Steric exclusion and constraint satisfaction in multi-scale coarse-grained simulations.
An algorithm is described for the interaction of a hierarchy of objects that seeks to circumvent a fundamental problem in coarse-grained modelling which is the loss of fine detail when components become bundled together. A "currants-in-jelly" model is developed that provides a flexible approach in which the contribution of the soft high-level objects (jelly-like) are employed to protect the underlying atomic structure (currants), while still allowing them to interact. Idealised chains were used to establish the parameters to achieve this degree of interaction over a hierarchy spanning four levels and in a more realistic example, the distortion experienced by a protein domain structure during collision was measured and the parameters refined. This model of steric repulsion was then combined with sets of predicted distance constraints, derived from correlated mutation analysis. Firstly, an integral trans-membrane protein was modelled in which the packing of the seven helices was refined but without topological rearrangement. Secondly, an RNA structure was 'folded' under the predicted constraints, starting only from its 2-dimensional secondary structure prediction.


Lipoprotein-associated phospholipase A2 as an independent predictor of coronary heart disease. West of Scotland Coronary Prevention Study Group.
Chronic inflammation is believed to increase the risk of coronary events by making atherosclerotic plaques in coronary vessels prone to rupture. We examined blood constituents potentially affected by inflammation as predictors of risk in men with hypercholesterolemia who were enrolled in the West of Scotland Coronary Prevention Study, a trial that evaluated the value of pravastatin in the prevention of coronary events. A total of 580 men who had had a coronary event (nonfatal myocardial infarction, death from coronary heart disease, or a revascularization procedure) were each matched for age and smoking status with 2 control subjects (total, 1160) from the same cohort who had not had a coronary event. Lipoprotein-associated phospholipase A2, C-reactive protein, and fibrinogen levels, and the white-cell count were measured at base line, along with other traditional risk factors. The association of these variables with the risk of coronary events was tested in regression models and by dividing the range of values according to quintiles. Levels of C-reactive protein, the white-cell count, and fibrinogen levels were strong predictors of the risk of coronary events; the risk in the highest quintile of the study cohort for each variable was approximately twice that in the lowest quintile. However, the association of these variables with risk was markedly attenuated when age, systolic blood pressure, and lipoprotein levels were included in multivariate models. Levels of lipoprotein-associated phospholipase A2 (platelet-activating factor acetylhydrolase), the expression of which is regulated by mediators of inflammation, had a strong, positive association with risk that was not confounded by other factors. It was associated with almost a doubling of the risk in the highest quintile as compared with the lowest quintile. Inflammatory markers are predictors of the risk of coronary events, but their predictive ability is attenuated by associations with other coronary risk factors. Elevated levels of lipoprotein-associated phospholipase A2 appear to be a strong risk factor for coronary heart disease, a finding that has implications for atherogenesis and the assessment of risk.


Application of Accelerated Time Models to Compare Performance of Two Comorbidity-adjusting Methods with APACHE II in Predicting Short-term Mortality Among the Critically Ill.
This study aimed to determine how the abilities of the Charlson Index and Elixhauser comorbidities compared with the chronic health components of the Acute Physiology and Chronic Health Evaluation (APACHE II) to predict in-hospital 30 day mortality among adult critically ill patients treated inside and outside of Intensive Care Unit (ICU). A total of 701 critically ill patients, identified in a prevalence study design on four randomly selected days in five acute care hospitals, were followed up from the date of becoming critically ill for 30 days or until death, whichever occurred first. Multiple data sources including administrative, clinical, pathology, microbiology and laboratory patient records captured the presence of acute and chronic illnesses. The exponential, Gompertz, Weibull, and log-logistic distributions were assessed as candidate parametric distributions available for the modelling of survival data. Of these, the log-logistic distribution provided the best fit and was used to construct a series of parametric survival models. Of the 701 patients identified in the initial prevalence study, 637 (90.9%) had complete data for all fields used to calculate APACHE II score. Controlling for age, sex and Acute Physiology Score (APS), the chronic health components of the APACHE II score, as a group, were better predictors of survival than Elixhauser comorbidities and Charlson Index. Of the APACHE II chronic health components, only the relatively uncommon conditions of liver failure (3.4%) and immunodeficiency (9.6%) were statistically associated with inferior patient survival with acceleration factors of 0.35 (95% CI 0.17, 0.72) for liver failure, and 0.42 (95% CI 0.26, 0.72) for immunodeficiency. Sensitivity analyses on an imputed dataset that also included the 64 individuals with imputed APACHE II score showed identical results. Our study suggests that, in acute critical illness, most co-existing comorbidities are not major determinants of shortterm survival, indicating that observed variations in ICU patient 30-day mortality may not be confounded by lack of adjustment to pre-existing comorbidities.


Lesser White-fronted (Anser erythropus) and Greater White-fronted (A. albifrons) Geese wintering in Greek wetlands are not threatened by Pb through shot ingestion.
Fecal lead (Pb) levels were investigated in the threatened European population of the Lesser White-fronted Goose (LWfG, Anser erythropus) and of the non-threatened Greater White-fronted Goose (GWfG, Anser albifrons) wintering in two wetland areas in northern Greece in order to assess the potential risk from Pb exposure. Fecal, soil and food plant samples were analyzed. Levels of Pb were normalized using Al concentrations in order to separate the effect of possible ingestion of Pb shot from that of soil or sediment accidentally ingested with food. All concentrations are expressed on a dry weight basis. Geometric means of Pb content in the feces of LWfG were 6.24 mg/kg at Evros Delta and 7.34 mg/kg at Lake Kerkini (maximum values of 28.61 mg/kg and 36.68 mg/kg, respectively); for fecal samples of GWfG geometric means were 2.39 mg/kg at Evros Delta and 6.90 mg/kg at Kerkini (corresponding maximum values of 25.09 mg/kg and 42.26 mg/kg). Soil Pb was in the range of 5.2-60.2mg/kg (geometric mean = 22.6 mg/kg) for the Evros Delta and between 13.4 and 64.9 mg/kg (geometric mean=28.1mg/kg) for Kerkini. A general linear model fitted to the data showed that Pb levels were very closely dependent on Al levels in the feces from both species and at both sites indicating soil or sediment were the only significant source of Pb; species and site, as well as their interaction, were not statistically significant factors. For both species and at both sites exposure to Pb was evidently very mild and the observed levels of Pb were well below the proposed thresholds for lethal or sublethal effects of Pb poisoning. Soil ingestion appeared to gradually increase from October to December for LWfG at Kerkini, corresponding to a gradual depletion of their food source.


