Influence of light-curing mode on the cytotoxicity of resin-based surface sealants.
Surface sealants have been successfully used in the prevention of erosive tooth wear. However, when multiple tooth surfaces should be sealed, the light-curing procedure is very time-consuming. Therefore, the aim of this study was to investigate whether reduced light-curing time (while maintaining similar energy density) has an influence on resin-based surface sealant cytotoxicity. Bovine dentine discs were treated as follows: group 1: untreated, groups 2-5: Seal&Protect and groups 6-9: experimental sealer. Groups 2 and 6 were light-cured (VALO LED light-curing device) for 40 s (1000 mW/cm2), groups 3 and 7 for 10 s (1000 mW/cm2), groups 4 and 8 for 7 s (1400 mW/cm2) and groups 5 and 9 for 3 s (3200 mW/cm2). Later, materials were extracted in culture medium for 24 h, and released lactate dehydrogenase (LDH) activity as a measure of cytotoxicity was determined photometrically after cells (dental pulp cells and gingival fibroblasts) were exposed to the extracts for 24 h. Three independent experiments, for both sample preparation and cytotoxicity testing, were performed. Overall, lowest cytotoxicity was observed for the unsealed control group. No significant influence of light-curing settings on the cytotoxicity was observed (p = 0.537 and 0.838 for pulp cells and gingival fibroblasts, respectively). No significant difference in the cytotoxicity of the two sealants was observed after light-curing with same light-curing settings (group 2 vs. 6, 3 vs. 7, 4 vs. 8 and 5 vs. 9: p > 0.05, respectively). Shortening the light-curing time, while maintaining constant energy density, resulted in no higher cytotoxicity of the investigated sealants.


[Outpatient expectations in dermatology consultations: A pilot study].
Understanding patients' expectations with regard to medical care is critical as it guarantees an efficient therapeutic process. Our aim was to determine outpatients' expectations concerning clinical encounters in a dermatology clinic and to study how these matched the opinions of dermatologists regarding them. Consecutive outpatients consulting in five dermatology centres in the Paris suburbs between February 2013 and March 2013 were prospectively included. For this pilot cross-sectional study, we used two standardized forms to collect data from patients and dermatologists. Patients' answers were compared to those of their dermatologist, and the degree of matching was calculated to assess the ability of dermatologists to accurately identify their patients' expectations. Two hundred and sixty-five patients were included, with a median age of 41 years (interquartile range: 25; 62), of whom 166 were women (65.4%). Patient's principal expectations concerned diagnosis (51.7%) and medication (40.8%), with 32.1% of patients requiring reassurance. The rates of matching between patients' and dermatologists' answers ranged from 33.3% to 65.7% according to the type of expectations. The highest rate concerned expectation with regard to medications, being only 52.6% and 58.8%, respectively for expectations regarding diagnosis and the need for reassurance. This study highlights the need for improved identification of outpatient expectations in dermatology consultations.


MFS Transporters and GABA Metabolism Are Involved in the Self-Defense Against DON in Fusarium graminearum.
Trichothecene mycotoxins, such as deoxynivalenol (DON) produced by the fungal pathogen, Fusarium graminearum, are not only important for plant infection but are also harmful to human and animal health. Trichothecene targets the ribosomal protein Rpl3 that is conserved in eukaryotes. Hence, a self-defense mechanism must exist in DON-producing fungi. It is reported that TRI (trichothecene biosynthesis) 101 and TRI12 are two genes responsible for self-defense against trichothecene toxins in Fusarium. In this study, however, we found that simultaneous disruption of TRI101 and TRI12 has no obvious influence on DON resistance upon exogenous DON treatment in F. graminearum, suggesting that other mechanisms may be involved in self-defense. By using RNA-seq, we identified 253 genes specifically induced in DON-treated cultures compared with samples from cultures treated or untreated with cycloheximide, a commonly used inhibitor of eukaryotic protein synthesis. We found that transporter genes are significantly enriched in this group of DON-induced genes. Of those genes, 15 encode major facilitator superfamily transporters likely involved in mycotoxin efflux. Significantly, we found that genes involved in the metabolism of gamma-aminobutyric acid (GABA), a known inducer of DON production in F. graminearum, are significantly enriched among the DON-induced genes. The GABA biosynthesis gene PROLINE UTILIZATION 2-2 (PUT2-2) is downregulated, while GABA degradation genes are upregulated at least twofold upon treatment with DON, resulting in decreased levels of GABA. Taken together, our results suggest that transporters influencing DON efflux are important for self-defense and that GABA mediates the balance of DON production and self-defense in F. graminearum.


Intra-uterine growth restriction and increased risk of hypertension in adult life: a follow-up study of 50-year-olds.
To compare blood pressure between 50-year-old adults who were born at term (37-42 weeks of gestation) with intra-uterine growth restriction (IUGR; birth weight <10th centile) and a control group of similar age born at term without IUGR (birth weight ≥10th centile). Controlled comparative study. Participants included 232 men and women who were born at the Royal Maternity Hospital, Belfast, a large regional maternity hospital in Northern Ireland, between 1954 and 1956. One hundred and eight subjects who were born with IUGR were compared with 124 controls with normal birth weight for gestation. The main outcome measures were systolic and diastolic blood pressure at approximately 50 years of age, measured according to European recommendations. The IUGR group had higher systolic and diastolic blood pressure than the control group: 131.5 [95% confidence interval (CI) 127.9-135.1] vs 127.1 (95% CI 124.3-129.2) mmHg and 82.3 (95% CI 79.6-85.0) vs 79.0 (95% CI 77.0-81.0) mmHg, respectively. After adjustment for gender, the differences between the groups were statistically significant: systolic blood pressure 4.5 (95% CI 0.3-8.7) mmHg and diastolic blood pressure 3.4 (95% CI 0.2-6.5) mmHg (both P < 0.05). More participants in the IUGR group were receiving treatment for high blood pressure compared with the control group [16 (15%) vs 11 (9%)], although this was not statistically significant. The proportion of subjects with blood pressure >140/90 mmHg or currently receiving antihypertensive treatment was 45% (n = 49) for the IUGR group, and 31% (n = 38) for the control group (odds ratio 1.9, 95% CI 1.1-3.3). Adjustment for potential confounders made little difference. IUGR is associated with higher blood pressure at 50 years of age. Individuals born with IUGR should have regular blood pressure screening and early treatment as required. Hypertension remains underdiagnosed and undertreated in adult life.


Theoretical studies on Rh(iii)-catalyzed regioselective C-H bond cyanation of indole and indoline.
Density functional theory calculations were carried out to study the reaction mechanism of the Rh(iii)-catalyzed regioselective C-H cyanation of indole and indoline with N-cyano-N-phenyl-para-methylbenzenesulfonamide (NCTS). This mechanism involves four major steps: C-H activation, cyano group insertion, β-N elimination, and regeneration of active species. How different indole and indoline substrates affect the regioselectivity of C-H bond cyanation has been examined and analyzed in detail. Our calculation results indicate that the regioselectivity of C-H bond cyanation of indole depends on the nucleophilicity of carbon atoms in C-Rh(iii) bonds to the cyano group. For indoline, it can be attributed to the different hybridization platforms of the C-H bond activation.


Electrical readout of protein microarrays on regular glass slides.
A new approach for the electrical readout of microarrays prepared on regular glass slides, using an array of impedimetric transducers (interdigitated electrodes, IDEs) is presented in this work. Impedance detection relies on the use of a urease-labeled immunoassay scheme. Urease is able to produce an increase in conductivity by hydrolysis of the urea substrate, which is measured with the IDEs and directly related to the amount of target analyte. Unlike previous electrical microarrays, the assay does not take place on top of the transducers but on a regular glass slide, which may enable the development of compact multiplexed analytical systems with lower cost per assay. A droplet of solution with the enzymatic substrate is deposited on each transducer of the array, and the microarray is positioned at a short distance (300 μm) so that each droplet wets one transducer and one spot of the microarray. This procedure allows reusing the transducer array for readout of a virtually unlimited number of microarrays. A microarray based on an immunoassay for the detection of a mouse generic protein in a concentration range from 0.03 to 30 μg mL(-1) was carried out to assess the performance of the electrical readout approach. A sigmoid response with a limit of detection of 0.1 μg mL(-1) and a dynamic range of 1 order of magnitude was obtained. A comparative study was also carried out with two well established analytical procedures. First, the urease-based immunoassay was tested in a 96 well microtiter plate using phenol red pH indicator and absorbance detection. Second, the microarray was carried out using the same target protein concentration range but applying a Cy3 label and fluorescence detection. Both assays allowed for the validation of the performance of the presented electrical readout system.


Analytical fitting model for rough-surface BRDF.
A physics-based model is developed for rough surface BRDF, taking into account angles of incidence and scattering, effective index, surface autocovariance, and correlation length. Shadowing is introduced on surface correlation length and reflectance. Separate terms are included for surface scatter, bulk scatter and retroreflection. Using the FindFit function in Mathematica, the functional form is fitted to BRDF measurements over a wide range of incident angles. The model has fourteen fitting parameters; once these are fixed, the model accurately describes scattering data over two orders of magnitude in BRDF without further adjustment. The resulting analytical model is convenient for numerical computations.


Effects of aging on circadian patterns of gene expression in the human prefrontal cortex.
With aging, significant changes in circadian rhythms occur, including a shift in phase toward a "morning" chronotype and a loss of rhythmicity in circulating hormones. However, the effects of aging on molecular rhythms in the human brain have remained elusive. Here, we used a previously described time-of-death analysis to identify transcripts throughout the genome that have a significant circadian rhythm in expression in the human prefrontal cortex [Brodmann's area 11 (BA11) and BA47]. Expression levels were determined by microarray analysis in 146 individuals. Rhythmicity in expression was found in ∼ 10% of detected transcripts (P < 0.05). Using a metaanalysis across the two brain areas, we identified a core set of 235 genes (q < 0.05) with significant circadian rhythms of expression. These 235 genes showed 92% concordance in the phase of expression between the two areas. In addition to the canonical core circadian genes, a number of other genes were found to exhibit rhythmic expression in the brain. Notably, we identified more than 1,000 genes (1,186 in BA11; 1,591 in BA47) that exhibited age-dependent rhythmicity or alterations in rhythmicity patterns with aging. Interestingly, a set of transcripts gained rhythmicity in older individuals, which may represent a compensatory mechanism due to a loss of canonical clock function. Thus, we confirm that rhythmic gene expression can be reliably measured in human brain and identified for the first time (to our knowledge) significant changes in molecular rhythms with aging that may contribute to altered cognition, sleep, and mood in later life.


