound arsenic through two sulfur - arsenic bonds ( see below ). this will be done first using &# 34 ; cold &# 34 ; ( a non - radioactive isotope ) arsenic and then , following satisfactory results , 72 as will be incorporated . ## str2 ## the detection , purification and characterization of the prepared compounds will involve standard reverse - phase high performance liquid chromatography ( hplc ). ultraviolet as well as radiochemical detection methods will be employed . following purification , the compound ( s ) will be ready for binding and specificity assessment . the modification of either steroids or steroid surrogates provides an opportunity to identify cellular populations with high densities of a receptor . the estrogen receptor is implicated in breast cancer and the use of antiestrogens in the treatment of estrogen dependent tumors is recognized as important therapeutically . this strategy is less attractive than modification of peptides due to the relatively large change in the physical characteristics of the compound with the incorporation of a functional group capable of chelation of arsenic . specifically ethers of estrogens , which contain a lipoic acid - like group , are prepared for the chelation of arsenic as shown in the figure below . ## str3 ## from the foregoing , it will be appreciated that , although specific embodiments of the invention have been described herein for the purposes of illustration , various modifications may be made without deviating from the spirit and scope of the invention . accordingly , the invention is not limited by the appended claims .

radioactive arsenic labelled tissue specific ligands are prepared and used in the medical imaging of malignant breast or prostate tumors .

a ) a particulate component that is difficult to dissolve in water , and , b ) at least one liquid or gel component that is soluble in water , the hydrogel component that dissolves more easily acts as an intrinsic pore former in the bone defect . the intermediate spaces between the particles of the preparation , the size of which is a function of the particle size and the particle size distribution , promote the ingrowth of veins and tissue . the hydrogel matrix that dissolves more easily also expands the agglomeration of the particles and simultaneously promotes cohesion in order to assure injectability . as fig1 depicts , the hydrogel matrix ( 1 ) is located between the particles ( 2 ). when the inventive preparation is inserted into the defect , blood ( 3 ) coagulates , calcium - indicated , on the surface of the agglomeration of the particles , which is depicted schematically in fig2 . the hydrogel component that dissolves more easily is resorbed within a short period of time so that the particle agglomeration stabilized by fibrous tissue remains . this agglomeration made of resorbable bioceramic having large interparticulate intermediate spaces now satisfies all requirements for a bone replacement material in terms of porosity , freedom from phase shift , and resorbability . the following is understood for the term not easily soluble . the not easily soluble substance dissolves in water at room temperature in a maximum concentration of 100 mg / l . in accordance with the invention , the particles in the preparation have a size in the range from 0 . 1 to 150 μm , pores having a diameter of 0 . 01 - 50 μm , and can comprise β - tricalcium phosphate , α - tricalcium phosphate , whitlockite , octacalcium phosphate , hydroxyapatite , type a carbonate apatite , type b carbonate apatite , calcium - deficient hydroxyapatite , amorphous calcium phosphate , and / or resorbable glass ceramics . the calcium phosphate particles can have both a round shape and a polygonally broken shape that is rounded by wear and thermal sintering . the aforesaid particles in the size range of 0 . 1 to 150 μm have pore diameters of 0 . 01 to 50 μm . the inventive , plastically deformable preparation can be paste - like or can have a very viscous , more kneadable to wax - like form . the paste - like form of the preparation is designed such that it can be applied to a bone defect in a minimally invasive manner by means of an injection syringe having a straight or angled cannula . the diameters of the particles are optimized for the cannula diameter of an injection cannula . particle diameters are optimized such that the paste - like material can also be applied using an cannula that is angled up to 60 °, measured with respect to the longitudinal axis of the injection syringe . it is essential for the invention that it is constructed from a mixture of 60 - 90 % by mass calcium phosphate particles and 1 - 30 % by mass of an aqueous or alcoholic solution of dextran and / or carboxymethyl dextran and / or hyaluronic acid and / or dermatan sulfate , carboxymethyl cellulose and / or oxidized cellulose and / or gelatins and / or mixtures thereof . the use of polysaccharide derivatives , such as for instance carboxymethyl dextran , carboxymethyl hyaluronic acid , and sulfated hyaluronic acid , is also within the sense of the invention . it is likewise inventive that it is preferably constructed from a mixture of 80 - 98 % by mass calcium phosphate particles , 1 - 20 % by mass anhydrous polyethylene glycol 400 , and 1 - 20 % by mass anhydrous polyethylene glycol 600 . polyethylene glycol 400 and polyethylene glycol 600 can also contain oxidation stabilizers , in addition . fig3 provides detailed images of the x - ray powder diffractograms of ( 1 ) the initial batch of a β - tcp ceramic ( 2 ) β - tcp in hyaluronic acid solution ( 3 ) β - tcp in dextran solution ( 4 ) β - tcp in methylcellulose solution after 12 weeks &# 39 ; storage in water . as can be seen from this documentation , in the inventive formulation there is no conversion to a different phase of the calcium phosphate family . fig4 provides detailed images of the x - ray powder diffractograms of ( 1 ) the initial batch of a β - tcp ceramic ( 2 ) the same ceramic after 12 weeks &# 39 ; storage in water as can be seen from this documentation , in the aqueous suspension there was partial conversion / recrystallization of β - tcp to hydroxyapatite . the inventive preparation can be sterilized with dry heat or using gamma rays at a dosage of 8 to 30 kgy (= kilograys ). the hyaluronic acid used for the inventive preparation or the hyaluronic acid salts can be bioengineered , the hyaluronic acid having a molecular weight of 1 , 500 , 000 to 4 , 500 , 000 daltons and sterilization reducing this to 700 , 000 to 2 , 500 , 000 daltons . the invention shall be explained using examples in the following , but the invention shall not be limited to these examples . components a through k can be used for producing the plastically deformable implant material . 99 % β - tricalcium phosphate , free of phase shift , polygonally broken , the broken edges rounded by abrasion and subsequent burning , having a porosity of 20 ± 5 % and particle size distribution of 0 . 1 - 50 μm , bulk density of 1 . 1 ± 0 . 1 g / cm 3 , and grain size of & lt ; 63 μm ( d 50 = 15 ± 5 μm ) 0 . 3 g component a are added to 20 . 05 g component c . after a 4 - hour swelling time , the gel is sterile - filtered , slowly added to a total of 50 g component b , and mixed intimately . the mixture is sterilized in a container at 121 ° c . using dry heat . the result is a homogenous mixture with a paste - like consistency that is easily extruded through a cannula , conforms to a defect due to plastic flow , and that air easily hardens superficially , it being possible to accelerate this process using a gentle airflow from a compressed air pistol . after heat sterilization the material has a molecular weight of 1 . 7 ± 0 . 5 mda . even after 4 hours of centrifuging , no solid or liquid settles out . an extruded paste adheres well to surrounding tissue . an extruded and superficially dried body retains its shape in the simulated body environment ( moist chamber under physiological conditions ). 0 . 2 g component a are added to 20 . 0 g component c . after a 4 - hour swelling time , the swollen gel is sterile - filtered , slowly added to a total of 63 . 4 g component b while stirring , and mixed intimately . the mixture is sterilized in a closed container at 121 ° c . using dry heat . the result is a homogenous mixture that has a plastic consistency and is kneadable , that because of its kneadability is easily applied to a bone defect , that adheres well to the surrounding tissue , and that air easily hardens superficially , it being possible to accelerate this process using a gentle airflow from a compressed air pistol . after sterilization the material has a molecular weight of 1 . 7 ± 0 . 5 mda . even after 4 hours of centrifuging , no solid or liquid settles out . a hand - formed and superficially dried body retains its shape in the simulated body environment ( moist chamber under physiological conditions ). 225 mg component d are added to 38 mg component a , and then 9 . 7 g component c is added while stirring . after a 1 - hour swelling time , the swollen gel is sterile - filtered , slowly added to a total of 22 g component b while stirring , and mixed intimately . the mixture is sterilized in a closed container at 121 ° c . using dry heat . the result is a homogenous mixture with a paste - like consistency that is easily extruded through a cannula , conforms to a defect due to plastic flow , and that air easily hardens superficially , it being possible to accelerate this process using a gentle airflow from a compressed air pistol . even after 4 hours of centrifuging , no solid or liquid settles out . a large quantity of the material does not result in an aqueous solution settling on the surface , even after standing for a number of weeks . an extruded paste adheres well to surrounding tissue . an extruded and superficially dried body retains its shape in the simulated body environment ( moist chamber under physiological conditions ). 200 mg component e are mixed with 100 mg component a , and then 19 g component c are added while stirring . after a 1 - hour swelling time , the swollen gel is sterile - filtered , slowly added to a total of 58 g component b while stirring , and mixed intimately . the mixture is sterilized in a closed container at 121 ° c . using dry heat . the result is a homogenous mixture that has a plastic consistency and is kneadable , that because of its kneadability is easily applied to a bone defect , that adheres well to the surrounding tissue , and that air easily hardens superficially , it being possible to accelerate this process using a gentle airflow from a compressed air pistol . even after 4 hours of centrifuging , no solid or liquid settles out . a large quantity of the material does not result in an aqueous solution settling on the surface , even after standing for a number of weeks . a hand - formed and superficially dried body retains its shape in the simulated body environment ( moist chamber under physiological conditions ). 500 mg component f are mixed with component c to make 10 . 0 g . after a 4 - hour swelling time , the swollen gel is sterile - filtered , slowly added to a total of 27 g component b while stirring , and mixed intimately . the mixture is sterilized in a closed container at 121 ° c . using dry heat . the result is a homogenous , plastic mass that is kneadable , that because of its kneadability is easily applied to a bone defect , that adheres well to the surrounding tissue , and that air easily hardens superficially , it being possible to accelerate this process using a gentle airflow from a compressed air pistol . even after 4 hours of centrifuging , no solid or liquid settles out . a hand - formed and superficially dried body retains its shape in the simulated body environment ( moist chamber under physiological conditions ). 500 mg component g are mixed with component c to make 10 . 0 g . after a 4 - hour swelling time , the swollen gel is sterile - filtered , slowly added to a total of 22 g component b while stirring , and mixed intimately . the mixture is sterilized in a closed container at 121 ° c . using dry heat . the result is a homogenous , paste - like mixture that is easily extruded through a cannula , conforms to a defect due to plastic flow , and that air easily hardens superficially , it being possible to accelerate this process using a gentle airflow from a compressed air pistol . even after 4 hours of centrifuging , no solid or liquid settles out . an extruded paste adheres well to surrounding tissue . an extruded and superficially dried body retains its shape in the simulated body environment ( moist chamber under physiological conditions ). sterile filtration is performed in 1 . 9 g component h . then 5 g component b are added slowly while stirring . the result is a paste that can flow plastically and that is easily extruded through a syringe . the material can easily flow plastically into the intermediate spaces of a defect . even after multiple hours of centrifuging , the tcp particles remain in suspension . the material is very suitable for a bone construction material , especially for complicated defects . 2 . 0 g component i are mixed with component c to make 10 . 0 g and after homogenization are sterile filtered . after 25 g component b are added the material is mixed intensively . the result is a paste that has good plastic flow and that is easily extruded through a syringe . the material has plastic flow and therefore is ideal for filling intermediate spaces in a defect and adheres to the surrounding tissue . even after multiple hours of centrifuging the tcp particles remain in suspension . the material is very suitable for bone construction material , especially for complex defects . 0 . 2 g component i are mixed with component h to make 10 . 0 g and after homogenization are sterile filtered . after 25 g component b are added the material is mixed intensively . the result is a non - extrudable , kneadable paste that is slightly elastic and is shapeable like wax . because of its shapability , the material can be matched well to different defects and remains in the defect temporarily as a placeholder . even after multiple hours of centrifuging the tcp particles remain in suspension . the material is very suitable for bone construction material , especially for complex defects . 3 . 0 g component k are mixed with component c to make 10 . 0 g . once component k has dissolved , the material is sterile filtered and added to 26 . 3 g component b . the result is a paste that extrudes well through a syringe and that is shapeable and does not drip from the cannula . because of its shapability , the material can be matched well to different defects and remains temporarily in the defect as a placeholder . even after multiple hours of centrifuging the tcp particles remain in suspension . the material is very suitable for bone construction material , especially for minimally - invasive applications . 300 g component d are mixed with component c to make 10 . 0 g . after a 1 - hour swelling time , 25 g component b having the mean grain sizes according to the table below are added and mixed intimately . the result is a paste - like mass , the extrudability of which is tested using different cannula diameters . extrudability is documented in the table below and indicates the suitability of various particle diameters for producing an extrudable paste . + = good to moderate ( satisfactory extrudability , slight resistance can be detected from canting of individual particles ) − = moderate ( adequate extrudability , significant resistance due to canting of particles ) the advantage of the inventive composition is that a phase and sedimentation - stable , plastically deformable implant material with intrinsic pore formation is provided that can be created such that it does not migrate much and is inserted into a bone defect by injection or alternatively can be applied to / fitted in a bone defect as a kneadable mass . in the inventive composition , the liquid or paste components that are soluble in water stabilize the biologically active polygonally broken rounded particles that are difficult to dissolve in water and that are in the size range of 0 . 1 - 150 μm having pores in the size range of 0 . 01 - 50 μm such that they do not convert to other phases or substances due to a dissolution / recrystallization process . moreover , with the inventive application of the paste - like implant material / the inventive composition , the water - soluble organic binder substance with the particulate biologically active substance forms a porous structural accumulation that is dimensionally stable when applied , the easily soluble components being exchanged for fibrous tissue in the defect environment so that the porous particle packing remains dimensionally stable and is slowly integrated , vascularized , and resorbed . all of the features described in the following claims and in the drawings can be essential to the invention , both individually and in any combination with one another .

the invention relates to a phase and sedimentation - stable , plastically deformable preparation with intrinsic pore formation that can be used for instance for filling bone defects and for augmentation , and also relates to a method for producing same . the object of the invention , to provide a phase and sedimentation - stable , plastically deformable implant material with intrinsic pore formation , that is not complex to create , and that can be inserted into a bone defect by injection or alternatively can be applied to / fitted in a bone defect as a kneadable mass , is attained in that a plastically deformable preparation for bone construction / replacement based on ceramic suspension includes a particulate , porous component that is not easily soluble in water and at least one liquid , wax - like , or gel component that is dissolved in water or alcohol , the component that is not easily soluble in water being 60 - 98 % by weight and the composition dissolved in water or alcohol being 1 - 30 % by weight , the particles of the particulate component having a diameter in a size range that is essentially greater than a phagocytabler particle and smaller than the passage diameter of standard injection cannulas and the portion of the dissolved component in the water or alcohol being 0 . 1 - 3 % by weight .

