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Twenty derivatives of aphidicolin were tested against HSV (herpes simplex virus), HCMV (human cytomegalovirus) and adenovirus in vitro. In addition, the antiviral activity of aphidicolin (CAS 38966-21-1) in combination with aciclovir (CAS 59277-89-3) or cidofovir (CAS 113852-37-2) against HSV was determined. The antiviral effects were evaluated using plaque reduction assay in Vero cells or human Foreskin Fibroblasts (HFF) for HSV and HCMV, respectively. Combination indexes were calculated using the method of Chou and Talalay. Two derivatives (K14254 and K14266) that are considered to be prodrugs of aphidicolin were shown to inhibit HCMV and HSV replication comparably to aphidicolin. None of the tested substances inhibited adenovirus replication. Aphidicolin acted synergistically with aciclovir in a 1:1 molar ratio and with cidofovir in different ratios. Aphidicolin and its two antiviral active derivatives might represent useful additional tools for antiviral therapy of HSV and HCMV infections, especially in combination with clinically used drugs.

Accidental deaths in childhood were studied between 1978 and 1982 in Metropolitan Dade County. Cases were collected from both traffic and nontraffic accidental fatalities in which the victim was less than 12 years of age. Cases were then subdivided into traffic and nontraffic groups. Each group was then analyzed as to age, race, sex, cause of death, and scene circumstances. A total of 294 cases was studied.

A mouse strain was identified with a recessive genetic lesion, which spontaneously developed a lymphoproliferative autoimmune syndrome exhibiting features of systemic lupus erythematosus. Positional mapping of the disease-associated locus revealed a lesion in Rasgrp1 that prevented the translation of the RasGRP1 protein. T cells from these mice failed to activate Ras or proliferate vigorously following antigen encounter and showed defects in positive selection. Peripheral RasGRP1lag T cells spontaneously adopted a memory phenotype and were able to transfer disease to lymphopenic recipient mice. CD4+ T cells accumulated in the lymphoid tissues of older RasGRP1lag mice and were resistant to activation-induced cell death. RasGRP1lag B cells were functionally normal, but activated B cells were detected in older mice, as were autoantibodies directed against self-antigens. Our findings indicate that Ras signaling pathways are required to maintain T cell tolerance and to prevent autoimmune disease.

Infection of susceptible strains of mice with Friend leukemia virus (FLV) results in a profound depression of cell-mediated immunity as assessed by lymphocyte-mediated cytotoxicity. This depression occurs early in the disease, before the onset of splenomegaly, and is associated with a decline in the susceptibility of splenocytes from FLV-infected mice to lysis by anti-Thy-1. 2 serum and complement. Treatment of splenocytes from FLV-infected mice with neuraminidase restores, in large part, their susceptibility to anti-Thy-1.2 serum as well as their cytolytic capacity. These studies suggest that one early immunosuppressive consequence of infection with FLV involves alteration of the effector T-lymphocyte cell surface.

To date, two structurally related RNA-editing enzymes with adenosine deaminase activity have been identified in mammalian tissue: ADAR1 and ADAR2 [Bass B. I. et al. (1997) RNA 3, 947-949]. In rodents, ADAR2 undergoes alternative RNA splicing, giving rise to two splice variants that differ by the presence or absence of a 10-amino-acid insert in the carboxy-terminal catalytic domain. However, the physiological significance of the splicing and its regional and developmental regulation are as yet unknown. The present study examined spatial and temporal patterns of ADAR2 gene transcripts within specific neuronal populations of rat brain. The two rodent ADAR2 isoforms were expressed at comparable levels at all ages examined. rADAR2 messenger RNA expression was first detectable in the thalamic nuclei formation at embryonic day E19. The rADAR2b insert and rADAR2a splice probes produced images similar to that of the rADAR2 pan probe. At birth, rADAR2a messenger RNA splice variants were abundantly expressed in the thalamic nuclei. No signal for any probe was detectable in other brain regions, including neocortex, hippocampus, striatum and cerebellum at this stage of development. During the first week of postnatal life, rADAR2 messenger RNA expression (detected with the pan probe) increased gradually in several brain regions, with low expression detected at postnatal day P7 in the olfactory bulb, inferior colliculus, and within the pyramidal and granule cell layers of the hippocampus. Hybridization patterns of the rADAR2a variant probe reached peak expression at about the second week of life, while peak expression of the rADAR2b probe was reached at about the third week of life. At the end of the first week of life (P7), expression of both splice variants was strongest in the thalamic nuclei. By P14, rADAR2 messenger RNA expression was more consolidated in the deeper structures, including the thalamic nuclei and the granule cell layer of the cerebellum. By P21, maximal levels of rADARb expression were observed in the thalamic nuclei, inferior colliculus, cerebellum and pontine nuclei. In the adult, rADAR2 messenger RNA expression was of highest intensity in the thalamic nuclei, with high levels of expression in the olfactory bulb, inferior colliculus, cerebellum and pontine nuclei. At the level of the hippocampus, positive labelling was restricted to the CA3 region of the Ammon's horn and the dentate gyrus, with weak signals in the CA1 subfield. rADAR2 pan expression was at near background levels throughout the neocortex and caudate putamen. In summary, our study shows that ADAR2 messenger RNA expression is regulated in a cell-specific manner throughout development. At early ages, ADAR2 messenger RNA is expressed only within (and restricted to) the thalamic nuclei. By the third postnatal week, expression of the editase enzyme is more widely distributed throughout the olfactory bulb, CA3 and dentate gyrus of the hippocampus, thalamus, inferior colliculus and the molecular cell layer of the cerebellum. ADAR2 is thought to act at specific nucleotide positions in primary transcripts encoding glutamate receptor subunits, thereby altering gating and ionic permeability properties of AMPA- and kainate-activated channels. ADAR2 also acts at pre-messenger RNA encoding the serotonin 5HT-2C receptor to alter G-protein coupling. Thus, RNA editing may be an important mechanism for fine-tuning of the physiological and pharmacological properties of transmitter receptors of the central nervous system.

This study investigates variation in body growth (cross-sectional height and weight velocity) among a sample of 22 small-scale societies. Considerable variation in growth exists among hunter-gatherers that overlaps heavily with growth trajectories present in groups focusing more on horticulture. Intergroup variation tends to track environmental conditions, with societies under more favorable conditions displaying faster growth and earlier puberty. In addition, faster/earlier development in females is correlated with higher mortality. For example, African "Pygmies," Philippine "Negritos," and the Hiwi of Venezuela are characterized by relatively fast child-juvenile growth for their adult body size (used as a proxy for energetic availability). In these societies, subadult survival is low, and puberty, menarche, and first reproduction are relatively early (given their adult body size), suggesting selective pressure for accelerated development in the face of higher mortality. In sum, the origin and maintenance of different human ontogenies may require explanations invoking both environmental constraints and selective pressures.

Production of platelet-activating factor by washed rabbit platelets under stimulation with the ionophore A23187 was investigated utilizing two groups of platelet preparations. The first platelet preparation contained 0.03 +/- 0.02% contaminating white cells, while the second preparation contained 0.48 +/- 0.27% white cells. The latter preparation produced platelet-activating factor, mainly 1-hexadecyl-2-acetyl-sn-glycero-3-phosphocholine, 8.3 +/- 6.3 pmol (mean +/- standard deviation) with a range of 2.6 to 21.4 pmol (n = 9), followed by small quantities of 1-octadecenyl- and 1-octadecyl-2-acetyl-sn-glycero-3-phosphocholine. In contrast, there was no production of 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine by the former platelet preparation having 0.03% leukocytes. These quantitative analyses were carried out by the selected ion monitoring technique and it was concluded that it is necessary to consider the presence of contaminating white cells in studies on the production of platelet-activating factor by platelets.

Previous studies have detected gamma delta T cells in multiple sclerosis and experimental allergic encephalomyelitis (EAE) lesions but their role remains obscure. In the present study, we assessed gamma delta T cell dynamics and distribution in spleen and central nervous system (CNS) from mice with relapsing-remitting EAE, and studied the effect of depleting these cells on clinical and pathologic expression of disease using the mAb GL3. By immunohistochemistry and FACS analysis, striking disease-related changes were observed in the gamma delta T cell population in the CNS. FACS analysis showed that while gamma delta T cells remained low in the spleen (approximately 2% total CD3+ T cells) at all stages, in the CNS they increased to approximately 12% at the height of the acute attack, fell to approximately 5% during the recovery phase, but rose again to approximately 12% during the chronic phase. In animals in which gamma delta T cells were depleted immediately before the onset of acute disease, or during the chronic stage, a striking and significant reduction in the severity of the clinical signs was observed that was associated with a decrease in the percentage of CD3+/gamma delta T cells in the CNS. In depleted animals a statistically significant reduction in inflammation and demyelination was noted during the acute stage, but only marginal effects on these disease parameters were found in the chronic phase. Taken together, the data support the conclusion that gamma delta T cells play an important role in the pathogenesis of EAE in mice during both acute and chronic/progressive phases of the disease process.

BACKGROUND: Muscle relaxants (MR) are responsible for 59% of peroperative anaphylactic reactions. A major issue would be to determine whether a systematic preoperative screening in the general population should be recommended.OBJECTIVE: The purpose of the study was to evaluate the prevalence of muscle relaxant sensitivity in a sample of the general population and to assess the role of possible risk factors.METHODS: Two hundred and fifty-eight subjects, aged 20-40 years, visiting a health care centre for a check-up were evaluated. Protocol included a questionnaire (occupation, symptoms of atopy, previous surgery, history of drug allergy), skin-prick tests to four commercial muscle relaxants and measurement of specific IgE against quaternary ammonium ions. Atopy was evaluated by skin-prick tests to common inhalant allergens and by a Phadiatop test.RESULTS: Of the study group, 9.3% had either a positive skin test to one or more muscle relaxant or a presence of specific IgE to quaternary ammonium ions. No risk factor was identified in the studied group.CONCLUSION: Since the rate of MR sensitivity is much higher than the anticipated rate of peroperative reactions due to allergy, a systematic preoperative screening for MR allergy should not be recommended for adults in a general population.

OBJECTIVES: To evaluate the performance of first-trimester screening for trisomy 21 by a combination of maternal age, fetal nuchal translucency (NT) and maternal serum free beta-human chorionic gonadotropin (beta-hCG) and pregnancy-associated plasma protein-A (PAPP-A). In addition, the potential impact of a new individual risk-orientated two-stage approach to first-trimester screening was examined.METHODS: First-trimester combined screening for trisomy 21 was carried out in 75 821 singleton pregnancies with live fetuses at 11 + 0 to 13 + 6 gestational weeks. The detection and false-positive rates for different risk cut-offs were calculated. To examine the potential impact of an individual risk-orientated two-stage approach to first-trimester screening it was assumed that, after first-trimester combined screening, chorionic villus sampling (CVS) would be performed in all patients with a risk estimate of 1 in 100 or more and in none of those with a risk estimate of less than 1 in 1000. Those in the intermediate-risk category, with a risk estimate of between 1 in 101 and 1 in 1000, would have further assessment of risk by first-trimester ultrasound examination to determine presence/absence of the nasal bone, presence/absence of tricuspid regurgitation or normal/abnormal Doppler velocity waveform in the ductus venosus, and CVS would be performed if their adjusted risk became 1 in 100 or more.RESULTS: Fetal NT and maternal serum free beta-hCG and PAPP-A were successfully measured in all cases. The median maternal age was 31 (range, 13-49) years, the median gestation at screening was 12 (range, 11 + 0 to 13 + 6) weeks and the median fetal crown-rump length was 62 (range, 45-84) mm. Chromosomal abnormalities were identified in 544 pregnancies, including 325 cases of trisomy 21. The estimated risk for trisomy 21 was 1 in 300 or greater in 5.2% of normal pregnancies, in 92.6% of those with trisomy 21, in 88.5% of those with trisomy 18 or 13 and in 85.6% of those with other chromosomal defects. The detection rates for trisomy 21 were about 75% and 80% for respective false-positive rates of 1% and 2%. In the proposed individual risk-orientated two-stage screening for a risk cut-off of 1 in 100 the total false-positive rate would vary with the method used for the second stage of screening from 2.1% for absence of the nasal bone to 2.7% for increased impedance in the ductus venosus and 2.7% for tricuspid regurgitation and the respective detection rates would be 92.0%, 94.2% and 91.7%.CONCLUSIONS: First-trimester combined screening for trisomy 21 is associated with a detection rate of about 90% for a false-positive rate of 5%. Individual risk-orientated two-stage screening for trisomy 21 can potentially identify, in the first trimester of pregnancy, more than 90% of affected fetuses for a false-positive rate of 2-3%.

AIM: To evaluate clarithromycin 500 mg t.d.s., tripotassium dicitrato bismuthate 240 mg b.d. and omeprazole 20 mg b.d. for 7 days as a Helicobacter pylori treatment regimen.METHODS: The H. pylori status of dyspeptic patients undergoing endoscopy was assessed by histology, culture and rapid urease testing of biopsies and by 13C-urea breath test. Fifty patients who were H. pylori-positive were treated with the above treatment regimen for 7 days. Those patients with active duodenal ulcers present at endoscopy were given omeprazole 20 mg nocte for a further 21 days. Not less than 28 days after completing treatment, all tests were repeated to reassess H. pylori status. Bacterial sensitivity of H. pylori cultures was determined and patients recorded any side-effects.RESULTS: On an intention-to-treat basis, H. pylori infection was cured in 90% (95% CI: 78-96%) of patients. Taste disturbance was experienced by 35% patients. Compliance was excellent, with 96% patients taking more than 95% of tablets. Metronidazole resistance was 41% but all cultures were sensitive to clarithromycin.CONCLUSIONS: This 7-day treatment achieved a high level of cure of H. pylori infection with relatively minor side-effects. It may have a role to play, particularly where there is a high level of metronidazole resistance.

The key to developing a successful CVVH course is the involvement of the clinical experts--the staff nurses. It is an ongoing process that requires continual improvement. The CVVH course will help to assure the competency of the ICU nurse in caring for the critically ill patient on CVVH.