Familial and socio-economic correlates of somatisation disorder.
Somatisation disorder can result from an interplay between suboptimal family environment and socio-economic deprivation, which enhances the underlying cognitive tendency for this disorder. There are pertinent familial and socio-economic factors associated with this disorder, but research addressing this is sparse. AIM AND SETTING: The study aims to evaluate family and socio-economic factors that are associated with somatisation disorder amongst patients presenting to the Family Medicine clinic, University College Hospital, Ibadan, Nigeria. This is an observational case-control study of 120 participants who presented to the clinic between May and August 2009. Data collection was by interviewer-administered structured questionnaire using the World Health Organization Screener for Somatoform Disorder and Somatoform Disorder Schedule to ascertain somatisation in 60 patients who were then matched with 60 controls. The respondents' demographic and family data were also collected and their interpersonal relationships were assessed with the Family Relationship Index. The somatising patients were mostly females (70%), with a female to male ratio of 2.3:1 and mean age of 43.65 ± 13.04 years.Living in a polygamous family (as any member of the family) was significantly related to somatisation (p = 0.04). Somatisation was also more common in people who were separated, divorced or widowed (p = 0.039). Somatisers from a lower social class or those earning below a dollar a day experienced poorer cohesion (p = 0.042) and more conflicts (p = 0.019) in their interpersonal relationship. This study was able to demonstrate that a polygamous family setting, disrupted marriage, low social status and financial constraints are correlates of somatisation. It is of essence to identify these factors in holistic management of somatising patients.


Thyroid stimulating hormone and cognition during severe, transient hypothyroidism.
The purpose of our pilot study was to explore the relationship between serum thyroid stimulating hormone (TSH) levels during overt hypothyroidism (OH) and hypothyroid-related neuropsychological symptoms. We hypothesized that TSH level may reflect the degree of 'brain hypothyroidism' such that an inverse correlation may exist between serum TSH and cognitive function in patients experiencing overt hypothyroidism (OH), and sought to explore this hypothesis. Eleven thyroidectomized patients underwent neuropsychological and thyroid function testing while overtly hypothyroid, and again following thyroid hormone replacement. Their test performance was compared with that of eleven healthy controls at a similarly separated two points in time, and the change over time for the patient group and the controls was likewise assessed and compared. The patients' neuropsychological test scores were then correlated with their serum TSH levels while hypothyroid. The patients' performance while hypothyroid was worse than that of the controls in only one neurocognitive measure--Working Memory Index. The subjects improved similarly or to a greater degree than the controls, when the subjects were thyroid hormone replaced, on all but one neurocognitive measure - Thurstone Word Fluency. TSH level during hypothyroidism was inversely proportional to the patients' performance on these same two measures, but no others. Serum TSH level during hypothyroidism was inversely proportional to performance on the only two neurocognitive measures evidencing an adverse effect from hypothyroidism in our cohort. This suggests that serum TSH level may reflect the severity of 'brain hypothyroidism' during the overt stage of this condition.


Single-cell analyses of transcriptional heterogeneity during drug tolerance transition in cancer cells by RNA sequencing.
The acute cellular response to stress generates a subpopulation of reversibly stress-tolerant cells under conditions that are lethal to the majority of the population. Stress tolerance is attributed to heterogeneity of gene expression within the population to ensure survival of a minority. We performed whole transcriptome sequencing analyses of metastatic human breast cancer cells subjected to the chemotherapeutic agent paclitaxel at the single-cell and population levels. Here we show that specific transcriptional programs are enacted within untreated, stressed, and drug-tolerant cell groups while generating high heterogeneity between single cells within and between groups. We further demonstrate that drug-tolerant cells contain specific RNA variants residing in genes involved in microtubule organization and stabilization, as well as cell adhesion and cell surface signaling. In addition, the gene expression profile of drug-tolerant cells is similar to that of untreated cells within a few doublings. Thus, single-cell analyses reveal the dynamics of the stress response in terms of cell-specific RNA variants driving heterogeneity, the survival of a minority population through generation of specific RNA variants, and the efficient reconversion of stress-tolerant cells back to normalcy.


Effects of water and feed restriction on body weight change and nitrogen balance in desert goats fed high and low quality forages.
The effects of water and food restriction on feed utilization, body weight changes and nitrogen balance were investigated in Sudanese desert goats given high or low quality forage. Nine male goats were employed in a 3x3 Latin square design where they were subjected to three treatments: ad libitum water and feed, ad-libitum feed/restricted water, and ad libitum water/restricted feed. These treatments were repeated using lucerne or sorghum hay. For both feed types dry matter intake (DMI) was not affected by treatments. Water intake decreased with feed restriction in goats fed sorghum hay. The ratio of water intake to DMI increased with lucerne hay compared to sorghum hay in both the control and feed restricted group. Water consumption increased with lucerne compared to sorghum hay. Body weight losses were more pronounced with water than with feed restriction but were less severe with lucerne than sorghum hay. Interactions due to treatment and feed type were significant for water intake and body weight change. With lucerne hay, except for crude protein (CP), nutrient digestibility improved with water restriction. Feeding sorghum hay, there were no effects on digestibility except for crude protein (CP) that was reduced with water restriction. Crude fiber (CF) and nitrogen free extract (NFE) digestibility increased with feed restriction in animals on sorghum hay. For all groups, CP digestibility was better with lucerne than sorghum hay. Interactions of the main effects were significant for all apparent digestibility coefficients as well as total digestible nutrients. The treatments did not have significant effects on nitrogen balance, however, nitrogen intake, retained or excreted in faeces was higher with lucerne than sorghum hay. It may be concluded that subjecting animals to water or feed restriction will have a more deleterious effect on feed utilization and nitrogen balance when using a low than a high quality forage.


Modeling cell gradient sensing and migration in competing chemoattractant fields.
Directed cell migration mediates physiological and pathological processes. In particular, immune cell trafficking in tissues is crucial for inducing immune responses and is coordinated by multiple environmental cues such as chemoattractant gradients. Although the chemotaxis mechanism has been extensively studied, how cells integrate multiple chemotactic signals for effective trafficking and positioning in tissues is not clearly defined. Results from previous neutrophil chemotaxis experiments and modeling studies suggested that ligand-induced homologous receptor desensitization may provide an important mechanism for cell migration in competing chemoattractant gradients. However, the previous mathematical model is oversimplified to cell gradient sensing in one-dimensional (1-D) environment. To better understand the receptor desensitization mechanism for chemotactic navigation, we further developed the model to test the role of homologous receptor desensitization in regulating both cell gradient sensing and migration in different configurations of chemoattractant fields in two-dimension (2-D). Our results show that cells expressing normal desensitizable receptors preferentially orient and migrate toward the distant gradient in the presence of a second local competing gradient, which are consistent with the experimentally observed preferential migration of cells toward the distant attractant source and confirm the requirement of receptor desensitization for such migratory behaviors. Furthermore, our results are in qualitative agreement with the experimentally observed cell migration patterns in different configurations of competing chemoattractant fields.


Impact of atrial fibrillation on clinical outcomes among patients with coronary artery disease undergoing revascularisation with drug-eluting stents.
Coronary artery disease (CAD) and atrial fibrillation (AF) are major determinants of morbidity and mortality. A combined treatment with antiplatelet agents and vitamin K antagonists limits the risk of stent thrombosis and stroke while increasing the rate of bleeding. The objective of this study was to investigate the impact of atrial fibrillation (AF) on long-term clinical outcomes in patients with CAD undergoing percutaneous coronary intervention (PCI) with drug-eluting stents (DES). Among 6,308 consecutive patients undergoing PCI with DES between 2002 and 2009, 323 (5.3%) patients were diagnosed with AF. We compared clinical outcomes between patients with and those without AF throughout four years. The primary endpoint was a composite of all-cause mortality, myocardial infarction (MI), ischaemic stroke, and BARC bleeding type 3b/3c/5a/5b. In adjusted analyses, the primary composite endpoint was more frequent among patients with AF (HR 1.59, 95% CI 1.26-2.00; p<0.001). Differences were driven by an increased risk of all-cause mortality (HR 1.67, 95% CI 1.27-2.20; p=0.003), ischaemic stroke (HR 3.09, 95% CI 1.45-6.56; p=0.003), and intracranial bleeding (HR 4.28, 95% CI 1.36-13.48; p=0.013). We observed a gradient of risk among patients with higher CHA2DS2-VASc scores and modified outpatient bleeding risk index. Among patients with CAD undergoing revascularisation with DES, AF confers an increased risk of all-cause mortality, ischaemic stroke, and intracranial bleeding. The hazard imposed by AF correlates with the CHA2DS2-VASc score.


Expression of Neuroendocrine Factor VGF in Lung Cancer Cells Confers Resistance to EGFR Kinase Inhibitors and Triggers Epithelial-to-Mesenchymal Transition.
Mutations in EGFR drive tumor growth but render tumor cells sensitive to treatment with EGFR tyrosine kinase inhibitors (TKI). Phenotypic alteration in epithelial-to-mesenchymal transition (EMT) has been linked to the TKI resistance in lung adenocarcinoma. However, the mechanism underlying this resistance remains unclear. Here we report that high expression of a neuroendocrine factor termed VGF induces the transcription factor TWIST1 to facilitate TKI resistance, EMT, and cancer dissemination in a subset of lung adenocarcinoma cells. VGF silencing resensitized EGFR-mutated lung adenocarcinoma cells to TKI. Conversely, overexpression of VGF in sensitive cells conferred resistance to TKIs and induced EMT, increasing migratory and invasive behaviors. Correlation analysis revealed a significant association of VGF expression with advanced tumor grade and poor survival in patients with lung adenocarcinoma. In a mouse xenograft model of lung adenocarcinoma, suppressing VGF expression was sufficient to attenuate tumor growth. Overall, our findings show how VGF can confer TKI resistance and trigger EMT, suggesting its potential utility as a biomarker and therapeutic target in lung adenocarcinoma. Cancer Res; 77(11); 3013-26. ©2017 AACR.


