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A technique for the instillation of solutions into the rat uterine lumen is described. The method has been tested by following the receptor depletion/replenishment cycle after oestradiol injection and by checking on the stoichiometry of hormone/receptor translocation from the cytoplasmic compartment into the nucleus. Both crude sediments from frozen uteri and nuclei purified by a novel procedure were analyzed and gave identical results. The limitations and the advantages of the technique are discussed.

22 patients entered a double-blind trial to test the efficacy of bromocriptine therapy in patients with Parkinson's disease already established on conventional levodopa therapy. 3 patients on placebo withdrew when no improvement occurred and control became complicated. 4 patients on active drug withdrew because of various symptoms, but in only 1 case were these (nausea and vomiting) thought to be a real drug effect. Of the 15 patients who completed the trial, 9 were on active drug and 6 were on placebo. Although more than half the patients in each group were subjectively improved, measurement scales of functional disability and physical examination revealed no significant change in either group. Side effects encountered included nausea, dyskinesiae and hallucinations. It is concluded that bromocriptine does not offer any additional benefit in most patients with Parkinson's disease who are well stabilised on levodopa therapy, but may have a place in those patients who encounter side effects due to fluctuations in serum and tissue levels of levodopa.

OBJECTIVE: To determine whether injuries to sub-optimally restrained child occupants in real-world crashes were likely to be preventable by alternative restraint usage practices and to assess the usefulness of crash reconstruction for exploring injury mechanisms in child occupants.METHODS: Real-world crashes in which child occupants sustained significant injuries were reconstructed on a laboratory crash sled using the Hybrid III family of child dummies. Alternative restraint scenarios and cases in which children were not seriously injured were also simulated to compare dummy kinematics and dynamic responses in optimal restraint configurations.RESULTS: Restraint misuse was associated with greater motion of the dummy torso and head during crashes, often allowing contact between the child and the vehicle interior, resulting in injury. Poor pre-crash posture for a child inappropriately restrained in an adult belt appeared to worsen the geometry of the sash (shoulder) belt, resulting in a cervical injury due to direct interaction with the belt. Dynamic dummy data did not appear to discriminate between injury and non-injury cases.CONCLUSIONS: Dummy kinematics suggest that injuries in which inappropriate use and misuse were a factor were less likely if the most appropriate restraint was used correctly. Adequately controlling the head and upper body of the child occupant was seen to prevent undesirable interactions with the vehicle interior and restraint system, which were associated with injury in the real world. Neck forces and moments and injury criteria calculated from these did not predict injury reliably.

Trimethoprim-sulfamethoxazole (TMP-SMX) medication in the feed or water is commonly administered to immunocompro mised mice to prevent the occurrence of Pneumocystis murina (formerly P. carinii) pneumonia. Therapeutic doses of SMX can cause decreased total and free thyroxine (T4) levels in dogs and thyroid hypertrophy and hyperplasia in mice, rats, and dogs. Our primary objective was to determine whether SMX at doses present in commercially available rodent TMP-SMX feed would pro duce hypothyroidism in mice. Plasma T4 levels were determined prior to and after placement of Brand A TMP-SMX feed (daily SMX dose, 240 mg/kg), Brand B TMP-SMX feed (daily SMX dose, 2400 mg/kg), and their respective controls (doses calculated for a 25-g mouse according to vendor's information). T4 levels in the mice fed Brand B TMP-SMX feed were significantly decreased by 2 wk after feed placement. Levels of thyroid stimulating hormone in male and female mice given Brand B TMP-SMX feed were significantly elevated compared with those of control groups at 6 wk after feed placement, when only these mice showed evidence of thyroid hypertrophy and hyperplasia. No significant change in T4 levels occurred over the course of 11 wk in mice given the Brand A TMP-SMX chow or either control feed. In light of the significant clinical hypothyroidism that occurred in our mice while receiving Brand B TMP-SMX diet, we recommend SMX levels more similar to that of Brand A to avoid such unwanted effects which could confound research data.

The relationship was investigated between different ultrasonographically defined subtypes of the human corpus luteum and progesterone production. Twenty-one women in the mid-luteal phase who underwent laparotomy for benign uterine conditions volunteered for this study. The corpus luteum was identified by preoperative ultrasound and classified into four types according to earlier established criteria, where types a and c were centrally hypoechoic, types b and d were centrally echogenic, types a and b had thin surrounding 'walls' (<3 mm) and types c and d had thick walls (<3 mm). After luteectomy, the theca externa capsule was removed and tissue from directly beneath the surface ('peripheral region') and the layer immediately beneath ('inner region') minced into 4-6 mg pieces. Following preincubation, pieces were incubated for 3 h at 37 degrees C in HEPES-minimal essential medium buffer with or without human chorionic gonadotrophin (HCG; 10 IU/ml), and subsequently progesterone accumulation in the medium was determined by radioimmunoassay. The highest progesterone production was consistently seen in the peripheral region. Type a had a significantly (P > 0.01) lower progesterone production (3.2 +/- 1.5 nmol/g tissue wet weight, mean +/- SEM, n = 4) than that of types b, c and d (17.7 +/- 3.5 nmol/g tissue wet weight, n = 9). All types responded to HCG with an almost two-fold increase in progesterone production. However, the maximal progesterone produced following stimulation by HCG in the type a corpus luteum was <50% of the basal (unstimulated) progesterone synthesis of any other type of corpus luteum. Using in-situ hybridization, with a primate RNA probe complementary to the region coding the extracellular part of the luteinizing hormone (LH) receptor, a highly localized expression of LH receptor mRNA to the peripheral region was found. Negligible or low levels of expression were found in the theca externa capsule and the inner region. No obvious correlations between the different subtypes of corpora lutea and LH receptor mRNA expression were seen. Thus, the ultrasonographic detection of a thin-walled and centrally hypoechoic corpus luteum correlates well with reduced progesterone secretion. The underlying cellular mechanism does not appear to involve a diminished sensitivity to the gonadotrophic stimulation by LH or HCG.

BACKGROUND: Seborrheic dermatitis is an inflammatory dermatosis with a prevalence of 1-3% in the general population. It is recognized more commonly in those infected by human immunodeficiency virus (HIV). No studies have looked at Seborrheic dermatitis in the context of immunosuppression secondary to organ transplantation. Therefore, we investigated the prevalence of Seborrheic dermatitis in a renal transplant population and characteristics of those affected.METHODS: A prospective study of 308 renal transplant recipients (RTRs) was carried out. All participants were examined for Seborrheic dermatitis. Descriptive statistics were employed and associations with Seborrheic dermatitis were examined using Fisher's exact test to calculate P-exact values, and Student's t-test was used to compare mean ages and time since transplantation. Statistical analysis was carried out using SPSS version 14.0 for Windows.RESULTS: Seborrheic dermatitis was identified in 29/308 (9.5%) patients and was more common in males (P-exact = 0.004) and in those who had been transplanted for longer (P = 0.02). The disease was mild-moderate severity in the majority but an unusual flexural appearance was recorded in 7/29 patients. Seborrheic dermatitis was less likely in those taking prednisolone (P-exact = 0.006) or tacrolimus (P-exact = 0.008). Seborrheic dermatitis was significantly associated with cutaneous malignancy, in particular squamous cell carcinoma (P-exact < 0.0001).DISCUSSION: Seborrheic dermatitis is more common than other inflammatory dermatoses in immunosuppressed RTRs, but is not as frequent as in those immunosuppressed secondary to HIV. Degree and duration of exposure to immunosuppression and increased colonization with Malassezia yeast genus are likely be important in the aetiology of Seborrheic dermatitis in RTRs. Further studies are required to clarify this.

UNLABELLED: A retrospective series of 99 femoral shaft fractures treated by small diameter (10 and 11 mm) and large diameter (> 11 mm) closed section femoral nails from November 1989 to September 1993 was analyzed. No significant differences in the parameters of bony union and time to full weight bearing were seen between the two groups nor were there significant differences between the rate of secondary procedures. There were no broken nails in either group and there was no difference in the overall respiratory complication rate.CONCLUSION: No statistically significant differences existed between the small and large diameter groups except for the mean age and mean follow-up period. Small diameter nails can be used safely without the risk of nail breakage.

In this work MRI-based spirometry is presented as a method for noninvasively assessing pulmonary mechanical function on a regional basis. A SPAMM tagging sequence was modified to allow continuous dynamic imaging of the lungs during respiration. A motion-tracking algorithm was developed to track material regions from time-resolved grid-tagged images. Experiments were performed to image the lungs during quiet breathing and volumetric strain was calculated from the measured displacement maps. Regional volume calculations, derived from volumetric strain, were integrated over the entire lung and compared to segmented volume calculations with good agreement. Results from this work demonstrate that MRI spirometry has the potential to become a clinically useful tool for measuring regional ventilation and assessing pulmonary diseases that regionally affect the mechanical function of the lung.

A model of the trichodiene synthase (TRI5) of the wheat fungal pathogen and type-B trichothecene producer Fusarium culmorum was developed based on homology modelling with the crystallized protein of F. sporotrichioides. Eight phenolic molecules, namely ferulic acid 1, apocynin 2, propyl gallate 3, eugenol 4, Me-dehydrozingerone 5, eugenol dimer 6, magnolol 7, and ellagic acid 8, were selected for their ability to inhibit trichothecene production and/or fungal vegetative growth in F. culmorum. The chemical structures of phenols were constructed and partially optimised based on Molecular Mechanics (MM) studies and energy minimisation by Density Functional Theory (DFT). Docking analysis of the phenolic molecules was run on the 3D model of F. culmorum TRI5. Experimental biological activity, molecular descriptors and interacting-structures obtained from computational analysis were compared. Besides the catalytic domain, three privileged sites in the interaction with the inhibitory molecules were identified on the protein surface. The TRI5-ligand interactions highlighted in this study represent a powerful tool to the identification of new Fusarium-targeted molecules with potential as trichothecene inhibitors.

The fifth ?land Island meeting on von Willebrand disease (VWD) was held on the ?land Islands, Finland, from 22 to 24 September 2016-90 years after the first case of VWD was diagnosed in a patient from the ?land Islands in 1926. This meeting brought together experts in the field of VWD to share knowledge and expertise on current trends and challenges in VWD. Topics included the storage and release of von Willebrand factor (VWF), epidemiology and diagnostics in VWD, treatment of VWD, angiogenesis and VWF inhibitors.

Mica is proposed as a convenient substrate for SEM specimens. Mechanically, mica is rigid and can be cut, punched and cleaved into suitable sizes. Compared to glass substrates, mice seems to be more charge 'resistant' when irradiated with the same beam density in the microscope.

The regio- and stereoselectivity of warfarin metabolism have been used to assess structure-function relationships of human P4502C subfamily members. Metabolism was investigated using a yeast cDNA expression system in which full length cDNAs for P4502C8, -2C9 (alleles Arg144 Tyr358 Ile359 Gly417 and Arg144 Tyr358 Leu359 Gly417), -2C18 (alleles Thr385 and Met385), and -2C19 were expressed. Additionally, two mutations reported in other P4502C9/2C10 alleles were individually introduced into P4502C9 by site-directed mutagenesis, to yield Cys144 Tyr358 Ile359 Gly417, Arg144 Tyr358 Ile359 Asp417, and Arg144 Cys358 Ile359 Gly417, which were expressed in yeast; their ability to metabolize warfarin was then studied. Warfarin metabolism by purified preparations of P4502C9 allele Arg144 Tyr358 Ile359 Gly417 and its Leu359 mutant was also investigated in reconstituted systems. Both alleles of P4502C18 were regioselective for 4'-hydroxywarfarin, without any significant stereoselectivity. Both also metabolized warfarin at the 6-position, but to a lesser extent, and metabolism at this site was stereoselective for (R)-warfarin. P4502C8 metabolized warfarin at the 7-position and was stereospecific for (R)-warfarin. It also metabolized warfarin to a lesser extent at the 4'-position, and metabolism at this site was stereoselective for (R)-warfarin. P4502C19 was regioselective for 6- and 8-hydroxywarfarin and was stereoselective for (R)-warfarin. The highly conservative mutation of Ile359 to Leu359 in P4502C9 profoundly altered the regio- and stereoselectivity of warfarin metabolism, from regioselective for 7-hydroxywarfarin, with stereospecificity for (S)-warfarin, to regioselective for 4'-hydroxywarfarin, with stereoselectivity for (R)-warfarin, which was confirmed in a reconstituted system using purified recombinant enzymes. In contrast, individual mutations of P4502C9 of Arg144 to Cys, Tyr358 to Cys, and Gly417 to Asp did not markedly affect the regio- or stereoselectivity of warfarin metabolism, although the overall rates of warfarin metabolism were apparently increased by these changes. We conclude that residue 359 is at the substrate binding site of P4502C9, whereas residues 144, 358, and 417, and residue 385 of P4502C18, are not.

Individuals of different quality may have different investment strategies, shaping responses to experimental manipulations, thereby rendering the detection of such patterns difficult. However, previous clutch-size manipulation studies have infrequently incorporated individual differences in quality. To examine costs of incubation and reproductive investment in relation to changes in clutch size, we enlarged and reduced natural clutch sizes of four and five eggs by one egg early in the incubation period in female common eiders (Somateria mollissima), a sea duck with an anorectic incubation period. Females that had produced four eggs (lower quality) responded to clutch reductions by deserting the nest more frequently but did not increase incubation effort in response to clutch enlargement, at the cost of reduced hatch success of eggs. Among birds with an original clutch size of five (higher quality), reducing and enlarging clutch size reduced and increased relative body mass loss respectively without affecting hatch success. In common eiders many females abandon their own ducklings to the care of other females. Enlarging five-egg clutches led to increased brood care rate despite the higher effort spent incubating these clutches, indicating that the higher fitness value of a large brood is increasing adult brood investment. This study shows that the ability to respond to clutch-size manipulations depends on original clutch size, reflecting differences in female quality. Females of low quality were reluctant to increase investment at the cost of lower hatch success, whereas females of higher quality apparently have a larger capacity both to increase incubation effort and brood care investment.

In adults, hemangioma is a rare presentation of a parotid mass. In this report, we present a case of a 40-year-old lady complaining of a swelling in the right inferior parotid region. Imaging studies revealed a cystic lesion of the right parotid gland. Fine needle aspiration cytology was not significantly helpful in revealing the diagnosis. In the operating room, a vascular cystic lesion was found in the deep lobe of the right parotid gland. Pathological analysis confirmed the diagnosis of cavernous hemangioma. In this case report, we discuss a rare pathology in the adult population, and review different aspects in its diagnosis and treatment.