referring to the figures where like numerals indicate the same element throughout the views , fig1 illustrates a stent - graft 10 according to the present invention . the stent - graft 10 includes a graft portion 100 and a stent portion 20 with flexible elongated rail elements 50 . the stent portion 20 provides support to the graft portion 100 when the stent - graft 10 is deployed and located in an expanded condition within a portion of a mammalian body such as a vascular lumen . the stent portion 20 includes a plurality of spaced , circumferentially extending support elements ( hoops ) 22 . each circumferential support element 22 is generally annular in shape as shown in fig1 . each circumferential support element 22 is made from a flexible , biocompatible material ( i . e ., from a material that is , for example , non - reactive and / or non - irritating ). in one embodiment , the stent portion 20 can be formed from a tube of biocompatible material . for example , the stent portion 20 can be formed by laser cutting the stent portion 20 and its support elements 22 , etc . from the tube . in another embodiment , each circumferential support element 22 is made from medical - grade metal wire formed as a closed loop ( i . e ., as an annular hoop ) in a known manner , including , for example , micro - welding two ends of a wire segment together . stainless steel , metal alloys , shape - memory alloys , super elastic alloys and polymeric materials used in conventional stents are representative examples of materials from which circumferential stent portion 20 and its support elements 22 can be formed . the alloys can include niti ( nitinol ). the polymers for circumferential support elements 22 may , for example , be bioabsorbable polymers so that the stent can be absorbed into the body instead of being removed . in a first embodiment , illustrated in fig1 and 2 , each circumferential support element 22 has a sinusoidal or otherwise undulating form , such as a wave shape . as shown in fig1 and 2 , the undulating form of the support elements 22 includes peaks 12 and troughs 13 ( space behind the peaks ). the troughs 13 include the open spaces between adjacent substantially linear struts 14 that are connected to a curved member 16 that forms the respective peak 12 . each peak 12 points in a direction that is opposite that of the immediately preceding or following , circumferentially positioned peak 12 . the same is true of the troughs 13 . each trough 13 points in a direction that is opposite the immediately preceding or following , circumferentially positioned trough . in the embodiment illustrated in fig1 and 2 , the peaks 12 all face in the one direction , toward a first end 54 of the stent 20 . similarly , the troughs 13 all face in one direction , toward a second end 56 of the stent 20 , which is opposite the first end . each circumferential support element 22 is connected to a longitudinally adjacent circumferential support element 22 by a respective bridge element 24 ( fig1 and 2 ). as shown , the bridge elements 24 connect peaks of adjacent and circumferentially out - of - phase peaks 12 of adjacent support elements 22 . as a result , adjacent support elements 22 can be rigidly spaced from each other at the area where they are joined by the bridge element 24 . in the embodiment shown in fig1 and 2 , only a limited number of bridge elements 24 are provided between respective adjacent support elements 22 . for example , adjacent support elements 22 may be connected to each other by between about one and three bridge elements 24 . in an embodiment , only one bridge element 24 extends between adjacent support elements 22 . if too many bridge elements 24 are provided between adjacent support elements , the coupling between the support elements 22 becomes similar to providing a rigid coupling between support elements , such that the desired longitudinal flexibility according to the present invention is lost . by providing only a limited number of bridge elements 24 ( including , without limitation , one bridge element 24 ), the resultant assembly can still provide a good approximation of using completely independent circumferential support elements 22 . furthermore , the peripheral location at which bridge element ( s ) 24 are provided between respective adjacent support elements 22 has an effect on longitudinal flexibility . for example , if two bridge elements are provided between a respective pair of adjacent support elements 22 at diametrically opposite sides of the support elements 22 , then , generally , the longitudinal flexibility there between is at a maximum at diametrically opposite sides of the support elements 22 located at about 90 degrees from the bridge elements 24 , and decreases along the circumference of the support elements 22 in a direction approaching the respective bridge elements 24 . for the foregoing reasons , it may be useful or otherwise beneficial to provide , for example , one bridge element 24 between adjacent support elements 22 , as illustrated in fig1 . furthermore , it may be additionally useful to offset each bridge element 24 from a longitudinally adjacent bridge element 24 in a circumferential direction , as is also illustrated in fig1 . the circumferential offset can be staggered by one set of peaks 12 along the length of the stent portion 20 between adjacent support elements 22 . alternatively , the bridge elements 24 can be circumferentially offset by up to 180 degrees for adjacent pairs of support elements 22 . the above - discussed circumferential offset embodiments provide the structural integrity benefits of using a bridge element 24 , but distribute the resultant restriction in longitudinal flexibility so that no one transverse direction of stent deflection is overly restricted . in an alternative embodiment illustrated in fig9 - 15 , the circumferential support elements 22 are formed by a plurality of connected , substantially diamond shaped support members 30 . each diamond shaped support member 30 has a first circumferential peak 32 and a second circumferential peak 33 that point in opposite circumferential directions . each support member 30 also includes a first longitudinal peak 34 and a second longitudinal peak 35 that point toward different ends of the stent portion 20 . circumferentially successive diamond shaped support members 30 are connected to each other at a junction 36 that is formed as part of the support element 22 during the pressing or molding of the support elements 22 . alternatively , the junctions 36 can be applied using conventional techniques such as welding , hooks or friction fitting . as shown in fig1 and 2 , the support elements 22 are freely mounted on flexible , elongated rail elements 50 ( hereinafter “ rails ”) such that the support elements 22 can move along the rails 50 . the rails 50 extend along the length of the stent - graft 10 between the outermost peaks 12 of terminal support elements 22 at a first end 54 and the innermost peaks 12 of the terminal support element 22 at a second end 56 . as illustrated , the terminal support elements 22 can extend beyond the terminal ends of the graft - portion 100 . rails 50 are desirably sufficiently flexible to accommodate bends , curves , etc . in a blood vessel . in one embodiment , the rails 50 are free of longitudinal expansion . also , the rails 50 may be made from , for example and without limitation the following biocompatible materials : metals , metallic alloys including those discussed above , glass or acrylic , and polymers including bioabsorbable polymers . the rails 50 can have any form . for example , the rails 50 can be solid cylindrical members , such as wires or extrusions with a circular , elliptical or other known cross sections . alternatively , the rails 50 can be ribbons or spring wires . in contrast to bridge elements 24 which are generally the same thickness and the circumferential support element 22 that they join and thus relatively inflexible , the thickness of the rails 50 can be designed to provide a desired degree of flexibility to a given stent - graft 10 . each rail 50 can have a thickness ( diameter ) of about 0 . 001 inch to about 0 . 010 inch . in an embodiment , each rail 50 has a thickness of about 0 . 0011 inch to about 0 . 005 inch . in another embodiment , each rail 50 has a thickness of about 0 . 005 inch . the rails 50 can be passed or “ snaked ” through the circumferential support elements 22 as discussed in copending u . s . patent application ser . no . 10 / 100 , 986 , which has been incorporated by reference . additionally , the rails 50 can be passed through the stent portion 20 and the graft portion 100 as discussed below . at least some of rails 50 may include end structures for preventing the circumferential support elements 22 from unintentionally passing beyond the ends 54 , 56 of the rails 50 . the end structures may have several forms as illustrated in copending u . s . patent application ser . no . 10 / 100 , 986 , which has been incorporated by reference . in an example , the end structures may be mechanical protrusions or grasp structures by which the endmost circumferential support elements 22 are fixed in place relative to the ends 54 , 56 of rails 50 . in yet another embodiment , the structures may also be a weld ( made by , for example , a laser ) for bonding a portion of an endmost circumferential support element 22 to ends 54 , 56 of rails 50 . as illustrated in fig1 , the stent portion 20 can include eight rails 50 that extend between the ends 54 , 56 . however , it is also contemplated that any number of rails 50 up to the number of peaks 12 along the circumference of the support element 22 could be used . for example , if the support elements 22 include three sets of peaks 12 , then three rails 50 could be used . if the support elements included fourteen sets of peaks 12 , then up to fourteen rails 50 could be used . in between the support elements 22 at the terminal ends 54 , 56 , the support elements 22 that are connected to each other by the bridge elements 24 are free to move along the rail ( s ) 50 . these remaining support elements 22 slide along the rail ( s ) 50 so that the stent 50 can conform to the shape of the blood vessel . it is also contemplated that the terminal support elements 22 can move along the rails 50 . in the embodiment illustrated in fig1 , the circumferential support elements 22 include apertures 17 in the curved members 16 through which the rails 50 extend . apertures 17 extend through the peaks 12 in a direction that is substantially parallel to the length of the stent portion 20 . these apertures 17 retain and orient the supporting rail ( s ) 50 in a direction parallel to the length of the stent - graft 10 . also , in an embodiment , the rails 50 are completely contained within the walls ( within the outer surface ) of the stent - graft 10 so that they do not protrude beyond the outer surface of the stent - graft 10 . the struts 14 of the stent portion 20 can have substantially any radial thickness that provides them with the needed strength to support the graft portion i 00 and a blood vessel when deployed and expanded within the vessel . each strut 14 has a substantially low profile that will not damage the vessel as it is deployed . in one example , the struts 14 can have a radial thickness of between about 0 . 0001 inch and about 0 . 020 inch . in an embodiment , the radial thickness is about 0 . 002 inch to about 0 . 008 inch . in another embodiment , the struts 14 have a radial thickness of between about 0 . 004 inch and about 0 . 005 inch . these thicknesses provide the stent - graft 10 with the needed structural and expansion properties to support the graft 100 , to support the vessel in which it is deployed and the longitudinal flexibility to conform to the natural elongated shape of the vessel . in an embodiment , the areas of the curved members 16 are formed to have the same radial thickness as that of the struts 14 in order to accommodate the apparatus 17 and the received rail ( s ) 50 . in another embodiment , the areas of the curved members 16 are formed with a greater radial thickness than the struts 14 in order to accommodate the apertures 17 . for example , the radial thickness of the curved members 16 can be between about 0 . 001 inch and about 0 . 006 inch greater than that of the struts 14 . the apertures 17 can have a diameter of about 0 . 005 inch for receiving the rails 50 . between the rails 50 where expansion occurs , the thickness could be about 0 . 004 inch . a stent portion 20 having 0 . 002 inch thick strut 14 walls could have a curved member 16 with a radial thickness of about 0 . 009 inch where the rails 50 are passed . in the embodiments illustrated in fig9 - 15 and 17 , the rails 50 extend through apertures 39 located at the first and second longitudinal peaks 34 , 35 of the support elements 22 . in a first embodiment , the areas of the support members 30 forming longitudinal peak 34 and longitudinal peak 35 and surrounding apertures 39 can have the same radial thickness as that of longitudinal struts 37 extending between the peaks 32 - 35 . in an alternative embodiment , the areas surrounding apertures 39 can have a greater radial thickness than that of the longitudinal struts 37 . as discussed above , the radial thickness of the areas surrounding apertures 39 can be between about 0 . 001 inch and about 0 . 006 inch greater than that of the struts 37 . for example , a diamond shaped support member 30 having struts 37 with a radial thickness of about 0 . 002 inch could have a longitudinal peak 34 , 35 with a radial thickness of between about 0 . 006 inch and about 0 . 009 inch . each aperture 39 can have a diameter that is large enough to slidably receive a rail 50 . the diameter of each aperture 39 can be between about 0 . 0014 inch and about 0 . 012 inch . in an embodiment , the rail receiving area has an opening of between about 0 . 0014 inch and 0 . 006 inch . however , any diameter that slidably receives a rail 50 could also be used . in alternative embodiments illustrated in fig1 - 20 , the rails 50 are slidably received within rail receiving members 130 that extend from a surface of the support member 30 forming the support element 22 . these rail receiving members 130 slidably couple a rail 50 to the support element 22 . as illustrated , the rail receiving members 130 are located proximate the longitudinal peaks 34 , 35 of their respective support member 30 . however , the rail receiving members 130 could be located at other positions along the length of their respective support elements 22 . any of the above - discussed embodiments can include support elements 22 having the rail receiving members 130 . in a first embodiment illustrated in fig1 , the rail receiving members 130 are located proximate the longitudinal peaks 34 , 35 of the support members 30 . the receiving members 130 of this embodiment include an arm 137 with a groove 139 that receives the rail 50 . the groove 139 has a bearing surface that is sized large enough to couple the support element 22 to the rail 50 , while still permitting movement of the support element 22 along the rail 50 and relative to the graft portion 100 . in the embodiment illustrated in fig1 , each receiving member 130 can include two opposing arms 158 that are offset from each other along the length of the support member 30 . like arm 137 , each arm 158 includes a groove 159 sized to couple the support member 30 to the rail 50 while permitting sliding movement of the support member and stent portion 20 relative to the rails 50 . in either embodiment illustrated in fig1 and 19 , the arms 137 , 158 can be formed by being punched , or otherwise mechanically formed , from a portion of its support member 30 . alternatively , the arms 137 , 158 could be secured to their respective support members 30 by welding or other known connection techniques . each arm 137 , 158 can be formed to extend inwardly away from its support member 30 in the direction of the graft portion 100 . in such an embodiment , the arms 137 , 158 are not intended to contact the inner surface of the vessel into which the stent - graft 10 is deployed . alternatively , the arms 137 , 158 of the receiving members 130 can project outwardly away from the stent portion 100 and the outer surface of their support members 30 that are intended to contact the inner wall of the vessel in which the stent - graft 10 is deployed . as with the above - discussed embodiments , the grooves 139 , 159 provide rail receiving areas having openings of between about 0 . 0014 inch and 0 . 012 inch . in an embodiment , the rail receiving areas of grooves 139 , 159 has an opening of between about 0 . 0014 inch and 0 . 006 inch . as illustrated in fig2 , the rail receiving members 130 can also include a pair of opposing , cooperating arms 163 that form a groove 164 into which the rail 50 can be snap fitted . the groove 164 is sized to receive the rail 50 such that the support member 30 is coupled to the rail 50 and free to move longitudinally along the rail 50 as discussed above with respect to the other embodiments . the arms 163 can be formed as discussed above with respect to the embodiments illustrated in fig1 and 19 . additionally , the arms 163 can extend from either the inner or outer surfaces of their respective support members 30 as discussed above with respect to the embodiments illustrated in fig1 and 19 . in any of the above - discussed embodiments , the illustrated graft portion 100 is formed of a well known biocompatible materials such as woven polyester including polyester terphthalate ( pet , polyester , formerly available under the dupont trademark “ dacron ”), polytetrafluroethylene ( ptfe , teflon ) and fluorinated ethylene propylene ( fep , teflon with additives for melt processing ). other polymer fabrics could be used including polypropylene , polyurethane , including porous polyurethane , and others . in an embodiment , the biocompatible material is expanded polytetrafluroethylene ( eptfe ). methods for making eptfe are well known in art , and are also described in u . s . pat . no . 4 , 187 , 390 issued to gore on feb . 5 , 1980 , which is hereby incorporated herein by reference . the graft portion 100 can be formed of either woven or a non - woven material ( s ). the porous structure of eptfe consists of nodes interconnected by very small fibrils . the eptfe material provides a number of advantages when used as a prosthetic vascular graft . the eptfe is highly biocompatible , has excellent mechanical and handling characteristics , does not require preclotting with the patient &# 39 ; s blood , heals relatively quickly following implantation , and is thromboresistant . further , eptfe has a microporous structure that allows natural tissue ingrowth and cell endothelialization once implanted into the vascular system . this contributes to long - term healing and graft patency . the graft portion 100 can be surrounded by the rails 50 and the stent portion 20 as illustrated in fig1 - 17 . in the first embodiment , illustrated in fig1 - 8 , the stent - graft 10 includes a plurality of circumferentially extending , rail receiving coupling members 60 that are spaced from each other along the length of the graft portion 100 . the rail receiving coupling members 60 eliminate the need to suture the stent portion 20 to the graft portion 100 at locations spaced from the ends of the graft portion 100 . each coupling member 60 is sized to be circumferentially and longitudinally coextensive with a portion of the outer surface of the graft portion 100 . the coupling members 60 can extend 360 degrees around the circumference of the graft portion 100 or only partially around the circumference of the graft portion 100 . for example , each coupling member 60 may extend only about 270 or 180 degrees around the circumference of the graft portion 100 . the coupling members 60 expand with the stent portion 20 and the graft portion 100 when the stent - graft 10 is expanded within a vessel using either self - expansion or a balloon . each coupling member 60 is formed of a known material such as those discussed above relating to the graft portion 100 including ptfe , eptfe , fep , woven pet ( dacron ), pet film , or any polymer that can be bonded to the exterior of the graft portion 100 and permits the smooth and easy passage of the rails 50 through their associated passageways 62 , hereinafter referred to as “ openings 62 ”. the material for each coupling member 60 can vary depending on the material used for the graft portion 100 . as shown in fig6 and 7 , the openings 62 are formed between the inner surface of the coupling member 60 and the outer surface 104 of the graft portion 100 so that the openings 62 retain their open position before and after the rails 50 have been passed through . the openings 62 are equally or unequally spaced around the circumference of the coupling members 60 . in an embodiment , the openings 62 are axially aligned along the length of the graft portion 100 . however , in an alternative embodiment , the openings 62 of adjacent coupling members 60 can be circumferentially offset relative to each other . the number of openings 62 circumferentially spaced about the coupling member 60 will equal the number of rails used for the stent - graft 10 . for example , if the stent - graft 10 includes five rails 50 , then each longitudinally spaced coupling member 60 could include at least five openings 62 . in an embodiment , the number of coupling members 60 will be equal to the number of support elements 22 that extend around the graft portion 100 . as illustrated in fig5 , each coupling member 60 is formed of a single layer 64 of material secured to the outer surface of the graft portion 100 by ultrasonic welding , adhesive bonding , thermal fusing or other known manners . in this embodiment , the rails 50 extend between the inner surface 63 of each coupling member 60 at a respective opening 62 and the outer surface 104 of the graft portion 100 . in an alternative embodiment , the coupling member 60 includes a first circumferentially extending member secured to the outer surface 104 of the graft portion 100 and a second circumferentially extending member positioned over the first member . in this embodiment , the openings 62 are formed between the two circumferentially extending members . in any of the above embodiments relating to fig1 - 8 , the coupling members 60 are secured to the graft portion 100 and the stent portion 20 while receiving the rails 50 so that the coupling members 60 can move along and relative to the rails 50 . the coupling members 60 can be secured to the support elements 22 by welding or other known conventional securing techniques . in an alternative embodiment , the coupling members 60 can extend through slots in the support elements 22 or they can be adhesively secured in recesses formed on the inner surfaces of the support elements 22 . in the alternative embodiment illustrated in fig9 - 11 , the coupling members 60 can be positioned along the length of the stent - graft 10 and oriented so that their openings 62 are circumferentially offset from the openings 62 of longitudinally adjacent coupling member ( s ) 66 , 68 . as shown in fig9 , coupling member 66 can have openings 62 that are positioned within the openings in circumferentially spaced support members 30 so that a respective rail 50 passes through the opening 62 in the coupling member 60 at point a that is between the longitudinal peaks 34 , 35 of the support members 30 . the coupling member 60 then passes under the circumferentially adjacent rail ( s ) 50 that extends through the immediately , circumferentially adjacent support member ( s ) 30 ( see fig9 ). the openings 62 of the immediately , longitudinally adjacent coupling member 68 are circumferentially offset from those of coupling member 66 so that the rail 50 passes through the openings 62 of the adjacent coupling member 68 at point b . as a result , immediately , longitudinally adjacent coupling members 60 ( 66 , 68 ) slidably receive circumferentially spaced rails 50 at offset points . this can increase the stability of the stent - graft 10 without reducing its ability to conform to the shape of the vessel in which it is deployed . in an alternative embodiment , shown in fig1 and 11 , the longitudinally spaced coupling members 60 receive the rails 50 outside the support members 30 at point b . in this embodiment , the openings 62 of longitudinally adjacent coupling members 60 are circumferentially and longitudinally aligned . in the embodiments illustrated in fig1 - 15 , the rails 50 could extend through cauterized openings in the graft portion 100 in place of using the coupling members 60 . hence , in these alternative embodiments , immediately , circumferentially adjacent rails 50 could be extended through cauterized openings 80 in the graft portion 100 at longitudinally and / or circumferentially offset points ( a , b ) as shown in fig9 and 12 . alternatively , the adjacent rails 50 could be extended through cauterized openings 80 the graft portion 100 at circumferentially and / or longitudinally aligned locations b , as shown in fig1 . in any of the above - discussed embodiments , the graft portion 100 will move with support elements 22 as the support elements 22 move along the rails 50 . in the embodiment illustrated in fig1 and 17 , the rails 50 pass through circumferentially extending retainer coupling members 200 , hereinafter referred to as “ loops 200 ”. unlike coupling members 60 shown in fig9 , the loops 200 have interior regions 202 that pass through openings 195 in the graft portion 100 and extend along an inner surface of the graft portion 100 . the openings 195 can be welded , cauterized or otherwise closed about the loops 200 using other known techniques . in an embodiment , the loops 200 can be formed of yam that is stronger than the graft portion 100 . in an embodiment , the loops 200 are formed of a pet , 80 denier loop yam . the loops 200 can also be formed of any of the materials discussed above with respect to the graft portion 100 . the loops 200 can also be formed of a solid polymer fiber , braid , film , or the like . it is also possible to bond or otherwise secure the loops 200 to the graft portion 100 . portions of the loops 200 on the exterior of the graft portion 100 and in - between the interior regions 202 form arches 210 along the outer surface of the graft portion 100 . the arches 210 slidably receive the rails 50 so that the graft portion 100 can move along the rails 50 and relative to the support elements 22 . while rounded arches 210 are illustrated , any shaped opening that slidably receives the rails 50 can be used . for example , the opening of the arches 210 can include a rectangular , elliptical or triangular shape . the arches 210 each include an opening sized to receive the rails 50 . these opening can be between about 0 . 0014 inch and about 0 . 012 inch . in an embodiment , the arch openings can be between about 0 . 0014 inch and about 0 . 006 inch . in an embodiment , the arch openings can be about 0 . 005 inch . each arch 210 is spaced from circumferentially spaced arches 210 by a distance that is substantially equal to the circumferential spacing of the adjacent rails 50 . the adjacent arches 210 can be equally spaced from each other around the circumference of the graft portion 100 . alternatively , adjacent arches 210 can be circumferentially spaced at different intervals around the circumference of the graft portion 100 to provide different flexion capabilities to the stent graft 10 . each arch 210 can be spaced from an adjacent arch 210 by a distance of about 0 . 10 inch to about 0 . 30 inch . in one embodiment , adjacent arches 210 are spaced from each other by a distance of about 0 . 155 inch . the support elements 22 comprise the diamond shaped support members 30 shown in fig9 and 17 . however , as with the above - discussed embodiments , other known shapes may also be used . similar to the embodiments illustrated in fig9 - 15 , the support elements 22 shown in fig1 include apertures 39 and are free of a connection to the loops 200 . the support elements 22 ( fig1 ) are moveable along the rails 50 in a direction that is substantially parallel to the length of the graft portion 100 as discussed above . the movement of the support elements 22 along the length of the stent - graft 10 and relative to the rails 50 and graft portion 100 can be limited by one or both of the longitudinal peaks 34 , 35 abutting against a support element 200 . as shown in fig1 , the arches 210 of the loops 200 can act as a stop for the longitudinal movement of the support element 22 . therefore , the total distance that the support elements 22 move along the rails 50 can be controlled and limited by the spacing between the loops 200 along the length of the graft portion 100 . in one embodiment , each loop 200 can be spaced from adjacent loops 200 along the length of the graft portion 100 by the same distance as the coupling members 60 so that the support elements 22 can move a distance that permits the stent - graft 10 to conform to the shape of the vessel in which the stent - graft 10 is deployed . the spacing between adjacent loops 200 ( and 60 ) can be less than the distance that each support element 22 extends in a direction parallel to the length of the stent - graft 10 . unlike the other embodiments ( for example the embodiment illustrated in fig1 ), each support elements 22 illustrated in fig1 is free of a connection to a longitudinally adjacent support element 22 by a bridging element . as a result , the support elements 22 , illustrated in fig1 , can move independently relative to each other along the length of the graft portion 100 . also , like the embodiments discussed above , the rails 50 can include a single , continuous member with multiple turns ( fig1 ), a plurality of separate members with at least one turn that are circumferentially spaced from adjacent members around the graft portion 100 , or separate , individual members that are free of turns and that are free of a direct , secured attachment to an adjacent rail 50 . as used herein , the term “ rail ” includes each of these arrangements . in another alternative embodiment , the graft portion 100 can include integral , spaced areas that receive the rails 50 formed of the material used to form the graft portion 100 . these spaced areas have an increased thickness with respect to the remainder of the graft portion 100 . the present invention also includes introducing an agent , including those set forth in u . s . patent application ser . no . 60 / 426 , 366 , which is hereby incorporated by reference , into a body using the above - discussed stent - graft 10 . in a preferred embodiment , the agent ( s ) is carried by one or more of the rails 50 or the graft portion 100 and released within the body over a predetermined period of time . for example , these stents can deliver one or more known agents , including therapeutic and pharmaceutical drugs , at a site of contact with a portion of the vasculature system or when released from a carrier as is known . these agents can include any known therapeutic drugs , antiplatelet agents , anticoagulant agents , antimicrobial agents , antimetabolic agents and proteins . these agents can also include any of those disclosed in u . s . pat . no . 6 , 153 , 252 to hossainy et al . and u . s . pat . no . 5 , 833 , 651 to donovan et al ., both of which are hereby incorporated by reference in their entirety . local delivery of these agents is advantageous in that their effective local concentration is much higher when delivered by the stent than that normally achieved by systemic administration . the rails 50 , which have a relatively low elastic modulus ( i . e . low force to elastic deformation ) in their transverse direction , may carry one or more of the above - referenced agents for applying to a vessel as the vessel moves into contact with the agent carrying rail ( s ) 50 after deployment of the stent - graft 10 within the vessel . these agents can be applied using a known method such as dipping , spraying , impregnation or any other technique described in the above - mentioned patents and patent applications that have been incorporated by reference . applying the agents to the rails 50 avoids the stresses at focal areas as seen in the struts of traditional stents . in this manner drug coatings applied to the stent rails 50 may be used with support elements formed of materials that are otherwise unsuitable for coating . it is contemplated that the various elements of the present invention can be combined with each other to provide the desired flexibility . for example , the rails 50 can be formed of one or more radiopaque materials . additionally , the support element designs can be altered and various support element designs that permit the passage of the rails could be used . similarly , the number , shape , composition and spacing of the rail elements can be altered to provide the stent with different properties . additionally , the device can have varying numbers and placement of the bridge elements . the properties of any individual stent would be a function of the design , composition and spacing of the support elements , rails and bridge elements . thus , while there have been shown and described and pointed out fundamental novel features of the present invention as applied to preferred embodiments thereof , it will be understood that various omissions and substitutions and changes in the form and details of the devices illustrated , and in their operation , and in the method illustrated and described , may be made by those skilled in the art without departing from the spirit of the invention as broadly disclosed herein .

a stent - graft with increased longitudinal flexibility that is deployed within a body lumen for supporting the lumen and repairing luminal aneurysms . in a preferred embodiment , the stent - graft is located and expanded within a blood vessel to repair aortic aneurysms . the stent - graft is comprised of an expandable stent portion , an expandable graft portion and at least one elongated rail . the stent portion and graft portion are moveable between the terminal ends of the rail and relative to the rails so that it can conform to the shape of a vessel in which it is deployed . the stent - graft provides increased longitudinal flexibility within a vessel . also , the stent - graft of the present invention does not kink after expansion , and thus , eliminates the potential for the graft portion occluding the blood flow lumen of the vessel in which it is deployed . moreover , the wear on the graft is reduced and its longevity increased .

referring to fig1 - 3 , molded plastic shoe insert 10 includes thin body 11 , three ( 3 ) dumbbell - shaped rib portions 12 , 13 , and 14 , and circular raised portion 16 . formed as part of the rib portions 12 , 13 , 14 are threaded sockets 17 , 18 , 19 , 20 , 21 , 22 , and 23 . rib portions 12 , 13 , and 14 have generally circular reinforcing elements 17a , 18a , 19a , 20a , 21a , and 22a positioned around the respective sockets . rib portions 12 , 13 , and 14 also include rib crosspieces 12r , 13r , and 14r . also shown is shoe spike - carrying unit 26 including metal spike pin 27 with metal base 28 and plastic cap 29 . plastic cap 29 has two ( 2 ) wrench - receiving holes 30a , 30b for receiving a tool to screw spike 26 in to install for use and screw it out for removal and replacement . turning in particular to fig1 and 2 , the dumbbell rib construction of sole plate 10 permits plate body 11 to be made thinner than a sole without such construction . the present construction provides for a lighter weight and more flexible sole plate . ribs 12 , 13 and 14 also provide spaced - apart support surfaces for a rubber , leather ( or other material ) outer shoe sole 36 ( fig2 ). the spacing between outer sole socket receiving holes 37 permits relative movement between sole 36 and insert 10 . such relative movement is accomplished with less required forces as compared with a flat non - rib insert plate surface thus providing more flexibility to sole construction . turning to fig4 spike 26 is shown in its initial step of being screwed into socket 23 . socket 23 which extends upwardly from circular raised portion 16 includes spaced - apart upstanding socket side projections 41 having slanted upper cam surfaces 41a and vertical non - cam surfaces 41b . spike element 26 carries circular skirt 32 with flexible frangible lock tongues 33 depending in spaced manner from skirt 32 . the original shape of lock tongue 33 includes cam surface 33a and vertical non - cam surface 33b . when spike 26 is initially being screwed into socket 23 skirt 32 and the top rim 23a of socket 23 are spaced apart . in fig4 as spike 26 is screwed on in the direction of the arrow , lock tongues 33 just clear upstanding side rib 41 . in fig5 after spike 26 has been screwed on further and spike skirt 32 is just above the upper rim 23a of socket 23 , lock tongues 33 are forced to be deformed by camming action of upstanding projection cam surfaces 41a . with further turning the lock tongues 33 pass the ribs 41 and restore themselves to some extent to their original shape ( fig6 ). spike 26 is turned until skirt 32 engages socket upper rim 23a or until the spike 26 is otherwise tightly held in place . each expendable tongue 33 will in this installation process pass against , be deformed by , and pass by a number of socket projections 41 a distance x &# 39 ; from rim 23a . the interference between projections 41 and tongues 33 as spike 26 is urged to be turned holds spike 26 in place during shoe use . when replacement of spike 26 is desired , a wrench is used to turn the spike in the opposite direction ( arrow of fig7 ). since the tongue vertical side 33b is presented to the vertical non - cam side 41b of the projection 41 , a portion of tongue 33 will shear off leaving tongue stump 33s which stump 33s can clear the projections 41 and permit removal of the spike 26 . it is also contemplated that the depending tongues may be made of selected materials and so shaped such that the tongues will undergo distortion as the spike is installed and when the spike is removed will undergo additional distortion without shearing or breaking .

a spike shoe construction including a socket carrying a ribbed insert mounted on and spaced from an outer sole with each socket carrying a reusable lock for locking spikes installed in the socket . each spike in turn carries a lock which is non - reusable in that it is so distorted or broken when removed to render it inoperative .

