Mechanisms of IFN-γ-induced apoptosis of human skin keratinocytes in patients with atopic dermatitis.
Enhanced apoptosis of keratinocytes is the main cause of eczema and spongiosis in patients with the common inflammatory skin disease atopic dermatitis (AD). The aim of the study was to investigate molecular mechanisms of AD-related apoptosis of keratinocytes. Primary keratinocytes isolated from patients with AD and healthy donors were used to study apoptosis by using annexin V/7-aminoactinomycin D staining. Illumina mRNA Expression BeadChips, quantitative RT-PCR, and immunofluorescence were used to study gene expression. In silico analysis of candidate genes was performed on genome-wide single nucleotide polymorphism data. We demonstrate that keratinocytes of patients with AD exhibit increased IFN-γ-induced apoptosis compared with keratinocytes from healthy subjects. Further mRNA expression analyses revealed differential expression of apoptosis-related genes in AD keratinocytes and skin and the upregulation of immune system-related genes in skin biopsy specimens of chronic AD lesions. Three apoptosis-related genes (NOD2, DUSP1, and ADM) and 8 genes overexpressed in AD skin lesions (CCDC109B, CCL5, CCL8, IFI35, LYN, RAB31, IFITM1, and IFITM2) were induced by IFN-γ in primary keratinocytes. The protein expression of IFITM1, CCL5, and CCL8 was verified in AD skin. In line with the functional studies and AD-related mRNA expression changes, in silico analysis of genome-wide single nucleotide polymorphism data revealed evidence of an association between AD and genetic markers close to or within the IFITM cluster or RAB31, DUSP1, and ADM genes. Our results demonstrate increased IFN-γ responses in skin of patients with AD and suggest involvement of multiple new apoptosis- and inflammation-related factors in the development of AD.


Update version of the Japanese Guidelines for Gastric Cancer Screening.
Although the incidence and mortality of gastric cancer have gradually decreased, its burden remains in East Asian countries. Gastric cancer screening has been performed in Japan since 1983, and the introduction of new screening techniques has been eagerly anticipated. To promote evidence-based screening, the Japanese guidelines for gastric cancer screening have been revised based on the new studies. The guidelines for gastric cancer screening have been developed according to a previously established method. To assess evidence regarding the effectiveness of the screening methods, a systematic review was conducted based on an analytic framework including clinical questions aiming at reducing mortality from gastric cancer. The following methods were assessed for gastric cancer screening: upper gastrointestinal series (radiographic screening), gastrointestinal endoscopy (endoscopic screening), Helicobacter pylori antibody test and serum pepsinogen tests. Based on the balance of the benefits and harms of each screening method, recommendations for population-based and opportunistic screenings were formulated. After the Japanese guidelines for gastric cancer screening were published in 2005, several observational studies on radiographic and endoscopic screenings have been reported. Three case-control studies have evaluated mortality reduction from gastric cancer by endoscopic screening. Notably, evidence of the H. pylori antibody and serum pepsinogen tests was insufficient. Although false-positive results, false-negative results, and complications were observed in endoscopic and radiographic screenings, the complication rates were higher in endoscopic screening than in radiographic screening. Overdiagnosis was not estimated directly in both methods. Radiographic and endoscopic screenings for gastric cancer are recommended for population-based and opportunistic screenings. The H. pylori antibody and serum pepsinogen tests are not recommended for population-based screening because of insufficient evidence.


Magnitude of Missed Esophageal Adenocarcinoma After Barrett's Esophagus Diagnosis: A Systematic Review and Meta-analysis.
A proportion of patients with Barrett's esophagus (BE) are diagnosed with esophageal adenocarcinoma (EAC) within 1 year of an endoscopic examination that produced negative findings. These cases of missed cancers have not been well studied, despite current surveillance strategies for BE. We performed a systematic review and meta-analysis to determine the magnitude of missed EAC in cohorts of patients with BE. We searched MEDLINE, EMBASE, and Web of Science from their inception to May 31, 2015 to identify cohort studies of adults with BE (baseline nondysplastic BE ± BE with low-grade dysplasia) and at least a 3-year follow-up period, providing data on missed and incident EACs (diagnosed within 1 year and diagnosed more than 1 year after the initial endoscopy in which BE was diagnosed, respectively). The main outcome measure was pooled proportion of missed and incident EACs (of all EACs detected after initial endoscopy) among BE cohorts, using a random effects model. In a meta-analysis of 24 studies reporting on 820 missed and incident EACs, 25.3% were classified as missed (95% confidence interval: 16.4%-36.8%) and 74.7% as incident EACs (95% CI: 63.2%-83.6%), although there was substantial heterogeneity among studies (I2 = 74%). When the analysis was restricted to nondysplastic BE cohorts (15 studies), 23.9% of EACs were classified as missed (95% confidence interval: 15.3%-35.4%; I2 = 0%). In a meta-analysis of 10 studies with follow-up periods of ≥5 years (a total of 239 EACs), 22.0% were classified as missed (95% confidence interval: 8.7%-45.5%), with substantial heterogeneity (I2 = 68%). Among adults with nondysplastic BE (or BE with low-grade dysplasia) at their index endoscopy and at least a 3-year follow-up period, 25% of EACs are diagnosed within 1 year after the index endoscopy. Additional resources should be allocated to detect missed EAC.


Finding the weakest link: exploring integrin-mediated mechanical molecular pathways.
From the extracellular matrix to the cytoskeleton, a network of molecular links connects cells to their environment. Molecules in this network transmit and detect mechanical forces, which subsequently determine cell behavior and fate. Here, we reconstruct the mechanical pathway followed by these forces. From matrix proteins to actin through integrins and adaptor proteins, we review how forces affect the lifetime of bonds and stretch or alter the conformation of proteins, and how these mechanical changes are converted into biochemical signals in mechanotransduction events. We evaluate which of the proteins in the network can participate in mechanotransduction and which are simply responsible for transmitting forces in a dynamic network. Besides their individual properties, we also analyze how the mechanical responses of a protein are determined by their serial connections from the matrix to actin, their parallel connections in integrin clusters and by the rate at which force is applied to them. All these define mechanical molecular pathways in cells, which are emerging as key regulators of cell function alongside better studied biochemical pathways.


Urinary Concentrations of Polycyclic Aromatic Hydrocarbon Metabolites in Maté Drinkers in Rio Grande do Sul, Brazil.
Background: Consumption of maté, an infusion of the herb Ilex paraguariensis (yerba maté), is associated with increased risk of esophageal squamous cell carcinoma (ESCC), but the carcinogenic mechanism is unclear. Commercial brands of yerba maté contain high levels of carcinogenic polycyclic aromatic hydrocarbons (PAHs), which are acquired during the traditional drying process. The purpose of this study was to characterize exposure to PAHs in maté drinkers over a wide range of maté consumption.Methods: We recruited 244 adults who answered a questionnaire and collected a fasting spot urine specimen. We quantified urinary concentrations of seven PAH metabolites and assessed associations between self-reported recent maté consumption and urinary PAH metabolites by multivariate regression.Results: Recent maté consumption showed a significant dose-response association with 6 of 7 PAH metabolites in unadjusted models (Ptrend < 0.05). After adjustment for creatinine and potential confounders, concentrations of 2-naphthol, 1-hydroxyphenanthrene, and the sum of 2- and 3-hydroxyphenanthrene remained significantly associated with recent maté intake. The sum of the urinary concentrations of the phenanthrene metabolites was similar or higher among maté drinkers who did not smoke than among smokers who did not drink matéConclusions: Urinary concentrations of PAH metabolites were significantly associated with self-reported amounts of recent maté intake, and drinking maté increased urinary concentrations of some PAH metabolites as much as smoking cigarettes.Impact: Drinking maté is a source of exposure to potentially carcinogenic PAHs, consistent with the hypothesis that the PAH content of maté may contribute to the increased risk of ESCC in maté drinkers. Cancer Epidemiol Biomarkers Prev; 27(3); 331-7. ©2017 AACR.


The treatment effects of flaxseed-derived secoisolariciresinol diglycoside and its metabolite enterolactone on benign prostatic hyperplasia involve the G protein-coupled estrogen receptor 1.
Secoisolariciresinol diglucoside (SDG), a lignan extracted from flaxseed, has been shown to suppress benign prostatic hyperplasia (BPH). However, little is known about the mechanistic basis for its anti-BPH activity. The present study showed that enterolactone (ENL), the mammalian metabolite of SDG, shared the similar binding site of G1 on a new type of membranous estrogen receptor, G-protein-coupled estrogen eceptor 1 (GPER), by docking simulations method. ENL and G1 (the specific agonist of GPER) inhibited the proliferation of human prostate stromal cell line WPMY-1 as shown by MTT assay and arrested cell cycle at the G0/G1 phase, which was displayed by propidium iodide staining following flow cytometer examination. Silencing GPER by short interfering RNA attenuated the inhibitory effect of ENL on WPMY-1 cells. The therapeutic potential of SDG in the treatment of BPH was confirmed in a testosterone propionate-induced BPH rat model. SDG significantly reduced the enlargement of the rat prostate and the number of papillary projections of prostatic alveolus and thickness of the pseudostratified epithelial and stromal cells when comparing with the model group. Mechanistic studies showed that SDG and ENL increased the expression of GPER both in vitro and in vivo. Furthermore, ENL-induced cell cycle arrest may be mediated by the activation of GPER/ERK pathway and subsequent upregulation of p53 and p21 and downregulation of cyclin D1. This work, in tandem with previous studies, will enhance our knowledge regarding the mechanism(s) of dietary phytochemicals on BPH prevention and ultimately expand the scope of adopting alternative approaches in BPH treatment.


cDNA cloning and characterization of a rat spermatogenesis-associated protein RSP29.
RSP29, a protein secreted by rat round spermatids, stimulates the secretory function of Sertoli cells in the testis. By making use of the N-terminal sequence homology of RSP29 and a human protein hDP1 that we had previously isolated, we cloned the full length cDNA sequence that encodes RSP29. The entire amino acid sequence of RSP29 showed significant homology with that of hDP1, which was later identified as glyoxalase II. Southern analysis showed that the RSP29 protein sequence is highly conserved in eukaryotes and possibly in prokaryotes. The RSP29 mRNA is expressed in many tissues but has an extremely high abundance in testis. These data suggest that RSP29 may have an important function in most tissues of enkaryotic organisms. The high expression of RSP29 in testis and its stimulatory effects on Sertoli cells suggest that RSP29 could be especially important in the regulation of spermatogenesis.