The furcation regions of 30 human mandibular molars were perforated and sealed using four different materials: IRM, light- and chemically-cured glass ionomer cement (GIC), and amalgam. The materials were compared for marginal gaps in coronal, mid, and apical regions after routine SEM processing. While light-cured GIC showed the smallest gaps in the three regions, in mid and coronal regions chemically-cured GIC, and in apical regions amalgam, showed the largest gaps. IRM cases showed the highest rate of fillings with a good "fit", whereas the majority of amalgam cases and none of the chemically-cured GIC cases were overfilled.

In order to elucidate the still poorly understood pathogenetic pathways of acute tinea, the inflammatory cellular infiltrates in this infection were analyzed. Lesional punch biopsies were cryostat-sectioned and stained with monoclonal antibodies for immunophenotypization of T cells, B cells, macrophages and activation markers. For each antibody the positively stained inflammatory cells in the dermis and in the epidermis were quantified separately. Most of the dermal mononuclear cells in acute tinea were identified as T helper lymphocytes of the memory type. Furthermore, considerable amounts of Langerhans' cells and macrophages were found, but virtually no B cells. A high proportion of cells expressed markers of activation. Within the epidermis, accumulations of Langerhans' cells and LeuM5+ dendritic macrophages were detected near fungal element. In view of the otherwise rather similar cellular infiltrates in acute tinea and different non-infectious dermatoses, acute tinea may be particularly suitable to study the functional relationship of Langerhans cells and LeuM5+ macrophages.

This grounded theory study explored parents' experiences of responding to their children's need for understanding parental mental health concerns. Fifteen parents with severe and enduring mental health difficulties participated in the study. The findings suggest four main social processes that influence parents' talk with their children about parental mental health issues, namely "Protecting and being protected," "Responding to children's search for understanding," "Prioritizing family life," and "Relating to others." Implications of the findings for clinical practice and future research are considered. In particular, the need for more family-orientated services where parents experience parental mental health problems is highlighted.

We have studied the effect of macromolecular crowding reagents, such as polysaccharides and bovine serum albumin, on the refolding of tetradecameric GroEL from urea-denatured protein monomers. The results show that productive refolding and assembly strongly depends on the presence of nucleotides (ATP or ADP) and background macromolecules. Nucleotides are required to generate an assembly-competent monomeric conformation, suggesting that proper folding of the equatorial domain of the protein subunits into a native-like structure is essential for productive assembly. Crowding modulates GroEL oligomerization in two different ways. First, it increases the tendency of refolded, monomeric GroEL to undergo self-association at equilibrium. Second, crowding can modify the relative rates of the two competing self-association reactions, namely, productive assembly into a native tetradecameric structure and unproductive aggregation. This kinetic effect is most likely exerted by modifications of the diffusion coefficient of the refolded monomers, which in turn determine the conformational properties of the interacting subunits. If they are allowed to become assembly-competent before self-association, productive oligomerization occurs; otherwise, unproductive aggregation takes place. Our data demonstrate that the spontaneous refolding and assembly of homo-oligomeric proteins, such as GroEL, can occur efficiently (70%) under crowding conditions similar to those expected in vivo.

The cattle tick, Rhipicephalus (Boophilus) microplus, is a major threat to the improvement of cattle production in tropical and subtropical countries worldwide. Bos indicus cattle are naturally more resistant to infestation with the cattle tick than are Bos taurus breeds, although considerable variation in resistance occurs within and between breeds. It is not known which genes contribute to the resistant phenotype, nor have immune parameters involved in resistance to R. microplus been fully described for the bovine host. This study was undertaken to determine whether selected cellular and antibody parameters of the peripheral circulation differed between tick-resistant Bos indicus and tick-susceptible Bos taurus cattle following a period of tick infestations. This study demonstrated significant differences between the two breeds with respect to the percentage of cellular subsets comprising the peripheral blood mononuclear cell population, cytokine expression by peripheral blood leukocytes, and levels of tick-specific immunoglobulin G1 (IgG1) antibodies measured in the peripheral circulation. In addition to these parameters, the Affymetrix bovine genome microarray was used to analyze gene expression by peripheral blood leukocytes of these animals. The results demonstrate that the Bos indicus cattle developed a stabilized T-cell-mediated response to tick infestation evidenced by their cellular profile and leukocyte cytokine spectrum. The Bos taurus cattle demonstrated cellular and gene expression profiles consistent with a sustained innate, inflammatory response to infestation, although high tick-specific IgG1 titers suggest that these animals have also developed a T-cell response to infestation.

The sorting of integral membrane proteins such as carboxypeptidase S (Cps1p) into the luminal vesicles of multivesicular bodies (MVBs) in Saccharomyces cerevisiae requires ubiquitination of their cytosolic domains by the ubiquitin ligases Rsp5p and/or Tul1p. An exception is Sna3p, which does not require ubiquitination for entry into MVBs. The mechanism underlying this ubiquitination-independent MVB sorting pathway has not yet been characterized. Here, we show that Sna3p sorting into the MVB pathway depends on a direct interaction between a PPAY motif within its C-terminal cytosolic tail and the WW domains of Rsp5p. Disruption of this interaction inhibits vacuolar targeting of Sna3p and causes its accumulation in a compartment that overlaps only partially with MVBs. Surprisingly, Sna3p does require a functional ubiquitin-ligase HECT domain within Rsp5p; however, the dependence of Sna3p on HECT domain activity is distinct from that of Cps1p. Last, we show that Sna3p requires neither Tul1p nor the transmembrane adaptor protein Bsd2p for its MVB sorting. Our data demonstrate that Sna3p follows a novel ubiquitination-independent, but Rsp5p-mediated, sorting pathway to the vacuole.

A workshop was organised to investigate whether risk assessment strategies and methodologies used in classical/conventional toxicology may be used for risk assessment of allergenic foods, to discuss the advantages and limitations of different approaches and to determine the research needed to move the area forward. Three possible approaches to safety assessment and risk assessment for allergenic foods were presented and discussed: safety assessment using NOAEL/LOAEL and uncertainty factors, safety assessment using Benchmark Dose and Margin of Exposure (MoE), and risk assessment using probabilistic models. The workshop concluded that all the three approaches to safety and risk assessment of allergenic foods should continue to be considered. A particular strength of the MoE and probabilistic approaches is that they do not rely on low-dose extrapolations with its inherent issues. Probabilistic modelling is considered to be the most promising approach for use in population risk assessment (which is a particular focus for risk managers). For all approaches, further improvement of input data is desirable, particularly data on consumption patterns/food choices in food allergic consumers, data on minimum eliciting doses and data that can be used to evaluate whether the whole population at risk has been modelled accurately. Specific research topics were identified.

For the first time, the infrared and Raman spectra of [15N2]-succinonitrile are presented and discussed in detail. Assignments of the vibrational bands of its two rotational conformers gauche and trans, respectively, have been made for both infrared and Raman spectra. The assignments were based on a recent ab-initio force field calculation for succinonitrile, taking into account the vibrational frequencies of other succinonitrile isotopomers. There are differences in the frequencies of the vibrational bands due to the mass increase in the cyanide groups, which have been analysed in depth.

Following its release from cells during infection and inflammation, calreticulin (CRT) can act as an autoantigen in diseases such as SLE. Why CRT is a target of protective immunity and whether it may interfere with innate immunity once released from cells during inflammation is unclear. In the present study, we found that CRT was detected more frequently in SLE sera and in higher amounts than found in control sera. Approximately 40% of SLE sera tested contained autoantibodies against CRT as detected by ELISA and immunoblotting. CRT was found to be predominantly in the sera of SLE patients associated with immune complexes and C1q, and only bound to the surfaces of neutrophils in the presence of low levels of calcium and magnesium. In order to further investigate the C1q-CRT interaction, recombinant CRT and its discrete domains (N-, P-, and C-domains) were produced in Escherichia coli. CRT binds to globular head region of C1q primarily via its N- and P-domains. The N-domain was shown to be the most autoantigenic region of CRT, as the anti-CRT autoantibodies from most patients reacted against this region. CRT also altered C1q-mediated immune functions. The P-domain of CRT bound to C1q and reduced the binding of immune complexes in SLE sera to immobilized C1q. Full length CRT and its N- and P-domains were able to reduce the C1q-dependent binding of immune complexes to neutrophils and solid-phase bound C1q. We conclude that CRT, once released from leucocytes during inflammation, may not only induce an antigenic reaction, but also interfere with C1q-mediated inflammatory processes.

OBJECTIVES: To compare propofol and thiopental as anesthetic agents for electroconvulsive therapy (ECT) with respect to seizure duration, stimulus charge, clinical effect, and cognitive side effects.METHODS: Randomized, blinded study of 62 depressed patients treated with bilateral ECT. Algorithm-based charge dosing was used.RESULTS: The mean seizure duration of the patients in the thiopental group was 36.3 seconds versus 25.7 seconds in the propofol group (P = 0.001). The charge per treatment was 79.5 mC in the thiopental group versus 109.8 mC in the propofol group (P = 0.026). Sixteen patients in the propofol group (52%) reached the highest electrical dose versus 8 patients (26%) in the thiopental group (P = 0.014). No difference in response to treatment or number of treatments was observed. The mean score on Mini-Mental State Examination (MMSE) was 28.9 in the thiopental group versus 26.8 in the propofol group (P = 0.014). However, age distribution of patients completing the study differed between the groups.CONCLUSIONS: Propofol significantly decreases seizure duration without significant difference in the clinical outcome. Using the employed treatment algorithm, patients anesthetised with propofol received higher electrical charge. Mini-Mental State Examination scores suggest that this results in more severe cognitive side effects. Results, however, might be confounded by the differences in age distribution in the groups.

In 1893, Adam Politzer was the first to describe otosclerosis as a specific disease fixating the stapes. The aim of this study is to follow Politzer's research to understand how he finally explained the mechanism responsible for the fixation of the stapes. Politzer conducted his preliminary research from 1862 to 1893. From the concept of a dry catarrh of the middle ear, the fixation of the stapes became progressively associated with a specific ossification in and around the footplate. Politzer presented his first results in 1893. He completed his research by concluding in 1901 that otosclerosis had become an independent disease and should have the right to its own chapter in otologic books. He selected the word otosclerosis to describe this new pathologic entity.

Urothelial tumours which infiltrate the vesical muscle or more superficial tumours which resist localised treatment (resection+/-BCG or mytomicin C) should be considered for excision. Excision is successful in female cystectomy and in male radical cystoprostatectomy. For local tumours of the bladder (<T3), this treatment allows for a specific survival of 5 years at 90 % to be obtained. After the excision of the bladder, several types of urinary diversions may be proposed. The replacement neo-bladder by ilioplasty (orthotopic replacement) should be chosen when possible (conservation of the urether). Numerous techniques were proposed. The ilieum would appear to be the segment of the intestine best adapted for this use and of a short segment is preferable (less than 50 cm). For the creation of the neo-bladder it is necessary that the established surgical rules are followed carefully. Pre-operative preparation and post-operative treatment have become easier but must be well respected. Patients have a check-up every 6 months for the first 3 years, then annually, in order to detect possible local or secondary relapses. The correct functioning of the neo-bladder should be verified regularly in order to ensure that the bladder is emptied properly. Continence returns through auto-exercise.

Eighteen child therapy outcome studies, containing 26 treatment/control comparisons on at least one language measure, were evaluated. Four questions were addressed: (a) Does child therapy positively impact children's language proficiencies, and are its effects comparable to those obtained on nonlanguage outcome measures? (b) Do outcome effects vary across individual versus group treatment? (c) Does the type of emphasis on verbal process in treatment affect the degree of gain in the children's language proficiencies at outcome? and (d) Do effect sizes on language measures differ depending on type of presenting problem? Results indicate that child therapy has a significant positive impact on children's language proficiencies and that individual is superior to group treatment. Several trends emerged suggesting relationships between the magnitude of language change and type of emphasis on verbal processes in therapy and the childs' presenting problems. Further research on language variables is recommended.

Incidentally discovered adrenal masses are common since the advent and application of sensitive noninvasive imaging methods. The significance of these so-called "incidentalomas" and the question of further evaluation or treatment remains elusive. This report describes a retrospective study of 86 patients with incidentaloma. Adrenalectomy was performed on 26 patients during initial admission. Histologically, two cortisol-producing adenomas, an adenoma with subclinical cortisol production, and two pheochromocytomas (all of the preceding detected during the preoperative hormonal evaluation), three cystic lesions, one myelolipoma, and one hematoma were found. One primary and two metastatic adrenal carcinomas were also found in this series. Sixty patients with a nonfunctioning incidentaloma smaller than 6 cm were observed in an average of 43 months with serial CT scans performed at 3, 9, and 18 months after the initial diagnosis. Enlargement of the mass was detected in two patients; both proved to be nonfunctioning adenomas. Based on these observations, it is concluded that the initial laboratory evaluation is mandatory in cases of incidentalomas, including parameters of adrenocortical and medullar function. Hormonally active incidentalomas and those suspected for malignancy should be treated surgically. Masses greater than 6 cm should also be removed. Smaller incidentalomas without endocrine activity or signs of malignancy should be followed by CT scan at 3, 9, and 18 months after the initial diagnosis.

Quercetin, a natural constituent abundantly present in grapes, red wine, and other food products, is known to possess potent antiproliferative effects against various malignant cells. The present study aims to investigate the effect of quercetin on the apoptosis and morphology of gastric carcinoma BGC-823 cells, as well as the probable mechanism, in an effort to identify an effective drug as a potential candidate for gastric cancer. Gastric carcinoma BGC-823 cells were treated with quercetin, and cell morphology was determined by light microscopy and transmission electron microscopy. Apoptosis and cell cycle were measured by flow cytometry, using propidium iodide staining. The apoptotic protein expression of caspase-3, Bcl-2 and Bax was detected by Western blot. Quercetin induced apoptosis in BGC-823 cell. Some morphologic features of apoptosis were found, such as cell shrinkage or even apoptosis body. Quercetin changed the apoptotic protein expression. These results indicate that quercetin can induce apoptosis of the BGC-823 cells. A decrease in Bcl-2/Bax ratio with the increased expression of caspase-3 provides evidence that quercetin-induced apoptosis may be mediated via the mitochondrial pathway.