Goosecoid and mix.1 repress Brachyury expression and are required for head formation in Xenopus.
The Xenopus homologue of Brachyury, Xbra, is expressed in the presumptive mesoderm of the early gastrula. Induction of Xbra in animal pole tissue by activin occurs only in a narrow window of activin concentrations; if the level of inducer is too high, or too low, the gene is not expressed. Previously, we have suggested that the suppression of Xbra by high concentrations of activin is due to the action of genes such as goosecoid and Mix.1. Here, we examine the roles played by goosecoid and Mix.1 during normal development, first in the control of Xbra expression and then in the formation of the mesendoderm. Consistent with the model outlined above, inhibition of the function of either gene product leads to transient ectopic expression of Xbra. Such embryos later develop dorsoanterior defects and, in the case of interference with Mix.1, additional defects in heart and gut formation. Goosecoid, a transcriptional repressor, appears to act directly on transcription of Xbra. In contrast, Mix.1, which functions as a transcriptional activator, may act on Xbra indirectly, in part through activation of goosecoid.


Bayesian Data Assimilation to Support Informed Decision Making in Individualized Chemotherapy.
An essential component of therapeutic drug/biomarker monitoring (TDM) is to combine patient data with prior knowledge for model-based predictions of therapy outcomes. Current Bayesian forecasting tools typically rely only on the most probable model parameters (maximum a posteriori (MAP) estimate). This MAP-based approach, however, does neither necessarily predict the most probable outcome nor does it quantify the risks of treatment inefficacy or toxicity. Bayesian data assimilation (DA) methods overcome these limitations by providing a comprehensive uncertainty quantification. We compare DA methods with MAP-based approaches and show how probabilistic statements about key markers related to chemotherapy-induced neutropenia can be leveraged for more informative decision support in individualized chemotherapy. Sequential Bayesian DA proved to be most computationally efficient for handling interoccasion variability and integrating TDM data. For new digital monitoring devices enabling more frequent data collection, these features will be of critical importance to improve patient care decisions in various therapeutic areas.


Noise exposure immediately activates cochlear mitogen-activated protein kinase signaling.
Noise-induced hearing loss (NIHL) is a major public health issue worldwide. Uncovering the early molecular events associated with NIHL would reveal mechanisms leading to the hearing loss. Our aim is to investigate the immediate molecular responses after different levels of noise exposure and identify the common and distinct pathways that mediate NIHL. Previous work showed mice exposed to 116 decibels sound pressure level (dB SPL) broadband noise for 1 h had greater threshold shifts than the mice exposed to 110 dB SPL broadband noise, hence we used these two noise levels in this study. Groups of 4-8-week-old CBA/CaJ mice were exposed to no noise (control) or to broadband noise for 1 h, followed by transcriptome analysis of total cochlear RNA isolated immediately after noise exposure. Previously identified and novel genes were found in all data sets. Following exposure to noise at 116 dB SPL, the earliest responses included up-regulation of 243 genes and down-regulation of 61 genes, while a similar exposure at 110 dB SPL up-regulated 155 genes and down-regulated 221 genes. Bioinformatics analysis indicated that mitogen-activated protein kinase (MAPK) signaling was the major pathway in both levels of noise exposure. Nevertheless, both qualitative and quantitative differences were noticed in some MAPK signaling genes, after exposure to different noise levels. Cacna1b , Cacna1g , and Pla2g6 , related to calcium signaling were down-regulated after 110 dB SPL exposure, while the fold increase in the expression of Fos was relatively lower than what was observed after 116 dB SPL exposure. These subtle variations provide insight on the factors that may contribute to the differences in NIHL despite the activation of a common pathway.


Nonfunctional variant 3 factor H binding proteins as meningococcal vaccine candidates.
Neisseria meningitidis is a human-specific pathogen and leading cause of meningitis and septicemia. Factor H binding protein (fHbp), a virulence factor which protects N. meningitidis from innate immunity by binding the human complement regulator factor H (fH) with high affinity, is also a key antigen in vaccines being developed to prevent meningococcal disease. fHbp can be divided into three variant groups (V1, V2, and V3) that elicit limited immunological cross-reactivity. The interaction of fH with fHbp could impair the immunogenicity of this antigen by hindering access to the antigenic epitopes in fHbp, providing the rationale for the development of nonfunctional fHbps as vaccine candidates. Here, we characterized the two nonfunctional V3 fHbps, fHbp(T286A) and fHbp(E313A), which each contains a single amino acid substitution that leads to a marked reduction in affinity for fH without affecting the folding of the proteins. The immunogenicity of the nonfunctional fHbps was assessed in transgenic mice expressing a single chimeric fH containing domains of human fH involved in binding to fHbp. No differences in anti-V3 fHbp antibody titers were elicited by the wild-type V3 fHbp, V3 fHbp(T286A), and V3 fHbp(E313A), demonstrating that the nonfunctional fHbps retain their immunogenicity. Furthermore, the nonfunctional V3 fHbps elicit serum bactericidal activity that is equivalent to or higher than that observed with the wild-type protein. Our findings provide the basis for the rational design of next-generation vaccines containing nonfunctional V3 fHbps.


Blind prediction of cyclohexane-water distribution coefficients from the SAMPL5 challenge.
In the recent SAMPL5 challenge, participants submitted predictions for cyclohexane/water distribution coefficients for a set of 53 small molecules. Distribution coefficients (log D) replace the hydration free energies that were a central part of the past five SAMPL challenges. A wide variety of computational methods were represented by the 76 submissions from 18 participating groups. Here, we analyze submissions by a variety of error metrics and provide details for a number of reference calculations we performed. As in the SAMPL4 challenge, we assessed the ability of participants to evaluate not just their statistical uncertainty, but their model uncertainty-how well they can predict the magnitude of their model or force field error for specific predictions. Unfortunately, this remains an area where prediction and analysis need improvement. In SAMPL4 the top performing submissions achieved a root-mean-squared error (RMSE) around 1.5 kcal/mol. If we anticipate accuracy in log D predictions to be similar to the hydration free energy predictions in SAMPL4, the expected error here would be around 1.54 log units. Only a few submissions had an RMSE below 2.5 log units in their predicted log D values. However, distribution coefficients introduced complexities not present in past SAMPL challenges, including tautomer enumeration, that are likely to be important in predicting biomolecular properties of interest to drug discovery, therefore some decrease in accuracy would be expected. Overall, the SAMPL5 distribution coefficient challenge provided great insight into the importance of modeling a variety of physical effects. We believe these types of measurements will be a promising source of data for future blind challenges, especially in view of the relatively straightforward nature of the experiments and the level of insight provided.


Defining cardiovascular toxicities of cancer therapies: an International Cardio-Oncology Society (IC-OS) consensus statement.
The discipline of Cardio-Oncology has seen tremendous growth over the past decade. It is devoted to the cardiovascular (CV) care of the cancer patient, especially to the mitigation and management of CV complications or toxicities of cancer therapies, which can have profound implications on prognosis. To that effect, many studies have assessed CV toxicities in patients undergoing various types of cancer therapies; however, direct comparisons have proven difficult due to lack of uniformity in CV toxicity endpoints. Similarly, in clinical practice, there can be substantial differences in the understanding of what constitutes CV toxicity, which can lead to significant variation in patient management and outcomes. This document addresses these issues and provides consensus definitions for the most commonly reported CV toxicities, including cardiomyopathy/heart failure and myocarditis, vascular toxicity, and hypertension, as well as arrhythmias and QTc prolongation. The current document reflects a harmonizing review of the current landscape in CV toxicities and the definitions used to define these. This consensus effort aims to provide a structure for definitions of CV toxicity in the clinic and for future research. It will be important to link the definitions outlined herein to outcomes in clinical practice and CV endpoints in clinical trials. It should facilitate communication across various disciplines to improve clinical outcomes for cancer patients with CV diseases.


Influence of intrinsic microbes on phytochemical changes and antioxidant activity of the Ayurvedic fermented medicines: Balarishta and Chandanasava.
Balarishta and Chandanasava are polyherbal-fermented medicines of Ayurveda. Investigation of native microbes, understanding phytochemical changes and antioxidant activities in these medicines. Microbial populations were enumerated using selective media and standard plating methods. Yeast and bacteria were identified using classical and molecular methods. Qualitative phytochemical and gas chromatography-mass spectrometry (GC-MS) analyses were carried out. In vitro antioxidant assays were performed with different assay systems. Balarishta and Chandanasava possess two yeasts (Saccharomyces cerevisiae and Schizosaccharomyces pombe) and six bacteria that are species of Bacillus, Paenibacillus, and Brevibacillus. These microbes identified biochemically were authenticated with 16S and 18S rDNA sequence analysis and NCBI accession numbers. GC-MS analysis indicated that several compounds disappear as a result of fermentation while many are retained. The presence of new phytochemical compounds in the final stages of fermentation could be ascribed from the parent molecules that either disappeared or retained during fermentation. It suggests the biotransformation of phytochemicals by the mediation of intrinsic microbes. These medicines possess antioxidant activities by the presence of phytochemicals such as phenolics, flavonoids, tannins and phytosterols, wherein bacteria also contribute. The role of native microbial consortium in fermentation, biotransformation and antioxidant activity of these Arishta and Asava is demonstrated.


Hdac4 Interactions in Huntington's Disease Viewed Through the Prism of Multiomics.
Huntington's disease (HD) is a monogenic disorder, driven by the expansion of a trinucleotide (CAG) repeat within the huntingtin (Htt) gene and culminating in neuronal degeneration in the brain, predominantly in the striatum and cortex. Histone deacetylase 4 (Hdac4) was previously found to contribute to the disease progression, providing a potential therapeutic target. Hdac4 knockdown reduced accumulation of misfolded Htt protein and improved HD phenotypes. However, the underlying mechanism remains unclear, given its independence on deacetylase activity and the predominant cytoplasmic Hdac4 localization in the brain. Here, we undertook a multiomics approach to uncover the function of Hdac4 in the context of HD pathogenesis. We characterized the interactome of endogenous Hdac4 in brains of HD mouse models. Alterations in interactions were investigated in response to Htt polyQ length, comparing mice with normal (Q20) and disease (Q140) Htt, at both pre- and post-symptomatic ages (2 and 10 months, respectively). Parallel analyses for Hdac5, a related class IIa Hdac, highlighted the unique interaction network established by Hdac4. To validate and distinguish interactions specifically enhanced in an HD-vulnerable brain region, we next characterized endogenous Hdac4 interactions in dissected striata from this HD mouse series. Hdac4 associations were polyQ-dependent in the striatum, but not in the whole brain, particularly in symptomatic mice. Hdac5 interactions did not exhibit polyQ dependence. To identify which Hdac4 interactions and functions could participate in HD pathogenesis, we integrated our interactome with proteome and transcriptome data sets generated from the striata. We discovered an overlap in enriched functional classes with the Hdac4 interactome, particularly in vesicular trafficking and synaptic functions, and we further validated the Hdac4 interaction with the Wiskott-Aldrich Syndrome Protein and SCAR Homolog (WASH) complex. This study expands the knowledge of Hdac4 regulation and functions in HD, adding to the understanding of the molecular underpinning of HD phenotypes.


