Computational design of new Peptide inhibitors for amyloid Beta (aβ) aggregation in Alzheimer's disease: application of a novel methodology.
Alzheimer's disease is the most common form of dementia. It is a neurodegenerative and incurable disease that is associated with the tight packing of amyloid fibrils. This packing is facilitated by the compatibility of the ridges and grooves on the amyloid surface. The GxMxG motif is the major factor creating the compatibility between two amyloid surfaces, making it an important target for the design of amyloid aggregation inhibitors. In this study, a peptide, experimentally proven to bind Aβ40 fibrils at the GxMxG motif, was mutated by a novel methodology that systematically replaces amino acids with residues that share similar chemical characteristics and subsequently assesses the energetic favorability of these mutations by docking. Successive mutations are combined and reassessed via docking to a desired level of refinement. This methodology is both fast and efficient in providing potential inhibitors. Its efficiency lies in the fact that it does not perform all possible combinations of mutations, therefore decreasing the computational time drastically. The binding free energies of the experimentally studied reference peptide and its three top scoring derivatives were evaluated as a final assessment/valuation. The potential of mean forces (PMFs) were calculated by applying the Jarzynski's equality to results of steered molecular dynamics simulations. For all of the top scoring derivatives, the PMFs showed higher binding free energies than the reference peptide substantiating the usage of the introduced methodology to drug design.


Nuciferine ameliorates hepatic steatosis in high-fat diet/streptozocin-induced diabetic mice through a PPARα/PPARγ coactivator-1α pathway.
Nuciferine, an alkaloid found in Nelumbo nucifera leaves, alleviates dyslipidemia in vivo. However, whether it improves liver injury in diabetic conditions and the underlying mechanism is unclear. The present study aimed to investigate the effects of nuciferine on lipid and glucose metabolism in a murine model of Type 2 diabetes mellitus (T2DM) and to determine the underlying mechanisms of these effects. A murine model of T2DM was induced by high-fat diet (HFD) feeding combined with streptozocin (STZ) injections, and the diabetic mice were treated with nuciferine in their food. The underlying mechanism of the anti-steatotic effect of nuciferine was further explored in HepG2 hepatocytes cultured with palmitic acid. Major signalling profiles involved in fatty acid oxidation were then evaluated, using Western blot, RT-qPCR and si-RNA techniques, along with immunohistochemistry. Nuciferine restored impaired glucose tolerance and insulin resistance in diabetic mice. Hepatic levels of total cholesterol, triglycerides and LDL were decreased, as were the number of lipid droplets, by nuciferine treatment. Furthermore, nuciferine up-regulated β-oxidation related genes in livers of diabetic mice. Luciferase reporter cell assay showed that nuciferine directly reversed palmitic acid-induced inhibition of PPARα transcriptional activity. Silencing PPARγ coactivator-1α (PGC1α) expression in HepG2 cells abolished the effects of nuciferine in accelerating β-oxidation. Nuciferine improved lipid profile and attenuated hepatic steatosis in HFD/STZ-induced diabetic mice by activating the PPARα/PGC1α pathway. Nuciferine may be a potentially important candidate in improving hepatic steatosis and the management of T2DM.


Controversies in testosterone supplementation therapy.
Testosterone has now become one of the most widely used medications throughout the world. The rapid growth of the testosterone market in the past 10 years is due to many factors. We currently have a worldwide aging population. In the US, the number of men 65 years old or older is increasing 2-3 times faster than the number of men younger than 65 years. In addition, poor general health and certain medical conditions such as diabetes/metabolic syndrome (MetS), cardiovascular disease (CVD), and osteoporosis have been associated with low serum testosterone levels. [1],[2],[3] There are now fewer concerns regarding the development of prostate cancer (PCa) after testosterone therapy, making it a more attractive treatment option. Finally, the introduction of different forms of testosterone supplementation therapy (TST) with increased promotion, marketing, and direct-to-consumer advertising is also driving market growth. As the demand for TST continues to grow, it is becoming more important for clinicians to understand how to diagnose and treat patients with low testosterone.


[Heterogeneity of peptidoglycans of bacterial origin].
Heterogeneity of peptidoglycans of Staphylococcus aureus of strains 3272 and Wood-46, Brevibacterium flavum, Micrococcus lysodeikticus has been studied by the method of electrophoresis in the system polyacrylamide gel--sodium dodecylsulfate. It is shown that peptidoglycans of different strains of S. aureus and other species of gram-positive bacteria treated with ultrasound are a complex of fractions with 3 basic high-molecular components with M 100, 92 and 84 kDa, while two main fractions with Mm 92 and 84 kDa are identified in peptidoglycans drugs treated with ultrasound.


Modular synthesis of α-aryl β-perfluoroalkyl ketones via N-heterocyclic carbene catalysis.
A new general de novo synthesis of pharmaceutically important α-aryl β-perfluoroalkyl ketones has been disclosed. Compared with trifluoromethylation-initiated radical 1,2-aryl migration of α,α-diaryl allylic alcohols, this protocol employs a new strategy of biomimetic carbene catalysis to assemble alkene, aldehyde and perfluoroalkyl reagents, providing access to products with excellent flexibility of the aryl unit and perfluoroalkyl group. This method also demonstrates excellent functional group compatibility, including some Grignard reagent sensitive groups.


A randomised, double-blind, placebo-controlled trial of Bifidobacterium bifidum CCFM16 for manipulation of the gut microbiota and relief from chronic constipation.
A variety of opinions exist on the potential of probiotics to provide relief from chronic constipation with much focus being placed on their mechanism of action and causes of heterogeneity in the results of different studies. We aimed to determine the efficacy and safety of ingesting Bifidobacterium bifidum (B. bifidum) CCFM16 for 28 days to relieve constipation and to understand the mechanism of action. Using Rome IV criterion, 53 and 50 participants diagnosed with chronic constipation were included in the probiotic group and placebo group, respectively. Spontaneous bowel movements (SBMs) per week, stool consistency (Bristol Stool Form Scale [BSFS]), the proportion of SBM responders, patient assessment of constipation symptoms (PAC-SYM), and quality of life (PAC-QoL) were evaluated. The gut microbiota, short-chain fatty acids (SCFAs) and other indicators were also assessed. B. bifidum CCFM16 treatment improved the stool consistency and increased the proportion of SBM responders, but the differences in PAC-SYM and PAC-QoL were statistically insignificant between the groups. Analysis of the SCFAs and microbiome revealed that CCFM16 significantly increased the acetic acid and butyric acid concentrations and enhanced the Firmicutes/Bacteroidetes ratio. Levels of Clostridia were particularly increased and were associated with the increase in butyric acid. In addition, we found the other side of Clostridia; several taxa in the order Clostridiales were observed to prevent CCFM16 from proper functioning in the pre-treatment microbiome. In conclusion, CCFM16 can potentially and efficaciously relieve chronic constipation in Chinese adults by regulating the gut microbiota and SCFA metabolism. The two sides of Clostridia illustrate its importance in microbial therapy for constipation.


Inactivation and regulation of the aerobic C(4)-dicarboxylate transport (dctA) gene of Escherichia coli.
The gene (dctA) encoding the aerobic C(4)-dicarboxylate transporter (DctA) of Escherichia coli was previously mapped to the 79-min region of the linkage map. The nucleotide sequence of this region reveals two candidates for the dctA gene: f428 at 79.3 min and the o157a-o424-o328 (or orfQMP) operon at 79.9 min. The f428 gene encodes a homologue of the Sinorhizobium meliloti and Rhizobium leguminosarum H(+)/C(4)-dicarboxylate symporter, DctA, whereas the orfQMP operon encodes homologues of the aerobic periplasmic-binding protein- dependent C(4)-dicarboxylate transport system (DctQ, DctM, and DctP) of Rhodobacter capsulatus. To determine which, if either, of these loci specify the E. coli DctA system, the chromosomal f428 and orfM genes were inactivated by inserting Sp(r) or Ap(r) cassettes, respectively. The resulting f428 mutant was unable to grow aerobically with fumarate or malate as the sole carbon source and grew poorly with succinate. Furthermore, fumarate uptake was abolished in the f428 mutant and succinate transport was approximately 10-fold lower than that of the wild type. The growth and fumarate transport deficiencies of the f428 mutant were complemented by transformation with an f428-containing plasmid. No growth defect was found for the orfM mutant. In combination, the above findings confirm that f428 corresponds to the dctA gene and indicate that the orfQMP products play no role in C(4)-dicarboxylate transport. Regulation studies with a dctA-lacZ (f428-lacZ) transcriptional fusion showed that dctA is subject to cyclic AMP receptor protein (CRP)-dependent catabolite repression and ArcA-mediated anaerobic repression and is weakly induced by the DcuS-DcuR system in response to C(4)-dicarboxylates and citrate. Interestingly, in a dctA mutant, expression of dctA is constitutive with respect to C(4)-dicarboxylate induction, suggesting that DctA regulates its own synthesis. Northern blot analysis revealed a single, monocistronic dctA transcript and confirmed that dctA is subject to regulation by catabolite repression and CRP. Reverse transcriptase-mediated primer extension indicated a single transcriptional start site centered 81 bp downstream of a strongly predicted CRP-binding site.


The brain adjusts grip forces differently according to gravity and inertia: a parabolic flight experiment.
In everyday life, one of the most frequent activities involves accelerating and decelerating an object held in precision grip. In many contexts, humans scale and synchronize their grip force (GF), normal to the finger/object contact, in anticipation of the expected tangential load force (LF), resulting from the combination of the gravitational and the inertial forces. In many contexts, GF and LF are linearly coupled. A few studies have examined how we adjust the parameters-gain and offset-of this linear relationship. However, the question remains open as to how the brain adjusts GF regardless of whether LF is generated by different combinations of weight and inertia. Here, we designed conditions to generate equivalent magnitudes of LF by independently varying mass and movement frequency. In a control experiment, we directly manipulated gravity in parabolic flights, while other factors remained constant. We show with a simple computational approach that, to adjust GF, the brain is sensitive to how LFs are produced at the fingertips. This provides clear evidence that the analysis of the origin of LF is performed centrally, and not only at the periphery.