BACKGROUND: Visible light (VL) has multiple effects on the skin that currently available sunscreens do not protect against. Polypodium leucotomos extract (PLE) has properties that may offer protection against VL. OBJECTIVES: To determine the effectiveness of PLE in preventing VL-induced effects. METHODS: Twenty-two subjects with Fitzpatrick skin phototype IV-VI were enrolled. On day 0, subjects were irradiated with VL. Clinical Investigator’s Global Assessment (IGA) scoring and spectroscopic evaluations were performed immediately, 24 hours, and 7 days after irradiation. Subjects then received a 28-day supply of PLE (480 mg daily). Irradiation and evaluation were repeated. Three 4-mm punch biopsies were obtained for immunohistochemistry analysis: one from normal unirradiated skin and the other two twenty-four hours after irradiation, pre- and post-PLE, from sites irradiated with highest dose of VL. RESULTS: All subjects had immediate pigment darkening, persistent pigment darkening, and delayed tanning both pre- and post-PLE. For the highest VL dose (480 J/cm²) spectroscopic assessments demonstrated a statistically significant decrease in persistent pigment darkening and delayed tanning post-PLE. In addition, there was a significant decrease in cyclooxygenase-2, and a trend towards decreases in the markers for cellular damage post-PLE. While there was a trend towards lower IGA scores post-PLE, statistical significance was not reached possibly due to lack of sensitivity of the visual IGA scoring system in detecting small changes. CONCLUSIONS: Spectroscopic data and immunohistochemistry indicate an effect of PLE on visible light induced effects. As such, PLE may be used as an adjuvant to traditional means of photoprotection to protect against the effects of VL. Clinical trial registration number: NCT02904798. J Drugs Dermatol. 2019;18(12):1198-1203.

BACKGROUND: The role of dietary protein in short term satiety is of interest with respect to body weight regulation.AIM: To compare the effects of a high versus a normal soyprotein breakfast on satiety and subsequent energy intake (EI), including 'satiety' hormones and plasma amino acid responses.METHODS: Twenty-five healthy subjects (mean +/- SEM, BMI: 23.9 +/- 0.3 kg/m(2); age: 22 +/- 1 years) received a subject-specific standardized breakfast: a custard with soy as single protein type with either 10/55/35 (normal-protein) or 25/55/20 (high-protein) En% protein/carbohydrate/fat in a randomized, single-blind design. Appetite profile (Visual Analogue Scale, VAS), plasma glucose, insulin, Glucagon-like Peptide 1, ghrelin, and amino acid concentrations were determined for 4 h, determining the sensitive time point to assess EI. Since at 180 min glucose and insulin concentrations still were significantly different, in a second set of experiments subjects received an ad lib lunch at 180 min after the breakfasts; EI was assessed.RESULTS: Overall the 25 En% soy-custard was rated as being more satiating than the 10 En% soy-custard (P < 0.01) and there was a difference at 20 min after breakfast (64 +/- 5 vs. 52 +/- 5 mmVAS, P < 0.05), related to higher postprandial taurine concentrations (P < 0.05). Insulin response was increased more after the 25 En% than after the 10 En% soy-custard (AUC: 7,520 +/- 929 vs. 4,936 +/- 468 mU/l h, P < 0.001). There was no difference in EI (25 En%: 3,212 +/- 280 kJ vs. 10 En%: 3,098 +/- 286 kJ, ns).CONCLUSION: A high soyprotein breakfast is more satiating than a normal soyprotein breakfast related to elevated taurine and insulin concentrations.

Selected characteristics of disfluent conversational utterances with and without disfluency clusters were examined in 14 children who stutter (CWS) and 14 children who do not stutter (CWNS). For CWS, utterances with disfluency clusters contained significantly more syllables and clausal constituents than disfluent utterances without clusters, which, in turn, contained significantly more syllables, clauses, and clausal constituents than fluent utterances. For both groups of children, disfluency clusters coincided significantly more often with utterance or clause onset than they did with grammatical constituents located elsewhere within an utterance. CWNS produced a significantly greater percentage of disfluency clusters that contained grammatical revision than did CWS. No significant between-group differences were observed in terms of the number of syllables, clauses, or clausal constituents within cluster-inclusive utterances. Findings are taken to suggest that disfluency clusters are typically produced within the most complex linguistic contexts and that they reflect the effects of producing multiple syntactic constituents within an utterance.

Orbitofrontal Cortex (OFC) structural abnormality in schizophrenia has not been well characterized, probably due to marked anatomical variability and lack of consistent definitions. We previously reported OFC sulcogyral pattern alteration and its associations with social disturbance in schizophrenia, but OFC volume associations with psychopathology and cognition have not been investigated. We compared chronically treated schizophrenia patients with healthy control (HC) subjects, using a novel, reliable parcellation of OFC subregions and their association with cognition, especially the Iowa Gambling Task (IGT), and with schizophrenic psychopathology including thought disorder. Twenty-four patients with schizophrenia and 25 age-matched HC subjects underwent MRI. OFC Regions of Interest (ROI) were manually delineated according to anatomical boundaries: Gyrus Rectus (GR); Middle Orbital Gyrus (MiOG); and Lateral Orbital Gyrus (LOG). The OFC sulcogyral pattern was also classified. Additionally, MiOG probability maps were created and compared between groups in a voxel-wise manner. Both groups underwent cognitive evaluations using the IGT, Wisconsin Card Sorting Test, and Trail Making Test (TMT). An 11% bilaterally smaller MiOG volume was observed in schizophrenia, compared with HC (F(1,47) = 17.4, P = 0.0001). GR and LOG did not differ, although GR showed a rightward asymmetry in both groups (F(1,47) = 19.2, P < 0.0001). The smaller MiOG volume was independent of the OFC sulcogyral pattern, which differed in schizophrenia and HC (chi2 = 12.49, P = 0.002). A comparison of MiOG probability maps suggested that the anterior heteromodal region was more affected in the schizophrenia group than the posterior paralimbic region. In the schizophrenia group, a smaller left MiOG was strongly associated with worse 'positive formal thought disorder' (r = -0.638, P = 0.001), and a smaller right MiOG with a longer duration of the illness (r = -0.618, P = 0.002). While schizophrenics showed poorer performance than HC in the IGT, performance was not correlated with OFC volume. However, within the HC group, the larger the right hemisphere MiOG volume, the better the performance in the IGT (r = 0.541, P = 0.005), and the larger the left hemisphere volume, the faster the switching attention performance for the TMT, Trails B (r = -0.608, P = 0.003). The present study, applying a new anatomical parcellation method, demonstrated a subregion-specific OFC grey matter volume deficit in patients with schizophrenia, which was independent of OFC sulcogyral pattern. This volume deficit was associated with a longer duration of illness and greater formal thought disorder. In HC the finding of a quantitative association between OFC volume and IGT performance constitutes, to our knowledge, the first report of this association.

The City of Milwaukee Health Department piloted a short-term, near real-time syndromic surveillance and communication tool by using an existing secure regional Internet infrastructure. Voluntary, active syndromic case reporting by hospital Emergency Departments was combined with other data streams, including clinical laboratory reports of communicable disease, hospital emergency room diversions, ambulance runs, medical examiner reports of unusual or suspicious deaths, poison control and nursing hotline call volumes, and pharmacy over-the-counter sales. These data were aggregated into a "Surveillance Dashboard" format that was used to communicate community syndromic health trends to hospitals, Emergency Departments, and other providers using a secure Internet technology. Emergency Departments at 8 area hospitals reported a total of 314 cases meeting syndromic criteria from 26,888 patient encounters. Participants were satisfied with data entry and communications. All participating Emergency Departments received e-mail and text pager alerts sent by the Milwaukee Health Department. No unexplained findings or suggestions of an early outbreak were reported through syndrome surveillance for the 4-week duration of the project. Similar surveillance and communications systems could provide multiple benefits to Emergency Department workflow and management, as well as to public health and emergency response.

INTRODUCTION: The Nance appliance is widely considered to be an efficient method of anchorage reinforcement; however, much of the perceived advantage is based on clinical judgment. The aim of this study was to assess the amounts of anchorage loss and desired tooth movement associated with the Nance appliance.METHODS: The mandibular arches of 7 beagle dogs were used. The first and third premolars were extracted. Reference miniscrews were placed at the first premolar sites as stable references to measure the amounts of anchorage loss and desired tooth movement. Four beagles were fitted with custom-made Nance appliances on the fourth premolars and orthodontic bands on the second premolars (Nance group). Three beagles were fitted with orthodontic bands on the second and fourth premolars with no anchorage reinforcement (control group). The second premolars were retracted over 15 weeks in both groups. The amounts of second premolar movement (desired tooth movement) and fourth premolar movement (anchorage loss) were recorded at 5, 10, and 15 weeks. The percentages of desired tooth movement and anchorage loss to the total space closure were calculated.RESULTS: The mean desired tooth movement was significantly more in the Nance group than in the control group at 10 weeks (P <0.05) but was not significantly different at 5 and 15 weeks. The mean percentages of anchorage loss to the total space closure at 15 weeks were 45.7% in the control group and 28.8% in the Nance group. The Nance group had 16.9% less anchorage loss and 16.6% more desired tooth movement than did the control group at 15 weeks (P <0.05). Most of the anchorage loss (80%) in the Nance group occurred during the first 10 weeks.CONCLUSIONS: The Nance appliance did not provide absolute anchorage, but there was significantly less anchorage loss with it than in the control group. The majority of anchorage loss occurred during the first 10 weeks in the Nance group.

Background & Objectives: This pilot study aimed to evaluate the efficacy of memory training and health training intervention over a 24-month period in people with probable mild cognitive impairment (MCI). Research Design & Methods: Based on the accepted criteria, and the neuropsychiatric measures used in the trial, MCI was defined as a subjective change in cognition, impairment in episodic memory, preservation of independence of functional abilities, and no dementia. Without a neurological assessment, laboratory tests, and psychometric evaluation combined, some of our participants may have had dementia that we were unable to detect through neuropsychological testing. Of the 263 total participants, 39 met criteria for a diagnosis of MCI. There were 19 adults in the memory and 20 in health training conditions. Both groups received twenty hours of classroom content that included eight hours of booster sessions at three months post intervention. Hierarchical linear models (HLM) and standardized regression-based (SBR) analyses were used to test the efficacy of the intervention on immediate recall, delayed recall, subjective memory complaints, and memory self-efficacy. Age, education, depression, racial group, ethnic group, MMSE score, and baseline performance were included as covariates. Results: Over 24 months, the MCI group in the memory training condition showed better objective and subjective memory outcomes compared with the MCI group in the health training condition. Conclusions: Senior WISE Memory training delivered to individuals with MCI was able to forestall the participants' declining cognitive ability and sustain the benefit over two years in both subjective and objective memory function.

AIMS: To determine outcomes utilizing thymoglobulin and sirolimus immunosuppression, with early steroid withdrawal in low-immune responder pancreas-kidney (SPK) recipients, and conversion from cyclosporine (CsA) to mycophenolic acid (MPA) in all recipients at 6 months posttransplantation.METHODS: SPK recipients received thymoglobulin, sirolimus, and reduced-dose CsA immunosuppression. Low immune responders (non-African-Americans with a pretransplant PRA < 30%) were withdrawn from prednisone on posttransplant day 5 and high immune responders were continued on prednisone. All recipients were converted from CsA to MPA at 6 months posttransplantation. During conversion, recipient immune response was monitored by flow PRA and a T-cell stimulation assay (Cylex).RESULTS: With a mean follow-up of 9 +/- 4 months, one pancreas was lost to pancreatitis, with no patient or kidney losses and no acute rejection episodes. All eight low immune responder patients were steroid-free at 9 +/- 5 months posttransplantation. Seven patients (five low and two high immune responders) with at least 6-month follow-up were converted from CsA to MPA. One high immune responder with a pretransplant PRA of 43% remained with a PRA of 53% +/- 2% postconversion. The second high immune responder had a pretransplant PRA of 34% and a postconversion PRA of 0%. The five low immune responders had a mean pretransplant PRA of 16% +/- 15% and a postconversion PRA of 0% (P < .01). The Cylex assay resulted in 67% low responsiveness for both high and low immune responders.CONCLUSION: Thymoglobulin induction with sirolimus maintenance therapy permitted immunosuppression minimization in selected pancreas transplant recipients. Posttransplant evaluation revealed a diminished (regulated) immune response in six of seven patients.

Eleven patients with essential hypertension were given hydralazine (25 mg b.i.d.) for 2 weeks, hydralazine and propranolol (80 mg b.i.d.) for the next 2 weeks, and propranolol alone for the last 2 weeks. The changes in the renal and systemic circulation elicited by the drugs were studied before start of medication and bi-weekly during the treatment, using non-invasive radioisotope techniques. Hydralazine alone did not alter mean arterial blood pressure (MAP), heart rate (HR), cardiac index (CI), effective renal plasma flow (ERPF), peripheral renin activity (PRA) and plasma aldosterone (Aldo) but when propranolol was added MAP fell 15.2%, HR 22.5% and CI 18.4%, while ERPF was unchanged. When hydralazine was withdrawn and propranolol was given alone, ERPF decreased 13.2%. Plasma aldosterone was unchanged, whereas PRA decreased during propranolol treatment. The reduction in ERPF elicited by propranolol, was highly significant (P less than 0.01). From the test sequence it appears that dihydralazine prevents this effect of propranolol on kidney function. These findings might have a direct bearing on the choice of antihypertensive treatment.

The Arp2/3 complex mediates formation of complex cellular structures such as lamellipodia by nucleating branched actin filaments. Arp2/3-complex activity is precisely controlled by over a dozen regulators, yet the structural mechanism by which regulators interact with the complex is unknown. GMF is a recently discovered regulator of the Arp2/3 complex that can inhibit nucleation and disassemble branches. We solved the structure of the 240-kDa assembly of Mus musculus GMF and Bos taurus Arp2/3 complex and found that GMF binds the barbed end of Arp2, overlapping with the proposed binding site of WASP-family proteins. The structure suggests that GMF can bind branch junctions in the manner that cofilin binds filament sides, consistent with a modified cofilin-like mechanism for debranching by GMF. The GMF-Arp2 interface reveals how the ADF-H actin-binding domain in GMF is exploited to specifically recognize Arp2/3 complex and not actin.

5-HT receptor positively coupled to adenylyl cyclase in striatal neurones in culture does not correspond to the 5-HT4 receptor. 5-HT induces an increase in cAMP level with an EC50 of 125 nM. 5-HT agonists displayed the following rank order of potencies 5-HT > LSD > 5-MeOT > 5-CT. 8-OH-DPAT, RU 24969 and cisapride were inactive. The most efficacious antagonists were methiothepin and tricyclic antipsychotic drugs (clozapine, amitriptyline and nortryptyline). The pharmacological profile defined by both functional studies (cAMP level) and binding experiments ([125I]-LSD binding), and its localization in striatal neurones are in favour of the presence of the recently cloned 5-HT6 receptor in these cells.