Structural changes in female-to-male sex-reversing gonads of Rana RUGOSA.
The phenotypic sex of many species of amphibians is subject to reversal by steroid hormones. The mechanism of this process, however, still remains largely unknown. As a step toward understanding the histological changes during sex reversal in amphibians, we analyzed two- and three-dimensional (2D and 3D) structures of sex-reversing gonads in Rana rugosa frogs. 2D views revealed that many oocytes in the wild-type ovary disappeared during female-to-male sex-reversal concomitant with the emergence of Vasa-positive small germ cells. Some of the germ cells were labeled with BrdU. BrdU-positive germ cells were few in the testosterone (T) treated ovaries at days 8 and 16, which resembled wild-type ovaries. Basement membranes became disrupted by day 24 in T-treated ovaries. However, the membranes were later reconfigured into testis-like gonadal structures 40 days after T treatment. 3D imaging of the sex-reversing gonad using serial immunostained sections showed that germ cells were organized in linear fashion extending out from where the sex-reversing gonad attached to the mesorchium 24 days after T treatment. Germ cells were increased in number by 40 days and were localized to the cortex of the gonads. In a T-untreated testis at day 24, many germ cells were distributed throughout the cortex except in the central space, while the efferent duct ran between two sheets of the mesorchium. These results, taken together, suggest that the mesorchium plays an important role in the organization of testicular structure. This is the first report showing germ cell ontogeny and organization in the female-to-male sex-reversing gonad in a vertebrate species.


Synergy of molecular and serological methods in minimally invasive diagnosis of enteroviral cardiac infection.
Treatment of myocarditis and pericarditis can differ on the basis of aetiology: systemic or auto-immune disease can be positively influenced by corticoid therapy, whereas this kind of treatment can worsen the course of virus-induced disease. Therefore, the aetiological diagnosis is extremely important. The synergistic use of minimally invasive serological, IgG, IgM, IgA, and neutralizing titres, and RNA detection was evaluated on representative patients out of 238 suffering from cardiopathies. The results obtained for each case can yield reliable guidelines that rapidly highlight the presence of a viral aetiology so that an endomyocardial biopsy can be performed thus eliminating incorrect therapies. Thus, not only is this technique rapid, minimally invasive providing the clinician with decisive data, but it is cost effective for the health system.


Immunoelectron microscope visualization of nuclear ribonucleoprotein antigens within spread transcription complexes.
The ultrastructural distribution of nuclear ribonucleoproteins (RNP) within spread active chromatin has been investigated using specific anti-RNP antibodies. Monoclonal antibodies directed against the core proteins of heterogeneous nuclear (hn)RNP or against small nuclear (sn)RNP have been incubated directly with lysed mouse or Drosophila tissue culture cells and the bound antibodies visualized by means of a protein A-colloidal gold complex. The hnRNP core proteins have been localized on growing RNP fibrils within non-nucleolar transcription complexes. Anti-snRNP antibodies, directed either against the Sm-antigen (common for nucleoplasmic snRNP species containing U1, U2, U4, U5, and U6 RNAs) or against U1-snRNP, were bound by two morphological types of RNP structures. Within areas of chromatin that do not completely disperse, labeling was observed on RNP-fibril gradient type structures or on groups of fibrogranular material. In the well dispersed regions containing individual nonribosomal transcription complexes, snRNP antigens were associated with growing RNP fibrils. Our results provide direct evidence for association of some U-snRNP species (including U1-snRNP) with extranucleolar RNA as early as during transcription elongation. In addition, the presence of core hnRNP proteins on the same type of nascent RNA transcripts has been confirmed.


The isolation of Kemerovo group orbiviruses and Uukuniemi group viruses of the family bunyaviridae from Ixodes uriae ticks from the Isle of May, Scotland.
Viruses isolated from ticks (Ixodes uriae) from a seabird colony on the Isle of May, Scotland, were shown by complement fixation tests to be related to the Uukuniemi and Kemerovo serogroups. Electron microscopic studies on the Uukuniemi viruses showed them to have a morphology characteristic of the bunyaviridae, and the Kemerovo group to be characteristic of orbiviruses. Separate isolates from the two serogroups were distinguished from each other by neutralization tests.


Abolition of spindle oscillations in thalamic neurons disconnected from nucleus reticularis thalami.
The effects of depriving thalamic relay and intralaminar nuclei from their reticularis thalami (RE) inputs were investigated in acute and chronic experiments on cat. In acutely prepared animals, two (frontal and parasagittal) thalamic transections were made; extracellular and intracellular recordings were performed in RE-disconnected thalamic nuclei. In chronic experiments, the RE nuclear complex was lesioned by means of kainic acid injections; the activity of RE-deprived thalamocortical neurons was extracellularly studied during wakefulness and synchronized sleep. Two features distinguish RE-deprived nuclei from normal thalamic nuclei: absence of spindle-wave rhythmicity and all-burst activity of neurons. The abolition of spindle-related rhythms (sequences of 7- to 14-Hz waves recurring periodically with a rhythm of 0.1-0.2 Hz) in RE-disconnected thalamic nuclei and ipsilateral neocortical areas contrasted with normal spindling rhythmicity in contralateral EEG leads. Spontaneously occurring, rhythmic, long-lasting inhibitory postsynaptic potentials (IPSPs), as observed in intact preparations, were no longer observed in RE-disconnected thalamic neurons. The remaining inhibitory events consisted of short-duration IPSPs. The possibility that RE nucleus is a pacemaker for spindling rhythms, imposing them through inhibitory projections to target thalamic areas, is supported by our concurrent experiments that indicate RE neurons preserve their rhythmicity after disconnection from their major (cortical and thalamic) input sources. RE-deprived thalamocortical neurons exclusively exhibit high-frequency spike bursts whose intrinsic structure is identical to that of intact thalamic relay cells. Instead of the spindle-related sequences of bursts seen in normal animals, the bursts of RE-disconnected thalamocortical neurons are single events, with a dramatic rhythmicity at 1-2 Hz. The presumed mechanism of this rhythmicity is the periodic activation of a low-threshold somatic conductance whose deinactivation is brought about by temporal integration of short-lasting IPSPs. It is known that high-frequency spike bursts of thalamic relay neurons result from hyperpolarization of cell membrane. We blocked the underlying inhibitory events by bicuculline and reversibly changed the all-burst activity of RE-disconnected neurons into a tonic mode. Since the only activity of RE-deprived thalamocortical neurons consists of burst discharges, we hypothesize that local-circuit GABAergic neurons are released from inhibition after RE disconnection or lesion.


Proteomics analysis of potential serum biomarkers for insulin resistance in patients with polycystic ovary syndrome.
The aim of the present study was to identify potential serum biomarkers for insulin resistance (IR) in patients with polycystic ovary syndrome (PCOS) by comparing the differences in serum protein expression levels between PCOS patients with and without IR. PCOS patients aged from 18 to 35 years were recruited at Guangdong Women and Children's Hospital from January, 2013 to February, 2014. A total of 218 PCOS patients were enrolled and divided into the insulin resistance (PCOS‑IR) and non‑insulin resistance (PCOS‑NIR) groups according to their homeostasis model assessment of insulin resistance. Two‑dimensional difference gel electrophoresis (2D‑DIGE) and matrix‑assisted laser desorption/ionization time‑of‑flight mass spectrometry (MALDI‑TOF‑MS/MS) techniques were used to identify differences in protein expression levels between the PCOS‑IR and PCOS‑NIR groups. The present study demonstrated that the total cholesterol (TCH), triglycerides (TG), low‑density lipoprotein (LDL), fasting plasma glucose (FPG), 3‑h blood glucose (3hBG) and uric acid (UA) levels in the PCOS‑IR group were higher than those in the PCOS‑NIR group (P<0.05). Between the PCOS‑IR and PCOS‑NIR groups, a total of 20 differentially expressed protein spots were detected by 2D‑DIGE. Among these, 4 proteins, namely afamin, serotransferrin, complement C3 and apolipoprotein C3 (APOC3), were also identified by MALDI‑TOF‑MS/MS. The alteration of APOC3 was further confirmed by western blot analysis and enzyme‑linked immunosorbent assay (ELISA). The present study also confirmed that the expression level of APOC3 was positively associated with the homeostasis model assessment of insulin resistance (HOMA‑IR). On the whole, the data indicate that APOC3 may be a potential diagnostic marker for PCOS‑IR patients.


The prevention of ankle sprains in sports. A systematic review of the literature.
To assess the published evidence on the effectiveness of various approaches to the prevention of ankle sprains in athletes, we used textbooks, journals, and experts in the field of sports medicine to identify citations. We identified 113 studies reporting the risk of ankle sprains in sports, methods to provide support, the effect of these interventions on performance, and comparison of prevention efforts. The most common risk factor for ankle sprain in sports is history of a previous sprain. Ten citations of studies involving athletes in basketball, football, soccer, or volleyball compared alternative methods of prevention. Methods tested included wrapping the ankle with tape or cloth, orthoses, high-top shoes, or some combination of these methods. Most studies indicate that appropriately applied braces, tape, or orthoses do not adversely affect performance. Based on our review, we recommend that athletes with a sprained ankle complete supervised rehabilitation before returning to practice or competition, and those athletes suffering a moderate or severe sprain should wear an appropriate orthosis for at least 6 months. Both coaches and players must assume responsibility for prevention of injuries in sports. Methodologic limitations of published studies suggested several areas for future research.