Telehealth policy: looking for global complementarity.
Telehealth is gaining acceptance as a tool for bridging the local and global health-care divides. However, integrating telehealth into existing health infrastructures presents a daunting challenge for governments, policy makers, telehealth advocates and health-care workers. The development of specific inter-jurisdictional telehealth policies will significantly improve the ability to meet this challenge. In the policy context, one 'success' is the increasing number of jurisdictions addressing policy issues. However, policy decisions have largely been taken in isolation, within individual health institutions, regions, provinces/states or countries. This represents a failure of the current approach. Telehealth, by its very nature, has the ability to transgress existing geo-political boundaries. As a consequence, policy in any single jurisdiction may hamper or even cripple the ability of telehealth to fulfil its potential. Commonality--or at least complementarity--of approach to telehealth policy must be encouraged. To achieve this, it is essential to understand the current or anticipated regulatory constraints that may affect telehealth. We have begun a preliminary study of country-specific policy issues.


Hispanic race is a risk factor for COVID-19 during pregnancy: data from an urban New York City hospital.
There are limited studies on predisposing factors for COVID-19 positivity in asymptomatic pregnant women. The literature published to date on asymptomatic COVID-19 pregnant carriers does not focus on pregnancy or pre-pregnancy comorbidities. We wanted to identify risk factors for COVID-19 in asymptomatic pregnant women. We performed a retrospective chart review of 263 asymptomatic pregnant women admitted to labour and delivery at New York City Health + Hospitals/Lincoln.We analysed the association between race, body mass index (BMI), smoking, indication for admission, gravidity, parity, pre-pregnancy comorbidity, pregnancy comorbidity via uni- and multivariate statistical tests. Only Hispanic race was significant in the univariate analysis (p = .049). At the post-hoc analysis, Hispanics had a higher proportion of COVID-19 cases compared to non-Hispanic Blacks (p = .019). No variables were significantly associated with COVID-19 positivity in the multivariate analysis.Hispanic race appears to be a risk factor for asymptomatic COVID-19 infection during pregnancy. We speculate that the cultural and socioeconomic reality of Hispanic women living in our community leads to more exposure opportunities and therefore, a higher infection rate.Impact statementWhat is already known on this subject? Little is known on the role of comorbidities and risk factors that can favour COVID-19 infection during pregnancy.What do the results of this study add? We found that Hispanic pregnant asymptomatic women had a higher rate of COVID-19 in comparison to non-Hispanic Black women. Pre-pregnancy comorbidities such as pregestational diabetes, hypertension and asthma were not associated with COVID-19 positivity.What are the implications of these findings for clinical practice and/or further research? The reasons why the Hispanic race is more affected by COVID-19 during pregnancy is unclear. The social environment of Hispanic women living in our community, such as their tendency to live in multigenerational and multi-family households, might contribute to a higher infection rate. More resources might be dedicated in the future to Hispanic-dense neighbourhoods.


Klebsiella pneumoniae carbapenemase variants: the new threat to global public health.
SUMMARYKlebsiella pneumoniae carbapenemase (KPC) variants, which refer to the substitution, insertion, or deletion of amino acid sequence compared to wild blaKPC type, have reduced utility of ceftazidime-avibactam (CZA), a pioneer antimicrobial agent in treating carbapenem-resistant Enterobacterales infections. So far, more than 150 blaKPC variants have been reported worldwide, and most of the new variants were discovered in the past 3 years, which calls for public alarm. The KPC variant protein enhances the affinity to ceftazidime and weakens the affinity to avibactam by changing the KPC structure, thereby mediating bacterial resistance to CZA. At present, there are still no guidelines or expert consensus to make recommendations for the diagnosis and treatment of infections caused by KPC variants. In addition, meropenem-vaborbactam, imipenem-relebactam, and other new β-lactam-β-lactamase inhibitor combinations have little discussion on KPC variants. This review aims to discuss the clinical characteristics, risk factors, epidemiological characteristics, antimicrobial susceptibility profiles, methods for detecting blaKPC variants, treatment options, and future perspectives of blaKPC variants worldwide to alert this new great public health threat.


Effect of electrical field strength applied by PEF processing and storage temperature on the outgrowth of yeasts and moulds naturally present in a fresh fruit smoothie.
Pulsed electrical field (PEF) technology offers an alternative to thermal pasteurisation of high-acid fruit juices, by extending the shelf life of food products, while retaining its fresh taste and nutritional value. Substantial research has been performed on the effect of electrical field strength on the inactivation kinetics of spoilage and pathogenic micro-organisms and on the outgrowth of spoilage micro-organisms during shelf life. However, studies on the effect of electrical field strength on the inactivation and outgrowth of surviving populations during shelf life are missing. In this study, we assessed the influence of electrical field strength applied by PEF processing and storage temperature on the outgrowth of surviving yeast and mould populations naturally present in fresh fruit smoothie in time. Therefore, an apple-strawberry-banana smoothie was treated in a continuous-flow PEF system (130L/h), using similar inlet and outlet conditions (preheating temperature 41°C, maximum temperature 58°C) to assure that the amount of energy across the different conditions was kept constant. Smoothies treated with variable electrical field strengths (13.5, 17.0, 20.0 and 24.0kV/cm) were compared to smoothies without treatment for outgrowth of yeasts and moulds. Outgrowth of yeasts and moulds stored at 4°C and 7°C was analysed by plating and visual observation and yeast growth was modelled using the modified logistic growth model (Zwietering model). Results showed that the intensity of the electrical field strength had an influence on the degree of inactivation of yeast cells, resulting in a faster outgrowth over time at lower electrical field strength. Outgrowth of moulds over time was not affected by the intensity of the electrical field strength used. Application of PEF introduces a trade-off between type of spoilage: in untreated smoothie yeasts lead to spoilage after 8days when stored at 4 or 7°C, whereas in PEF treated smoothie yeasts were (partly) inactivated and provided outgrowth opportunities for moulds, which led to spoilage by moulds after 14days (7°C) or 18days (4°C).


Incident cataract after a five-year interval and lifestyle factors: the Beaver Dam eye study.
A population-based study to investigate risk factors for age-related eye disease was begun in 1987 in a representative American community. Incidence of cataract was subsequently evaluated. To examine the relationships of cigarette smoking, alcohol, and caffeine intakes to incidence of age-related cataracts five years later. Observational epidemiologic incidence study of an adult population. Adults 43-84 years of age were identified during a census in 1987-1988, and examined at baseline (1988-1990) and after a five-year interval (1993-1995). Standardized protocols were used at the baseline and follow-up evaluations for exposures and for objective identification of cataracts. Cumulative incidence of nuclear cataract in right eyes was about 12%, cortical cataract about 8%, and posterior subcapsular cataract about 3%. In multivariable analyses of the general estimating equations type, there were significant associations between nuclear cataract and: pack-years smoked (odds ratio [OR] 1.05, 95% confidence interval [CI] 1.01, 1.09 per 10 pack-years); and current alcohol intake (OR 1.01, 95% CI 1.00, 1.02 per 10 grams ethanol/week). Cigarette smokers were more likely to have had cataract surgery in the interval between baseline and follow-up examinations. Cigarette smoking and alcohol consumption were associated with modestly increased risks of incident nuclear cataract over a five-year interval.


Normal variations in the lateral contour of the head and neck of the pancreas mimicking neoplasm: evaluation with dual-phase helical CT.
The purpose of this study was to analyze variations in the lateral contour of the head and neck of the pancreas that can mimic pancreatic masses on CT imaging. We retrospectively reviewed dual-phase helical CT examinations of 119 patients who had no clinical or CT evidence of pancreatic disease. Contour variations of the head and neck of the pancreas were analyzed and were classified according to their anatomic orientation. Forty-one (34.5%) of the 119 patients had discrete lobulations of pancreatic tissue greater than 1 cm lateral to the gastroduodenal or anterior superior pancreaticoduodenal artery. These lobulations showed normal pancreatic density on both predominantly arterial and portal venous phase images. Contour variants of the pancreatic head and neck were categorized as three main types: anterior (type I), posterior (type II), and horizontal (type III). In the 119 patients, we found 12 type I variants (10%), 23 type II variants (19%), and six type III variants (5%). Variations in the lateral contour of the normal head and neck of the pancreas are common. Recognition of the different types of contours may help avoid misinterpretation of normal variants as pancreatic masses.


Stigma, Expressed Emotion, and Quality of Life in Caregivers of Individuals with Dementia.
Expressed emotion (EE) is a measure of a caregiver's critical and emotionally overinvolved (EOI; e.g., intrusive, self-sacrificing) attitudes and behaviors toward a person with a mental illness. Mounting evidence indicates that high levels of these critical and EOI attitudes and behaviors (collectively termed high EE) in family members are associated with a poorer course of illness for people with a range of disorders, including dementia (Nomura et al., 2005). However, less is known about factors that might trigger high EE and how high EE might impact dementia caregivers' own mental health. In this study we propose that caregivers who perceive stigma from their relative's illness may be more likely to be critical or intrusive (high EOI) toward their relative in an attempt to control symptomatic behaviors. We further hypothesized that high EE would partially mediate the link between stigma and quality of life (QoL) as there is some evidence that high EE is associated with poorer mental health in caregivers themselves (Safavi et al., 2015). In line with study hypotheses and using a sample of 106 dementia caregivers, we found that greater caregiver stigma was associated with both high EE (for criticism and EOI) and with poorer QoL. Mediational analyses further confirmed that high EE accounts for much of the association between stigma and poorer QoL. Study results suggest that addressing caregiver stigma in therapy could reduce levels of high EE and indirectly therefore improve caregiver QoL. Intervening directly to reduce high EE could also improve caregiver QoL.


Interactions within and between clades shaped the diversification of terrestrial carnivores.
A longstanding debate in evolutionary biology and paleontology is whether ecological interactions such as competition impose diversity dependence on speciation and extinction rates. Here, we analyze the fossil record of terrestrial mammalian carnivores in North America and Eurasia using a Bayesian framework to assess whether their diversity dynamics were affected by diversity dependence within and between families (12 in Eurasia, 10 in North America). We found eight instances of within-clade diversity dependence suppressing speciation rates and detected between-clade effects increasing extinction rates in six instances. Diversity dependence often involved lineages that migrated between continents and we found that speciation was more responsive to diversity changes within the clade, whereas extinction responded to diversity of taxa in other clades. The analysis of the fossil record of Carnivora suggests that interactions within and between clades are associated with different speciation and extinction regimes, opening room for a broader theory of diversity dependence.