AIM: This study aimed to determine the morphological types of the foetal sigmoid colon and establish if the anatomical predisposition to sigmoid volvulus is present in Black African foetuses. Sigmoid volvulus affects Black Africans in our province more frequently than other ethnic groups, and males more than females. Its aetiology remains speculative, with factors being proposed that do not suggest the pathogenetic mechanisms or the gender bias. Previous anatomical studies have suggested that Black Africans have a predisposition to sigmoid volvulus due to an elongated sigmoid colon. We investigated whether this phenomenon occurred during foetal development.METHOD: Foetuses were donated from local hospitals. Population groups were defined as Black African and non-African. After dissection, the anatomy of the sigmoid colon was described according to level of origin, classic or elongated type and shape (broad or narrow).RESULTS: A total of 296 Black African foetuses and 37 non-African foetuses were donated. The sigmoid colon in the majority of foetuses in all gestational age groups had a low level of origin. An elongated colon was seen in 68% of Black African and 17% of non-African foetuses (P < 0.0001). Among Black Africans there was a higher proportion of elongated sigmoid colon among male (73%) than female foetuses (62%) (P = 0.044). In foetuses with an elongated sigmoid colon the broad shape was more common in females than males and the long-narrow shape was more common in males (P = 0.038).CONCLUSION: The elongated sigmoid colon seen in Black Africans is present in utero and occurs more frequently in Black African males. A narrow shape is more common in male foetuses and the broad shape is more common in female foetuses. These anatomical features may be the cause of the predisposition to sigmoid volvulus in Black African adults.

Guinea pigs were subjected to unilateral thermocoagulation of the frontal, parietal, temporal and occipital cortex, and were allowed to survive 4 and 7 days. Routine electron microscopic technique was employed to examine orthograde degenerative changes in the ipsilateral pontine nuclei. Following four days survival the degenerating corticofugal synaptic boutons (d.s.b.) exhibited features attributed to all three basic degeneration types: dark, filamentous, and light. Most of the synaptic boutons arising in the frontal, parietal and occipital cortex display filamentous degenerative changes, and measure approximately 3 micrometers. The temporopontine axons terminate as dark d.s.b. Also, some d.s.b. following frontal ablation (collaterals of the corticospinal tract?), and a small number of occipitopontine d.s.b. (visual associative cortex?), develop dark degenerative changes. Most of the dark d.s.b. measure less than 2 micrometer. The light d.s.b., with mean diameter 2 micrometer, are rarely found following frontal and occipital lesions. Following 7 days survival almost exclusively dark d.s.b. are to be observed--a great part of them, apparently, representing the late stage of the evolution of the filamentous and light degeneration. No d.s.b. were encountered in the pontine nuclei contralateral to the cortical lesion. In good agreement with preceding studies in other animal species, the present study provides a morphological evidence for a complex, multichannel relationships between the various regions of the cerebral cortex and the pontine nuclei.

114In(m)-labelled heat-damaged erythrocytes (HDRBC) in BDF1 mice were quantitatively much less successful in spleen targeting than in the rat. Unheated labelled cells (NRBC) were an even less effective vector. Radiobiological effects manifest in the spleen (weight loss) and white cell count were nevertheless substantial, comparable in both groups and unchanged after 14 days. Considerable early renal loss of indium is inferred in the NRBC animals, but in the HDRBC group a high proportion was retained in the liver. Progenitor cell marrow cultures indicated acute transitory lethal effects with only partial recovery at termination; the marrow concentration of indium was the same in both groups.

CGA-N46 is a small antifungal-derived peptide and consists of the 31st-76th amino acids of the N-terminus of human chromogranin A. Polycistronic expression of recombinant CGA-N46 in Bacillus subtilis DB1342 was used to improve its production, but the yield of CGA-N46 was still low. In the present study, response surface methodology (RSM) was used to optimize culture medium composition and growth conditions of the engineered strain B. subtilis DB1342(p-3N46) for the further increase in CGA-N46 yield. The results of two-level factorial experiments indicated that dextrin and tryptone were significant factors affecting CGA-N46 expression. Central composite design (CCD) was used to determine the ideal conditions of each significant factors. From the results of CCD, the optimal medium composition was predicted to be dextrin 16.6 g/L, tryptone 19.2 g/L, KH2PO4·H2O 6 g/L, pH 6.5. And the optimal culture process indicated inoculation of B. subtilis DB1342(p-3N46) seed culture into fresh culture medium at 5 % (v/v), followed by expression of CGA-N46 for 56 hours at 30 °C induced by 2 % (v/v) sucrose after one hour of shaking culture. To test optimal CGA-N46 peptide expression, the yeast growth inhibition assay was employed and it was found that under optimal culture conditions, CGA-N46 inhibited the growth of Candida albican by 42.17, 30.86 % more than that in the pre-optimization conditions. In summary, RSM can be used to optimize expression conditions of CGA-N46 in engineered strains B. subtilis DB1342(p-3N46).

Imprinted genes play important roles in mammalian growth, development and behavior. In this study, we obtained 1568 bp mRNA sequence of porcine DIO3 (deiodinase, iodothyronine, type III), and also identified its imprinting status during porcine fetal development. The complete open reading frame (ORF) encoding 278 amino acids. The porcine DIO3 mRNA was expressed predominantly in backfat, mildly in liver, uterus, kidney, heart, small intestine, muscle and stomach, and almost absent in spleen and lung. A single nucleotide polymorphism in exon (A/C (687)) was used to investigate the allele frequencies in different pig breeds and the imprinting status in porcine embryonic tissues. The results indicate that DIO3 was imprinted in all the tested tissues. Statistical analysis showed the DIO3 gene polymorphism was significantly associated with almost all the fat deposition and carcass traits, including lean meat percentage (LMP), fat meat percentage (FMP), ratio of lean to fat (RLF), shoulder fat thickness (SFT), sixth-seventh rib fat thickness (RFT), buttock fat thickness (BFT), loin eye area (LEA), and intramuscular fat (IMF).

Notch receptors are key regulators of nervous system development and promoters of neural stem cells renewal and proliferation. Defects in the expression of Notch genes result in severe, often lethal developmental abnormalities. Notch3 is generally thought to have a similar proliferative, anti-differentiation and gliogenic role to Notch1. However, in some cases, Notch3 has an opposite, pro-differentiation effect. Here, we show that Notch3 segregates from Notch1 and is transiently expressed in adult rat and mouse spinal cord neuron precursors and immature neurons. This suggests that during the differentiation of adult neural progenitor cells, Notch signalling may follow a modified version of the classical lateral inhibition model, involving the segregation of individual Notch receptors. Notch3 knockout mice, otherwise neurologically normal, are characterized by a reduced number of mature inhibitory interneurons and an increased number of highly excitable immature neurons in spinal cord laminae I-II. As a result, these mice have permanently lower nociceptive thresholds, similar to chronic pain. These results suggest that defective neuronal differentiation, for example as a result of reduced Notch3 expression or activation, may underlie human cases of intractable chronic pain, such as fibromyalgia and neuropathic pain.

Death rates in California for hypertension-related diseases during 1969-71 and 1979-81 are compared. During both periods, age-standardized rates for a composite hypertension-related mortality category are highest for blacks, followed by whites, and lowest for Asians and Pacific Islanders. Filipinos who have high prevalence rates of hypertension record low rates of hypertension-related mortality. After adjusting for the comparability ratio, the age-standardized hypertension-related death rate declined by more than 28 percent between 1969-71 and 1979-81. The decrease was greatest at age 15-44 years. Of all major hypertension-related diseases, cerebrovascular diseases registered consistently large percentage declines in mortality for all age and race groupings examined. Possible reasons for the considerable decline in hypertension-related mortality and low death rates for Asians and Pacific Islanders are discussed. The combined effects of improved population awareness, level of treatment, and control of hypertension; a greater knowledge of cardiovascular risk factors and associated modifications of behavior; and improved medical technology and care may have contributed to the decline.

The effect of indomethacin (1 mg/kg) on gas exchange was studied in ten patients with hypoxaemic respiratory failure precipitated by bacterial pneumonia. Mean arterial oxygen tension (PaO2) improved significantly (79 +/- 16 mm Hg to 98 +/- 20 mm Hg) but the response varied between patients: five showed substantial responses (27-42 mm Hg), three lesser responses (7-9 mm Hg), and two no response. Similar changes were found in the alveolar-arterial oxygen gradient and the ratio of PaO2 to fractional inspired oxygen concentration. In two responders studied further, PaO2 had fallen to baseline values 4-6 h later and a repeat indomethacin challenge again increased PaO2 by greater than 25 mm Hg with concomitant changes in pulmonary shunt. There were no significant changes in the other gas-exchange or haemodynamic variables measured and there was no clear reason for the variability in response to indomethacin. These results suggest a role for products of the cyclo-oxygenase pathway of arachidonic acid metabolism in the pathogenesis of hypoxaemia in patients with severe bacterial pneumonia.

BACKGROUND: Biogeographic patterns of species invasions hold important clues to solving the recalcitrant 'who', 'where', and 'why' questions of invasion biology, but the few existing studies make no attempt to distinguish alien floras (all non-native occurrences) from invasive floras (rapidly spreading species of significant management concern), nor have invasion biologists asked whether particular habitats are consistently invaded by species from particular regions.METHODOLOGY/PRINCIPAL FINDINGS: Here I describe the native floristic provenances of the 2629 alien plant taxa of the Eastern Deciduous Forest of the Eastern U.S. (EUS), and contrast these to the subset of 449 taxa that EUS management agencies have labeled 'invasive'. Although EUS alien plants come from all global floristic regions, nearly half (45%) have native ranges that include central and northern Europe or the Mediterranean (39%). In contrast, EUS invasive species are most likely to come from East Asia (29%), a pattern that is magnified when the invasive pool is restricted to species that are native to a single floristic region (25% from East Asia, compared to only 11% from northern/central Europe and 2% from the Mediterranean). Moreover, East Asian invaders are mostly woody (56%, compared to just 23% of the total alien flora) and are significantly more likely to invade intact forests and riparian areas than European species, which dominate managed or disturbed ecosystems.CONCLUSIONS/SIGNIFICANCE: These patterns suggest that the often-invoked 'imperialist dogma' view of global invasions equating invasion events with the spread of European colonialism is at best a restricted framework for invasion in disturbed ecosystems. This view must be superseded by a biogeographic invasion theory that is explicitly habitat-specific and can explain why particular world biotas tend to dominate particular environments.

The prospective study of evaluation temporomandibular dysfunction by air computerized tomography of the joint are reported. The authors propose a new injection technique of both articular compartments by CT Scan location. With thin sagittal computed tomography sections, closed mouth and open mouth position, this study is made. Preliminary data from air computerized tomography of the joint imaging show directly the disk with anterior and posterior attachments. The authors present the normal results and major pathology found in the evaluationof temporomandibular dysfunction. CT Scan puncture is more precise. The air computerized tomography imaging of the joint was undoubtedly superior to that of conventional opaque joint tomography, but dynamic notion is much less than in the absence of videoscopy.

Increased incidences of supernumerary ribs (SNR) are a relatively common finding in standard teratology bioassays, and previous studies have indicated a possible correlation between their occurrence and general maternal stress. The present study describes the effects of immobilization stress on the induction of supernumerary ribs. To isolate the sensitive period of SNR induction, Sprague-Dawley rats and CD-1 mice were treated with 300 and 1,500 mg/kg, respectively, of sodium salicylate on single days 7-11 of gestation. In the rat, day 10 was found to be the sensitive period of lumbar rib induction (32% SNR vs 10% on other days) while day 9 was critical in the mouse (71% SNR vs less than 29% on other days). In a second set of experiments, maternal stress was accomplished by restraining two groups of gravid females in the supine position for 12 hours on the predetermined sensitive day. One group was immobilized from 9 am to 9 pm, while the second group was restrained from 9 pm to 9 am. Concurrent controls were food and water deprived for similar periods. Additional untreated controls were also included. An increase in supernumerary ribs was noted in stressed mice but not in rats. The 9 am to 9 pm mouse group exhibited the highest increase in supernumerary ribs (41%) as well as significant incidences of fused ribs and exencephaly. A significant linear relationship between maternal weight loss during treatment and increases in supernumerary ribs was also noted. Results suggest that for some compounds, supernumerary ribs may be the indirect result of agent-induced, generalized maternal stress in the CD-1 mouse.

Salicylate, one of the most widely used drugs, produces at repetitive high doses reversible tinnitus and hearing loss. Neural correlates of hearing loss have long been established, whereas they remain elusive for tinnitus. The average spectrum of electrophysiological cochleoneural activity (ASECA), a measure of spontaneous auditory nerve activity, was monitored in guinea pigs over weeks of salicylate administration. Auditory nerve compound action potential (CAP) was also recorded to monitor acoustic sensitivity. In the first days of treatment, ASECA decreased acutely during hours after salicylate administration; after several days this decrease could be reduced. Over weeks of treatment the level of ASECA increased progressively. No change in CAP threshold was observed. The ASECA decrease induced by a contralateral broadband noise remained unchanged. At the end of treatment, acoustic tuning of ASECA showed a partially decreased sensitivity. After cessation of treatment the ASECA level returned progressively to initial values. In control animals delivery of an ipsilateral acoustic noise could reproduce the ASECA increase observed in long-term salicylate-treated animals. This white noise was of moderate sound pressure level and it elevated slightly CAP thresholds at high frequencies. These data provide evidence for salicylate-induced ASECA alterations without changes in CAP thresholds, in accord with clinical reports of tinnitus being the first subjective sign of salicylate ototoxicity. The similarities in occurrence, development, reversibility, frequency content, and acoustic level support the idea that ASECA changes, which indicates alterations of spontaneous eighth nerve activity and reflects the presence of salicylate-induced high-pitch tinnitus.