for the purposes of promoting an understanding of the principles of the invention , reference will now be made to the embodiment illustrated in the drawings and specific language will be used to describe the same . it will nevertheless be understood that no limitation of the scope of the invention is thereby intended , such alterations and further modifications in the illustrated device , and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates . the preferred embodiment of the method of the present invention will now be described with reference to fig1 a - d which illustrate sequential stages in the performance of this invagination technique at the gastroesophageal junction 10 which anatomically connects the esophagus 20 to the stomach 30 . preliminarily , two operating channel / insufflation ports are percutaneously opened into the stomach 30 , one port being sized to accommodate an endoscope ( 7 mm ), and the second port being about 12 mm in diameter to accommodate a remotely operable fastening assembly . this is generally accomplished by 1 ) percutaneously introducing a guide wire into the stomach , 2 ) using an endoscopic to snare the guide wire and retract it through the mouth , 3 ) advancing a graduated dilator operating channel port assembly transorally over the guide wire under tension and partially out through the skin , and 4 ) anchoring the assembly into place . the steps of this aspect of the procedure will be more fully discussed later in this specification in relation to fig3 a - d . after the stomach has been insufflated through the insufflation lumens of the previously implanted ports , another needle is placed through the abdominal wall into the stomach and under endoscopic guidance a guide wire 5 is introduced and pulled out the mouth . invagination device 70 is then passed over guide wire 5 and into esophagus 20 . the introduction and advancement of invagination device 70 into esophagus 20 is facilitated by the flexibility of device 70 along its length and by the tapered shape of its distal end . guidance of device 70 through esophagus 20 can be further facilitated by the application of appropriate tension on guide wire 5 while device 70 is being advanced . a 5 mm laparoscope 40 is introduced through the 7 mm port and invagination device 70 is positioned , under direct observation by laparoscope 40 , at the anatomical gastroesophageal junction 10 . the location of invagination device 70 in relation to gastroesophageal junction 10 is facilitated by the affixation of a marking 71 at a predetermined distance ( 3 cm ) from the distal end of device 70 . fig1 a shows invagination device 70 having been advanced into esophagus 20 and so positioned at the gastroesophageal junction 10 . the engagement assembly of invagination device 70 is then activated to advance needles 88 out through lumens 78 to project radially and forwardly from invagination device 70 and into engagement with esophagus 20 . the mechanism for projecting needles 88 into engagement with esophagus 20 will be described more specifically later in this specification in relation to fig2 a - b . fig1 b shows invagination device 70 with its engagement assembly having been so activated to extend needles 88 into engagement with esophagus 20 in the vicinity of the gastroesophageal junction . it is to be noted that fig1 b conceptually shows this engagement and is not drawn to scale . in practice , the engagement of needles 88 with esophagus 20 should be made as close to esophageal junction 10 as practicable . most preferably , engagement should be made directly at the gastroesophageal junction 10 in order to prevent long term exposure of the esophogeal inner lining to the stomach secretions . once so engaged , invagination device 70 is pushed forward toward stomach 30 , which movement forces esophagus 20 toward stomach 30 to invaginate the gastroesophageal junction 10 and involute the surrounding fundic wall 25 . a 10 mm remotely operable stapling device 50 is then advanced through the previously opened 12 mm operating channel / insufflation port , and operated to staple fundic wall 25 to the invaginated gastroesophageal junction 10 . grooves 72 at the tapered distal portion of introducer guide 75 serve to facilitate the stapling action of stapler 50 by providing a stable backing therefor . fig1 c shows engaged invagination device 70 having been advanced toward stomach 30 to so invaginate gastroesophageal junction 10 , and fig1 c further illustrates fundic wall 25 being fastened to the invaginated gastroesophageal junction 10 by stapler 50 . lastly , stapler 50 is removed , needles 88 are retracted back into their respective lumens 78 , and invagination device 70 and guide wire 5 are removed . laparoscope 40 is also removed . operating channel / insufflation ports then sealed and are kept in place for a period of time to maintain fixation of the newly formed valve . fig1 d shows the newly formed valve between esophagus 20 and stomach 30 , with invagination device 70 and other instrumentation having been removed , and with staples 55 holding fundic wall 25 to the gastroesophageal junction 10 . it is believed that two or three rows of staples 55 should be sufficient to hold the formed fold in place and maintain the functionality of the so formed valve . invagination device 70 will now be discussed in specific detail in relation to fig2 a - b , which show partially fragmented and segmented side elevational views of this device which is used in the above described method . fig2 a shows the device with its engagement assembly deactivated , the engagement needles being retained within their respective lumens . fig2 b shows the device with its engagement assembly activated to project the needles from their respective lumens for engagement with the esophageal wall . invagination device 70 includes introducer guide 75 which has an outside diameter approximating that of the esophagus . introducer guide 75 is made with material that is biocompatible and flexible ( low density polyethylene is particularly suited for this purpose ), and is tapered at its distal end to facilitate insertion and advancement into the esophagus . introducer guide 75 defines a central lumen 76 , extending the length thereof , through which a guide wire is receivable . the central lumen 76 of introducer guide 75 also accommodates needle projector 80 . extending from central lumen 76 in introducer guide 75 are ten needle - receiving lumens 78 , which are equidistantly spaced about the circumference of introducer guide 75 and extend to the outer diametric surface thereof . at the proximal end of introducer guide 75 is a male threaded connector 77 . invagination device includes an engagement assembly which can be activated to project needles 88 out of their respective needle - receiving lumens 78 and into engagement with the esophagus . this engagement assembly for projecting needles 88 includes needle projector 80 , advancing member 90 , and connector 77 . needle projector 80 has a metal hub 81 at its distal end to which needles 88 are connected . at its proximal end , needle projector 80 has a second metal hub 82 which is seated within recess 93 of advancing member 90 . extending along its length , the shaft 83 of needle projector 80 has a metallic interior core 84 and a plastic outer tubing 85 . metallic interior core 84 defines a central lumen 86 therein through which a guide wire is receivable . metallic interior core 84 is made of stainless steel and provides axial strength and rigidity to shaft 83 , while plastic outer tubing 85 , which is constructed from low to medium density polyethylene , enhances the flexibility of shaft 83 and provides for laterally stability within the central lumen 76 of introducer guide 75 . together , hubs 81 and 82 and shaft 83 form a &# 34 ; push rod &# 34 ; which , when advanced within central lumen 76 of guide 75 , projects needles 88 out of their respective needle - receiving lumens 78 and into engagement with the esophagus . the advancement of projector 80 within central lumen 76 is accomplished by the operation of advancing member 90 . advancing member 90 has , at its distal end , a female threaded portion 91 which threadedly engages the male threaded portion 79 of connector 77 and , at its proximal end , retaining portion 92 which retains hub 82 within recess 93 . central lumen 96 through advancing member 90 receives shaft 83 and proximal hub 82 of needle projector 80 , and continues through the proximal end of advancing member 90 to allow a guide wire to be passed through the entire length of invagination device 70 . by threading advancing member 90 over connector 77 of introducer guide 75 , needle projector 80 is advanced within introducer guide 75 to project needles 88 out through needle - receiving lumens 78 , extending needles 88 radially and forwardly about the circumference of device 70 for engagement with the esophagus . when advancing member 90 is fully threadedly engaged over connector 77 , needles 88 are fully extended from introducer guide 75 . by the use of the threaded engagement for advancement , needles 88 are projected smoothly and evenly to provide for a steady and sure engagement with the esophageal wall . needles 88 are guided by needle - receiving lumens 88 out at an angle of about 30 ° to the longitudinal axis of device 70 . ten needles , equidistantly spaced about the circumference of device 70 , 36 ° apart , at this projection angle of 30 °, have been found to be sufficient to stably engage and advance the esophageal wall . an operating channel / insufflation port assembly useful for the introduction of laparoscope 40 and remotely operable stapling assembly 50 will now be discussed in specific detail in relation to fig3 a - d . operating channel / insufflation port assembly 100 includes guide wire 105 , operating channel / insufflation port 110 , skin flange 120 , attachable balloon inflator nozzle 130 , attachable insufflation valve 140 , sealing member 150 , and seal cap 160 . procedurally , guide wire 105 is first percutaneously introduced into the stomach , and an endoscope is introduced transorally into the stomach and used to snare guide wire 105 and retract it through the mouth . operating channel / insufflation port 110 is then transorally advanced over guide wire 105 under tension and partially out through the skin . after removing guide wire 105 , skin flange 120 is placed over the tubular portion of port 110 and advanced toward the skin . attachable balloon inflator nozzle 130 is then attached to inflation port 112 and co 2 is injected therethrough and into balloon 118 , through inflation lumen 113 which is connected thereto , thus inflating balloon 118 in place within the stomach . skin flange 120 is then adjusted against the skin to secure port 110 into place . the tapered distal portion 111 of operating channel / insufflation port 110 is then cut off at mark 111 &# 39 ;, thereby opening access into the stomach through operating channel 114 . when connected to insufflation port 115 , insufflation valve 140 is operated to provide a supply of co 2 into the stomach through insufflation lumen 116 and out insufflation opening 117 . by turning cock 142 , the supply of co 2 into the stomach may be variably controlled and adjusted as needed . operating channel / insufflation port 110 is preferably made of flexible material which matches the elasticity of the tissue surrounding it when implanted . silicone is believed to be particularly suited for this purpose . to provide additional strength and stability during implantation , tapered portion 111 of port 110 has a lumen 104 of only enough size to receive wire guide 105 . as previously discussed , tapered portion 111 is severed after implantation to expose central operating channel 114 , thereby providing access into the stomach for remotely operable instrumentation which is to be used in conjunction with the operation to be performed . skin flange 120 is also made of silicone , and includes of a disc shaped flange 121 and a tubular handle portion 122 . slot 123 facilitates the advancing of flange 120 along port 110 . seal cap 150 seals the end of port 110 , attaching thereabout to form a sealing fit between detent 151 of seal cap 150 and indentation 119 of port 110 . fig3 b - d illustrate enlarged cross - sectional views of portions of the assembly at various stages of operation . fig3 b shows an implanted operating channel / insufflation port 110 , balloon inflation nozzle 130 , and insufflation valve 140 , with nozzle 130 and valve 140 detached from their respective connection ports . in fig3 b it can be seen that tapered portion 111 has a central lumen 104 sized only to receive guide wire 105 , thus providing additional strength and stability during implantation , while tubular portion 110a defines an enlarged operating channel 114 , sized to receive remotely operable instrumentation ( 12 mm , for example , for a fastening assembly , and 7 mm , for a laparoscope ). within the walls of said tubular portion 110a are defined a balloon inflation port 112 and balloon inflation lumen 113 which connects port 112 to the interior of balloon 118 . at the entrance of port 112 is seal 112a . also defined within the walls of tubular portion 110a is an insufflation lumen 116 , connecting exterior insufflation port 115 to interior insufflation port 117 . at the entrance of port 115 is seal 115a . also shown in fig3 b - d are partial cross - sectional views of balloon inflation nozzle 130 and insufflation valve 140 , which are connectable to ports 112 and 115 respectively . when placed into port 115 , as shown in fig3 c , seal 115a is opened , allowing co 2 to be injected into the stomach through valve passageway 141 and insufflation lumen 116 . when valve 140 has been removed from port 115 , as shown in fig3 d , seal 115a recloses . balloon inflation nozzle 130 operates in the same manner in relation to port 112 to inflate balloon 118 through passageway 131 and inflation lumen 113 . in fig3 c , tapered portion 111 of port 110 has been severed , and fastening assembly 50 has been inserted through central operating channel 114 . also in fig3 c , insufflation valve 140 has been attached to insufflation port 115 for insufflation of the stomach through insufflation lumen 116 . sealing member 150 has been placed about port 110 and positioned in sealing engagement with fastening assembly 50 , thus preventing gas leakage through operating channel 114 during operation / insufflation . detent 151 of sealing member 150 fits in indentation 119 of port 110 in sealing engagement , while interior flange 152 forms a sealing engagement with fastening assembly 50 to effectly prevent leakage . in fig3 d , fastening assembly 50 has been removed , insufflation valve 140 has been detached , and seal cap 160 has been placed about port 110 , sealing access to and from the stomach . so sealed , port 110 may be kept implanted for a period of time after completion of the above described operation in order to maintain fixation of the stomach to the peritoneum and also provides a drainage port for the stomach . in the same manner as fig1 a - d , fig4 a - d illustrate sequential stages in the performance of an invagination technique according to the present invention in conjunction with an alternatively configured invagination device . except as otherwise noted , all aspects of fig4 a - d are the same as fig1 a - d , with the primed notations of fig4 a - d corresponding the same relative unprimed notations of fig1 a - d . therefore , to avoid redundancy the description of these additional drawings will not be repeated except to the extent of the following differences . specifically , in fig4 a - b it is to be noted that groove 72 &# 39 ; extends longitudinally from the beginning of the taper in guide 70 &# 39 ; proximally past lumens 78 &# 39 ; and that lumens 78 &# 39 ; have been reconfigured to provide room for groove 72 &# 39 ; to extend therepast . this revised configuration facilitates placement of the anvil jaw of fastening assembly 50 &# 39 ; within the esophgeal lumen and allows fastening assembly 50 &# 39 ; to be positioned past needles 88 &# 39 ; during stapling , as shown in fig4 c , thus allowing the needle engagement to be made directly at the gastroesophogeal junction 10 . engagement directly at the gastroesophageal junction 10 prevents long term exposure of the esophogeal inner lining to the stomach secretions . fig5 a shows an enlarged end view of invagination guide 70 &# 39 ;, illustrating the circumferential spacing of lumens 78 &# 39 ; and grooves 72 &# 39 ;. fig5 b is an enlarged fragmented side elevational view of invagination guide 70 &# 39 ; showing groove 72 &# 39 ; extending past lumens 78 &# 39 ;. while the invention has been illustrated and described in detail in the drawings and foregoing description , the same is to be considered as illustrative and not restrictive in character , it being understood that only the preferred embodiment has been shown and described and that all changes and modifications that come within the spirit of the invention are desired to be protected .

a technique for invaginating the esophagus at the gastroesophageal junction is disclosed , as well as an invagination device useful in the described technique . the invagination device includes an introducer guide which has an outside diameter approximating that of the esophagus and a plurality of needle - receiving lumens extending to its outer diametric surface . the invagination device also has an engagement assembly which includes 10 needles , each having a retracted position in which they lie within the needle - receiving lumens of the introducer guide , and an extended position in which they extend out of the lumens and project radially from the guide for engagement with the esophagus at the gastroesophageal junction . the invagination device is introduced transorally into the esophagus , its engagement assembly is activated to place the needles in their extended position in engagement with the esophagus , and the engaged invagination device is advanced the toward the stomach to fold the attached esophagus beyond the gastroesophageal junction . a remotely operable fastening assembly , which has been introduced into the stomach through an operating channel / insufflation port , is then operated to fasten the invaginated gastroesophageal junction to the surrounding involuted fundic wall . finally , the needles are retracted back into their needle - receiving lumens and the invagination device and the fastening assembly are removed from the body of the patient . this minimally - invasive technique accomplishes the formation of a new valve between the esophagus and stomach , thus preventing acid reflux .

referring to fig1 , a block diagram of an ablation system according to an exemplary embodiment is shown . as shown in fig1 , ablation system 10 comprises ablation controller circuitry 3 , an ablation energy generator 5 in electronic communication with ablation controller circuitry 3 , and an ablation electrode 7 in electronic communication with ablation energy generator 5 . according to an exemplary embodiment , ablation energy generator 5 provides energy and directs the energy to ablation electrode 7 to ablate tissue . according to various exemplary embodiments , any suitable generator may be used such as dc energy pulse from a defibrillator , rf transmitter , microwave transmitter , laser light source , etc . according to various exemplary embodiments , any suitable type of energy may be used for ablating tissue . according to an exemplary embodiment , ablation system 10 comprises a light source 41 and a light receptor 33 in communication with controller circuitry 3 . light source 41 , which directs ( nir ) energy towards the heart tissue and the light receptors 33 , is operable to detect the incidence of light reflected from tissue . ablation system 10 may also include cardiac sensing circuitry 11 . according to an exemplary embodiment , cardiac sensing circuitry 11 may , for example , detect atrial and / or ventricular activity and may provide an ecg waveform signal representative of cardiac activity . according to various exemplary embodiments , the cardiac sensing circuitry may include any circuitry known in the art for detecting cardiac activity . as shown in fig2 , ablation electrode 13 attaches to and / or forms a part of a catheter 15 or another medical device configured to be positioned within the heart . according to various exemplary embodiments , the ablation electrode may comprise any suitable electrode . according to an exemplary embodiment , ablation catheter 15 as shown in fig2 comprises a catheter body 17 having a proximal end 19 , a distal end 21 and at least one lumen 23 extending substantially through the catheter body 17 , a plurality of openings 25 in the surface of the catheter 15 extending through the outside surface of the catheter into lumen 23 , and a system for introduction of a conductive media into lumen 23 , whereby the conductive media passes through lumen 23 and is expelled out one or more openings 25 in catheter 15 to contact tissue 27 to be ablated . according to various exemplary embodiments , any suitable system for controlling the flow of the conductive media through the lumen to create a consistent flow of conductive media out through the one or more openings in the catheter may be used . according to an exemplary embodiment , catheter body 15 comprises an elongated catheter made of materials suitable for use in humans , such as nonconductive polymers . according to a particularly preferred embodiment , the catheter is between 0 . 5 meters and 1 . 5 meters in length and between 1⅓ mm and 5 mm in diameter . according to a particularly preferred embodiment , the catheter contains one or more lumens sufficient to accommodate wires for one or more sensing electrodes , such as thermosensing devices . according to an exemplary embodiment , catheter body 17 comprises a system for controlling movement of the catheter 15 such as a pull wire ( not shown ). according to various exemplary embodiments , any suitable catheter may be used . referring to fig2 , ( nir ) energy is projected through a fiber optic channel 31 provided in catheter 15 ( e . g ., a cardiac catheter ) to characterize tissue at distal end 21 of catheter 15 . according to an exemplary embodiment , catheter 15 is configured to use ( nir ) spectroscopy to provide real - time spectrograms displaying tissue properties . depending on the characteristics of the heart tissue , the tissue absorbs some of the light and reflects some of the light back toward catheter 15 . the light reflected back toward catheter 15 is received by light receiving receptors 33 attached to catheter 15 . referring to fig2 - 3 , catheter 15 comprises a light emitting fiber 35 . according to an exemplary embodiment , catheter 15 comprises a plurality of light receiving fibers 37 surrounding the light emitting fiber 35 in a bundle of fibers 39 as shown in fig3 . according to an exemplary embodiment , light emitting fiber 35 comprises a remote light source 41 as shown in fig2 . fig2 shows a display 45 of system 10 , which is illustrated separate from catheter 15 and may be located in a convenient and suitable viewing area for the physician or clinician performing an ablation procedure . according to an exemplary embodiment , spectrometer 43 is in electrical communication with light receiving . fibers 37 . light reflected from the tissue strikes a photodiode ( not shown ), which converts the light energy to data or an electrical signal so that the light may be processed by spectrometer 43 . the electrical signal is then amplified and displayed on spectrograph 43 . fig3 shows a cross - sectional view of light emitting fiber 35 and light receiving fibers 37 according to an exemplary embodiment . according to various exemplary embodiments , data collection is nearly instantaneous , the ( nir ) spectrogram has a high signal to noise ratio , good resolution , is small enough in physical size that it can be inserted into the body via a catheter , etc . according to an exemplary embodiment , the ( nir ) spectrometer has resolution & lt ; 30 m and a speed of between 5 - 10 ms per scan . the range of the spectrometer is between 1 - 2 . 5 μm . according to an exemplary embodiment , the spectrometer signal can be transmitted wirelessly outside of the body to an adjacent spectrograph or alternatively from the distal end of the catheter through fiber optic cable or other suitable wire to the spectrogram outside of the body . in the embodiment shown , the ( nir ) energy is passed through fiber optic channel 31 to distal end 21 of catheter 15 into the heart tissue . according to various exemplary embodiments , the ( nir ) energy light source may be emitted via semiconductor , tungsten - halogen , etc . as shown in fig2 , light source 41 is relayed via fiber optic cable 35 and directed towards heart tissue 27 . some light is absorbed by heart tissue 27 and some light is reflected back towards the light receiving fibers 37 . according to an exemplary embodiment , this reflected light is absorbed by light receiving fibers 37 and transmitted back to spectrometer 43 via a fiber optic connection fiber 38 , where it is directed to a single element indium gallium arsenide ( ingaas ) detector . according to an exemplary embodiment , the ingaas detector comprises a photodiode that converts the light into electrical signals , which are then amplified by a transimpedance amplifier for processing . referring back to fig1 , the signal received is then processed by controller circuitry 3 . controller circuitry 3 is operable to recognize various input signals and various sensed signals as well as being operable to filter and display the signals on a spectrograph . according to various exemplary embodiments , the controller circuitry may be operable to perform various suitable processing and analysis functions such as fourrier analysis , etc . to determine levels of ablation of the affected tissue . fig4 is a flow chart showing a process 100 for ablating and characterizing tissue according to an exemplary embodiment . in a cardiac ablation procedure , ablation is commenced . as ablation begins , a light emitting fiber directs ( nir ) energy towards the heart tissue being ablated . the ( nir ) energy is partially absorbed by the heart tissue and partially reflected towards the ablation catheter ( step 101 ). light receptors receive the reflected energy and the light is conveyed to a photodiode via a light receiving fiber where ( nir ) energy is detected ( step 103 ). the photodiode converts said light into an electrical signal , which may be amplified , filtered and / or processed such that an output can be analyzed and / or displayed on a spectrograph 43 ( step 105 ). based on the data provided by the spectrograph , a determination may be made whether the tissue is sufficiently ( e . g ., fully ) ablated ( step 107 ). once a determination has been made , the ablation power may be adjusted ( e . g ., automatically , manually , etc .) according to a desired ablation level ( step 109 ). according to an exemplary embodiment , the method of the present invention may include an algorithm for identifying ablated tissue and reducing ablation energy . according to various exemplary embodiments , the method may additionally comprise the steps of displaying an automated message to the clinician regarding the level of tissue ablation , and / or reducing power to the ablation electrode if the tissue ablation data reaches predefined criteria ( e . g ., indications of full ablation ). the construction and arrangement of the elements of the system as shown in the detailed description is illustrative only . although only a few embodiments have been described in detail in this disclosure , those skilled in the art who review this disclosure will readily appreciate that many modifications are possible ( e . g ., variations in sizes , dimensions , structures , shapes and proportions of the various elements , values of parameters , mounting arrangements , use of materials , orientations , etc .) without materially departing from the novel teachings and advantages of the subject matter recited . for example , elements shown as integrally formed may be constructed of multiple parts or elements shown as multiple parts may be integrally formed , the operation of fasteners , connectors , etc . may be reversed or otherwise varied , etc . the order or sequence of any process or method steps may be varied or re - sequenced according to alternative embodiments . in the claims , any means - plus - function clause is intended to cover the structures described herein as performing the recited function and not only structural equivalents but also equivalent structures . other substitutions , modifications , changes and omissions may be made in the design , operating conditions and arrangement of the preferred and other exemplary embodiments without departing from the spirit of the present inventions .

a tissue characterization apparatus and method may comprise a light source configured to direct light towards the tissue , a light receptor configured to receive light reflected from the tissue and configured to generate an electrical signal , and a spectrogram configured to analyze the signal generated by the light receptor .