Effects of ethanol extraction and heat treatment of soybean flakes on function and morphology of pig intestine.
Digesta and tissue samples were collected from the intestinal tracts of 27 pigs to determine the relationship between intestinal morphology and the utilization of nutrients from soybean flakes. Soybean flake treatments were under-, intermediate- and over-processed (i.e., 5, 20 and 60 min of autoclaving) either without extraction or with heating before or after extraction with a 55% ethanol-water mixture. Final BW was greatest (P less than .001) for pigs fed soybean flakes given 20 min of heat treatment. There was a trend (P less than .09) for plasma lysine concentrations to be reduced when the unextracted soybean flakes were over-processed (60 min of heat). Differences in the flow rate of DM and N through the ileum and colon reflected differences in DM and N intake, rather than differences in intestinal function. The soybean flake treatments had no effect (P greater than .08) on pH of the contents of the stomach, duodenum, ileum or colon. The ethanol extraction process increased (P less than .001) N digestibility of the soybean flakes, especially when the soybean flakes were underprocessed (interaction, P less than .02). Villus size (area, height and perimeter length) tended to be greater in pigs fed the soybean flakes heated after extraction and(or) exposed to the intermediate level of heat treatment. Indicators of villus shape (villus area/villus height) and proliferative activity (crypt depth and villus height/crypt depth) were not affected by soybean flake treatment (P greater than .08). Ethanol extraction and heat treatment affected the utilization of nutrients from soybean flakes.(ABSTRACT TRUNCATED AT 250 WORDS)


Gene regulatory systems that control gene expression in the Ciona embryo.
Transcriptional control of gene expression is one of the most important regulatory systems in animal development. Specific gene expression is basically determined by combinatorial regulation mediated by multiple sequence-specific transcription factors. The decoding of animal genomes has provided an opportunity for us to systematically examine gene regulatory networks consisting of successive layers of control of gene expression. It remains to be determined to what extent combinatorial regulation encoded in gene regulatory networks can explain spatial and temporal gene-expression patterns. The ascidian Ciona intestinalis is one of the animals in which the gene regulatory network has been most extensively studied. In this species, most specific gene expression patterns in the embryo can be explained by combinations of upstream regulatory genes encoding transcription factors and signaling molecules. Systematic scrutiny of gene expression patterns and regulatory interactions at the cellular resolution have revealed incomplete parts of the network elucidated so far, and have identified novel regulatory genes and novel regulatory mechanisms.


Estimating successive pKa values of polyprotic acids from ab initio molecular dynamics using metadynamics: the dissociation of phthalic acid and its isomers.
Estimation of the dissociation constant, or pKa, of weak acids continues to be a central goal in theoretical chemistry. Here we show that ab initio Car-Parrinello molecular dynamics simulations in conjunction with metadynamics calculations of the free energy profile of the dissociation reaction can provide reasonable estimates of the successive pKa values of polyprotic acids. We use the distance-dependent coordination number of the protons bound to the hydroxyl oxygen of the carboxylic group as the collective variable to explore the free energy profile of the dissociation process. Water molecules, sufficient to complete three hydration shells surrounding the acid molecule, were included explicitly in the computation procedure. Two distinct minima corresponding to the dissociated and un-dissociated states of the acid are observed and the difference in their free energy values provides the estimate for pKa, the acid dissociation constant. We show that the method predicts the pKa value of benzoic acid in good agreement with experiment and then show using phthalic acid (benzene dicarboxylic acid) as a test system that both the first and second pKa values as well, as the subtle difference in their values for different isomers can be predicted in reasonable agreement with experimental data.


Esophageal candidiasis in HIV-negative patients.
Candida esophagitis is the most frequent esophageal infection in HIV seropositive as well as in seronegative patients. this retrospective study was designed to determine the characteristics of this disease in HIV negative patients in a general hospital. clinical records of all HIV negative patients with Candida esophagitis, which was endoscopically diagnosed and microscopically confirmed (biopsy and/or cytology of esophageal mucosa), were studied. thirty-one patients (23 men, 8 women, mean age: 65.4 +/- 14.3 years, median 71) fulfilled the criteria (0.56% of the diagnostic esophagogastroduodenal endoscopies). The most common clinical symptoms were dysphagia with or without odynophagia and pain (retrosternal, epigastric or xiphoid). Fourteen patients (45%) had no esophageal signs; in nine of them (29%) the disease was diagnosed in the course of an endoscopic exam to investigate the sources of acute or chronic anaemia. The most frequent predisposing factors were diabetes mellitus, oral or aerosolized corticotherapy, malignancies, treatment with broad-spectrum antibiotics and liver cirrhosis. Nine patients (29%) presented no known predisposing factors. The most common endoscopic appearance was grade II (51%). Sixty-one per cent of the patients exhibited at least one other esophagogastroduodenal endoscopic lesion associated with the mycosis. Three patients died of their underlying diseases within one week of the diagnosis of the candidiasis. Only one patient presented persistence of candidiasis one week after initiating treatment with oral nystatine. esophageal candidiasis is infrequent and does not always present with suspicious symptoms or known predisposing factors. Aerosolized corticotherapy may be a risk factor for the development of esophageal candidiasis.


[Unified National Health System costs in São José dos Campos, São Paulo State, Brazil, for hospital admissions due to external causes].
The objective of this study was to identify direct expenditures for hospitalizations due to external causes in the Unified National Health System (SUS) in the city of São José dos Campos, São Paulo State, Brazil. Admissions to the Dr. José de Carvalho Florence Municipal Hospital resulting from external causes or injuries - ICD-10, chapters XIX and XX respectively - were analyzed for the first semester of 2003. 976 patient admission forms were analyzed, after data evaluation. Admissions with the highest total cost were those resulting from motor vehicle accidents and falls. The highest mean cost for hospitalization for injury was due to motor vehicle accidents (BRL 614.63), followed by assault (BRL 594.90). The highest mean cost for hospitalization due to injury was for cervical fractures (BRL 1,191.42) and head injuries (BRL 1,000.44). Hospitalizations with the highest daily cost were skull and facial fractures (BRL 166.72) and abdominal trauma (BRL 148.26). The study confirmed that motor vehicle accidents, falls, and assault are an important source of costs due to hospitalization for injuries.


Why do African elephants (Loxodonta africana) simulate oestrus? An analysis of longitudinal data.
Female African elephants signal oestrus via chemicals in their urine, but they also exhibit characteristic changes to their posture, gait and behaviour when sexually receptive. Free-ranging females visually signal receptivity by holding their heads and tails high, walking with an exaggerated gait, and displaying increased tactile behaviour towards males. Parous females occasionally exhibit these visual signals at times when they are thought not to be cycling and without attracting interest from musth males. Using demographic and behavioural records spanning a continuous 28-year period, we investigated the occurrence of this "simulated" oestrus behaviour. We show that parous females in the Amboseli elephant population do simulate receptive oestrus behaviours, and this false oestrus occurs disproportionately in the presence of naïve female kin who are observed coming into oestrus for the first time. We compare several alternative hypotheses for the occurrence of this simulation: 1) false oestrus has no functional purpose (e.g., it merely results from abnormal hormonal changes); 2) false oestrus increases the reproductive success of the simulating female, by inducing sexual receptivity; and 3) false oestrus increases the inclusive fitness of the simulating female, either by increasing the access of related females to suitable males, or by encouraging appropriate oestrus behaviours from female relatives who are not responding correctly to males. Although the observed data do not fully conform to the predictions of any of these hypotheses, we rule out the first two, and tentatively suggest that parous females most likely exhibit false oestrus behaviours in order to demonstrate to naïve relatives at whom to direct their behaviour.


[Standardized testing of bone implant surfaces with an osteoblast cell culture cyste. III. PVD hard coatings and Ti6Al4V].
The effect of titanium-based PVD coatings and a titanium alloy on the proliferation and differentiation of osteoblasts was investigated using a standardised cell culture system. Human fetal osteoblasts (hFOB 1.19) were cultured on titanium-niobium-nitride ([Ti,Nb]N), titanium-niobium-oxy-nitride coatings ([Ti,Nb]ON) and titanium-aluminium-vanadium alloy (Ti6Al4V) for 17 days. Cell culture polystyrene (PS) was used as reference. For the assessment of proliferation, the numbers and viability of the cells were determined, while alkaline phosphatase activity, collagen I and osteocalcin synthesis served as differentiation parameters. On the basis of the cell culture experiments, a cytotoxic effect of the materials can be excluded. In comparison with the other test surfaces, [Ti,Nb]N showed greater cell proliferation. The [Ti,Nb]N coating was associated with the highest level of osteocalcin production, while all other differentiation parameters were identical on all three surfaces. The test system described reveals the influence of PVD coatings on the osteoblast differentiation cycle. The higher oxygen content of the [Ti,Nb]ON surface does not appear to have any positive impact on cell proliferation. The excellent biocompatibility of the PVD coatings is confirmed by in vivo findings. The possible use of these materials in the fields of osteosynthesis and articular surfaces is still under discussion.


The Early Language in Victoria Study: predicting vocabulary at age one and two years from gesture and object use.
The Macarthur-Bates Communicative Development Inventories (CDI) have been used widely to document early communicative development. The paper reports on a large community sample of 1,447 children recruited from low, middle and high socioeconomic (SES) areas across metropolitan Melbourne, Australia. Regression analyses were conducted to determine the extent to which communicative behaviours reported at 0 ; 8 and 1 ; 0 predicted vocabulary development at 1 ; 0 and 2 ; 0. In support of previous findings with smaller, often less representative samples, gesture and object use at 1 ; 0 were better predictors of 2 ; 0 vocabulary than were gesture and object use at 0 ; 8. At 1 ; 0, children from the lower SES groups were reported to understand more words than children from the higher SES groups, but there were no SES differences for words produced at 1 ; 0 or 2 ; 0. The findings add to our understanding of the variability in the development of early communicative behaviours.


Meiotic silencing and fragmentation of the male germline restricted chromosome in zebra finch.
During male meiotic prophase in mammals, X and Y are in a largely unsynapsed configuration, which is thought to trigger meiotic sex chromosome inactivation (MSCI). In avian species, females are ZW, and males ZZ. Although Z and W in chicken oocytes show complete, largely heterologous synapsis, they too undergo MSCI, albeit only transiently. The W chromosome is already inactive in early meiotic prophase, and inactive chromatin marks may spread on to the Z upon synapsis. Mammalian MSCI is considered as a specialised form of the general meiotic silencing mechanism, named meiotic silencing of unsynapsed chromatin (MSUC). Herein, we studied the avian form of MSUC, by analysing the behaviour of the peculiar germline restricted chromosome (GRC) that is present as a single copy in zebra finch spermatocytes. In the female germline, this chromosome is present in two copies, which normally synapse and recombine. In contrast, during male meiosis, the single GRC is always eliminated. We found that the GRC in the male germline is silenced from early leptotene onwards, similar to the W chromosome in avian oocytes. The GRC remains largely unsynapsed throughout meiotic prophase I, although patches of SYCP1 staining indicate that part of the GRC may self-synapse. In addition, the GRC is largely devoid of meiotic double strand breaks. We observed a lack of the inner centromere protein INCENP on the GRC and elimination of the GRC following metaphase I. Subsequently, the GRC forms a micronucleus in which the DNA is fragmented. We conclude that in contrast to MSUC in mammals, meiotic silencing of this single chromosome in the avian germline occurs prior to, and independent of DNA double strand breaks and chromosome pairing, hence we have named this phenomenon meiotic silencing prior to synapsis (MSPS).