We and others have previously documented increased resting and exercise-induced skeletal muscle blood flow in young subjects with Type I (insulin-dependent) diabetes mellitus compared with healthy controls. Both NO and prostanoids are important regulators of vascular tone and may therefore contribute to this hyperaemia. The aim of the present study was to determine the contribution of NO and vasodilator prostanoids to this skeletal muscle hyperaemia in diabetes. We assessed the effects of infusion into the intrabrachial artery of the cyclo-oxygenase inhibitor acetylsalicylic acid (ASA; aspirin) and of the L-arginine analogue N(G)-monomethyl-L-arginine (L-NMMA) on skeLetal muscle blood flow in subjects with Type I diabetes mellitus (DM subjects) and control subjects. Blood flow was measured by venous occlusion plethysmography. Isotonic forearm exercise involved 2 min of wrist flexion and extension. Resting flow (forearm blood flow; FBF) was augmented in DM subjects, as was peak exercise-related blood flow (PFBF) and the volume repaid to the forearm 5 min after exercise (AUC 5, where AUC is area under the flow-time curve) (P<0.05), even when accounting for differences in basal flow. Infusion of L-NMMA reduced resting flow by 48% in controls (P<0.005) and by 12% in DM subjects (not significant). L-NMMA reduced PFBF and AUC 5 by 29% (P<0.05) and 39% (P<0.0005) respectively in controls, but had no significant effect on these parameters in DM subjects. Infusion of ASA reduced FBF, PFBF and AUC 5 in both DM (P<0.05) and control (P<0.05) subjects, but the magnitude of this reduction was greater in DM than in control subjects (ANOVA, P<0.05), even when differences in resting FBF were accounted for. Indeed, ASA eliminated the differences in FBF, PFBF and AUC 5 between DM and control subjects. Thus increased release of vasodilator prostanoids, rather than of NO, appears to account for skeletal muscle hyperaemia in Type I diabetes.

Kinetics of fosfomycin were investigated in six patients undergoing continuous ambulatory peritoneal dialysis. Each subject received both an i.v. and an i.p. 1 g dose of fosfomycin with a one week washout between doses. Fosfomycin was assayed by a microbiological diffusion technique. After intravenous injection the fosfomycin serum kinetic parameters were as followed: elimination half-life (t1/2 beta) 38.4 +/- 8.7 h; volume of distribution 0.32 +/- 0.02 l/kg; total plasma clearance 7.0 +/- 1.4 ml/min and peritoneal clearance 3.2 +/- 0.2 ml/min. Dialyzate fosfomycin concentrations reached a maximum mean value of 32.2 +/- 2.8 micrograms/ml at 4 h post-injection and fosfomycin was detectable in dialyzate samples for up to 72 hours post-dosing. After intraperitoneal instillation, fosfomycin appeared in the serum rapidly and the mean peak plasma concentration was 36.2 +/- 2.8 micrograms/ml at the 4th h. The absorption rate (ka) was 0.580 +/- 0.039 h-1 and the absorption of fosfomycin from peritoneal space was 68.4 +/- 6.0%. These data suggest a bidirectional exchange through the peritoneal membrane. Intraperitoneal administration of 1 g either 48 h apart for anephric patients or 36 h apart for patients with residual renal function may achieve therapeutic serum concentrations.

MiR-34c is considered a potent tumour suppressor because of its negative regulation of multiple target mRNAs that are critically associated with tumorigenesis and metastasis. In the present study, we demonstrated a novel target of miR-34c, KITLG, which has been implicated in colorectal cancer (CRC). First, we found a significant negative relationship between miR-34c and KITLG mRNA expression levels in CRC cell lines, including HT-29, HCT-116, SW480 and SW620 CRC cell lines. In silico analysis predicted putative binding sites for miR-34c in the 3' untranslated region (3'UTR) of KITLG mRNA. A dual-luciferase reporter assay further confirmed that KITLG is a direct target of miR-34c. Then, the cell lines were infected with lentiviruses expressing miR-34c or a miR-34c specific inhibitor. Restoration of miR-34c dramatically reduced the expression of KITLG mRNA and protein, while silencing of endogenous miR-34c increased the expression of KITLG protein. The miR-34c-mediated down-regulation of KITLG was associated with the suppression on proliferation, cellular transformation, migration and invasion of CRC cells, as well as the promotion on apoptosis. Knockdown of KITLG by its specific siRNA confirmed a critical role of KITLG down-regulation for the tumour-suppressive effects of miR-34c in CRC cells. In conclusion, our results demonstrated that miR-34c might interfere with KITLG-related CRC and could be a novel molecular target for CRC patients.

BACKGROUND: Endo-1,4-â-mannanase is an enzyme that can catalyze the random hydrolysis of â-1, 4-mannosidic linkages in the main chain of mannans, glucomannans and galactomannans and has a number of applications in different biotechnology industries. Penicillium oxalicum is a powerful hemicellulase-producing fungus (Bioresour Technol 123:117-124, 2012); however, few previous studies have focused on the cloning and expression of the endo-1,4-â-mannanase gene from Penicillium oxalicum.RESULTS: A gene encoding an acidophilic thermostable endo-1,4-â-mannanase (E.C. 3.2.1.78) from Penicillium oxalicum GZ-2, which belongs to glycoside hydrolase family 5, was cloned and successfully expressed in Pichia pastoris GS115. A high enzyme activity (84.4 U mL(-1)) was detected in the culture supernatant. The recombinant endo-1,4-â-mannanase (rPoMan5A) was tagged with 6 ? His at its C-terminus and purified using a Ni-NTA Sepharose column to apparent homogeneity. The purified rPoMan5A showed a single band on SDS-PAGE with a molecular mass of approximately 61.6 kDa. The specific activity of the purified rPoMan5A was 420.9 U mg(-1) using locust bean gum as substrate. The optimal catalytic temperature (10 min assay) and pH value for rPoMan5A are 80 °C and pH 4.0, respectively. The rPoMan5A is highly thermostable with a half-life of approximately 58 h at 60 °C at pH 4.0. The K m and V max values for locust bean gum, konjac mannan, and guar gum are 7.6 mg mL(-1) and 1425.5 ìmol min(-1) mg(-1), 2.1 mg mL(-1) and 154.8 ìmol min(-1) mg(-1), and 2.3 mg mL(-1) and 18.9 ìmol min(-1) mg(-1), respectively. The enzymatic activity of rPoMan5A was not significantly affected by an array of metal ions, but was inhibited by Fe(3+) and Hg(2+). Analytical results of hydrolytic products showed that rPoMan5A could hydrolyze various types of mannan polymers and released various mannose and manno-oligosaccharides, with the main products being mannobiose, mannotriose, and mannopentaose.CONCLUSION: Our study demonstrated that the high-efficient expression and secretion of acid stable and thermostable recombinant endo-1, 4-â-mannanase in Pichia pastoris is suitable for various biotechnology applications.

The characterization of phosphorylation state(s) of a protein is best accomplished by using isolated or enriched phosphoprotein samples or their corresponding phosphopeptides. The process is typically time-consuming as, often, a combination of analytical approaches must be used. To facilitate throughput in the study of phosphoproteins, a microreactor that enables a novel strategy for performing fast proteolytic digestion and selective phosphopeptide enrichment was developed. The microreactor was fabricated using 100 ìm i.d. fused-silica capillaries packed with 1-2 mm beds of C18 and/or TiO2 particles. Proteolytic digestion-only, phosphopeptide enrichment-only, and sequential proteolytic digestion/phosphopeptide enrichment microreactors were developed and tested with standard protein mixtures. The protein samples were adsorbed on the C18 particles, quickly digested with a proteolytic enzyme infused over the adsorbed proteins, and further eluted onto the TiO2 microreactor for enrichment in phosphopeptides. A number of parameters were optimized to speed up the digestion and enrichments processes, including microreactor dimensions, sample concentrations, digestion time, flow rates, buffer compositions, and pH. The effective time for the steps of proteolytic digestion and enrichment was less than 5 min. For simple samples, such as standard protein mixtures, this approach provided equivalent or better results than conventional bench-top methods, in terms of both enzymatic digestion and selectivity. Analysis times and reagent costs were reduced ~10- to 15-fold. Preliminary analysis of cell extracts and recombinant proteins indicated the feasibility of integration of these microreactors in more advanced workflows amenable for handling real-world biological samples. Graphical Abstract ?.

Fever is associated with poor outcome in acute stroke. Forty-two consecutive, normothermic patients with acute ischemic stroke were, within 24 hours from symptom onset, randomized to either receive 4 g acetaminophen daily (n = 20) or matched placebo (n = 22). Fever of greater than 37.5 degrees C occurred in 36.4% of patients in the placebo group, compared with 5.0% in the acetaminophen group (Fisher's exact test, p = 0.014). Prophylactic antipyretic treatment with acetaminophen may be effective in the prevention of fever after acute ischemic stroke.

Human toxocarosis is one of the most widespread and prevalent helminthic zoonosis in many countries, including Slovakia. The aim was to evaluate the usefulness of IgA anti-Toxocara antibody detection in the serodiagnosis of toxocarosis. The levels of specific IgA antibodies were determined by excretory-secretory (ES)-enzyme-linked immunosorbent assay (ELISA). The IgA seropositivity in IgG anti-Toxocara seropositive patients (n = 52) was 32.7% and found to be highest in the oldest age groups (P = 0.026). The presence of IgA in suspected patients for toxocarosis were evaluated in respect to some characteristics of examined persons. Substantially higher IgA seropositivity was detected in patients with increased total IgE (44.8%) than in subjects with a normal level of IgE (17.4%; P = 0.036). No associations (P > 0.05) were found between IgA seropositivity and sex, level of specific IgG antibodies, avidity of IgG, eosinophilia, domicile, geophagia, traveling abroad, dog/cat ownership, or clinical symptoms. The IgA-ELISA showed sensitivity of 57.1% and specificity of 100%. Mild correlations (r = 0.302, r = 0.305, r = - 0.409) were observed between the levels of anti-Toxocara IgA antibodies and age, the amounts of eosinophils and IgA antibody levels, the amounts of eosinophils, and the values of IgG avidity, respectively. The presence of anti-Toxocara IgA may facilitate the diagnosis of toxocarosis and may well be useful for the determination of acute Toxocara infection. Moreover, this test should be accompanied by other immunological markers of examined patients (e.g., increased total IgE, eosinophilia, and low-avidity IgG antibodies).

Accumulating evidence has suggested that cellular production of superoxide acts as an intracellular messenger to regulate gene expression and modulate cellular activities. In this report, we set out to investigate the role of active H-ras-mediated superoxide production on tumor cell malignancy in a SV-40 transformed human lung WI-38 VA-13 cell line. Stable transfection and expression of constitutively active mutant V12-H-ras (V12-H-ras) dramatically increased intracellular production of superoxide. The expression of V12-H-ras significantly enhanced cell proliferation, migration and resistance to TNF-alpha treatment compared to that of parental and vector control cells, while expression of wild type H-ras (WT-H-ras) only had modest effects. Upon scavenging by superoxide dismutase and other molecules that decrease the intracellular level of active H-ras mediated superoxide production, cell proliferation, migration and resistance to TNF-alpha were significantly reduced. Furthermore, we demonstrated that the activation of membrane NADPH oxidase activity by expression of active H-ras contributed to the intracellular superoxide production. The causal relationship between membrane superoxide production and increased cell proliferation, migration, and resistance to TNF-alpha by the expression of active H-ras, has provided direct evidence to demonstrate that superoxide acts as an intracellular messenger to cascade ras oncogenic signal relay and to modulate tumor malignant activity.

Purpose: To assess tracheobronchial angles and their changes on combined inspiratory and expiratory thoracic computed tomography (CT) scans and to determine correlations between tracheobronchial angles and several indices of chronic obstructive pulmonary disease (COPD).Materials and methods: A total of 80 smokers underwent combined inspiratory and expiratory CT scans. Of these, 65 subjects also performed spirometry and 55 patients were diagnosed with COPD. On CT scans, 3-dimensinal tracheobronchial angles (trachea-right main bronchus [RMB], trachea-left main bronchus [LMB], and RMB-LMB) were automatically measured by software. Lung volumes at inspiration and expiration were also automatically calculated. Changes in tracheobronchial angles between inspiration and expiration were assessed by the Mann-Whitney test. Correlations of the angles with lung volume, airflow limitation, and CT-based emphysema index were evaluated by Spearman rank correlation.Results: The trachea-LMB angle was significantly smaller and the RMB-LMB angle was significantly larger at expiration than inspiration (P<0.0001). The trachea-LMB and RMB-LMB angles were significantly correlated with lung volume, particularly at expiration. The RMB-LMB angle was significantly correlated with airflow limitation and CT emphysema index (P<0.001-0.05) at inspiration and expiration, suggesting that narrowed RMB-LMB angle indicates more severe airflow limitation and larger extent of emphysema.Conclusion: Tracheobronchial angles change during respiration and are correlated with severity of COPD and emphysema.

The organization and control of stem cells is a key issue in epithelial cell biology. The small intestinal murine crypt is a useful tissue to study such problems since stem cells are known to be located at specific positions at the bottom of the crypt where they are self maintaining. Recent data suggest, that (1) the number of active stem cells in a crypt can fluctuate, (2) the immediate progeny of a single stem cell can replace other stem cells eventually leading to monoclonality and (3) the life cycle of crypts may be linked to stem cell dynamics. It is the objective of this paper to suggest a stochastic state-dependent model of stem cell and crypt growth which can explain and-link these phenomena into one comprehensive framework. Monte Carlo simulations are performed to show consistently with available data. The model explains the size distribution of small intestinal crypts in steady state, the observations of stem cell fluctuations and monoclonality conversion, recovery of the crypt population after moderate damage and the rate of crypt fission and extinction. The key assumption of this model is an autoregulatory control of stem cell growth.

OBJECTIVES: We sought to identify a novel gene for dilated cardiomyopathy (DCM).BACKGROUND: DCM is a heritable, genetically heterogeneous disorder that remains idiopathic in the majority of patients. Familial cases provide an opportunity to discover unsuspected molecular bases of DCM, enabling pre-clinical risk detection.METHODS: Two large families with autosomal-dominant DCM were studied. Genome-wide linkage analysis was used to identify a disease locus, followed by fine mapping and positional candidate gene sequencing. Mutation scanning was then performed in 278 unrelated subjects with idiopathic DCM, prospectively identified at the Mayo Clinic.RESULTS: Overlapping loci for DCM were independently mapped to chromosome 10q25-q26. Deoxyribonucleic acid sequencing of affected individuals in each family revealed distinct heterozygous missense mutations in exon 9 of RBM20, encoding ribonucleic acid (RNA) binding motif protein 20. Comprehensive coding sequence analyses identified missense mutations clustered within this same exon in 6 additional DCM families. Mutations segregated with DCM (peak composite logarithm of the odds score >11.49), were absent in 480 control samples, and altered residues within a highly conserved arginine/serine (RS)-rich region. Expression of RBM20 messenger RNA was confirmed in human heart tissue.CONCLUSIONS: Our findings establish RBM20 as a DCM gene and reveal a mutation hotspot in the RS domain. RBM20 is preferentially expressed in the heart and encodes motifs prototypical of spliceosome proteins that regulate alternative pre-messenger RNA splicing, thus implicating a functionally distinct gene in human cardiomyopathy. RBM20 mutations are associated with young age at diagnosis, end-stage heart failure, and high mortality.