Deafening silence on a vital issue: The World Health Organization has ignored the sexuality of persons with disabilities.
In 2016, the World Health Organization, through the Global Cooperation on Assistive Technology Initiative, issued the Priority Assistive Products List which is meant to be a guide to member states of the 50 assistive products needed for a basic health care and/or social welfare system; it is also a model from which nations can develop their national priority assistive products lists. The aim of this opinion paper is to share my views about the Priority Assistive Products List on the grounds that it makes no distinct mention of sexual assistive devices, yet research has indicated that sexuality is an area of great concern for persons with disabilities. In any case, sexuality forms a core part of being human, and it impacts on both the physical and mental well-being of all human beings. I conclude in part that, in its present format, the list perpetuates the myth that persons with disabilities are asexual beings who are innocent of sexual thoughts, feelings and experiences. The list also propagates the stereotype that sexuality is a sacred, private, bedroom matter that should be kept out of the public domain, to the detriment of the health and well-being of persons with disabilities.


Fasting cycles retard growth of tumors and sensitize a range of cancer cell types to chemotherapy.
Short-term starvation (or fasting) protects normal cells, mice, and potentially humans from the harmful side effects of a variety of chemotherapy drugs. Here, we show that treatment with starvation conditions sensitized yeast cells (Saccharomyces cerevisiae) expressing the oncogene-like RAS2(val19) to oxidative stress and 15 of 17 mammalian cancer cell lines to chemotherapeutic agents. Cycles of starvation were as effective as chemotherapeutic agents in delaying progression of different tumors and increased the effectiveness of these drugs against melanoma, glioma, and breast cancer cells. In mouse models of neuroblastoma, fasting cycles plus chemotherapy drugs--but not either treatment alone--resulted in long-term cancer-free survival. In 4T1 breast cancer cells, short-term starvation resulted in increased phosphorylation of the stress-sensitizing Akt and S6 kinases, increased oxidative stress, caspase-3 cleavage, DNA damage, and apoptosis. These studies suggest that multiple cycles of fasting promote differential stress sensitization in a wide range of tumors and could potentially replace or augment the efficacy of certain chemotherapy drugs in the treatment of various cancers.


Evaluation of statistical inference on empirical resting state fMRI.
Modern statistical inference techniques may be able to improve the sensitivity and specificity of resting state functional magnetic resonance imaging (rs-fMRI) connectivity analysis through more realistic assumptions. In simulation, the advantages of such methods are readily demonstrable. However, quantitative empirical validation remains elusive in vivo as the true connectivity patterns are unknown and noise distributions are challenging to characterize, especially in ultra-high field (e.g., 7T fMRI). Though the physiological characteristics of the fMRI signal are difficult to replicate in controlled phantom studies, it is critical that the performance of statistical techniques be evaluated. The SIMulation EXtrapolation (SIMEX) method has enabled estimation of bias with asymptotically consistent estimators on empirical finite sample data by adding simulated noise . To avoid the requirement of accurate estimation of noise structure, the proposed quantitative evaluation approach leverages the theoretical core of SIMEX to study the properties of inference methods in the face of diminishing data (in contrast to increasing noise). The performance of ordinary and robust inference methods in simulation and empirical rs-fMRI are compared using the proposed quantitative evaluation approach. This study provides a simple, but powerful method for comparing a proxy for inference accuracy using empirical data.


Interspecies comparison of selected pollution biomarkers in dreissenid spp. inhabiting pristine and moderately polluted sites.
Stress biomarkers, which can outline impacts of contaminants in aquatic biota at the biochemical level, are increasingly used as early warning tools in environmental monitoring. Reliable biomarker based assessment schemes, however, request appropriate knowledge of baseline levels of selected endpoints, and the potential influence of a range of natural influencing factors (both abiotic and biotic) as well. In this study, we examined the interspecies variability of various biomarkers (metallothioneins (MT), ethoxyresorufin-O-deethylase activity (EROD), lipid peroxidation (LPO), DNA strand breaks (DNA_sb), vitellogenin-like proteins (Vtg)) in Dreissena polymorpha and Dreissena bugensis inhabiting either pristine- or moderately impacted sites of Lake Balaton (Hungary). Levels of all biomarkers considered revealed low interspecies variability in the two dreissenid species at all sampling sites, with consistently higher (but statistically insignificant) values in Dreissena polymorpha. Levels of all biomarkers varied within the two investigated seasons, with significant influence of the reproduction cycle particularly on the levels of metallothioneins and vitellogenin-like proteins. Each biomarker considered was elevated by October, with significantly higher values in the mussels inhabiting harbours. Insignificant spatial and temporal variability in the general health indicators (condition index, total protein content) of dreissenids was observed, which, in parallel with evident rise in biomarker levels, apparently suggest that the anthropogenic impacts in harbours affect mussel fitness yet at sub organismal level. Our data might serve useful basis for future environmental monitoring surveys, especially in habitats where the progressive replacement of Dreissena polymorpha by Dreissena bugensis is taking place, as the interspecies variability in susceptibility to chemical stress of the two species is well comparable.


Renin-Angiotensin system - considerations for hypertension and kidney.
The kidneys play a fundamental role in the long-term control of arterial pressure by regulating sodium balance and extracellular fluid volume. The renin-angiotensin system (RAS) is at the center of the regulation of hypertension and progressive renal injury. It has gradually become clear that not only systemic RAS, but also intrarenal RAS has specific effects in the pathogenesis and progression of hypertension and renal damage. All of the RAS components are exhibited in the kidney and intrarenal angiotensin II (Ang II) is formed by multiple mechanisms. The demonstration of much enhanced levels of Ang II within specific renal compartments points out selective local regulation of Ang II in the kidney, showing that intrarenal Ang II levels are regulated in a way different from circulating Ang II. The importance of the RAS in involving proper nephrogenesis is also well known, and suppression of the RAS during fetal development may play a key role in mediating the structural and physiological changes observed in models of fetal programming of hypertension.


Divergent evolution in M6P/IGF2R imprinting from the Jurassic to the Quaternary.
M6P/IGF2R imprinting first appeared approximately 150 million years ago following the divergence of prototherian from therian mammals. Although M6P/IGF2R is clearly imprinted in opossums and rodents, its imprint status in humans remains ambiguous. It is also still unknown if M6P/IGF2R imprinting was an ancestral mammalian epigenotype or if it evolved convergently. We report herein that M6P/IGF2R is imprinted in Artiodactyla, as it is in Rodentia and Marsupialia, but that it is not imprinted in Scandentia, Dermoptera and Primates, including ringtail lemurs and humans. These results are most parsimonious with a single ancestral origin of M6P/IGF2R imprinting followed by a lineage-specific disappearance of M6P/IGF2R imprinting in Euarchonta. The absence of M6P/IGF2R imprinting in extant primates, due to its disappearance from the primate lineage over 75 million years ago, demonstrates that imprinting at this locus does not predispose to human disease. Moreover, the divergent evolution of M6P/IGF2R imprinting predicts that the success of in vitro embryo procedures such as cloning may be species dependent.


Detection of Cryptosporidium parvum oocysts using a microfluidic device equipped with the SUS micromesh and FITC-labeled antibody.
Development of a microfluidic device equipped with micromesh for detection of Cryptosporidium parvum oocyst was reported. A micromesh consisting of 10 x 10 cavities was microfabricated on the stainless steel plate by laser ablation. Each cavity size, approximately 2.7 microm in diameter, was adopted to capture a single C. parvum oocyst. Under negative pressure operation, suspensions containing microbeads or C. parvum oocysts flowed into the microchannel. Due to strong non-specific adsorption of microbeads onto the PDMS microchannel surface during sample injection, the surface was treated with air plasma, followed by treatment with 1% sodium dodecyl sulfate (SDS) solution. This process reduced the non-specific adsorption of microbeads on the microchannel to 10% or less in comparison to a non-treated microchannel. This microfluidic device equipped with the SUS micromesh was further applied for the capture of C. parvum oocysts. Trapped C. parvum oocysts were visualized by staining with FITC-labeled anti-C. parvum oocyst antibody on a micromesh and counted under fluoroscopic observation. The result obtained by our method was consistent with that obtained by direct immunofluorescence assay coupled with immunomagnetic separation (DFA-IMS) method, indicating that the SUS micromesh is useful for counting of C. parvum oocysts. The newly designed microfluidic device exploits a geometry that allowed for the entrapment of oocysts on the micromesh while providing the rapid introduction of a series of reagents and washes through the microfluidic structure. Our data indicate that this microfluidic device is useful for high-throughput counting of C. parvum oocysts from tap water sample.


Aldosterone-secreting adrenocortical carcinomas are associated with unique operative risks and outcomes.
Adrenocortical carcinoma (ACC) that produces aldosterone is an extremely rare, uncharacterized endocrine malignancy. Our aim was to characterize this neoplasm in terms of its clinical behavior and patient outcomes. A retrospective review was made of all patients who had operative management of aldosterone-secreting ACC from 1957 to 2000 at the Mayo Clinic. Comparisons were made to patients with non-aldosterone-secreting ACC treated during the same period. Of 141 patients with ACC, we identified 15 patients with aldosterone-secreting ACC. Isolated aldosterone hypersecretion was present in 10 patients, and mixed hormonal secretion was detected in 5. Mean tumor size and weight were 10.8 cm and 453 g, respectively. Surgical management included curative resection in 10 patients (67%). Perioperative mortality was 20%. Disease recurred in 7 patients (70%) with a median interval of 17 months. Five-year survival was 52%. Patients with aldosterone-secreting ACC had an increased risk of perioperative mortality (20% vs 5%), yet they had an overall survival of 63 months compared to 19 months for patients with non-aldosterone-secreting ACC. Aldosterone hypersecretion occurs in 11% of all ACCs and is associated with unique operative risk and outcome. Although patients harboring aldosterone-secreting ACC appear to have an increased risk of perioperative death, survivors may have an improved overall survival rate compared with patients with non-aldosterone-secreting ACC.