Preconception venous thromboembolism and placenta-mediated pregnancy complications.
Placenta-mediated complications are leading causes of maternal and fetal morbidity and mortality. We hypothesized that a preconception history of venous thromboembolism (VTE) is associated with increased risk of placenta-mediated pregnancy complications. A nationwide population-based cohort study of all singleton pregnancies leading to delivery from 1997 to 2012 (n = 964 967). We obtained data on placenta-mediated pregnancy complications from the Danish Medical Birth Registry and data on VTE before pregnancy from the Danish National Patient Registry. We computed absolute risks, crude and adjusted risk differences (RDs) using a binomial regression model, and crude and adjusted risk ratios (RRs) from a modified Poisson regression model. Overall, 1419 women had a preconception history of VTE, while 578 112 did not. Preeclampsia occurred in 4.2% of pregnancies in the VTE group and in 2.7% of pregnancies in a comparison cohort (adjusted RD = 1.3%, 95% confidence interval (CI) 0.6-2.0%; adjusted RR = 1.5, 95% CI 1.3-1.8). Stillbirth occurred in 0.7% of pregnancies in the VTE group and in 0.4% of pregnancies in the comparison cohort (adjusted RD = 0.3%, 95% CI 0.02-0.6%; adjusted RR = 1.8, 95% CI 1.1-3.0). Placental abruption occurred in 0.8% of pregnancies in the VTE group and in 0.5% of pregnancies in the comparison cohort (adjusted RD = 0.3%, 95% CI - 0.05-0.6%; adjusted RR = 1.6, 95% CI 1.1-2.4). Small-for-gestational-age infants accounted for 10.9% of live births in the VTE group and 9.8% of live births in the comparison cohort (adjusted RD = 0.6%, 95% CI - 0.5-1.7%; adjusted RR = 1.1, 95% CI 0.9-1.3). Women with a history of VTE were at increased risk of placenta-mediated complications.


TNFRSF11A-Associated Dysosteosclerosis: A Report of the Second Case and Characterization of the Phenotypic Spectrum.
Dysosteosclerosis (DOS) is a distinct form of sclerosing bone disease characterized by irregular osteosclerosis and platyspondyly. DOS is genetically heterogeneous; however, only five cases with SLC29A3 mutations and a single case with a splice-site mutation of TNFRSF11A have been reported, and TNFRSF11A is also a causal gene for osteopetrosis, autosomal recessive 7 (OP-AR7). Thus, the causal genes of DOS and their genotype-phenotype associations remain unclear. In this study, we examined a Japanese patient with DOS and found a novel variant in TNFRSF11A. The homozygous variant was a G to T transversion at the first nucleotide of exon 9 (c.784G>T). Although the variant was predicted to cause a stop codon mutation (p.E262*), in silico evaluation of the exonic splicing elements followed by RT-PCR for the patient-derived cells showed that it caused aberrant splicing due to the change in the exonic splicing element and produced two types of aberrant transcripts: One caused a premature stop codon (p.E262Vfs*17) leading to nonsense mutation-mediated mRNA decay; the other produced a protein with interstitial deletion (p.E262_Q279del). The effects of the mutation on five splicing isoforms of TNFRSF11A were different from those in OP-AR7, but comparable with those in the first DOS with the TNFRSF11A mutation. Thus, we identified the second case of DOS caused by the TNFRSF11A splice-site mutation and confirmed the novel disease entity. © 2019 American Society for Bone and Mineral Research.


Protective effect of probiotics on Salmonella infectivity assessed with combined in vitro gut fermentation-cellular models.
Accurate assessment of probiotics with targeted anti-Salmonella activity requires suitable models accounting for both, microbe-microbe and host-microbe interactions in gut environments. Here we report the combination of two original in vitro intestinal models closely mimicking the complex in vivo conditions of the large intestine. Effluents from continuous in vitro three-stage fermentation colonic models of Salmonella Typhimurium infection inoculated with immobilized child microbiota and Salmonella were directly applied to confluent mucus-secreting HT29-MTX cell layers. The effects of Salmonella, addition of two bacteriocinogenic strains, Bifidobacterium thermophilum RBL67 (thermophilicin B67) and Escherichia coli L1000 (microcin B17), and inulin were tested on Salmonella growth and interactions with epithelial cell layers. Salmonella adhesion and invasion were investigated and epithelial integrity assessed by transepithelial electrical resistance (TER) measurements and confocal microscopy observation. Data from complex effluents were compared with pure Salmonella cultures. Salmonella in effluents of all reactors of the colonic fermentation model stabilized at mean values of 5.3 ± 0.8 log10 cfu/ml effluent. Invasion of cell-associated Salmonella was up to 50-fold lower in complex reactor samples compared to pure Salmonella cultures. It further depended on environmental factors, with 0.2 ± 0.1% being measured with proximal, 0.6 ± 0.2% with transverse and 1.3 ± 0.7% with distal reactor effluents, accompanied by a similar high decrease of TER across cell monolayers (minus 45%) and disruption of tight junctions. Subsequent addition of E. coli L1000 stimulated Salmonella growth (6.4 ± 0.6 log10 cfu/ml effluent of all 3 reactors) and further decreased TER, but led to 10-fold decreased invasion efficiency when tested with distal reactor samples. In contrast, presence of B. thermophilum RBL67 revealed a protective effect on epithelial integrity compared to previous E. coli L1000 periods, as reflected by a significant mean increase of TER by 58% in all reactors. Inulin addition enhanced Salmonella growth and invasion when tested with distal and proximal reactor samples, respectively, but induced a limited decrease of TER (minus 18%) in all reactors. Our results highlight the benefits of combining suitable cellular and colonic fermentation models to assess strain-specific first-level host protection properties of probiotics during Salmonella infection, providing an efficient system biology tool for preclinical development of new antimicrobials.


Incidences, predictors, and clinical outcomes of acute and late stent malapposition detected by optical coherence tomography after drug-eluting stent implantation.
We investigated the incidences, predictors, and clinical outcomes of acute and late stent malapposition detected by optical coherence tomography (OCT) after drug-eluting stent implantation. We analyzed the OCT images from 351 patients with 356 lesions who received poststent and follow-up OCT examinations. Acute stent malapposition was observed in 62% of lesions. Approximately half of the acute stent malappositions were located within the edges of the stents. Severe diameter stenosis, calcified lesions, and long stents were independent predictors of acute stent malapposition. Follow-up OCT examinations were performed 175±60 days after drug-eluting stent implantation. Thirty-one percent of lesions with acute stent malapposition remained malapposed (late-persistent stent malapposition) and were typically (72%) located within the edges of the stent. The location within the stent edges and the volume of acute stent malapposition were independent predictors of late-persistent stent malapposition. Acute stent malapposition with a volume >2.56 mm(3) differentiated late-persistent stent malapposition from resolved acute stent malapposition. Late-acquired stent malapposition was detected in 15% of all lesions and was usually (61%) located within the stent body. Late-acquired stent malapposition was more frequently associated with plaque/thrombus prolapse on poststent OCT images (70% versus 42%; P<0.001). Clinical events, including cardiovascular death, nonfatal myocardial infarction, and stent thrombosis, did not occur in patients with late stent malapposition during the follow-up period of 28.6±10.3 months after drug-eluting stent implantation. Acute, late-persistent, and late-acquired stent malapposition had relatively high incidences but different predictors. The clinical outcome of stent malapposition was favorable.


Intrathecal chemoprophylaxis after HSCT in children.
At present, the literature on the efficacy and risks of i.t. chemotherapy to children after HSCT is scarce. Current practices to reduce the risk of leukemic relapse in the CNS after HSCT differ between centers of transplantation. We compared 74 patients (56 ALL/18 AML), who received i.t. therapy post-HSCT with 46 patients (36 ALL/10 AML) who did not receive post-HSCT i.t. therapy. The patients were transplanted at the University Children's Hospital, Uppsala or the Karolinska University Hospital, Huddinge, two Swedish transplantation units with different routines concerning i.t. therapy after HSCT. The primary end-point was the number of isolated CNS relapses. Secondary end-points were other types of relapse, death, and neurological complications. There was no statistically significant difference in the incidence of CNS relapses between the groups (p > 0.05). I.t. therapy did not reduce the overall incidence of isolated CNS relapse or mortality. Our study did not demonstrate a protective effect of i.t. therapy indicating that post-HSCT i.t. therapy may only be of limited use in the treatment of acute childhood leukemia. We conclude that with the risks present, i.t. therapy should be carefully evaluated, and only considered in high-risk cases.


Sol-gel polycondensation in a cyclohexane-based organogel system in helical silica: creation of both right- and left-handed silica structures by helical organogel fibers.
Chiral amide- and urea-type organic gelators (1-6), based on cyclohexanediamine, have been prepared, and the superstructures of the organogels were evaluated by circular dichroism (CD), transmission electron microscopy (TEM), and scanning electron microscopy (SEM). The CD spectrum of the amide-based organogel system 1+2 exhibited a negative sign for the first Cotton effect, indicating that the dipole moments in the gelator aggregate orient into an anticlockwise direction. In contrast, the system 3+4 has a positive sign for the first Cotton effect, indicating that they orient into a clockwise direction. In the mixture of urea- and amide-based organogels, CD spectra of 5+2 and 6+4 revealed negative and positive signs, respectively. The helical structure of the amide-based organogels 1+2 and 3+4 clearly showed left- and right-handed structures, respectively, by SEM. To transcribe the chiral, helical structures of the organogels into silica gel, the sol-gel polycondensation of tetraethoxysilane (TEOS) was carried out in acetonitrile or ethanol. Very surprisingly, the left- and right-handed structures of the silica can be created by transcription of left- and right-handed structures of the organogels 1+2 (R enantiomer) and 3+4 (S enantiomer), respectively. In addition, in the mixture systems of urea- and amide-based organogels, the right- and left-handed structures of the organogels 5+2 (R enantiomer) and 6+4 (S enantiomer) were precisely transcribed into the silica structure. The findings suggest that the sol-gel polycondensation proceeds along the surface of the helical structure of the organogels. We therefore believe that the sol-gel polycondensation by means of molecular assembly templates abundantly built in the organogel phase is a new strategy to create superstructured silica materials.