Research suggests that select processing speed measures can also serve as embedded validity indicators (EVIs). The present study examined the diagnostic utility of Wechsler Adult Intelligence Scale-Fourth Edition (WAIS-IV) subtests as EVIs in a mixed clinical sample of 205 patients medically referred for neuropsychological assessment (53.3% female, mean age = 45.1). Classification accuracy was calculated against 3 composite measures of performance validity as criterion variables. A PSI ?79 produced a good combination of sensitivity (.23-.56) and specificity (.92-.98). A Coding scaled score ?5 resulted in good specificity (.94-1.00), but low and variable sensitivity (.04-.28). A Symbol Search scaled score ?6 achieved a good balance between sensitivity (.38-.64) and specificity (.88-.93). A Coding-Symbol Search scaled score difference ?5 produced adequate specificity (.89-.91) but consistently low sensitivity (.08-.12). A 2-tailed cutoff on the Coding/Symbol Search raw score ratio (?1.41 or ?3.57) produced acceptable specificity (.87-.93), but low sensitivity (.15-.24). Failing ?2 of these EVIs produced variable specificity (.81-.93) and sensitivity (.31-.59). Failing ?3 of these EVIs stabilized specificity (.89-.94) at a small cost to sensitivity (.23-.53). Results suggest that processing speed based EVIs have the potential to provide a cost-effective and expedient method for evaluating the validity of cognitive data. Given their generally low and variable sensitivity, however, they should not be used in isolation to determine the credibility of a given response set. They also produced unacceptably high rates of false positive errors in patients with moderate-to-severe head injury. Combining evidence from multiple EVIs has the potential to improve overall classification accuracy. (PsycINFO Database Record

Dichotic listening scores and information on lesion size and location were obtained for aphasic patients. All patients had a reduction in dichotic scores at 1 month postonset of symptoms, but some patients returned to normal levels of performance by 6 months postonset. Lesion location was the main determinant of patterns of performance. Right ear scores from the dichotic digit test closely reflected the presence or absence of lesions involving Heschl's gyrus. What appeared to be an effect of lesion volume was confounded by the relationship between increasing lesion volume and a greater likelihood of damage to Heschl's gyrus.

The polymerase chain reaction (PCR) method was used to characterize trypanosome infections in tsetse flies from 3 sleeping sickness foci in Cameroon. The predominant tsetse species found was Glossina palpalis palpalis. An average infection rate of 12.1% was revealed by microscopical examination of 888 non-teneral tsets flies. PCR amplification analyses for trypanosome identification were carried out on 467 flies, with primer sets specific for Trypanosoma (Trypanozoon) brucei s.1., T. (Duttonella) vivax, T. (Nannomonas) simiae and forest type T. (Nannomonas) congolense. Of 467 flies 93 were positive by microscopical analysis while PCR succeeded in identifying 89 positive flies. Of the PCR-positive flies 34 (38.2%) were negative by microscopical examination. PCR amplification, when compared to the parasitological technique, gave a higher estimate of infection rate of trypanosomes in natural tsetse populations. The PCR technique did, however, fail to identify 40.9% (38/93) of the parasitologically positive flies. The reasons for this failure are discussed. The overall prevalence of mixed infections, assessed by PCR, was 37.1%; the majority (72.7%) involved T. brucei and forest type T. congolense.

OBJECTIVES: To assess the association between neighbourhood environmental variables and excessive transport-related and leisure-time sedentary behaviour (ETSB and ELSB, respectively) amongst adult women in Southern Brazil.METHODS: A cross-sectional survey was conducted using a representative sample of 1079 women 20 to 69 years of age who lived in 44 neighbourhoods. Each neighbourhood was determined by drawing a 400-m buffer around the median point of the participants' homes. Neighbourhoods were assessed by audit and by using government data. Sedentary behaviour (SB) and the remaining individual variables were assessed via questionnaire that was administered as an interview. Multilevel logistic regression analysis was conducted.RESULTS: In the adjusted analysis, women who lived in neighbourhoods with a higher percentage of terrain slope and fewer public recreation areas per resident had significantly (p ? 0.05) increased odds of exhibiting ETSB and ELSB, with 50% and 40% increases, respectively.CONCLUSIONS: The results show that some aspects of the neighbourhood environment such as terrain slope and total public recreation areas per resident are related to specific SBs, indicating that improvements in neighbourhoods can reduce SB in women.

Underlying cardiovascular disease (CVD) is a risk factor for the exacerbation of air pollution health effects. Pulmonary oxidative stress, inflammation, and altered iron (Fe) homeostasis secondary to CVD may influence mammalian susceptibility to air pollutants. Rodent models of CVD are increasingly used to examine mechanisms of variation in susceptibility. Baseline cardiac and pulmonary disease was characterized in healthy normotensive Wistar Kyoto (WKY) rats, cardiovascular compromised spontaneously hypertensive rats (SHR), and spontaneously hypertensive heart failure (SHHF) rats. Blood pressure, heart rate, and breathing frequencies were measured in rats 11 to 12 wk of age, followed by necropsy at 14 to 15 wk of age. Blood pressure and heart rate were increased in SHR and SHHF relative to WKY rats (SHR > SHHF > WKY). Increased breathing frequency in SHHF and SHR (SHR > SHHF > WKY) resulted in greater minute volume relative to WKY. Bronchoalveolar lavage fluid (BALF) protein and neutrophils were higher in SHHF and SHR relative to WKY (SHHF >> SHR > WKY). Lung ascorbate and glutathione levels were low in SHHF rats. BALF Fe-binding capacity was decreased in SHHF relative to WKY rats and was associated with increased transferrin (Trf) and ferritin. However, lung ferritin was lower and Trf was higher in SHHF relative to WKY or SHR rats. mRNA for markers of inflammation and oxidative stress (macrophage inflammatory protein [MIP]-2, interleukin [IL]-1alpha, and heme oxygenase [HO]-1) were greater in SHHF and SHR relative to WKY rats. Trf mRNA rose in SHR but not SHHF relative to WKY rats, whereas transferrin receptors 1 and 2 mRNA was lower in SHHF rats. Four of 12 WKY rats exhibited cardiac hypertrophy despite normal blood pressure, while demonstrating some of the pulmonary complications noted earlier. This study demonstrates that SHHF rats display greater underlying pulmonary complications such as oxidative stress, inflammation, and impaired Fe homeostasis than WKY or SHR rats, which may play a role in SHHF rats' increased susceptibility to air pollution.

We previously described the differential distribution of major gangliosides (GM1, GD1a, GD1b, GT1b and GQ1b) in adult rat brain detected by specific antibodies (Kotani, M., Kawashima, I., Ozawa, I., Terashima, T. and Tai, T. Glycobiology, 3, 137-146, 1993). We report here the distribution of minor gangliosides in the adult rat brain by an immunofluorescence technique with mouse monoclonal antibodies (MAbs). Ten MAbs (GMR6, GMB28, GMR11, GMR19, GMR2, GMR7, GGR51, AMR10, NGR54 and NGR53) that specifically recognize GM3, GM2, GT1a, GD3, O-Acdisialoganglioside, GD2, GM1b, GM4, IV3NeuAc alpha-nLc4Cer and IV6NeuAc alpha-nLc4Cer, respectively, were used. Our study revealed that there is a cell type-specific expression of minor gangliosides, as well as major gangliosides, in the rat brain. In the cerebellar cortex, GM3 was expressed intensely in the white matter and slightly in the granular layer. GD3 was present in both the granular layer and the white matter, but not in the Purkinje cell layer or in the molecular layer. An O-Ac-disialoganglioside, which was suggested to be O-Ac-LD1, was detected exclusively in both the molecular layer and Purkinje cell layer. The presence of GD2 was restricted to the granular layer. GM4 was associated with some astrocytes, but not with myelin or oligodendrocytes. GM2, GT1a, GM1b, IV3NeuAc alpha-nLc4Cer and IV6NeuAc alpha-nLc4Cer gangliosides were not clearly detected in the cerebellar cortex. In other regions, such as cerebral cortex, hippocampal formation and spinal cord, the expression of the gangliosides was also highly localized to a specific cell type and layer.

BACKGROUND: Fingerprints studied by dermatoglyphics are unique for a given individual. It depends on the genetic makeup of an individual. Hypertension, a harbinger of many complications, is determined by genetic and environmental factors. In this observational study, we tried to find an association of palmar dermatoglyphic parameters and hypertension.METHOD: Two hundred fifty known hypertensives as cases and 250 normotensives as controls were enrolled after considering inclusion and exclusion criteria. Dermatoglyphic patterns on tips of fingers obtained by digital imaging were noted in both the groups, and "atd" angle was calculated using "screen protractor" software. Collected data were statistically analyzed to find any association between dermatoglyphic qualitative and dermatoglyphic quantitative patterns and hypertension.RESULT: Mean "atd" angle was higher in cases than in controls. Comparison of dermatoglyphic patterns in both the groups in various ways-both hands together, the right hand and left hand separately, similar fingers on right and left hand together, and similar fingers separately-was performed which revealed that at every level, whorls were more frequent in cases than in controls and that distribution of dermatoglyphic patterns were statistically significant in cases than in controls.CONCLUSION: Fingerprint patterns can be reliably used to identify individuals likely at risk for hypertension, and accordingly, preventive measures can be targeted. This subject area demands a need for further research and analysis with large sample size to allow dermatoglyphics to evolve into a cost-effective and handy tool for identifying individuals at risk of hypertension.

Bicuspid aortic valve (BAV) is the most common congenital heart defect and can lead to severe complications. Many studies have been conducted to evaluate the potential of geometric and hemodynamic biomarkers to predict BAV-related aortopathy. This work proposes a novel semi-automatic protocol to characterize geometry and hemodynamics of aorta and aortic valve based on 2D cine and 4D flow MRI data. The protocol was applied to 12 BAV and 12 control subjects. Statistical differences between the groups were identified and linear regression models were investigated to elucidate the potential of this protocol to investigate the risk of disease progression. Statistical differences between the groups were found for orifice eccentricity, aorta diameter, velocity, jet angle and flow displacement. Key findings of the regression analysis include the association in the proximal ascending aorta between aorta diameter and mean velocity (negative), jet angle (positive) and flow displacement (positive), in the combined cohort. Positive association between flow displacement in the proximal aorta and orifice jet angle in TAV subjects was found. This study represents a pilot application of a protocol for standardized assessment of aortic geometry and hemodynamics associated with BAV disease. Its limited need for user input is advantageous for large cohort studies.

BACKGROUND: Around 50% of cutaneous melanomas harbor the BRAF(V600E) mutation and can be treated with BRAF inhibitors. DNA carrying this mutation can be released into circulation as cell-free BRAF(V600E) (cfBRAF(V600E)). Droplet digital PCR (ddPCR) is an analytically sensitive technique for quantifying small concentrations of DNA. We studied the plasma concentrations of cfBRAF(V600E) by ddPCR in patients with melanoma during therapy with BRAF inhibitors.METHODS: Plasma concentrations of cfBRAF(V600E) were measured in 8 controls and 20 patients with advanced melanoma having the BRAF(V600E) mutation during treatment with BRAF inhibitors at baseline, first month, best response, and progression.RESULTS: The BRAF(V600E) mutation was detected by ddPCR even at a fractional abundance of 0.005% in the wild-type gene. Agreement between tumor tissue BRAF(V600E) and plasma cfBRAF(V600E) was 84.3%. Baseline cfBRAF(V600E) correlated with tumor burden (r = 0.742, P < 0.001). cfBRAF(V600E) concentrations decreased significantly at the first month of therapy (basal median, 216 copies/mL; Q1-Q3, 27-647 copies/mL; first response median, 0 copies/mL; Q1-Q3, 0-49 copies/mL; P < 0.01) and at the moment of best response (median, 0 copies/mL; Q1-Q3, 0-33 copies/mL; P < 0.01). At progression, there was a significant increase in the concentration of cfBRAF(V600E) compared with best response (median, 115 copies/mL; Q1-Q3, 3-707 copies/mL; P = 0.013). Lower concentrations of basal cfBRAF(V600E) were significantly associated with longer overall survival and progression-free survival (27.7 months and 9 months, respectively) than higher basal concentrations (8.6 months and 3 months, P < 0.001 and P = 0.024, respectively).CONCLUSIONS: cfBRAF(V600E) quantification in plasma by ddPCR is useful as a follow-up to treatment response in patients with advanced melanoma.

It has been reported that the adrenal gland is essential to the development of obesity if Ay/a Yellow obese mice (Hausberger and Hausberger 1960). Since the actual body composition data to support this report has not been published, we attempted to duplicate this observation by adrenalectomy of the Yellow mice before the onset of obesity. Two groups of Yellow mice (Ay/a) and normal mice (a/a) were either sham operated or adrenalectomized at two months of age and at four months of age. Body weight was monitored until body gain had stopped. At that time the animals were sacrified and checked for completeness of adrenal removal. Body composition of dry matter, fat, protein, and ash was determined. Adrenalectomy caused a reduction of body fat of 33% and 30% in both the a/a Control and the Ay/a Yellow mice, respectively. The adrenalectomized Yellow mice were still fatter than adrenalectomized Controls (25.3% fat vs. 10.6% fat). Yellow and control adrenalectomized mice showed similar depression in growth rate. These data suggest that while the adrenal gland is essential for complete expression of the genetic potential for fat deposition, it may not be necessary for partial expression indicating a secondary role in the development of obesity in the Ay/a Yellow mouse.

OBJECTIVE: In the United Kingdom in 2006, 5,325 persons were diagnosed with oral cancer; and in 2007 it caused around 1,850 deaths. The purpose of this study was to assess the patient awareness, in a dental access centre, of a poster and leaflet campaign providing information about smoking and excess alcohol consumption as risk factors in the development of oral cancer, and to explore dental patients' beliefs and perceptions about these risk factors.METHODS: Posters and leaflets providing information about risk factors for oral cancer were displayed in the patient waiting areas of a dental access centre. Data were collected prospectively in relation to the smoking and drinking habits of patients attending the centre. This information was used to categorise patients into one of four groups ranging from low to high consumption. During triage, patients were asked if they had read any of the information about oral cancer that was on display, and patients in the high risk groups were asked to participate in a semi-structured interview that would explore their knowledge about risk factors and their views on the delivery of healthcare messages in relation to oral cancer.RESULTS: Data on risk status and exposure to the poster and leaflet campaign were collected for 1,161 patients attending during the study period. More than 50% of these patients were smokers, with 36% in the high or very high tobacco and alcohol use groups. Approximately 40% of patients within each consumption group had read any of the information available. Nine patients agreed to be interviewed and overall knowledge about risk factors for oral cancer, even after reading the information was poor.CONCLUSION: Dental access centres attract a significant number of patients with lifestyle habits that make them vulnerable to oral cancer, and as such are well placed to deliver oral health messages to this high risk group. However, the delivery of information through a simple poster and leaflet campaign is likely to have limited impact.