the inventive sensor provides methods , apparatus and systems for optimizing the progression of a cardiac catheter into a blood flow extruding cardiac feature , such as through the coronary sinus . the difficulty with finding the coronary sinus lies in the fact that every heart is different in the spacing of anatomical features and in size and position of those features . therefore , the position of the coronary sinus typically varies from individual to individual . the bigger challenge in cardiac feature location within a clinical setting is that people with advanced heart failure typically have a much larger variation in their heart shape than that of the general population . in advanced heart failure , the heart enlarges , the coronary sinus tends to move down and toward the rv resulting in the expansion of the entire structure . the entire heart is enlarged and subsequently the location of the coronary sinus can be difficult to ascertain . to date most of the work that has been done with guiding catheters for coronary sinus access has been focused on the creation of custom shaped devices that will most likely locate the coronary sinus . these custom shapes are based on the average anatomy of the heart and therefore work for many instances where the anatomy is normal . this technique also works well in instances where coronary arteries are being located , for instance when placing stents and doing angioplasty . arteries are not blocked by valves at their openings and contrast injections can be used to find the entrance point for the catheter . the custom shapes in this environment serve two purposes : to find the artery and to keep the catheter engaged during the procedure . in the case of the coronary sinus the blood flow is coming toward the catheter and therefore the use of contrast dye is difficult and of little use and would require the injection of a large amount of dye . fig1 provides a depiction of the heart showing the coronary sinus 103 with blood coming out from it , as well as a catheter 100 with the inventive coronary sinus sensor . this coronary sinus sensor device is nicknamed the coronary sinus sniffing device or the coronary sinus location device . the coronary sinus sensor device is intended to be used in procedures where physicians are attempting to access the coronary sinus from the right atrium using a guiding catheter . this type of procedure is conducted when physicians are placing coronary sinus or venous leads for cardiac resynchronization therapy procedures . in these procedures , a third pacing lead is placed through the coronary sinus into the cardiac vein to allow pacing of the left ventricle . the invention in this disclosure is a coronary sinus guide that uses a different approach to the anatomy . rather than relying solely upon a special shape or combination of special shaped catheters to locate the coronary sinus , this invention utilizes the fact that blood is flowing out of the coronary sinus towards the catheter . by sensing this flow with some directionality , a catheter can be positioned in front of the coronary sinus to allow a guide or guide wire to be placed into the coronary sinus . the invention is the use of a flow sensor , such as a sensing wing , that allows the catheter to be positioned where the flow sensing wing senses the greatest amount of flow coming from the coronary sinus . fig2 a shows a guiding catheter 100 with a guide wire 101 ( as shown in fig3 b ) inserted and a flow sensing wing 102 at the tip of the device . arrows coming out of the coronary sinus 103 show the flow coming from the coronary sinus and deflecting the flow sensing wing . the flow sensing wing includes a strain sensing element as part of its design . therefore , when the front of the catheter is aligned with the coronary sinus , the wing is deflected by the blood flow , indicating that the catheter is aligned with the coronary sinus . fig2 b provides a view of a device similar to the embodiment shown in fig2 a with the exception that there is an opening or a side port 104 . side port 104 allows the blood flow from the coronary sinus to enter the catheter tip and then exit behind the wing . this feature may increase the effectiveness of the wing at sensing blood flow . fig2 c shows an embodiment similar to those in 2 a and in 2 b . in this configuration , catheter 100 and guide wire 101 are again provided . this view further provides flow sensing wing 102 in a different orientation . in this case , the blood flow comes into the guiding catheter 100 and flows out of the guiding catheter 100 through the side port 105 and as it flows out of the side port it deflects the wing 102 . fig2 d shows another embodiment of the design which includes an outer guide catheter 123 and inner guide catheter 106 with a guide wire 101 . the inner guide catheter 106 is deployed in front of the outer guide catheter 123 to protect the strain sensing wing 102 during insertion into the atrium . strain sensing wing 102 is shown hanging off the tip of the outer guide catheter 100 , as deployed for flow sensing . in the same way , this device allows sensing of the flow from the coronary sinus . when the catheter is aligned with the coronary sinus the flow sensing wing is deflected by the flow of blood . fig2 e shows the use a guiding catheter 100 with a guide wire 101 . this particular device has two flow sensing wings deployed within it . the first one , the distal flow sensing wing 102 is closest to the coronary sinus . distal flow sensing wing 102 is used to sense the coronary sinus flow . proximal flow sensing wing 107 is farthest away from the coronary sinus . proximal flow sensing wing 107 is used to sense other flow within the chamber , for instance flow from the inferior and superior vena cava . the blood flow comes into catheter 100 at the proximal wing 107 and exits the proximal side port 108 . in this way any flow from the inferior and superior vena cava that is sensed by the distal flow sensor wing 102 can be subtracted out so that only the flow recorded from the coronary sinus would be sensed . another use of this strain or flow sensing catheter is to allow the physician to slide the guide wire forward into the coronary sinus at the appropriate time . referring again to fig2 a , as the physician approaches the coronary sinus 103 the flow sensing wing will sense flow coming out of the coronary sinus 103 . the flow volume amplitude will typically have a signal that is periodic in nature somewhat like a sine wave . the blood flow data will indicate when the blood flow from the coronary sinus is at a maximum and a minimum . in many patients , there is a valve that covers the opening of the coronary sinus or a portion of the coronary sinus . this valve is termed the thebesian valve . the thebesian valve is like a skin flap which covers the opening to the coronary sinus . the thebesian valve can make entering the coronary sinus very difficult when a physician is using a normal guiding catheter . he or she has no way of knowing when this valve is open or closed . currently , clinicians rely on the catheter shape alone to get them in an area that is close to the valve . however , the clinician then has to relying on luck to slide the wire or the catheter into the coronary sinus at the appropriate time . the present inventive system allows , for the first time , the physician to know when the blood flow is at its maximum and therefore at which point the thebesian valve is open . if timed properly , the physician is now able to time the forward motion of the guide wire so that it enters the coronary sinus at just the time point that the flow is at the maximum , corresponding to the time point when the valve is open . this feature of the present invention will further enable physicians to access the coronary sinus more easily and more quickly in almost any clinical application regardless of the patient anatomy . fig3 a shows the use of a normally shaped guiding catheter 100 with a special guide wire or a flow sensing guide wire 109 that has a retractable thin membrane wing 111 . the concept embodied in this device is that the clinician slides a guide wire 109 down the guiding catheter 100 . when the guide wire 109 comes out of the tip of the device , the guide wire 109 has a tip 110 ( as shown in fig3 b ). when pulled backward in the guide wire 109 , tip 110 ( as shown in fig3 b ) deploys retractable wing 111 . fig3 b shows an enlarged view of guide wire 109 from fig3 a in its initial position for feeding through the catheter . in this view , the structure is shown tip 110 . strain sensing member 112 is also shown . fig3 c shows the guide wire 109 in the position with the wing 111 deployed . in this view , the guide wire tip 110 is pulled backwards towards the rest of the guide wire 109 in the direction of the arrow . in turn , guide wire 109 deploys thin membrane wing 111 . thin membrane wing 111 is surrounded by , for instance , two pieces of wire that form the desired shape . fig3 c further shows the guide wire wing 111 and its shape . the guide wire wing 111 also has a strain sensing member ( strain gauge ) 112 attached . in this manner , when the guide wire 109 is deflected by flow , because of the wing 111 , the strain sensing member 112 senses strain . this strain data is transmitted back as an indication that the catheter 100 is aligned with the coronary sinus , as shown in fig3 a . fig4 a and 4b show the catheter 100 approaching the coronary sinus 103 . the inventive method embodiment is conveyed in these figures . fig4 a shows that as the catheter 100 approaches the coronary sinus 103 , the flow wing 102 senses a little bit of the flow and therefore the deflection will be minimal . in fig4 b , as the catheter 100 becomes more aligned with the coronary sinus 103 , the flow sensing wing 102 will deflect more . this greater deflection as conveyed to the clinician from data received from strain gauges 112 is used as an indication that the catheter is well aligned with the coronary sinus 103 . fig4 c shows one embodiment of a wing design with a strain gauge 112 attached to the wing 102 design . in this embodiment , a large surface area is created by providing a paddle - like wing design . the blood flow from the coronary sinus deflects the strain sensor as a result of the wing design . the thinner area of the paddle on which strain gauge 112 is positioned is most likely to bend from that coronary sinus blood flow . referring to fig5 a , 5 b , and 5 c , these figures illustrate an additional design feature of the present invention for directing blood flow to optimize blood flow sensing . the feature is a catheter that is shaped in a manner that assists in the direction of blood flow . fig5 a shows an embodiment of the blood flow directing feature of the invention where the bottom lip 113 of the outer guide catheter 100 is extended so that the catheter 100 will catch and direct more of the blood flow towards the sensing wing 112 . this blood directing feature can be fabricated by providing an angled cut on the tubing . alternately , a tongue may be placed in the tip of the catheter which would scoop or direct blood into the guide catheter and direct it towards the flow sensing wing 112 . other device features to accomplish blood flow direction will be apparent to the skilled artisan . fig5 b shows an embodiment similar to that shown in fig5 a it with the side port 105 to allow blood to come in and go out of the catheter once it passes the flow sensing wing . fig5 c shows again the flow being directed into the catheter 100 and towards the flow sensing wing 112 . the blood flow will then come out of the catheter 100 just beyond the wing 112 arrows showing blood flow have been added for clarity . the above described embodiments of the present invention , as well as those apparent to the skilled artisan , can be employed with a number of different types of strain sensing devices . with the present invention , how the strain is measured , or how the flow is measured is not critical . rather , it is enough that measured flow is used to detect and locate the coronary sinus . typical strain gauges could be used , such as a piezoelectric device , fiber optics , semi conductor strain gauges , and the like . an example providing such strain gauges is found in the provisional patent applications “ cardiac motion characterization by strain measurement ” filed dec . 17 , 2004 by some of the present inventors , 60 / 638 , 247 , hereby incorporated by reference in its entirety . it is also important to note that with this particular device it may be valuable to include a pressure sensor at the tip of the device as well as the strain sensing device . the two devices in combination can be used to help begin the of the coronary sinus location process by measuring the local pressure within the right atrium . this pressure signal could also be used as a timing mechanism to time the introduction of a wire into the coronary sinus once it has been located by the coronary sinus location device of this invention . a prototype device similar to those show in fig2 a was constructed . in a water bath , a laminar flow of water was introduced from a tube to simulate flow from the coronary sinus . a periodic , moderate pulse in that liquid flow was provided . the prototype device was that directed in varying degrees towards this flow , simulating a catheter approaching the coronary sinus . when the sensor was directed towards , but somewhat obliquely towards the coronary sinus flow , the pressure sensor provide a moderate pressure sensor reaction , such as shown in fig4 a . as the simulated catheter more closely approached the fluid flow , the pressure sensor reaction was considerably increased , such as seen in fig4 b . the data obtained during this prototype experiment is provided in fig6 a & amp ; b .

the present invention provides a new mechanical sensor approach to maneuvering catheters and other cardiac devices into blood outlets , with particular application to maneuvering cardiac devices into the coronary sinus and beyond . additionally , the inventive sensing device provides assessment of the viability of branching veins and other potential device sites , such as within the coronary venous system .

referring to fig1 which is a side view of a liquid soap faucet dispenser ( 10 ) with a soap dispenser ( 14 ) in a horizontal position . the liquid soap faucet dispenser ( 10 ) comprises a faucet body ( 12 ). the faucet body ( 12 ) comprises a faucet adapter ( 12a ) positioned at a top distal end of the faucet body ( 12 ) and is swively connected , at one distal end , thereto . the opposite distal end of the faucet adapter ( 12a ) is adapted to connect the liquid soap faucet dispenser ( 10 ) to a standard sink faucet , which is a water source . the faucet body ( 12 ) further comprises a faucet water pressure reducer ( 12b ) sealably connected to the faucet adapter ( 12a ) extending downwardly therefrom within the faucet body ( 12 ). the faucet water pressure reducer ( 12b ) further comprises at least one faucet water pressure reducer side opening ( 12ba ) therein . the faucet water pressure reducer ( 12b ) further comprises at least one faucet water pressure reducer bottom opening ( 12bb ) therein . the faucet water pressure reducer ( 12b ) functions to reduce the water pressure flowing from the water source into the faucet body ( 12 ). the faucet body ( 12 ) further comprises a faucet connector ( 12c ) extending at a perpendicular angle outwardly therefrom . the faucet body ( 12 ) further comprises a faucet valve ( 12d ) positioned within a faucet valve sleeve ( 12da ) within the faucet connector ( 12c ). the faucet valve ( 12d ) comprises a faucet valve rod ( 12de ) longitudinally disposed in a central position within the faucet valve sleeve ( 12da ). the faucet valve ( 12d ) further comprises a faucet valve o - ring ( 12dj ) circumferentially positioned around the faucet valve sleeve ( 12da ) between the faucet connector ( 12c ) and the soap dispenser connector ( 14c ). the faucet valve o - ring ( 12dj ) functions to provide a seal between the faucet connector ( 12c ) and a soap dispenser connector ( 14c ). a faucet valve cap ( 12db ) is securely affixed to a first distal end of the faucet valve rod ( 12de ). a faucet valve seat ( 12dc ) and a faucet valve retainer ( 12dd ) are securely circumferentially positioned within the faucet valve sleeve ( 12da ). the faucet valve seat ( 12dc ) and the faucet valve retainer ( 12dd ) sealably engages the faucet valve cap ( 12db ) when in a closed position . a faucet valve first washer ( 12dga ) is positioned around the faucet valve rod ( 12de ) adjacent to the faucet valve seat ( 12dc ) and the faucet valve retainer ( 12dd ). a faucet valve spring ( 12df ) is positioned around the faucet valve rod ( 12de ) adjacent to the faucet valve first washer ( 12dga ). a faucet valve second washer ( 12dgb ) is positioned around the faucet valve rod ( 12de ) adjacent to the faucet valve spring ( 12df ). a faucet valve adjustment nut ( 12dh ) is treadably engaged on the faucet valve rod ( 12de ) adjacent to the faucet valve second washer ( 12dgb ). the faucet valve spring ( 12df ) functions to force the faucet valve seat ( 12dc ) against the faucet valve cap ( 12db ). the faucet valve adjustment nut ( 12dh ) functions to provide for adjustment of the force the faucet valve seat ( 12dc ) exerts against the faucet valve cap ( 12db ). the faucet valve ( 12d ) further comprises a faucet valve lock nut ( 12di ) which is threadably engaged on the faucet valve rod ( 12de ) positioned adjacent to the faucet valve adjustment nut ( 12dh ). the faucet valve lock nut ( 12di ), when tightened against the faucet valve adjustment nut ( 12dh ), functions to prevent the faucet valve adjustment nut ( 12dh ) from rotating away from a preselected position . the liquid soap faucet dispenser ( 10 ) further comprises a soap dispenser ( 14 ) which comprises a soap dispenser adapter ( 14a ) positioned at a top distal end . the soap dispenser adapter ( 14a ) comprises a soap dispenser adapter soap adjuster ( 14aa ) functioning to regulate an amount of liquid soap dispensed therefrom , soap dispenser soap tube ( 14b ), having a soap dispenser soap tube outlet ( 14ba ), is sealably connected to the soap dispenser adapter ( 14a ) extending downwardly therefrom through an opening in the soap dispenser ( 14 ), a bottom end of a soap dispenser reservoir ( 14j ) is sealably connected to the soap dispenser adapter ( 14a ). the soap dispenser reservoir ( 14j ) comprises a soap dispenser reservoir opening ( 14ja ) at a top end thereof , a soap dispenser reservoir opening cap ( 14jaa ) is removably sealably positioned on the soap dispenser reservoir opening ( 14ja ). the soap dispenser reservoir opening cap ( 14jaa ) comprises a soap dispenser air release valve ( 14jaaa ). the soap dispenser air release valve ( 14jaaa ) functions as a vacuum break . the soap dispenser ( 14 ) further comprises a soap dispenser connector ( 14c ) extending perpendicularly therefrom rotatably mounted on the faucet valve sleeve ( 12da ). the faucet valve rod ( 12de ) extends longitudinally in a middle of the soap dispenser connector ( 14c ) and terminates in a soap dispenser end sleeve ( 14h ) securely mounted to an inside wall of the soap dispenser ( 14 ). the soap dispenser connector ( 14c ) further comprises a slanted soap dispenser connector slot ( 14ca ) therein , a soap dispenser connector in ( 14d ) is securely attached to the faucet valve sleeve ( 12da ) within the soap dispenser connector slot ( 14ca ), a soap dispenser washer ( 14g ) is positioned around the faucet valve rod ( 12de ) adjacent to the soap dispenser end sleeve ( 14h ), a soap dispenser lock nut ( 14e ) is threadably mounted on the faucet valve rod ( 12de ) adjacent to the soap dispenser lock nut ( 14e ). a soap dispenser adjustment nut ( 14f ) is threadably positioned on the faucet valve rod ( 12de ) adjacent to the soap dispenser lock nut ( 14e ). the soap dispenser adjustment nut ( 14f ) functions when tightened against the soap dispenser lock nut ( 14e ) to prevent the soap dispenser lock nut ( 14e ) from rotating away from a preselected position . the soap dispenser ( 14 ) further comprises a soap dispenser outlet ( 14i ) positioned on a side thereof when a user positions the soap dispenser ( 14 ) in a horizontal position , the faucet valve spring ( 12df ) compresses which causes the faucet valve cap ( 12db ) to disengage from the faucet valve seat ( 12dc ) allowing water to flow from the faucet through the faucet body ( 12 ) and the soap dispenser ( 14 ) egressing through the soap dispenser outlet ( 14i ). when the user positions the soap dispenser ( 14 ) in a vertical position , the faucet valve spring ( 12df ) decompresses causing the faucet valve cap ( 12db ) move toward and engage the faucet valve seat ( 12dc ) which stops the flow of water and allows liquid soap to flow from the soap dispenser reservoir ( 14j ) through the soap dispenser soap tube ( 14b ) egressing from the soap dispenser soap tube outlet ( 14ba ). lastly , referring to fig2 which is a side view of the liquid soap faucet dispenser ( 10 ) with the soap dispenser ( 14 ) in a vertical position . the liquid soap faucet dispenser ( 10 ) comprises a faucet body ( 12 ). the faucet body ( 12 ) comprises a faucet adapter ( 12a ) positioned at a top distal end of the faucet body ( 12 ) and is swively connected , at one distal end , thereto . the opposite distal end of the faucet adapter ( 12a ) is adapted to connect the liquid soap faucet dispenser ( 10 ) to a standard sink faucet , which is a water source . the faucet body ( 12 ) further comprises a faucet water pressure reducer ( 12b ) sealably connected to the faucet adapter ( 12a ) extending downwardly therefrom within the faucet body ( 12 ). the faucet water pressure reducer ( 12b ) further comprises at least one faucet water pressure reducer side opening ( 12ba ) therein . the faucet water pressure reducer ( 12b ) further comprises at least one faucet water pressure reducer bottom opening ( 12bb ) therein . the faucet water pressure reducer ( 12b ) functions to reduce the water pressure flowing from the water source into the faucet body ( 12 ). the faucet body ( 12 ) further comprises a faucet connector ( 12c ) extending at a perpendicular angle outwardly therefrom . the faucet body ( 12 ) further comprises a faucet valve ( 12d ) positioned within a faucet valve sleeve ( 12da ) within the faucet connector ( 12c ). the faucet valve ( 12d ) comprises a faucet valve rod ( 12de ) longitudinally disposed in a central position within the faucet valve sleeve ( 12da ). the faucet valve ( 12d ) further comprises a faucet valve o - ring ( 12dj ) circumferentially positioned around the faucet valve sleeve ( 12da ) between the faucet connector ( 12c ) and the soap dispenser connector ( 14c ). the faucet valve o - ring ( 12dj ) functions to provide a seal between the faucet connector ( 12c ) and a soap dispenser connector ( 14c ). a faucet valve cap ( 12db ) is securely affixed to a first distal end of the faucet valve rod ( 12de ). a faucet valve seat ( 12dc ) and a faucet valve retainer ( 12dd ) are securely circumferentially positioned within the faucet valve sleeve ( 12da ). the faucet valve seat ( 12dc ) and the faucet valve retainer ( 12dd ) sealably engages the faucet valve cap ( 12db ) when in a closed position . a faucet valve first washer ( 12dga ) is positioned around the faucet valve rod ( 12de ) adjacent to the faucet valve seat ( 12dc ) and the faucet valve retainer ( 12dd ). a faucet valve spring ( 12df ) is positioned around the faucet valve rod ( 12de ) adjacent to the faucet valve first washer ( 12dga ). a faucet valve second washer ( 12dgb ) is positioned around the faucet valve rod ( 12de ) adjacent to the faucet valve spring ( 12df ). a faucet valve adjustment nut ( 12dh ) is treadably engaged on the faucet valve rod ( 12de ) adjacent to the faucet valve second washer ( 12dgb ). the faucet valve spring ( 12df ) functions to force the faucet valve seat ( 12dc ) against the faucet valve cap ( 12db ). the faucet valve adjustment nut ( 12dh ) functions to provide for adjustment of the force the faucet valve seat ( 12dc ) exerts against the faucet valve cap ( 12db ). the faucet valve ( 12d ) further comprises a faucet valve lock nut ( 12di ) which is threadably engaged on the faucet valve rod ( 12de ) positioned adjacent to the faucet valve adjustment nut ( 12dh ). the faucet valve lock nut ( 12di ), when tightened against the faucet valve adjustment nut ( 12dh ), functions to prevent the faucet valve adjustment nut ( 12dh ) from rotating away from a preselected position . the liquid soap faucet dispenser ( 10 ) further comprises a soap dispenser ( 14 ) which comprises a soap dispenser adapter ( 14a ) positioned at a top distal end . the soap dispenser adapter ( 14a ) comprises a soap dispenser adapter soap adjuster ( 14aa ) functioning to regulate an amount of liquid soap dispensed therefrom , soap dispenser soap tube ( 14b ), having a soap dispenser soap tube outlet ( 14ba ), is sealably connected to the soap dispenser adapter ( 14a ) extending downwardly therefrom through an opening in the soap dispenser ( 14 ), a bottom end of a soap dispenser reservoir ( 14j ) is sealably connected to the soap dispenser adapter ( 14a ). the soap dispenser reservoir ( 14j ) comprises a soap dispenser reservoir opening ( 14ja ) at a top end thereof , a soap dispenser reservoir opening cap ( 14jaa ) is removably sealably positioned on the soap dispenser reservoir opening ( 14ja ). the soap dispenser reservoir opening cap ( 14jaa ) comprises a soap dispenser air release valve ( 14jaaa ). the soap dispenser air release valve ( 14jaaa ) functions as a vacuum break . the soap dispenser ( 14 ) further comprises a soap dispenser connector ( 14c ) extending perpendicularly therefrom rotatably mounted on the faucet valve sleeve ( 12da ). the faucet valve rod ( 12de ) extends longitudinally in a middle of the soap dispenser connector ( 14c ) and terminates in a soap dispenser end sleeve ( 14h ) securely mounted to an inside wall of the soap dispenser ( 14 ). the soap dispenser connector ( 14c ) further comprises a slanted soap dispenser connector slot ( 14ca ) therein , a soap dispenser connector pin ( 14d ) is securely attached to the faucet valve sleeve ( 12da ) within the soap dispenser connector slot ( 14ca ), a soap dispenser washer ( 14g ) is positioned around the faucet valve rod ( 12de ) adjacent to the soap dispenser end sleeve ( 14h ), a soap dispenser lock nut ( 14e ) is threadbly mounted on the faucet valve rod ( 12de ) adjacent to the soap dispenser lock nut ( 14e ). a soap dispenser adjustment nut ( 14f ) is threadbly positioned on the faucet valve rod ( 12de ) adjacent to the soap dispenser lock nut ( 14e ). the soap dispenser adjustment nut ( 14f ) functions when tightened against the soap dispenser lock nut ( 14e ) to prevent the soap dispenser lock nut ( 14e ) from rotating away from a preselected position . the soap dispenser ( 14 ) further comprises a soap dispenser outlet ( 14i ) positioned on a side thereof when a user positions the soap dispenser ( 14 ) in a horizontal position , the faucet valve spring ( 12df ) compresses which causes the faucet valve cap ( 12db ) to disengage from the faucet valve seat ( 12dc ) allowing water to flow from the faucet through the faucet body ( 12 ) and the soap dispenser ( 14 ) exiting through the soap dispenser outlet ( 14i ). when the user positions the soap dispenser ( 14 ) in a vertical position , the faucet valve spring ( 12df ) decompresses causing the faucet valve cap ( 12db ) move toward and engage the faucet valve seat ( 12dc ) which stops the flow of water and allows liquid soap to flow from the soap dispenser reservoir ( 14j ) through the soap dispenser soap tube ( 14b ) exiting from the soap dispenser soap tube outlet ( 14ba ). it will be understood that each of the elements described above , or two or more together , may also find an useful application in other types of constructions differing from the type described above . while the invention has been illustrated and described as embodied in a liquid soap faucet dispenser , it is not intended to be limited to the details shown , since it will be understood that various omissions , modifications , substitutions and changes in the forms and details of the device illustrated and in its operation can be made by those skilled in the art without departing in any way from the spirit of the present invention . without further analysis , the foregoing will so fully reveal the gist of the present invention that others can , by applying current knowledge , readily adapt it for various applications without omitting features that , from the standpoint of prior art , fairly constitute essential characteristics of the generic or specific aspects of this invention . what is claimed as new and desired to be protected by letters patent is set forth in the appended claims .

a device which is attached to the end of a standard kitchen faucet and dispenses liquid soap on to an object to be washed . when dispensing soap the device cuts off the flow of water to permit the user and opportunity to wash the object and conserve water . the device is activated by rotating the device in a first direction which cuts off the flow of water and dispensed liquid soap . when the device is returned to the original position the water flow is restored and the dispensing of liquid soap is stopped .