β-Actin shows limited mobility and is required only for supraphysiological insulin-stimulated glucose transport in young adult soleus muscle.
Studies in skeletal muscle cell cultures suggest that the cortical actin cytoskeleton is a major requirement for insulin-stimulated glucose transport, implicating the β-actin isoform, which in many cell types is the main actin isoform. However, it is not clear that β-actin plays such a role in mature skeletal muscle. Neither dependency of glucose transport on β-actin nor actin reorganization upon glucose transport have been tested in mature muscle. To investigate the role of β-actin in fully differentiated muscle, we performed a detailed characterization of wild type and muscle-specific β-actin knockout (KO) mice. The effects of the β-actin KO were subtle; however, we confirmed the previously reported decline in running performance of β-actin KO mice compared with wild type during repeated maximal running tests. We also found insulin-stimulated glucose transport into incubated muscles reduced in soleus but not in extensor digitorum longus muscle of young adult mice. Contraction-stimulated glucose transport trended toward the same pattern, but the glucose transport phenotype disappeared in soleus muscles from mature adult mice. No genotype-related differences were found in body composition or glucose tolerance or by indirect calorimetry measurements. To evaluate β-actin mobility in mature muscle, we electroporated green fluorescent protein (GFP)-β-actin into flexor digitorum brevis muscle fibers and measured fluorescence recovery after photobleaching. GFP-β-actin showed limited unstimulated mobility and no changes after insulin stimulation. In conclusion, β-actin is not required for glucose transport regulation in mature mouse muscle under the majority of the tested conditions. Thus, our work reveals fundamental differences in the role of the cortical β-actin cytoskeleton in mature muscle compared with cell culture.


An immersive journey to the molecular structure of SARS-CoV-2: Virtual reality in COVID-19.
The era of the explosion of immersive technologies has bumped head-on with the coronavirus disease 2019 (COVID-19) global pandemic caused by the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2). The proper understanding of the three-dimensional structures that compose the virus, as well as of those involved in the infection process and in treatments, is expected to contribute to the advance of fundamental and applied research against this pandemic, including basic molecular biology studies and drug design. Virtual reality (VR) is a powerful technology to visualize the biomolecular structures that are currently being identified for SARS-CoV-2 infection, opening possibilities to significant advances in the understanding of the disease-associate mechanisms and thus to boost new therapies and treatments. The present availability of VR for a large variety of practical applications together with the increasingly easiness, quality and economic access of this technology is transforming the way we interact with digital information. Here, we review the software implementations currently available for VR visualization of SARS-CoV-2 molecular structures, covering a range of virtual environments: CAVEs, desktop software, and cell phone applications, all of them combined with head-mounted devices like cardboards, Oculus Rift or the HTC Vive. We aim to impulse and facilitate the use of these emerging technologies in research against COVID-19 trying to increase the knowledge and thus minimizing risks before placing huge amounts of money for the development of potential treatments.


Emergence of motor synergy in vertical reaching task via tacit learning.
The dynamics of multijoint limbs often causes complex dynamic interaction torques which are the inertial effect of other joints motion. It is known that Cerebellum takes important role in a motor learning by developing the internal model. In this paper, we propose a novel computational control paradigm in vertical reaching task which involves the management of interaction torques and gravitational effect. The obtained results demonstrate that the proposed method is valid for acquiring motor synergy in the system with actuation redundancy and resulted in the energy efficient solutions. It is highlighted that the tacit learning in vertical reaching task can bring computational adaptability and optimality with model-free and cost-function-free approach differently from previous studies.


[Biomarkers and PET for assessing the activity of Takayasu's arteritis].
Takayasu's arteritis is a chronic granulomatous vasculitis affecting the medium and large blood vessels that can lead to vascular occlusion. Inflammatory stages occur during the course of the disease. Angiography remains the gold standard examination of choice but less invasive techniques are needed to assess the activity of the disease and to monitor response to treatment. We present a case of an asymptomatic 21 year-old female patient with occlusion and stenosis of supra-aortic trunks discovered while performing an ultrasonography for the study of a recent goiter. Angiography confirmed stenosis and vascular occlusions, Takayasu's arteritis being diagnosed. Metalloprotease 9 levels at diagnosis were higher than in control patients and increased when symptoms began. Positron emission tomography performed after starting treatment showed an increase in uptake. These non-invasive markers could be useful to assess the activity of the disease and even to avoid invasive angiographic examinations.


Characterization of the Tyrosine Kinase-Regulated Proteome in Breast Cancer by Combined use of RNA interference (RNAi) and Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) Quantitative Proteomics.
Tyrosine kinases (TKs) are central regulators in cellular activities and perturbations of TK signaling contribute to oncogenesis. However, less than half of the TKs have been thoroughly studied and a global functional analysis of their proteomic portrait is lacking. Here we conducted a combined approach of RNA interference (RNAi) and stable isotope labeling with amino acids in cell culture (SILAC)-based quantitative proteomics to decode the TK-regulated proteome and associated signaling dynamics. As a result, a broad proteomic repertoire modulated by TKs was revealed, upon silencing of the 65 TKs expressed in MCF7 breast cancer cells. This yielded 10 new distinctive TK clusters according to similarity in TK-regulated proteome, each characterized by a unique signaling signature in contrast to previous classifications. We provide functional analyses and identify critical pathways for each cluster based on their common downstream targets. Analysis of different breast cancer subtypes showed distinct correlations of each cluster with clinical outcome. From the significantly up- and down-regulated proteins, we identified a number of markers of drug sensitivity and resistance. These data supports the role of TKs in regulating major aspects of cellular activity, but also reveals redundancy in signaling, explaining why kinase inhibitors alone often fail to achieve their clinical aims. The TK-SILACepedia provides a comprehensive resource for studying the global function of TKs in cancer.


Biomechanical response of the human foot when standing in a natural position while exposed to vertical vibration from 10-200 Hz.
Exposure to foot-transmitted vibration (FTV) can lead to pain and numbness in the toes and feet, increased cold sensitivity, blanching in the toes, and joint pain. Prolonged exposure can result in a clinical diagnosis of vibration-induced white foot (VIWFt). Data on the biomechanical response of the feet to FTV is limited; therefore, this study seeks to identify resonant frequencies for different anatomical locations on the human foot, while standing in a natural position. A laser Doppler vibrometer was used to measure vertical (z-axis) vibration on 21 participants at 24 anatomical locations on the right foot during exposure to a sine sweep from 10-200 Hz with a peak vertical velocity of 30 mm/s. The most notable differences in the average peak frequency occur between the toes (range: 99-147 Hz), midfoot (range: 51-84 Hz) and ankle (range: 16-39 Hz). Practitioner Summary: The biomechanical response of the human foot exposed to foot-transmitted vibration, when standing in a natural position, was measured for 21 participants. The foot does not respond uniformly; the toes, midfoot, and ankle regions need to be considered independently in future development of isolation strategies and protective measures.


A randomised controlled trial of cognitive behaviour therapy in adolescents with major depression treated by selective serotonin reuptake inhibitors. The ADAPT trial.
To determine if, in the short term, depressed adolescents attending routine NHS Child and Adolescent Mental Health Services (CAMHS), and receiving ongoing active clinical care, treatment with selective serotonin reuptake inhibitors (SSRIs) plus cognitive behaviour therapy (CBT) compared with SSRI alone, results in better healthcare outcomes. A pragmatic randomised controlled trial (RCT) was conducted on depressed adolescents attending CAMHS who had not responded to a psychosocial brief initial intervention (BII) prior to randomisation. Six English CAMHS participated in the study. A total of 208 patients aged between 11 and 17 years were recruited and randomised. All participants received active routine clinical care in a CAMHS outpatient setting and an SSRI and half were offered CBT. The duration of the trial was a 12-week treatment phase, followed by a 16-week maintenance phase. Follow-up assessments were at 6, 12 and 28 weeks. The primary outcome measure was the Health of the Nation Outcome Scales for Children and Adolescents (HoNOSCA). Secondary outcome measures were self-report depressive symptoms, interviewer-rated depressive signs and symptoms, interviewer-rated psychosocial impairment and clinical global impression of response to treatment. Information on resource use was collected in interview at baseline and at the 12- and 28-week follow-up assessments using the Child and Adolescent Service Use Schedule (CA-SUS). Of the 208 patients randomised, 200 (96%) completed the trial to the primary end-point at 12 weeks. By the 28-week follow-up, 174 (84%) participants were re-evaluated. Overall, 193 (93%) participants had been assessed at one or more time points. Clinical characteristics indicated that the trial was conducted on a severely depressed group. There was significant recovery at all time points in both arms. The findings demonstrated no difference in treatment effectiveness for SSRI + CBT over SSRI only for the primary or secondary outcome measures at any time point. This lack of difference held when baseline and treatment characteristics where taken into account (age, sex, severity, co-morbid characteristics, quality and quantity of CBT treatment, number of clinic attendances). The SSRI + CBT group was somewhat more expensive over the 28 weeks than the SSRI-only group (p=0.057) and no more cost-effective. Over the trial period there was on average a decrease in suicidal thoughts and self-harm compared with levels recorded at baseline. There was no significant increase in disinhibition, irritability and violence compared with levels at baseline. Around 20% (n=40) of patients in the trial were non-responders. Of these, 17 (43%) showed no improvement by 28 weeks and 23 (57%) were considered minimally (n=10) or moderately to severely worse (n=13). For moderately to severely depressed adolescents who are non-responsive to a BII, the addition of CBT to fluoxetine plus routine clinical care does not improve outcome or confer protective effects against adverse events and is not cost-effective. SSRIs (mostly fluoxetine) are not likely to result in harmful adverse effects. The findings are broadly consistent with existing guidelines on the treatment of moderate to severe depression. Modification is advised for those presenting with moderate (6-8 symptoms) to severe depressions (>8 symptoms) and in those with either overt suicidal risk and/or high levels of personal impairment. In such cases, the time allowed for response to psychosocial interventions should be no more than 2-4 weeks, after which fluoxetine should be prescribed. Further research should focus on evaluating the efficacy of specific psychological treatments against brief psychological intervention, determining the characteristics of patients with severe depression who are non-responsive to fluoxetine, relapse prevention in severe depression and improving tools for determining treatment responders and non-responders.