Dopamine receptor-coupled stimulation of inositol phosphate formation has been characterized extensively, but little is known about the diacylglycerol arm of this dual-signaling pathway. This study examined several parameters of cytidine diphosphate-diacylglycerol (CDP-DG) accumulation as an index of agonist-stimulated DG formation. Rat brain slices pre-labeled with 5-[3H]cytidine were incubated with various test agents in the presence of LiCl and accumulated CDP-DG analyzed. Dopamine and SKF38393 significantly and dose-dependently stimulated CDP-DG accumulation. SKF38393 responses were inhibited by neomycin and reversed by myo-inositol or by exclusion of LiCl. Compared to inositol phosphate formation in 2-[3H]inositol-prelabeled slices, the CDP-DG responses were proportionately greater, while the agonist EC50 values were similar between the two assays. The D1-receptor antagonist SCH23390 inhibited SKF38393-mediated responses at 0.1-10 microM concentrations, whereas greater concentrations reversed the inhibition. SKF38393 effects were completely blocked by the DG kinase inhibitor R59022, thus precluding any role for phospholipase-D or de novo phosphatidate synthesis in the dopaminergic response. D609 which inhibits phosphatidylcholine-specific phospholipase-C (PLC), potently inhibited both CDP-DG accumulation and inositol phosphate formation. These findings demonstrate that the selective D1-receptor antagonist SCH23390 is a partial agonist at the D1-like dopamine receptor that couples to phosphoinositide signaling, that dopaminergic facilitation of phosphoinositide signaling is independent of de novo phosphatidate synthesis, and that the widely used enzyme inhibitor, D-609, is probably not selective for phosphatidylcholine-specific PLC in brain slice preparations. The greater sensitivity of the CDP-DG measurement presents this assay as a reliable and possibly superior index of dopamine receptor-coupled PLC activation in intact tissues.

BACKGROUND: Black men in the United States have substantially higher prostate cancer incidence rates than the general population. The extent to which this incidence disparity is because prostate cancer is more prevalent, more aggressive, and/or more frequently diagnosed in black men is unknown.METHODS: The authors estimated 3 independently developed models of prostate cancer natural history in black men and in the general population using an updated reconstruction of prostate-specific antigen screening, based on the National Health Interview Survey in 2005 and on prostate cancer incidence data from the Surveillance, Epidemiology, and End Results program during 1975 through 2000. By using the estimated models, the natural history of prostate cancer was compared between black men and the general population.RESULTS: The models projected that from 30% to 43% (range across models) of black men develop preclinical prostate cancer by age 85 years, a risk that is (relatively) 28% to 56% higher than that in the general population. Among men who had preclinical disease onset, black men had a similar risk of diagnosis (range, 35%-49%) compared with the general population (32%-44%), but their risk of progression to metastatic disease by the time of diagnosis was from 44% to 75% higher than that in the general population.CONCLUSIONS: Prostate cancer incidence patterns implicate higher incidence of preclinical disease and higher risk of metastatic progression among black men. The findings suggest screening black men earlier than white men and support further research into the benefit-harm tradeoffs of more aggressive screening policies for black men. Cancer 2017;123:2312-2319. © 2017 American Cancer Society.

The imitation switch (ISWI) complex from yeast containing the Isw2 and Itc1 proteins was shown to preferentially slide mononucleosomes with as little as 23 bp of linker DNA from the end to the center of DNA. The contacts of unique residues in the histone fold regions of H4, H2B, and H2A with DNA were determined with base pair resolution before and after chromatin remodeling by a site-specific photochemical cross-linking approach. The path of DNA and the conformation of the histone octamer in the nucleosome remodeled or slid by ISW2 were not altered, because after adjustment for the new translational position, the DNA contacts at specific sites in the histone octamer had not been changed. Maintenance of the canonical nucleosome structure after sliding was also demonstrated by DNA photoaffinity labeling of histone proteins at specific sites within the DNA template. In addition, nucleosomal DNA does not become more accessible during ISW2 remodeling, as assayed by restriction endonuclease cutting. ISW2 was also shown to have the novel capability of counteracting transcriptional activators by sliding nucleosomes through Gal4-VP16 bound initially to linker DNA and displacing the activator from DNA.

UNLABELLED: A requirement for prolonged ventilation (>28 days) has been associated with a poor outcome in infants. We postulated that in the present population of infants who usually receive antenatal steroids and post-natal surfactant, prolonged ventilation in discrete episodes, i.e. discontinuous intermittent positive pressure ventilation (IPPV), would have a better outcome than a requirement for prolonged ventilation continuously from birth (continuous IPPV) and, in addition, that an abnormal ultrasound scan appearance would be a reliable predictor of poor outcome in infants requiring prolonged ventilation. All very low birth weight (VLBW) infants ventilated for at least 28 days (prolonged ventilation) were identified from a prospectively maintained database. At 1 year of age, neurodevelopmental status was assessed and abnormal neurodevelopmental outcome diagnosed if the infant's Griffiths developmental quotient was at least two standard deviations below the mean and/or they had impairment with disability. Of 417 VLBW infants, 41 required prolonged ventilation (30 continuous and 11 discontinuous). In the continuous IPPV group, 18 and one in the discontinuous IPPV group died or had abnormal neurodevelopmental outcome ( P<0.01). All eight infants with major cranial ultrasound abnormalities died or had abnormal outcome ( P<0.01).CONCLUSION: prolonged ventilation can be associated with intact survival, but not in very low birth weight infants with evidence of significant brain injury.

Gliquidone (Glurenorm) and glibenclamide were compared in a cross-over study of 20 non insulin dependent diabetics. Both drugs achieved similar levels of control, as measured by self home monitoring, glycosylated haemoglobin and N-acetyl-beta-D-glucosamidase levels. However, with glibenclamide hyperglycaemic control was only achieved at the expense of more hypoglycaemic episodes. The study examined the tailoring of drug dosage to the patients needs and found that in the majority of patients it was necessary to give both glibenclamide and gliquidone thrice daily. It confirmed the acceptability of self glucose monitoring in a home population.

Heme-derived linear tetrapyrroles (phytobilins) in phycobiliproteins and phytochromes perform critical light-harvesting and light-sensing roles in oxygenic photosynthetic organisms. A key enzyme in their biogenesis, phycocyanobilin:ferredoxin oxidoreductase (PcyA), catalyzes the overall four-electron reduction of biliverdin IXalpha to phycocyanobilin--the common chromophore precursor for both classes of biliproteins. This interconversion occurs via semireduced bilin radical intermediates that are profoundly stabilized by selected mutations of two critical catalytic residues, Asp105 and His88. To understand the structural basis for this stabilization and to gain insight into the overall catalytic mechanism, we report the high-resolution crystal structures of substrate-loaded Asp105Asn and His88Gln mutants of Synechocystis sp. PCC 6803 PcyA in the initial oxidized and one-electron reduced radical states. Unlike wild-type PcyA, both mutants possess a bilin-interacting axial water molecule that is ejected from the active site upon formation of the enzyme-bound neutral radical complex. Structural studies of both mutants also show that the side chain of Glu76 is unfavorably located for D-ring vinyl reduction. On the basis of these structures and companion (15)N-(1)H long-range HMQC NMR analyses to assess the protonation state of histidine residues, we propose a new mechanistic scheme for PcyA-mediated reduction of both vinyl groups of biliverdin wherein an axial water molecule, which prematurely binds and ejects from both mutants upon one electron reduction, is required for catalytic turnover of the semireduced state.

This article focuses on the increasing importance of effective communication between scientists and their clinical colleagues. Some recent and innovative programs to facilitate these interactions are also discussed.

The current system of International Standards (ISO) is assessed to consider whether standards are fit for purpose for the future in the context of climate change. ISO 7243, ISO 7933 and ISO 9886 provide the current ISO system for the assessment of heat stress. These involve a simple monitoring index, an analytical approach and physiological monitoring, respectively. The system relies on accurate measurement of the thermal conditions experienced by the worker (ISO 7726); and estimations of metabolic heat production due to work (ISO 8996) and the thermal properties of clothing (ISO 9920). As well as standards for heat stress assessment, the full range of ISO standards and the physical environment is listed as well as current work and proposed standards. A particular 'gap' in anticipating requirements for ISO standards in the future is the link between meteorological data and ISO standards. This is important for predicting the global consequences of a changing climate and anticipating potential impacts on occupational health across countries and cultures.

Prenatal sonographic diagnosis of an unusual case of a large left congenital diaphragmatic hernia is presented. The fetal stomach was located within the right thorax, posterior to the dextropositioned heart. Color and pulsed Doppler ultrasonography documented intrathoracic umbilical vein and ductus venosus vessels and respective waveforms, confirming extensive herniation of the left hepatic lobe.

OBJECTIVE: The purpose of this study was to measure the dose reduction achieved with dynamically adjustable z-axis collimation.MATERIALS AND METHODS: A commercial CT system was used to acquire CT scans with and without dynamic z-axis collimation. Dose reduction was measured as a function of pitch, scan length, and position for total incident radiation in air at isocenter, accumulated dose to the center of the scan volume, and accumulated dose to a point at varying distances from a scan volume of fixed length. Image noise was measured at the beginning and center of the scan.RESULTS: The reduction in total incident radiation in air at isocenter varied between 27% and 3% (pitch, 0.5) and 46% and 8% (pitch, 1.5) for scan lengths of 20 and 500 mm, respectively. Reductions in accumulated dose to the center of the scan were 15% and 29% for pitches of 0.5 and 1.5 for 20-mm scans. For scan lengths greater than 300 mm, dose savings were less than 3% for all pitches. Dose reductions 80 mm or farther from a 100-mm scan range were 15% and 40% for pitches of 0.5 and 1.5. With dynamic z-axis collimation, noise at the extremes of a helical scan was unchanged relative to noise at the center. Estimated reductions in effective dose were 16% (0.4 mSv) for the head, 10% (0.8 and 1.4 mSv) for the chest and liver, 6% (0.8 mSv) for the abdomen and pelvis, and 4% (0.4 mSv) and 55% (1.0 mSv) for coronary CT angiography at pitches of 0.2 and 3.4.CONCLUSION: Use of dynamic z-axis collimation reduces dose in helical CT by minimizing overscanning. Percentage dose reductions are larger for shorter scan lengths and greater pitch values.

The protein spectrum of blood serum was studied in healthy cows and those suffering from leucosis. The total amount of protein at the early stages of the disease is established to be unchanged. But with lympholeucosis the content of the globulin fraction increases with a simultaneous decrease in the amount of albumins within 20%. The globulin fraction content rises due to immunoglobulin G. The albumin: globulins coefficient is 0.56 for animals with leucosis and 0.84 for the healthy ones. A problem on possible qualitative changes in this fraction similar to those occurring with other forms of malignant growth is under discussion.

Tanshinone IIA (Tan-IIA), one of the major lipophilic components isolated from the root of Salviae Miltiorrhizae, has been found to exhibit anticancer activity in various cancer cells. We have demonstrated that Tan-IIA induces apoptosis in several human cancer cells through caspase- and mitochondria-dependent pathways. Here we explored the anticancer effect of Tan-IIA in human bladder cancer cell lines. Our results showed that Tan-IIA caused bladder cancer cell death in a time- and dose-dependent manner. Tan-IIA induced apoptosis through the mitochondria-dependent pathway in these bladder cancer cells. Tan-IIA also suppressed the migration of bladder cancer cells as revealed by the wound healing and transwell assays. Finally, combination therapy of Tan-IIA with a lower dose of cisplatin successfully killed bladder cancer cells, suggesting that Tan-IIA can serve as a potential anti-cancer agent in bladder cancer.

An alcoholic with no history of clinical pancreatitis was found to have pancreas divisum and marked changes of chronic pancreatitis isolated to the ventral pancreas. Pancreas divisum has been suggested to cause recurrent pancreatitis in some patients. Gross and histologic changes of pancreatitis in only the dorsal pancreas of surgically resected specimens from patients with pancreas divisum is thought to support the concept that obstruction at the minor papilla produces dorsal pancreatitis. Alternative explanations for the occurrence of segmental pancreatitis and the possible synergistic role of ethanol and bile are reviewed.

PURPOSE: We aimed to determine the feasibility of targeting low-normal or high-normal mean arterial pressure (MAP) after out-of-hospital cardiac arrest (OHCA) and its effect on markers of neurological injury.METHODS: In the Carbon dioxide, Oxygen and Mean arterial pressure After Cardiac Arrest and REsuscitation (COMACARE) trial, we used a 23 factorial design to randomly assign patients after OHCA and resuscitation to low-normal or high-normal levels of arterial carbon dioxide tension, to normoxia or moderate hyperoxia, and to low-normal or high-normal MAP. In this paper we report the results of the low-normal (65-75 mmHg) vs. high-normal (80-100 mmHg) MAP comparison. The primary outcome was the serum concentration of neuron-specific enolase (NSE) at 48 h after cardiac arrest. The feasibility outcome was the difference in MAP between the groups. Secondary outcomes included S100B protein and cardiac troponin (TnT) concentrations, electroencephalography (EEG) findings, cerebral oxygenation and neurological outcome at 6 months after cardiac arrest.RESULTS: We recruited 123 patients and included 120 in the final analysis. We found a clear separation in MAP between the groups (p < 0.001). The median (interquartile range) NSE concentration at 48 h was 20.6 µg/L (15.2-34.9 µg/L) in the low-normal MAP group and 22.0 µg/L (13.6-30.9 µg/L) in the high-normal MAP group, p = 0.522. We found no differences in the secondary outcomes.CONCLUSIONS: Targeting a specific range of MAP was feasible during post-resuscitation intensive care. However, the blood pressure level did not affect the NSE concentration at 48 h after cardiac arrest, nor any secondary outcomes.

Two galactosyltransferases were identified in human kidney microsomes which both transfer galactose from UDP Gal to lactose as well as to lactosylceramide. Using a solubilized and a partially purified enzyme preparation sufficient product could be obtained for detailed structural analysis. The trisaccharide products were isolated by gel permeation chromatography and separated by preparative high performance thin layer chromatography. The anomeric configuration of the transferred galactose was determined by specific glycosidase digestion and the linkage was identified by methylation and gas-liquid-chromatography. The glycolipid products were not separated but analyzed directly, before and after alpha or beta galactosidase digestion, by methylation, hydrolysis and thin layer chromatography. Into both acceptor substrates galactose was incorporated in alpha 1-4 (30%) and beta 1-3 (70%) linkages. The alpha 1-4 galactosyltransferase is responsible for the synthesis of the Pk antigen Gal alpha 1-4 Gal beta 1-4 Glc-ceramide in human kidney. The beta 1-3 galactosyltransferase has not previously been identified.

We have investigated the ability of metastatic cells to produce the macrophage cytokine, TNF-alpha/cachectin, as these cells have macrophage-like properties such as infiltration and migration. We looked for TNF-alpha/cachectin in three tumor cell lines derived from human malignant melanomas and six co-cultivated malignant melanomas derived, in vitro, from these three cell lines plus angioma fibroblasts. Immunohistochemistry with an anti-TNF-alpha/cachectin monoclonal antibody showed that TNF-alpha/cachectin was produced by two of the three parent melanoma cell lines. All the tumor cells in both the co-cultivated malignant melanomas and their in vitro tumorous nodules produced TNF-alpha/cachectin, even those derived from the melanoma cell line, which originally did not. The results clearly show that TNF-alpha/cachectin can be produced by non-hematopoietic tumor cells. A co-cultivated tumor model prepared from other types of human tumor cell lines promises to provide a useful tool for exploring the relationship between TNF-alpha/cachectin and oncogenesis.