with reference now to the drawings , and in particular to fig1 and 4 thereof , the preferred embodiment of the new and improved device for an eversion assist of inverted nipples embodying the principles and concepts of the present invention and generally designated by the reference numeral 10 will be described . the present invention , the device for an eversion assist of inverted nipples 10 , is comprised of a plurality of components . such components in their broadest context include upper and lower surfaces , a central aperture , and an exterior periphery . such components are individually configured and correlated with respect to each other so as to attain the desired objective . more specifically , the present invention is a uniquely “ user friendly ” apparatus of ultralight materials comprising a multi - layered single - unit construction with adhesive back . the device of the present invention includes a substantially annular raised composite with aperture which fits over the areola and relies upon the interplay between the device &# 39 ; s flat base , the adhesive and the areola tissue to manipulate the nipple orientation and maintain a protracted state . unlike products comprising the prior art , many of which utilize suction , vacuum and are often cumbersome methods / devices , the present “ user friendly ” invention effectively treats inverted nipples by means of a continuous gentle , yet focused nudge effect achieved when the adhesive - back flat base of the device meets with the areola tissue . further benefits of this new and improved solution for treating inverted nipples relate to the device &# 39 ; s ultralight construction and adhering properties , which make this device easy to apply , comfortable to wear for extended periods , capable of remaining securely in place while active or asleep , as well as affording improved concealability over prior art , which can be particularly advantageous when only one nipple is inverted . adding to its “ user - friendly ” attributes , this self - adhering ultralight device can be configured to be efficiently manufactured for disposable use , including peel - and - stick properties , thus eliminating the need for maintenance . the present invention comprises a 3 - dimensional , substantially annular , multi - tiered composite areolar device measuring from 0 . 25 ″- 1 ″, ( 6 . 35 mm - 25 . 4 mm ) thickness . the topcoat is comprised of a thin membrane of latex , felt , moleskin , silicone or other lightweight polymer . the topcoat is bonded to an inner layer of ultralight material to provide firmness and cushioning . this ultralight material substance is firm enough to allow the device to be firmly pressed against the surface of the areola . the flat bottom layer is a self - sticking medical grade adhesive that is bonded to the bottom external layer of the device . this self - sticking adhesive is characterized as having : a tackiness of 10 - 450 grams as determined by a cheminstruments polyken probe tack tester and a tensile strength of 0 . 14 - 5 . 52 mega pascals ( 20 - 800 pounds / square inch ), a minimum elongation of 250 - 1100 percent and a tear strength of 0 . 88 - 35 . 2 kn / m , 5 - 200 pounds / square inch . the overall diameter of the device measures between 1 ″- 4 ″, ( 25 . 4 mm - 101 . 1 mm ) diameter with a center aperture between 0 . 25 ″- 1 ″, ( 6 . 35 mm - 25 . 4 mm ) that are manufactured into rings to conform to various areola and nipple sizes . specifically , the device is designed to address the condition presented by grade 1 and grade 2 inverted nipples , i . e . those protractible with light inward pressure to the areola . this “ user - friendly ” single - unit device can be applied with little time or effort involved . first , by using a method that is most comfortable to the user , the nipple is protracted . if the nipple cannot be everted easily , a doctor should be consulted . second , remove the protective backing to expose the adhesive and apply the device so the nipple protrudes through the center opening , then gently press onto the areola . the affected result is that the device applies a slight pressure inward and upward to the areola tissue around the nipple base creating a continuous gentle , yet focused nudge - effect , while also keeping the areola tissue from spreading and thereby allowing for possible tightening around the nipple base which may prove beneficial for lasting results . once applied , the device may be worn comfortably and securely , with or without clothing , for extended periods of time . with continued use , results can vary , to include long lasting protraction in some users . because of the small size and ultralight design of this device , one can expect the least amount of discomfort or intrusion upon the natural freedom of the breast and nipple . several attributes include its easy of application and use . the device may be worn comfortable for extended periods of time . the device is light weight and nearly invisible under clothing . the device need not rely on suction , creams , multiple parts , or piercing , but instead may rely on an ultra - light adhesive backing . the device does not require obtrusive hardware . nor does the device require on - going cleaning or sterilization . the present invention is a device 10 for an eversion assist of inverted nipples . the device is fabricated of ultralight material and has a generally three - dimensional annular configuration with an upper layer or upper planar surface 14 and a parallel lower planar surface 16 . a cylindrical aperture 28 extends centrally through the device . the central aperture has an interior periphery or interior surface of the aperature 30 in a cylindrical configuration extending between the lower planar surface and the upper planar surface . the cylindrical aperture has a second central axis co - extensive with the first central axis . the cylindrical aperture generally has a diameter of 25 percent ( 25 %) of the lower diameter . the lower planar surface 16 has a radially interior edge or aperture opening 18 and a radially exterior edge 20 . the upper planar surface 14 has a radially interior edge 22 and a radially exterior edge 24 . the device has a first central axis . the lower peripheral surface has a lower diameter of from 1 to 4 inches . the upper planar surface has a diameter of from 70 to 100 percent ( 70 %- 100 %) of the lower diameter . the device generally has a height of from 12 . 5 to 25 percent ( 12 . 5 %- 25 %) of the lower diameter . the device has a lower exterior periphery 36 in a cylindrical configuration extending upwardly from the radial exterior edge of the lower planar surface and terminating at an intermediate circle 44 . the lower exterior periphery has a peripheral height of from 65 to 100 percent ( 65 %- 100 %) of the height of the device . the device may have an upper exterior periphery 40 in a frusto - conical configuration extending upwardly from the intermediate circle and terminating at the radially exterior edge of the upper planar surface . an upper layer 14 of a cover material is adhered to the upper surface and the upper exterior periphery . the upper layer is comprised of a thin membrane chosen from the class comprised of latex , elastomer , felt , moleskin , silicone , or other lightweight polymer . a lower layer 48 is a self - sticking tacky adhesive or adhesive layer adhered to the lower surface . the lower layer is comprised of a medical grade adhesive . as to the manner of usage and operation of the present invention , the same should be apparent from the above description . accordingly , no further discussion relating to the manner of usage and operation will be provided . with respect to the above description then , it is to be realized that the optimum dimensional relationships for the parts of the invention , to include variations in size , materials , shape , form , function and manner of operation , assembly and use , are deemed readily apparent and obvious to one skilled in the art , and all equivalent relationships to those illustrated in the drawings and described in the specification are intended to be encompassed by the present invention . therefore , the foregoing is considered as illustrative only of the principles of the invention . further , since numerous modifications and changes will readily occur to those skilled in the art , it is not desired to limit the invention to the exact construction and operation shown and described , and accordingly , all suitable modifications and equivalents may be resorted to , falling within the scope of the invention . the device can be described as a combination of other features as well . the device 10 may be a single - unit mold or a composite of multiple plies adhered together . the device may be made of a latex layer or be entirely latex - free . the device may be made of a synthetic or natural polymer . the device may be made of poron . the diameter of the device may be from 1 . 5 to 4 inches . the thickness of the device may be from 0 . 125 - 1 inches . the device may comprise an upper layer 14 , an adhesive layer 48 , and an aperture 28 , as shown in fig3 and 4 . the device may also feature a substantially flat portion which may be conterminous with the lower planar surface 16 . the exterior , which may be conterminous with lower exerior periphery 36 , the upper exerior periphery 40 , and the upper layer 14 , may have a straight , tapered , spherical , conical , frustoconical , angular edge , polygonal , or ovoid shape . the upper layer may feature a substantially flat upper surface and a flat lower surface , which may be the same as the lower planar surface 16 , and may be made of felt , moleskin , foam , latex , elastomer , silicone , another flexible polymer , or a combination of these materials . the upper layer may serve as an exterior of the device . the lower surface of the upper layer may be disposed flush against the adhesive layer . the adhesive layer may be configured to adhere to an areola tissue to manipulate nipple orientation . specifically , it may be configured to adhere at least a portion of the areola tissue into a position parallel to the substantially flat portion . it may be formed from woven fabric , and / or feature a plastic portion or latex portion . the adhesive layer may be protected by a protective back material ( not shown ). this protective back material may be removably attached to the adhesive layer . the protective back material may feature a tag enabling easy removal . the aperture may be disposed centrally in the device . it may be contoured to receive a nipple when at least a portion of the areola tissue is oriented in a position parallel to the substantially flat portion . the configuration of the aperture may be such that the nipple may enter though a gap in the adhesive layer . the aperture may be cylindrical , conical , frustoconical , or spherical in shape . the inner surface of the aperture 30 may feature a continuation of the adhesive layer or the adhesive layer may terminate at or near the opening of the aperture 18 . the diameter of the aperture may range from 0 . 25 to 1 inch .

the invention is a single - unit areolar device of ultralight construction with an adhesive back that relies on no additional parts or components to effectively manipulate nipple orientation for eversion assist . the device builds upon the interplay between the adhesive - back flat base with the areola tissue to create a gentle , focused nudge to evert assist and maintain protraction of inverted nipples . the ultralight single - unit device is comfortable to wear for extended periods of time , easily concealed beneath clothing , and remains securely in place while active or asleep . the disposable attributes of the cost effective materials utilized lend further advantage to the device .

as shown in fig1 - 3 , our invention is installed in a rotary blade lawnmower 10 including a housing 11 in which a blade 12 is rotated about a vertical axis . as usual , housing 11 is mounted on wheels 13 and is provided with a handle 14 including a throttle control 15 for the engine 16 , shown schematically , which is mounted on the housing as by fasteners 17 . fig4 and 5 show that engine 16 has a vertical shaft 20 extending downwardly . housing 11 has two further portions 21 and 22 to contain mechanism about to be described : these portions are held together by fasteners 23 , and the assembly is mounted to the main housing by fasteners 17 . main housing 11 and portion 21 are centrally apertured to pass shaft 20 , and portion 22 is also centrally apertured . portions 21 and 22 cooperate to define a clutch chamber 24 containing a driving member 25 secured to shaft 20 by means of a woodruff key 26 and passing through an oil seal 27 in portion 21 . a driven member 30 is also mounted in chamber 24 , on a bearing 31 , and passes through a second oil seal 32 . a limited amount of axial movement of member 30 in bearing 31 is possible . a driver 33 for blade 12 is secured to member 30 by means including a plate 34 and a bolt 35 . driven member 30 is bored at 36 to act as a pilot bearing for shaft 20 , and has a central annular ridge 37 . members 25 and 30 have flat inward surfaces 40 and 41 respectively , and a disc 42 of friction material is located therebetween , being centered by ridge 37 . a thrust bearing 43 is located between the outward upper surface 44 of member 25 and the adjacent wall of housing portion 21 , and a disc spring 45 is located between the outward lower surface 46 of driven member 30 and bearing 31 . the arrangement is such that spring 45 continuously urges member 30 upwardly to cause engagement of surfaces 40 and 41 with disc 42 , biasing the structure into a configuration which causes power transmission from shaft 20 to blade driver 33 . driven member 30 is of larger diameter than driving member 25 , and a further friction ring 50 rests on surface 41 and surrounds disc 42 . ring 50 comprises part of a control means 51 which also includes a flat metal ring 52 engaging the upper surface of friction ring 50 . the upper surface of ring 52 is provided with a plurality of grooves 53 of limited arcuate extent disposed circularly about the axis of shaft 20 and of uniformly varying depths . a like plurality of steel balls 54 ride in grooves 53 . a second set of similar grooves 55 is formed in a second flat ring 56 fixed to housing portion 21 in apposition to ring 52 , or these may indeed be formed directly in the housing portion . ring 56 and housing portion 21 are formed with aligned arcuate slots 57 , 60 and a pin 61 projects from ring 52 through the arcuate slots . a lever 62 pivoted at 63 has a slot 64 at its inner end to engage pin 61 , and is restrained under a cover plate 65 . at the outer end of lever 62 a tab 66 projects upwardly through a slot 67 in housing 11 , and carries a connector 70 around which is passed one looped end 71 of a coil spring 72 having its other end 73 secured to housing 11 as at 74 . also secured to connector 70 by a set screw 75 is the end of a bowden wire 76 , whose outer spiral 77 is clamped to housing 11 at 80 . we have found that tempered masonite is an excellent material for use as disc 42 and ring 50 , particularly if the chamber 20 is filled with oil . disc 42 has a plurality of apertures 81 , and driving member 25 has a plurality of apertures 82 , to allow the egress of oil trapped between the disc and the flat surfaces when power transmission is being initiated . grooves 53 and 55 have their varying depths increasing in opposite directions , grooves 53 being deeper in the direction of rotation of shaft 20 , and grooves 55 being shallower in that direction . thus , when ring 52 is pivoted clockwise as seen in fig5 with respect to ring 56 , the rings are displaced axially by twice the depth difference of a groove . this is accompanied by downward movement of ring 52 , friction ring 50 , and driven member 30 , which is permitted this slight axial movement in bearing 31 . lever motion is caused by the action of coil spring 72 which overrides disc spring 45 : power transmission to drive member 30 is interrupted at disc 42 , and the member is braked at ring 50 . this is the normal or inoperative state of the device . when it is desired to operate the mower , the user actuates a &# 34 ; deadman &# 34 ; type of control handle 83 , acting through bowden wire 76 to pivot lever 62 clockwise as seen in fig5 . this allows rings 52 and 56 to take the relative positions shown in fig6 where they are at their closest spacing . now no significant amount of force is applied through balls 54 , flat ring 52 and fixed ring 50 to driven member 30 : disc spring 45 urges driven member 30 toward member 25 to cause engagement between the members and disc 42 , any intervening oil being forced out either at the edge of the disc or through the apertures 81 , 82 , and power transmission is established . it is to be particularly noted that when handle 83 is released , spring 72 begins to act through lever 62 on ring 51 , and that the direction of rotation given to ring 52 by ring 50 and member 30 is such as to urge the ring rapidly in the braking direction . this accomplishes full braking even should spring 72 weaken to give less than rated force when fully contracted . fig8 shows a modification of the invention in which shaft 120 is connected to driving member 125 by an overriding clutch including balls 129 instead of by a key . in this arrangement , blade 12 may be manually rotated forward , without also driving shaft 20 and engine 16 , which is sometimes an advantage . a second modification of the invention is shown in fig9 where parts which are the same as in the earlier modification are given the same numbers in the 200 series . this embodiment differs from that of fig4 in the provision of an additional friction ring 249 secured in housing portion 222 below driven member 230 . in the driving condition of the apparatus , this new ring is not operative , but when ring 252 is rotated to force ring 250 and driven member 230 downward , the outward lower surface 246 of the latter moves into engagement with ring 249 , thus multiplying the friction area and hence the braking action . a further modification of the invention is shown in fig1 , wherein parts which are the same in earlier modifications are given the same numbers in the 300 series . in this embodiment the clutch - brake mechanism is not intended to operate in oil , and is not contained in housing elements such as 21 , 22 or fig1 . housing 311 has a much larger central opening 318 than before , and motor 316 has a ridge 319 projecting through opening 318 which takes the place of member 56 of fig4 and carries on its lower surface three curved grooves 355 of uniformly varying depth and limited angular extent to receive balls 354 . an annular member 351 carries similar grooves 353 to receive balls 354 in cooperation with grooves 355 , whereby to give the member limited rotation about the axis of shaft 320 , and may be actuated by extension 368 carrying an actuator connection 370 . motor shaft 320 is provided with shoulder 328 at a specific distance from the end of the shaft , and a driving member 344 is carried on the shaft and driven by a key 326 . a driven member 330 is carried in a bearing 331 on shaft 320 , and a pair of disc springs 345 act between bearing 331 and a lower surface 346 of member 330 . a single disc 348 of friction material is located between driven member 330 and driving member 344 , and performs the functions of members 42 and 50 of fig4 . the elements just recited are held in place by a washer 338 and bolt 335 . blade 312 is bored at 339 to pass the head of bolt 335 , and is mounted on member 330 by bolts 329 . as before , grooves 353 and 355 are of oppositely varying depths , so that member 351 moves downwardly , as seen in fig1 , with rotation of the member in the same direction as that of shaft 320 , to brake member 330 , through friction member 348 , and release the drive . the distance between shoulder 328 and the end of shaft 320 is such that when bolt 335 is tightened , springs 345 are put under considerable stress , and press driven member 330 and hence member 348 toward driving member 344 to make a driving connection from motor 316 to blade 312 . in the normal rotated position of the member 351 , however , it is forced by balls 354 against friction member 348 , compressing springs 345 to release the drive engagement with member 344 and to brake member 330 by friction through member 348 . operation of actuator 368 to rotate member 351 in the opposite direction allows springs 345 to push members 330 and 348 into driving engagement with member 344 and release the brake and allow slip between members 348 and 351 . if one of members 330 and 351 is bronze and the other is steel , the interposition of a separate friction disc 348 may not be necessary , the natural surface characteristics of these dissimilar metals being sufficient friction means to accomplish the desired clutching and braking satisfactorily . fig1 - 14 show how our inventive principle may be applied to the simple transmission of power from a drive shaft 401 to a driven shaft 402 , carried respectively in pillow blocks 403 and 404 . a driving member 405 is carried on shaft 401 and driven through a key 406 . a shoulder 407 is provided on shaft 401 against which is positioned a first bearing 410 carrying a first control member 411 from which a positioning pin 412 projects into an opening 413 in pillow block 403 , to prevent rotation of the member . a second , annular control member 414 is concentric with and spaced from member 411 . three balls 415 are carried between short circular grooves 416 in member 411 and similar grooves 417 in member 414 : the grooves are of opposite uniformly varying depth , and the arrangement is the same as that of elements 54 , 55 , and 53 of fig6 . separated from members 414 and 405 by a disc of friction material 420 carried by shaft 401 is a driven member 421 carried on shaft 401 in a bearing 422 received in a recess 423 in the member : a pair of disc springs 424 act between bearing 422 and the bottom surface 425 of recess 423 . the elements are held in assembled relation by washer 426 and bolt 427 . if the load on shaft 402 has considerable inertia , friction disc 420 may be made of material known as raybestos r451 , a product of raybestos - manhatten incorporated , trumbull , connecticut . memeber 411 has a peripheral notch 430 through which projects a connection 431 carried by member 414 to permit angular displacement of member 414 with respect to member 411 by suitable means such as a bowden wire 432 . as in fig1 , the distance between shoulder 407 and the end of shaft 401 is such that when bolt 427 is tightened a frictional drive occurs between members 405 and 421 through disc 420 , unless member 414 is rotated to press against disc 414 toward member 421 , compressing springs 424 , releasing the friction drive from member 405 , and braking member 421 . the left - hand end of member 421 is formed as one - half 433 of a dog clutch 434 , of which the other half 435 is secured to or integral with shaft 402 . this provides for the slight axial movement of member 421 which accompanies control operation of member 414 . from the foregoing it will be evident that we have invented a new and improved arrangement which combines clutching and braking operations as opposite states of a single simple mechanism , and one moreover in which the motion of the member being braked has itself the effect of maximizing the braking force . numerous characteristics and advantages of our invention have been set forth in the foregoing description , together with details of the structure and function of the invention , and the novel features thereof are pointed out in the appended claims . the disclosure , however , is illustrative only , and changes may be made in detail , especially in matters of shape , size , and arrangement of parts , within the principle of the invention , to the full extent indicated by the broad general meaning of the terms in which the appended claims are expressed .

a combined clutch and brake mechanism for use in such applications as power lawnmowers . during actuation of a manual control the brake is released and power is transmitted from the engine to the blade carrier . if the manual control is released , power transmission is disabled and braking is initiated , the braking being maximized by the very motion of the member being braked . the invention is also shown in a form adaptable for use in general power transmission applications .

the following detailed description is of the best currently contemplated modes of carrying out exemplary embodiments of the invention . the description is not to be taken in a limiting sense , but is made merely for the purpose of illustrating the general principles of the invention , since the scope of the invention is best defined by the appended claims . various inventive features are described below that can each be used independently of one another or in combination with other features . broadly , embodiments of the present invention generally provide a self - locking elastomeric loop that may be easily engaged and released from knobs or handles of cabinets and / or appliances by an adult , but which may be difficult to remove by a small child . referring now to fig1 and 2 it may be seen that an exemplary embodiment of a self - locking device 10 may include a closed loop 12 and one or more locking balls 14 formed outside of the loop 12 . the device 10 may be constructed from an elastomeric material such as silicone rubber . advantageously the device 10 may be constructed from material that may have regulatory approval for contact by small children . the loop 12 and locking balls 14 may constructed as a single integrally molded device . the loop 12 may be constructed with a cross - sectional thickness between about 3 mm to about 7 mm . the device 10 may be constructed in various sizes with lengths of the loop 12 ranging from about 60 mm to about 150 mm . advantageously , the loop 12 may have a teardrop shape . the balls 14 may be spherical and may have a diameter between about 10 mm to about 25 mm . the balls 14 may be positioned at ends of extensions 16 at a distance of about 10 mm to about 20 mm away from a joining point 18 of the loop 12 . in an exemplary configuration , a first one of the extensions 16 may project outwardly from the joining point 18 in alignment with a right hand portion 12 - 1 of the loop 12 and a second one of the extensions 16 may project outwardly from the joining point 18 in alignment with a left hand portion 12 - 2 of the loop 12 . referring now to fig3 and 4 , it may be seen that , in an exemplary application , that the device 10 may be placed around two handles 40 of a refrigerator 42 . the loop 12 may be stretched and wrapped around the handles so that a top end 20 overlies a bottom end 21 of the loop 12 . the top end 20 of the loop 12 may be passed over the locking balls 14 and then allowed to relax into engagement with the extensions 16 . in this configuration the device 10 may be considered to be in a locked mode . the locking balls 14 may prevent the device 10 from opening . thus the refrigerator doors 40 may be held closed by the device 10 in the event that a child attempts to open the refrigerator 42 . an adult may easily open the refrigerator 42 by pulling the top end 20 of the loop 12 over the locking balls 14 to release the device 10 . the adult may just as easily replace the device 10 on the handles 40 after closing the refrigerator 42 . referring now to fig5 , 6 and 7 , it may be seen the device 10 may be employed in various other exemplary applications . as seen in fig5 and 7 , a holding hook 30 may be adhesively secured to an interior surface of a cabinet 32 . the device 10 may be placed over the hook 30 and then stretched around a drawer face 34 to engage with a cabinet knob 26 . alternatively , a wrap - around hook 31 , as shown in fig9 , may be employed to secure the device 10 within the cabinet 32 . it may be seen that the devices 10 may be stretched from a first knob 26 - 1 to a second knob 26 - 1 while being passed behind a stile 38 of a cabinet . alternatively , one the devices 10 may be placed onto one of the hooks 30 and stretched around one of the knobs 26 of a cabinet door 36 . one or more of the devices 10 and one or more of the hooks 30 may be combined and sold as a kit 50 . one of the kits 50 may contain a plurality of the devices 10 which may have various lengths ranging from about 6 inches to about 12 inches . with such a kit , a parent of a young child may be able to secure cabinet doors and drawers of various sizes and configurations . it should be understood , of course , that the foregoing relates to exemplary embodiments of the invention and that modifications may be made without departing from the spirit and scope of the invention as set forth in the following claims .

a child - proof safety locking system may include an elastomeric loop . the loop may have at least one locking ball attached at a bottom end of the loop so that when the loop is wrapped around an object , a top end of the loop surrounds the locking ball and the top end of the loop is held in position overlying the bottom end of the loop .

a chair having adjustable arms according to the invention is generally shown in drawing fig1 . the chair has a base 100 , a seat 102 supported by the base 100 , a back 104 connected with the base 100 , typically two arms 106 connected with the base 100 , and an arm adjustment device 110 interposed between the arms 106 and the base 100 . the arm adjustment device 110 is generally shown throughout the drawing figures , and further has a drive rod 112 , a frame 114 adapted to hold the drive rod 112 in rotatable engagement , a torque member 116 operatively connected between the frame 114 and the drive rod 112 , a brake 118 operatively connected between the frame 114 and the drive rod 112 , and a brake actuator 120 operatively connected with the brake 118 . one having ordinary skill in the art will appreciate the fact that an arm adjustment device of the invention may be implemented in numerous embodiments , depending upon the preferences of the user . one , exemplary embodiment is generally shown in the drawing figures , generally identified by reference number 110 , and more specifically discussed below . in this embodiment , a support extends generally transversely across the chair back to support the arm adjustment device 110 and a pair of arms 106 on a front side of the support . the drive rod 112 is shown as a straight , one - piece member that interconnects the two arms 106 at rearward ends of the arms . in possible alternative embodiments , an arm adjustment device may be mounted on a back side of the support . also , the support might extend arcuately across the chair and the drive rod 112 may be a multi - segmented member with the segments being articulately interconnected for the drive rod 112 to follow the arc of the support . again , these are merely exemplary , not limiting , alternative embodiments in which one having ordinary skill in the art may incorporate the invention . addressing the drawings more specifically , the frame 114 may be a generally e - shaped member , having three flanges extending in the same general direction from a web . the two end flanges are drilled or bored or the like , to receive bearings or bushings or the like , to support the drive rod 112 . the middle flange is significantly thicker than the end flanges and is milled to receive the torque member 116 . the frame 114 is also provided with various mounting holes that may be required for use of the arm adjustment device , including to mount the brake 118 to the frame 114 . because the middle flange is thickened to receive the torque member 116 , brake mounting holes are conveniently located in the middle flange . although , one having ordinary skill in the art will realize that the brake 118 may also be mounted to either of the end flanges , and further that the brake 118 may be incorporated in the web portion of the frame 114 , all according to the desires of the user for any particular embodiment that incorporates the invention . further , one having ordinary skill in the art will note that the frame 114 may also be modified to be a generally u - shaped member , having two flanges extending in the same general direction from a web , and will also note that a u - shaped frame 114 is actually shown in the drawing fig1 and 2 . the torque member 116 is connected between the frame 114 and the drive rod 112 to &# 34 ; pre - load &# 34 ; the arm adjustment device and the arms 106 with a generally upward biasing force to counter balance the weight of the arms . the torque member 116 may be any of various known biasing devices , including a coil or helical spring , for example . most preferably , the torque member 116 is an elastic torsion spring . thus , the torque member 116 may comprise a hub and a perimeter collar with an elastic membrane extending between the hub and the collar and being bonded to each of the hub and the collar . alternatively , the elastic membrane may be a rubber bushing or the like that is force fit into the collar , with the hub being force fit into the bushing . either way , the elastic membrane or the rubber bushing may be tailored , with the use of commonly known methods and materials , to provide desired torsional spring characteristics for the invention . an off - the - shelf elastic torsion spring that is commonly available has been found to perform satisfactorily . to provide torsional force transfer between the frame 114 and the torque member 116 , the torque member may be keyed to the frame 114 by providing a slot and key in the frame aperture , with corresponding tab and slot being provided on the collar of the torque member 116 . to provide torsional force transfer between the hub and the drive rod 112 , the drive rod may be provided with a non - circular , preferably polygonal , cross - sectional shape , with the hub being provided with a corresponding hub aperture . as specifically shown in the drawings , the drive rod 112 is a hexagonal rod measuring about 7 / 16 of an inch across the flats . the drive rod support bearings are , therefor , hex bushings . also , the drive rod 112 may be milled or turned or the like with retainer seats 164 for holding the drive rod 112 in the frame 114 with c - shaped retainer clips 166 , or the like . the brake 118 comprises a brake rod and crank connection to the drive rod 112 and a jam clutch connection between the brake rod and the frame 114 . more specifically , the brake rod and crank connection comprises a pair of lever arms 170 , a drive pin 172 , a drive block 174 , and a brake rod 176 . each of the pair of lever arms 170 may be a generally rectangular or ovoid plate member that has opposing first and second ends . each lever arm 170 is provided with a hexagonal aperture 180 , near the first end , to mount the lever arms 170 on the drive rod 112 , with the lever arms 170 extending in the same general direction from the drive rod 112 . each of the two lever arms 170 is also provided with a pin aperture 182 , near the second end , to receive a bushing 184 to support the drive pin 172 . set screws 186 or the like may be used , as will be understood by one having ordinary skill in the art , to secure the lever arms 170 to the drive rod 112 and to the drive pin 172 as may be desirable . the drive pin 172 is a length of 3 / 8 inch steel rod that extends between the two lever arms 170 , and into the bushings 184 in the lever arms 170 . the drive pin 172 may have a set flat 190 to cooperate with a set screw 192 or the like of the drive block 174 . the drive block 174 may be an about 11 / 4 inch length of 3 / 4 inch square steel stock that is cross drilled ( see 194 ) to receive the drive pin 172 in sliding engagement . the drive block 174 may also be end drilled and tapped for the set screw 192 to secure the drive block 174 to the drive pin 172 . at an opposing end 196 , the drive block 174 is drilled and tapped to receive a threaded end 198 of the brake rod 176 . the brake rod 176 may be a 33 / 8 inch length of 7 / 16 inch square steel stock that has two opposing side surfaces textured ( fig3 ). a relatively short length ( about 5 / 8 inch ) of the brake rod 176 is threaded at one of two opposing ends 198 for screw engagement with the drilled and tapped aperture 196 of the drive block 174 . the brake jam clutch comprises a backing plate 200 , a top brake clutch 202 , a bottom brake clutch 204 , two brake springs 206 , and the brake actuator 120 . the backing plate 200 may be a generally rectangular plate that is about 11 / 2 inches wide , 31 / 8 inches long , and about 7 / 16 inches thick . the bottom brake clutch 204 is substantially an about 11 / 4 inch wide , 31 / 8 inch long , and 3 / 8 inch thick articulated member having a hinge plate 208 and a clutch plate 210 . the hinge plate 208 is also a generally rectangular member that is about 11 / 4 inches wide by 13 / 16 inches long , and 7 / 16 inch thick . a hinge tongue 212 extends about 7 / 16 of an inch out from one of the two long sides of the hinge plate 208 , in a direction generally away from the opposing long side of the hinge plate 208 . the tongue 212 is generally centered on the side of the hinge plate 208 , and is cut down to a width of about 3 / 4 inch and may be milled back about 1 / 16 of an inch from one face of the hinge plate 208 . the tongue 212 extends about 7 / 16 of an inch out from the side of the hinge plate 208 . the clutch plate 210 is an about 27 / 8 inch long plate having a width of about 11 / 4 inch and a thickness of about 3 / 8 inch . the clutch plate 210 has a cooperating notch 214 , corresponding to the tongue 212 of the hinge plate 208 . an about 1 / 2 inch by 9 / 16 inch jam hole 216 extends through the clutch plate 210 for the brake rod 176 to extend through the clutch plate 210 . the two short sides of the jam hole 216 are slightly sloped , or beveled , as shown ( fig2 & amp ; 27 ). the bottom brake clutch plate 210 also has an actuator cable aperture 218 and cooperating set screw 220 for clamping a brake actuator cable 222 . the top brake clutch 202 is substantially similar to the bottom brake clutch 204 , with the only notable distinction being a reverse of the slant for the two beveled sides of the brake rod jam hole 216 . the jam clutch is assembled with the backing plate 200 sandwiched between each of the upper and lower clutch plates 210 . the springs 206 or the like are interposed between the backing plate 200 and the clutch plates 210 , to hold the clutch plates spaced from the backing plate 200 , in an engaged position . the clutch springs 206 may be a helical coil or a leaf spring , for example . most preferably , the clutch springs 206 are resilient pad springs , that is pads of resilient , compressible material . in operation , the upper and lower clutch plates 210 will normally be held by the springs 206 to engage the brake rod 176 . with the clutch plates 210 engaging the brake rod 176 , the upper clutch plate 210 will resist and upward movement of the brake rod 176 , while the lower clutch plate 210 will resist a downward movement of the brake rod 176 . thus , in combination , the clutch plates 210 will preclude or resist either upward or downward movement of the brake rod 176 . activation of the brake actuator 120 will draw the two clutch plates 210 toward one another , against the clutch springs 206 , to release engagement of the clutch plates 210 from the brake rod 176 . so disengaged , the rod 176 is free to move upward and downward relative to the brake 118 and frame 114 . with the brake actuator 120 activated and the brake clutch plates 210 disengaged , the arms 106 may be adjusted upward and downward . when the arms 106 are adjusted downward , the drive rod 112 rotates in a first direction with the lever arms 170 rotating with the drive rod 112 to crank the drive pin 172 , drive block 174 , and brake rod 176 in a generally upward direction . conversely , when the arms 106 are adjusted upward , the lever arms 170 rotate with the drive rod 112 to move in an opposing , downward direction . the elastic torsion spring 116 applies a torsional force to the drive rod 112 to counterbalance the weight of the arms 106 . thus , a user may adjust the arms 106 with ease , not fighting the weight of the arms 106 themselves . it will be understood by one having ordinary skill in the art and by those who practice the invention , that various modifications and improvements may be made without departing from the spirit of the disclosed concept . various relational terms , including left , right , front , back , top , and bottom , for example , are used in the detailed description of the invention and in the claims only to convey relative positioning of various elements of the claimed invention . the scope of protection afforded is to be determined by the claims and by the breadth of interpretation allowed by law .