A method for developing standardised interactive education for complex clinical guidelines.
Although systematic use of the Perinatal Society of Australia and New Zealand internationally endorsed Clinical Practice Guideline for Perinatal Mortality (PSANZ-CPG) improves health outcomes, implementation is inadequate. Its complexity is a feature known to be associated with non-compliance. Interactive education is effective as a guideline implementation strategy, but lacks an agreed definition. SCORPIO is an educational framework containing interactive and didactic teaching, but has not previously been used to implement guidelines. Our aim was to transform the PSANZ-CPG into an education workshop to develop quality standardised interactive education acceptable to participants for learning skills in collaborative interprofessional care. The workshop was developed using the construct of an educational framework (SCORPIO), the PSANZ-CPG, a transformation process and tutor training. After a pilot workshop with key target and stakeholder groups, modifications were made to this and subsequent workshops based on multisource written observations from interprofessional participants, tutors and an independent educator. This participatory action research process was used to monitor acceptability and educational standards. Standardised interactive education was defined as the attainment of content and teaching standards. Quantitative analysis of positive expressed as a percentage of total feedback was used to derive a total quality score. Eight workshops were held with 181 participants and 15 different tutors. Five versions resulted from the action research methodology. Thematic analysis of multisource observations identified eight recurring education themes or quality domains used for standardisation. The two content domains were curriculum and alignment with the guideline and the six teaching domains; overload, timing, didacticism, relevance, reproducibility and participant engagement. Engagement was the most challenging theme to resolve. Tutors identified all themes for revision whilst participants identified a number of teaching but no content themes. From version 1 to 5, a significant increasing trend in total quality score was obtained; participants: 55%, p=0.0001; educator: 42%, p=0.0004; tutor peers: 57%, p=0.0001. Complex clinical guidelines can be developed into a workshop acceptable to interprofessional participants. Eight quality domains provide a framework to standardise interactive teaching for complex clinical guidelines. Tutor peer review is important for content validity. This methodology may be useful for other guideline implementation.


Dynamic systems and the role of evaluation: The case of the Green Communities project.
The crucial role evaluation can play in the co-development of project design and its implementation will be addressed through the analysis of a case study, the Green Communities (GC) project, funded by the Italian Ministry of Environment within the EU Interregional Operational Program (2007-2013) "Renewable Energy and Energy Efficiency". The project's broader goals included an attempt to trigger a change in Italian local development strategies, especially for mountain and inland areas, which would be tailored to the real needs of communities, and based on a sustainable exploitation and management of the territorial assets. The goal was not achieved, and this paper addresses the issues of how GC could have been more effective in fostering a vision of change, and which design adaptations and evaluation procedures would have allowed the project to better cope with the unexpected consequences and resistances it encountered. The conclusions drawn are that projects should be conceived, designed and carried out as dynamic systems, inclusive of a dynamic and engaged evaluation enabling the generation of feedbacks loops, iteratively interpreting the narratives and dynamics unfolding within the project, and actively monitoring the potential of various relationships among project participants for generating positive social change.


Oral supplementations with free and dipeptide forms of L-glutamine in endotoxemic mice: effects on muscle glutamine-glutathione axis and heat shock proteins.
Sepsis is a leading cause of death in intensive care units worldwide. Low availability of glutamine contributes to the catabolic state of sepsis. L-Glutamine supplementation has antioxidant properties and modulates the expression of heat shock proteins (HSPs). This study investigated the effects of oral supplementation with L-glutamine plus L-alanine (GLN+ALA), both in the free form and L-alanyl-L-glutamine dipeptide (DIP), on glutamine-glutathione (GSH) axis and HSPs expression in endotoxemic mice. B6.129F2/J mice were subjected to endotoxemia (lipopolysaccharides from Escherichia coli, 5 mg.kg(-1), LPS group) and orally supplemented for 48 h with either L-glutamine (1 g.kg(-1)) plus L-alanine (0.61 g.kg(-1)) (GLN+ALA-LPS group) or 1.49 g.kg(-1) of DIP (DIP-LPS group). Endotoxemia reduced plasma and muscle glutamine concentrations [relative to CTRL group] which were restored in both GLN+ALA-LPS and DIP-LPS groups (P<.05). In supplemented groups were re-established GSH content and intracellular redox status (GSSG/GSH ratio) in circulating erythrocytes and muscle. Thiobarbituric acid reactive substance was 4-fold in LPS treated mice relative to the untreated CTRL group, and plasma TNF-α and IL-1β levels were attenuated by the supplements. Heat shock proteins 27, 70 and 90 (protein and mRNA) were elevated in the LPS group and were returned to basal levels (relative to CTRL group) in both GLN+ALA-LPS and DIP-LPS groups. Supplementations to endotoxemic mice resulted in up-regulation of GSH reductase, GSH peroxidase and glutamate cysteine ligase mRNA expression in muscle. In conclusion, oral supplementations with GLN+ALA or DIP are effective in reversing the conditions of LPS-induced deleterious impact on glutamine-GSH axis in mice under endotoxemia.


Glucose modulates Drosophila longevity and immunity independent of the microbiota.
The acquisition of nutrients is essential for maintenance of metabolic processes in all organisms. Nutritional imbalance contributes to myriad metabolic disorders that include malnutrition, diabetes and even cancer. Recently, the importance of macronutrient ratio of food has emerged as a critical factor to determine health outcomes. Here we show that individual modifications to a completely defined diet markedly impact multiple aspects of organism wellbeing in Drosophila melanogaster. Through a longitudinal survey of several diets we demonstrate that increased levels of dietary glucose significantly improve longevity and immunity in adult Drosophila. Our metagenomic studies show that relative macronutrient levels not only influence the host, but also have a profound impact on microbiota composition. However, we found that elevated dietary glucose extended the lifespan of adult flies even when raised in a germ-free environment. Furthermore, when challenged with a chronic enteric infection, flies fed a diet with added glucose had increased survival times even in the absence of an intact microbiota. Thus, in contrast to known links between the microbiota and animal health, our findings uncover a novel microbiota-independent response to diet that impacts host wellbeing. As dietary responses are highly conserved in animals, we believe our results offer a general understanding of the association between glucose metabolism and animal health.


Life cycle of a minimal protocell--a dissipative particle dynamics study.
Cross-reactions and other systematic difficulties generated by the coupling of functional chemical subsystems pose the largest challenge for assembling a viable protocell in the laboratory. Our current work seeks to identify and clarify such key issues as we represent and analyze in simulation a full implementation of a minimal protocell. Using a 3D dissipative particle dynamics simulation method, we are able to address the coupled diffusion, self-assembly, and chemical reaction processes required to model a full life cycle of a protocell composed of coupled genetic, metabolic, and container subsystems. Utilizing this minimal structural and functional representation of the constituent molecules, their interactions, and their reactions, we identify and explore the nature of the many linked processes for the full protocellular system. Obviously the simplicity of this simulation method combined with the inherent system complexity prevents us from expecting quantitative simulation predictions from these investigations. However, we report important findings on systemic processes, some previously predicted and some newly discovered, as we couple the protocellular self-assembly processes and chemical reactions.


Contours in noise: a role for self-cuing?
A contour formed of aligned Gabor patches is easily detected amidst a dense background of randomly oriented patches; the longer the string of aligned patches, the more easily the contour is detected (D. J. Field, A. Hayes, & R. F. Hess, 1993). Here we show that a short string of collinear elements acts as a cue pointing to other patches of similar orientation that lie along the path defined by the string. Cueing might increase the gain of similarly oriented elements in the vicinity and/or reduce the number of potential locations (uncertainty) that the observer monitors. To assess the strength of the contour cue, I measured sensitivity to contrast increments on a test patch placed at various offsets relative to the cueing contour. Noise density and the length of cueing contour were also manipulated. Signal detection theory analysis of the psychometric function provided estimates of the gain and uncertainty parameters associated with each condition. In the presence of noise, observers were best at detecting a contrast increment on a patch that was aligned with the cueing string. Gain estimates were high for the aligned condition but decreased with increasing offset from this position. Uncertainty estimates were invariant with offset at a given noise level but increased with increasing noise. Uncertainty also decreased significantly when the length of the cueing string was increased. The high gain for collinear test patches and the ability of the contour to reduce uncertainty at high noise densities is indeed similar to the effect of an explicit cue (e.g., Z.-L. Lu & B. A. Dosher, 1998), suggesting that one part of the contour cues other parts and contributes to the detectability of contours in noise.


Country-level and individual correlates of overweight and obesity among primary school children: a cross-sectional study in seven European countries.
The present study aims to estimate childhood overweight and obesity prevalence and their association with individual and population-level correlates in Eastern and Western European countries. Data were obtained from the School Children Mental Health in Europe, a cross-sectional survey conducted in 2010 in Italy, Germany, the Netherlands, Romania, Bulgaria, Lithuania and Turkey. The sample consists of 5,206 school children aged 6 to 11 years old. Information on socio-demographics, children's height and weight, life-style and parental attitude were reported by the mothers. Country-level indicators were obtained through several data banks. Overweight and obesity in children were calculated according to the international age and gender-specific child Body Mass Index cut-off points. Multivariable logistic regression models included socio-demographic, lifestyle, mothers' attitude, and country-level indicators to examine the correlates of overweight. Overall prevalence was 15.6% (95% CI = 19.3-21.7%) for overweight and 4.9% (95% CI = 4.3-5.6%) for obesity. In overweight (including obesity), Romanian children had the highest prevalence (31.4%, 95% CI = 28.1-34.6%) and Italian the lowest (10.4%, 95% CI = 8.1-12.6%). Models in the pooled sample showed that being younger (aOR = 0.93, 95% = CI 0.87-0.97), male (aOR = 1.24, 95% CI = 1.07-1.43), an only child (aOR = 1.40, 95% CI = 1.07-1.84), spending more hours per week watching TV (aOR = 1.01, 95% CI =1.002-1.03), and living in an Eastern Country were associated with greater risk of childhood overweight (including obesity). The same predictors were significantly associated with childhood overweight in the model conducted in the Eastern region, but not in the West. Higher Gross Domestic Product and Real Domestic Product, greater number of motor and passenger vehicles, higher percentage of energy available from fat, and more public sector expenditure on health were also associated with lower risk for childhood overweight after adjusting for covariables in the pooled sample and in the east of Europe, but not in the West. Prevalence rates of overweight and obesity in school children is still high, especially in Eastern regions, with some socio-demographic factors and life-styles associated with being overweight. It is also in the Eastern region itself where better macro-economic indicators are related with lower rates of childhood overweight. This represents a public health concern that deserves special attention in those countries undertaking economic and political transitions.