Evaluation of different glycoforms of honeybee venom major allergen phospholipase A2 (Api m 1) produced in insect cells.
Allergic reactions to hymenoptera stings are one of the major reasons for IgE-mediated anaphylaxis. However, proper diagnosis using venom extracts is severely affected by molecular cross-reactivity. In this study recombinant honeybee venom major allergen phospholipase A2 (Api m 1) was produced for the first time in insect cells. Using baculovirus infection of different insect cell lines allergen versions providing a varying degree of cross-reactive carbohydrate determinants as well as a non glycosylated variant could be obtained as secreted soluble proteins in high yields. The resulting molecules were analyzed for their glycosylation and proved to show advantageous properties regarding cross-reactivity in sIgE-based assays. Additionally, in contrast to the enzymatically active native protein the inactivated allergen did not induce IgE-independent effector cell activation. Thus, insect cell-derived recombinant Api m 1 with defined CCD phenotypes might provide further insights into hymenoptera venom IgE reactivities and contribute to an improved diagnosis of hymenoptera venom allergy.


Diagnostic value of computerized tomography venography in detecting stenosis and occlusion of subclavian vein and superior vena in chronic renal failure patients.
Currently, venography is the standard diagnostic method to examine veins before implementing access, which is invasive in nature. Computerized tomography venography (CTV) can simultaneously indicate deep and superficial venous systems in the upper extremity and their relation to the surrounding anatomical structures; however, its diagnostic value in the detection of central venous disease has yet to be defined. The aim of this study was to determine the diagnostic value of CT venography compared to venography in the diagnosis of stenosis and the occlusion of subclavian veins and the superior vena in renal failure patients. This cross-sectional study was conducted from January to September 2015 on patients with chronic renal failure undergoing upper extremity venography at the Radiology Department of Imam Reza Hospital in Mashhad, Iran. We excluded patients with catheters in their jugular and subclavian vein routes, venous hypertension with reverse-function fistula, or sensitivity to contrast agents. Several factors, including age, gender, catheterization record in jugular and subclavian veins, and fistula record in the upper extremity, as well as clinical symptoms consisting of edema, dermatitis, and ulcers in these organs, were recorded in the corresponding form. Then, the patients consecutively underwent indirect venography and CT venography and traces of stenosis (more than 50%) or complete occlusion in the subclavian vein and superior vena were recorded. The data were analyzed using SPSS software by the chi-squared test, and sensitivity, specificity, and positive and negative predictive values were calculated by means of MedCalc Online, version 16.2. The study was conducted on 40 patients (26 males and 14 females) with a mean age of 46.7 ± 10.4 years. In this study, 58 subclavian veins, as well as 32 superior vena cava, were studied. The results showed that the diagnostic value of CTV in the detection of subclavian stenosis had a sensitivity and a specificity of 88.2 and 97.5%, respectively. Moreover, in the superior vena cava, the greatest CTV diagnostic sensitivity and specificity was related to vein stenosis detection (sensitivity = 88.8%; specificity = 100%). It seems that the CTV, based on its high sensitivity, specificity, and predictive value, can be used as an efficient tool in the study of stenosis in subclavian veins and superior vena in patients undergoing venography.


Comparison of Enrichment Methods for Intact N- and O-Linked Glycopeptides Using Strong Anion Exchange and Hydrophilic Interaction Liquid Chromatography.
Heterogeneity of protein glycosylation poses great challenges for analysis that is key to understand structure and function of glycoproteins. Resolving this conundrum requires efficient and specific enrichment of intact glycopeptides for identification and quantitation. To this end, hydrophilic interaction chromatography (HILIC) has been commonly used to enrich intact N- and O-linked glycopeptides. However, its effectiveness to enrich isobarically labeled glycopeptides remains unclear. Here, we studied three different enrichment methods for enrichment of N- and O-linked glycopeptides. It was found that removal of N-glycans prior to enrichment of O-linked glycopeptides by HILIC improved identification of O-linked glycopeptides by mass spectrometry. We also compared the enrichment of intact N- and O-linked glycopeptides using other chromatography methods and found that using cartridges containing materials for strong anion exchange (SAX) chromatography increased yield and identification of N- and O-linked glycopeptides. The enrichment of O-linked glycopeptides was further improved when a Retain AX cartridge (RAX) was used. In particular, isobaric tag labeled glycopeptides after C18 desalting could be readily enriched by SAX and RAX cartridges but not by HILIC to enable quantitative glycoproteomics. It is anticipated that the use of SAX and RAX cartridges will facilitate broad applications of identifications and quantitation of glycoproteins.


Modulation of response timing in ADHD, effects of reinforcement valence and magnitude.
The present study investigated the impact of reinforcement valence and magnitude on response timing in children with ADHD. Children were required to estimate a 1-s interval, and both the median response time (response tendency) and the intrasubject-variability (response stability) were investigated. In addition, heart rate and skin conductance were measured to examine the autonomic responses to reinforcement. Feedback-only trials were compared to low response cost trials (response cost for incorrect responses), low reward trials (reward for correct responses), high response cost and high reward trials. In feedback-only trials, children with ADHD underestimated more severely the interval and responded more variably than controls. Children with ADHD, unlike controls, were unaffected by the reinforcement conditions in terms of time underestimations. The variability of responding, on the other hand, decreased under conditions of reinforcement to a larger extent in children with ADHD than controls. There were no indications that children with ADHD were abnormally affected by the valence or magnitude of reinforcement. Furthermore, skin conductance responses increased when feedback was coupled with reinforcement, an effect which was larger in children with ADHD than controls. This could be interpreted as demonstrating that children with ADHD suffer from a diminished awareness of the significance of feedback in the feedback-only condition. The current study suggests that children with ADHD suffer from motivation problems when reinforcement was not available, at least when variability in responding was measured. Underestimations of time may reflect more stable deficits in ADHD.


'When coercion moves into your home'--a qualitative study of patient experiences with outpatient commitment in Norway.
The use of coercion on people with mental health problems is a serious intervention, and a reduction in its use is a declared goal in mental healthcare. Yet, many countries have introduced expanded powers of coercion in recent years, including outpatient commitment (OC). However, the evidence of the effectiveness of OC is inconclusive, and little is known about how patients experience OC schemes. The objective of this qualitative study was to explore (i) patients' experiences with OC, and (ii) how routines in care and health services affect patients' everyday living. The data were collected in 2011-2012 and included 11 qualitative in-depth interviews with patients subject to OC. The study used a narrative approach to interviews and a thematic narrative analysis. Participants generally complied with the OC requirements because of the clear and secure framework of OC, and also because they believed that the alternative would be involuntary hospitalisation. No one reported physical force, but coercion was experienced as limitation of freedom of action through excessive control and little patient influence or participation in their own treatment. Factors affecting patients' freedom of action under OC should be taken into account when the imposition of an OC order is considered.


Latest research progress in the correlation between baicalein and breast cancer invasion and metastasis.
Breast cancer is one of the most commonly occurring female malignant tumors. According to the 2012 GLOBOCAN statistics, produced by the International Agency for Research On Cancer ('IARC'), nearly 1.7 million women were diagnosed with breast cancer, with 522,000 related deaths: An increase in the incidence of breast cancer and associated mortality by nearly 18% from 2008. Metastasis is the final step in breast cancer progression, and represents the most common cause of mortality in patients with breast cancer. Therefore, a search for low-toxicity, safe and effective anti-breast cancer drugs in the form of natural compounds has become an intense focus of research. Baicalein, a widely used Chinese herbal medicine, has extensive antitumor activity. The present review briefly describes the research that has been performed on the association between baicalein and breast cancer metastasis, and further illustrates the influence of baicalein on the underlying mechanisms of breast cancer metastasis, adding a novel theory basis for baicalein antitumor research. In conclusion, baicalein may represent a promising target for the prevention and therapy of breast cancer.


Safety and immunogenicity of influenza vaccine among HIV-infected adults: Conventional vaccine vs. intradermal vaccine.
Several studies have reported poor immune responses to conventional influenza vaccines in HIV-infected individuals. This study sought to elicit more potent immunogenicity in HIV-infected adults using an intradermal vaccine compared with a conventional intramuscular vaccine. This multicenter, randomized, controlled, open-label study was conducted at 3 university hospitals during the 2011/2012 pre-influenza season. Three vaccines were used in HIV-infected adults aged 18 - 60 years: an inactivated intramuscular vaccine (Agrippal), a reduced-content intradermal vaccine (IDflu9μg) and a standard-content intradermal vaccine (IDflu15μg). Serum hemagglutination-inhibiting (HI) antibodies and INF-γ ELISpot assay were measured at the time of vaccination and 1 month after vaccination. Adverse events were recorded for 7 d. A total of 28 Agrippal, 30 IDflu9μg, and 28 IDflu15μg volunteers were included in this analysis. One month after vaccination, the GMTs and differences in INF-γ ELISpot assay results were similar among the 3 groups. Seroprotection rates, seroconversion rates and mean fold increases (MFI) among the 3 groups were also similar, at approximately 80%, 50-60% and 2.5 - 10.0, respectively. All three vaccines satisfied the CHMP criteria for the A/H1N1 and A/H3N2 strains, but not those for the B strain. In univariate analysis, no demographic or clinical factors, including age, CD4+ T-cell counts, HIV viral load, ART status and vaccine type, were related to failure to achieve seroprotection. The three vaccines were all well-tolerated and all reported reactions were mild to moderate. However, there was a tendency toward a higher incidence of local and systemic reactions in the intradermal vaccine groups. The intradermal vaccine did not result in higher immunogenicity compared to the conventional intramuscular vaccine, even with increased antigen dose.


Assessing Prognosis of Areas of Acute Cerebral Ischemia Using Perfusion CT.
Vascular recanalization by the vascular reconstruction method can dramatically improve ischemic symptoms in patients with acute cerebral ischemia. However, this treatment method is frequently associated with haemorrhagic complications. The indications for this therapeutic approach have been described in a number of studies in the literature. The present paper discusses the possibility of assessing the prognosis of ischemic areas using perfusion CT (PCT) by comparing the results obtained before and after thrombolytic therapy. Twenty-six patients underwent vascular reconstruction at our hospital between July 2002 and March 2004. Of these patients, six who underwent PCT before treatment and showed adequate recanalization following vascular reconstruction were included in the present study. PCT images were obtained using the first-pass bolus-tracking method with a 16-row multislice helical CT scanner. Areas of cerebral ischemia were evaluated by CT before and after vascular reconstruction. A region of interest was placed in the area showing low density in CT images before vascular reconstruction. The mean average CBF (mL/min/100 g), CBV (mL/100 g), and MTT (s) values were calculated in areas with and without cerebral infarction after vascular reconstruction. The %CBF, % CBV, and %MTT values relative to the normal side were evaluated with reference to the time until recanalization. Transarterial vascular reconstruction resulted in full recanalization in four patients and partial recanalization in two. The mean time from onset to recanalization was 284.7 +/- 63.27 minutes and was not longer than six hours in any patient. The patient prognosis results in terms of GOS were GR in two patients, MD in three patients, and SD in one patient. Based on comparison of the time after examination to recanalization, the %CBF showed a significant positive correlation in the salvaged area (Y = 47.321 + 2.491 x %CBF:R(2) = 0.792, p < 0.05). A significant correlation was not observed in %CBV, but %MTT showed a significant negative correlation (Y = 269.45 - 0.356 x %MTT:R(2) = 0.794, p < 0.05). The %CBF and %MTT results obtained by PCT performed before transarterial vascular reconstruction suggest that it may be possible to estimate the time before vascular reconstruction and the relationship with prognosis. These findings are expected to help ensure the appropriate application of vascular reconstruction and to provide useful information for developing optimal therapeutic protocols, thus reducing complications. In addition, because the results are based on the time after examination, the appropriate therapeutic approach can be determined even when the time of onset of ischemia is uncertain.