an adjustable chair arm is used with a chair that has a base , a seat supported by the base , and a back connected with the base . the arm is an elongated member that is connected with the base and has two opposing ends . an arm adjustment device is interposed between the arm and the base and has a drive rod , a frame , a torque member , a brake , and a brake actuator . the frame holds the drive rod in rotating engagement . the torque member is connected between the frame and the drive rod and applies a torsional force to the drive rod . the brake has a locked mode and a released mode and holds the drive rod in preselected rotational positions , relative to the frame . the brake actuator manipulates the brake between the locked and released modes .

referring now to the drawings , and particularly to fig1 thereof , a typical set of components of a grill that may be packaged in a shipping container in accordance with the present invention are shown . the components of the grill include a firebox assembly 10 , a pair of side tables 12 and 14 , and a cart assembly comprising a front and rear support assemblies 18 and 20 , respectively , a pair of cross - members 22 , a pair of feet 24 , a pair of wheels 26 , and a pair of hitch pins 28 for mounting the wheels . the firebox assembly 10 comprises a firebox base 30 for the valves , burners and grills ( not shown ) typically employed for applying heat to the food . as shown , the firebox base has burner control knobs 32 on a front panel 34 and a base support 36 for supporting it on top of the cart assembly at approximately the waist level of a typical adult , and a firebox hood 38 with a handle 40 for selectively covering and uncovering the firebox base . the firebox hood is appropriately hinged to the firebox base ( not shown ) to allow the hood to be opened and closed , as is well known in the art . one of the side tables 14 also has a valve , burner and grill or the like ( not shown ), covered by a hinged lid 42 , with a control knob 44 ( fig6 ) at the front . the firebox assembly also comes with a hose assembly 46 for connecting a supply tank ( not shown ) to the valves and burners in the firebox base and to the valve and burner in the side table . the front support assembly 18 comprises a pair of legs 48 held in spaced - apart relationship by a front panel 50 , and the rear assembly comprise a similar pair of legs 52 held in spaced - apart relationship by upper and lower cross - members 54 and 56 , respectively . the front panel serves both an aesthetic purpose and to add structural rigidity to the front support assembly . preferably all of the legs are tubular , and the legs , the front panel and the upper and lower cross - members are made of steel or other suitable structural material . the front panel is fastened by threaded fasteners to the legs of the front support assembly , and the upper and lower cross - members are spot - welded to the legs of the rear support assembly , though the front panel and cross - members can be attached in any other suitable manner . as seen in fig1 , the support assemblies 18 and 20 are packaged in the shipping carton folded together so as to be relatively flat and compact . to this end , as shown in fig2 a and 2b , the two support assemblies are partially pre - assembled by interconnecting them with a pair of linking arms 58 that extend between opposing pairs of legs on the support assemblies . a hinge 60 in the mid - portion of each linking arm divides it into first and second linking arm portions 58 a and 58 b . the opposite ends of each linking arm are pivotally connected by pins or screws 62 to the inside surface of a leg . as a result , the linking arms are capable of folding onto themselves . the pins of the hinges are located at the top of the linking arms so that the linking arms fold downward , though they could be designed to fold upward or inward . when completely folded , each linking arm is received along the insides of the opposing legs to which it is pivotally connected , allowing the two support assemblies to collapse together with little or no separation between them , thus minimizing space in the shipping carton . when the support assemblies 18 and 20 are removed from the shipping carton , they can be unfolded by the assembler as shown in fig2 a . when completely unfolded as shown in fig2 b , the linking arms 58 will position the support assemblies in approximately the proper position for further assembly of the grill . in this regard , it should be noted that the linking arms in this first embodiment of the invention are not intended to be permanent structural elements of the completely assembled grill . rather , the linking arms are only intended to quickly and properly position the two support assemblies and hold them sufficiently steady in the front - rear direction so that an assembler , working alone , can readily attach other components without need of an assistant . therefore , the linking arms can be made of less expensive material such as plastic . to help ensure that the linking arms 58 hold the support assemblies in the proper position while the assembler attaches other components , each linking arm is provided with a locking mechanism at or near its hinge 60 . as shown in fig3 a - 3c , the locking mechanism comprises a cylindrical protrusion 64 formed in the end of one linking arm portion 58 a that mates with a complementary recess 66 formed in the abutting end of the other linking arm portion 58 b . the protrusion and the recess are sized to mate with an interference fit to lock the two linking arm portions together , though persons of ordinary skill will appreciate that other locking schemes could be used . alternatively , or in addition , suitable locking mechanisms could be employed where the linking arms pivotally connect to the legs of the support assemblies . where the linking arms are not intended to serve as permanent structural members , the locking mechanisms need not be very robust , but only sufficient to hold the support assemblies steady while the assembler attaches other , permanent structural components such as the cross - members 22 . once the support assemblies 18 and 20 are completely unfolded , the assembler can attach the permanent cross - members 22 to opposing legs 48 and 52 of the two support assemblies to permanently lock them in the proper position . in order to make this process quicker and easier , as shown in fig2 b the support assemblies are shipped with threaded fasteners 68 for attaching the cross - members that have been partially threaded in place in threaded holes formed in the legs . these threaded fasteners are preferably held in place with a common thread locking compound , such as loctite ®, to prevent the fasteners from falling out due to vibration during transit , while still allowing the fasteners to be tightened during grill assembly . in addition , vertically - oriented keyholes 70 are formed in the ends of the cross - members to mate with the fasteners . the larger openings of the keyholes are sized to be received over the heads of the threaded fasteners without having to remove them , and the smaller openings of the keyholes are sized to slide around the threaded shafts of the fasteners , underneath the fastener heads . once the keyholes in the cross - members are placed over the fastener heads and slid into place , the fasteners can be tightened to secure the cross - members to the legs as shown in the enlarged fragmentary view in fig2 b . with the cross - members 22 secured in place , the assembler can then install the feet 24 on the legs on one side of the support assemblies 18 and 20 , and the wheels 26 using the hitch pins 28 inserted through holes on the legs on the other side the support assemblies ( fig4 ). still referring to fig4 , the firebox assembly can now be installed on the cart assembly . to this end , a connector block 72 is sized and shaped to fit into the end of each of the legs of the support assemblies , with a portion of each connector block protruding out of the end of the legs . the base support 36 of the firebox assembly 10 has four downward extending tubular stub legs 74 that align with the legs of the support assemblies . a front stub panel 76 extends between the tubular stub legs of the base support . the protruding ends of the connector blocks are sized and shaped to be received in open ends of the tubular stub legs of the base support . the connector blocks can be secured to the legs of the support assemblies and to the stub legs of the base support by threaded fasteners inserted through the walls of the legs and the stubs into the connector blocks ( not shown ). in addition , or in the alternative , the firebox assembly can be secured to the cart assembly by brackets 78 held in place by threaded fasteners received in slotted openings 80 in the brackets , such as shown in the enlarged illustration in fig4 . fig5 illustrates attachment of one of the side tables 12 to the side of the fire box assembly 10 . preferably the firebox base 30 is shipped with threaded fasteners partially threaded into holes for attaching the side tables . the side table includes keyhole openings 70 that are sized and positioned to receive the fasteners . to install the side table , the assembler aligns each keyhole with its respective fastener so that the heads of the fasteners are received through the larger openings of the keyholes . the assembler then slides the side table so that the smaller openings of the keyholes are received underneath the fastener heads and then tightens the fasteners . the other side table 14 may be installed in a similar manner . the completely assembled grill is shown in fig6 . as noted , one of the side tables may include a side burner assembly ( not shown ). fig6 a illustrates attachment of a valve 84 for the side burner assembly to the side table . preferably the valve is shipped with threaded fasteners partially threaded into holes for attaching the valve to the side table . the side table includes an opening to receive the valve stem 86 of the valve and keyhole openings 70 that are sized and positioned to receive the listeners that have been pre - inserted in the valve . to install the valve , the assembler passes the valve stem and the fasteners through their respective openings in the side table so that the heads of the fasteners are received through the larger openings of the keyholes . the assembler then slides the valve so that fastener heads are received over the smaller openings of the keyholes and tightens the fasteners . once the side burner valve is in place , the assembler can install the burner control knob 44 onto the valve stem , as shown in fig6 b . an alternative embodiment of the support assemblies for the cart is shown in fig7 a and 7b . in this embodiment , the front and rear support assemblies 18 ′ and 20 ′, respectively , each again comprise a pair of legs 48 ′ and 52 ′, respectively , held in spaced apart relationship by upper and lower cross - members 54 ′ and 56 ′, and the two support assemblies are likewise partially pre - assembled by interconnecting them with a pair of linking arms 58 ′ that extend between opposing pairs of legs on the support assemblies . similarly , a hinge 60 ′ in the mid - portion of each linking arm divides it into first and second linking arm portions 58 a ′ and 58 b ′, and the opposite ends of each linking arm are pivotally connected to opposing legs of the support assemblies . the primary difference is that the linking arms are hinged to fold inward , rather than downward as in the embodiment shown in fig2 a and 2b . as in fig2 a and 2b , these linking arms in this embodiment are not intended to be permanent structural elements of the completely assembled grill , but rather are only intended to position the two support assemblies and hold them sufficiently steady while the assembler attaches other components . therefore the linking arms can be formed of less expensive material such as plastic , as in fig2 a and 2b . castors ( not shown ) can be installed in the bottom of each leg . with reference to fig8 a - 8c , each linking arm 58 ′ further includes a locking mechanism near the hinge 60 ′ similar to the locking mechanism shown in fig3 a - 3c . in this case , the locking mechanism is formed on the rear side of the linking arms and the two linking arm portions 58 a ′ and 58 b ′ are designed to overlap at the hinge for great structural integrity . in this regard , at the hinge , the end of one linking arm portion 58 a ′ is formed with a rectangular extension 88 that is received with a relatively tight fit in a complementary rectangular recess 90 formed in the other linking arm portion 58 b ′. as a result of this overlap , the linking arm is more rigid in the vertical direction when completely unfolded and locked . in addition , a cylindrical protrusion 64 ′ is formed on the end of the rectangular extension that mates with a complementary recess 66 ′ formed in the end wall 92 of the rectangular recess . similar to fig3 a - 3c , the protrusion and the recess are sized to mate with an interference fit to effect locking . referring again to fig7 a , a bottom shelf 94 is pivotally connected as by a hinge or the like along its rear edge to the lower cross - member ( not shown ) of the rear support assembly 20 ′. this allows the bottom shelf to be folded into the rear support assembly for compactness when the two support assemblies 18 ′ and 20 ′ are folded together for shipping . after the support assemblies are unfolded and the linking arms 58 ′ are locked in position , the assembler can fold the bottom shelf down into position and secure it by fasteners ( not shown ) to the lower cross - member 56 ′ or to the legs 50 ′ of the front support assembly . as shown , the bottom shelf includes a relatively large circular opening 96 for holding a supply tank . an alternative embodiment of the folding shelf design of fig7 a - 7b is shown in fig9 a - 9c . in this embodiment , the front support assembly 18 ″ has legs 48 ″ held in spaced - apart relationship by upper and lower cross - members 54 ″ and 56 ″, which can be spot - welded in place , and the rear support assembly 20 ″ has legs 52 ″ held in spaced - apart relationship by a rear louvered panel 98 that has been pre - inserted with fasteners ( not shown ). the two support assemblies are interconnected by linking arms 58 ″, each of which has a hinge 60 ″ in its mid - portion dividing it into linking arm portions 58 a ″ and 58 h ″, and the opposite ends of each linking arm portion are pivotally connected to opposing legs of the support assemblies . each linking arm includes a locking mechanism near the hinge similar to fig3 a - 3c and fig8 a - 8c . a bottom shelf 94 ″ with a relatively large circular opening 96 ″ for a supply tank is pivotally connected to the rear panel as by hinges ( not shown ), and visible through the opening are louvered side panels 100 with hand openings 102 that are pivotally connected to each side edge of the bottom shelf by hinges 104 ( fig9 b ). as in the embodiment of fig7 a - 7b , this allows the bottom shelf with its two side panels to be folded substantially into the rear support assembly for compactness when the two support assemblies are folded together for shipping . after the support assemblies are unfolded and the linking arms are locked in position , the assembler can fold the bottom shelf down into position and secure it by fasteners ( not shown ) to the lower cross - member or the legs of the front support assembly . the side panels can then be pivoted into position between opposing legs of the front and rear support assemblies and fastened in place as by fasteners 68 . castors 106 are installed in the bottoms of the legs , and as shown in fig9 c , a pair of hinged doors 108 with handles 110 can be installed on brackets 112 on the front to complete the cart assembly . a firebox assembly ( not shown ) can then be mounted on top of the cart assembly . an alternative embodiment of a linking arm 58 ′″ with a locking mechanism that can serve as a permanent structural support for a cart assembly is shown in fig1 a - 10d . as with the other linking arms described previously , a hinge 60 ′″ in the mid - portion of the linking arm divides it into first and second linking arm portions 58 a ′″ and 58 b ′″. in this case , each linking arm portion is formed as a channel with end walls from steel or other suitable structural material . the two linking arms portions are pivotally connected at their outer end walls to opposing legs of two support assemblies ( not shown ) and the inner end walls of the two linking arm portions abut at the hinge 60 ′″. a locking arm 114 in the form of an angle member , also formed of steel or other suitable structural material , is permanently attached to the upper surface of the first linking arm portion 58 a ′″, as by spot welding , so that the angle member extends beyond the inner end wall of the first linking arm portion and overlaps the second linking arm portion 58 b ′″ when the linking arm is fully unfolded . the second linking arm portion 58 b ′″ has a slidable locking pin 116 mounted in a u - shaped bracket 118 that is permanently attached , as by spot welding , to the inside wall of the second linking arm portion . a bias spring 120 is sandwiched between the top of the u - shaped bracket and a collar on the locking pin ( not shown ), biasing the upper end of the locking pin so that it extends through a hole 122 formed in the upper surface of the second linking arm portion . a like hole 124 is formed in the upper surface of the locking arm on the first linking arm extension , positioned to receive the locking pin when the linking arm is fully unfolded . the leading edge of the locking arm is raised where it engages the locking pin to allow it to pass underneath the locking arm and enter the hole . in this manner , the first and second linking arm portions can be securely locked together in the fully unfolded state so that they may serve as structural support members of the cart assembly and avoid the use of additional structural members in lieu of the linking arm . it will be appreciated that the various embodiments of the invention described above provide a grill having a cart assembly that can be partially pre - assembled using one or more linking arms , shipped in a folded configuration , and then quickly unfolded and held in the proper position while an assembler , without anyone &# 39 ; s assistance , can assemble the cart assembly and mount a firebox assembly to it . assembly is further simplified by having fasteners partially threaded in place and forming keyholes or slotted openings in components to be attached by those fasteners . the invention thus allows the grill to be shipped compactly yet more quickly and easily assembled at its destination , saving time and labor costs . the present invention has been described above in terms of presently preferred embodiments so that an understanding of the present invention can be conveyed . however , there are other embodiments not specifically described herein for which the present invention is applicable . therefore , the present invention should not to be seen as limited to the forms shown , which are illustrative rather than restrictive .

the present invention is embodied in a grill configured for quick assembly , a method of partially pre - assembling the grill for shipping from the factory in a container , and a method of assembling the grill when removed from the shipping container . the grill comprises a firebox assembly for mounting on a cart assembly , the cart assembly having first and second support assemblies and at least one linking member interconnecting them . a hinge or pivot in the mid - portion of the linking member divides it into first and second linking member portions , and the opposite ends of the linking member portions are pivotally connected by hinges or the like to the first and second support assemblies . the linking member holds the first and second support assemblies together in a partially pre - assembled state , and the pivotal connections of the linking member portions allow the two assemblies to be folded for packaging in a shipping container . when removed from the container , the pivotal connections of the linking member portions allow the first and second support assemblies to be unfolded , with the linking member holding the two support assemblies in approximately the proper position with respect to each other and to the firebox assembly for assembly of the grill , thus allowing for easier and quicker grill assembly by one person .