Domestic trends in malaria research and development in China and its global influence.
Though many countries, including China, are moving towards malaria elimination, malaria remains a major global health threat. Due to the spread of antimalarial drug resistance and the need for innovative medical products during the elimination phase, further research and development (R&D) of innovative tools in both epidemic and elimination areas is needed. This study aims to identify the trends and gaps in malaria R&D in China, and aims to offer suggestions on how China can be more effectively involved in global malaria R&D. Quantitative analysis was carried out by collecting data on Chinese malaria-related research programmes between 1985 and 2014, invention patents in China from 1985 to 2014, and articles published by Chinese researchers in PubMed and Chinese databases from 2005 to 2014. All data were screened and extracted for numerical analysis and were categorized into basic sciences, drug/drug resistance, immunology/vaccines, or diagnostics/detection for chronological and subgroup comparisons. The number of malaria R&D activities have shown a trend of increase during the past 30 years, however these activities have fluctuated within the past few years. During the past 10 years, R&D on drug/drug resistance accounted for the highest percentages of research programmes (32.4%), articles (55.0% in PubMed and 50.6% in Chinese databases) and patents (45.5%). However, these R&D activities were mainly related to artemisinin. R&D on immunology/vaccines has been a continuous interest for China's public entities, but the focus remains on basic science. R&D in the area of high-efficiency diagnostics has been rarely seen or reported in China. China has long been devoted to malaria R&D in multiple areas, including drugs, drug resistance, immunology and vaccines. R&D on diagnostics has received significantly less attention, however, it should also be an area where China can make a contribution. More focus on malaria R&D is needed, especially in the area of diagnostics, if China would like to contribute in a more significant way to global malaria control and elimination.


Improved outcome of N-butyldeoxygalactonojirimycin-mediated substrate reduction therapy in a mouse model of Sandhoff disease.
Sandhoff disease is a severe neurodegenerative glycosphingolipid (GSL) lysosomal storage disorder, currently without treatment options. One therapeutic approach under investigation is substrate reduction therapy (SRT). By partially inhibiting GSL biosynthesis, the impaired rate of GSL catabolism is balanced by a slower rate of influx of GSLs into the lysosome. In a previous study, we reported the beneficial effects of treating Sandhoff disease mice with the glucose analogue N-butyldeoxynojirimycin (NB-DNJ), a compound that inhibits the first step of GSL biosynthesis catalysed by the ceramide specific glucosyltransferase. NB-DNJ, however, exhibits adverse effects at high doses such as weight loss and GI tract distress (due to glucosidase inhibition). This might limit the therapeutic potential of NB-DNJ for treating diseases affecting the CNS where high dose therapy may be required to achieve therapeutic levels of the drug in the brain. In the present study, a more selective compound, the galactose analogue N-butyldeoxygalactonojirimycin (NB-DGJ), was evaluated in the Sandhoff disease mouse model. Treatment with NB-DGJ showed greater therapeutic efficacy than NB-DNJ with no detectable side effects. The ability to escalate the dose of NB-DGJ, leading to extended life expectancy and increased delay in symptom onset, demonstrates the greater therapeutic potential of NB-DGJ for the treatment of the human gangliosidoses.


Measure the Semantic Similarity of GO Terms Using Aggregate Information Content.
The rapid development of gene ontology (GO) and huge amount of biomedical data annotated by GO terms necessitate computation of semantic similarity of GO terms and, in turn, measurement of functional similarity of genes based on their annotations. In this paper we propose a novel and efficient method to measure the semantic similarity of GO terms. The proposed method addresses the limitations in existing GO term similarity measurement techniques; it computes the semantic content of a GO term by considering the information content of all of its ancestor terms in the graph. The aggregate information content (AIC) of all ancestor terms of a GO term implicitly reflects the GO term's location in the GO graph and also represents how human beings use this GO term and all its ancestor terms to annotate genes. We show that semantic similarity of GO terms obtained by our method closely matches the human perception. Extensive experimental studies show that this novel method also outperforms all existing methods in terms of the correlation with gene expression data. We have developed web services for measuring semantic similarity of GO terms and functional similarity of genes using the proposed AIC method and other popular methods. These web services are available at http://bioinformatics.clemson.edu/G-SESAME.


Laser microdissection microscopy and single cell PCR of avian hemosporidians.
Avian malaria parasites (Plasmodium spp.) and related species of Haemoproteus constitute a remarkably diverse and species rich group of blood parasites. Analyses of the mitochondrial cytochrome b gene of these hemosporidians have demonstrated unexpected patterns of host distribution, host shifts, and host sharing. However, deeper insights into these patterns require access to multiple genetic markers and genetic analyses of single parasite cells. In the present study, we demonstrate the potential of laser microdissection microscopy (Olympus/MMI CellCut microdissection system) for solving these 2 problems. This technique was used for isolation of single blood stages and ookinetes of avian Haemoproteus and Plasmodium spp., which were then successfully used for DNA isolation, amplification, and sequencing. The methods of single cell dissection of hemosporidian parasites and PCR-based analyses with dissected single cells are described. These methods can be used to isolate substantial quantities of pure hemosporidian parasite DNA for large-scale sequencing, essential information when designing primers for developing multiple nuclear genetic markers. Such markers can then be applied to isolated single parasite cells for identification of parasites in mixed infections and deciphering mechanisms behind apparent reproductive isolation between parasite lineages. This method can be used in the molecular investigation of blood parasites of birds, reptiles, and fish because it enables removing the parasite DNA from the overpowering host DNA, which is present in red blood cells.


Delivery of genes into the CF airway.
Gene therapy was suggested as a potential treatment for cystic fibrosis (CF), even before the identification of the CFTR gene. Initial enthusiasm has been tempered as it became apparent that reintroduction of the CFTR gene into the cells of the lung is more difficult than anticipated. Here, we review the major gene delivery vectors evaluated clinically, and suggest that advances in either plasmid DNA design and/or hybrid lentivirus biology may finally facilitate lung gene transfer with efficiencies sufficient for CF gene therapy to offer clinical benefit.