Current controversies in pediatric transplantation.
Transplantation in children has become a therapeutic option for several end stage organ diseases. The kidney, liver, and heart are the most common organs' transplanted; however, an increasing number of children are undergoing successful intestine, lung, and multiple organ transplant combinations. Through case study reports, emerging transplant options for the child experiencing end stage liver, intestine, heart, and lung failure are described. Critical care nurses play a crucial role in the postoperative recovery of these patients. An understanding of the transplant process and consequences of immunosuppression will help the critical care nurse identify signs of rejection, infection, and posttransplant lymphoproliferative disease.


Approaches for evaluating veterinary epidemiological models: verification, validation and limitations.
The evaluation of models of the spread and control of animal diseases is crucial if these models are to be used to inform decisions about the control or management of such diseases. Two key steps in the evaluation of epidemiological models are model verification and model validation. Verification is the demonstration that a computer-driven model is operating correctly, and conforms to its intended design. Validation refers to the process of determining how well a model corresponds to the system that it is intended to represent. For a veterinary epidemiological model, validation would address such issues as how well the model represents the dynamics of the disease in question in the population to which this model is applied, and how well the model represents the application of different measures for disease control. Just as the development of epidemiological models is a subjective, continuous process, subject to change and refinement, so too is the evaluation of models. The purpose of model evaluation is not to demonstrate that a model is a 'true' or accurate' representation of a system, but to subject it to sufficient scrutiny so that it may be used with an appropriate degree of confidence to aid decision-making. To facilitate model verification and validation, epidemiological modellers should clearly state the purpose, assumptions and limitations of a model; provide a detailed description of the conceptual model; document those steps already taken to test the model; and thoroughly describe the data sources and the process used to produce model input parameters from those data.


Religious and national group identification in adolescence: a study among three religious groups in Mauritius.
Religious group identification is an important but understudied social identity. The present study investigates religious group identification among adolescents of different faiths (Hindu, Muslim, Christian) living in multicultural Mauritius. It further explores how religious and national group identities come together among religious majority and minority adolescents. For three age groups (11 to 19 years, N = 2152) we examined the strength of adolescents' religious and national group identification, the associations between these two identities, and the relationships to global self-esteem. Across age and religious group, participants reported stronger identification with their religious group than with the nation. Identification with both categories declined with age, with the exception of Muslims, whose strong religious identification was found across adolescence. The association between religious and national identification was positive, albeit stronger for the majority group of Hindus and for early adolescents. We examined the manner in which religious and national identities come together using a direct self-identification measure and by combining the separate continuous measures of identification. Four distinct clusters of identification (predominant religious identifiers, dual identifiers, neutrals, and separate individuals) that were differently associated with global self-esteem were found. Dual identifiers reported the highest level of global self-esteem. The clusters of identification did not fully correspond to the findings for the direct self-identification measure. The results are discussed in terms of the meaning of dual identity and the positive manner in which adolescents can manage their multiple identities while taking into account the ideological framework in which those identities are played out.


The concurrent maturation of mouse and human thymocytes in human fetal thymus implanted in NIH-beige-nude-xid mice is associated with the reconstitution of the murine immune system.
To determine whether the human thymus provides an environment for the maturation of murine T cells, human fetal thymus and liver (hu-thy/liv) were implanted into congenitally athymic NIH-beige-nude-xid (BNX) mice or C.B-17 scid/scid (SCID) mice. 3 mo after implantation, in contrast to the hu-thy/liv implant in SCID mice, which was populated only with human CD4/CD8 single- and double-positive thymocytes, the hu-thy/liv implant in BNX mice contained a chimeric population of human and mouse CD4/CD8 single- and double-positive thymocytes. Immunohistochemical staining of the hu-thy/liv implant in BNX mice indicated that the population of double-positive mouse thymocytes was localized to discrete areas of the human fetal thymus. Quantitative improvements in mouse T cell and immunoglobulin (Ig) G parameters were observed after grafting of the human fetal thymus and liver tissue into BNX mice. In addition, in contrast to the nonimplanted BNX mice, the implanted BNX mice were capable of mounting a keyhole limpet hemocyanin-specific IgG response and their peripheral T cells were responsive to stimulation with mitogens and antibodies directed to the T cell receptor. Furthermore, after in vivo priming, T cells present in lymph nodes of the implanted BNX mice were capable of mounting an antigen-induced in vitro T cell-dependent proliferative response. Thus, concurrent with the continued maturation of human T cells, murine T cells differentiated within the human fetal thymus implanted in the BNX mice and mediated the phenotypic and functional reconstitution of the murine immune system. Mice with a reconstituted immune system that contain a human thymic implant that is infectible with human immunodeficiency virus (HIV) should prove useful in the investigation of T cell maturation in the thymus and in the evaluation of potential HIV vaccines.


Increased severity of the CHIMERA model induces acute vascular injury, sub-acute deficits in memory recall, and chronic white matter gliosis.
Traumatic brain injury (TBI) is a leading cause of death and disability in modern societies. Diffuse axonal and vascular injury are nearly universal consequences of mechanical energy impacting the head and contribute to disability throughout the injury severity spectrum. CHIMERA (Closed Head Impact Model of Engineered Rotational Acceleration) is a non-surgical, impact-acceleration model of rodent TBI that reliably produces diffuse axonal injury characterized by white matter gliosis and axonal damage. At impact energies up to 0.7 joules, which result in mild TBI in mice, CHIMERA does not produce detectable vascular or grey matter injury. This study was designed to expand CHIMERA's capacity to induce more severe injuries, including vascular damage and grey matter gliosis. This was made possible by designing a physical interface positioned between the piston and animal's head to allow higher impact energies to be transmitted to the head without causing skull fracture. Here, we assessed interface-assisted single CHIMERA TBI at 2.5 joules in wild-type mice using a study design that spanned 6 h-60 d time points. Injured animals displayed robust acute neurological deficits, elevated plasma total tau and neurofilament-light levels, transiently increased proinflammatory cytokines in brain tissue, blood-brain barrier (BBB) leakage and microstructural vascular abnormalities, and grey matter microgliosis. Memory deficits were evident at 30 d and resolved by 60 d. Intriguingly, white matter injury was not remarkable at acute time points but evolved over time, with white matter gliosis being most extensive at 60 d. Interface-assisted CHIMERA thus enables experimental modeling of distinct endophenotypes of TBI that include acute vascular and grey matter injury in addition to chronic evolution of white matter damage, similar to the natural history of human TBI.


Non-organic failure to thrive: a reappraisal.
Non-organic failure to thrive has traditionally been regarded as due primarily to maternal rejection and neglect. A critical reappraisal of the evidence suggests a more balanced view of the mother-child relationship should be taken. A classification of the condition, founded on facts not concepts, is urgently required. Non-organic failure to thrive should be viewed in a multidimensional context, in which potential influences upon the symptomatic infant are considered. Inadequacy of nutrition is caused by both a failure of adequate provision of food and by inadequate intake. A vicious circle of maladaptive behavioural interaction between caregiver and infant is often present, sustained by high emotional tensions. Clinical intervention should aim to clarify the contributions made by both caregiver and infant to that interaction and thus break the cycle. The basis on which intervention is made should be direct observation of the parent and child relationship in as many different environmental contexts as feasible, especially during feeding. The multidisciplinary team has an important role to play in management. An emphasis on parental culpability in the aetiology of non-organic failure to thrive, in the absence of direct evidence of neglect, is wrong.


Biological monitoring of exposure to benzene: a comparison between S-phenylmercapturic acid, trans,trans-muconic acid, and phenol.
Comparison of the suitability of two minor urinary metabolites of benzene, trans,trans-muconic acid (tt-MA) and S-phenylmercapturic acid (S-PMA), as biomarkers for low levels of benzene exposure. The sensitivity of analytical methods of measuring tt-MA and S-PMA were improved and applied to 434 urine samples collected from 188 workers in 12 studies in different petrochemical industries and from 52 control workers with no occupational exposure to benzene. In nine studies airborne benzene concentrations were assessed by personal air monitoring. Strong correlations were found between tt-MA and S-PMA concentrations in samples from the end of the shift and between either of these variables and airborne benzene concentrations. It was calculated that exposure to 1 ppm (8 hour time weighted average (TWA)) benzene leads to an average concentration of 1.7 mg tt-MA and 47 micrograms S-PMA/g creatinine in samples from the end of the shift. It was estimated that, on average, 3.9% (range 1.9%-7.3%) of an inhaled dose of benzene was excreted as tt-MA with an apparent elimination half life of 5.0 (SD 2.3) hours and 0.11% (range 0.05%-0.26%) as S-PMA with a half life of 9.1 (SD 3.7) hours. The mean urinary S-PMA in 14 moderate smokers and 38 non-smokers was 3.61 and 1.99 micrograms/g creatinine, respectively and the mean urinary tt-MA was 0.058 and 0.037 mg/g creatinine, respectively. S-PMA proved to be more specific and more sensitive (P = 0.030, Fisher's exact test) than tt-MA. S-PMA, but not tt-MA, was always detectable in the urine of smokers who were not occupationally exposed. S-PMA was also detectable in 20 of the 38 non-smokers from the control group whereas tt-MA was detectable in only nine of these samples. The inferior specificity of tt-MA is due to relatively high background values (up to 0.71 mg/g creatinine in this study) that may be found in non-occupationally exposed people. Although both tt-MA and S-PMA are sensitive biomarkers, only S-PMA allows reliable determination of benzene exposures down to 0.3 ppm (8 h TWA) due to its superior specificity. Because it has a longer elimination half life S-PMA is also a more reliable biomarker than tt-MA for benzene exposures during 12 hour shifts. For biological monitoring of exposure to benzene concentrations higher than 1 ppm (8 h TWA) tt-MA is also suitable and may even be preferred due to its greater ease of measurement.