The use of artificial intelligence and robotics in regional anaesthesia.
The current fourth industrial revolution is a distinct technological era characterised by the blurring of physics, computing and biology. The driver of change is data, powered by artificial intelligence. The UK National Health Service Topol Report embraced this digital revolution and emphasised the importance of artificial intelligence to the health service. Application of artificial intelligence within regional anaesthesia, however, remains limited. An example of the use of a convoluted neural network applied to visual detection of nerves on ultrasound images is described. New technologies that may impact on regional anaesthesia include robotics and artificial sensing. Robotics in anaesthesia falls into three categories. The first, used commonly, is pharmaceutical, typified by target-controlled anaesthesia using electroencephalography within a feedback loop. Other types include mechanical robots that provide precision and dexterity better than humans, and cognitive robots that act as decision support systems. It is likely that the latter technology will expand considerably over the next decades and provide an autopilot for anaesthesia. Technical robotics will focus on the development of accurate sensors for training that incorporate visual and motion metrics. These will be incorporated into augmented reality and visual reality environments that will provide training at home or the office on life-like simulators. Real-time feedback will be offered that stimulates and rewards performance. In discussing the scope, applications, limitations and barriers to adoption of these technologies, we aimed to stimulate discussion towards a framework for the optimal application of current and emerging technologies in regional anaesthesia.


An epidemiological and clinical study of transaminase levels and hepatitis B antibodies in 1,100 blood donors.
In order to provide epidemiological and clinical information on surrogate testing of blood donations, the respective prevalences of serum hepatitis B virus (HBV) markers and elevated transaminase levels were studied in 1,100 blood donors according to their geographic origin and socioeconomic level. The frequency of serum HBV markers varied as a function of HBV endemicity in the country of origin; however, it was inversely correlated (p less than 0.05) to the socioeconomic level of the donors, even in those originating from countries of low HBV endemicity. There was no association between serum HBV markers and the increased transaminase level which was observed in 48 (4.3%) donors. Twenty-five of these accepted further clinical evaluation. A diagnosis appeared probable in 12 of the 25: alcohol in 5; drugs in 6; non-A, non-B hepatitis in 1. Seven of the remaining 13 subjects were more than 25% above ideal body weight. Transaminase activities determined at the time of clinical assessment were normal in 14. In addition, serum HBV DNA was found in 5 of 247 donors, even in the absence of any usual HBV marker and/or hypertransaminasemia. This could account for the few cases of B and B-like posttransfusion hepatitis which are known to still occur despite careful HBsAg screening of blood donors.


Mitigating seabird bycatch during hauling by pelagic longline vessels.
Bycatch in longline fisheries threatens the viability of some seabird populations. The Hawaii longline swordfish fishery reduced seabird captures by an order of magnitude primarily through mitigating bycatch during setting. Now, 75% of captures occur during hauling. We fit observer data to a generalized additive regression model with mixed effects to determine the significance of the effect of various factors on the standardized seabird haul catch rate. Density of albatrosses attending vessels during hauling, leader length and year had largest model effects. The standardized haul catch rate significantly increased with increased albatross density during hauling. The standardized catch rate was significantly higher the longer the leader: shorter leaders place weighted swivels closer to hooks, reducing the likelihood of baited hooks becoming available to surface-scavenging albatrosses. There was a significant linear increasing temporal trend in the standardized catch rate, possibly partly due to an observed increasing temporal trend in the local abundance of albatrosses attending vessels during hauling. Swivel weight, Beaufort scale and season were also significant but smaller model effects. Most (81%) haul captures were on branchlines actively being retrieved. Future haul mitigation research should therefore focus on reducing bird access to hooks as crew coil branchlines, including methods identified here of shorter leaders and heavier swivels, and other potentially effective methods, including faster branchline coiling and shielding the area where hooks becomes accessible. The proportion of Laysan albatross (Phoebastria immutabilis) captures that occurred during hauling was significantly, 1.6 times, higher than for black-footed albatrosses (P. nigripes), perhaps due to differences in the time of day of foraging and in daytime scavenging competitiveness; mitigating haul bycatch would therefore be a larger benefit to Laysans. Locally, findings identify opportunities to nearly eliminate seabird bycatch. Globally, findings fill a gap in knowledge of methods to mitigate seabird bycatch during pelagic longline hauling.


Key metrics and processes for validating embryo diagnostics.
Embryo diagnostics are somewhat controversial in clinical assisted reproduction technology (ART) practice and remain an active area of investigation. Application of embryo diagnostics holds great potential to raise the standard of clinical care by eliminating futile transfers, allowing highly effective single-embryo transfer, and reducing the probability of clinical loss and ongoing abnormal gestations. These advantages are accompanied by risks, principally the chance that a reproductively competent embryo will be mislabeled and discarded. This would lower the ultimate probability that one or more of the embryos might implant and lead to delivery of a healthy infant. Rigorous validation should be required for embryo diagnostics. Metrics for validation can be divided into three simple areas: analytical validation, determination of clinical predictive values for normal and abnormal test results, and a randomized clinical trial to demonstrate that the selection advantage gained through the diagnostic improves clinical outcomes.


Ocular ultrasound to detect intracranial hypertension in trauma patients.
Increases in intracranial pressure (ICP) after head trauma require a rapid recognition to allow for adequate treatments. The aim of this study was to determine whether dilation of the optic nerve sheath, as detected by ocular ultrasound at the bedside, could reliably identify increases in ICP assessed with an intraparenchymal probe in adult head trauma patients. Eleven head trauma injured adult patients admitted to the intensive care unit with a Glasgow Coma Scale score ≤8, with cerebral contusion confirmed by computed tomography scan, and that required invasive ICP monitoring, were enrolled in the study. ICP values ≤20 mm Hg were considered as normal. Patients with acute or chronic ocular lesion were excluded. Ten nontrauma intensive care unit patients, with no ICP monitoring, were enrolled as control group. Invasive arterial pressure was monitored, and optic nerve sheath diameter (ONSD) was assessed by ocular ultrasound in all the patients. Head trauma patients without intracranial hypertension had ONSD values, assessed by ultrasound, equivalent to those measured in control patients (5.52 mm ± 0.36 mm vs. 5.51 mm ± 0.32 mm). ONSD, instead, significantly increased to 7.0 mm ± 0.58 mm, when ICP rose in value to >20 mm Hg (p < 0.0001 vs. normal ICP and control). ONSD values were significantly correlated to ICP values (r = 0.74, p < 0.001). When ICP was higher than 20 mm Hg, the ONSD diameter increased, whereas when the ICP was below 20 mm Hg, the ONSD returned to values equivalent to those assessed in control nontrauma patients. Accordingly, ocular ultrasound may be considered as a good alternative for a rapid indirect evaluation of head trauma patients' ICP.


Establishing Caenorhabditis elegans as a model for Mycobacterium avium subspecies hominissuis infection and intestinal colonization.
The nematode Caenorhabditis elegans has become a model system for studying the disease interaction between pathogens and the host. To determine whether the transparent nematode could serve as a useful model for Mycobacterium avium subspecies hominissuis (MAH) infection of the intestinal tract, worms were fed MAH and assayed for the effects of the bacterial infection on the worm. It was observed during feeding that viable MAH increases in the intestinal lumen in a time dependent manner. Ingestion of MAH was deemed non-toxic to worms as MAH-fed populations have similar survival curves to those fed E. coli strain OP50. Pulse-chase analysis using E. coli strain OP50 revealed that MAH colonize the intestinal tract, as viable MAH remain within the intestine after the assay. Visualization of intestinal MAH using histology and transmission electron microscopy demonstrates that MAH localizes to the intestinal lumen, as well as establishes direct contact with intestinal epithelium. Bacterial colonization appears to have a detrimental effect on the microvilli of the intestinal epithelial cells. The MAH ΔGPL/4B2 strain with a mutation in glycopeptidolipid production is deficient in binding to human epithelial cells (HEp-2), as well as deficient in its ability to bind to and colonize the intestinal tract of C. elegans as efficiently as wild-type MAH. These data indicate the C. elegans may serve as a useful model system for MAH pathogenesis and in determining the mechanisms used by MAH during infection and colonization of the intestinal epithelium.


Church attendance and new episodes of major depression in a community study of older adults: the Cache County Study.
We examined the relation between church attendance, membership in the Church of Jesus Christ of Latter-Day Saints (LDS), and major depressive episode, in a population-based study of aging and dementia in Cache County, Utah. Participants included 2,989 nondemented individuals aged between 65 and 100 years who were interviewed initially in 1995 to 1996 and again in 1998 to 1999. LDS church members reported twice the rate of major depression that non-LDS members did (odds ratio = 2.56, 95% confidence interval = 1.07-6.08). Individuals attending church weekly or more often had a significantly lower risk for major depression. After controlling for demographic and health variables and the strongest predictor of future episodes of depression, a prior depression history, we found that church attendance more often than weekly remained a significant protectant (odds ratio = 0.51, 95% confidence interval = 0.28-0.92). Results suggest that there may be a threshold of church attendance that is necessary for a person to garner long-term protection from depression. We discuss sociological factors relevant to LDS culture.