Increased condom use at sexual debut in the general population of Slovenia and association with subsequent condom use.
To describe the extent of condom use at first heterosexual intercourse (FHI) in the general population of Slovenia, identify associated factors and assess the association with subsequent use. Cross-sectional survey comprising the first Slovenian National Survey of Sexual Lifestyles, Attitudes and Health. Data were collected during 1999-2001 from a probability sample of the general population aged 18-49 years. Statistical methods for complex survey data were used to carry out weighted analyses. Condom use at FHI was reported by 23.6% of men and 21.3% of women. The more recently they experienced FHI, the more likely respondents were to report condom use (71.7% of men; 63.8% of women with FHI during 1995-1999). Men with FHI at age 18 or older and those with higher levels of education were more likely to use a condom and those who lost control (because drunk or carried away by feelings) less likely. Men seem to have more control over condom use at FHI than women. Sexually active men and women who used condom at FHI were 11 and 2.5 times more likely to consistently use condoms during the month preceding the interview. Currently, the majority of Slovenian men and women use protection against HIV, other sexually transmitted infections, and unplanned conception at FHI. The steep increase over time in condom use at FHI suggests that HIV-related condom use promotion has had an impact on preventive behaviours and should be sustained, especially since condom use at FHI predicts subsequent use.


Risk factors and adverse perinatal outcomes among term and preterm infants born small-for-gestational-age: secondary analyses of the WHO Multi-Country Survey on Maternal and Newborn Health.
Small for gestational age (SGA) is not only a major indicator of perinatal mortality and morbidity, but also the morbidity risks in later in life. We aim to estimate the association between the birth of SGA infants and the risk factors and adverse perinatal outcomes among twenty-nine countries in Africa, Latin America, the Middle East and Asia in 359 health facilities in 2010-11. We analysed facility-based, cross-sectional data from the WHO Multi-country Survey on Maternal and Newborn Health. We constructed multilevel logistic regression models with random effects for facilities and countries to estimate the risk factors for SGA infants using country-specific birthweight reference standards in preterm and term delivery, and SGA's association with adverse perinatal outcomes. We compared the risks and adverse perinatal outcomes with appropriate for gestational age (AGA) infants categorized by preterm and term delivery. A total of 295,829 singleton infants delivered were analysed. The overall prevalence of SGA was highest in Cambodia (18.8%), Nepal (17.9%), the Occupied Palestinian Territory (16.1%), and Japan (16.0%), while the lowest was observed in Afghanistan (4.8%), Uganda (6.6%) and Thailand (9.7%). The risk of preterm SGA infants was significantly higher among nulliparous mothers and mothers with chronic hypertension and preeclampsia/eclampsia (aOR: 2.89; 95% CI: 2.55-3.28) compared with AGA infants. Higher risks of term SGA were observed among sociodemographic factors and women with preeclampsia/eclampsia, anaemia and other medical conditions. Multiparity (> = 3) (AOR: 0.88; 95% CI: 0.83-0.92) was a protective factor for term SGA. The risk of perinatal mortality was significantly higher in preterm SGA deliveries in low to high HDI countries. Preterm SGA is associated with medical conditions related to preeclampsia, but not with sociodemographic status. Term SGA is associated with sociodemographic status and various medical conditions.


Use of ultrasound scanning and body condition score to evaluate composition traits in mature beef cows.
The experiment was designed to validate the use of ultrasound to evaluate body composition in mature beef cows. Both precision and accuracy of measurement were assessed. Cull cows (n = 87) selected for highly variable fatness were used. Two experienced ultrasound technicians scanned and assigned BCS to each cow on 2 consecutive days. Ultrasound traits were backfat thickness (UBFT), LM area (ULMA), body wall thickness (UBWT), rump fat depth (URFD), rump muscle depth (URMD), and intramuscular fat (UIMF; %). Cows were then harvested. Carcass traits were HCW, backfat thickness (CBFT), LM area (CLMA), body wall thickness (CBWT), and marbling score (CMS). Correlations between consecutive live measurements were greatest for subcutaneous fat (r > 0.94) and lower for BCS (r > 0.74) and URMD (r > 0.66). Repeatability bias differed from 0 for only 1 technician for URMD and UIMF (P < 0.01). Technicians differed in repeatability SE for only ULMA (P < 0.05). Correlations between live and carcass measurements were high for backfat and body wall thickness (r > 0.90) and slightly less for intramuscular fat and LM area (r = 0.74 to 0.79). Both technicians underestimated all carcass traits with ultrasound, but only CBFT and CBWT prediction bias differed from 0 (P < 0.05). Technicians had similar prediction SE for all traits (P > 0.05). Technician effects generally explained <1% of the total variation in precision. After accounting for technician, animal effects explained 50.4% of remaining variation in differences between repeated BCS (P < 0.0001) but were minimal for scan differences. When cows with mean BCS <4 or >7 were removed, the portion of remaining variation between repeated measurements defined by animal effects increased for most traits and was significant for UBFT and URFD (P = 0.03). Technician effects explained trivial variation in accuracy (P > 0.24). Animal effects explained 87.2, 75.2, and 81.7% (P < 0.0001) of variation remaining for CBFT, CLMA, and CBWT prediction error, respectively, and remained large and highly important (P < 0.0001) when only considering cows with BCS from 4 to 7. We conclude that experienced ultrasound technicians can precisely and accurately measure traits indicative of composition in mature beef cows. However, animal differences define substantial variation in scan differences and, especially, prediction errors. Implications for technician certification, carcass pricing, and genetic evaluation are discussed.