Impact of Organic Carbon Electron Donors on Microbial Community Development under Iron- and Sulfate-Reducing Conditions.
Although iron- and sulfate-reducing bacteria in subsurface environments have crucial roles in biogeochemical cycling of C, Fe, and S, how specific electron donors impact the compositional structure and activity of native iron- and/or sulfate-reducing communities is largely unknown. To understand this better, we created bicarbonate-buffered batch systems in duplicate with three different electron donors (acetate, lactate, or glucose) paired with ferrihydrite and sulfate as the electron acceptors and inoculated them with subsurface sediment as the microbial inoculum. Sulfate and ferrihydrite reduction occurred simultaneously and were faster with lactate than with acetate. 16S rRNA-based sequence analysis of the communities over time revealed that Desulfotomaculum was the major driver for sulfate reduction coupled with propionate oxidation in lactate-amended incubations. The reduction of sulfate resulted in sulfide production and subsequent abiotic reduction of ferrihydrite. In contrast, glucose promoted faster reduction of ferrihydrite, but without reduction of sulfate. Interestingly, the glucose-amended incubations led to two different biogeochemical trajectories among replicate bottles that resulted in distinct coloration (white and brown). The two outcomes in geochemical evolution might be due to the stochastic evolution of the microbial communities or subtle differences in the initial composition of the fermenting microbial community and its development via the use of different glucose fermentation pathways available within the community. Synchrotron-based x-ray analysis indicated that siderite and amorphous Fe(II) were formed in the replicate bottles with glucose, while ferrous sulfide and vivianite were formed with lactate or acetate. These data sets reveal that use of different C utilization pathways projects significant changes in microbial community composition over time that uniquely impact both the geochemistry and mineralogy of subsurface environments.


Lipid peroxidation induced by Clinostomum detruncatum in muscle of the freshwater fish Rhamdia quelen.
The effect of Clinostomum detruncatum metacercaria infection on the activities of the antioxidant enzymes superoxide dismutase and catalase in muscle of the freshwater fish Rhamdia quelen was analyzed. Tert-butyl hydroperoxide-initiated chemiluminescence, a measure of lipid peroxidation, was also investigated. Enzyme activities were similar in infected and uninfected fishes. However, the chemiluminescence was almost 2-fold higher in muscle of infected fishes than in muscle of uninfected ones. These results indicate that parasite infection induces oxidative stress and a higher level of membrane damage in the fish muscle due to an imbalance between pro-oxidants and non-enzymatic antioxidants. Our results suggest that fish response to parasite infection could involve, as in other vertebrates, reactive oxygen intermediates.


Rate and time to develop first central line-associated bloodstream infections when comparing open and closed infusion containers in a Brazilian Hospital.
The objective of the study was to determine the effect of switching from an open (glass or semi-rigid plastic) infusion container to a closed, fully collapsible plastic infusion container (Viaflex) on rate and time to onset of central lineassociated bloodstream infections (CLABSI). An open-label, prospective cohort, active healthcare-associated infection surveillance, sequential study was conducted in three intensive care units in Brazil. The CLABSI rate using open infusion containers was compared to the rate using a closed infusion container. Probability of acquiring CLABSI was assessed over time and compared between open and closed infusion container periods; three-day intervals were examined. A total of 1125 adult ICU patients were enrolled. CLABSI rate was significantly higher during the open compared with the closed infusion container period (6.5 versus 3.2 CLABSI/1000 CL days; RR=0.49, 95%CI=0.26- 0.95, p=0.031). During the closed infusion container period, the probability of acquiring a CLABSI remained relatively constant along the time of central line use (0.8% Days 2-4 to 0.7% Days 11-13) but increased in the open infusion container period (1.5% Days 2-4 to 2.3% Days 11-13). Combined across all time intervals, the chance of a patient acquiring a CLABSI was significantly lower (55%) in the closed infusion container period (Cox proportional hazard ratio 0.45, p= 0.019). CLABSIs can be reduced with the use of full barrier precautions, education, and performance feedback. Our results show that switching from an open to a closed infusion container may further reduce CLABSI rate as well as delay the onset of CLABSIs. Closed infusion containers significantly reduced CLABSI rate and the probability of acquiring CLABSI.


Eicosapentaenoic acid-induced endothelium-dependent and -independent relaxation of sheep pulmonary artery.
It is known that long chain polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA), have beneficial effects on cardiovascular function including pulmonary hypertension. The purpose of the present study was to examine the mechanisms involved in EPA-induced relaxation of sheep isolated pulmonary artery by measuring isometric tension. Nitric oxide (NO) derived from constitutive nitric oxide synthase (cNOS) was measured by Greiss method in the presence of the inducible nitric oxide synthase (iNOS) selective inhibitor N-[[3-(aminomethyl) phenyl]methyl]-ethanimidamide, dihydrochloride (1400 W). EPA (10(-)(7)-10(-)(4)M) caused concentration-dependent relaxation of sheep pulmonary artery with a pD(2) of 5.56+/-0.09 and E(max) of 87.40+/-3.10% (n=9). N(G)-nitro-L-arginine methyl ester (L-NAME) 100 microM significantly attenuated (E(max) 41.95+/-6.70%; n=8) EPA-induced relaxation of endothelium intact arterial rings. Similarly, endothelium denudation markedly inhibited (E(max) 17.60+/-1.21%; n=4) EPA-induced relaxation. EPA (30 microM) significantly increased the cNOS-derived NO release (10.17+/-0.96; n=8 versus control 7.43+/-0.78 pmol/mg tissue wet wt./h; n=7) in endothelium intact vessels. However, EPA-stimulated NO release was markedly blunted by either 100 microM L-NAME (7.07+/-0.54 pmol/mg tissue wet wt./h; n=8) or endothelium removal (6.97+/-0.87 pmol/mg tissue wet wt./h; n=17). In endothelium-denuded K(+) (80 mM)-depolarized arterial rings, EPA (30 microM) significantly inhibited CaCl(2)-induced contractions (E(max) 42.77+/-5.90% versus control 94.78+/-9.82%; n=5). The fatty acid also inhibited nifedipine (1 microM)-insensitive 5-HT-induced contractions in this vessel (E(max) 70.57+/-4.88% versus control 161.50+/-17.46%; n=5). In conclusion, EPA relaxes sheep pulmonary artery primarily through endothelium-dependent NO release, and the residual endothelium-independent relaxation may result from inhibition of Ca(2+)-influx through L-type calcium channels, as well as 5-HT-stimulated intracellular Ca(2+) release.


Numerical study on an application of subwavelength dielectric gratings for high-sensitivity plasmonic detection.
Although subwavelength dielectric gratings can be employed to achieve a high sensitivity of the surface plasmon resonance (SPR) biosensor, the plasmonic interpretation verifying the resulting sensitivity improvement remains unclear. The aim of this study is to elucidate the effects of the grating's geometric parameters on the amplification of SPR responses and to understand the physical mechanisms associated with the enhancement. Our numerical results show that the proposed SPR substrate with a dielectric grating can provide a better sensitivity due to the combined effects of surface reaction area and field distribution at the binding region. An influence of adhesion layer on the sensor performance is also discussed. The obtained results will be promising in high-sensitivity plasmonic biosensing applications.


High cholera vaccination coverage following emergency campaign in Haiti: Results from a cluster survey in three rural Communes in the South Department, 2017.
Oral cholera vaccine (OCV) has increasingly been used as an outbreak control measure, but vaccine shortages limit its application. A two-dose OCV campaign targeting residents aged over 1 year was launched in three rural Communes of Southern Haiti during an outbreak following Hurricane Matthew in October 2016. Door-to-door and fixed-site strategies were employed and mobile teams delivered vaccines to hard-to-reach communities. This was the first campaign to use the recently pre-qualified OCV, Euvichol. The study objective was to estimate post-campaign vaccination coverage in order to evaluate the campaign and guide future outbreak control strategies. We conducted a cluster survey with sampling based on random GPS points. We identified clusters of five households and included all members eligible for vaccination. Local residents collected data through face-to-face interviews. Coverage was estimated, accounting for the clustered sampling, and 95% confidence intervals calculated. 435 clusters, 2,100 households and 9,086 people were included (99% response rate). Across the three communes respectively, coverage by recall was: 80.7% (95% CI:76.8-84.1), 82.6% (78.1-86.4), and 82.3% (79.0-85.2) for two doses and 94.2% (90.8-96.4), 91.8% (87-94.9), and 93.8% (90.8-95.9) for at least one dose. Coverage varied by less than 9% across age groups and was similar among males and females. Participants obtained vaccines from door-to-door vaccinators (53%) and fixed sites (47%). Most participants heard about the campaign through community 'criers' (58%). Despite hard-to-reach communities, high coverage was achieved in all areas through combining different vaccine delivery strategies and extensive community mobilisation. Emergency OCV campaigns are a viable option for outbreak control and where possible multiple strategies should be used in combination. Euvichol will help alleviate the OCV shortage but effectiveness studies in outbreaks should be done.


New insights into the enzymatic mechanism of human chitotriosidase (CHIT1) catalytic domain by atomic resolution X-ray diffraction and hybrid QM/MM.
Chitotriosidase (CHIT1) is a human chitinase belonging to the highly conserved glycosyl hydrolase family 18 (GH18). GH18 enzymes hydrolyze chitin, an N-acetylglucosamine polymer synthesized by lower organisms for structural purposes. Recently, CHIT1 has attracted attention owing to its upregulation in immune-system disorders and as a marker of Gaucher disease. The 39 kDa catalytic domain shows a conserved cluster of three acidic residues, Glu140, Asp138 and Asp136, involved in the hydrolysis reaction. Under an excess concentration of substrate, CHIT1 and other homologues perform an additional activity, transglycosylation. To understand the catalytic mechanism of GH18 chitinases and the dual enzymatic activity, the structure and mechanism of CHIT1 were analyzed in detail. The resolution of the crystals of the catalytic domain was improved from 1.65 Å (PDB entry 1waw) to 0.95-1.10 Å for the apo and pseudo-apo forms and the complex with chitobiose, allowing the determination of the protonation states within the active site. This information was extended by hybrid quantum mechanics/molecular mechanics (QM/MM) calculations. The results suggest a new mechanism involving changes in the conformation and protonation state of the catalytic triad, as well as a new role for Tyr27, providing new insights into the hydrolysis and transglycosylation activities.