INTRODUCTION: Cyclophilin A (CypA) is implicated in rheumatoid arthritis (RA) pathogenesis. We studied whether a novel anti-CypA single domain antibody (sdAb) treatment would modulate the severity of the disease in two different animal models of RA.METHODS: A novel sdAb, named sdAbA1, was screened from an immunized camel sdAb library and found to have a high binding affinity (KD = 6.9 ? 10-9 M) for CypA. The SCID-HuRAg model and the collagen-induced arthritis (CIA) in mice were used to evaluate the effects of sdAbA1 treatment on inflammation and joint destruction. For in vitro analysis, monocytes/macrophages were purified from synovial fluid and peripheral blood of patients with RA and were tested for the effect of anti-CypA sdAb on metalloproteinase (MMP) production. Human monocyte cell line THP-1 cells were selected and western blot analyses were performed to examine the potential signaling pathways.RESULTS: In the CIA model of RA, the sdAbA1 treatment resulted in a significant decrease in clinical symptoms as well as of joint damage (P <0.05). In the SCID-HuRAg model, treatment with anti-CypA antibody sdAbA1 significantly reduced cartilage erosion, inflammatory cell numbers and MMP-9 production in the implanted tissues (P <0.05). It also significantly reduced the levels of human inflammatory cytokines IL-6 and IL-8 in mouse serum (P <0.05). No toxic effects were observed in the two animal models. In vitro results showed that sdAbA1 could counteract CypA-dependent MMP-9 secretion and IL-8 production by interfering with the ERK-NF-êB pathway.CONCLUSIONS: Blockade of CypA significantly inhibited synovitis and cartilage/bone erosion in the two tested animal models of RA. Our findings provide evidence that sdAbA1 may be a potential therapeutic agent for RA.

Complex formation among transforming growth factor-â (TGF-â) receptors and its modulation by coreceptors represent an important level of regulation for TGF-â signaling. Oligomerization of ALK5 and the type II TGF-â receptor (TâRII) has been thoroughly investigated, both in vitro and in intact cells. However, such studies, especially in live cells, are missing for the endothelial cell coreceptor endoglin and for the ALK1 type I receptor, which enables endothelial cells to respond to TGF-â by activation of both Smad2/3 and Smad1/5/8. Here we combined immunoglobulin G-mediated immobilization of one cell-surface receptor with lateral mobility studies of a coexpressed receptor by fluorescence recovery after photobleaching (FRAP) to demonstrate that endoglin forms stable homodimers that function as a scaffold for binding TâRII, ALK5, and ALK1. ALK1 and ALK5 bind to endoglin with differential dependence on TâRII, which plays a major role in recruiting ALK5 to the complex. Signaling data indicate a role for the quaternary receptor complex in regulating the balance between TGF-â signaling to Smad1/5/8 and to Smad2/3.

BACKGROUND: In the early stages of human placentation, the decidua is invaded by fetal extravillous trophoblast (EVT) cells. Interactions between EVT cells and local decidual leukocytes are likely to contribute to immunological accommodation of the semi-allogeneic fetus.METHODS AND RESULTS: Natural-killer group 2 member D (NKG2D) and 2B4 (CD244) are receptors ubiquitously expressed by the distinctive population of CD56 bright, uterine natural-killer cells, which dominate the decidua at the time of implantation. Here, we investigate the UL-16 binding protein (ULBP) and MHC class-I chain related molecule (MIC) ligands of NKG2D, the CD48 ligand of 2B4 and the non-classical HLA class-I molecule, HLA-F, at the maternal-fetal interface of normal pregnancies. For many of these molecules, significant mRNA expression was detected by RT-PCR in decidual and placental tissue throughout gestation. Flow cytometry of isolated cells or immunohistological staining of implantation site sections was then performed. No protein expression of NKG2D ligands or HLA-F could be detected in decidual leukocytes or fetal trophoblast cells from the first trimester. An NKG2D-Fc fusion protein identified no novel ligands for this promiscuous receptor at the maternal-fetal interface. Strong surface protein expression of CD48 by decidual leukocytes but not by trophoblast cells was detected by flow cytometry. Histological staining showed a clear aggregation of CD48(+) cells around transformed spiral arteries of the implantation site.CONCLUSIONS: We conclude that the role of NKG2D and 2B4 is not focussed on trophoblast recognition in normal pregnancy, but is more likely involved in cross-talk among maternal cells of the placental bed.

A series of palladium(II) (1-3) and platinum(II) chloride complexes (4 and 5) with 2,2':6',2″-terpyridine (terpy) derivatives substituted at the 4' position, was synthesized and fully characterized. Single crystal X-ray diffraction analysis of complexes 2, 3 and 5 showed tridentate coordination of the 4'-substituted terpyridine (terpy) ligands to the metal center. Moreover, in vitro cytotoxic activity of these complexes toward a panel of human cancer cell lines (lung cancer A549, colorectal cancer HCT116, ovarian cancer IGROV-1) and toward normal cell line HDF (dermal fibroblast) was determined by Trypan Blue exclusion assay. Overall, the tested compounds manifested a relevant cytotoxicity for the selected cancer cell lines with complex 4 also showing a modest cytotoxicity on the normal cell lines. To better understand the mode of action of these metal complexes, their reactivity with three model proteins, i.e. hen egg white lysozyme (HEWL), cytochrome c (cyt c) and ribonuclease A (RNase A) were comparatively investigated through ESI-MS analysis. The results highlighted a different behavior between the two series of complexes being platinum compounds more reactive toward RNase and cyt c than palladium compounds. Based on the obtained results, it is proposed that in presence of RNase A and cyt c, the platinum complexes undergo activation through release of labile ligands followed by binding to the protein. In contrast, palladium complexes revealed a far lower reactivity implying the likely occurrence of a different mechanism of action.

In onchocerciasis patients and in O. volvulus-exposed individuals without signs of onchocericiasis, T- and B-cell responses to two recombinantly expressed O. volvulus enzymes were analysed and compared to responses to total protein extract of adult parasites. The cytosolic enzymes Cu/Zn superoxide dismutase 1 (OvSOD1) and glutathione S-transferase 2 (OvGST2) represent 2 detoxifying molecules which may play an important role in parasite defense against host-induced oxidative stress. The T-cell response to the two recombinant proteins was analysed by investigating the cytokine responses of peripheral blood mononuclear cells. Induction of IL-5 at the mRNA level and IL-5 and IL-10 at the protein level was demonstrated in patients with the generalized form of onchocerciasis and endemic normals without clinical manifestations. IFN-gamma was not found to be induced by either antigen. This pattern of lymphokine expression is indicative of a Th2-type response. Compared to patients with the generalized form, a higher level of cytokine induction was observed in the group of endemic normals. Low but significant IgG levels were observed against OvSOD1 in patients with onchocerciasis; higher antibody levels were found against OvGST2 in patients and endemic normals. The highest IgG levels were detected against the crude O. volvulus extract. These results indicate that the two recombinant O. volvulus proteins induce moderate T and B cell responses.

BACKGROUND: De novo development of anti-human leukocyte antigen (HLA) antibodies after transplantation is associated with increased rejection and decreased graft survival. In this study, the effect of posttransplant HLA antibodies on clinical outcome was evaluated in patients treated with tacrolimus by means of flow cytometric crossmatches (FCXm).METHODS: T- and B-cell FCXm were performed retrospectively on posttransplant sera of patients who received a graft between 1997 and 1999. Ninety-four kidney-only recipients were tested and all FCXm positive sera were investigated for the presence of HLA class I and II antibodies by Flow panel reactive antibodies.RESULTS: From 94 patients with a negative pretransplant complement-dependent cytotoxicity crossmatch, seven (7%) showed a positive pretransplant FCXm. After transplantation the FCXm became positive in five patients (6%). The predictive value of a positive FCXm after transplantation, and the log-transformed relative change in fluorescence ratio between pretransplant and posttransplant serum, were not significant to rejection within six months, nor to graft survival censored for death.CONCLUSIONS: The presence of HLA antibodies before rejection or graft failure could only be shown in a minority of patients; most antibodies were detected after graft failure, especially after transplantectomy. Monitoring through antibody testing after transplantation on the basis of our results has no added value with tacrolimus-based immunosuppression.

Transglutaminase 1 (TGase 1) is a Ca(2+)-dependent enzyme which catalyzes epsilon-(gamma-glutamyl)lysine cross-linking of substrate proteins such as involucrin and loricrin to generate the cornified envelope at the cell periphery of the stratum corneum. We have shown that disruption of the TGase 1 gene in mice results in neonatal lethality, absence of the cornified envelope, and impaired skin barrier function. Based on the importance of TGase 1 in epidermal morphogenesis, we have now assessed its role in wound healing. In neonatal mouse skin, TGase 1 mRNA as well as keratin 6alpha was induced in the epidermis at the wound edges as early as 2 hours after injury and that expression continued in the migrating epidermis until completion of re-epithelialization. The TGase 1 enzyme co-localized on the plasma membrane of migrating keratinocytes with involucrin, but not with loricrin, which suggests the premature assembly of the cornified envelope. Similar injuries to TGase 1 knockout mouse skins grafted on athymic nude mice showed substantial delays in wound healing concomitant with sustained K6alpha mRNA induction. From these results, we suggest that activation of the TGase 1gene is essential for facilitated repair of skin injury.

Transmitter release at synapses between nerve cells is spatially restricted to active zones, where synaptic vesicle docking, priming, and Ca2+-dependent fusion take place in a temporally highly coordinated manner. Munc13s are essential for priming synaptic vesicles to a fusion competent state, and their specific active zone localization contributes to the active zone restriction of transmitter release and the speed of excitation-secretion coupling. However, the molecular mechanism of the active zone recruitment of Munc13s is not known. We show here that the active zone recruitment of Munc13 isoforms Munc13-1 and ubMunc13-2 is regulated by their binding to the Rab3A-interacting molecule RIM1alpha, a key determinant of long term potentiation of synaptic transmission at mossy fiber synapses in the hippocampus. We identify a single point mutation in Munc13-1 and ubMunc13-2 (I121N) that, depending on the type of assay used, strongly perturbs or abolishes RIM1alpha binding in vitro and in cultured fibroblasts, and we demonstrate that RIM1alpha binding-deficient ubMunc13-2(I121) is not efficiently recruited to synapses. Moreover, the levels of Munc13-1 and ubMunc13-2 levels are decreased in RIM1alpha-deficient brain, and Munc13-1 is not properly enriched at active zones of mossy fiber terminals of the mouse hippocampus if RIM1alpha is absent. We conclude that one function of the Munc13/RIM1alpha interaction is the active zone recruitment of Munc13-1 and ubMunc13-2.

We performed bedside testing for peripheral neuropathy in our systemic sclerosis (SSc) population to determine whether foot care guidelines should be developed for SSc. Twenty consecutive SSc patients and 20 healthy control (HC) patients were evaluated for peripheral neuropathy in both feet using the 10-g Semmes-Weinstein monofilament examination (SWME) and 128 Hz vibration sensation using the on-off method. Independent, blinded, vibratory sensation, and SWME evaluations were performed on each subject by two investigators who had completed a training session to standardize each exam. An additional consecutive 20 patients with type 2 diabetes mellitus (DM) were examined by a diabetologist to compare with peripheral neuropathy prevalence in SSc patients. We examined the inter-rater variability using Cohen's kappa. We compared SWME and vibratory sensation in SSc to HC using Fisher's exact. The t test was used to compare duration of disease and modified Rodnan skin score (mRSS) for those with abnormal SWME or vibratory sensation. Two of 20 SSc patients reported sensory foot symptoms consistent with peripheral neuropathy prior to the examination. Inter-rater agreement for both SWME and vibratory sensation was strong (kappa: 0.72 and 0.83, respectively). Two HC and 12 SSc patients demonstrated abnormal vibratory sense (one-sided Fishers' exact, p < 0.002). No HC and four SSc patients had abnormal monofilament exams (one-sided Fisher's exact, p = 0.053). Neither mRSS (p = 0.28) nor duration of non-Raynauds (p = 0.07) symptoms differed between those with peripheral neuropathy and those without. Duration of Raynaud's symptoms were clinically significantly associated with presence of peripheral neuropathy (p = 0.04). The prevalence of sensory loss to monofilament in SSc was identical to DM patients (4/20). SSc patients have a considerable prevalence of pedal peripheral neuropathy as detected by loss of vibratory sensation or inability to sense the 10-g SWME. Further studies are indicated to determine if routine screening for neuropathy and subsequent podiatric care for SSc patients with abnormalities can reduce pedal complications.

A series of fluorescent teraryls and quateraryls were prepared from a ketene-S,S-acetal under mild conditions. These compounds exhibited blue, sky-blue and green color emissions both in the solid state and in a solution with good quantum yields, positive solvatochromic behavior, and reversible oxidation and reduction properties. The electronic characteristics of teraryl 6a and quateraryls 9a,b were examined by time-dependent density functional theory (TDDFT) calculations. Light-emitting devices were fabricated from teraryl 6a and quateraryls 9a,b as dyes and the configuration of ITO/PEDOT:PSS (40 nm)/NPB (20 nm)/ dye (50 nm)/BCP (7 nm)/ LiF (0.7 nm)/Al (200 nm), which exhibited electroluminescence maxima of 455, 480, and 525 nm, respectively. These devices operated at a substantially low turn-on voltage (3 and 4 V) and exhibited maximum luminance efficiencies of 0.62, 0.57, and 1.9 cd/A and brightnesses of 59, 160, and 1284 cd/m(2), respectively.

OBJECTIVES: We aimed to compare the performance of different regression modeling approaches for the prediction of heterogeneous treatment effects.STUDY DESIGN AND SETTING: We simulated trial samples (n = 3,600; 80% power for a treatment odds ratio of 0.8) from a superpopulation (N = 1,000,000) with 12 binary risk predictors, both without and with six true treatment interactions. We assessed predictions of treatment benefit for four regression models: a "risk model" (with a constant effect of treatment assignment) and three "effect models" (including interactions of risk predictors with treatment assignment). Three novel performance measures were evaluated: calibration for benefit (i.e., observed vs. predicted risk difference in treated vs. untreated), discrimination for benefit, and prediction error for benefit.RESULTS: The risk modeling approach was well-calibrated for benefit, whereas effect models were consistently overfit, even with doubled sample sizes. Penalized regression reduced miscalibration of the effect models considerably. In terms of discrimination and prediction error, the risk modeling approach was superior in the absence of true treatment effect interactions, whereas penalized regression was optimal in the presence of true treatment interactions.CONCLUSION: A risk modeling approach yields models consistently well calibrated for benefit. Effect modeling may improve discrimination for benefit in the presence of true interactions but is prone to overfitting. Hence, effect models-including only plausible interactions-should be fitted using penalized regression.