[The effect of muscle activity on the blood neutrophil system in rats].
The effect of muscular activity (MA) on the amount of different classes of leukocytes in the blood, concentrations of myeloperoxidase and lysozyme in the blood plasma, and neutrophils and phagocytosis parameters of these cells, were studied in rats. The MA resulted in a decrease of marker proteins (myeloperoxidase and lysozyme) concentration in neutrophils and an increase of their concentrations in the blood plasma. The neutrophil phagocytic activity decreased. The restoration of myeloperoxidase content in neutrophils took place within 12 hrs after MA, phagocytic activity restored by the 3rd day. The mechanisms of change of the neutrophil functional activity under the MA influence are discussed.


Utilization of IncP-1 plasmids as vectors for transposon mutagenesis in myxobacteria.
No free plasmid has ever been found in the myxobacterium Myxococcus xanthus, but IncP-1 plasmids are able to integrate into the chromosome of this bacterium. The frequency of integration depends greatly upon the structure of the IncP-1 plasmid used. This property has been used to devise new delivery systems for transposon mutagenesis in this species. Plasmids with low integration efficiencies have proved to be efficient donors of Tn5, while plasmids with very high frequencies of integration could be used directly to generate mutations. These vectors have also proved efficient for Tn5 transfer into other species of myxobacteria, which have not so far been susceptible to genetic analysis.


[Recommendations for the evaluation of complicated aphakia in private accident insurance].
An evaluation of cases in which an expert opinion was obtained during the last 2 years shows that only one-third of patients with traumatic aphakia and a 20/30 loss of vision are adequately assessed. If there are further consequences of the accident (complicated aphakia) they should also be taken into account when assessing loss of vision. A simple method of doing this is described on the basis of three examples.


[The physiological mechanisms of the neurohumoral regulation of vascular tonus].
Neuronal organisation of hypothalamo-bulbar mechanisms of regulation of vascular tonus was studied in anesthetised and immobilised cats. The descending influence of posterior, tuberal and anterior structures of the hypothalamus on the activity of antidromically identified sympatho-activating reticulospinal neurons of the ventrolateral region of medulla oblongata is realised by mono-, oligo- and polysynaptic mechanisms. Chronic neurogenic hypertension was induced in rats by overloading the higher nervous activity. Arterial hypertension did not develop in chemically desympathized cats. Central and peripheral neurohumoral mechanisms of vascular tonus regulation are discussed.


Migration-selection balance and local adaptation of mitochondrial haplotypes in rufous-collared sparrows (Zonotrichia capensis) along an elevational gradient.
Variable selection pressures across heterogeneous landscapes can lead to local adaptation of populations. The extent of local adaptation depends on the interplay between natural selection and gene flow, but the nature of this relationship is complex. Gene flow can constrain local adaptation by eroding differentiation driven by natural selection, or local adaptation can itself constrain gene flow through selection against maladapted immigrants. Here we test for evidence that natural selection constrains gene flow among populations of a widespread passerine bird (Zonotrichia capensis) that are distributed along an elevational gradient in the Peruvian Andes. Using multilocus sequences and microsatellites screened in 142 individuals collected along a series of replicate transects, we found that mitochondrial gene flow was significantly reduced along elevational transects relative to latitudinal control transects. Nuclear gene flow, however, was not similarly reduced. Clines in mitochondrial haplotype frequency were strongly associated with transitions in environmental variables along the elevational transects, but this association was not observed for the nuclear markers. These results suggest that natural selection constrains mitochondrial gene flow along elevational gradients and that the mitonuclear discrepancy may be due to local adaptation of mitochondrial haplotypes.


Molecular determinants of intrinsic resistance to doxorubicin in human cancer cell lines.
Intrinsic or acquired drug resistance poses a major challenge to the success of chemotherapy in the clinical management of human cancers. While acquired multidrug resistance (MDR), whereby cells become refractory to multiple drugs, has been extensively investigated, the mechanistic basis for intrinsic resistance remains elusive, so that this condition is largely unmanageable in the clinical setting. To address this issue, we have assessed the effects of the anticancer agent doxorubicin (DX) on a panel of human tumor cell lines originally derived from untreated patients and tried to establish a correlation between cell response and a number of parameters, including drug accumulation and/or drug efflux; differences in expression and/or subcellular distribution of proteins involved in the apoptotic process (e.g., p53, Bcl-2, Bax) and intracellular signal transducers (PKCalpha); changes in key detoxification processes. Based on our results, 'classic' multispecific drug transporters (P-glycoprotein, MDR-related proteins) only seem to play a minor role in the intrinsically resistant phenotype, whereas LRP may contribute to resistance in non-small cell lung carcinoma (NSCLC) cells. No relationship was observed between drug response and expression and/or subcellular localization of apoptosis-related proteins; however, increased PKCalpha levels are associated with poor drug response, suggesting that one or more substrates of this enzyme may be relevant to the resistant phenotype. Finally, overactive glutathione-recycling pathways may contribute to DX resistance.


Selective impairment in GLUT4 translocation to transverse tubules in skeletal muscle of streptozotocin-induced diabetic rats.
We previously reported that insulin induces the translocation of GLUT4 to both the plasma membrane and the transverse tubules (T-tubules) in rat skeletal muscle (Am J Physiol 270:E667-E676, 1996). The aim of the present study was to investigate whether the insulin-resistant glucose utilization of skeletal muscle from streptozotocin (STZ)-induced diabetic rats is linked to an impaired translocation of GLUT4 to the plasma membrane, the T-tubules, or both surface compartments. Whole-body insulin-mediated glucose disposal, assessed during a hyperinsulinemic-euglycemic clamp, was reduced by 48% (P < 0.01) in diabetic rats as compared with controls. Subcellular membrane fractions enriched with plasma membranes, T-tubules, or GLUT4-enriched intracellular membranes were isolated from hindlimb muscles of control and insulin-stimulated rats, and GLUT4 content was measured by Western blot analysis. In the absence of insulin (unstimulated), GLUT4 content in muscle of diabetic rats was markedly lower (by approximately 40%) in both the T-tubules and the intracellular membrane fraction as compared with controls. In contrast, the transporter protein levels were similar in the plasma membrane fraction. In skeletal muscle of control animals, the hyperinsulinemic clamp induced GLUT4 translocation from the intracellular membrane pool to both the plasma membrane and the T-tubule-enriched fractions (approximately 2.2-fold to approximately 2.5-fold). Surprisingly, insulin increased plasma membrane GLUT4 content to comparable levels in control and diabetic rat skeletal muscle. However, insulin-mediated GLUT4 translocation to the T-tubules was significantly reduced in the same muscle. Whole-body insulin action was significantly correlated with GLUT4 protein levels in the T-tubules, but not with the transporter content in either plasma membranes or intracellular membranes. These results strongly suggest that peripheral resistance to insulin action on glucose disposal in STZ-induced diabetic rats is caused by a selective impairment of GLUT4 translocation to skeletal muscle T-tubules.


Prevalence and Data Availability of Early Childhood Caries in 193 United Nations Countries, 2007-2017.
To assess the relationship between health care system and economic factors and early childhood caries (ECC) data availability and prevalence. We estimated ECC data for 193 United Nations countries from studies published between 2007 and 2017. We obtained other variables from the World Health Organization and the World Bank databases. We assessed association with ECC data availability by using logistic regression and with ECC prevalence by using linear regression. We included 190 publications from 88 (45.6%) countries. The mean ECC prevalence was 23.8% and 57.3% in children younger than 36 months and children aged 36 to 71 months, respectively. The odds of ECC data availability were significantly higher for countries with more physicians and more dentists. In children younger than 36 months, ECC prevalence was associated with universal health coverage (B = -6.56). In children aged 36 to 71 months, it was associated with growth of gross national income (B = 0.27). Countries with more physicians and more dentists were more likely to have ECC data. Among those with data, countries with higher economic growth had higher ECC prevalence.


[Impact of the summer 2003 heat wave on the activity of two psychiatric emergency departments].
Heat waves have been related with lethal effects, especially in Europe during the intensely hot summer of 2003. However, besides increased deaths and ailments, there are no specific data on the psychiatric effects of heat waves. We have compared psychiatric emergencies in Barcelona during a 15-day heat waves period with the rest of the 2003 summer days. The main variables of the study were total emergencies, admissions, diagnoses, Severity of Psychiatric Illness scale (SPI), psychosocial variables, treatment rendered (including use of restraints), and referrals. No differences were found in the number of emergencies and admissions. During the heat wave, there were more patients with psychiatric backgrounds, more diagnoses of alcohol and drug abuse, but fewer anxiety disorders. The proportion of patients with mechanical restraint increased, but this only occurred in half of the cases in patients with drug or alcohol abuse. The item "dangerousness toward others" (part of the SPI scale) scored significantly higher during the heat waves. There were no significant increases or decreases in psychiatric emergencies or admissions. However, the heat wave was related to more violent behavior and higher drug and alcohol abuse. It should be noted that anxiety conditions and benzodiazepine prescriptions were lower during this period. These findings may be useful to implement medical-psychiatric preventive measures against the heat wave phenomenon.


Bradyrhizobium agreste sp. nov., Bradyrhizobium glycinis sp. nov. and Bradyrhizobium diversitatis sp. nov., isolated from a biodiversity hotspot of the genus Glycine in Western Australia.
Strains of the genus Bradyrhizobium associated with agronomically important crops such as soybean (Glycine max) are increasingly studied; however, information about symbionts of wild Glycine species is scarce. Australia is a genetic centre of wild Glycine species and we performed a polyphasic analysis of three Bradyrhizobium strains-CNPSo 4010T, CNPSo 4016T, and CNPSo 4019T-trapped from Western Australian soils with Glycine clandestina, Glycine tabacina and Glycine max, respectively. The phylogenetic tree of the 16S rRNA gene clustered all strains into the Bradyrhizobium japonicum superclade; strains CNPSo 4010T and CNPSo 4016T had Bradyrhizobium yuanmingense CCBAU 10071T as the closest species, whereas strain CNPSo 4019T was closer to Bradyrhizobium liaoningense LMG 18230T. The multilocus sequence analysis (MLSA) with five housekeeping genes-dnaK, glnII, gyrB, recA and rpoB-confirmed the same clusters as the 16S rRNA phylogeny, but indicated low similarity to described species, with nucleotide identities ranging from 93.6 to 97.6% of similarity. Considering the genomes of the three strains, the average nucleotide identity and digital DNA-DNA hybridization values were lower than 94.97 and 59.80 %, respectively, with the closest species. In the nodC phylogeny, strains CNPSo 4010T and CNPSo 4019T grouped with Bradyrhizobium zhanjiangense and Bradyrhizobium ganzhouense, respectively, while strain CNPSo 4016T was positioned separately from the all symbiotic Bradyrhizobium species. Other genomic (BOX-PCR), phenotypic and symbiotic properties were evaluated and corroborated with the description of three new lineages of Bradyrhizobium. We propose the names of Bradyrhizobium agreste sp. nov. for CNPSo 4010T (=WSM 4802T=LMG 31645T) isolated from Glycine clandestina, Bradyrhizobium glycinis sp. nov. for CNPSo 4016T (=WSM 4801T=LMG 31649T) isolated from Glycine tabacina and Bradyrhizobium diversitatis sp. nov. for CNPSo 4019T (=WSM 4799T=LMG 31650T) isolated from G. max.