Does cervical radiculopathy have an effect on peripheral nerve conduction studies? An electrophysiological evaluation.
Peripheral nerve neuropathies are more common in patients with cervical radiculopathy (CR) and a proximal lesion along an axon might predispose that nerve to injury at distal sites. To evaluate this hypothesis, the frequency of median nerve neuropathy at the wrist and the frequency of ulnar nerve neuropathy at the elbow were investigated in 80 patients with one-sided CR. The symptomatic and asymptomatic extremities were compared for the electrophysiological parameters of ulnar and median nerve conduction studies. The patients were divided into three diagnostic subgroups (C5/C6 radiculopathy, C7 radiculopathy and C8/Th1 radiculopathy), and ulnar nerve conduction studies were compared between symptomatic and asymptomatic extremities in each subgroup. Although, the median and ulnar nerves are mainly derived from C8/Th1 roots, we did not observe an increased frequency of ulnar nerve involvement at the elbow (3.7% in symptomatic, 2.5% in asymptomatic extremities) while increased median nerve involvement was present at the wrist (27.5% in symptomatic, 12.5% in asymptomatic extremities). The electrophysiological data and the frequency of ulnar and median nerve neuropathy did not support an etiological association between CR and distal neuropathy. the association between CR and peripheral neuropathy is associational rather than causative.


The risk of coronary thrombosis with cyclo-oxygenase-2 inhibitors does not vary with polymorphisms in two regions of the cyclo-oxygenase-2 gene.
To investigate whether polymorphisms of the cyclo-oxygenase-2 (COX-2) gene modify the adverse cardiovascular effects of COX-2 inhibitors. A case control study was conducted in the Hunter Region of New South Wales, Australia. Cases (n= 460) were hospitalized with acute coronary syndrome (ACS). Controls (n= 640) were recruited from the electoral rolls. Structured interviews gathered information on variables including recent ingestion of non-steroidal anti-inflammatory drugs (NSAIDs). Targeted genotyping of rs 20417(G > C) and rs5275 (T > C) polymorphisms was performed by real-time polymerase chain reaction using allele-specific probes. Ingestion of any NSAID in the week prior to interview was associated with an elevated risk for ACS: adjusted odds ratio 1.8 (1.2, 2.5). The rs 20417 and rs 5275 polymorphisms were not singly associated with risk for ACS: adjusted odds ratios 1.1 (0.80, 1.5) and 1.2 (0.88, 1.5), respectively. Individually, the polymorphisms did not modify the risk of ACS with the drugs. When analyses were conducted by haplotype, the adjusted odds ratio with celecoxib or rofecoxib in individuals who had one or two copies of the 'low risk' haplotype (no GT) was 1.2 (0.29, 5.0), compared with 2.1 (1.1, 4.0) with the 'high risk' haplotype (one or two copies of GT). We found little evidence of a gene/drug interaction. We found a statistically non-significant trend toward a lower risk of coronary events with NSAIDs in the presence of the 'low risk' haplotype. Even if confirmed, the clinical utility of the finding would be limited as this haplotype is carried by a minority of the population.


Effects of a high alpha-linolenate and high linoleate diet on hemolysis and lipid peroxidation of rat erythrocytes.
alpha-Linolenic acid (18:3n-3) is known to autoxidize several fold faster than linoleic acid (18:2n-6). Feeding a high alpha-linolenate or a high linoleate diet to rats resulted in significant changes in the n-3/n-6 ratios of 20 and 22 carbon highly unsaturated fatty acids in erythrocytes. However, the rates of hemolysis observed in N2- or O2-atmosphere were similar between the two dietary groups. No significant amounts of conjugated dienes were detected and no measurable changes in the fatty acid compositions were observed during the incubations, indicating that the hemolysis occurred without involving significant lipid peroxidation. When stimulated with a free radical initiator, [2,2'-azobis-(2-amidinopropane)dihydrochloride] (AAPH), hemolysis occurred more rapidly, conjugated dienes formed and unsaturated/saturated ratios of phospholipid fatty acids decreased. However, no statistically significant difference was observed in these parameters of the two dietary groups. These results indicate that hemolysis occurs without involving lipid peroxidation but is accelerated by free radicals through lipid peroxidation, and that the difference in autoxidizabilities of alpha-linolenate and linoleate is not reflected in the rates of hemolysis and autoxidation in rat erythrocytes.


Immune activation and neuropsychiatric symptoms in HIV infection.
This study examined the role of biological processes in the development of specific neuropsychiatric complications in HAART-naive adults with HIV/AIDS. Depressive symptoms were modestly associated with elevated IL-6 mRNA expression (r(s)=0.40, p<0.05) even after removing the influences of other subjective complaints (pr=0.39, p<0.05). Elevated serum neopterin was strongly associated with depressive symptoms in individuals taking antidepressants (r(s)=0.83, p<0.001), though the association was nullified in those not on antidepressants (r(s)=-0.25, p>0.05). Mean neopterin levels were higher in the depressed as compared with nondepressed group but only for those taking antidepressants (F=45.66, df=1, 11, p<0.001). Neuropsychological impairment was not associated with the biological markers. These findings suggest that systemic immune markers (like neopterin) may be useful in differentiating treatment-resistant individuals at greater risk of developing chronic depression.


Evaluation of substantivity of hypochlorous acid as an antiplaque agent: A randomized controlled trial.
Hypochlorous acid (HOCl) is a non-antibiotic antimicrobial substance with significant effects on pathogenic oral micro-organisms. The effects of HOCl as an antiplaque agent have not been studied. The aim of this study was to evaluate the substantivity of HOCl mouthwashes compared with chlorhexidine (CHX) rinses and a placebo. A double-blind randomized controlled trial with 75 participants was conducted. Participants were divided into five groups using block randomization: 0.025% HOCl, 0.05% HOCl, 0.12% CHX, 0.2% CHX, and sterile water as a placebo. Participants were instructed to use each rinse solution for 30 seconds after dental prophylaxis. Samples of saliva were taken at baseline and after 30 seconds, 1, 3, 5 and 7 hours to assess substantivity, and bacterial viability was established by the fluorescence method. Visible plaque in all participants was assessed with the Turesky index at baseline and at 7 hours, and adverse events were also assessed. HOCl led to a 33% reduction in bacterial counts in the saliva after 30 seconds compared with a 58% reduction by CHX. HOCl has no substantivity, and bacterial counts returned to baseline after 1 hour. Placebo treatment led to the highest plaque count after 7 hours compared with the CHX and HOCl groups, although the differences were not significant. HOCl rinsing induced the highest percentages of unpleasant taste and dryness sensations. HOCl rinses have an initial effect on bacterial viability in saliva but have no substantivity. Other mechanisms may explain its antiplaque effect.


Anti-Inflammatory Effects of Cannabigerol in Rheumatoid Arthritis Synovial Fibroblasts and Peripheral Blood Mononuclear Cell Cultures Are Partly Mediated by TRPA1.
Since its medical legalization, cannabis preparations containing the major phytocannabinoids (cannabidiol (CBD) and δ9-tetrahydrocannabinol (THC)) have been used by patients with rheumatoid arthritis (RA) to alleviate pain and inflammation. However, minor cannabinoids such as cannabigerol (CBG) also demonstrated anti-inflammatory properties, but due to the lack of studies, they are not widely used. CBG binds several cellular target proteins such as cannabinoid and α2-adrenergic receptors, but it also ligates several members of the transient potential receptor (TRP) family with TRPA1 being the main target. TRPA1 is not only involved in nnociception, but it also protects cells from apoptosis under oxidative stress conditions. Therefore, modulation of TRPA1 signaling by CBG might be used to modulate disease activity in RA as this autoimmune disease is accompanied by oxidative stress and subsequent activation of pro-inflammatory pathways. Rheumatoid synovial fibroblasts (RASF) were stimulated or not with tumor necrosis factor (TNF) for 72 h to induce TRPA1 protein. CBG increased intracellular calcium levels in TNF-stimulated RASF but not unstimulated RASF in a TRPA1-dependent manner. In addition, PoPo3 uptake, a surrogate marker for drug uptake, was enhanced by CBG. RASF cell viability, IL-6 and IL-8 production were decreased by CBG. In peripheral blood mononuclear cell cultures (PBMC) alone or together with RASF, CBG-modulated interleukin (IL)-6, IL-10, TNF and immunoglobulin M and G production which was dependent on activation stimulus (T cell-dependent or independent). However, effects on PBMCs were only partially mediated by TRPA1 as the antagonist A967079 did inhibit some but not all effects of CBG on cytokine production. In contrast, TRPA1 antagonism even enhanced the inhibitory effects of CBG on immunoglobulin production. CBG showed broad anti-inflammatory effects in isolated RASF, PBMC and PBMC/RASF co-cultures. As CBG is non-psychotropic, it might be used as add-on therapy in RA to reduce IL-6 and autoantibody levels.


Interactions between somatic cells and germ cells throughout mammalian oogenesis.
Oocytes and their companion somatic cells maintain a close association throughout oogenesis and this association is essential for normal oocyte and follicular development. This review summarizes current concepts of the role of the somatic cells in the regulation of mammalian oocyte growth, the maintenance of meiotic arrest, the induction of oocyte maturation, and the acquisition of full embryonic developmental competence during oocyte maturation in vitro. Gap junctions appear to mediate these regulatory processes. The regulatory interaction of oocytes and somatic cells, however, is not unidirectional; the oocyte participates in the proliferation, development, and function of the follicular somatic cells. The oocyte secretes factors that enable the cumulus cells to synthesize hyaluronic acid and undergo cumulus expansion in response to hormonal stimulation. In addition, the oocyte produces factors that promote the proliferation of granulosa cells. These interactions in vitro do not appear to require the mediation of gap junctions. The oocyte also promotes the differentiation of granulosa cells into functional cumulus cells, but this function of the oocyte appears to require the continued presence and close association of the oocyte and granulosa cells. Therefore, oocytes and follicular somatic cells are interdependent for development and function.