PURPOSE: To assess the course of the stability and the failure rate of dental implants placed in the partially edentulous maxillae of minipigs.MATERIALS AND METHODS: Three months after tooth removal, implants were placed in 9 minipigs. Six implants (XiVE; Friadent, Mannheim, Germany) were placed on each side of the posterior maxilla after preparation of the implant sites either by an osteotome technique or with spiral drills. Implant stability was assessed by resonance frequency analysis (RFA) at the time of placement, at second-stage surgery (which took place after a healing periods of 1, 2, 3, 4, or 5 months), and after a loading period of 6 months.RESULTS: Implant stability was significantly influenced by the healing period (P = .007). Implant stability decreased after 1 to 3 months of healing for both of the placement techniques and increased after a healing period of 4 months. After implant site preparation by an osteotome technique, 6 of 12 immediately loaded implants, 18 of 24 implants loaded after healing periods of 1 to 3 months, and 1 of 18 implants loaded after a healing period of 4 or 5 months were lost. After implant site preparation using spiral drills, 7 of 12 immediately loaded implants, 12 of 24 implants loaded after healing periods of 1 to 3 months, and 2 of 18 implants loaded after healing periods of 4 or 5 months were lost. Broad overlapping of confidence intervals for the number of implant failures revealed that there was no relevant difference between immediate and early functional loading for either of the 2 techniques.DISCUSSION AND CONCLUSION: Implant loading after healing periods of 1 to 3 months did not improve implant survival compared to immediate loading in the posterior maxillae of minipigs. Not until a healing period of 4 months was reached did implant stability begin to increase. Only when functional loading was started at this point in time was maximal implant survival achieved.

INTRODUCTION: In this study, we aimed to assess the patient and treatment factors that influence the success rate and the duration of the orthodontic-surgical modality for impacted central incisors.METHODS: The records of 60 consecutively treated patients (64 impacted incisors) were retrospectively evaluated. The success rate and the duration of each stage of treatment were examined in relation to age, sex, etiology of impaction, location of the impacted tooth, and type of surgical exposure performed. Logistic regression analyses were applied.RESULTS: The group consisted of 26 male and 34 female subjects, 7.0 to 21.9 years old; 27 patients had impactions because of root dilaceration, 29 had impactions because of obstruction, and 4 had impactions with unknown causes. The overall success rate was 90.0%. Five of the 6 failures were dilacerated incisors. The average duration of treatment was 21.6 ± 8.7 months. The only factor that significantly increased the duration was the height of the impacted tooth. Dilaceration was related to a longer stage of traction and, in older patients (late mixed and full permanent dentition), to a longer finishing stage.CONCLUSIONS: The orthodontic-surgical treatment of impacted incisors is generally successful, but relatively long. Patients and parents should be warned of the risk of failure and the increased treatment duration, especially for dilacerated incisors impacted high in the alveolus.

Determinations were made of glyoxalase I and glyoxalase II acitivity in the liver of mice (BDF1 and DBA2 strains) bearing sarcoma 180 and L1210 leukemia in ascites form. A progressive decrease in the both glyoxalase I and glyoxalase II activities to about 40--60% of that of the control groups was observed within the developing period 8--9 days. Test results are interpreted in the light of the postulated role of this enzyme system in cell division and in the tumor development process.

In this study we provide evidence that distinct DNA sequences within the 5'-flanking regions of the genes for interleukin-2 (IL-2) and its receptor (IL-2R) are involved in human T-cell-specific activation of transcription by p40x, a product of human T cell leukemia virus type I (HTLV-1). The same DNA sequences appear to be responsible for induction of the genes in a T cell line, Jurkat, by mitogens. Although the IL-2 gene sequences are activated by p40x with much lower efficiency than the IL-2R gene sequences, they are synergistically activated by the p40x expression and subsequent extracellular stimulation by Concanavalin-A or anti-T3. We propose a model for two-step activation of the IL-2 autocrine loop in ATL development.

We studied the effect of 3 weeks treatment with the selective serotonin re-uptake inhibitor (SSRI), paroxetine (30 mg daily), on the neuroendocrine and hyperthermic responses to the 5-HT2C receptor agonist, m-chlorophenylpiperazine (mCPP) (0.05 mg/kg i.v.), in seven healthy volunteers. Following paroxetine treatment, both the prolactin and hyperthermic responses to mCPP were significantly attenuated. These data are consistent with experimental animal studies indicating that repeated SSRI treatment leads to a functional desensitisation of 5-HT2C receptors. This effect may be linked to the anxiolytic properties of SSRIs.

BACKGROUND: Satisfaction is important in dental care because satisfaction with care alleviates dental anxiety, influences patients' compliance and is an important indicator of quality of care.OBJECTIVES: This study was designed to determine the factors that contribute to satisfaction with dental care among patients attending the Lagos State University (LASUTH) Dental Clinic.METHODS: Across-sectional, descriptive questionnaire-based survey was conducted among adult patients attending the LASUTH Dental Clinic. The questionnaire, a modification of the Dental Satisfaction Questionnaire (DSQ), contained 19 items on a Likert-pattern scale with scores ranging from 0 to 4.RESULTS: The scores obtained for satisfaction with the dental services ranged from 19 to 75 with a mean of 55.30 +/- 11.55. The majority of respondents (305 or 87.4%) were satisfied with the services received. The items generating the highest and lowest mean satisfaction score were cleanliness/comfort of the facility and cost of services respectively. Long waiting time was the item respondents liked least about the services. There was a statistically significant relationship between the items assessing communication and respondent's gender (p = 0.001). The relationship between the overall satisfaction score and gender (p = 0.233), age category (p = 0.842) and educational status (p = 0.565) were not statistically significant.CONCLUSION: The results indicate a high level of satisfaction with services provided at the LASUTH Dental Clinic. However, there is need for improvement in communication with patients and reduction in waiting time.

The complete primary structure of the ADP/ATP carrier from beef heart mitochondria is described. Cyanogen bromide cleaves the protein into a long, N-terminally blocked fragment with Mr 22,000 (CB1) and several small peptides. The primary information was derived from liquid-phase sequencing of tryptic peptides obtained from the maleylated carrier protein and the citraconylated CB1 fragment, as well as from cleavage products with Staphylococcus aureus protease. The multitude of thermolysinolytic, tryptic, chymotryptic and peptic peptides was sequenced by manual methods. They rendered overlaps and further supported already known partial sequences. Also, the bridge between the published acidolytic C-terminal fragment A2 was thus obtained.

Research on plasticity markers in the cerebral cortex has largely focused on their timing of expression and role in shaping circuits during critical and normal periods. By contrast, little attention has been focused on the spatial dimension of plasticity-stability across cortical areas. The rationale for this analysis is based on the systematic variation in cortical structure that parallels functional specialization and raises the possibility of varying levels of plasticity. Here, we investigated in adult rhesus monkeys the expression of markers related to synaptic plasticity or stability in prefrontal limbic and eulaminate areas that vary in laminar structure. Our findings revealed that limbic areas are impoverished in three markers of stability: intracortical myelin, the lectin Wisteria floribunda agglutinin, which labels perineuronal nets, and parvalbumin, which is expressed in a class of strong inhibitory neurons. By contrast, prefrontal limbic areas were enriched in the enzyme calcium/calmodulin-dependent protein kinase II (CaMKII), known to enhance plasticity. Eulaminate areas have more elaborate laminar architecture than limbic areas and showed the opposite trend: they were enriched in markers of stability and had lower expression of the plasticity-related marker CaMKII. The expression of glial fibrillary acidic protein (GFAP), a marker of activated astrocytes, was also higher in limbic areas, suggesting that cellular stress correlates with the rate of circuit reshaping. Elevated markers of plasticity may endow limbic areas with flexibility necessary for learning and memory within an affective context, but may also render them vulnerable to abnormal structural changes, as seen in neurologic and psychiatric diseases.

Sortilin 1 (Sort1) is a member of the Vps10p domain intracellular trafficking receptor family. Genetic variations of the SORT1 gene are strongly associated with plasma cholesterol levels in humans. Recent studies have linked Sort1 to regulation of cholesterol metabolism in hepatocytes and pro-inflammatory response in macrophages, but the tissue-specific roles of Sort1 in lipid metabolism have not been well defined. We developed Sort1 floxed mice and investigated the development of Western diet (WD)-induced steatosis, hepatic inflammatory response, and hyperlipidemia in hepatocyte Sort1 KO mice and myeloid cell Sort1 KO mice. Our findings suggest that hepatocyte Sort1 deficiency attenuated diet-induced hepatic steatosis and hypercholesterolemia in mice. In contrast, myeloid Sort1 deficiency did not reduce hepatic cytokine expression or plasma cholesterol levels, but exacerbated hepatic triglyceride accumulation in WD-fed mice. Finally, we showed that treating WD-fed mice with an orally bioavailable Sort1 inhibitor, AF38469, decreased plasma cholesterol and hepatic cytokine expression. AF38469 treatment did not affect diet-induced obesity or insulin resistance, but was associated with reduced hepatic VLDL secretion and higher hepatic cholesterol 7á-hydrolase expression in WD-fed mice. In conclusion, findings from this study suggest that Sort1 loss-of-function in hepatocytes contributes to lower plasma cholesterol, and pharmacological inhibition of Sort1 attenuates diet-induced hypercholesterolemia in mice.

Increasing evidence supports the hypothesis that soluble misfolded protein assemblies contribute to the degeneration of postmitotic tissue in amyloid diseases. However, there is a dearth of reliable nonantibody-based probes for selectively detecting oligomeric aggregate structures circulating in plasma or deposited in tissues, making it difficult to scrutinize this hypothesis in patients. Hence, understanding the structure-proteotoxicity relationships driving amyloid diseases remains challenging, hampering the development of early diagnostic and novel treatment strategies. We report peptide-based probes that selectively label misfolded transthyretin (TTR) oligomers circulating in the plasma of TTR hereditary amyloidosis patients exhibiting a predominant neuropathic phenotype. These probes revealed that there are much fewer misfolded TTR oligomers in healthy controls, in asymptomatic carriers of mutations linked to amyloid polyneuropathy, and in patients with TTR-associated cardiomyopathies. The absence of misfolded TTR oligomers in the plasma of cardiomyopathy patients suggests that the tissue tropism observed in the TTR amyloidoses is structure-based. Misfolded oligomers decrease in TTR amyloid polyneuropathy patients treated with disease-modifying therapies (tafamidis or liver transplant-mediated gene therapy). In a subset of TTR amyloid polyneuropathy patients, the probes also detected a circulating TTR fragment that disappeared after tafamidis treatment. Proteomic analysis of the isolated TTR oligomers revealed a specific patient-associated signature composed of proteins that likely associate with the circulating TTR oligomers. Quantification of plasma oligomer concentrations using peptide probes could become an early diagnostic strategy, a response-to-therapy biomarker, and a useful tool for understanding structure-proteotoxicity relationships in the TTR amyloidoses.

In this study was compared the mental health status of 47 multiple sclerosis patients with silver/mercury tooth fillings (amalgams) to that of 50 patients with their fillings removed. On the Beck Depression Inventory the multiple sclerosis subjects with amalgams suffered significantly more depression while their scores on the State-Trait Anger Expression Inventory indicated the former group also exhibited significantly more anger. On the SCL-90 Revised, subjects with amalgam fillings had significantly more symptoms of depression, hostility, psychotism, and were more obsessive-compulsive than the patients with such fillings removed. On a questionnaire containing 18 mental health symptoms multiple sclerosis subjects with amalgam fillings reported a history of 43% more symptoms than those without amalgam fillings over the past 12 months. These data suggested that the poorer mental health status exhibited by multiple sclerosis subjects with dental amalgam fillings may be associated with mercury toxicity from the amalgam.

It is important that hazardous excavated sedimentary and metamorphic rocks are treated appropriately and reused without posing an environmental risk. Up-flow column leaching tests were conducted to examine whether arsenic leaching behavior varied among five hazardous excavated sedimentary and metamorphic rocks (two mudstones, clay sediment of marine origin, slate, and black schist) and to determine whether the potential amount of arsenic leaching could be estimated based on the arsenic-bearing mineral phases in the rock. Changes in arsenic concentration with pore volume (PV) showed the same pattern across all rock types, except for one that contained an extremely low amount of water-soluble arsenic, exhibiting an initial increase to reach a peak, followed by a decrease. The arsenic amounts leached before and after the PV at which the arsenic concentration peaked, corresponded to 88% ± 20% of the amount of arsenic fraction 1 obtained by sequential extraction and 76% ± 10% of the amount of arsenic fraction 2, respectively, while the potential amount of arsenic leaching corresponded to 65-89% of the summed total of arsenic fractions 1 + 2. These findings indicate that arsenic exhibits the same leaching behavior among different types of hazardous excavated sedimentary and metamorphic rocks except where extremely low amounts of water-soluble arsenic are present and that the potential amount of arsenic leaching can be approximated by calculating the summed total of arsenic fractions 1 + 2, which allows us to estimate the minimum amount of material required for treatments such as immobilization conducted to prevent arsenic leaching.

The effect of calcium supplementation on absorption and retention of cadmium in the suckling period was evaluated in Wistar rat pups of both sexes. Animals were maintained in the litters with the mother rats and supplemented with 1%, 3% or 6% calcium (as CaHPO4 x 2H2O) in cow's milk by artificial feeding from day of birth 6 through 14. All rats were exposed to cadmium (as CdCl2 x H2O) either orally or parenterally. Oral cadmium dose of 0.5 mg/kg body weight a day was administered through nine-day period of calcium supplementation and parenteral cadmium dose was injected subcutaneously in a single dose of 0.5 mg Cd/kg body weight prior to calcium supplementation. On experimental day 10 (at the age of pups of 15 days) all animals were killed and the liver, kidneys, brain and carcass (body without organs and skin) were removed for element analyses. Cadmium and essential elements calcium, zinc and iron were analysed in the tissues by atomic absorption spectrometry. Results showed that after oral exposure cadmium concentrations in all calcium-supplemented groups were significantly decreased in the organs and carcass and that the effect was dose-related. No such effect of calcium was found after parenteral cadmium exposure. Calcium supplementation per se significantly increased calcium concentration in the carcass and had no effect on iron in organs and zinc in carcass. It was concluded that calcium supplementation during the suckling period could be an efficient way of reducing oral cadmium absorption and retention without affecting tissue essential trace element concentrations.