the simplest embodiment of the invention is a modification of prior art for laser lithotripsy as depicted in fig2 , the deficiencies of which are related to the susceptibility of the fiber tip 95 , particularly for smaller core fibers . for orientation , a silica core , fluorine - doped silica clad 85 , fluoroacrylate 80 coated and etfe 75 ( ethylene tetrafluoroethylene copolymer ) buffered optical fiber 70 is housed within a laser connector 60 and laser energy is provided to the fiber at 55 . a rubber boot 65 and / or layered heat shrink tubing provides bend limitation at the back of the connector 60 . laser energy is emitted from the tip in a frustoconical volume 90 where the divergence half angle is approximately 13 degrees ( arcsine of the fiber numerical aperture ). this latter parameter , the divergence angle , is somewhat controversial with some claiming the angle is lower due to the higher refractive index of the working environment : saline . this is true for wavelengths of laser energy that do not interact significantly with saline , but for the wavelengths most commonly used for laser lithotripsy , a steam bubble is formed between the fiber output face 95 and the target where the steam has a refractive index that is essentially the same as that of air . with the fiber operating in steam , the divergence is essentially the same as it is operating in air . small core fiber tips degrade rapidly in use when addressing calculi in a process referred to as “ burn back ” in the interventional urology art . burn back rates appear to be proportional to pulse energies in holmium laser lithotripsy but this observation may be an artifact ; initial damage to the fiber output face may be independent of pulse energy where the consequences to the damaged fiber tip are more dramatic at higher pulse energies as more energy is back - scattered . stone ablation rates suffer greatly after onset of fiber tip damage . the source of tip damage initiation has not been determined but it is highly probable that the onset is similar in cause to damage to other types of fibers that have been better characterized : side fire fibers . in side fire fibers , transmissive surface damage initiates due to tissue adhesion about the output on the protective cap . where the energy density of the distorted semi - gaussian beam profile is insufficient for vaporizing tissue , tissue adheres , forming a ring about the higher irradiance center of the output spot . this ring of tissue cooks , then carbonizes , absorbing the sub - therapeutic laser energy in the beam periphery more strongly as it blackens and transferring that heat to the protective cap . fused silica protective caps have low thermal conductivity such that the conducted heat is highly localized and temperatures about the output spot raise enough to lower the viscosity of the amorphous silica such that , in combination with intercalating alkali metal ions and counterions , water , etc ., the silica rearranges structurally to energetically favorable crystalline form : high cristobalite ; this is devitrification . crystalline silica is birefringent so the optical transmission characteristics subtly change , but the more likely problematic change to the cap is crystallite formations that scatter the laser energy , may slough off , etc . distortion the output beam profile further and further , amplifying the progression of devitrification , ad infinitum until a pit if formed in the protective cap at the output . as discussed in the background section , aqueous irrigant used in urological surgery absorbs roughly 40 % of infrared laser radiation at as little as 1 mm separation between the fiber and the target . contact with tissue or calculi initiates damage to the fiber and reduces efficient coupling of laser energy to the target by roughly 40 % within minutes . it is impossible to precisely control fiber to target distance with endoscopic visualization and manual control . accordingly , surgeons place the fiber tip in direct contact with the calculus to minimize the energy lost to boiling water . while calculi are not organic tissue , per se , they often contain organic crystals and , when primarily inorganic , stones often harbor considerable organic waste in boundaries between crystals so a similar damage onset cause is probable . even absent carbonization , direct contact between the fiber tip and material under laser ablation causes the local temperature increase necessary for devitrification onset . once the tip is damaged the conditions for accelerating devitrification improve and provide the positive feedback loop described for side fire fiber pitting discussed above . fig3 depicts a simple embodiment of a solution to this problem : a ceramic , glass , polymer or metal ferrule 105 is mounted on the fiber tip at the fluoroacrylate coating 110 with adhesive 115 . the ferrule 105 serves two functions : establishing a fixed and short stand - off from the target and excluding saline infusion into the optical path 135 . the embodiment illustrated in fig3 provides as a simple retrofit of the standard laser lithotripsy fiber depicted in fig2 . in fig2 , the bare glass clad 85 portion of the fiber tip is illustrated as it appears after test firing in air , if somewhat exaggerated in length , in anticipation of the consequent bare fiber section 125 thought probable to result from test firing the new art in air after addition of the ferrule 105 . ( 100 % of surgical fibers are functionally tested on surgical lasers prior to sterilization as part of the quality system at innovaquartz llc .) the ferrule includes a borehole that is preferably a longitudinal , centrosymmetric opening in the tube / ferrule . as used herein , the borehole can include a plurality of sections or regions . a single section may be referred to a bore , whereas a plurality of sections / regions can be distinguished based on the inside diameter ( id ) of the borehole and are referred to as a first bore , a second bore , and / or a third bore , or other distinction that clearly indicates the differences between the sections / regions based on id . while adhesion to the etfe buffer 100 of the fiber is difficult , adhesion to the fluoroacrylate fiber coating 110 is not a great deal better and , being opaque , a ceramic , metal or polymer ferrule 105 makes inspection of the coverage of the adhesive 115 in the bond impossible . further , for adherence to basic engineering principles , at least two methods of retaining the ferrule on the fiber is preferred , particularly where the consequences of ferrule detachment are problematic ; a considerable surgical time would be spent in attempting to retrieve a lost ferrule from minor renal calices where it could seed formation of additional kidney stones . small fibers are used in laser lithotripsy because larger fibers are incompatible with the flexible ureteroscopes needed to reach stones in the kidney , particularly the ‘ lower pole stones ’ that gravity favors . working channels are typically 3 . 6 fr or just over a millimeter in diameter . while it is possible to pass a 365 μm core fiber through a 3 . 6 fr channel , the 0 . 8 mm to 1 mm etfe jacketed fiber blocks irrigation flow almost entirely and the stiffness of the 0 . 4 mm glass fiber ( 1 . 1 cladding to core ratio ) restricts the deflection of the scope to near inutility . 273 μm core fibers ( generally known as 272 μm core , due to an early rounding error ) are the largest that are commonly used with flexible ureteroscopes when addressing renal calculi . the typically 0 . 45 mm etfe outer diameter allows passage of sufficient irrigation for maintaining clarity in the surgical field but the 0 . 3 mm glass fiber does restrict scope deflection sufficiently to make accessing lower pole stones extremely difficult with the fiber preloaded in the working channel . boston scientific &# 39 ; s accutrac ™ and flexiva ™ tractip 242 μm core fibers are designed to pass through the working channel while it is fully deflected , eliminating fiber rigidity related reduction in scope deflection , at least for initial placement of the fiber ( the fibers are designed to pass the deflected working channel one time , only ). the slightly up - tapered and lens - ended fiber tip of the boston fibers also reduce divergence in the area immediately adjacent to the fiber output and the larger emission diameter , and likely the absence of sharp edges , forestalls burn back with less than 2 mm loss of length being reported for typical lithotripsy cases . in reality , however , the boston scientific fibers ( 1 . 2 ccdr , or cladding to core diameter ratio ) are little more flexible than a standard 273 μm core fiber ( 1 . 1 ccdr ) and recent reports from the field indicate that burn back continues to be a major concern in ureteroscopic lithotripsy ( urs ). the area immediately adjacent to the fiber emission face is the area where most energy is lost to the moses effect . it is impossible to hold a fiber perfectly flat against an irregular kidney stone surface such that at least some portion of the energy must vaporize water in transit to the stone surface , and the fraction of the laser energy lost increases proportionally with pitting of the calculus by ablation and geometrically with fiber output face damage . 200 μm core fibers , such as depicted in fig2 and 3 , are the most commonly used fibers for accessing lower pole kidney stones because they permit full deflection of all scopes when preloaded within the working channel . burn back in 200 μm is a considerable problem and the damage typically begins within the first 2 minutes of lasing . when the fiber burns back to the etfe buffer , many surgeons will consider removing the fiber and for intraoperative reprocessing : strip the etfe jacket and cut a new fiber output face . smaller core fibers , e . g . 150 μm core , are even more susceptible to burn back , and larger core fibers , e . g . 273 μm core , are also susceptible to burn back , although somewhat less so . the techniques used to perform this reprocessing are generally crude and far from optimal , generally involving cutting the fiber with scissors . nicks to the fiber are commonly produced due to the use of inappropriately sized fiber strippers , damaged stripper blades ( high carbon steel blades start to rust even before the first use , due to autoclave sterilization ) or an almost ubiquitous use of improper technique . tips detaching on reintroduction through even a somewhat relaxed working channel is a common complaint . passing a working channel with a sharply edged , freshly cut fiber under any deflection risks the tip penetrating the working channel liner ( typically a thin walled fluoropolymer ) or at least generating pits and scratches that harbor waste and bacteria inaccessible to disinfection and passing these materials on to the next patient ( ureteroscopes cannot be sterilized ). ( efforts at educating surgical personnel in proper fiber reprocessing techniques have largely failed .) patients under general anesthesia suffer higher risk with time under anesthesia such that time in the or should be kept to a minimum . or time also costs a great deal of money and is typically billed by the minute . the extension of a surgical session due to the almost immediate loss of optimum fiber to target coupling efficiency , removing and reprocessing fiber tips or simply removing and reloading fibers , repositioning the ureteroscope , chasing after detached fiber tips , etc . generally exceeds the total lase time . the fiber depicted in fig3 also slightly focuses the laser energy 140 due to a curved output face 130 , enabling a close fitting ferrule 105 to be used without blocking the higher angle rays of the otherwise immediately diverging output . air is trapped in the hollow space produced by the fiber setback 145 within the ferrule and this bubble is maintained throughout insertion of the fiber into the saline - filled surgical field due to the very strong barrier that the surface tension creates at such small dimensions . setback 145 is illustrated at approximately the maximum attainable without spatially filtering or ‘ clipping ’ the emitted beam 140 of the 200 μm fiber depicted and to provide a reservoir for air . in use , the ferrule 105 of the fiber is maintained in contact with the target so that it gets hot , albeit not as hot as the much smaller , bare fiber where ceramic , polymer or glass ferrules are employed . metal ferrules may get hot enough to melt the fiber buffer material 100 and / or damage the adhesive bond , promoting tip detachment . under any heating , the air bubble captured in the open bore of the ferrule 135 expands and some air may be lost as a result . when the fiber is at rest , cooling , the smaller amount of air occupies a smaller volume within the open bore 135 , generally predicted by using charles &# 39 ; law to estimate the gas expansion assuming the expanded gas is all lost : v1 is the volume of the open bore , t1 is physiological temperature in kelvins , v2 is the expanded volume t2 is the maximum temperature reached . the result of this simple exercise is less than 25 % of the gas bubble is lost for every 100 degree ( centigrade ) temperature rise , independent of the initial volume , and less than 20 % total , if one assumes cooling by the surrounding irrigation flow keeps the temperature at or under 100 ° c . where fluid does intrude into the bore of the ferrule during periods of rest , it is displaced upon the first pulse on next activation but it will return between pulses ( unless the pulse rate is faster than the collapse of the moses bubble — rates unachievable using existing holmium lasers ) but will not return where cw lasers are used until cessation of lasing . the mass of water that must be vaporized with each pulse in a second ( and subsequent ) treatment session remains constant baring condensation of droplets deeper within a hydrophilic ferrule bore , therefore a hydrophobic interior surface is desirable . the mass of water vaporized is also directly proportional to the depth of the setback 145 , so minimization of setback is also desirable . fig4 illustrates a preferred embodiment of a fiber tip equipped with a fused quartz ferrule 150 with minimal setback 185 , at approximately 0 . 25 mm , where post - production functional testing was performed in water rather than air such that the burn back 180 of the hydrophobic fluoroacrylate ( fa ) coating 175 is minimized and due solely to the formation of the slight convex lens 190 on the fiber tip 215 and the fresnel reflections thereby produced . fig4 a depicts the air bubble 200 slightly protruding 195 prior to first laser activation and fig4 b depicts the air bubble 220 with approximately 15 % of the volume replaced by water 210 prior to a subsequent laser treatment session ( and following reaching about 100 ° c . in the first session ). features of the device are a hydrophobic and rounded surface 205 where the ferrule 150 contacts the kidney stone , a hydrophobic surface within the setback volume 200 and 220 , a rounded proximal end 225 to facilitate passage through scope channels , two different diameter bores , the larger 155 of which accommodates the nylon buffer 170 , preferred for much greater adhesion with uv cure adhesives such those offered by dymax , norland and electronic materials and the smaller 160 of which accommodates the fa coated fiber 175 . assembly is simplified by the dual diameter bore and transparent ferrule . the bare fa fiber segment is threaded into the silica ferrule and is advanced past the chamfer 165 between the bore diameters . just prior to the nylon entering the large bore 155 , an annulus of adhesive is applied about the bare fa fiber just distal to the nylon buffer 170 . as the fiber is advanced to a point where it stops , with the nylon on the chamfer 165 , excess adhesive is forced into the thin cylindrical spaces between the bare fa coated fiber and the nylon buffered fiber . inspection to verify sufficient adhesive coverage is easily performed via microscopic examination through the transparent silica ferrule ( sleeve or sheath ). uv light cures the adhesive through the uv transparent ferrule . the rounded stone contact or distal end of the fiber ferrule 205 serves several purposes . it facilitates passage through even fully deflected ureteroscopes , it alters the contact angle of the meniscus at the air water barrier , supporting a protrusion of air 195 that is small enough to be sustained , even in rough handling , and it minimizes the contact and thermal conduction between the fiber assembly and the target . partial round ends are more robust than full round ends but both options have utility when the ferrules are made of fused quartz , fused silica , borosilicate glass and alternative materials such as sapphire , zirconia , alumina and other ceramics . metallic ferrules offer a further potential for retention on the fiber by crimping onto the fiber buffer at or about 170 but metals quite efficiently conduct heat to heat labile portions of the fiber assembly and may be of limited utility in practice . fig5 depicts a preferred embodiment directed to replenishing the air bubble 225 should water find its way to the ‘ bottom of the air well ’— where the end of the fiber 230 defines the depth 235 of the air bubble 225 . the ferrule ( or sleeve ) in this embodiment is ½ rounded or filleted 240 at the stone contact face 245 to permit smooth passage through the working channel under full deflection while presenting a more robust interface with the stone than that depicted in fig4 . the rear or fiber insertion end 250 of the ferrule is chamfered 255 on the outer diameter to reduce the surface area of contact with the working channel liner and chamfered 255 on the inner diameter to facilitate threading the delicate , bare fiber 260 into the ferrule and for loading of a moisture - sensitive , gas - generating charge 265 ( e . g ., a stoichiometric mixed bed of citric acid and sodium bicarbonate or other biocompatible gas generating compounds , e . g ., those described in u . s . pat . no . 3 , 556 , 803 and incorporated herein by reference in its entirety ) within a space provided by an intermediate bore diameter 270 . the proximal end of the ferrule is also rounded 290 to provide for smooth passage of the fiber in removal . the fiber fa coating 275 has been removed from the distal portion of the fiber 260 , terminating 280 just distal to the edge 285 of etfe or nylon buffered fiber . adhesive fills the void 310 between the buffered fiber 305 and the large bore 295 of the ferrule and part of the void 311 between the intermediate bore diameter 270 and the fa coated 275 and bare fiber 260 , up to the mixed bed of gas generating compounds 265 . in use , where saline intrudes to the bottom of the well at the outer circumference 300 of the fiber tip , it wicks between the bare fiber 260 and the small bore 315 of the ferrule , by capillary action , until it encounters the gas generating bed 265 . upon contact with liquid water , the gas generation bed produces gas ( co 2 in the case of the sodium bicarbonate citric acid mix ). the gas generated displaces the water and refills the air bubble well 225 . for laser lithotripsy , the local heat generated in ablating the kidney stone — this invention is primarily directed to kidney stones and gall stones as opposed to ureter stones or bladder stones although the embodiments taught herein , and larger scale embodiments thereof , may find applications in other surgeries such as soft tissue ablation — has proven sufficient to initiate burn back wen confined to small and bare fiber tips . thermal issues are typically far more problematic for soft tissue surgery application of optical fiber , particularly where the fiber may be encrusted with carbonized tissue or become inadvertently buried in tissue ( or purposefully so ). for higher temperature uses like soft tissue ablation , ceramic , sapphire or other refractory material ferrules are clearly superior to the fused quartz or fused silica and alternative geometries may prove superior . sapphire ferrules &# 39 ; transparency , lack of susceptibility to devitrification and refractory qualities may also offer simplified and reliable assembly and superior longevity when used on the small fibers typically used urs . for kidney stone applications with access via flexible ureteroscope , working channel space limitations preclude the addition of coaxial fluid communicating conduits in most cases , particularly where significant flows are necessary for the function . an embodiment of the invention that remains dimensionally compatible with the just over 1 millimeter lumen of the working channel is illustrated in fig6 . some flexible ureteroscopes have working channels smaller than 1 mm , e . g . a dual 3 . 3 fr channel scope from richard wolf , within which even the embodiment depicted in fig6 , with an outer diameter of 0 . 65 mm could restrict the flow of irrigation too much . the fluidic delivery embodiment utilizes a ceramic ferrule 340 because materials such as alumina are less likely to fracture with the thin walls that are required for space considerations ; in this case , for reference , the bore of the ceramic ferrule is just slightly larger than the fa coating 345 diameter of the standard 200 μm core fiber ( 240 μm glass clad , 260 μm fa coating 345 , 400 μm etfe buffer 330 ) with an outer diameter 350 of 0 . 55 mm . the fiber in this instance is shown as mechanically flat polished at the output face 360 and the inherently hydrophobic fa coating 345 remains intact . flat polished fibers of the type used in holmium laser lithotripsy are 0 . 22 ± 0 . 02 na ( numerical aperture ). the divergence of these fibers is therefore approximately 0 . 22 radians so that a flat polished fiber emitting energy at maximum divergence cannot be set back within a closely matching bore any appreciable distance 385 without some of the energy contacting the bore wall . in illustrating the fluidic delivery embodiment ( fig6 ), the setback 385 depicted is approximately the maximum that it could be give the divergence 380 of the output , were the outlet end of the ceramic ferrule not chamfered 370 , but in order to avoid chamfering the output end of the ferrule , the fiber centricity within the fa coating 345 and fa coating centricity within the bore of the ferrule would have to be very close to perfect , as would the setback distance , to avoid losing any energy at all in absorption by the ferrule 340 . in that one of the fundamental goals of the invention is to minimize losing precious laser energy in heating not target materials , particularly saline , chamfering the ferrule is a reasonable precaution where flat output fibers are used . the ceramic ferrule is ½ rounded ( filleted ) 390 on each end . the fluid transfer tube 320 is a multilumen and thin wall structure of polymer such as polyimide , e . g . hd microsystems pyralin p12542 , a fluoropolymer such as fep , or other polymer with a large central lumen 325 slightly larger than the etfe buffer 330 diameter of the fiber , surrounded by a plurality of small lumen 365 . the distal end of the multilumen tube 320 is counterbored to accommodate the ceramic ferrule diameter and the proximal end of the ferrule 340 seats in the multilumen tube 325 where the etfe buffer abuts 335 the ceramic ferrule 340 . fluid is passed thorough the plurality of small lumen 365 about the circumference of the central lumen 325 , communicating with the multilumen tube by way of a t or y fitting , located between approximately 50 cm from the distal terminus and proximal terminus of the device , as known in the art . the fluid delivery lumens are partially obstructed by the ceramic ferrule within the counterbored multilumen tube , producing a localized pressure rise that is a function of the fluidic supply parameters and fluid viscosity . the purpose of providing the fluid to the laser emission end of the device is to displace the saline irrigation just about the fiber and preferably substantially between the fiber and the target , with an infrared transparent , non - toxic , biocompatible material , preferably of low viscosity and with a density greater than water / irrigant saline . potential candidate liquids meeting these requirements are the fluorinert electronic heat transfer fluids made by minnesota mining and manufacturing . alternatively , it may be desirable to utilize a coaxial fluid conduit such as depicted in fig6 in the reverse : suction applied to the t or y fitting to remove stone dust and debris from the surgical field . the preceding embodiments can include an open or exposed fiber output face ( e . g ., 130 in fig3 and 190 in fig4 ) where an air bubble or the moses bubble prevents the interaction of the saline and / or biological fluids with the fiber output face . preferably , biological fluids do not contact the fiber output face . to improve the retention of the air bubble or moses bubble at the fiber output face , the ferrule can include a hydrophobic or super - hydrophobic surface . the hydrophobic or super - hydrophobic surface is preferably located on the external output surface and within the bore , preferably , covering the internal setback surface , in one instance , the hydrophobic surface is includes within the borehole ( preferably covering the setback surface ). the hydrophobic or super - hydrophobic surface can , for example , include a fluorosilane ( e . g ., the condensation product of an organosilane ( r f — si ( or ′) 3 ) with a silica surface ), a nanoparticle surface ( e . g ., a nanotexture film ), or a patterned surface ( e . g . a nanotexture ) whereby the coating or surface structure provides the desired hydrophobicity . in one instance , the desired hydrophobicity can be expressed in terms of a saline contact angle , the contact angle is preferable greater than 90 °, 100 °, 110 °, 120 °, 130 °, or 140 °. as described above , the utilization of the laser can heat the quantity of gas within the setback volume ( e . g ., 200 and 220 in fig4 ) which may decrease during firing ( bubble loss ); upon cooling this decrease in quantity ( temperature and pressure ) can lead to water entering the setback volume ( compare fig4 a and fig4 b ). preferably , the hydrophobic or super - hydrophobic setback surface provides a contact angle of at least 90 °, 100 °, 110 °, or 120 ° even when the pressure within the setback volume is decreased due to quantity loss . even more preferably , the hydrophobic or super - hydrophobic setback surface prevents deflection of saline or biological fluids into the setback volume ( i . e . the surface meniscus is collinear with the ferrule output face or deflects outward from the ferrule output face ). a additional embodiment of the invention is presented in fig7 . a protective ferrule or ferrule 405 , similar to those discussed before , is produced with three bore diameters ; one bore 410 at approximately 450 μm accommodates the nylon or efte ( or hytrel or other common fiber optic buffer polymer ) diameter 415 of the fiber 400 , a second bore 420 at approximately 275 μm accommodates the fa coated fiber 425 , and a third bore 430 at approximately 350 μm accommodates a polished section of sapphire optical fiber 435 . the fiber 400 is glued 440 within the ferrule 405 within the buffer bore , the transition chamfer and the fa bore . the ferrule is half filleted 445 at each end for easy passage through a deflected ureteroscope . in the illustration , the fiber output surface 450 is slightly convex to quasi - collimate / focus the emerging laser light , although this is not a requirement for this embodiment . the themes of this invention remain the displacement of the water column between the fiber and the target for more efficient coupling of the laser energy to the target stone , and the avoidance of the rapid burn back phenomenon seen in small laser lithotripsy fibers that reduces surgical efficiency by approximately 40 % to 50 % within the first minute or two of surgery . while the devitrification issues are addressed in earlier embodiments through larger mass target contact components ( where silica ferrules are utilized ) or through the use of refractory and devitrification resistant , crystalline and polycrystalline ( ceramic ) materials , in the embodiment depicted in fig7 , the devitrification labile silica ferrule ( if silica ) is shielded from target contact by a slight protrusion 455 of the sapphire window as well . an additional embodiment , depicted in fig8 , simplifies the process of producing a closed - end 475 capsule 460 by forming the ferrule consisting entirely of sapphire ( as compared to fig7 ). the proximal 465 and distal 466 ends are , preferably , filleted ( or chamfered ) for smooth passage through the ureteroscope channel . the capsule end ( opposite the closure 475 ) is open 470 and of an appropriate diameter bore 480 to accommodate the fiber buffer diameter 500 . a second bore 485 is of an appropriate diameter to accommodate the fiber first polymer coating 495 . the output surface 490 of the fiber is depicted as curved for reduced divergence but may be flat . the ferrule &# 39 ; s second bore 485 , preferably , ends at a closed - end output region or lens 475 . the lens 475 preferably includes a flat interior surface ( adjacent to the borehole ) and either a flat or convex exterior surface . in one instance , the lens 475 includes a convex exterior surface that is congruous with the fillet 466 of the curved ferrule surface . in another instance , the fiber tip 490 has a convex lens that when combined with the exterior convex lens 475 on the sapphire ferrule , provides a collimated or approximately collimated beam immediately distal to the assembly for extremely efficient energy deliver to stone targets . that is , the fiber output surface and the ferrule lens cooperatively focus the beam on the surgical target . as in prior embodiments , the fiber can be retained within the capsule by means of adhesive within the bores . in another embodiment , depicted in fig9 , the ferrule can be affixed to the fiber with a crimp connector . this crimp connector can be employed in all embodiments that utilize non - metallic ferrules or caps ( described above ). for ease of explanation , fig9 depicts a sapphire ferrule 505 that is similar to the ferrule in fig8 but upon which a stepped and smaller outer diameter 510 is disposed . the smaller outside diameter 510 is preferably affixed within a borehole 515 of a metallic , crimp connector 530 , where the two mating surfaces are bonded by means known in the art ( e . g ., adhesive bonding ). the crimp connector , preferably , includes a second and smaller inside diameter 520 which accept the fiber buffer . in another instance , the crimp connector can have a single borehole ( single inside diameter ) where the sapphire ferrule has a smaller outside diameter that is approximately the same diameter as the fiber buffer outside diameter . the fiber is fixed within the ferrule 535 borehole by an adhesive while it is fixed within the crimp connector 520 by crimping 525 ( crimp indentations are shown absent the fiber ). the optical functionality remains unaltered with respect to the embodiment in fig8 , but the sapphire ferrule is , preferably , retained on the fiber by two separate mechanisms for security .

herein are disclosed protected optical fiber terminations for use in the treatment of renal and biliary calculi . the protected optical fiber termination including a ferrule affixed to the optical fiber that provides the termination of the optical fiber with protection from contact with saline and / or biological fluids . the optical fiber termination can include open or closed ferrules . the open ferrules providing a means for maintaining a bubble , e . g ., a moses bubble on the termination ; the closed ferrules providing a calumniated focus on the renal or biliary calculi .