The study investigated the effect of colistin exposure on calcium homeostasis and mitochondria functions in neurons. We used an in vitro drug model to induce neurotoxicity that closely mimic the in vivo condition by exposing primary cultures of chick cortex neurons to different concentrations of 0, 0.83, 4.15 and 8.3 ìg/mL colistin. The cell activity was determined by methods of MTT and lactate dehydrogenase release at 24 h post-beginning. The membrane potential (ÄØm) and ultrastructure of mitochondrial were assessed. The calcium ion concentration within neurons ([Ca(2+)]i) was detected using the Fura3/AM as the probe and expression level of intracellular calmodulin (CaM) mRNA was detected by reverse transcription polymerase chain reaction. The results showed that, in the 4.15 and 8.3 ìg/mL colistin groups, the functions of mitochondria altered, the ÄØm was significantly decreased and the mitochondrial cristae was swollen and even vacuolar degeneration in mitochondria occurred. Moreover, the expression level of colistin could decrease CaM mRNA, and increase free calcium concentration. The present work revealed that colistin-induced mitochondria dysfunction and calcium homeostasis disequilibrium, providing new insight into the toxicological mechanism of colistin in neurons.

AIM: To assess the prognostic significance of cathepsin L, a cysteine protease that degrades the peri-tumoral tissue, in patients with pancreatic cancer.METHODS: Plasma samples from 127 pancreatic cancer patients were analyzed for cathepsin L levels by ELISA. Out of these patients, 25 underwent surgery and their paraffin-embedded tissue was analyzed for cathepsin L expression by immunohistochemistry. Survival of patients and clinicopathological parameters was correlated with cathepsin L expression in plasma and tissue using appropriate statistical analysis.RESULTS: The mean (± SD) cathepsin L in plasma samples of pancreatic cancer patients was 5.98 ± 2.5 ng/mL that was significantly higher compared to the levels in healthy controls (3.83 ± 0.45) or chronic pancreatitis patients (3.97 ± 1.06). Using ROC curve, a cut-off level of 5.0 ng/mL was decided for survival analysis. Elevated plasma levels of cathepsin L were found to be associated with poor prognosis (P = 0.01) in multivariate analysis. The plasma levels of the protease decreased after surgery. Though no significant correlation was seen between plasma and tissue expression of this protease, a trend did emerge that high cathepsin L expression in tissue correlated with its high levels in plasma.CONCLUSION: Cathepsin L levels in plasma of pancreatic cancer patients may be used as a potential prognostic marker for the disease.

The author investigated in patient material from 12 years 251 cases involving spontaneous abortion and delivery in the second trimester. The material included 28 sets of twins and one set of triplets, thus the total number of fetuses and neonates was 281. The sex ratio was 136.1, higher than that documented natural sex ratio in the second trimester or at term. The ratio approaches that experienced in cases of perinatal death.

The recombinant Muscovy duck parvovirus (rMDPV) has been recently characterized and identified in China. However, whether other additional rMDPV field isolates exist, and whether these strains possess common molecular characteristics, remain to be explored. In this retrospective study, two new rMDPV isolates, namely, JH06 and JH10, were identified through genome sequencing and recombination analysis. JH06, JH10, and four previously characterized rMDPV strains (SAAS-SHNH, ZW, FJM3, and PT97) underwent the same recombination events in a 1.1-kb region in their VP3 genes and displayed highly consistent beginning and ending breakpoints. JH06, JH10, SAAS-SHNH, ZW, and FJM3, but not PT97, underwent recombination in their P9 promoter regions. In both recombination events, the classical MDPV strain YY acted as the major parent, whereas the virulent strain DY16 and the vaccine strain SYG61v of goose parvovirus (GPV) served as the minor parents. The sequence alignments of inverted terminal repeats (ITRs) revealed that rMDPV strains shared higher identities (96.0%-97.2%) with classical MDPV strains than with GPV and contained typical one-nucleotide-pair deletions in the palindromic stems of their ITRs. This work elucidated the common molecular characteristics and differences of six rMDPV strains. The results of this work will facilitate the preparation of an efficacious vaccine for the protection of Muscovy ducks against rMDPV infection.

Arterial pressure fluctuates rhythmically in healthy supine resting humans, who, from all outward appearances, are in a 'steady-state'. Others have asked, If baroreflex mechanisms are functioning normally, how can arterial pressure be so variable? We reanalysed data from nine healthy young adult men and women and tested the hypotheses that during brief periods of observation, human baroreflex sensitivity fluctuates widely and rhythmically. We estimated vagal baroreflex sensitivity with systolic pressure and R-R interval cross-spectra measured over 15 s segments, moved by 2 s steps through 20-min periods of frequency- and tidal volume-controlled breathing. We studied each subject at the same time on three separate days, with fixed protocols that included two physiological states, supine and passive 40 deg upright tilt, before and after beta-adrenergic, cholinergic, and angiotensin converting enzyme blockade. Minimum, mean and maximum (+/-s.d.) supine control baroreflex sensitivities averaged 5 +/- 3, 18 +/- 6, and 55 +/- 22 ms mmHg(-1). In most subjects, moderate ongoing fluctuations of baroreflex sensitivity were punctuated by brief major peaks, yielding frequency distributions that were skewed positively. Fast Fourier transforms indicated that baroreflex sensitivity fluctuations (expressed as percentages of total power) concentrated more in very low, 0.003-0.04 Hz, than ultra low, 0.0-0.003 Hz, frequencies (77 +/- 7 versus 11 +/- 8%, P < or = 0.001, rank sum test). Autoregressive centre frequencies averaged 0.012 +/- 0.003 Hz. The periodicity of very low frequency baroreflex sensitivity fluctuations was not influenced significantly by upright tilt, or by variations of autonomic drive or angiotensin activity. Our analysis indicates that during ostensibly 'steady-state' conditions, human vagal baroreflex sensitivity fluctuates in a major way, at very low frequencies.

Based on the oxidation hypothesis high doses of alpha-tocopherol have been advocated to prevent atherosclerosis, but clinical trials failed to demonstrate a benefit. As specific oxylipids activate PPARgamma and LXRalpha, master regulators of lipid metabolism and cholesterol exporters, we hypothesized, that high dose alpha-tocopherol might interfere with reverse cholesterol transport out of the vessel wall. Human THP-1 cells, a foam cell model, were preincubated with alpha-tocopherol or carrier before exposure to oxidized LDL, delipidated HDL or control buffer. Specific mRNAs were quantified by real-time RT-PCR, LXRalpha activation by a reporter gene assay and cellular cholesterol homeostasis by oxLDL and dHDL facilitated uptake and efflux assays. alpha-Tocopherol significantly reduced baseline expression and stimulation by oxLDL of LXRalpha activity, CD36, ABCA1, and ABCG1. alpha-Tocopherol also reversed the suppression of CD36 and ABCA1 by dHDL. Thus alpha-Tocopherol compromises cellular lipid scavenging and channelling of cholesterol into reverse transport out of the vessel wall.

In 2007, the Rhodophyceae Alsidium corallinum C. Ag., a marine taxon, bloomed in the eutrophic lagoon of Orbetello (Tuscany, Italy) for the first time, becoming the dominant species in spring and summer. In November, its biomass collapsed. The hypothesis examined in this study is that the bloom expressed a relatively low eutrophic level of the ecosystem after intense disposal of accumulated sedimentary organic matter (OM) by dystrophic processes in the two years preceding the bloom. To verify the hypothesis, we compared water physical-chemical variables, sediment redox (Eh) and OM, and standing crops of macroalgae and seagrass from the database of routine monitoring between 2005 and 2008. We also used dissolved nutrient data obtained in 2007 and 2008, as well as data on chlorophyll and total suspended matter in the water column during the microalgal bloom of 2007, and C, N and P content in thalli of the Chlorophycea Chaetomorpha linum and the Rhodophyceae Gracilariopsis longissima and A. corallinum obtained in 2007. In 2007, unusually low values of dissolved inorganic nitrogen (DIN) were recorded. Combined with stable values of soluble reactive phosphorus (SRPs), low DIN led to a reduction of about one order of magnitude in the DIN:SRP atomic ratio with respect to the past and to 2008. G. longissima accumulated C, N and P more than the other species and A. corallinum proved to be less demanding. Sediment OM was lower in the autumn of years characterized by dystrophy, confirming that summer dystrophic events coincided with maximum energy dissipation in this ecosystem. However, as soon as OM and DIN values increased (2008), the vegetation shifted towards blooms of G. longissima and C. linum, while A. corallinum almost disappeared. The results sustain the hypothesis that the bloom of A. corallinum was due to a decline in DIN that limited G. longissima, and to intense turbidity of the water caused by microphytes that developed after the dystrophic event of summer 2006. The latter probably limited the development of C. linum, which could only develop at the edges of the lagoon.

The laparoscopic adjustable gastric banding (LAGB) procedure has been used in bariatric surgery for over 20 years, and despite its initial success, it has been most often criticized for causing adverse effects, lacking efficacy, and frequently requiring revision. Evidence-based medicine supports these criticisms, and the LAGB is no longer considered by most bariatric surgeons worldwide to be a standard operation. While we have to admit that its associated food tolerance issues and variable efficacy make the LAGB less desirable for most patients, time will tell whether the current armamentarium of bariatric procedures are still too aggressive and new safer procedures still need to be developed.

BACKGROUND/AIMS: Since pharmacotherapy of portal hypertension has always been a subject of wide interest, we decided to study the effects of different angiotensin-converting enzyme inhibitors and endoscopic sclerotherapy on portal hemodynamics in patients with portal hypertension and bleeding esophageal varices.METHODOLOGY: The study included 72 patients with portal hypertension divided into 6 equal groups. Endoscopic sclerotherapy was done to all patients every 2 weeks for 3 months. In addition, the first 5 groups of patients were maintained on angiotensin-converting enzyme inhibitors for 3 months as follows: group I on perindopril, II on ramipril, III on fosinopril, IV on lisinopril and V on captopril. Portal hemodynamics were determined before and after therapy (using an ultrasonic duplex system). New Doppler portal indices were derived and portal vein kinetic pressure was estimated for the first time by using data derived from the ultrasonic duplex system.RESULTS: 1) Short-term endoscopic sclerotherapy alone resulted in significant elevation of portal vein kinetic pressure, wall stress index and flow volume (P < 0.01) and non-significant increase in the total portal circulation resistance index (P > 0.05) and significantly decreased portal vein compliance and distensibility indices (P < 0.05); 2) Angiotensin-converting enzyme inhibitors reduced the maximum and average portal velocities, the portal flow volume, total portal circulation resistance index and increased portal vein compliance and distensibility indices, hence they reduced the portal vein kinetic pressure significantly in group IV (P < 0.05 for the flow volume and P < 0.01 for other indices); 3) The only side effect encountered was allergic cough (in 8.33% of patients). No effects were noticed on the pulse, systolic, diastolic or mean blood pressures or Child-Pugh Score of liver disease.CONCLUSIONS: 1) Angiotensin-converting enzyme inhibitors when added to endoscopic sclerotherapy can ameliorate the effects of the latter on portal hemodynamics in patients with portal hypertension; 2) Portal vein kinetic pressure, total portal circulation resistance index, portal vein wall stress index, compliance and distensibility indices are new Doppler portal indices that proved to be of value in the follow-up of patients with portal hypertension under sclerotherapy alone or in conjunction with pharmacotherapy; 3) Angiotensin-converting enzyme inhibitors are safe drugs that can be used for portal decompression with endoscopic sclerotherapy. Their use as sole portal anti-hypertensive agents still awaits further studies.

In contrast to brain, the sciatic nerve concentration of vitamin E in rats increased rapidly during the postnatal period (approximately fivefold between days 1 and 8), then decreased dramatically (about twofold between days 8 and 30), and further decreased slowly between days 30 and 60 and remained constant up to 2 years. Although the sciatic nerve concentration of vitamin E decreased by 58% between days 8 and 30, the concentration of vitamin E in serum presented a marked decrease (approximately 75%). The vitamin E concentrations varied in a similar pattern in whole sciatic nerve and in endoneurium and showed a very close correlation (r = 0.94). The age-related changes in fatty acid concentration of the endoneurial fraction of the sciatic nerve were characterized by a large increase in content of saturated and monounsaturated fatty acids up to 6 months (twofold for saturated and fourfold for monounsaturated fatty acids). Then, up to 24 months, the amount of these fatty acids decreased very slowly. The content of (n-6) polyunsaturated fatty acids (PUFAs) decreased rapidly up to 1 year and slowly afterward. In contrast, during development the amount of (n-3) PUFA was relatively stable and decreased during aging. A highly significant correlation between vitamin E and (n-6) PUFA [18:2(n-6), 20:4(n-6), and total (n-6)] was observed but not between (n-3) PUFA and vitamin E. It is suggested that there may be a relationship between vitamin E and (n-6) PUFA in the PNS membranes during development and aging.

Borna disease virus (BDV) is a nonsegmented, negative-strand RNA virus that employs several unique strategies for gene expression. The shortest transcript of BDV, X/P mRNA, encodes at least three open reading frames (ORFs): upstream ORF (uORF), X, and P in the 5' to 3' direction. The X is a negative regulator of viral polymerase activity, while the P phosphoprotein is a necessary cofactor of the polymerase complex, suggesting that the translation of X is controlled rigorously, depending on viral replication. However, the translation mechanism used by the X/P polycistronic mRNA has not been determined in detail. Here we demonstrate that the X/P mRNA autogenously regulates the translation of X via interaction with host factors. Transient transfection of cDNA clones corresponding to the X/P mRNA revealed that the X ORF is translated predominantly by uORF-termination-coupled reinitiation, the efficiency of which is upregulated by expression of P. We found that P may enhance ribosomal reinitiation at the X ORF by inhibition of the interaction of the DEAD-box RNA helicase DDX21 with the 5' untranslated region of X/P mRNA, via interference with its phosphorylation. Our results not only demonstrate a unique translational control of viral regulatory protein, but also elucidate a previously unknown mechanism of regulation of polycistronic mRNA translation using RNA helicases.

This article offers a new model for bulimia nervosa (BN) that explains both the initial impulsive nature of binge eating and purging, as well as the compulsive quality of the fully developed disorder. The model is based on a review of advances in research on BN and advances in relevant basic psychological science. It integrates transdiagnostic personality risk, eating-disorder-specific risk, reinforcement theory, cognitive neuroscience, and theory drawn from the drug addiction literature. We identify both a state-based and a trait-based risk pathway, and we then propose possible state-by-trait interaction risk processes. The state-based pathway emphasizes depletion of self-control. The trait-based pathway emphasizes transactions between the trait of negative urgency (the tendency to act rashly when distressed) and high-risk psychosocial learning. We then describe a process by which initially impulsive BN behaviors become compulsive over time, and we consider the clinical implications of our model. (PsycINFO Database Record

A series of water-soluble 2'-paclitaxel prodrugs were synthesized by attaching paclitaxel to polyethylene glycol (PEG) through amino acid spacers. The prodrugs showed highly improved water solubility, enhanced in vitro cytotoxicity and in vivo antitumor activity compared with the native drug, paclitaxel.