Transporting Children in Autonomous Vehicles: An Exploratory Study.
Identify factors that impact parents' decisions about allowing an unaccompanied child to ride in an autonomous vehicle (AV). AVs are being tested in several U.S. cities and on highways in multiple states. Meanwhile, suburban parents are using ridesharing services to shuttle children from school to extracurricular activities. Parents may soon be able to hire AVs to transport children. Nineteen parents of 8- to 16-year-old children, and some of their children, rode in a driving simulator in autonomous mode, then were interviewed. Parents also participated in focus groups. Topics included minimum age for solo child passengers, types of trips unaccompanied children might take, and vehicle features needed to support child passengers. Parents would require two-way audio communication and prefer video feeds of vehicle interiors, seatbelt checks, automatic locking, secure passenger identification, and remote access to vehicle information. Parents cited convenience as the greatest benefit and fear that AVs could not protect passengers during unplanned trip interruptions as their greatest concern. Manufacturers have an opportunity to design family-friendly AVs from the outset, rather than retrofit them to be safe for child passengers. More research, especially usability studies where families interact with technology prototypes, is needed to understand how AV design impacts child passengers. Potential applications of this research include not only designing vehicles that can be used to safely transport children, seniors who no longer drive, and individuals with disabilities but also developing regulations, policies, and societal infrastructure to support safe child transport via AVs.


Hybrid processes in enzymatically gelled gelatin: impact on , macroscopic properties and cellular response.
Physical, chemical and hybrid tilapia fish gelatin hydrogels were investigated by small-angle neutron scattering (), molecular dynamic simulations and their biological effect in cell cultures studied; results from the different experimental techniques were then correlated and linked to the rheological properties of the gels (F. Bode et al., Biomacromolecules, 2011, 12, 3741-3752). Hydrogels were obtained by cross-linking with the microbial enzyme transglutaminase (mTGase) under two conditions: above and below gelatin physical temperature (ca. 23 °C). Hydrogels cross-linked at 37 °C, from the sol-state, are referred to as 'chemical' gels (C); hydrogels cross-linked at 21 °C, thus with concurrent physical , are referred to as 'physical-co-chemical' gels (PC). The data were appropriately described by a combination of a Lorentzian and a power law model. For physical gels, the correlation length (ξ) obtained from the fits decreased linearly with gelatin concentration, from 42 to 26 Å for 3.5 to 10% w/w gelatin, respectively. Independently of temperature, all physical gels at a given concentration showed a similar correlation length ξ (26 ± 2 Å), with no significant difference with the sol-state (23 ± 2 Å). In both C and PC gels, ξ increased with mTGase concentration over the range studied: 40 to 167 Å for 10 and 40 U mTGase per g gelatin in C gels (after 120 min cross-linking) and 40 to 82 Å for 10 and 40 U mTGase per g gelatin for PC gels. ξ reached a plateau at the highest mTGase concentration studied for both types of gels. In addition, kinetic studies on C gels revealed that ξ increased linearly with time in the first two hours and grew faster with increasing mTGase concentration. ξ values in the PC gels were smaller than in the corresponding C gels. Cell proliferation studies showed that the gels were compatible with cell growth and indicated no statistically relevant dependence on mTGase concentration for C gels. For PC gels, cell proliferation decreased with increases in mTGase concentration, by approximately 80% from 10 to 40 U mTGase per g gelatin. With the exception of the highest mTGase concentration studied, PC gels overall showed a slightly (but statistically significant) higher cell proliferation than the corresponding chemical gels.


Drosophila E-cadherin is required for the maintenance of ring canals anchoring to mechanically withstand tissue growth.
Intercellular bridges called "ring canals" (RCs) resulting from incomplete cytokinesis play an essential role in intercellular communication in somatic and germinal tissues. During Drosophila oogenesis, RCs connect the maturing oocyte to nurse cells supporting its growth. Despite numerous genetic screens aimed at identifying genes involved in RC biogenesis and maturation, how RCs anchor to the plasma membrane (PM) throughout development remains unexplained. In this study, we report that the clathrin adaptor protein 1 (AP-1) complex, although dispensable for the biogenesis of RCs, is required for the maintenance of the anchorage of RCs to the PM to withstand the increased membrane tension associated with the exponential tissue growth at the onset of vitellogenesis. Here we unravel the mechanisms by which AP-1 enables the maintenance of RCs' anchoring to the PM during size expansion. We show that AP-1 regulates the localization of the intercellular adhesion molecule E-cadherin and that loss of AP-1 causes the disappearance of the E-cadherin-containing adhesive clusters surrounding the RCs. E-cadherin itself is shown to be required for the maintenance of the RCs' anchorage, a function previously unrecognized because of functional compensation by N-cadherin. Scanning block-face EM combined with transmission EM analyses reveals the presence of interdigitated, actin- and Moesin-positive, microvilli-like structures wrapping the RCs. Thus, by modulating E-cadherin trafficking, we show that the sustained E-cadherin-dependent adhesion organizes the microvilli meshwork and ensures the proper attachment of RCs to the PM, thereby counteracting the increasing membrane tension induced by exponential tissue growth.


Sex-specific demography and generalization of the Trivers-Willard theory.
The Trivers-Willard theory proposes that the sex ratio of offspring should vary with maternal condition when it has sex-specific influences on offspring fitness. In particular, mothers in good condition in polygynous and dimorphic species are predicted to produce an excess of sons, whereas mothers in poor condition should do the opposite. Despite the elegance of the theory, support for it has been limited. Here we extend and generalize the Trivers-Willard theory to explain the disparity between predictions and observations of offspring sex ratio. In polygynous species, males typically have higher mortality rates, different age-specific reproductive schedules and more risk-prone life history tactics than females; however, these differences are not currently incorporated into the Trivers-Willard theory. Using two-sex models parameterized with data from free-living mammal populations with contrasting levels of sex differences in demography, we demonstrate how sex differences in life history traits over the entire lifespan can lead to a wide range of sex allocation tactics, and show that correlations between maternal condition and offspring sex ratio alone are insufficient to conclude that mothers adaptively adjust offspring sex ratio.


Hitting a Moving Target: A Model for Malaria Elimination in the Presence of Population Movement.
South Africa is committed to eliminating malaria with a goal of zero local transmission by 2018. Malaria elimination strategies may be unsuccessful if they focus only on vector biology, and ignore the mobility patterns of humans, particularly where the majority of infections are imported. In the first study in Mpumalanga Province in South Africa designed for this purpose, a metapopulation model is developed to assess the impact of their proposed elimination-focused policy interventions. A stochastic, non-linear, ordinary-differential equation model is fitted to malaria data from Mpumalanga and neighbouring Maputo Province in Mozambique. Further scaling-up of vector control is predicted to lead to a minimal reduction in local infections, while mass drug administration and focal screening and treatment at the Mpumalanga-Maputo border are predicted to have only a short-lived impact. Source reduction in Maputo Province is predicted to generate large reductions in local infections through stemming imported infections. The mathematical model predicts malaria elimination to be possible only when imported infections are treated before entry or eliminated at the source suggesting that a regionally focused strategy appears needed, for achieving malaria elimination in Mpumalanga and South Africa.