Center of plantar pressure during gait in pregnancy-related pelvic girdle pain and the effect of pelvic belts.
Many pregnant women suffer from pelvic girdle pain (PGP) during pregnancy. Etiologies are multifactorial and affect the joint stability of the sacroiliac joint. Pelvic belts could restore stability and reduce pain during gait. The center of pressure (COP) is a reliable parameter to assess gait and balance. The objectives of this study were to analyze the COP during gait in pregnant women with PGP, to evaluate the effect of pelvic belts and to compare two types of belts on COP parameters. 46 pregnant women with PGP, 58 healthy pregnant women and 23 non-pregnant women participated in the study. The motor task consisted of three gait trials at different velocities on an electronic walkway. Two pelvic belts for pregnant women were used. An analysis of variance was performed to determine the effects of the progression of the pregnancy, gait speed, presence of pregnancy and occurrence of pain on the COP parameters. Compared to the control group, pregnant women with PGP had a higher stance time, but COP displacement and velocity were lower. The COP parameters varied between pregnant women with and without pelvic girdle pain, the use of a belt during pregnancy decreased the walking velocity. No difference was found according to the type of belt. Differences in COP parameters during gait between pregnant women with or without PGP were minimal. Pelvic girdle pain did not affect the center of pres- sure. Wearing a belt during pregnancy modified the center of pressure velocity during gait in pregnant women with PGP.


Exploration of the association between apology and forgiveness amongst victims of human rights violations.
Forgiving may lead to an improvement of mental health, and from a therapeutic jurisprudence perspective it is important to establish what aspects of judicial procedures can be changed to promote forgiving. The literature suggests that receiving an apology may encourage forgiving. However, there is a dearth of empirical research regarding the association between forgiving and apology in judicial settings. This paper reports the findings of a study that examined the association between forgiving and four restorative situations (i.e. excuse, admission of guilt, apology, and true sorriness) in a group of 134 victims of gross human rights violations who were actual or potential participants in the proceedings of the South African Truth and Reconciliation Commission. The best predictors of forgiveness in this sample were gender and whether victims perceived wrongdoers to be truly sorry.


Nanoparticle synthesis via bubbling vapor precursors in bulk liquids.
Conventional methods for preparing polymer nanoparticles and organic-inorganic composite nanoparticles use solution based top-down processes with surfactants and mechanical stirring. Examples of such processes include emulsion polymerization of monomers to produce polymer nanoparticles and sol-gel reactions involving hydrolysis of inorganic precursors to produce inorganic materials (such as silica and titanium nanoparticles). Here, we show that vaporized precursors of various compounds can be used as reactants to produce polymer, inorganic, and composite nanoparticles. The bubbling action of precursor vapor in a reactant vessel provides a constant supply of precursor species while aiding their rapid mixing in the bulk solution liquid. The vaporization and bubbling processes require only small amounts of energy to prepare nanoparticles or core-shell nanoparticles without forming unwanted side products. Compared to other available techniques, this approach enables precise control of nanoparticle size and shell thickness as functions of vapor supply time and temperature without surfactants. Our approach can potentially be applied to fabricate functional nanomaterials using organic and inorganic precursors for medical, electrical, optical, magnetic and/or catalytic applications.


Serial lumbar CSF presure measurements and cranial computed tomographic findings in childhood tuberculous meningitits.
Intracranial pressure (ICP) was monitored in 218 consecutive children with hydrocephalus secondary to tuberculous meningitis (TBM). All children underwent cranial computerized tomographic (CT) scanning and continuous lumbar cerebrospinal (CSF) pressure monitoring on admission. Noncommunicating hydrocephalus (37 children), as determined by air encephalography, was treated by ventriculoperitoneal (VP) shunting and communicating hydrocephalus (181 children), by means of daily acetazolamide and frusemide. Response of ICP to treatment in the group with communicating hydrocephalus was assessed by means of repeated CSF pressure monitoring and CT scanning. One hundred and eighty-five of the 218 patients survived the 1st month of treatment. The aim of this study was the retrospective determination of (1) the relationship between ICP measurements and CT findings on admission and (2) the characteristics of the ICP recording which correlated best with the CT criteria of compensated hydrocephalus after the 1st month of treatment. No relationship was found between the level of baseline CSF pressure and the degree of hydrocephalus, as demonstrated by CT scanning, on admission. Seventy-five per cent of the patients with communicating hydrocephalus that survived the 1st month of treatment complied with the CT criteria for compensated hydrocephalus. All these patients had a baseline CSF pressure below 15 mmHg and absence of high-amplitude B waves on the pressure recording done at the end of the 1st month. In this study repeated lumbar CSF pressure monitoring proved to be an effective instrument to assess the response of communicating tuberculous hydrocephalus to medical treatment and also accurately predicted the timing of compensation of the hydrocephalus.


Mammalian spinal biomechanics: postural support in seated macaques.
The aim of this study was to investigate whether the ligamentous lumbar vertebral column of a macaque could potentially provide passive mechanical support to the weight of the head, upper body and forelimbs during upright sitting. The seated flexed curvature of the lumbar spine of Macaca sylvana was estimated from a photograph and was partitioned equally among the lumbar-lumbar intervertebral joints. This flexed curvature was compared with the hyper-extended profile of the unloaded excised ligamentous spine of a related species (Macaca fascicularis) and used to calculate changes in intervertebral angle from the unloaded excised state to the loaded in vivo state. Changes in intervertebral angle were then used to calculate the net flexion moment required to bend the spine from the unloaded curvature to the seated curvature. The moment arm of the ventrally displaced weight of the head, upper body and forelimbs was estimated and used to calculate a corresponding net force. It was found that this force corresponded to approximately 18 % of the total body weight of the 2.34 kg sample animal. This compares with a likely fractional body weight of approximately 30-40 % for the head, upper body and forelimbs of these primates. Therefore, approximately half of the ventral flexion moment associated with the combined weight of the head, upper body and forelimbs during sitting in these animals may be supported by the passive mechanical properties associated with the ligamentous lumbar spine. This represents a potential means of relieving muscular effort and saving metabolic energy.


Delocalized and localized donating-accepting Mn-C interactions in half-sandwich cyclopentadienyl and pentadienyl complexes.
The widely used cyclopentadienyl (Cp) ligand and the pentadienyl (Pdl) ligand have very similar molecular orbitals (MOs) but they have significantly different chemical behavior. The extensively mixing of the MOs in the Pdl ligand has prevented a quantitative explanation of the observed differences between these two ligands. In a series of synthesized close/open half sandwich manganese phosphine carbonyls, the quantum theory of atoms in molecules and the interacting quantum atom model allow us to quantify the accepting-donating capacities of both ligands and to break down the M-dienyl bond into individual atomic interactions. The Mn-Pdl bond is stronger, more localized and has a larger backbonding contribution than the Mn-Cp bond. All carbon atoms in the Cp ring interact similarly with the Mn center, whereas the Pdl's terminal carbon atoms exhibit a strong donor and a moderate acceptor character, and C2 and C4 have an acceptor character and C3 a donor character. Contrarily to the constrained five-membered Cp ring, the stronger Mn-Pdl interaction can barely compensate for the large destabilizing distortion of the backbone of the nonaromatic Pdl ligand.


A functional MRI marker may predict the outcome of electroconvulsive therapy in severe and treatment-resistant depression.
Electroconvulsive therapy (ECT) is effective even in treatment-resistant patients with major depression. Currently, there are no markers available that can assist in identifying those patients most likely to benefit from ECT. In the present study, we investigated whether resting-state network connectivity can predict treatment outcome for individual patients. We included forty-five patients with severe and treatment-resistant unipolar depression and collected functional magnetic resonance imaging scans before the course of ECT. We extracted resting-state networks and used multivariate pattern analysis to discover networks that predicted recovery from depression. Cross-validation revealed two resting-state networks with significant classification accuracy after correction for multiple comparisons. A network centered in the dorsomedial prefrontal cortex (including the dorsolateral prefrontal cortex, orbitofrontal cortex and posterior cingulate cortex) showed a sensitivity of 84% and specificity of 85%. Another network centered in the anterior cingulate cortex (including the dorsolateral prefrontal cortex, sensorimotor cortex, parahippocampal gyrus and midbrain) showed a sensitivity of 80% and a specificity of 75%. These preliminary results demonstrate that resting-state networks may predict treatment outcome for individual patients and suggest that resting-state networks have the potential to serve as prognostic neuroimaging biomarkers to guide personalized treatment decisions.


Specialized morphology corresponds to a generalist diet: linking form and function in smashing mantis shrimp crustaceans.
Many animals are considered to be specialists because they have feeding structures that are fine-tuned for consuming specific prey. For example, "smasher" mantis shrimp have highly specialized predatory appendages that generate forceful strikes to break apart hard-shelled prey. Anecdotal observations suggest, however, that the diet of smashers may include soft-bodied prey as well. Our goal was to examine the diet breadth of the smasher mantis shrimp, Neogonodactylus bredini, to determine whether it has a narrow diet of hard-shelled prey. We combined studies of prey abundance, feeding behavior, and stable isotope analyses of diet in both seagrass and coral rubble to determine if N. bredini's diet was consistent across different habitat types. The abundances of hard-shelled and soft-bodied prey varied between habitats. In feeding experiments, N. bredini consumed both prey types. N. bredini consumed a range of different prey in the field as well and, unexpectedly, the stable isotope analysis demonstrated that soft-bodied prey comprised a large proportion (29-53 %) of the diet in both habitats. Using a Bayesian mixing model framework (MixSIAR), we found that this result held even when we used uninformative, or generalist, priors and informative priors reflecting a specialist diet on hard-shelled prey and prey abundances in the field. Thus, contrary to expectation, the specialized feeding morphology of N. bredini corresponds to a broad diet of both hard-shelled and soft-bodied prey. Using multiple lines of study to describe the natural diets of other presumed specialists may demonstrate that specialized morphology often broadens rather than narrows diet breadth.


Up-regulation of MUC5AC mRNA expression in endotoxin-induced otitis media.
We investigated the expression levels of MUC5AC in endotoxin-induced otitis media with effusion (OME) in the rat using competitive polymerase chain reaction (PCR) and the morphology of middle ear mucosa using transmission electron microscopy (TEM). Experimental OME in the rat was induced after middle ear instillation of Escherichia coli lipopolysaccharides (LPS). Middle ear mucosa were obtained at 0 h, 12 h, Day 1, Day 3, Day 7 and Day 14 and reverse transcriptase (RT)-PCRs were then performed for the identification of MUC1, MUC2, MUC5AC and submandibular mucin 1 expression, followed by competitive PCRs for MUC5AC and beta2-microglobulin expression. Normal middle ear mucosa revealed no expression of mucin genes, whereas endotoxin upregulated the expression of MUC5AC mRNA between 12 h and Day 7, with maximal expression at Days 1 and 3. Middle ears treated three times with LPS upregulated more MUC5AC mRNA expression, by a factor of approximately 3.5, than those 1 day after one instillation. On TEM, dark granulated cells were observed at Day 3 after endotoxin instillation, but mixed granulated cells were seen on the ears treated three times with LPS. These results suggest that MUC5AC could be one of the major mucin genes in the middle ear mucosa related to otitis media.