Astrocytes, but not olfactory ensheathing cells or Schwann cells, promote myelination of CNS axons in vitro.
We have examined the interaction between olfactory ensheathing cells (OECs), Schwann cells (SC), oligodendrocytes, and CNS axons using cultures generated from embryonic rat spinal cord. Oligodendrocyte process extension and myelination in these cultures was poor if the cells were plated on OECs or SCs. Myelin internodes and nodes of Ranvier formed frequently if these cultures were plated onto monolayers of neurosphere-derived astrocytes (NsAs). In the myelinated fibers generated on NsAs, Nav channels, caspr, and neurofascin molecules were correctly assembled at the nodes of Ranvier. The density of neurites, survival, and antigenic differentiation of oligodendrocytes was similar on OEC and NsAs monolayers. However, on OEC monolayers, despite a transient increase in the number of endogenous oligodendrocytes, there was a decrease in oligodendrocyte process extension and axonal ensheathment when compared with cultures plated on NsAs monolayers. To determine if these changes were due to axonal or glial factors, spinal cord oligodendrocytes were plated onto monolayers of OECs, NsAs, and poly-L-lysine in the absence of neurons. In these cultures, process extension and myelin-like membrane formation by oligodendrocytes was improved on monolayers of OEC. This suggests that inhibition of process extension is mediated via cross-talk between OECs and neurites. In cultures containing axons plated on OEC monolayers, oligodendrocyte process formation, axonal ensheathment, and myelination occurred albeit lower if the cultures were supplemented with NsAs conditioned medium. These data suggest OECs can permit neurite extension and oligodendrocyte proliferation, but lack secreted factor(s) and possible cell-cell contact that is necessary for oligodendrocyte process extension and myelination.


Scalar time domain modeling and coupling of second harmonic generation process in GaAs discontinuous optical waveguide.
We present in this work the scalar potential formulation of second harmonic generation process in chi((2)) nonlinear analysis. This approach is intrinsically well suited to the applications of the concept of circuit analysis and synthesis to nonlinear optical problems, and represents a novel alternative method in the analysis of nonlinear optical waveguide, by providing a good convergent numerical solution. The time domain modeling is applied to nonlinear GaAs asymmetrical waveguide with dielectric discontinuities in the hypothesis of quasi phase matching condition in order to evaluate the efficiency conversion of the second harmonic signal. The accuracy of the modeling is validated by the good agreement with the published experimental results. The effective dielectric constant method allows to extend the analysis also to 3D optical waveguides.


Preliminary Validity Evidence for a Milestones-Based Rating Scale for Chart-Stimulated Recall.
Minimally anchored Standard Rating Scales (SRSs), which are widely used in medical education, are hampered by suboptimal interrater reliability. Expert-derived frameworks, such as the Accreditation Council for Graduate Medical Education (ACGME) Milestones, may be helpful in defining level-specific anchors to use on rating scales. We examined validity evidence for a Milestones-Based Rating Scale (MBRS) for scoring chart-stimulated recall (CSR). Two 11-item scoring forms with either an MBRS or SRS were developed. Items and anchors for the MBRS were adapted from the ACGME Internal Medicine Milestones. Six CSR standardized videos were developed. Clinical faculty scored videos using either the MBRS or SRS and following a randomized crossover design. Reliability of the MBRS versus the SRS was compared using intraclass correlation. Twenty-two faculty were recruited for instrument testing. Some participants did not complete scoring, leaving a response rate of 15 faculty (7 in the MBRS group and 8 in the SRS group). A total of 529 ratings (number of items × number of scores) using SRSs and 540 using MBRSs were available. Percent agreement was higher for MBRSs for only 2 of 11 items-use of consultants (92 versus 75, P = .019) and unique characteristics of patients (96 versus 79, P = .011)-and the overall score (89 versus 82, P < .001). Interrater agreement was 0.61 for MBRSs and 0.51 for SRSs. Adding milestones to our rating form resulted in significant, but not substantial, improvement in intraclass correlation coefficient. Improvement was inconsistent across items.


Evaluation of corneal deformation analyzed with Scheimpflug based device in healthy eyes and diseased ones.
This study was designed to evaluate the correlation between corneal biomechanical and morphological data in healthy eyes, eyes that underwent myopic photorefractive keratectomy (PRK), keratoconus affected eyes, and keratoconus affected eyes that underwent corneal collagen crosslinking (CCC). Complete clinical eye examination of all eyes was followed by tomographic (Pentacam, Oculus, Wetzlar, Germany) and biomechanical (Corvis ST, Oculus, Wetzlar, Germany) evaluation. Differences among Corvis ST (CST) parameters in the different groups have been performed. Linear regression between central corneal thickness (CCT), intraocular pressure (IOP), and anterior corneal curvature measured with Sim'K (KM), versus corneal deformation parameters measured with Corvis ST in the different groups, has been run using SPSS software version 18.0. We evaluated 64 healthy eyes of 64 patients with a mean refractive error of -0.65 ± 1.68 D (measured as spherical equivalent), 17 eyes of 17 patients that underwent myopic PRK for a mean refractive defect of -4.91 ± 2.05 D (measured as spherical equivalent), 16 eyes of 16 patients affected by keratconus (stage 2-3 of Amsler Classification), and 13 eyes of 13 patients affected by keratoconus that underwent CCC. Our data suggest that corneal curvature would have a greater influence on corneal deformation than CCT; in fact KM values are more strongly associated with more CST parameters both about corneal change in shape and both about the corneal ability to come back at original shape.


Gastrointestinal phosphate handling in CKD and its association with cardiovascular disease.
Increases in serum concentrations of parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF-23) and ultimately phosphate and decreases in 1,25-dihydroxyvitamin D level are thought to play a central role in the progressive nature of kidney disease and the development of cardiovascular disease in patients with chronic kidney disease. The initial changes in PTH and FGF-23 levels are adaptive to maintain serum phosphate concentration and phosphate load within defined levels by increasing urinary excretion of phosphate. Less well appreciated is the unanticipated finding that absorption of phosphate from the gastrointestinal tract is not downregulated in chronic kidney disease. This maladaptive response maintains higher levels of phosphate absorption, thereby contributing to the phosphate burden. Moreover, in response to a low-phosphate diet, as often is prescribed to such patients, gut phosphate absorption may be enhanced, undermining the potential beneficial effects of this intervention. Given the poor response to limiting phosphate intake and the use of phosphate binders, we suggest that research efforts be oriented toward better understanding of the factors that affect phosphate absorption in the gastrointestinal tract and the development of agents that directly inhibit phosphate transporters in the small intestine and/or their associated binding proteins.


Data dictionary services in XNAT and the Human Connectome Project.
The XNAT informatics platform is an open source data management tool used by biomedical imaging researchers around the world. An important feature of XNAT is its highly extensible architecture: users of XNAT can add new data types to the system to capture the imaging and phenotypic data generated in their studies. Until recently, XNAT has had limited capacity to broadcast the meaning of these data extensions to users, other XNAT installations, and other software. We have implemented a data dictionary service for XNAT, which is currently being used on ConnectomeDB, the Human Connectome Project (HCP) public data sharing website. The data dictionary service provides a framework to define key relationships between data elements and structures across the XNAT installation. This includes not just core data representing medical imaging data or subject or patient evaluations, but also taxonomical structures, security relationships, subject groups, and research protocols. The data dictionary allows users to define metadata for data structures and their properties, such as value types (e.g., textual, integers, floats) and valid value templates, ranges, or field lists. The service provides compatibility and integration with other research data management services by enabling easy migration of XNAT data to standards-based formats such as the Resource Description Framework (RDF), JavaScript Object Notation (JSON), and Extensible Markup Language (XML). It also facilitates the conversion of XNAT's native data schema into standard neuroimaging vocabularies and structures.


Sinking, Swimming, or Learning to Swim in Medicare Part D.
Under Medicare Part D, senior citizens choose prescription drug insurance offered by numerous private insurers. We examine nonpoor enrollees' actions in 2006 and 2007 using panel data. Our sample reduced overspending by $298 on average, with gains by 81 percent of them. The greatest improvements were by those who overspent most in 2006 and by those who switched plans. Decisions to switch depended on individuals' overspending in 2006 and on individual-specific effects of changes in their current plans. The oldest consumers and those initiating medications for Alzheimer's disease improved by more than average, suggesting that real-world institutions help overcome cognitive limitations.


Analysis of carbohydrates in Lupinus albus stems on imposition of water deficit, using porous graphitic carbon liquid chromatography-electrospray ionization mass spectrometry.
This work reports the development and application of a negative ion mode online LC-ESI-MS method for studying the effect of water deficit on the carbohydrate content of Lupinus albus stems, using a porous graphitic carbon (PGC) stationary phase and an ion trap mass spectrometer. Using this method, separation and detection of several water soluble carbohydrates, ranging from mono-, di-, and oligosaccharides (raffinose, stachyose, and verbascose) to sugar alcohols was achieved in approximately 10 min. This on-line PGC-LC-ESI-MS method shows good linearity with correlation coefficients R(2)>0.99, selectivity, short analysis time, and limits of detection (LOD) ranging from 0.4 to 9 pmol for sugars and 4-20 pmol for sugar alcohols. This PGC-LC-ESI-MS method is sensitive and allowed us to detect even small alterations in carbohydrate levels in L. albus stems that resulted from a mild/early water deficit (nmol g(-1)DW). This paper describes details of our method and its application to the quantitative analysis of water soluble underivatised carbohydrates extracted from L. albus stem tissues that have been subjected to early and severe water deficit conditions, followed by a rewatering period.