The effects of omega 3 fatty acid supplementation on brain tissue oxidative status in aged wistar rats.
The omega 3 fatty acids play an important role in many physiological processes. Their effect is well documented in neurodegenerative diseases and inflammatory diseases. Also, aging as a biophysiological process could be influenced by eicosapentanoic acid (EPA) and docosahexanoic acid (DHA) components of fish oil. However there are not many studies showing the effect of PUFA (polyunsaturated FA) suplementation in eldery brain functions and the response to oxidative strees. The aim of this study was to investigate the effects of dietary omega-3 fatty acid supplementation on levels of lipid peroxidation and oxidant/antioxidant status of brain tissue in aged (24 months old) Wistar rats. Animals were divided in two groups. Control group (n=8) was fed with standard laboratory food and received water ad libitum. Treated group (n=8) was also fed with standard laboratory food, water ad libitum and received fish oil capsules (EPA+DHA) for 6 weeks. Daily dose was 30mg EPA and 45mg DHA (capsules: 200mg EPA and 300mg DHA; in-house method). At the end of treatment animals were sacrificed and brains were collected and frozen on -80ºC. The levels of lipid peroxidation (malondialdehyde - MDA), activity of catalase (CAT) and activity of superoxide dismutase (SOD) were examined in cerebral cortex. Catalase activity was determined by measuring the decrease in absorbance (H2O2 degradation) at 240 nm for 3 min and expressed as U/mg protein. Total SOD (superoxide dismutase) activity was performed at room temperature according to the method of Misra and Fridovich. The extent of lipid peroxidation (LPO) was estimated as the concentration of thiobarbituric acid reactive product malondialdehyde (MDA) by using the method of Aruoma et al. The incorporation of fatty acids in cellular membranes was confirmed by gas chromatography. Our results showed that lipid peroxidation significantly decreased in treated animal group, where MDA concentration was 0.38±0.001 vs. 0.43±0.001 nM/ml (p<0.05) in control. However SOD activity increased significantly in treated animal group 1.57±0.24 vs. 4.12±0.15 U/gHb/L (p<0.01) in control. CAT activity decreased in treated group but not significantly. Incorporation of omega-3 fatty acids after their supplementation had beneficial effects on brain tissue. Omega-3 fatty acids increased activity of SOD and decreased lipid peroxidation. Changes in oxidative/antioxidative balance are a result of EPA and DHA effects on lipids and enzymes of antioxidative system.


Tai chi chuan for the primary prevention of stroke in middle-aged and elderly adults: a systematic review.
Background. Stroke is a major healthcare problem with serious long-term disability and is one of the leading causes of death in the world. Prevention of stroke is considered an important strategy. Methods. Seven electronic databases were searched. Results. 36 eligible studies with a total of 2393 participants were identified. Primary outcome measures, TCC exercise combined with other intervention had a significant effect on decreasing the incidence of nonfatal stroke (n = 185, RR = 0.11, 95% CI 0.01 to 0.85, P = 0.03) and CCD (n = 125, RR = 0.33, 95% CI 0.11 to 0.96, P = 0.04). For the risk factors of stroke, pooled analysis demonstrated that TCC exercise was associated with lower body weight, BMI, FBG level, and decreasing SBP, DBP, plasma TC, and LDL-C level regardless of the intervention period less than half a year or more than one year and significantly raised HDL-C level in comparison to nonintervention. Compared with other treatments, TCC intervention on the basis of the same other treatments in patients with chronic disease also showed the beneficial effect on lowering blood pressure. Conclusion. The present systematic review indicates that TCC exercise is beneficially associated with the primary prevention of stroke in middle-aged and elderly adults by inversing the high risk factors of stroke.


HealthPartners adopts community business model to deepen focus on nonclinical factors of health outcomes.
Clinical care contributes only 20 percent to overall health outcomes, according to a population health model developed at the University of Wisconsin. Factors contributing to the remainder include lifestyle behaviors, the physical environment, and social and economic forces--all generally considered outside the realm of care. In 2010 Minnesota-based HealthPartners decided to target nonclinical community health factors as a formal part of its strategic business plan to improve public health in the Twin Cities area. The strategy included creating partnerships with businesses and institutions that are generally unaccustomed to working together or considering how their actions could help improve community health. This article describes efforts to promote healthy eating in schools, reduce the stigma of mental illness, improve end-of-life decision making, and strengthen an inner-city neighborhood. Although still in their early stages, the partnerships can serve as encouragement for organizations inside and outside health care that are considering undertaking similar efforts in their markets.


Beta-keto amphetamines: studies on the metabolism of the designer drug mephedrone and toxicological detection of mephedrone, butylone, and methylone in urine using gas chromatography-mass spectrometry.
In recent years, a new class of designer drugs has appeared on the drugs of abuse market in many countries, namely, the so-called beta-keto (bk) designer drugs such as mephedrone (bk-4-methylmethamphetamine), butylone (bk-MBDB), and methylone (bk-MDMA). The aim of the present study was to identify the metabolites of mephedrone in rat and human urine using GC-MS techniques and to include mephedrone, butylone, and methylone within the authors' systematic toxicological analysis (STA) procedure. Six phase I metabolites of mephedrone were detected in rat urine and seven in human urine suggesting the following metabolic steps: N-demethylation to the primary amine, reduction of the keto moiety to the respective alcohol, and oxidation of the tolyl moiety to the corresponding alcohols and carboxylic acid. The STA procedure allowed the detection of mephedrone, butylone, methylone, and their metabolites in urine of rats treated with doses corresponding to those reported for abuse of amphetamines. Besides macro-based data evaluation, an automated evaluation using the automated mass spectral deconvolution and identification system was performed. Mephedrone and butylone could be detected also in human urine samples submitted for drug testing. Assuming similar kinetics in humans, the described STA procedure should be suitable for proof of an intake of the bk-designer drugs in human urine.