Numerical emulation of Thru-Reflection-Line calibration for the de-embedding of Surface Acoustic Wave devices.
In this contribution, a rigorous numerical calibration is proposed to characterize the excitation of propagating mechanical waves by interdigitated transducers (IDTs). The transition from IDT terminals to phonon waveguides is modeled by means of a general circuit representation that makes use of Scattering Matrix (SM) formalism. In particular, the three-step calibration approach called the Thru-Reflection-Line (TRL), that is a well-established technique in microwave engineering, has been successfully applied to emulate typical experimental conditions. The proposed procedure is suitable for the synthesis/optimization of surface-acoustic-wave (SAW) based devices: the TRL calibration allows to extract/de-embed the acoustic component, namely resonator or filter, from the outer IDT structure, regardless of complexity and size of the letter. We report, as a result, the hybrid scattering parameters of the IDT transition to a mechanical waveguide formed by a phononic crystal patterned on a piezoelectric AlN membrane, where the effect of a discontinuity from periodic to uniform mechanical waveguide is also characterized. In addition, to ensure the correctness of our numerical calculations, the proposed method has been validated by independent calculations.


Impact of health education on community knowledge, attitudes and behaviour towards solid waste management in Al Ghobeiry, Beirut.
The risks posed by accumulation of solid waste are most obvious in developing countries, where waste collection and treatment is often inadequate. This study aimed to determine the impact of a health education intervention (based on lectures and focus group discussions) on community knowledge, attitudes and behaviours concerning solid waste management in Al Ghobeiry, Beirut. A randomly selected sample of 320 inhabitants were divide into intervention and control groups who completed the same questionnaire in the pre- and post-intervention phases. Compared with the control group the intervention group, who attended the health education sessions, showed: significantly better knowledge about the problems of and diseases spread by accumulation of solid waste; better attitudes to management of solid waste collection; and improved practices in terms of handling and recycling of household waste. There was an observed increased participation by people in cleaning campaigns and voluntary work in all the municipality activities.


Disappearance of female genital mutilation from the Bedouin population of Southern Israel.
Recently, clinicians in Southern Israel perceived that the practice of female genital mutilation had disappeared entirely in the Bedouin population. We previously studied the prevalence of this practice in 1995. We decided to survey again the Bedouin population focusing on those tribes previously reported to perform this practice. Eighty percent of the interviews were done by an Arabic-speaking psychiatrist and 20% were done by an Arabic speaking nurse in the gynecologic clinic of a large Bedouin township or the gynecologic clinic of a smaller Bedouin township. Women were asked if they would be willing to answer a few questions about their past and if they were willing to have the gynecologist, with no additional procedure, note whether any operation had been performed on their genitalia. Physical examination by gynecologist and an oral questionnaire. One hundred and thirty two women were examined. No cases of any scarring of the kind reported in the previous study were found on physical examination. FGM has apparently disappeared over 15 years in a population in which it was once prevalent.


[Application of " Internet Plus" AIDS prevention services among men who have sex with men in Guangzhou, China: results from 2010 to 2015].
Objective: To introduce the development strategy of " Internet Plus" AIDS prevention services and its implementation results from 2010 to 2015 in Guangzhou, China. Methods: A gay men's health column was created for an active website aimed at men who have sex with men(MSM), in collaboration with local community organizations and the Guangzhou CDC. We designed intervention tools(including scenario-based applications and HIV risk self-assessment systems)and an online HIV testing service platform, integrated with applied psychology and behavioral theory as well as the " Internet Plus" concept, to intervene in HIV infection risk factors among MSM. Data of clients who accessed the " Internet Plus" AIDS services from 2010 to 2015 were used to evaluate service operation. Six-year consecutive surveys, conducted between April and July of each service year, were collected using a national AIDS sentinel surveillance questionnaire. For each year of surveillance, information on HIV prevalence, HIV interventions received during the past year, unprotected anal intercourse in the past 6 months, and HIV testing in the past year were compared using the chi-squared(χ2)test, to roughly reflect the effect of"Internet Plus" AIDS prevention services. Results: As of 31 December 2015, a total of 34 395 MSM had received " Internet Plus" services and HIV testing. The number of MSM tested increased from 2 338 in 2010 to 8 054 in 2015. From 2010 to 2015, newly identified HIV cases in each year were 59, 166, 312, 283, 291, and 270, which accounted for 25.0%, 32.8%, 38.8%, 35.1%, 30.5%, and 23.2% of MSM HIV cases of Guangzhou, respectively. Sentinel surveillance data showed that during the study period, 3 047 MSM were investigated, with 405, 400, 401, 633, 608, and 600 each year, respectively. The proportion of participants who had received any HIV intervention during the past year was 74.3%(301), 70.8%(283), 83.3%(334), 85.0%(538), 69.1%(420), and 83.8%(503)each year, respectively(trend χ2=6.53, P=0.011). HIV testing done during the past year accounted for 44.0%(178), 44.3%(177), 49.4%(198), 53.4%(338), 56.1%(341), and 60.2%(361)each year, respectively(trend χ2=40.83, P<0.001). Unprotected anal intercourse in the past 6 months accounted for 59.3%(240), 62.0%(248), 56.6%(227), 57.0%(361), 48.4%(294), and 43.7%(262)each year, respectively(trend χ2=42.21, P<0.001). Conclusion: The"Internet Plus"AIDS prevention services in this study represent a manner to enhance traditional HIV prevention strategies. We found these services to be effective in implementation of the national AIDS control and prevention strategy, especially for the expansion of intervention, testing, and case identification among high-risk populations.


Uncoupling of T-cell effector functions by inhibitory killer immunoglobulin-like receptors.
Killer immunoglobulin-like receptors (KIRs) are a family of regulatory cell-surface molecules expressed on natural killer (NK) cells and memory T-cell subsets. Their ability to prevent the formation of an activation platform and to inhibit NK cell activation is the basis of the missing self model of NK cell function. The benefits of KIR expression for T-cell biology are unclear. We studied how KIR2DL2 regulates T-cell function. Engagement of KIR2DL2 by the ligand human leukocyte antigen (HLA)-Cw3 did not affect conjugate formation between CD4(+)KIR2DL2(+) T cells and superantigen-pulsed target cells or the development of mature immune synapses with lipid rafts. KIR2DL2 and the corresponding HLA-C ligand were initially recruited to the peripheral supramolecular activation cluster (pSMAC). Consequently, KIR2DL2 engagement did not inhibit the phosphorylation of early signaling proteins and T-cell-receptor (TCR)-mediated cytotoxicity or granule exocytosis. After 15-30 minutes, KIR2DL2 moved to the central supramolecular activation cluster (cSMAC), colocalizing with CD3. TCR synapses dissociated, and phosphorylated phospholipase C (PLC)-gamma1, Vav1, and extracellular signal-regulated kinase 1/2 (ERK1/2) were reduced 90 minutes after stimulation. Gene array studies documented that the inhibition of late signaling events by KIR2DL2 affected transcriptional gene activation. We propose that KIRs on memory T cells operate to uncouple effector functions by modifying the transcriptional profile while leaving granule exocytosis unabated.


Between Descriptors and Properties: Understanding the Ligand Efficiency Trends for G Protein-Coupled Receptor and Kinase Structure-Activity Data Sets.
The chemical meaning of the ligand efficiency (LE) metrics is explained in this paper using a large G protein-coupled receptor (GPCR) and kinase structure-activity (IC50, Ki) data set. Although there is a controversy in the literature regarding both the mathematical validity and the performance of LE, it is in common use as an early estimator for drug optimization. Apparently, the numerous con arguments are not convincing enough. We show here for the first time that the main misunderstanding of the chemical meaning of LE is its interpretation as a molecular descriptor connected with a single molecule. Instead, LE should be interpreted as a statistical property. We show that the LE, which is designed as a regression of a binding property on the heavy atom count (HAC), is correlated to the reciprocal of the molecular weight because of Avogadro statistics. This indicates that the hyperbolic model of LE is basically a consequence of a nonbinding effect, an increase in the number of ligands that are available to a receptor for smaller molecules, and not a real increase in the binding potency for a single HAC as interpreted in the literature. Accordingly, we need to revisit and carefully reevaluate LE-based molecular comparisons.


Subthalamic Nucleus Stimulation-Induced Local Field Potential Changes in Dystonia.
Subthalamic nucleus (STN) stimulation is an effective treatment for Parkinson's disease and induced local field potential (LFP) changes that have been linked with clinical improvement. STN stimulation has also been used in dystonia although the internal globus pallidus is the standard target where theta power has been suggested as a physiomarker for adaptive stimulation. We aimed to explore if enhanced theta power was also present in STN and if stimulation-induced spectral changes that were previously reported for Parkinson's disease would occur in dystonia. We recorded LFPs from 7 patients (12 hemispheres) with isolated craniocervical dystonia whose electrodes were placed such that inferior, middle, and superior contacts covered STN, zona incerta, and thalamus. We did not observe prominent theta power in STN at rest. STN stimulation induced similar spectral changes in dystonia as in Parkinson's disease, such as broadband power suppression, evoked resonant neural activity (ERNA), finely-tuned gamma oscillations, and an increase in aperiodic exponents in STN-LFPs. Both power suppression and ERNA localize to STN. Based on this, single-pulse STN stimulation elicits evoked neural activities with largest amplitudes in STN, which are relayed to the zona incerta and thalamus with changing characteristics as the distance from STN increases. Our results show that STN stimulation-induced spectral changes are a nondisease-specific response to high-frequency stimulation, which can serve as placement markers for STN. This broadens the scope of STN stimulation and makes it an option for other disorders with excessive oscillatory peaks in STN. © 2022 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.