BACKGROUND: Visceral fat (VF) plays a major role in the development of metabolic syndrome associated with obesity. The aim of our study is to compare VF and subcutaneous fat (SCF) reduction measured by ultrasonography (US) after laparoscopic adjustable gastric banding (LAGB), laparoscopic sleeve gastrectomy (LSG), and laparoscopic Roux-En-Y gastric bypass (LRYGB).METHODS: Thirty-nine morbidly obese patients were prospectively evaluated by US before surgery and 3, 6, and 12 months following surgery to determine VF and SCF thickness.RESULTS: Three statistically comparable groups of morbidly obese patients underwent LRYGB (n = 13), LSG (n = 15), and LAGB (n = 11). The three groups did not differ in initial age, gender, body mass index (BMI), VF, or SCF. Final excess weight loss (EWL%) was highest after LSG and LRYGB followed by LAGB (81 ± 5.8 vs. 69.5 ± 4.5 vs. 43.4 ± 5.2, p < 0.001). LSG and LRYGB were significantly more efficient in VF reduction (ÄVF) compared with LAGB (7.1 ± 0.5 vs. 5.6 ± 0.6 vs. 3.6 ± 0.8, p = 0.004). SCF reduction (ÄSCF) was also highest after LSG followed by LRYGB and LAGB (3 ± 0.2 vs. 2.2 ± 0.4 vs. 1.9 ± 0.4, p = 0.08). The change in fat distribution, determined as Ä(VF/SCF), showed a preferential VF reduction in the LSG and LRYGB patients compared with patients that underwent LAGB (0.59 ± 0.1 vs. 0.52 ± 0.2 vs. 0.19 ± 0.2, p = 0.42). In a subgroup analysis comparing only LSG to LRYGB, no statistically significant difference was seen in EWL%, ÄVF, ÄSCF, or in fat distribution Ä(VF/SCF).CONCLUSION: LSG and LRYGB show better preferential and overall VF reduction than LAGB. US may serve as a simple tool of evaluating postoperative fat distribution.

Among human subjects, sleep is a time of considerable neuro-endocrine activity and the nocturnal secretion of growth hormone is intimately related to the sleep-wake cycle. In contrast, the hypothalamic-pituitary-adrenal axis of entrained individuals shows a circadian pattern of activity with secretion increasing in the latter part of sleep. A variety of neuroendocrine disturbances have been described in depressed patients who also manifest consistent disturbances in their sleep-wake cycle. The sleep-wake cycle is capable of masking these rhythms and thereby modifying patterns of hormone secretion. The sleep disturbance in depressed patients appears to have little effect on the phase of either cortisol or growth hormone secretion but does reduce their amplitudes.

BACKGROUND: Limited information is available regarding the relationship between the chemical structures and inhibitory effects of anthocyanin (ACN) on triglyceride (TG) overaccumulation. Thus this study investigated the antioxidant activity and inhibitory effect of blackberry, wild blueberry, strawberry, and chokeberry ACN-rich extracts, with different structural characteristics, on oleic acid-induced hepatic steatosis in vitro. Four major ACNs from these berries, with different aglycones, namely cyanidin-3-glucoside (Cy-3-glu), delphinidin-3-glucoside, pelargonidin-3-glucoside, and malvidin-3-glucoside, were also investigated.RESULTS: Blackberry ACN-rich extract exhibited the most significant inhibitory effect on TG clearance (30.5% ± 3.4%) and reactive oxygen species generation. TG clearance was significantly correlated with total phenolic content (r = 0.991, P < 0.05) and oxygen radical absorbance capacity value (r = 0.961, P < 0.05). Furthermore, Cy-3-glu showed the highest inhibitory effect on intracellular TG overaccumulation, with a maximum TG clearance of 61.3% at 40 µg mL(-1) .CONCLUSION: Our findings suggest that the inhibitory effects of different ACNs on oleic acid-induced hepatic steatosis significantly vary. Cy-3-glu, which contains the ortho hydroxyl group in its B ring, possibly confers the protective effects of antioxidants and inhibits TG accumulation in HepG2 cells. © 2015 Society of Chemical Industry.

A palatal lift prosthesis (PLP) is an intraoral device that provides lift for the soft palate. The usual purpose of a PLP is to reduce nasopharyngeal reflux and the hypernasal speech caused by velopharyngeal incompetence. However, for this patient, the main purpose was to relieve a functional blockage at the oropharyngeal isthmus by suspending the soft palate. A PLP with soft and flexible lift was applied in a patient with a traumatic brain injury and dysphagia. The PLP improved oropharyngeal bolus transit time by relieving the blockage at the oropharyngeal isthmus. This type of PLP may help to improve bolus transport for patients with dysphagia.

BACKGROUND: Mastication is one of the most fundamental functions for the conservation of life. The demand for devices for evaluating stomatognathic function, for instance, recording mandibular movements or masticatory muscle activities using animal models, has been increasing in recent years to elucidate neuromuscular control mechanisms of mastication and to investigate the etiology of oral motor disorders. To identify the fundamental characteristics of the jaw movements of mice, we developed a new device that reconstructs the three-dimensional (3D) movement trajectories on an arbitrary point on the mandible during mastication.METHODS: First, jaw movements with six degrees of freedom were measured using a motion capture system comprising two high-speed cameras and four reflective markers. Second, a 3D model of the mandible including the markers was created from micro-computed tomography images. Then, the jaw movement trajectory on the certain anatomical point was reproduced by integrating the kinematic data of the jaw movements with the geometric data of the mandible.RESULTS: The 3D movements at any points on the mandible, such as the condyle, molar, and incisor during mastication, could be calculated and visualized with an accuracy > 0.041 mm in 3D space. The masticatory cycle was found to be clearly divided into three phases, namely, the opening, closing, and occlusal phases in mice.CONCLUSIONS: The proposed system can reproduce and visualize the movements of internal anatomical points such as condylar points precisely by combining kinematic data with geometric data. The findings obtained from this system could facilitate our understanding of the pathogenesis of eating disorders or other oral motor disorders when we could compare the parameters of stomatognathic function of normal mice and those of genetically modified mice with oral behavioral dysfunctions.

Regional drug delivery is an approach designed to improve the selectivity of chemotherapy. When compared with systemic drug administration, regional delivery can potentially increase drug concentrations at tumor sites and/or lower systemic drug exposure. Pharmacokinetic analysis can evaluate or even predict the relative advantage of regional drug delivery. Based on a quantitative analysis of regional drug delivery, some drugs and sites of delivery are more favorable than others. The formulas for the therapeutic advantage of intraarterial, intrathecal, and intraperitoneal delivery have a similar structure. The ratio of total body clearance of a drug to its regional exchange rate is the principal determinant of therapeutic advantage. Thus, the most favorable circumstances for regional delivery are the use of drugs with high total body clearances and/or sites of delivery with low exchange rates. Pharmacodynamic factors must also be considered for the overall assessment of regional delivery.

RATIONALE: The mechanisms of action of MTX (methotrexate) in the treatment of RA (rheumatoid arthritis) and PsA (psoriatic arthritis) is related to its antifolic activity, due to the high affinity for enzymes that require folate cofactors as dihydrofolate reductase and to the anti-inflammatory activity derivated from the inhibition of thymidylate synthetase that leads to the over-production of adenosine.PATIENT CONCERNS: Our patient was a 41-year-old female, affected by PsA in treatment since 2 years with low-dose methylprednisolone and low-dose subcutaneous MTX. The treatment was effective. The patient subacutely developed a severe paraparesis with impossibility of gait or standing without aid and was admitted to a Neurology Department where the cause of the paraparesis was not clear in spite of accurate radiological neurophysiologic and laboratory tests. Therefore, she was admitted in a rehabilitation unit.DIAGNOSIS AND INTERVENTIONS: Paraparesis in PsA patient in treatment with methotrexate. MTX toxicity was hypothesized; therefore the drug was discontinued while i.m. folic acid and cyanocobalamin were administered for 20 days. The diagnosis was clinical, based on neurological examination (paraparesis) and on the chronic use of MTX (hypothesis of toxicity).OUTCOMES: The patient obtained a complete resolution of paraparesis. Genetic analyses showed associated a compound heterozygosity for the c.1298A>C and c.677C>T variants of methylenetetrahydrofolate reductase (MTHFR) gene.LESSONS: Neurological side effects of MTX are uncommon. In literature no previous case of MTX induced paraparesis in patients treated with low-dose MTX for chronic arthritis has been described. The association between the gene polymorphisms of MTHFR (c.1298A>C and c.677C>T) and MTX toxicity in arthritis patients is confirmed. The case also confirms that folates are a precious antidote of MTX toxicity.

BACKGROUND: The prognostic value of long interspersed nucleotide element-1 (LINE-1) methylation in patients with colorectal cancer (CRC) remains uncertain. We have therefore performed a meta-analysis to elucidate this issue.METHODS: The PubMed and Web of Science databases were searched for studies published up to 30 June 2016 which reported on an association between LINE-1 methylation and overall survival (OS), disease-free survival (DFS), or cancer-specific survival (CSS) among CRC patients. The reference lists of the identified studies were also analyzed to identify additional eligible studies. Hazard ratios (HRs) with 95% confidence intervals (CIs) were pooled using the fixed-effects or the random-effects model. Stratification analysis and meta-regression analysis were performed to detect the source of heterogeneity. Analyses of sensitivity and publication bias were also carried out.RESULTS: Thirteen independent studies involving 3620 CRC patients were recruited to the meta-analysis. LINE-1 hypomethylation was found to be significantly associated with shorter OS (HR 2.92, 95% CI 2.20-3.88, p < 0.001) and DFS (HR 2.18, 95% CI 1.46-3.27, p < 0.001), as well as unfavorable CSS (HR 1.96, 95% CI 1.35-2.85, p < 0.001). No heterogeneity was found among the studies evaluating the associations between LINE-1 hypomethylation and OS or DFS, with the exception being CSS. Moreover, meta-regression analysis suggested that one of the contributors to between-study heterogeneity on the association between LINE-1 methylation and CSS was statistical methodology. The subgroup analysis suggested that the association in studies using the Cox model statistical method (HR 2.76, 95% CI 1.90-4.01, p < 0.001) was stronger than that in studies using the Log-rank test (HR 1.41, 95% CI 1.07-1.87, p = 0.015).CONCLUSIONS: The results of this meta-analysis suggest that LINE-1 methylation is significantly associated with the survival of CRC patients and that it could be a predictive factor for CRC prognosis.

BACKGROUND: The association between antidepressant exposure and type 2 diabetes mellitus is still debated. Moreover, the pharmacological mechanisms remain unknown.OBJECTIVE: The objective of this study was to investigate this putative relationship with the role of antidepressant pharmacological targets using the 'pharmacoepidemiological-pharmacodynamic' method.METHODS: First, we performed case/non-case analyses in VigiBase® (the World Health Organization international database of suspected adverse drug reactions) to examine a signal of increased type 2 diabetes reporting (expressed as the reporting odds ratio and its 95% confidence interval) for antidepressants in general; examine and rank type 2 diabetes signals between the different pharmacological classes of antidepressants and the different antidepressants (58 in total). Second, we performed linear regression analyses to explore the association between the type 2 diabetes signal ranked between antidepressants and their binding affinities for nine targets (serotonin, norepinephrine, dopamine transporters, 5-HT2C serotonin, D2 dopamine, á1, á2 adrenergic, M3 muscarinic and H1 histamine receptors).RESULTS: A significant type 2 diabetes signal was found for antidepressants in general, three classes of antidepressants (tricyclic antidepressants, serotonin reuptake inhibitors and "other" antidepressants) and 15 individual antidepressants in particular. Among the antidepressants, three serotonin reuptake inhibitors [escitalopram (adjusted reporting odds ratio 1.15 [1.07-1.25]), paroxetine (1.15 [1.07-1.23]), sertraline (1.23 [1.17-1.31])] and three "other" antidepressants [duloxetine (1.15 [1.07-1.23]), trazodone (1.20 [1.09-1.32]), venlafaxine (1.15 [1.08-1.23])] were the antidepressants most frequently reported with type 2 diabetes. We found a significant correlation between the type 2 diabetes signal and serotonin transporter affinity (slope = 0.14 [0.06-0.23], p = 0.003, R2 = 0.43) but not the other targets.CONCLUSION: The present study suggests a potential role for serotonin transporter in antidepressant-induced type 2 diabetes.

The effect of various doses, viz. 0, 0.5, 1, 2, 4, 6 and 8 mg/kg body weight of naringin (NIN) (a citrus flavanone) was studied on the alteration in the radiation-induced micronucleated polychromatic (MPCE) and normochromatic (MNCE) erythrocytes in mouse bone marrow exposed to 2 Gy of 60Co gamma-radiation. The treatment of mice with various doses of NIN before exposure to 2 Gy resulted in a significant decline in the frequency of MPCE when compared to the non-drug-treated irradiated control. However, the greatest reduction in MPCE was observed for 2mg/kg body weight NIN, accompanied by a highest PCE/NCE ratio when compared with the non-drug-treated irradiated control. Therefore, further studies were carried out using this dose of NIN, where the animals were administered with 2mg/kg body weight of NIN before exposure to 0, 0.5, 1, 2, 3 and 4 Gy of gamma-radiation. The frequency of MPCE and MNCE increased in a dose-dependent manner in both the non-drug-treated irradiated control and NIN-pretreated irradiated groups up to a dose of 2 Gy, while a further increase in the irradiation dose resulted in a significant decline in MPCE and MNCE frequencies in both groups. Pretreatment of mice with 2mg/kg body weight of NIN resulted in a significant decline in the frequencies of MPCE and MNCE. NIN treatment not only reduced the frequency of MPCE with one micronucleus, but also of MPCE with multiple micronuclei (MN), indicating its ability to reduce complex chromosome aberrations. Conversely, the PCE/NCE ratio declined in a dose-dependent manner in both groups. The treatment of mice with NIN before exposure to different doses of gamma-radiation resulted in the inhibition in this decline in the PCE/NCE ratio. Our study demonstrates that NIN is able to protect mouse bone marrow cells against the radiation-induced DNA damage and decline in the cell proliferation as observed by a reduction in the micronucleus frequency and an increase in PCE/NCE ratio, respectively, in the NIN-pretreated irradiated group.