as a preliminary matter , it will readily be understood by one having ordinary skill in the relevant art (“ ordinary artisan ”) that the present invention has broad utility and application . furthermore , any embodiment discussed and identified as being “ preferred ” is considered to be part of a best mode contemplated for carrying out the present invention . other embodiments also may be discussed for additional illustrative purposes in providing a full and enabling disclosure of the present invention . moreover , many embodiments , such as adaptations , variations , modifications , and equivalent arrangements , will be implicitly disclosed by the embodiments described herein and fall within the scope of the present invention . accordingly , while the present invention is described herein in detail in relation to one or more embodiments , it is to be understood that this disclosure is illustrative and exemplary of the present invention , and is made merely for the purposes of providing a full and enabling disclosure of the present invention . the detailed disclosure herein of one or more embodiments is not intended , nor is to be construed , to limit the scope of patent protection afforded the present invention , which scope is to be defined by the claims and the equivalents thereof . it is not intended that the scope of patent protection afforded the present invention be defined by reading into any claim a limitation found herein that does not explicitly appear in the claim itself . thus , for example , any sequence ( s ) and / or temporal order of steps of various processes or methods that are described herein are illustrative and not restrictive . accordingly , it should be understood that , although steps of various processes or methods may be shown and described as being in a sequence or temporal order , the steps of any such processes or methods are not limited to being carried out in any particular sequence or order , absent an indication otherwise . indeed , the steps in such processes or methods generally may be carried out in various different sequences and orders while still falling within the scope of the present invention . accordingly , it is intended that the scope of patent protection afforded the present invention is to be defined by the appended claims rather than the description set forth herein . additionally , it is important to note that each term used herein refers to that which the ordinary artisan would understand such term to mean based on the contextual use of such term herein . to the extent that the meaning of a term used herein — as understood by the ordinary artisan based on the contextual use of such term — differs in any way from any particular dictionary definition of such term , it is intended that the meaning of the term as understood by the ordinary artisan should prevail . furthermore , it is important to note that , as used herein , “ a ” and “ an ” each generally denotes “ at least one ,” but does not exclude a plurality unless the contextual use dictates otherwise . thus , reference to “ a picnic basket having an apple ” describes “ a picnic basket having at least one apple ” as well as “ a picnic basket having apples .” in contrast , reference to “ a picnic basket having a single apple ” describes “ a picnic basket having only one apple .” when used herein to join a list of items , “ or ” denotes “ at least one of the items ,” but does not exclude a plurality of items of the list . thus , reference to “ a picnic basket having cheese or crackers ” describes “ a picnic basket having cheese without crackers ”, “ a picnic basket having crackers without cheese ”, and “ a picnic basket having both cheese and crackers .” finally , when used herein to join a list of items , “ and ” denotes “ all of the items of the list .” thus , reference to “ a picnic basket having cheese and crackers ” describes “ a picnic basket having cheese , wherein the picnic basket further has crackers ,” as well as describes “ a picnic basket having crackers , wherein the picnic basket further has cheese .” referring now to the drawings , one or more preferred embodiments of the present invention are next described . the following description of one or more preferred embodiments is merely exemplary in nature and is in no way intended to limit the invention , its implementations , or uses . turning to fig1 - 2 , a cedar oil evaporator 100 in accordance with a first embodiment of the invention is illustrated . specifically , fig1 is a perspective view of the cedar oil evaporator 100 , and fig2 is an exploded view of the evaporator 100 . the evaporator preferably is intended for use in a dresser drawer . the evaporator 100 includes a circular container 102 having openings 104 formed in a top 106 thereof for the ventilation of air . the evaporator 100 also includes a large length of cotton twine 110 that has been saturated in cedar oil such that the cotton twine 110 disperses the cedar oil via the ventilated air . the cotton twine 110 is randomly disposed within the circular container 102 . in contrast to the other illustrated embodiments herein , no openings are formed in the bottom 108 of the container 102 , and the evaporator 100 does not include any reservoir of cedar oil . turning now to fig3 - 4 , a cedar oil evaporator 200 in accordance with a second embodiment of the invention is illustrated . specifically , fig3 is a perspective view of the cedar oil evaporator 200 , and fig4 is a perspective view similar to that of fig3 , wherein a door 206 of a housing 202 of the evaporator is open for showing the interior thereof . the cedar oil evaporator 200 preferably is intended for use in a closet for the protection of coats and clothes from moths . the cedar oil evaporator 200 includes openings 204 formed in the door 206 and in a back wall 208 of the rectangular housing 202 , which openings 204 are for the ventilation of air therethrough . the evaporator 200 further includes four separate lengths 210 of cotton twine . each length 210 of cotton twine extends from a bottle 212 of cedar oil ( or reservoir of cedar oil ) and includes a portion thereof that is immersed in the cedar oil within the bottle for wicking of the cedar oil out of the bottle . furthermore , each length 210 of cotton twine preferably has a great length with the portion of the cotton twine that is immersed in the bottle being a very small extent of the overall length 210 of the cotton twine . indeed , this immersed portion preferably represents 10 % or less of the overall length 210 of the cotton twine . the remainder of each of cotton twine length 210 is randomly contained within the interior space of the evaporator 200 ( i . e ., bunched as shown 214 in fig4 ) in such a manner that a large surface area of each of the cotton twine lengths 210 is exposed to air that passes through the ventilation openings 204 in the housing 202 . it will be appreciated from fig4 that the cotton twine lengths 210 define a plurality of interstices through which air can pass . it is believed that the cedar oil is wicked from each bottle 212 through each extent of the cotton twine lengths 210 and is dispersed via the ventilated air . a wire forms a hook 216 and extends from the top of the housing 202 of the evaporator 200 for the preferred hanging of the evaporator 200 in a closet . fig5 - 9 illustrate another cedar oil evaporator 300 in accordance with a third embodiment of the invention . specifically , fig5 is a perspective view of the cedar oil evaporator 300 ; fig6 is a perspective view similar to that of fig5 , wherein a door 306 of a housing 302 of the evaporator 300 is open for showing the interior thereof , fig7 is a perspective view similar to that of fig6 , wherein the cotton - twine wicks 310 have been omitted for clarity of illustration ; fig7 a is a perspective exploded view of certain of the components of the cedar oil evaporator 300 ; fig8 is a front elevational view of the cedar oil evaporator 300 further showing the interior thereof , and fig9 is a front elevational view similar to that of fig8 , wherein the cotton - twine wicks 310 again have been omitted for clarity of illustration . like the cedar oil evaporator 200 , the cedar oil evaporator 300 also is preferably intended for use in a closet . the cedar oil evaporator 300 includes openings 304 formed in the door 306 and in a back wall 308 of the housing 302 , which openings 304 are for the ventilation of air therethrough . the evaporator 300 further includes four separate lengths 310 of cotton twine . each length 310 of cotton twine extends from a single container or reservoir 312 of cedar oil ( or reservoir of cedar oil ). each length 310 serves as a carrier and includes a portion thereof that is immersed in the cedar oil within the reservoir 312 for wicking of the cedar oil out of the reservoir . furthermore , each length 310 of cotton twine preferably has a great length with the portion of the cotton twine that is immersed in the reservoir 312 being a very small extent of the overall length 310 of the cotton twine . indeed , this immersed portion preferably represents 10 % or less of the overall length 310 of the cotton twine . the remainder of each of cotton twine length 310 is randomly contained within the interior space of the evaporator 300 ( i . e ., bunched as shown 314 in fig6 and 8 ) in such a manner that a large surface area of each of the cotton twine lengths 310 is exposed to air that passes through the ventilation openings 304 in the housing 302 . it is believed that the cedar oil is wicked from the reservoir 312 through each extent of the cotton twine lengths 310 and is dispersed via the ventilated air . the evaporator 300 further includes a hook 316 that is attached to and extends from the top of the housing 302 of the evaporator 300 for the preferred hanging of the evaporator 300 in a closet . as will be appreciated from the foregoing , the cedar oil evaporator 300 is similar to evaporator 200 except principally in the inclusion of a single reservoir 312 of cedar oil rather than four separate reservoirs in the form of the bottles 212 . with further regard to the single reservoir 312 , the reservoir 312 includes a base 318 and a cover 320 with four openings 322 formed therein . each of the four cotton twine lengths 310 extends from a respective one of the opening 322 . in each of the illustrated evaporator 200 , 300 , each of the cotton twine lengths 210 , 310 ( also sometimes referred to as the carrier ) preferably is one - sixteenth of an ( 1 / 16 ) inch in diameter and includes an overall length of twelve and one - half ( 12½ ) feet . by comparison , the extent of each of the cotton twine lengths 210 , 310 that is immersed in cedar oil preferably is only a few inches . engineering drawings of a commercial embodiment of an evaporator in accordance with the invention , and representative of evaporator 300 , are illustrated in fig1 - 21 . these engineering drawing include preferred dimensions as shown therein . it will be appreciated by an ordinary artisan that when used with cotton twine lengths 310 which include an overall length of twelve and one - half ( 12½ ) feet as disclosed hereinabove , the overall length of each carrier will be an order of magnitude larger than the greatest dimension of the housing of the evaporator , i . e . seven and nine hundred and six thousandths ( 7 . 906 ) of an inch , which dimension is illustrated in fig1 . in use , the cotton twine lengths 210 , 310 may be immersed in cedar oil when the evaporators 200 , 300 are first installed in a closet whereby the cedar oil that initially evaporates therefrom is replaced with cedar oil wicked from a respective reservoir of the evaporators 200 , 300 . having now described in detail the illustrated evaporators , it is noted that a feature found in each of the three illustrated embodiments is the use of a carrier or wick that has a very large surface area that is exposed to air for the evaporation of cedar oil . indeed , with respect to the second and third illustrated embodiments , these devices with their reservoirs are intended to be used in closets for providing a sufficient amount of evaporation of cedar oil so as to provide an equivalent to new or revitalized cedar closets , i . e ., it is believed that the second and third illustrated embodiments provide the equivalent protection against moths as a new or revitalized cedar closet . with regard to the devices disclosed and described in the “ background of the invention ” section above , it is believed that the devices of travis and galler provide an insufficient surface area for evaporation of cedar oil so as to enable a sufficient amount of cedar oil to evaporate in order to provide emulation of a cedar closet over the course of several months . moreover , as cedar oil is believed to be much less volatile than perfumes and deodorizers , the prior art evaporators of u . s . pat . nos . 6 , 921 , 025 ; 4 , 621 , 768 ; and 4 , 352 , 457 are intended and designed for use with performs and deodorizers , and not with cedar oil , and therefore also are believed to provide an insufficient surface area for evaporation of cedar oil so as to enable a sufficient amount of cedar oil to evaporate in order to provide emulation of a cedar closet over the course of several months . based on the foregoing description , it will be readily understood by those persons skilled in the art that the present invention is susceptible of broad utility and application . many embodiments and adaptations of the present invention other than those specifically described herein , as well as many variations , modifications , and equivalent arrangements , will be apparent from or reasonably suggested by the present invention and the foregoing descriptions thereof , without departing from the substance or scope of the present invention . accordingly , while the present invention has been described herein in detail in relation to one or more preferred embodiments , it is to be understood that this disclosure is only illustrative and exemplary of the present invention and is made merely for the purpose of providing a full and enabling disclosure of the invention . the foregoing disclosure is not intended to be construed to limit the present invention or otherwise exclude any such other embodiments , adaptations , variations , modifications or equivalent arrangements , the present invention being limited only by the claims appended hereto and the equivalents thereof . for instance , while the carriers in the evaporators 200 , 300 have each been described as a length of cotton twine 210 , 310 , it is contemplated within the scope of the invention that these lengths of cotton twine may be separate and distinct from each other or , alternatively , may be part of a single length of cotton twine , or any combination thereof .

a cedar oil evaporator includes : a housing having openings therein for ventilation of air ; a reservoir within the housing containing cedar oil ; and a carrier for wicking away the cedar oil from the reservoir for dispersing cedar oil via air that passes through the housing . each carrier is a length of cotton twine having a diameter of one - sixteenth of an inch or smaller and a length of twelve and one - half feet or longer . the housing includes a door movable about a hinge between a first position , wherein the reservoir is accessible for removal from the housing ; and a second position , wherein the reservoir is inaccessible for removal . an extent of the carrier that is received within the reservoir for immersion in the cedar oil preferably represents 10 % or less of the overall length of the carrier , the remainder being randomly exposed within the interior space of the housing .

in one embodiment , the invention may be an agent infused cinnamon stick . this cinnamon stick will generally be at least semi - rigid , and have a substantially linear structure , a plurality of pores , and at least some water ( or food / beverage ) soluble cinnamon flavoring compounds . these pores will store , in a dry form , at least one chemical agent ( agent ), not normally found with cinnamon sticks . the overall structure and composition of the cinnamon stick will be such that when the cinnamon stick is immersed in a hot aqueous fluid ( e . g . a hot beverage ), the cinnamon stick will retain its substantially linear structure for at least a few minutes while the aqueous fluid ( hot beverage ) permeates the stick &# 39 ; s pores . the fluid will dissolve at least portions of the agent , and liberate it into the surrounding fluid along with usually at least some of the water soluble ( or food / beverage soluble ) cinnamon flavoring compounds that are normally associated with the cinnamon stick . in another embodiment , the invention may also be a method of producing chemical agent ( agent ) infused cinnamon sticks . this method will generally dissolving at least one agent in a carrier fluid , often an alcohol - water mixture , producing an agent solution , and applying this agent solution to usually many cinnamon sticks at the same time . the agent solution will be allowed to permeate the cinnamon sticks , excess agent solution removed as needed , and then the carrier fluid ( e . g . the alcohol - water mixture ) itself removed ( usually by a drying process ), thus producing dry cinnamon sticks infused with at least one agent . although , to simplify reading , most discussion will relate to flavor - infused cinnamon sticks for hot beverages , and various methods of making the same , the use of “ flavor ” or “ beverage ” is not intended to be limiting . in other embodiments , other functional ingredients ( agents ), such as vitamins , coloring agents , sweetening agents , and the like can also be infused into cinnamon sticks using the methods of the present invention . the sticks in turn can be used to flavor a wide range of hot and cold liquid beverages , semi - solid foods ( e . g . yogurt , sauces , etc .) or solid foods as well . at an abstract level , the invention may be described using the common technical problem , solution to problem format . currently there is a commercial need ( market demand ) for natural healthy ingredients for various foods and beverages . although cinnamon sticks are known to be a useful vehicle for imparting a cinnamon flavor , cinnamon sticks in their current format otherwise are limited in the sense that they are not used for anything beyond this limited purpose . as previously discussed , the invention is based , in part , on the concept of using cinnamon sticks as a delivery mechanism for other agents . here for example , the cinnamon stick as a delivery platform concept can be used to expand this platform to new market sectors by adding other flavors and healthy ingredients like vitamins . this would allow agent infused or modified cinnamon sticks to appeal to a wider cross - section of the population . in addition to consumers interested in novel flavors , the platform could also address various health market segments , such as segments focused on healthy aging , diabetes and cardiovascular disease , proper nutrition , and the like . to do this , in addition to teaching the concept of agent infused or modified cinnamon sticks as one embodiment of the invention , in another embodiment the invention teaches methods of producing such modified cinnamon sticks that , in a preferred embodiment , allow various agents that impart flavor , color , vitamins or other materials to be incorporated into the cinnamon stick structure in a manner that preserves the inherent length and stiffness of the cinnamon stick , preserves the integrity of the agent , and also allows both the agent and at least traces of cinnamon flavor to elude from the stick over a short ( few second to few minutes ) period of time . although many methods to produce the modified cinnamon sticks are possible , based on experimentation , it has been found that vacuum infusion methods have certain advantages . according to the vacuum infusion method ( illustrated in fig3 ), the cinnamon stick quills can be placed in an airtight infusion container in contact with the infusion fluid ( containing the dissolved agents ), but not completely immersed in the infusion fluid . air is then removed from the container , and the fluid moves into the various porous voids in the cinnamon stick . this approach has the advantage that the infusion takes place relatively evenly between the various cinnamon sticks , the infusion process proceeds rapidly , and due to the vacuum ( or low air pressure , such as less than 20 % of normal atmospheric pressure ), the infusion fluid travels deeply into the porous cinnamon stick matrix . this helps lock in more of the agent &# 39 ; s flavors and aromas , and also helps prevent surface evaporation from hindering flow of the agent into the deeper pores . this method is also good for forcing infusion of some hydrophobic substances ( agents ), such as oil based extracts , that otherwise would tend not to be absorbed into the hydrophilic cinnamon stick matrix . according to the vacuum method , and as shown in fig3 , in step ( 300 ), the cinnamon sticks may be placed in a vacuum chamber , and the air at least partially removed . in step ( 302 ), the cinnamon sticks may be exposed to the liquid dissolved agents under low pressure , and the liquid will then flow into the various pores of the cinnamon stick , depositing the agents deep into the cinnamon stick structure as desired . in step ( 304 ), the remaining liquid dissolved agents are removed from the chamber . in step ( 306 ), the sticks may be dried further under low pressure , and optionally then further treated with heat and / or treated with various stabilizing agents as will be described shortly . although vacuum infusion methods are ideal , other infusion methods , such as spray application ( illustrated in fig2 ), or even soaking the cinnamon sticks in the infusion fluid in the absence of a vacuum may also be used . additionally , methods such as enrobing , spray and tumble methods ( illustrated in fig4 ) and the like may also be used . indeed essentially any method to saturate the sticks with a fluid containing the agents may be used . before the cinnamon stick can be infused with the desired chemical agent or agents , the agents should first be dissolved into a carrier fluid . here use of pure water carrier fluids , although allowable , are not always preferred because over time , water can tend to weaken the structure of the cinnamon stick , and even cause tightly rolled cinnamon sticks , for example , to open up and lose their shape to some extent . as a result , in a preferred embodiment , often the carrier fluid will instead often be based on alcohol - water mixtures . thus for example , powdered agents ( e . g . powdered flavors , vitamins , sweetening agents ( e . g . sucralose ) and the like will often be dissolved , preferably at or near the highest possible concentration , in an alcohol - water mix prior to the start of the infusion process . generally the goal is to use concentrated agents so as to deliver a maximum amount of agent per unit amount of carrier fluid , thus minimizing the damage that the carrier fluid will cause to the cinnamon sticks during the infusion process . in some embodiments , these methods will result in up modified cinnamon sticks that carry as much as 5 to 10 % agents by weight ( e . g . relative to the weight of the cinnamon stick ). in some embodiments of the invention , it is also useful , after the initial infusion process , to then perform subsequent operations to help seal the flavors and aromas of the agent into the stick , thus promoting a good shelf life and optionally also facilitating subsequent release of the agent when the stick is subsequently immersed in a food or beverage , such as a hot water based beverage . here various sealing methods may be used . for example , in some embodiments , the outside of an infused and dried cinnamon stick quill may be sprayed with an aerosol - like glycerin mix , and then dried again . this results in a thin layer of glycerin surrounding the cinnamon stick , and helps promote a longer ( e . g . year or more ) shelf life for the final product . for an 8 gram cinnamon stick , often 0 . 2 to 0 . 4 grams of glycerin may be applied using this method . other stabilizing agents may also be used . types of flavorings : in addition to various herbal flavorings , many other types of flavorings may also be imbedded or infused into the cinnamon stick according to the invention . some of these other flavorings include vanilla , mint or peppermint , apple , orange , pumpkin , butter , or various natural or artificial sweeteners . many of the agents may be chosen so as to be suitable for herbal tea - like beverage . herbal tea is often a relatively simple combination of boiling water and dried teas , fruits , flowers , herbs and / or the like . one of the main ingredients of many herbal tea varieties is cinnamon . for example , cinnamon ( or cinnamon flavoring ) can be used in varieties of herbal beverages such as : apple cider , indian chai , citrus blend beverages and the like . however , in conventional teas , the cinnamon may be a separate ingredient in a teabag , pouch or filter . according to the invention , however , the soluble herbal tea substances may instead be concentrated and applied to the cinnamon stick . the net result can be a modified cinnamon stick that either produces a flavored herbal tea - like beverage when immersed in hot water by itself , or a modified cinnamon stick that can be used to supplement the flavor of a first herbal tea with a second set of flavors intended to complement the flavor of the first herbal tea . depending on the market in question , the agents may be deposited on the outside of the cinnamon stick , and / or in the central core of the cinnamon stick . as previously discussed , impregnation and drying can often be facilitated by use of vacuum methods . the main goal of the process is to produce a robust product that can be easily packaged into wrapped single cinnamon stick units ( e . g . for hotel / restaurant use ) or which can be easily packaged into multiple units ( e . g . retailers , home use ). the final product can thus be used and sold in a wide range of food and beverage applications such as for use in sweet or savory foods or beverages , alcoholic and non - alcoholic beverages , dairy foods and beverages , confectionary foods , and the like . fig1 shows a process ( 10 ) for making flavor - infused cinnamon sticks . the process ( 10 ) begins at ( 12 ) and continues to ( 14 ). at step ( 14 ), one or more cinnamon sticks are provided . the cinnamon sticks can be of the same length , e . g ., about 3 inches long . the process continues to ( 16 ). at step ( 16 ), a flavoring compound ( e . g ., an herbal extract concentrate ) is provided . the herbal extract can include one or more herbal extract flavoring ( or aroma ) concentrates . in addition to herbs , the extracts can include extracts of other plants , fruits , vegetables or the like . the flavoring compound can be natural flavoring or aroma compounds or artificial flavoring or aroma compounds , or a combination of the above . the process continues to ( 18 ). at step ( 18 ), the flavoring compound is applied to a cinnamon stick . the flavoring compound can be applied to the inside or outside ( or both ) of the cinnamon stick . a predetermined amount of flavoring can be used per stick . for example , an average cinnamon stick may weigh about 5 grams and the flavoring compound can comprise about 3 - 6 % of the finished product by weight . the process continues to ( 20 ). at step ( 20 ), the flavoring compound is allowed to optionally soak into the cinnamon stick for a predetermined period of time . the process continues to ( 22 ). at step ( 22 ), the cinnamon sticks with flavoring compounds applied are placed into a container with sticks of the same flavor , the container can be sealed and the infusion process can be allowed to continue during a second predetermined period of time ( e . g ., about 24 hours or more ). the process continues to ( 24 ). at step ( 24 ), the flavor - infused cinnamon sticks are ready for use or packing for sale . fig2 shows this process in more detail . here step ( 14 ) is shown in more detail as ( 200 ). as previously discussed for step ( 14 ), the unmodified cinnamon sticks are provided , preferably all approximately 3 inches long and all approximately the same length . step ( 16 ) is shown in more detail in ( 202 ). here the flavoring compound , such as the herbal extract concentrate , is provided as previously described . steps ( 18 ) and ( 20 ) are shown in more detail in ( 204 ). here the flavoring compound ( here an herbal extract concentrate ) is applied to the cinnamon sticks as previously described . a number of different methods may be used to apply this flavoring compound , as described elsewhere . step ( 24 ) is shown in more detail in ( 206 ). here the flavor infused cinnamon sticks are shown packed and ready for resale and use . as previously discussed , although vacuum infusion methods are particularly useful , many other methods may also be used to produce the modified cinnamon sticks . some of these methods include dipping , enrobing , vacuum tumbling or spraying , electrostatic spraying etc . examples of the equipment used for the enrobing and spray and tumbling methods are shown in fig4 . fig4 shows an example of enrobing equipment ( 400 ), which may be used in a manner similar to those used by chocolate filled bar manufacturers ), and spray and tumble equipment ( 402 ). an embodiment includes an infused cinnamon stick with flavor . the infused flavors can be single flavors or combinations of flavors . for example , to infuse cinnamon sticks with flavors from each recipe , varied amounts of natural herbal extracts of the respective flavors , or very similar ones can be used to infuse the cinnamon sticks for each respective flavoring recipe . another embodiment would be a cinnamon stick infused with a functional ingredient such as a vitamin . infusion of herbal flavors can be accomplished using oil or alcohol based herb ( or other flavoring ) extracts . the infusion flavorings can be natural or artificial flavorings . natural herbal extracts are typically a preparation in which the active flavor or aromatic compounds of an herb are extracted into alcohol ( or oil ). this process concentrates and preserves the qualities of the herb flavor . extracts are sold commercially , and used specifically to create soft drinks and beverages , for example . an embodiment can include a process to manufacture a flavor - infused cinnamon stick . for example , the process can include providing a cinnamon stick and applying a predetermined amount of concentrated extract flavoring ( one or two drops ) on the inside ( the lumen ) and / or outside of a cinnamon stick ( a cinnamon stick typically 3 inches in length but other sizes can be used depending on the market application ), and is left for a predetermined time period ( typically a few seconds ) to absorb into the woody part of the cinnamon stick . the stick is then left for another predetermined time period ( e . g ., about 24 hours or less if vacuum applied ) to facilitate the infusion process in a closed container with other sticks of the same flavor recipe . the flavor or flavors can be further fixed into place using edible adhesives based on carbohydrates and gums . take 2 g of natural flavor ( in 0 . 4 g of alcohol ) and add to 8 g of cinnamon sticks . leave for 12 hours . the process conditions would change and are dependent on the type of adduct or flavor to be infused into the cinnamon stick and the degree to which the desired effect is required . a similar process could be used to incorporate functional health ingredient such as vitamins , antioxidants , natural edible colors , sweeteners , or bioactive ingredients etc . other treatments can also include fixing the agent or adduct in the cinnamon stick either chemically or with heat . this will protect the product and aid release only when the cinnamon stick is in use . another embodiment can include a process to add edible dyes , inclusions and other ingredients to the cinnamon stick . for example : vitamins or ingredients that relieve upset stomach and agents that can boost energy . in use , the adduct - infused cinnamon stick can be placed in boiling or hot water so that the cinnamon releases the flavor of the cinnamon along with the flavors of the infused flavors ( e . g . herbal extracts ) to create a flavor and aroma of a particular recipe . the flavor and aroma will depend on the extracts used in the recipe . the cinnamon stick is typically not eaten , only placed into a drink or food dish to impart flavors and / or aromas . however , because of its versatile flavoring , a flavor - infused cinnamon stick can be used for hot or cold drinks , and also for sweet or savory foods , such as soups , rice , beef casseroles , or the like . for the functional health ingredients a similar process is recommended where the cinnamon stick infused with ingredient is placed in hot or boiling water . it can also be envisioned that this invention can be used in cold drinks for example alcoholic and non - alcoholic cocktails . during the development of prototypes , 120 infusion drinkers were given samples to taste in a simple test . three samples were rated for naturalness , flavor and aroma . the three samples were as follows : ( i ) a cinnamon flavor infused stick ( ii ) a control commercial loose tea infusion from company a and ( iii ) a control commercial loose tea infusion from company b . the cinnamon infused stick was matched as closely as possible in previous trials to match the commercial samples . the modified cinnamon stick was rated first for naturalness , flavor and aroma . the experiment was conducted under controlled conditions for example tea bags and cinnamon sticks were removed from the samples prior to presenting to the panels to avoid any visual bias . it is , therefore , apparent that there is provided , in accordance with the various embodiments disclosed herein , flavor - infused cinnamon sticks and a method of making the same . while the invention has been described in conjunction with a number of embodiments , it is evident that many alternatives , modifications and variations would be or are apparent to those of ordinary skill in the applicable arts . accordingly , the applicant intends to embrace all such alternatives , modifications , equivalents and variations that are within the spirit and scope of the invention .

flavor infused cinnamon sticks and a method for making the same . natural cinnamon sticks are infused with various flavors , flavor pre - cursors , or other agents such as vitamins , sweeteners , or food colorants . the cinnamon sticks provide a natural matrix for carrying the various flavors or other agents , as well as providing a convenient handheld stirrer when used to add flavor to a beverage . although ground cinnamon produces an intense amount of cinnamon flavor that would dominate over other infused flavors , the cinnamon flavor in intact cinnamon sticks leaches out only slowly , and thus the cinnamon flavor can harmoniously co - exist with the various infused flavors . various methods of producing the infused cinnamon sticks are also taught .