Peritoneal GATA6+ macrophages function as a portal for Staphylococcus aureus dissemination.
Essentially all Staphylococcus aureus (S. aureus) bacteria that gain access to the circulation are plucked out of the bloodstream by the intravascular macrophages of the liver - the Kupffer cells. It is also thought that these bacteria are disseminated via the bloodstream to other organs. Our data show that S. aureus inside Kupffer cells grew and escaped across the mesothelium into the peritoneal cavity and immediately infected GATA-binding factor 6-positive (GATA6+) peritoneal cavity macrophages. These macrophages provided a haven for S. aureus, thereby delaying the neutrophilic response in the peritoneum by 48 hours and allowing dissemination to various peritoneal and retroperitoneal organs including the kidneys. In mice deficient in GATA6+ peritoneal macrophages, neutrophils infiltrated more robustly and reduced S. aureus dissemination. Antibiotics administered i.v. did not prevent dissemination into the peritoneum or to the kidneys, whereas peritoneal administration of vancomycin (particularly liposomal vancomycin with optimized intracellular penetrance capacity) reduced kidney infection and mortality, even when administered 24 hours after infection. These data indicate that GATA6+ macrophages within the peritoneal cavity are a conduit of dissemination for i.v. S. aureus, and changing the route of antibiotic delivery could provide a more effective treatment for patients with peritonitis-associated bacterial sepsis.


Amikacin Concentrations Predictive of Ototoxicity in Multidrug-Resistant Tuberculosis Patients.
Aminoglycosides, such as amikacin, are used to treat multidrug-resistant tuberculosis. However, ototoxicity is a common problem and is monitored using peak and trough amikacin concentrations based on World Health Organization recommendations. Our objective was to identify clinical factors predictive of ototoxicity using an agnostic machine learning method. We used classification and regression tree (CART) analyses to identify clinical factors, including amikacin concentration thresholds that predicted audiometry-confirmed ototoxicity among 28 multidrug-resistant pulmonary tuberculosis patients in Botswana. Amikacin concentrations were measured for all patients. The quantitative relationship between predictive factors and the probability of ototoxicity were then identified using probit analyses. The primary predictors of ototoxicity on CART analyses were cumulative days of therapy, followed by cumulative area under the concentration-time curve (AUC), which improved on the primary predictor by 87%. The area under the receiver operating curve was 0.97 on the test set. Peak and trough were not predictors in any tree. When algorithms were forced to pick peak and trough as primary predictors, the area under the receiver operating curve fell to 0.46. Probit analysis revealed that the probability of ototoxicity increased sharply starting after 6 months of therapy to near maximum at 9 months. A 10% probability of ototoxicity occurred with a threshold cumulative AUC of 87,232 days · mg · h/liter, while that of 20% occurred at 120,000 days · mg · h/liter. Thus, cumulative amikacin AUC and duration of therapy, and not peak and trough concentrations, should be used as the primary decision-making parameters to minimize the likelihood of ototoxicity in multidrug-resistant tuberculosis.


Mechanisms by which dopamine receptors may influence synaptic plasticity.
While dopamine (DA) receptors mediate acute effects of amphetamine and cocaine, chronic drug administration produces many glutamate-dependent adaptations, including LTP in reward-related neuronal circuits. An important question presents itself: How do DA receptors influence glutamate-dependent synaptic plasticity? Alterations in AMPA receptor phosphorylation and trafficking are critical for LTP. We hypothesize that D1 DA receptors modulate these processes, that chronic drug-induced adaptations in D1 receptor signaling, therefore, trigger compensatory changes in AMPA receptor function, and that this ultimately contributes to inappropriate plasticity in addiction-related neuronal circuits. Postnatal rat nucleus accumbens (NAc) cultures were used to study D1 receptor regulation of the AMPA receptor subunit GluR1. We found that D1 receptor stimulation enhances phosphorylation of GluR1 at the protein kinase A (PKA) site. Furthermore, D1 receptor stimulation increases GluR1 surface expression by increasing the rate of GluR1 externalization. The latter effect is prevented by the PKA inhibitors KT5720 and RpcAMPS, whereas the PKA activator SpcAMPS increases the rate of GluR1 externalization. These findings indicate that PKA phosphorylation is important in determining AMPA receptor surface expression and suggest a mechanism by which DA-releasing drugs of abuse may directly tap into fundamental mechanisms that enable synaptic plasticity. A limitation of our current model is that there are no intrinsic glutamate neurons in the NAc and thus no glutamate synapses in NAc cultures. To address this problem, we have restored excitatory synaptic inputs to NAc neurons by co-culturing them with prefrontal cortex (PFC) neurons. We are also studying GluR1 trafficking in PFC cultures. In both systems, synaptic AMPA receptors can be defined based on colocalization of GluR1 and the synaptic marker synaptobrevin. Preliminary results suggest that D1 receptor stimulation or PKA activation leads to increased surface GluR1 expression in PFC neurons but not to insertion into synaptic sites.


Increasing the use of e-consultation in primary care: results of an online survey among non-users of e-consultation.
To identify factors that can enhance the use of e-consultation in primary care. We investigated the barriers, demands and motivations regarding e-consultation among patients with no e-consultation experience (non-users). We used an online survey to gather data. Via online banners on 26 different websites of patient organizations we recruited primary care patients with chronic complaints, an important target group for e-consultation. A regression analysis was performed to identify the main drivers for e-consultation use among patients with no e-consultation experience. In total, 1706 patients started to fill out the survey. Of these patients 90% had no prior e-consultation experience. The most prominent reasons for non-use of e-consultation use were: not being aware of the existence of the service, the preference to see a doctor and e-consultation not being provided by a GP. Patients were motivated to use e-consultation, because e-consultation makes it possible to contact a GP at any time and because it enabled patients to ask additional questions after a visit to the doctor. The use of a Web-based triage application for computer-generated advice was popular among patients desiring to determine the need to see a doctor and for purposes of self-care. The patients' motivations to use e-consultation strongly depended on demands being satisfied such as getting a quick response. When looking at socio-demographic and health-related characteristics it turned out that certain patient groups - the elderly, the less-educated individuals, the chronic medication users and the frequent GP visitors - were more motivated than other patient groups to use e-consultation services, but were also more demanding. The less-educated patients, for example, more strongly demanded instructions regarding e-consultation use than the highly educated patients. In order to foster the use of e-consultation in primary care both GPs and non-users must be informed about the possibilities and consequences of e-consultation through tailored education and instruction. We must also take into account patient profiles and their specific demands regarding e-consultation. Special attention should be paid to patients who can benefit the most from e-consultation while also facing the greatest chance of being excluded from the service. As health care continues to evolve towards a more patient-centred approach, we expect that patient expectations and demands will be a major force in driving the adoption of e-consultation.


Abuse Is Abuse: The Influence of Type of Abuse, Victim Age, and Defendant Age on Juror Decision Making.
The purpose of the current study was to examine the role of victim age, defendant age, and type of abuse on mock juror decision making. Mock jurors (N = 556) read a trial transcript in which a soccer coach was accused of sexual abuse or physical abuse against a player. The victim's age (child, adolescent, or young adult), the defendant's age (young, middle age, or older adult), and the type of abuse (sexual or physical) were varied. Mock jurors provided a dichotomous and continuous verdict and rated their perceptions of the victim and the defendant. Although no differences on mock jurors' dichotomous verdict were found due to victim age, defendant age, or type of abuse, mock jurors provided higher guilt ratings when the abuse was sexual and both the victim and defendant were described as young adults. Similarly, mock jurors rated the victim more positively when the victim was described as a young adult (vs. child) for both sexual and physical abuse cases, and rated the defendant more positively when the victim was described as a child compared with young adult in sexual abuse cases. These findings suggest that mock jurors were largely influenced by victim age, particularly when the victim was described as an adult compared with a child.


Use of a fluorescent brightener to demonstrate cellulose in the cellular slime molds.
The presence and location of cellulose in different stages of the life cycles of the cellular slime molds can be demonstrated by use of the disodium salt of 4,4'-bis(4-anilino-6-bis (2-hydroxyethyl)-amino-s-triazin-2-ylamino)-2,2' -stilbene disulfonic acid, a fluorescent brightener. It may be used successfully as a direct stain at a concentration of 0.1% in half-normal saline at pH 6; and it may be incorporated into growth media as a vital stain at a concentration of 0.0025% with no inhibitory effect at any developmental stage. Vegetative myxamoebae contain no cellulose and show no fluorescence in the presence of this brightener when viewed with ultraviolet light. In later stages of the life cycle, the time and sites of cellulose formation can be demonstrated with the brightener because of its fluorescence. e.g., in the slime covering of the pseudoplasmodia, in the sorophore sheath, in the walls of stalk cells and spores, in the walls of microcysts, and in the walls and sheath material of macrocysts. The brightener appears to be a very sensitive indicator for cellulose, and it has certain advantages over other cellulose stains, since the staining reaction (fluorescence) is very intense, long-lasting, and not obscured by unstained cellulose-free myxamoebae if such are present.


Differentiation and maturation of embryonal carcinoma-derived neurons in cell culture.
We have previously shown that retinoic acid-treated cultures of the P19 line of embryonal carcinoma cells differentiate into neurons, glia, and fibroblast-like cells (Jones-Villeneuve et al., 1982). We report here that the monoclonal antibody HNK-1 reacts with the neurons at a very early stage of their differentiation and is, therefore, an early marker of the neuronal lineage. Cells in differentiated P 19 cultures synthesized acetylcholine but not catecholamines, suggesting that at least some of the neurons are cholinergic. The neurons also carry high-affinity uptake sites for GABA but not for serotonin. In long-term cultures, neuronal processes differentiated into axons and dendrites, which formed synapses. This biological system should prove valuable for examining the development and maturation of cholinergic neurons, since their differentiation occurs in cell culture.