The accumulation of intracytoplasmic inclusion bodies (Lewy bodies) composed of aggregates of the alpha-synuclein (á-syn) protein is the principal pathological characteristic of Parkinson's disease (PD) and may lead to degeneration of dopaminergic neurons. To date there is no medication that can promote the efficient clearance of these pathological aggregates. In this study, the effect on á-syn aggregate clearance of ginkgolic acid (GA), a natural compound extracted from Ginkgo biloba leaves that inhibits SUMOylation amongst other pathways, was assessed in SH-SY5Y neuroblastoma cells and rat primary cortical neurons. Depolarization of SH-SY5Y neuroblastoma cells and rat primary cortical neurons with KCl was used to induce á-syn aggregate formation. Cells pre-treated with either GA or the related compound, anacardic acid, revealed a significant decrease in intracytoplasmic aggregates immunopositive for á-syn and SUMO-1. An increased frequency of autophagosomes was also detected with both compounds. GA post-treatment 24 h after depolarization also significantly diminished á-syn aggregate bearing cells, indicating the clearance of pre-formed aggregates. Autophagy inhibitors blocked GA-dependent clearance of á-syn aggregates, but not increased autophagosome frequency. Western analysis revealed that the reduction in á-syn aggregate frequency obtained with GA pre-treatment was accompanied by little change in the abundance of SUMO conjugates. The current findings show that GA can promote autophagy-dependent clearance of á-syn aggregates and may have potential in disease modifying therapy.

The simultaneous effects of several variables on the secondary sex ratio have been examined using data from over 3.7 million births which occurred in Japan during 1975--6. A weak and negative association between sex ratio and birth order was observed but it was not significant in the statistical sense. A negative effect of paternal age--birth order interaction was obtained when maternal age was controlled. The quadratic model is much more powerful than the linear model in explaining the sex ratio variability.

BACKGROUND: The JAK-STAT pathway is an important signaling pathway downstream of multiple cytokine and growth factor receptors. Dysregulated JAK-STAT signaling has been implicated in the pathogenesis of multiple human malignancies.OBJECTIVE: Given this pivotal role of JAK-STAT dysregulation, it is important to identify patients with an overactive JAK-STAT pathway for possible treatment with JAK inhibitors.METHODS: We developed a gene signature assay to detect overactive JAK-STAT signaling. The cancer cell line encyclopedia and associated gene-expression data were used to correlate the activation status of STAT5 with the induction of a set of STAT5 target genes.RESULTS: Four target genes were identified (PIM1, CISH, SOCS2, and ID1), the expression of which correlated significantly with pSTAT5 status in 40 hematologic tumor cell lines. In pSTAT5-positive models, the expression of the gene signature genes decreased following ruxolitinib treatment, which corresponded to pSTAT5 downmodulation. In pSTAT5-negative cell lines, neither pSTAT5 modulation nor a change in signature gene expression was observed following ruxolitinib treatment.CONCLUSIONS: The gene signature can potentially be used to stratify or enrich for patient populations with activated JAK-STAT5 signaling that might benefit from treatments targeting JAK-STAT signaling. Furthermore, the 4-gene signature is a predictor of the pharmacodynamic effects of ruxolitinib.

OBJECTIVE: Alpha-1,3-mannosyltransferase (ALG3) is an oncoprotein associated with multiple malignancies. We aimed to investigate the role and potential mechanisms of ALG3 in non-small cell lung cancer (NSCLC).METHODS: We detected the expressions of ALG3 in NSCLC tissues and adjacent tissues by RT-PCR, western blot and immunohistochemistry, respectively. Chi-square test was used to analyze the correlation between ALG3 expression and pathological paremeters. Then we used shRNA to construct a low expression model of ALG3 in NCI-H292 and NCI-H460. CCK-8 assay and transwell assay were then performed to monitor the proliferation, migration and invasion of NSCLC cells. Western blot was to detect the expression of EMT-related indicators. Further, the interaction of miR-98-5p with ALG3 was verified by luciferase reporter assay.RESULTS: The expression of ALG3 in NSCLC tissues was higher than that in normal tissues, and the increase in ALG3 expression was significantly associated with higher T stage, lymph node metastasis, and poor tissue differentiation. Patients with high ALG3 expression had a worse prognosis. ALG3 knockdown inhibited the proliferation, migration and invasion of NSCLC cells. In addition, the knockdown of ALG3 resulted in increased expression of EMT-related protein E-cadherin, while N-cadherin and Vimentin expression was decreased. Dual luciferase assay confirmed that miR-98-5p can specifically bind to the 3'UTR of ALG3 and reduces its expression and activity.CONCLUSION: ALG3 can promote the progression of NSCLC and is negatively regulated by miR-98-5p.

Despite improvements in the prognosis of osteosarcoma patients, chemotherapy fails in a considerable number of cases due to drug resistance. The development of novel agents may enhance chemosensitivity. This study explored the anticancer function of polydatin and its ability-in combination with paclitaxel-to overcome drug resistance in human osteosarcoma U-2OS and MG-63 cell lines. A cell proliferation assay (celll counting kit-8), a cell-cycle assay, an apoptosis assay (annexin V-fluorescein isothiocyanate/propidium iodide), and a cell migration assay (Transwell) were used to analyze cell activity. Western blot analysis and quantitative reverse-transcription polymerase chain reaction were performed to examine function-related mRNA and protein levels. Treatment with polydatin suppressed cell growth and migration in both cell lines. Moreover, polydatin induced cell apoptosis in both parental and paclitaxel-resistant cells, and cell-cycle arrest in the S phase. Furthermore, it altered the expression of various proteins associated with cell growth (Ki67, p21, cyclin A, cyclin E, and cyclin-dependent kinase 2), migration (matrix metalloproteinase-2 [MMP-2], MMP-9, and tissue inhibitor of metalloproteinase-1), apoptosis (poly[ADP-ribose] polymerase 1 and caspase 3), and drug resistance (p-glycoprotein 1, lung resistance-related protein 1, growth arrest and DNA damage-45á, glutathione S-transferase ð, and heat shock protein 27) in paclitaxel-resistant osteosarcoma cells. Cells transfected with myr-Akt caused obvious upregulation and activation of Akt, which were significantly attenuated via polydatin treatment. In conclusion, polydatin may enhance the chemosensitivity of osteosarcoma cells to paclitaxel.

Endotoxin is a cell wall component from Gram-negative bacteria, and inhaled endotoxin contributes significantly to the induction of airway inflammation and dysfunction. Background levels of endotoxin have not yet been extensively described. In this study, airborne endotoxin was measured with a standardized protocol in 5 types of background environment (169 samples) in Denmark from October to May. Endotoxin levels in a greenhouse (median = 13.2 EU/m3) were significantly higher than in the other environments. The air from biofuel plants (median = 5.3 EU/m3), the air on congested streets (median = 4.4 EU/m3) and on an agricultural field (median = 2.9 EU/m3) had higher endotoxin contents than the air in industrial areas (median = 1.3 EU/m3) or in towns (median = 0.33 EU/m3). Levels in industrial areas were significantly higher than in towns. A literature study revealed background levels of endotoxin on different continents between 0.063-410 EU/m3, with median or mean values between 0.063-3.6 EU/m3. Endotoxin concentrations in towns and industrial areas were higher in April and May than in autumn and winter, and were higher in October than in winter. These data of exposure in background environments and of seasonal variation are helpful for public health practitioners, epidemiologists and industrial hygienists.

We present the use of an error correcting autoencoder stage to a convolutional neural network model as a means of improving image based automatic Positive Airway Pressure (PAP) mask sizing accuracy. A single convolutional layer neural network was pre-trained using MUCT dataset and transfer learning was applied to mitigate against the relatively small custom dataset. The base model was then augmented with an additional error correcting autoencoder and trained against the custom dataset. The presented model increased PAP sizing accuracy against the baseline by 15.3% while reducing overfitting.

Knowing the proportions of asymmetric (excitatory) and symmetric (inhibitory) synapses in the neuropil is critical for understanding the design of cortical circuits. We used focused ion beam milling and scanning electron microscopy (FIB/SEM) to obtain stacks of serial sections from the six layers of the juvenile rat (postnatal day 14) somatosensory cortex (hindlimb representation). We segmented in three-dimensions 6184 synaptic junctions and determined whether they were established on dendritic spines or dendritic shafts. Of all these synapses, 87-94% were asymmetric and 6-13% were symmetric. Asymmetric synapses were preferentially located on dendritic spines in all layers (80-91%) while symmetric synapses were mainly located on dendritic shafts (62-86%). Furthermore, we found that less than 6% of the dendritic spines establish more than one synapse. The vast majority of axospinous synapses were established on the spine head. Synapses on the spine neck were scarce, although they were more common when the dendritic spine established multiple synapses. This study provides a new large quantitative dataset that may contribute not only to the knowledge of the ultrastructure of the cortex, but also towards defining the connectivity patterns through all cortical layers.

In order to investigate changes in FHR when a fetus suffers acidemia, we produced progressively advanced acidemia in lamb fetus by intermittently repeated cord compression. FHR was monitored throughout the study. FHR patterns were classified into five characteristic types as fetal arterial pH fell from around 7.35 to below 6.90. It was confirmed by studies involving catecholamine release and the administration of drugs such as atropine sulfate, alpha or beta adrenergic antagonists that sympathetic and parasympathetic regulation was deeply involved in the changes. Among the patterns, type 4, in which decreased amplitude of the initial drop and hypoxia-induced deceleration was followed by overshoot acceleration, took place during acidemia at arterial pH below 7.15. In conclusion, the appearance of type 4 FHR indicates a deteriorating state in human fetus as well, induced by repeated cord compression, and obliges us to deliver the fetus as soon as possible.

PURPOSE: To determine the feasibility of diffusion-tensor imaging (DTI) and tractography as means of evaluating the depth of mural invasion by gastric carcinomas.MATERIALS AND METHODS: This study was approved by our institutional review board, and written informed consent was obtained from each patient. Twenty gastric specimens containing a carcinoma were studied with a 7.0-T MR imaging system equipped with a four-channel phased-array surface coil. DTI was performed by using a field of view of 50-60 mm ? 25-30 mm, matrix of 256 ? 128, section thickness of 1mm, b value of 1000 s/mm2, and motion-probing gradients in seven noncollinear directions. The MR images were compared with the histopathologic findings as the reference standard.RESULTS: In all 20 carcinomas (100%) the diffusion-weighted images, apparent diffusion coefficient (ADC) maps, fractional anisotropy (FA) maps, ë1 maps, and direction-encoded color FA maps made it possible to identify the same depth of tumor invasion of the gastric wall as observed during histopathologic examination. The ë1 maps provided the best contrast between the carcinomas and the layers of the gastric wall. The carcinomas also had lower ADC values and lower FA values than the normal gastric wall; thus, the carcinomas were clearly demarcated from the normal gastric wall. Tractography images were also useful for determining the depth of tumor invasion of the gastric wall.CONCLUSIONS: DTI and tractography are feasible means of evaluating gastric specimens and provide excellent diagnostic accuracy for evaluating mural invasion by gastric carcinomas.

Soybeans are a rich source of protein and a key feed ingredient in livestock production, but lack sufficient levels of cysteine and methionine to meet the nutritional demands of swine or poultry as feed components. Although engineering the sulfur assimilatory pathway could lead to increased sulfur-containing amino acid content, little is known about this pathway in legumes. Here, we describe the cloning and characterization of soybean ATP sulfurylase (ATPS), which acts as the metabolic entry point into the sulfur assimilation pathway. Analysis of the ATPS clone isolated from a soybean seedling cDNA library revealed an open-reading frame, encoding a 52 kDa polypeptide with an N-terminal chloroplast/plastid transit peptide, which was related to the enzymes from Arabidopsis, potato, human, and yeast. Soybean ATP sulfurylase was expressed in Escherichia coli and purified to apparent homogeneity. Based on gel-filtration chromatography, the enzyme functions as a 100 kDa homodimer. Analysis of genomic DNA by Southern blotting revealed that multiple genes encode ATP sulfurylase in soybean. Analysis of the transcript profiles retrieved from a soybean EST database indicated that ATP sulfurylase mRNA was most abundant in root tissue. Cold treatment induced mRNA accumulation and enhanced the specific activity of ATP sulfurylase in root tissue. Northern blot analysis indicated a decline in the ATP sulfurylase transcript levels during seed development. Likewise, ATP sulfurylase specific activity also declined in the later stages of seed development. Increasing the expression levels of this key enzyme during soybean seed development could lead to an increase in the availability of sulfur amino acids, thereby enhancing the nutritional value of the crop.

The daughterless (da) gene in Drosophila encodes a broadly expressed transcriptional regulator whose specific functions in the control of sex determination and neurogenesis have been extensively examined. We describe here a third major developmental role for this regulatory gene: follicle formation during oogenesis. A survey of da RNA and protein distribution during oogenesis reveals a multiphasic expression pattern that includes both germline and soma. Whereas the germline expression reflects da's role in progeny sex determination, the somatic ovary expression of da correlates with the gene's role during egg chamber morphogenesis. Severe, but viable, hypomorphic da mutant genotypes exhibit dramatic defects during oogenesis, including aberrantly defined follicles and loss of interfollicular stalks. The follicular defects observed in da mutant ovaries are qualitatively very similar to those described in Notch (N) or Delta (Dl) mutant ovaries. Moreover, in the ovary da- alleles exhibit dominant synergistic interactions with N or Dl mutations. We propose that all three of these genes function in the same regulatory pathway to control follicle formation.

The hypotheses so far advanced for B-cell activation by antigen are based on the assumption that the interaction between antigen and the Ig receptors delivers at least the initial triggering signal. There are few, if any, experimental findings to support this assumption. On the contrary, a variety of findings indicate that the Ig receptors do not deliver any signal to the cells, whereas activation can be regularly achieved without participation ofIg receptors. The available evidence forces us to suggest that antigen-induced B-cell triggering is always caused by one nonspecific signal, which is delivered to the cells by surface structures, which are not the Ig receptors. For distinctive features of this hypothesis see One Nonspecific Triggering Signal. Various competing hypotheses for B-cell activation have been analyzed, but none of them appears to satisfy the experimental findings.

OBJECTIVES: Histidine-tryptophan-ketoglutarate (HTK-Custodiol) cardioplegic solution is administered as one single dose for more than 2 hours of ischemia. No prospective randomized clinical study has compared the effects of HTK and cold blood cardioplegia on myocardial damage in elective mitral valve surgery. Thus, the main aim of the present study was to examine whether one single dose of cold antegrade HTK gives as good myocardial protection as repetitive antegrade cold blood cardioplegia in mitral valve surgery.METHODS: Eighty consecutive patients undergoing elective isolated mitral valve surgery for mitral regurgitation, with or without ablation for atrial fibrillation, were included in the study and randomized to HTK or blood cardioplegia. Markers of myocardial injury (troponin-T and creatine kinase MB) were analyzed at baseline and 7 hours, 1 day, 2 days, and 3 days after surgery.RESULTS: No significant difference in creatine kinase MB and troponin-T between HTK and blood cardioplegia groups was found at any time point. There was a significant correlation between ischemic time and markers of myocardial injury in the HTK group only and significantly more spontaneous ventricular fibrillation after release of crossclamping in the HTK group.CONCLUSIONS: One single dose of antegrade cold HTK cardioplegic solution in elective mitral valve surgery protects the myocardium equally well as repetitive antegrade cold blood cardioplegia.