Hotmap: looking at geographic attention.
Understanding how people use online maps allows data acquisition teams to concentrate their efforts on the portions of the map that are most seen by users. Online maps represent vast databases, and so it is insufficient to simply look at a list of the most-accessed URLs. Hotmap takes advantage of the design of a mapping system's imagery pyramid to superpose a heatmap of the log files over the original maps. Users' behavior within the system can be observed and interpreted. This paper discusses the imagery acquisition task that motivated Hotmap, and presents several examples of information that Hotmap makes visible. we discuss the design choices behind Hotmap, including logarithmic color schemes; low-saturation background images; and tuning images to explore both infrequently-viewed and frequently-viewed spaces.


Examining Associations of Environmental Characteristics with Recreational Cycling Behaviour by Street-Level Strava Data.
Policymakers pay much attention to effectively increasing frequency of people's cycling in the context of developing sustainable and green cities. Investigating associations of environmental characteristics and cycling behaviour could offer implications for changing urban infrastructure aiming at encouraging active travel. However, earlier examinations of associations between environmental characteristics and active travel behaviour are limited by low spatial granularity and coverage of traditional data. Crowdsourced geographic information offers an opportunity to determine the fine-grained travel patterns of people. Particularly, Strava Metro data offer a good opportunity for studies of recreational cycling behaviour as they can offer hourly, daily or annual cycling volumes with different purposes (commuting or recreational) in each street across a city. Therefore, in this study, we utilised Strava Metro data for investigating associations between environmental characteristics and recreational cycling behaviour at a large spatial scale (street level). In this study, we took account of population density, employment density, road length, road connectivity, proximity to public transit services, land use mix, proximity to green space, volume of motor vehicles and traffic accidents in an empirical investigation over Glasgow. Empirical results reveal that Strava cyclists are more likely to cycle for recreation on streets with short length, large connectivity or low volume of motor vehicles or on streets surrounded by residential land.


The use of genetically-engineered animals in science: perspectives of Canadian Animal Care Committee members.
The genetic engineering of animals for their use in science challenges the implementation of refinement and reduction in several areas, including the invasiveness of the procedures involved, unanticipated welfare concerns, and the numbers of animals required. Additionally, the creation of genetically-engineered animals raises problems with the Canadian system of reporting animal numbers per Category of Invasiveness, as well as raising issues of whether ethical limits can, or should, be placed on genetic engineering. A workshop was held with the aim of bringing together Canadian animal care committee members to discuss these issues, to reflect on progress that has been made in addressing them, and to propose ways of overcoming any challenges. Although previous literature has made recommendations with regard to refinement and reduction when creating new genetically-engineered animals, the perception of the workshop participants was that some key opportunities are being missed. The participants identified the main roadblocks to the implementation of refinement and reduction alternatives as confidentiality, cost and competition. If the scientific community is to make progress concerning the implementation of refinement and reduction, particularly in the creation and use of genetically-engineered animals, addressing these roadblocks needs to be a priority.


Methylation of the DPYD promoter: an alternative mechanism for dihydropyrimidine dehydrogenase deficiency in cancer patients.
Dihydropyrimidine dehydrogenase (DPD) deficiency, a known pharmacogenetic syndrome associated with 5-fluorouracil (5-FU) toxicity, has been detected in 3% to 5% of the population. Genotypic studies have identified >32 sequence variants in the DPYD gene; however, in a number of cases, sequence variants could not explain the molecular basis of DPD deficiency. Recent studies in cell lines indicate that hypermethylation of the DPYD promoter might down-regulate DPD expression. The current study investigates the role of methylation in cancer patients with an unexplained molecular basis of DPD deficiency. DPD deficiency was identified phenotypically by both enzyme assay and uracil breath test, and genotypically by denaturing high-performance liquid chromatography. The methylation status was evaluated in PCR products (209 bp) of bisulfite-modified DPYD promoter, using a novel denaturing high-performance liquid chromatography method that distinguishes between methylated and unmethylated alleles. Clinical samples included five volunteers with normal DPD enzyme activity, five DPD-deficient volunteers, and five DPD-deficient cancer patients with a history of 5-FU toxicity. No evidence of methylation was detected in samples from volunteers with normal DPD. Methylation was detected in five of five DPD-deficient volunteers and in three of five of the DPD-deficient cancer patient samples. Of note, one of the two samples from patients with DPD-deficient cancer with no evidence of methylation had the mutation DPYD*2A, whereas the other had DPYD*13. Methylation of the DPYD promoter region is associated with down-regulation of DPD activity in clinical samples and should be considered as a potentially important regulatory mechanism of DPD activity and basis for 5-FU toxicity in cancer patients.


Increase of meningococcal serogroup Y cases in Europe: a reason for concern?
Neisseria meningitidis is differentiated into 12 distinct serogroups of which A, B, C, W-135, X and Y are medically most important and represent a major health problem in one or more parts of the world. The epidemiology of N. meningitidis is unpredictable over time and across geographic regions. A sudden increase or decrease of IMD or meningococcal carriage may occur anywhere at any time. Recent epidemiological surveillance indicates an increase of serogroup Y IMD in some parts of Europe and data from various European countries are presented.


gamma/delta T cell-deficient mice have impaired mucosal immunoglobulin A responses.
Mucosal tissues of mice are enriched in T cells that express the gamma/delta T cell receptor. Since the function of these cells remains unclear, we have compared mucosal immune responses in gamma/delta T cell receptor-deficient (TCRdelta-/-) mice versus control mice of the same genetic background. The frequency of intestinal immunoglobulin (Ig) A plasma cells as well as IgA levels in serum, bile, saliva, and fecal samples were markedly reduced in TCRdelta-/- mice. The TCRdelta-/- mice produced much lower levels of IgA antibodies when immunized orally with a vaccine of tetanus toxoid plus cholera toxin as adjuvant. Conversely, the antigen-specific IgM and IgG antibody responses were comparable to orally immunized control mice. Direct assessment of the cells forming antibodies against the tetanus toxoid and cholera toxin antigens indicated that significantly lower numbers of IgA antibody-producing cells were present in the intestinal lamina propria and Peyer's patches of TCRdelta-/- mice compared with the orally immunized control mice. The selective reduction of IgA responses to ingested antigens in the absence of gamma/delta T cells suggests a specialized role for gamma/delta cells in mucosal immunity.