α-Diazo oxime ethers for N-heterocycle synthesis.
This Feature Article introduces the preparation and synthetic utility of α-diazo oxime ethers. α-Oximino carbenes are useful synthons for N-heterocycles, and can be easily prepared from α-diazo oxime ethers as precursors. We begin with the preparation of α-diazo oxime ethers and their application in [3+2] cycloaddition. It turns out that the nature of metals bound to carbenes plays a crucial role in modulating the reactivity of α-oximino carbenes, in which copper carbenes smoothly react with enamines, whereas the less reactive enol ethers and nitriles require gold carbenes. In Section 3.2, a discussion on N-O and C-H bond activation is presented. Carbenes derived from diazo oxime ethers show unique reactivity towards N-O and C-H bond activation, in which the proximity of the two functionalities, carbene and oxime ether, dictates the preferred reaction pathways toward pyridines, pyrroles, and 2H-azirines. In Section 3.3, the development of tandem reactions based on α-diazo oxime ethers is discussed. The nature of carbenes in which whether free carbenes or metal complexes are involved dissects the pathway and forms different types of 2H-azirines. The 2H-azirine formation turned out to be an excellent platform for the tandem synthesis of N-heterocycles including pyrroles and pyridines. In the last section, we describe the electrophilic activation of 2H-azirines with vinyl carbenes and oximino carbenes. The resulting azirinium species undergo rapid ring expansion rearrangements to form pyridines and pyrazines.


Influence of halogen substitutions on rates of charge tunneling across SAM-based large-area junctions.
This paper examines the ability of structural modifications using halogen atoms (F, Cl, Br, and I) to influence tunneling rates across self-assembled monolayer (SAM)-based junctions having the structure Ag(TS)/S(CH2)n(p-C6H4X)//Ga2O3/EGaIn, where S(CH2)n(p-C6H4X) is a SAM of benzenethiol (n = 0) or benzyl mercaptan (n = 1) terminated in a hydrogen (X = H) or a halogen (X = F, Cl, Br, or I) at the para-position. The measured tunneling current densities (J(V); A cm(-2)) indicate that replacing a terminal hydrogen with a halogen atom at the X//Ga2O3 interface leads to a decrease in J(V) by ∼×13 for S(p-C6H4X) and by ∼×50 for SCH2(p-C6H4X). Values of J(V) for the series of halogenated SAMs were indistinguishable, indicating that changes in dipole moment and polarizability caused by introducing different halogen atoms at the interface between the SAM and the Ga2O3/EGaIn electrode do not significantly influence the rates of charge tunneling across the junctions.


Model-based analysis of multiple electrode array stimulation for epiretinal visual prostheses.
Epiretinal stimulation, which uses an array of electrodes implanted on the inner retinal surface to relay a representation of the visual scene to the neuronal elements of the retina, has seen considerable success. The objective of the present study was to quantify the effects of multi-electrode stimulation on the patterns of neural excitation in a computational model of epiretinal stimulation. A computational model of retinal ganglion cells was modified to represent the morphology of human retinal ganglion cells and validated against published experimental data. The ganglion cell model was then combined with a model of an axon of the nerve fiber layer to produce a population model of the inner retina. The response of the population of model neurons to epiretinal stimulation with a multi-electrode array was quantified across a range of electrode geometries using a novel means to quantify the model response-the minimum radius circle bounding the activated model neurons as a proxy for the evoked phosphene. Multi-electrode stimulation created unique phosphenes, uch that the number of potential phosphenes can far exceed the number of electrode contacts. The ability to exploit the spatial and temporal interactions of stimulation may be critical to improvements in the performance of epiretinal prostheses.


Hypogonadism predisposes males to the development of behavioural and neuroplastic depressive phenotypes.
The incidence of depression is 2-3× higher in women particularly during the reproductive years, an occurrence that has been associated with levels of sex hormones. The age-related decline of testosterone levels in men corresponds with the increased acquisition of depressive symptoms, and hormone replacement therapy can be efficacious in treating depression in hypogonadal men. Although it is not possible to model depression in rodents, it is possible to model some of the symptoms of depression including a dysregulated stress response and altered neuroplasticity. Among animal models of depression, chronic mild unpredictable stress (CMS) is a common paradigm used to induce depressive-like behaviours in rodents, disrupt the hypothalamic-pituitary adrenal axis and decrease hippocampal neuroplasticity. The purpose of this study was to assess the effect of hypogonadism, produced by gonadectomy, on the acquisition of depressive-like behaviours and changes in hippocampal neuroplasticity in adult male Sprague-Dawley rats. A 21-day unpredictable CMS protocol was used on gonadectomised (GDX) and sham-operated males which produced an attenuation of weight gain in the GDX males receiving CMS treatment (GDX-CMS). Behavioural analysis was carried out to assess anxiety- and depressive-like behaviours. The combination of GDX and CMS produced greater passive behaviours within the forced swim test than CMS exposure alone. Similarly, hippocampal cell proliferation, neurogenesis and the expression of the neuroplastic protein polysialated neural cell adhesion molecule (PSA-NCAM) were all significantly reduced in the GDX-CMS group compared to all other treatment groups. These findings indicate that testicular hormones confer resiliency to chronic stress in males therefore reducing the likelihood of developing putative physiological, behavioural or neurological depressive-like phenotypes.


C-terminal HERG (LQT2) mutations disrupt IKr channel regulation through 14-3-3epsilon.
Beta-adrenergic receptor-mediated cAMP or protein kinase A (PKA)-dependent modulation of cardiac potassium currents controls ventricular action potential duration (APD) at faster heart rates. HERG (KCNH2) gene mutations are associated with congenital long-QT syndrome (LQT2) and affect IKr activity, a key determinant in ventricular repolarization. Physical activity or emotional stress often triggers lethal arrhythmias in LQT2 patients. Beta-adrenergic stimulation of HERG channel activity is amplified and prolonged in vitro by the adaptor protein 14-3-3epsilon. In LQT2 families, we identified three novel heterozygous HERG mutations (G965X, R1014PfsX39, V1038AfsX21) in the C-terminus that led to protein truncation and loss of a PKA phosphorylation site required for binding of 14-3-3epsilon. When expressed in CHO cells, the mutants produced functional HERG channels with normal kinetic properties. We now provide evidence that HERG channel regulation by 14-3-3epsilon is of physiological significance in humans. Upon co-expression with 14-3-3epsilon, mutant channels still bound 14-3-3epsilon but did not respond with a hyperpolarizing shift in voltage dependence as seen in wild-type channels. Co-expression experiments of wild-type and mutant channels revealed dominant-negative behavior of all three HERG mutations. Simulations of the effects of sympathetic stimulation of HERG channel activity on the whole-cell action potential suggested a role in rate-dependent control of APD and an impaired ability of mutant cardiac myocytes to respond to a triggered event or an ectopic beat. In summary, the attenuated functional effects of 14-3-3epsilon on C-terminally truncated HERG channels demonstrate the physiological importance of coupling beta-adrenergic stimulation and HERG channel activity.


Utilization of Bt corn residues by grazing beef steers and Bt corn silage and grain by growing beef cattle and lactating dairy cows.
Three experiments were conducted to evaluate the impact of the Bacillus thuringiensis (Bt)-11 transformation event in two parental corn hybrids differing in date of maturity on beef and dairy cattle performance. Sixteen lactating Holstein dairy cows in replicated 4 x 4 Latin squares were assigned to four diets in a 2 x 2 factorial arrangement: Bt vs non-Bt trait and early- vs late-maturing corn hybrids. The diets contained 40% of the test corn silage plus 28% corn grain from the same corn hybrid (DM basis). There was no effect of the Bt trait on efficiency of milk production, ruminal pH, acetate:propionate ratio, or in situ digestion kinetics of NDF. The early-maturing corn hybrids resulted in greater total VFA concentrations in the rumen and efficiency of 4% fat-corrected milk production than the later-maturing hybrids (P < 0.05). Sixty-seven steer calves were used in a 70-d corn residue grazing trial for the late-maturing corn hybrids only. Daily BW gain of steers was similar for those grazing Bt and non-Bt corn residues, and the steers exhibited no grazing preference between Bt and non-Bt corn residue. One hundred twenty-eight steer calves were assigned to four silage-based growing diets in a 2 x 2 factorial arrangement: Bt vs non-Bt trait and early- vs late-maturing corn hybrids. The diets contained 90% corn silage and 10% supplement (DM basis). The DMI was higher for steers fed Bt compared with non-Bt hybrids (P < 0.02). An interaction (P < 0.03) was observed for feed efficiency between hybrid genotype and incorporation of the Bt trait. Feed efficiency was greater (P < 0.05) for steers fed the later-maturing non-Bt hybrid compared with the later-maturing Bt hybrid; however, feed efficiency was similar between steers fed early-maturing Bt and non-Bt corn silages. Steers fed the early-maturing hybrid gained 11% faster and were 7% more efficient compared with those fed the late-maturing hybrid. These latter results agree with the dairy experiment in which the early-maturing hybrid resulted in 5% greater efficiency of milk production than the later-maturing corn hybrid. In all experiments, incorporation of the Bt trait into corn had no consistent effect on cattle performance. In addition, background genetics of the corn hybrids appeared to have a more consistent impact on performance than did presence of the Bt trait.


Expression profiling of archival renal tumors by quantitative PCR to validate prognostic markers.
Formalin-fixed paraffin-embedded (FFPE) tissues are routinely stored by most pathology departments and are a widely available resource for discovery of clinically useful biomarkers. We describe our method for optimizing quantitative reverse transcription PCR (RT-PCR) for expression analysis using frozen and archival tissue. Commonly used reference genes were evaluated for stability of expression in normal kidney and clear cell renal cell carcinoma (RCC). Optimal reference genes for calculating normalization factors for RT-PCR were ACTB, RPL13A, GUS, RPLP0, HPRT1, and SDHA when using FFPE RCC. The optimal reference genes when using frozen RCC were ACTB, RPL13A, and GUS, confirming that use of multiple reference genes improves accuracy when intact RNA from frozen renal tumors are used. Expression of 16 markers previously reported to have prognostic significance in RCC was determined in 23 matching frozen and FFPE renal tumors, representing a range of tumor grades and stages; correlation coefficient for expression measured in frozen and FFPE tumors was 0.921 (P < 0.001). All markers predicted survival when frozen tumors were used and 14 of the 16 markers predicted survival when FFPE tumors were used as the source of RNA. An optimized RT-PCR assay can accurately measure expression of most prognostic tumor markers.