Long term follow-up of patients with ventricular high rate events detected on remote monitoring of pacemakers.
Previous data suggest ventricular high rate episodes (VHREs) on pacemakers are frequent and not associated with overall mortality on short term follow up. We sought to determine whether VHREs are associated with mortality, device upgrade, or change in ejection fraction on long term follow up. A single center, retrospective study was performed on 542 patients with permanent pacemakers followed between 2011 and 2013. Follow-up was extended to 2017 for determination of long term outcomes. "True" VHREs were defined as episodes adjudicated to be due to non-sustained ventricular tachycardia on review of electrograms and "false" VHREs were defined as supraventricular arrhythmias or noise. VHRE occurred in 202(37.2%)/542 included patients. True VHRE was detected in 148(27.3%) while 54(10%) had false VHRE. The mean age of the population was 72 ± 15 years and 46% were women. Mean follow-up was 3.3 ± 1.4 years. The baseline characteristics of the true, false and no VHRE patients were similar. There was no difference in all-cause mortality between groups (27% mortality in true VHRE, 33% in false VHRE and 29% in no VHRE). Furthermore, there was no difference between groups with regards to any device upgrade (5% any upgrades in the VHRE, 9% in false VHRE and 5% in no VHRE.) On follow up, EF declined in all groups: -4% vs -2.4% vs -3.5% for true, false and no VHRE. VHRE are frequently encountered on remote monitoring of pacemakers and not associated with increased risk of mortality or need for downstream device upgrade.


Identification and characterization of novel NMDA receptor antagonists selective for NR2A- over NR2B-containing receptors.
NR1/NR2A is a subtype of N-methyl-d-aspartate receptors (NMDARs), which are glutamate and glycine-gated Ca(2+)-permeable channels highly expressed in the central nervous system. A high-throughput screening (HTS) campaign using human osteosarcoma (U-2 OS) cells transiently transduced with NR1/NR2A NMDAR subunits, tested in a specifically designed fluorometric imaging plate reader (FLIPR)/Ca(2+) assay, identified sulfonamide derivative series, exemplified by 3-chloro-4-fluoro-N-[(4-{[2-(phenylcarbonyl)hydrazino]carbonyl}phenyl)methyl]benzenesulfonamide (compound 1) and thiodiazole derivative N-(cyclohexylmethyl)-2-({5-[(phenylmethyl)amino]-1,3,4-thiadiazol-2-yl}thio)acetamide (compound 13) as novel NR1/NR2A receptor antagonists. Compounds 1 and 13 displayed submicromolar and micromolar potency at NR1/NR2A receptor, respectively, although they did not show activity at NR2B-containing receptor up to 50 μM concentration. Addition of 1 mM glycine, but not 1 mM l-glutamate, was able to surmount compound 1 and 13 inhibitory effects in FLIPR NR1/NR2A assay. However, compounds 1 and 13 displaced a glutamate site antagonist [(3)H]d,l-(E)-2-amino-4-propyl-5-phosphono-3-pentenoic acid ([(3)H]CGP 39653) to a greater extent than the glycine site antagonist [(3)H]3-[(E)-2-carboxy-2-phenylethenyl]-4,6-dichloro-1H-indole-2-carboxylic acid ([(3)H]MDL 105,519), in rat brain cortex binding assay. Results of FLIPR cell-based, electrophysiological, and biochemical binding assays suggest that compounds 1 and 13 are the prototypes of novel classes of NMDAR ligands, which to the best of our knowledge are the first selective antagonists at NR1/NR2A over NR1/NR2B receptor, and might constitute useful tools able to elucidate the relative role of the NR2A subunit in physiological and pathological conditions.


The EMT/ITK/TSK (EMT) tyrosine kinase is activated during TCR signaling: LCK is required for optimal activation of EMT.
Functional T lymphocyte activation requires concurrent stimulation of the TCR complex and an accessory molecule, most frequently CD28. We have previously demonstrated that the TEC family tyrosine kinase EMT/ITK/TSK (EMT) is activated following cross-linking of CD28. We demonstrate herein that cross-linking of the CD3 component of the TCR complex also leads to EMT activation as indicated by a rapid and transient increase in EMT tyrosine phosphorylation and kinase activity in anti-EMT immunoprecipitates. However, although concurrent cross-linking of the TCR and CD28 results in a marked increase in production of the T cell growth factor IL-2, it does not result in a significant alteration in the magnitude or duration of EMT activation. Somatic cell mutants of the Jurkat T cell line, which lack the SRC family kinase LCK (JCaM1.6), fail to produce IL-2 when stimulated through the TCR complex. EMT activation, as evidenced by increased EMT tyrosine phosphorylation and EMT-associated kinase activity, was also greatly reduced following stimulation of the TCR in the JCaM1.6 Jurkat T cell mutants that lack LCK. In support of a role for LCK in EMT activation, reconstitution of the LCK-negative Jurkat T cell line by enforced expression of LCK restored TCR-mediated EMT activation. Taken together, the data indicate that the EMT tyrosine kinase is activated following cross-linking of the TCR, a process in which LCK likely plays an important role.


Experimental Metabolic Syndrome Model Associated with Mechanical and Structural Degenerative Changes of the Aortic Valve.
The purpose of this study was to test the hypothesis that an experimental high fat (HF) animal with metabolic syndrome results in structural degeneration of the aortic valve. Domestic pigs were divided (n = 12) and administered either a normal or HF diet. After 16-weeks, the HF diet group had increased weight (p ≤ 0.05), total cholesterol (p ≤ 0.05), and systolic and diastolic pressure (p ≤ 0.05). The aortic valve extracellular matrix showed loss of elastin fibers and increased collagen deposition in the HF diet group. Collagen was quantified with ELISA, which showed an increased concentration of collagen types 1 and 3 (p ≤ 0.05). In the HF diet group, the initial stages of microcalcification were observed. Uniaxial mechanical testing of aortic cusps revealed that the HF diet group expressed a decrease in ultimate tensile strength and elastic modulus compared to the control diet group (p ≤ 0.05). Western blot and immunohistochemistry indicated the presence of proteins: lipoprotein-associated phospholipase A2, osteopontin, and osteocalcin with an increased expression in the HF diet group. The current study demonstrates that experimental metabolic syndrome induced by a 16-week HF diet was associated with a statistically significant alteration to the physical architecture of the aortic valve.


A critical review of time-weighted average as an index of exposure and dose, and of its key elements.
Time-weighted average (TWA) is widely used in research and practice, in occupational health, as an index of exposure and dose. Its key element, CT, where C is concentration of contaminant and T is duration of contamination, is recognizable as Haber's rule. Neither TWA nor similar measurements have been scientifically validated, and it does not seem appropriately named. In the late 1940s and early 1950s, cautions were expressed about the scientific validity of TWA, but the specific scientific study of it did not begin until 1981. In the interim, the cautions appear to have been largely ignored. In 1985, TWA cannot be said to be scientifically valid. Uses of it often confuse dose and exposure, and take insufficient account of time as a factor. Validation of TWA may well be too complicated a scientific problem for occupational epidemiology. TWA need not be rejected for regulatory or all practical purposes, but its key element ought to be better understood, and all applications which presume its scientific validity need to be reviewed.


A heterogeneous population of defective RNAs is associated with lettuce infectious yellows virus.
Preparations of dsRNAs and virion RNAs extracted from Nicotiana clevelandii plants infected with the bipartite Lettuce infectious yellows virus (LIYV) were found to contain multiple LIYV RNA species. In addition to the two LIYV genomic RNAs, three types of RNAs were observed: (a) 3' coterminal subgenomic RNAs; (b) RNAs containing LIYV RNA 1 or RNA 2 5' terminus but lacking the 3' terminus; and (c) RNAs with both LIYV RNA 2 3' and 5' termini but each with a central extensive deletion, a structure typical of defective RNAs (D RNAs). No D RNA-like RNAs were detected for LIYV RNA 1. A reverse transcription followed by polymerase chain reaction (RT-PCR) strategy was used to clone from virion RNAs several LIYV RNA 2 D RNAs as cDNAs. Nucleotide sequence analysis of 43 cloned cDNAs showed in some D RNAs the presence of a stretch of 1-5 nt in the junction site that is repeated in the genomic RNA 2 in the two positions flanking the junction site or in close proximity. Some D RNAs contained in the junction site one or several extra nucleotides not present in the LIYV genomic RNA 2. Two of the cloned cDNAs were used to generate in vitro transcripts, and infectivity studies showed that both D RNAs were replication competent in protoplasts when coinoculated with LIYV RNAs 1 and 2 or with only LIYV RNA1. Neither D RNA showed obvious effects upon LIYV RNA 1 and RNA 2 accumulation in coinfected protoplasts. These data suggest that LIYV infections contain a heterogeneous population of LIYV RNA 2 D RNAs, and some are encapsidated into virions.


Developing a woman-centered, inclusive definition of traumatic childbirth experiences: A discussion paper.
Many women experience giving birth as traumatic. Although women's subjective experiences of trauma are considered the most important, currently there is no clear inclusive definition of a traumatic birth to help guide practice, education, and research. To formulate a woman-centered, inclusive definition of a traumatic childbirth experience. After a rapid literature review, a five-step process was undertaken. First, a draft definition was created based on interdisciplinary experts' views. The definition was then discussed and reformulated with input from over 60 multidisciplinary clinicians and researchers during a perinatal mental health and birth trauma research meeting in Europe. A revised definition was then shared with consumer groups in eight countries to confirm its face validity and adjusted based on their feedback. The stepwise process confirmed that a woman-centered and inclusive definition was important. The final definition was: "A traumatic childbirth experience refers to a woman's experience of interactions and/or events directly related to childbirth that caused overwhelming distressing emotions and reactions; leading to short and/ or long-term negative impacts on a woman's health and wellbeing." This definition of a traumatic childbirth experience was developed through consultations with experts and consumer groups. The definition acknowledges that low-quality provider interactions and obstetric violence can traumatize individuals during childbirth. The women-centered and inclusive focus could help women to identify and validate their experiences of traumatic birth, offering benefits for practice, education, and research, as well as for policymaking and activism in the fields of perinatal mental health and respectful maternity care.


Integration of B-type natriuretic Peptide in heart failure outpatient programs.
In many European countries, patients with heart failure (HF) are treated according to specific management programs. The key players are the HF cardiologists and the HF nurses, and they are supported by a number of other important players. The objectives of the HF team are to help verify the diagnosis, optimize treatment, and inform and educate the patients and their relatives to achieve beneficial effects in terms of improved survival, less hospitalization, and improved quality of life. The use of natriuretic peptides (NPs) helps the referring physician correctly diagnose and select patients for further investigations. Based on the levels of NPs, it is possible to risk-stratify patients and offer them individualized, tailored treatment. The present paper discusses and exemplifies how NP assessment may be integrated into an HF outpatient program. Our conclusion is that NP values will continue to be important and necessary tools in the routine management of patients with HF.