Genetic Risk for Inflammatory Bowel Disease Is a Determinant of Crohn's Disease Development in Chronic Granulomatous Disease.
Approximately, one-third to one-half of children with chronic granulomatous disease (CGD) develop gastrointestinal inflammation characteristic of idiopathic inflammatory bowel disease (IBD), usually Crohn's disease. We hypothesized that the overall IBD genetic risk, determined by IBD genetic risk score (GRS), might in part determine IBD development in CGD. We reviewed medical records to establish IBD diagnoses in CGD subjects seen at NIAID. IBD risk single nucleotide polymorphism genotypes were determined using the Immunochip, and GRS were estimated by Mangrove. Among 157 white patients with CGD, 55 were confirmed, 78 excluded, and 24 were uncertain for IBD. Two hundred one established, independent European IBD risk single nucleotide polymorphisms passed quality control. After sample quality control and removing non-IBD CGD patients with perianal disease, mean GRS for 40 unrelated patients with CGD-IBD was higher than 53 CGD non-IBD patients (in log2-scale 0.08 ± 1.62 versus -0.67 ± 1.64, P = 0.026) but lower than 239 IBD Genetics Consortium (IBDGC) young-onset Crohn's disease cases (0.76 ± 1.60, P = 0.025). GRS for non-IBD CGD was similar to 609 IBDGC controls (-0.69 ± 1.60, P = 0.95). Seven established IBD single nucleotide polymorphisms were nominally significant among CGD-IBD versus CGD non-IBD, including those near LACC1 (P = 0.005), CXCL14 (P = 0.007), and TNFSF15 (P = 0.016). The weight of the common IBD risk alleles are significant determinants of IBD in CGD. However, IBD risk gene burden among CGD children with IBD is significantly lower than that in nonsyndromic pediatric Crohn's disease, congruent with the concept that defective superoxide production in CGD is also a major IBD risk factor. Individual IBD genes might interact with the CGD defect to cause IBD in CGD.


MiRNA-323-5p Promotes U373 Cell Apoptosis by Reducing IGF-1R.
BACKGROUND MicroRNA regulates mammalian cell growth in terms of its proliferation and apoptosis by controlling the expression of target genes. MiRNA-323-5p plays an important role in regulating cell growth and death within various types of cells. The function of miRNA-323-5p and its possible molecular mechanism in human cerebral glioma U373 cells remains to be further confirmed. The aim of this study was to investigate the regulation function of miRNA-323-5p in human glioma U373 cell growth, proliferation, and apoptosis. MATERIAL AND METHODS We used human cerebral glioma U373 cells as the cell model; utilized liposome technology (transfected by Lipofectamine2000) in human cerebral glioma U373 cells to over-express miRNA-323-5p (microRNA used as control group); and selected MTT assay and flow cytometry to detect cell growth, proliferation, and apoptosis. We used RT-PCR and Western blotting techniques to study the expression levels of target insulin-like growth factor 1 (IGF-1) receptor protein in U373 cells transfected with miRNA-323-5p. We used liposome transfection techniques in human cerebral glioma U373 cells to over-express or processed knockdown of IGF-1R by siRNA, and then transferred with miRNA-323-5p, thereby investigating the treated human cerebral glioma U373 cells apoptosis situations. RESULTS The over-expression of miRNA-323-5p inhibited the growth and proliferation of human cerebral glioma U373 cells and promoted its apoptosis. The over-expression of miRNA-323-5p also reduced the IGF-1R level. After processing the knockdown of IGF-1R and then transfection with miRNA-323-5p, U373 cells had enhanced apoptosis. The over-expression of IGF-1R inhibited the cells apoptosis induced by miRNA-323-5p. CONCLUSIONS MiRNA-323-5p inhibited human cerebral glioma U373 cell proliferation and promoted its apoptosis by reducing IGF-1R.


"Knee osteoarthritis and current research for evidence-are we on the right way?"
An estimated 85% of research is of limited value or wasted because the wrong research questions are addressed. We sought to identify research gaps using American Academy of Orthopaedic Surgeon (AAOS) clinical practice guidelines Treatment of Osteoarthritis of the Knee and Surgical Management of Osteoarthritis of the Knee. Using these recommendations, we conducted searches of ClinicalTrials.gov to discover the extent to which new and ongoing research addresses areas of deficiency. For each recommendation in the AAOS guidelines, we created participants, intervention, comparator, outcomes questions, and search strings using a systematic process. Searches were then conducted of ClinicalTrials.gov to locate relevant studies. Our searches of ClinicalTrials.gov returned 945 studies for surgical and 1416 for non-surgical management of osteoarthritis. Of the 945 studies returned using our search string for surgical trials, 186 (20%) were relevant to 30 (79%) of the 38 recommendations made within the surgical management guideline. Of the 1416 studies returned using our search for non-surgical trials, 360 (25%) were relevant to 16 (89%) of the 18 recommendations made within the conservative management guideline. The development of clinical practice guidelines is a unique opportunity to simultaneously redefine day-to-day decision-making and provide a critical analysis of the status of the literature. Upon our search of the literature since the guidelines introduction, we have found that some inconclusive areas have received more attention than others. Our results should guide researchers towards conducting research on the topics most in need and, by doing so, strengthen the clinical practice guideline recommendations.