BACKGROUND: B7H3 is an immuno-stimulatory glycoprotein that is overexpressed in cancer. However, its functional contributions to cancer development and progression are not well understood. In several reports, it was demonstrated that B7H3 reprograms lipid metabolism and regulates glucose metabolism. Isocitrate dehydrogenase 1 (IDH1), a metabolic enzyme in the TCA cycle, its reaction product is involved in lipid synthesis. Thus, we aimed to identify a novel marker to predict the prognosis of CRC patients and to investigate the relationship between IDH1 and B7H3.METHODS: We analyzed IDH1 and B7H3 expression levels in 225 CRC specimens by immunochemistry. Moreover, in vitro studies were performed to demonstrate the correlation between IDH1 and B7H3.RESULTS: Among 225 tissues, the positive rates of IDH1 and B7H3 were 37.8% (85/225) and 87.6% (197/225), respectively. In CRC samples, IDH1 significantly correlated with B7H3 expression (P = 0.044). Moreover, multivariate analyses revealed that high expression of both B7H3 and IDH1 and a high tumor grade were related to the prognosis of CRC patients. Kaplan-Meier survival analysis revealed that patients with co-expression of IDH1 and B7H3 had a poor overall survival. In SW480B7H3-EGFP cells, which highly express B7H3, IDH1 was up-regulated. Similarly, knockdown of B7H3 expression in Caco-2-shB7-H3 contributed to reduced IDH1 levels.CONCLUSIONS: Although IDH1 and B7H3 cannot be used as independent prognostic factors, co-expression of IDH1 and B7H3 significantly correlated with the prognosis of CRC patients and may serve as a combined predictive marker. Thus, the correlation between IDH1 and B7H3 has been proven in vivo and in vitro.

OBJECTIVES: The high resolution clinic (HRC) is an outpatient care process by which treatment and diagnosis are established, recorded, and completed in a single day. The aim of this study was to assess the extent to which patients with medical conditions may benefit from a single consultation system.MATERIAL AND METHODS: A descriptive study of 795 first visit events, randomly selected as high-resolution consultations in cardiology, gastroenterology, internal medicine, and chest diseases. A discussion is presented on the percentage of patients who benefited from HRC and the complementary tests performed.RESULTS: A total of 559 (70%, 95% CI: 67-73%) of all first visits became HRCs, and 483 (61%, 95% CI: 57%-64%) required a diagnostic test that was reviewed on the same day. There were differences between medical consultations (86% in cardiology versus 44% in gastroenterology consultations, P<.001). Performing a test on the same day significantly increased the percentage of HRCs (49 versus 22%, P<.001). Ischaemic heart disease, dyspepsia, headache, and asthma were the conditions most commonly leading to HRC. The most common tests were cranial tomography, blood analysis, and ultrasound.CONCLUSIONS: Medical consultations may largely benefit from an HRC system, only requiring some organisational changes and no additional costs.

The groundwater of Nalgonda district is well known for its very high fluoride content for the past five decades. Many researchers have contributed their scientific knowledge to unravel causes for fluoride enrichment of groundwater. In the present paper, an attempt has been made to relate the high fluoride content in the groundwater to hydrogeochemical characterization of the water in a fracture hard rock terrain--the Wailpally watershed. Groundwater samples collected from all the major geomorphic units in pre- and post-monsoon seasons were analyzed for its major ion constituents such as Ca(2+), Mg(2+), Na(+), K(+), CO3-, HCO3-, Cl(-), SO4(-2), NO3-, and F(-). The groundwaters in the watershed have the average fluoride content of 2.79 mg/l in pre-monsoon and 2.83 mg/l in post-monsoon. Fluoride concentration in groundwater does not show perceptible change neither with time nor in space. The ionic dominance pattern is in the order of Na(+) > Ca(2+) > Mg(2+) > K(-) among cations and HCO3- Cl(-) > SO4(-2) NO3- F(-) among anions in pre-monsoon. In post-monsoon, Mg replaces Ca(2+) and NO3- takes the place of SO4(-2). The Modified Piper diagram reflect that the water belong to Ca(+2)-Mg(+2)-HCO3- to Na(+)-HCO3- facies. Negative chloralkali indices in both the seasons prove that ion exchange between Na(+) and K(+) in aquatic solution took place with Ca(+2) and Mg(+2) of host rock. The interpretation of plots for different major ions and molar ratios suggest that weathering of silicate rocks and water-rock interaction is responsible for major ion chemistry of groundwater in Wailpally watershed. Chemical characteristics and evolution of this fluoride-contaminated groundwater is akin to normal waters of other hard rock terrain; hence, it can be concluded that aquifer material play an important role in the contribution of fluoride in to the accompanying water. High fluoride content in groundwater can be attributed to the continuous water-rock interaction during the process of percolation with fluoride-bearing country rocks under arid, low precipitation, and high evapotranspiration conditions.

Based on many years of experience in 2009, we developed the original concept of a mixed approach to the treatment of infectious diseases in patients. During 2.5 years(from 2013 to June 2015) to have applied for consultative-diagnostic help of 3965 patients who had not verified the primary diagnosis. The basic principle of verification of the pathology of the removal of various causes immunosuppression. Based on our extensive, research and observation was often found in patients ascaridosis (55%) and giardiasis (65%), as a possible cause of immunosuppression. In 13% of patients was found the mucosal candidiasis. Among frequently and chronically ill persons we identified the active forms of Epstein-Barr virus (quantitative polymerase chain reaction in saliva) in 40%. The criterion for assessing performance immunogram was a decrease of two sigmal deviation from the lower age limit. In the study of neutrophil myeloperoxidase content observed decline (< 60%) in 99 (9.7%) of 1015 patients, indicating a fairly common cause of long-term permit infection in the tissues and persistence C. albicans. In the study of lymphocyte subpopulations often demonstrated reduction in the number of natural killer cells (26.7% of subjects), which shows a decline of one of the most important factors of congenital immunity. Among the humoral immune disorders often noted the decrease of total IgG (2.4%) and its subclass IgG1 (22.1%), indicating a significant diagnostic value determination of IgG subclasses it even with normal serum total. Thus, approximately 76% of patients often suffer set of a decrease immunity. Patients developed with mixed infections caused by various bacterial, fungal, viral and protozoan agents and worms. Immunological study of patients should be redynamics after eliminating the causes immunosuppression and sanitation foci of infection. Only multi-level examination of the patient will determine the final diagnosis and adequate treatment.

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) binds reversibly and with high affinity and specificity to nuclear macromolecules in mouse epidermis. The dissociation constants determined from Scatchard analysis of epidermal nuclei and nuclear macromolecules are 3.58 +/- 0.66 (S.E.) and 2.18 +/- 0.54 nM, respectively. The solubilization of TPA receptors from epidermal nuclei by DNase I was examined. Following a 20-min digestion at 22 degrees, more than a 2-fold increase in specific TPA binding was observed in the supernatant relative to non-nuclease-treated nuclei (0.71 versus 0.32 pmol/mg protein, respectively). Our data indicate that epidermal nuclei contain saturable and specific TPA-binding macromolecules and that these binding components may be associated with regions of chromatin that are preferentially susceptible to nucleolytic cleavage. These data suggest the existence of nuclear receptors for the phorbol ester tumor promoters. These observations may necessitate a more critical assessment of plasma membrane binding as the sole binding site responsible for triggering the multistep process of tumor promotion in mouse epidermis.

Finding causal links between genotype and phenotype is a major issue in biology, even more in mitochondrial biology. First of all, mitochondria form complex networks, undergoing fission and fusion and we do not know how such dynamics influence the distribution of mtDNA variants across the mitochondrial network and how they affect the phenotype. Second, the non-Mendelian inheritance of mitochondrial genes can have sex-specific effects and the mechanism of mitochondrial inheritance is still poorly understood, so it is not clear how selection and/or drift act on mtDNA genetic variation in each generation. Third, we still do not know how mtDNA expression is regulated; there is growing evidence for a convoluted mechanism that includes RNA editing, mRNA stability/turnover, post-transcriptional and post-translational modifications. Fourth, mitochondrial activity differs across species as a result of several interacting processes such as drift, adaptation, genotype-by-environment interactions, mitonuclear coevolution and epistasis. This issue will cover several aspects of mitochondrial biology along the path from genotype to phenotype, and it is subdivided into four sections focusing on mitochondrial genetic variation, on the relationship among mitochondria, germ line and sex, on the role of mitochondria in adaptation and phenotypic plasticity, and on some future perspectives in mitochondrial research. This article is part of the theme issue 'Linking the mitochondrial genotype to phenotype: a complex endeavour'.

BACKGROUND: Achieving normohydration remains a non-trivial issue in haemodialysis therapy. Guiding the haemodialysis patient on the path between fluid overload and dehydration should be the clinical target, although it can be difficult to achieve this target in practice. Objective and clinically applicable methods for the determination of the normohydration status on an individual basis are needed to help in the identification of an appropriate target weight.METHODS: The aim of this prospective trial was to guide the patient population of a complete dialysis centre towards normohydration over the course of approximately 1 year. Fluid status was assessed frequently (at least monthly) in haemodialysis patients (n = 52) with the body composition monitor (BCM), which is based on whole body bioimpedance spectroscopy. The BCM provides the clinician with an objective target for normohydration. The patient population was divided into three groups: the hyperhydrated group (relative fluid overload >15% of extracellular water (ECW); n = 13; Group A), the adverse event group (patients with more than two adverse events in the last 4 weeks; n = 12; Group B) and the remaining patients (n = 27; Group C).RESULTS: In the hyperhydrated group (Group A), fluid overload was reduced by 2.0 L (P < 0.001) without increasing the occurrence of intradialytic adverse events. This resulted in a reduction in systolic blood pressure of 25 mmHg (P = 0.012). Additionally, a 35% reduction in antihypertensive medication (P = 0.031) was achieved. In the adverse event group (Group B), the fluid status was increased by 1.3 L (P = 0.004) resulting in a 73% reduction in intradialytic adverse events (P < 0.001) without significantly increasing the blood pressure.CONCLUSION: The BCM provides an objective assessment of normohydration that is clinically applicable. Guiding the patients towards this target of normohydration leads to better control of hypertension in hyperhydrated patients, less intradialytic adverse events and improved cardiac function.

The authors report an autopsy case with repeated transient ischemic attacks and subsequent strokes demonstrating evidence of artery-to-artery embolism from thrombi generated in the stenotic middle cerebral artery to the distal cortical branches. This case substantiates the existence of intracranial artery-to-artery embolism as a cause of transient ischemic attacks and subsequent strokes, and has potential relevance for therapy.

Mean mental and motor developmental test scores (Bayley scales) at 4, 12, and 24 months of age were determined for 161 graduates of a neonatal intensive care unit during the period 1977 through 1979. Analysis of variance for our data showed significant effects of hyaline membrane disease (HMD) on mental and motor development at 4 months, but disappearance of these effects at 12 and 24 months of age. Conversely, birth weight was not significantly related to developmental performance at 4 months, but was strongly related to both mental and motor performance at 12 and 24 months of age. Gestational age was significantly related to mental performance at 24 months of age. No relationship was found between HMD and major CNS handicapping conditions; both birth weight and gestational age were highly related to the occurrence of neurologic handicaps. Of the three variables assessed, birth weight was the best predictor of neurodevelopmental outcome.

The abilities of isolates of saprophytes (Neurospora crassa, Aspergillus nidulans), an opportunistic human pathogen (Aspergillus fumigatus), an opportunistic insect pathogen (Aspergillus flavus), plant pathogens (Verticillium albo-atrum, Verticillium dahliae, Nectria haematococca), a mushroom pathogen (Verticillium fungicola) and entomopathogens (Verticillium lecanii, Beauveria bassiana, Metarhizium anisopliae) to utilize plant cell walls and insect cuticle components in different nutrient media were compared. The pathogens showed enzymic adaptation to the polymers present in the integuments of their particular hosts. Thus, the plant pathogens produced high levels of enzymes capable of degrading pectic polysaccharides, cellulose and xylan, as well as cutinase substrate, but secreted little or no chitinase and showed no proteolytic activity against elastin and mucin. The entomopathogens and V. fungicola degraded a broad spectrum of proteins (including elastin and mucin) but, except for chitinase, cellulase (V. lecanii and V. fungicola only) and cutinase (B. bassiana only), produced very low levels of polysaccharidases. The saprophytes (Neu. crassa and A. nidulans) and the opportunistic pathogens (A. fumigatus and A. flavus) produced the broadest spectrum of protein and polysaccharide degrading enzymes, indicative of their less specialized nutritional status. V. lecanii and V. albo-atrum were compared in more detail to identity factors that distinguish plant and insect pathogens. V. albo-atrum, but not V. lecanii, grew well on different plant cell wall components. The major class of proteases produced in different media by isolates of V. albo-atrum and V. dahliae were broad spectrum basic (pI > 10) trypsins which degrade Z-AA-AA-Arg-NA substrates (Z, benzoyl; AA, various amino acids; Na, nitroanilide), hide protein azure and insect (Manduca sexta) cuticles. Analogous peptidases were produced by isolates of V. lecanii and V. fungicola but they were specific for Z-Phe-Val-Arg-NA. V. albo-atrum and V. dahliae also produced low levels of neutral (pI ca 7) and basic (pI ca 9.5) subtilisin-like proteases active against a chymotrypsin substrate (Succinyl-Ala2-Pro-Phe-NA) and insect cuticle. In contrast, subtilisins comprised the major protease component secreted by V. lecanii and V. fungicola. Both V. lecanii and V. albo-atrum produced the highest levels of subtilisin and trypsin-like activities during growth on collagen or insect cuticle. Results are discussed in terms of the adaptation of fungi to the requirements of their ecological niches.