To study the changes in carnitine during intense muscular effort subjects underwent 4 min intermittent electrical stimulation of the quadriceps femoris muscle and on a separate occasion performed 4 min exercise on a bicycle ergometer. Biopsies of the vastus lateralis muscle were taken at rest and after 2 and 4 min of stimulation or exercise. Resting mean muscle total carnitine content was 20.0 mmol/kg dry muscle. Approximately 77% was free carnitine and 19% acetylcarnitine. Four minutes of stimulation or intense exercise did not effect total carnitine but did result in a marked fall in free carnitine and almost equivalent rise in acetylcarnitine. The results indicate that acetylcarnitine is a major metabolite formed during intense muscular effort and that carnitine may function in the regulation of the acetyl-CoA/CoA ratio by buffering excess production of acetyl units.

Unconjugated bilirubin (UCB) is responsible for neonatal jaundice and high level of free bilirubin (Bf) can lead to kernicterus. Previous studies suggest that oxidative stress is a critical component of UCB-induced neurotoxicity. The Nrf2 pathway is a powerful sensor for cellular redox state and is activated directly by oxidative stress and/or indirectly by stress response protein kinases. Activated Nrf2 translocates to nucleus, binds to Antioxidant Response Element (ARE), and enhances the up-regulation of cytoprotective genes that mediate cell survival. The aim of the present study was to investigate the role of Nrf2 pathway in cell response to bilirubin mediated oxidative stress in the neuroblastoma SH-SY5Y cell line. Cells exposed to a toxic concentration of UCB (140 nM Bf) showed an increased intracellular ROS levels and enhanced nuclear accumulation of Nrf2 protein. UCB stimulated transcriptional induction of ARE-GFP reporter gene and induced mRNA expression of multiple antioxidant response genes as: xCT, Gly1, ãGCL-m, ãGCL-c, HO-1, NQO1, FTH, ME1, and ATF3. Nrf2 siRNA decreased UCB induced mRNA expression of HO1 (75%), NQO1 (54%), and FTH (40%). The Nrf2-related HO-1 induction was reduced to 60% in cells pre-treated with antioxidant (NAC) or specific signaling pathway inhibitors for PKC, P38á and MEK1/2 (80, 40 and 25%, respectively). In conclusion, we demonstrated that SH-SY5Y cells undergo an adaptive response against UCB-mediated oxidative stress by activation of multiple antioxidant response, in part through Nrf2 pathway.

This study aimed to assess the prevalence of medical and nonmedical use of psychiatric medication among undergraduate students of health sciences from a public university in Brasil. Another objective was to determine the frequency of nuclear morphological abnormalities in the buccal mucosa of students using psychiatric drugs. A cross-sectional study based on a Web survey was carried out with 375 health sciences undergraduate students from schools of Pharmacy, Physical Education, Nutrition, and Medicine. Additionally, spontaneous genetic damages in exfoliated cells of the buccal mucosa of 41 individuals by counting micronucleus (MN) and binucleated (BN) cells frequencies were evaluated. The results showed 76 (20.3%) of students reported the use of psychotropic drugs after enrolling in university. The majority of these students were from Pharmacy and Medicine programs, females, aged between 18-25 years old, nonsmokers, alcohol addicts, and with a family history of mental illness. In addition, Medical students, individuals with high-income, who live alone and are in the last period of the program are more likely to use psychotropic drugs. Moreover, exposure to psychiatric medication was able to increase the number of binucleated cells. These results provide evidence that the use of psychoactive drugs is increased in the academic context and may be related to the failure of the cell cycle.

Realgar-Indigo naturalis formula(RIF)is an oral form of arsenic developed for treatment of acute promyelocytic leukemia (APL). We retrospectively evaluated the efficacy and safety of a novel RIF combined with all-trans retinoic acid (ATRA) based chemotherapy-free approach in newly diagnosed APL patients. Patients received oral ATRA (25 mg/m2/day in 2 divided doses) plus intravenous arsenic trioxide (0.15 mg/kg/day) or oral RIF (60 mg/kg/day in 3 divided doses) as induction chemotherapy, followed by 2 consolidations with ATRA plus RIF and maintenance therapy with intermittent ATRA and RIF. From January 2015 to December 2017, 40 subjects were enrolled. Eighteen subjects were male. Median age was 42 years (range, 14-77 years) and 10 subjects were ? 60 years. All subjects achieved a complete morphologic remission after initial induction. Molecular complete remission achieved 100% after second RIF plus ATRA consolidation. Median follow-up of survivors was 27 months (range, 7-43 months). The 2-year event-free survival (EFS) and overall survival (OS) were both 100%. Adverse events were modest and all patients needed only outpatient care during postremission therapy. Compared to our historical RIF plus ATRA with chemotherapy regimen (the Chinese APL07 trial), the inpatient treatment duration was greatly reduced by the RIF plus ATRA regimen. Our data indicates that early switching to RIF plus ATRA based chemotherapy-free approach has yielded encouraging outcomes and might be considered a practicable option to treat patients with newly diagnosed APL.

Immunodeficient mice have been used predominantly in biomedical research. Realizing that large animal species may have an enhanced ability to predict clinical outcome relative to mice, we worked to develop immunodeficient rabbits by CRISPR/Cas9. We first demonstrated that multiplex embryo transfer efficiently produced multiple lines of single-gene mutant (SGM) founders. Embryos microinjected with single sgRNA targeting FOXN1, RAG2, IL2RG or PRKDC were pooled for embryo transfer. As few as three recipients were used to produce twenty SGM founders for four genes. We then demonstrated the powerful multiplex targeting capacity of CRISPR/Cas9. First, two genes on the same chromosome were targeted simultaneously, resulting in three RAG1/RAG2 double-gene mutant (DGM) founders. Next we microinjected forty-five embryos each with five sgRNAs targeting FOXN1, RAG1, RAG2, IL2RG and PRKDC, and transferred them to two recipients. Five founders were produced: one SGM, two DGM, one triple-gene mutant and one quadruple-gene mutant. The present work demonstrates that multiplex embryo transfer and multiplex gene targeting can be used to quickly and efficiently generate mutant rabbit founders. Four lines of SGM (e.g. FOXN1, RAG2, IL2RG, and PRKDC) immunodeficient rabbits, as well as multigenic mutant immunodeficient rabbits have been produced. These animals may prove useful for biomedical research.

The lymphatic drainage of diseased and normal bowel was studied in 21 patients undergoing surgery for Crohn's disease. Mesenteric lymphatic obstruction was a consistent feature, identified in areas of small bowel macroscopically affected by Crohn's disease. This finding was also observed in some areas of apparently unaffected small bowel. Subsequent examination of these areas confirmed the presence of early Crohn's disease. This method of study may prove to be of value for determining the extent of operative resection in patients with Crohn's disease. Experimental lymphatic obstruction in animals failed to produce pathological changes of Crohn's disease, suggesting that this feature is an epiphenomenon and is not of primary importance in the pathogenesis of regional enteritis.

Interpretation of dense single nucleotide polymorphism (SNP) follow-up of genome-wide association or linkage scan signals can be facilitated by establishing expectation for the behaviour of primary mapping signals upon fine-mapping, under both null and alternative hypotheses. We examined the inferences that can be made regarding the posterior probability of a real genetic effect and considered different disease-mapping strategies and prior probabilities of association. We investigated the impact of the extent of linkage disequilibrium between the disease SNP and the primary analysis signal and the extent to which the disease gene can be physically localised under these scenarios. We found that large increases in significance (>2 orders of magnitude) appear in the exclusive domain of genuine genetic effects, especially in the follow-up of genome-wide association scans or consensus regions from multiple linkage scans. Fine-mapping significant association signals that reside directly under linkage peaks yield little improvement in an already high posterior probability of a real effect. Following fine-mapping, those signals that increase in significance also demonstrate improved localisation. We found local linkage disequiliptium patterns around the primary analysis signal(s) and tagging efficacy of typed markers to play an important role in determining a suitable interval for fine-mapping. Our findings help inform the interpretation and design of dense SNP-mapping follow-up studies, thus facilitating discrimination between a genuine genetic effect and chance fluctuation (false positive).

Phylogenetic analysis was performed for different genome regions of Japanese encephalitis virus (JEV). Similar genetic groupings were identified for all analyzed genome regions including complete genomes. More extensive analysis was performed for 92 isolates (complete envelope sequences) available in the GenBank. Results of phylogenetic analysis were compared with those performed for human positive strand RNA viruses with well characterized serotypes - poliovirus (PV) and dengue virus (DEN). The observed level of the JEV inter-genotype diversity was much less than that observed across PV and DEN serotypes and was consistent with the genetic diversity observed within PV or DEN serotypes. This genetic analysis supports the contention that all known JEV isolates comprise a single serotype.

Between 1962 and 1986, 70 patients were treated with radiation for confirmed or suspected intracranial germinoma at our hospital. The diagnosis was based on histology in 30 cases, cerebrospinal fluid (CSF) cytology in 12 cases, and on clinical and radiological findings in the remaining 28 cases. The target of radiation was the primary tumor site in 34 cases (Group A), the entire neuraxis in 22 cases (Group B), the whole brain in 4 cases (Group C), and the ventricle plus spine in 6 cases (Group D). Four patients were not included in the above groups for various reasons. The average radiation dose was 50-55 Gy to the tumor, 30 Gy to the whole brain, and 24 Gy to the spinal axis. The 5- and 10-year survival rates of the 68 primary cases in which radiotherapy was completed were 86% and 79%, respectively. The survival and relapse-free survival rates for the above 4 groups did not differ significantly, although slightly better results were seen in Groups B and C. Five cases in Groups A and D developed intracranial recurrence, 4 adjacent to the primary site but 1 distant from it, whereas no intracranial recurrence was found in the whole-brain-treated groups (B and C). One patient in Group B developed spinal metastasis, which was possibly due to inadequate radiation fields, and another in Group B developed abdominal metastasis via the shunt tube. Craniospinal irradiation should be administered to the patients with demonstrated meningeal seeding or with a positive CSF cytology. For cytology-negative cases with no evident metastasis, irradiation of the tumor plus a wide margin is usually sufficient, but craniospinal irradiation should be considered when the disease extends along the ventricular walls or is present in both pineal and suprasellar regions.

A photoactivated, site-selective conjugation of poly(ethylene glycol) (PEG) to the glutathione (GSH) binding pocket of glutathione S-transferase (GST) is described. To achieve this, a GSH analogue (GSH-BP) was designed and chemically synthesized with three functionalities: (1) the binding affinity of GSH to GST, (2) a free thiol for polymer functionalization, and (3) a photoreactive benzophenone (BP) component. Different molecular weights (2 kDa, 5 kDa, and 20 kDa) of GSH-BP modified PEGs (GSBP-PEGs) were synthesized and showed conjugation efficiencies between 52% and 76% to GST. Diazirine (DA) PEG were also prepared but gave conjugation yields lower than for GSBP-PEGs. PEGs with different end-groups were also synthesized to validate the importance of each component in the end-group design. End-groups included glutathione (GS-PEG) and benzophenone (BP-PEG). Results showed that both GSH and BP were crucial for successful conjugation to GST. In addition, conjugations of 5 kDa GSBP-PEG to different proteins were investigated, including bovine serum albumin (BSA), lysozyme (Lyz), ubiquitin (Ubq), and GST-fused ubiquitin (GST-Ubq) to ensure specific binding to GST. By combining noncovalent and covalent interactions, we have developed a new phototriggered protein-polymer conjugation method that is generally applicable to GST-fusion proteins.

Protein degradation by the ubiquitin-proteasome system is critical for neuronal function. Neurons utilize microtubule-dependent molecular motors to allocate proteasomes to synapses, but how proteasomes are coupled to motors and how this is regulated to meet changing demand for protein breakdown remain largely unknown. We show that the conserved proteasome-binding protein PI31 serves as an adaptor to couple proteasomes with dynein light chain proteins (DYNLL1/2). The inactivation of PI31 inhibited proteasome motility in axons and disrupted synaptic proteostasis, structure, and function. Moreover, phosphorylation of PI31 by p38 MAPK enhanced binding to DYNLL1/2 and promoted the directional movement of proteasomes in axons, suggesting a mechanism to regulate loading of proteasomes onto motors. Inactivation of PI31 in mouse neurons attenuated proteasome movement in axons, indicating this process is conserved. Because mutations affecting PI31 activity are associated with human neurodegenerative diseases, impairment of PI31-mediated axonal transport of proteasomes may contribute to these disorders.

BACKGROUND: To elucidate the clinical characteristics of IgG4-related dacryoadenitis.METHODS: Clinical features, laboratory findings, radiological findings, associated diseases, treatment, and prognosis were prospectively examined in 12 patients (seven men, five women; mean age, 60.9 ± 15.1 years) with IgG4-related dacryoadenitis.RESULTS: In addition to eyelid swelling, other ophthalmologic symptoms were observed in seven patients, including diplopia (n = 4), ptosis (n = 2), visual field disturbance (n = 2), eye pain (n = 2), decrease of visual acuity (n = 2), eye-movement disturbance (n = 1), dry eye (n = 1), corneal ulcer (n = 1), and epiphora (n = 1). Swelling of the lacrimal glands was bilateral in half of the patients. Other IgG4-related diseases were present in nine patients, including sialadenitis (n = 5), autoimmune pancreatitis (n = 4), retroperitoneal fibrosis (n = 2), and lymphadenopathy (n = 8). Serum IgG4 levels were significantly higher in patients with other IgG4-related disease (1070 ± 813 mg/dl) than in those without (197 ± 59 mg/dl, p = 0.017). Allergic histories and elevated serum IgE levels were each detected in six patients. Eight patients showed inflammatory extension beyond the lacrimal gland, such as thickened rectus muscle (n = 6), inflammation of the optic nerve (n = 2), and retrobulbar inflammation (n = 3). Steroid therapy was effective in seven patients, but dacryoadenitis relapsed in two patients with markedly higher serum IgG4 levels and autoimmune pancreatitis.CONCLUSIONS: IgG4-related dacryoadenitis showed various ophthalmologic symptoms due to extensive inflammation beyond the lacrimal gland, frequent association with other IgG4-related disease or allergic phenomena, and steroid responsiveness.

An alkaline phosphatase which binds to hyaluronate with a high affinity has been extracted from the bovine periodontal ligament and purified. The procedure consisted of extraction with a 10 mM Tris/Mg buffer containing 0.1% Nonidet P40, chromatography on a hyaluronate-conjugated Affi-Gel 15, and fast protein liquid chromatography. The hyaluronate-binding alkaline phosphatase has an apparent molecular weight of 110,000, contained sialic acid and was more stable to heat treatment than were two other species (Mr = 120,000 and 130,000) extracted from PDL. The heat stability and influence of inhibitors on its activity show that the hyaluronate-binding alkaline phosphatase of periodontal ligament was similar to the bone alkaline phosphatase.