We have previously shown that following psoralen photoactivation (PUVA treatment) human dermal fibroblasts undergo long-term growth arrest as well as morphological and functional changes reminiscent of cellular senescence [ 1 ]. In the absence of molecular data on what constitutes normal senescence, it has been difficult to decide whether these PUVA-induced changes reflect cellular senescence or rather a mimic thereof. We herein report that PUVA-induced growth arrest, the senescent phenotype with long-term induction of senescence-associated beta-galactosidase, as well as increased expression of matrix metalloprotease-1 are fully reversible at days 100 to 130 post PUVA treatment in four independently tested fibroblast strains. The late returning growth capacity in PUVA-treated fibroblasts is not due to immortalization, as shown by continued lack of telomerase activity, accelerated telomere shortening, and a decrease in overall growth rates in fibroblasts in their regrowing phase post PUVA treatment. Lack of anchorage-independent growth additionally suggests that the cells are also not tumorigenically transformed. Collectively, our data suggest that PUVA-induced changes do not fully reflect replicative senescence but rather represent a long-term transient phenocopy of senescence. The model reported here is particularly suited to elucidating mechanisms underlying long-term transient growth arrest, the related functional changes, and the release of cells thereof.

Wireless microdevices powered by ultrasound energy have been fabricated to measure and telemeter tissue impedance spectrums for applications in peripheral vascular disease monitoring. The system is characterized by simplicity of the implant consisting of only two electrical components. Ex vivo testing shows the potential for constructing tissue impedance spectrum plots over the range from 10 Hz to 10 kHz by a device less than 1 mm in diameter and 1 cm long. The neurostimulator microdevice was powered by continuous waveform 650 kHz ultrasound with a swept-frequency amplitude modulation. The system was operated at safe ultrasound power levels on the order of 10-100 mW/cm(2). The device proved to be sensitive and able to measure tissue impedances over a broad range. Volume conducted signals carrying impedance information from the microdevice were remotely detected by surface biopotential electrodes.

Vegetative propagation through somatic embryogenesis is critical in conifer biotechnology towards multivarietal forestry that uses elite varieties to cope with environmental and socio-economic issues. An important and still sub-optimal process during in vitro maturation of somatic embryos (SE) is the biosynthesis and deposition of storage proteins, which are rich in amino acids with high nitrogen (N) content, such as arginine. Mobilization of these N-rich proteins is essential for the germination and production of vigorous somatic seedlings. Somatic embryos accumulate lower levels of N reserves than zygotic embryos (ZE) at a similar stage of development. To understand the molecular basis for this difference, the arginine metabolic pathway has been characterized in maritime pine (Pinus pinaster Ait.). The genes involved in arginine metabolism have been identified and GFP-fusion constructs were used to locate the enzymes in different cellular compartments and clarify their metabolic roles during embryogenesis and germination. Analysis of gene expression during somatic embryo maturation revealed high levels of transcripts for genes involved in the biosynthesis and metabolic utilization of arginine. By contrast, enhanced expression levels were only observed during the last stages of maturation and germination of ZE, consistent with the adequate accumulation and mobilization of protein reserves. These results suggest that arginine metabolism is unbalanced in SE (simultaneous biosynthesis and degradation of arginine) and could explain the lower accumulation of storage proteins observed during the late stages of somatic embryogenesis.

At least one-third of patients meeting clinical criteria for Lynch syndrome will have no germline mutation and constitutional epimutations leading to promoter methylation of MLH1 have been identified in a subset of these patients. We report the first case of constitutional MLH1 promoter methylation associated with a colonic polyposis syndrome in a 39 year-old man with a family history of colorectal cancer (CRC) and a personal history of 21 polyps identified over 8 years as well as the development of two synchronous CRCs over 16 months who was evaluated for a hereditary cancer syndrome. Immunohistochemistry (IHC) of multiple tumors showed absent MLH1 and PMS2 expression, though germline testing with Sanger sequencing and multiplex ligation-dependent probe amplification of these mismatch repair genes (MMR) genes was negative. A next generation sequencing panel of 29 genes also failed to identify a pathogenic mutation. Hypermethylation was identified in MLH1 intron 1 in tumor specimens along with buccal cells and peripheral white blood cells, confirming the diagnosis of constitutional MLH1 promoter methylation. This case highlights that constitutional MLH1 methylation should be considered in the differential diagnosis for a polyposis syndrome if IHC staining shows absent MMR gene expression.

Postural effects on water excretion are known to be increased in patients with cervical spinal cord injury and may result in marked impairment of the ability to excrete a water load, especially in erect posture. Both vasopressin-dependent and vasopressin-independent mechanisms have been implicated. To assess the roles of these mechanisms and further identify the factors involved in the renal response to erect posture, sustained water loading studies were performed on 11 quadriplegic subjects and 9 healthy control subjects, supine and erect (sitting). Renal blood flow was assessed by p-aminohippurate clearance (CPAH) measurements in 7 quadriplegic and 5 control subjects. During maximal water diuresis, plasma vasopressin concentrations were reduced to unquantifiable levels in all subjects. Osmolar clearance, free water clearance (CH2O), and distal delivery of filtrate (DDF) were all lower in quadriplegic than in control subjects, supine and erect. The relationship between CH2O and DDF was the same in quadriplegic as in control subjects and was not altered by change in posture in either group. Creatinine clearance and CPAH were lower in erect than in supine posture in quadriplegic subjects but not in control subjects. We conclude that impairment of water excretion in stable normonatremic quadriplegic subjects can be attributed primarily to vasopressin-independent mechanisms involving reduced filtrate delivery to diluting segments of the renal tubules rather than to resistance to normal suppression of vasopressin release.

The serine-threonine kinase Akt1 promotes cell survival through inhibition of apoptosis. One of the potential downstream targets of Akt1 is p70 S6 kinase, p70(S6K), an enzyme implicated in the regulation of protein synthesis. In this study, we investigated the changes in total and phosphorylated levels of Akt1 and p70(S6K) during transient focal ischemia. Male Wistar rats were subjected to 2 h of middle cerebral artery occlusion followed by 1, 4, and 24 h of reperfusion. The expression of total and phosphorylated forms of Akt1 and p70(S6K) were examined by Western blot analysis. Phosphorylation of Akt1 on Ser473 transiently increased at 1 and 4 h of reperfusion, whereas phosphorylation of Akt1 on Thr308 was reduced during reperfusion. The levels of total Akt1 remained unchanged at 1 and 4 h of reperfusion, but decreased significantly at 24 h of reperfusion. Phosphorylation of p70(S6K) on Thr389 decreased at 1, 4, and 24 h of reperfusion, while the levels of total p70(S6K) protein remained unchanged at 1 and 4 h of reperfusion but decreased at 24 h of reperfusion. The results show that cell survival pathways, such as Akt1 and p70(S6K) signaling, are suppressed after transient focal ischemia, which may contribute to the development of neuronal cell death after an ischemic insult.

Long-term follow-up of 508 patients after various surgical procedures has been conducted. Of the 351 patients were operated on using a paramedian incision, 109 had an upper median laparotomy and the rest 48 had an oblique subcostal incision. Post-operative ventral hernias were registered in 45 (8.86%) patients. The upper median laparotomy herniated in 18 (16.51%) patients, paramedian incision - in 17 (4.84%), and the oblique subcostal incision herniated in 10 (20.83%). Hernia was noticed by the patient within 12 months after the initial operation in 46.67%, of them in 85.71% - within first 6 months. 53,33% of patients were diagnosed with postoperative hernia only after thorough examination. Thus, paramedian incision is considered to be the most preferable access. Postoperative hernias develop within the first 6-12 months postoperatively, later hernia registration is a result of poor examination.

Convalescent outpatient parenteral antibiotic therapy with ceftriaxone was evaluated in an uncontrolled study of 101 children with documented serious bacterial infections, including meningitis. Criteria for outpatient therapy were established to assure that risks of complications from the illness were minimal at the time of discharge from the hospital. Daily physician visits and motivated, capable parents were considered essential in outpatient management. Ceftriaxone was given once daily to children with non-central nervous system infections and once or twice daily intravenously to children with meningitis. The mean durations of therapy for children with non-central nervous system infections and with meningitis were 2.4 and 4.6 days, respectively. No child enrolled in this study was readmitted to the hospital for medical or social reasons. Probable complications of treatment included diarrhea in 13% of children with meningitis and in 6% of children with non-central nervous system infections. One child with meningitis developed pseudomembranous colitis. For children who are infected with bacteria that are highly susceptible to ceftriaxone, single daily dose outpatient therapy is a reasonable option for management if a good clinical response to initial treatment is demonstrated and the risks of complications of the disease process are negligible.

Noradrenaline induced concentration-dependent contractions of pulmonary artery segments from control and monocrotaline-treated rats. There was a significant decrease in the maximum response but not sensitivity in artery segments from monocrotaline-treated rats. At a concentration (10(-6) M) that abolished KCl-induced contraction, nifedipine attenuated but did not abolish, noradrenaline-induced contraction in both groups. However, noradrenaline-induced contraction in artery segments from pulmonary hypertensive rats was more susceptible to inhibition by nifedipine. Bumetanide (10(-4) M), a chloride transport inhibitor and niflumic acid, a chloride channel inhibitor, reduced noradrenaline-induced contraction of the pulmonary artery in control and pulmonary hypertensive groups. These compounds were more effective in ring segments from pulmonary hypertensive rats. It was concluded that activation of chloride channels was involved in noradrenaline-induced contraction and that the contribution of chloride channels was enhanced in pulmonary hypertensive rats.

The scope of this study was investigation the affects of various antioxidants on 1% soybean lecithin-based semen extenders for ram semen cryopreservation. Ejaculates, collected via electrically stimulated ejaculation, that have a thick consistency, rapid wave motion (3-5 on a 0-5 scale) and >75% initial motility were pooled. The pooled samples were split into four equal aliquots as 5 mM Methionine, 5 mM Cysteamine, 1 mM Cysteine and a sample of antioxidant-free control group. Each sample group was diluted to a ratio of 1/5 (semen/extender, v/v) as final concentration and two step dilution method was used for cryopreservation. Extender groups were assessed for sperm motility, plasma membrane functional integrity using hypoosmotic swelling test (HOST), damaged acrosome using FITC-Pisum sativum agglutinin (PSA-FITC) and DNA integrity using terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL). Semen samples also incubated for 6 h in humidified air with 5% CO2 at 39 °C to evaluate post-thaw incubation resilience of semen characteristics. The results showed that freezing and thawing procedures had negative effects on motility (P < 0.05), plasma membrane integrity (P < 0.05) and acrosomal integrity (P < 0.05). After 6 h of incubation time, the Cysteine supplemented extender group yielded significantly higher results than other extender groups in terms of spermatological parameters. Furthermore MDA levels in the antioxidant groups were lower than control group (P < 0.05). Nevertheless, there were no significant differences among antioxidant groups.

Goose down and duck down have very similar appearance but the quality of goose down is better than that of duck down in general. There is a highest allowable limit as specified by the various national standards of feather and down for the percentage of duck feather or down mixed in goose feather or down. Traditional detection method, manual inspection with a high-scale microscope, is labor intensive and not suitable for large-volume samples analysis and on-site rapid testing. In the present paper, visible/near-infrared (NIR) spectroscopy combined with successive projection algorithm (SPA) for characteristic wavelengths selection was used to determinate the content of duck down mixed in goose down. In the range of 450-930 nm, the multiple linear regression (MLR) model established with the 8 characteristic wavelengths selected by SPA achieved good prediction, the correlation coefficient of 0.983, root mean square error of calibration (RMSEC) of 5.44%, and root mean square error of prediction (RMSEP) of 5.75%. Therefore, it is expected to be used for rapid detection of feather and down quality in future.

OBJECTIVES: To determine the spontaneous variability of ventricular arrhythmias (VA) and evaluate anti-arrhythmic efficacy of mexiletine, sotalol, and a mexiletine-sotalol combination in German shepherd dogs (GSD) with inherited arrhythmias.ANIMALS, MATERIALS AND METHODS: 12 affected GSD, median age 20 weeks, received mexiletine (8 mg/kg PO q8 h), sotalol (2.5 mg/kg PO q12 h), and combination therapy for 6 days in random order. Pre- and post-treatment 24 h Holter recordings were acquired, allowing determination of VA variability and reduction in 24 h VA for each treatment. Drug concentrations during each arm were measured.RESULTS: An anti-arrhythmic effect could be inferred if ventricular premature complexes (VPC), ventricular couplets (V(cpl)), ventricular tachycardia runs (VT(runs)) and total ventricular ectopy (VE(tot)) frequency were reduced by 61%, 97%, 98%, and 63% (1 control Holter model), by 53%, 94%, 95%, and 54% (4 control Holter model) and by 54%, 95%, 96% and 56% (3 control Holter model). Combination therapy reduced VPC and VE(tot) in more dogs (5/12 and 6/12) than mexiletine (1/11 and 2/11) or sotalol (2/9 and 1/9) (p < 0.05). The combination therapy reduced the mean number of VPC, V(cpl), and VE(tot). Sotalol monotherapy produced an increase in VT(runs). Plasma mexiletine concentration was higher during combination therapy than with monotherapy.CONCLUSIONS: Combination therapy reduced VPC in affected GSD. Sotalol monotherapy increased VT(runs). Combination therapy increased plasma mexiletine concentrations.

The ability of plant lectins to modify the interactions of the insulin receptor (IR) and insulin-like growth factor (IGF) receptors (IGFRs) with their ligands was investigated. The lectins profoundly affected the competition-binding curves for (125)I-labelled IGF-I and insulin, causing an increase in the affinity of placental IGF1R and IR towards their ligands. This increment was of such a magnitude that it could affect the receptors' specificity towards these ligands. The lower the ligand concentration, the greater was the lectin-induced affinity shift, which suggests potential physiological significance of the effect. The affinity modulation occurred in a lectin-specific and dose-dependent manner. In contrast to IGF1R and IR, the binding of (125)I-labelled IGF-II to its receptors resisted lectin modulation. Here we provide evidence of the possibility of external modulation of the affinity of placental IGF1R and IR via interactions of the receptors' carbohydrate moieties with lectins. The existence of modulators that would selectively inhibit or enhance the binding of IGFs or insulin to their corresponding receptors may have important implications for placental cell responses to these molecules.

UNLABELLED: In Mecklenburg-Vorpommern the Provincial Government has decreed that the laboratory diagnostic examination of applicants for asylum is limited to Salmonella and Shigella. A comprehensive bacteriological and parasitological investigation of 517 applicants for asylum showed a wide pattern of intestinal pathogens. 123 person (23.79%) were infected with pathogenic or facultative pathogenic agents. Regional differences of the extent of distribution and the risks of infestation with pathogens by importation are discussed.CONCLUSION: Limitation of obligatory microbiologic examination to Salmonella and Shigella only is insufficient.