A spatially explicit estimate of the prewhaling abundance of the endangered North Atlantic right whale.
The North Atlantic right whale (NARW) (Eubalaena glacialis) is one of the world's most threatened whales. It came close to extinction after nearly a millennium of exploitation and currently persists as a population of only approximately 500 individuals. Setting appropriate conservation targets for this species requires an understanding of its historical population size, as a baseline for measuring levels of depletion and progress toward recovery. This is made difficult by the scarcity of records over this species' long whaling history. We sought to estimate the preexploitation population size of the North Atlantic right whale and understand how this species was distributed across its range. We used a spatially explicit data set on historical catches of North Pacific right whales (NPRWs) (Eubalaena japonica) to model the relationship between right whale relative density and the environment during the summer feeding season. Assuming the 2 right whale species select similar environments, we projected this model to the North Atlantic to predict how the relative abundance of NARWs varied across their range. We calibrated these relative abundances with estimates of the NPRW total prewhaling population size to obtain high and low estimates for the overall NARW population size prior to exploitation. The model predicted 9,075-21,328 right whales in the North Atlantic. The current NARW population is thus <6% of the historical North Atlantic carrying capacity and has enormous potential for recovery. According to the model, in June-September NARWs concentrated in 2 main feeding areas: east of the Grand Banks of Newfoundland and in the Norwegian Sea. These 2 areas may become important in the future as feeding grounds and may already be used more regularly by this endangered species than is thought.


CDC20 assists its catalytic incorporation in the mitotic checkpoint complex.
Open (O) and closed (C) topologies of HORMA-domain proteins are respectively associated with inactive and active states of fundamental cellular pathways. The HORMA protein O-MAD2 converts to C-MAD2 upon binding CDC20. This is rate limiting for assembly of the mitotic checkpoint complex (MCC), the effector of a checkpoint required for mitotic fidelity. A catalyst assembled at kinetochores accelerates MAD2:CDC20 association through a poorly understood mechanism. Using a reconstituted SAC system, we discovered that CDC20 is an impervious substrate for which access to MAD2 requires simultaneous docking on several sites of the catalytic complex. Our analysis indicates that the checkpoint catalyst is substrate assisted and promotes MCC assembly through spatially and temporally coordinated conformational changes in both MAD2 and CDC20. This may define a paradigm for other HORMA-controlled systems.


Renal response to atrial natriuretic factor in rats without intact papillae.
In order to examine the contribution of renal papilla to atrial natriuretic factor (ANF)-induced diuresis, clearance studies were done in rats with intact papillae and bromoethylamine hydrobromide (BEA)-induced papillary necrosis. Papillary necrosis did not attenuate the diuretic and natriuretic response to ANF. In a moderately hydropenic state, urine flow and fractional water excretion were rather more increased in papillary necrosis rats than in papilla-intact ones. In a volume-expanded state, both groups had a similar diuretic response to ANF. Present data indicate that the existence of intact renal papilla is not essential for ANF-induced diuresis. Furthermore, it is suggested that absolute magnitude of ANF-induced diuresis may be dependent on its extrapapillary action(s).


Multimerin is found in the alpha-granules of resting platelets and is synthesized by a megakaryocytic cell line.
In this report, we describe the intracellular localization of multimerin in platelets and its biosynthesis by Dami cells, a megakaryocytic cell line. Immunoelectron microscopy was used to examine frozen thin sections of resting and activated platelets. Multimerin was localized within the platelet alpha-granule in an eccentric position. Within activated platelets, multimerin was found in the surface-connected open cannalicular system and on the external plasma membrane. Light microscopic immunocytochemistry demonstrated multimerin in normal megakaryocytes and in Dami cells after stimulation with PMA. Confirmation of multimerin biosynthesis by Dami cells was obtained by metabolic labeling studies. Both platelet and Dami cell multimerin demonstrated several subunit sizes on reduced SDS-PAGE. However, peptide mapping confirmed structural homology between the different multimerin subunits. Glycosidase digestion demonstrated that multimerin is heavily glycosylated with mainly complex, N-linked carbohydrate. In contrast to the multimerin isolated from platelets, cultured Dami cells secreted mainly smaller multimers of the protein. Biosynthesis of multimerin by a megakaryocytic cell line supports endogenous biosynthesis by megakaryocytes as the origin of this platelet alpha-granule protein.


Clinical features of spontaneous intracerebral hemorrhage in Hispanics and non-Hispanic Whites in New Mexico: a community study.
To compare clinical features of spontaneous intracerebral hemorrhage (ICH) between Hispanics and non-Hispanic Whites in Albuquerque, New Mexico, occurring during the year 1993. Analysis of data collected during a study of ICH incidence in a community. Review medical records with multiple relevant cerebrovascular diagnostic discharge codes in all 9 acute care hospitals in Albuquerque, and in the records of the State Medical Examiner. All suspected cases were verified by cerebral computed tomography or autopsy. Vascular risk factors, test results, and acute outcome were abstracted. There were 38 Hispanics and 46 non-Hispanic Whites with ICH. We found no statistically significant differences between these two ethnic groups in the prevalence of hypertension or diabetes mellitus, in hematoma volume, in seasonal fluctuation of ICH incidence, or in acute mortality. However, based on our sample size, only large differences in the prevalence of risk factors between these two ethnic groups could be detected with statistical significance (>20-30 percentage points). There was a trend toward a higher proportion of subcortical ICH (centered in the basal ganglia, brainstem, or cerebellum) in Hispanics (82%) than in non-Hispanic Whites (62%, P=.09). The higher proportion of subcortical ICH among the Hispanics suggests that chronic hypertension may play a greater role as a risk factor for ICH in this ethnic group than in non-Hispanic Whites in New Mexico. Our findings should be confirmed by larger community studies.


Specialization and niche overlap across spatial scales: Revealing ecological factors shaping species richness and coexistence in Australian songbirds.
Ecological specialization enables the partitioning of resources and thus can facilitate the coexistence of species and promote higher species richness. Specialization and niche partitioning are expected to exert a decisive influence on local spatial scales, while species richness at regional scales should be shaped mostly by historical factors and abiotic conditions. Moreover, specialization is expected to be particularly important in communities that are exceptionally species rich for their environmental conditions. Concurrently, niche overlap in these communities should be minimized to enable species coexistence. We tested these hypotheses by studying specialization-richness relationship and niche overlap in assemblages of 298 species of songbirds (Passeriformes) across Australia. We used local (2-6 ha) to regional (bioregions) spatial scales and detailed data on habitat, diet and foraging behaviour (method, substrate and stratum). We expected the richness-specialization relationship to be particularly strong (a) on local spatial scales and (b) in communities exceptionally species rich for given environmental conditions (approximated by moisture and vegetation complexity). We also expected (c) low niche overlap in assemblages with specialized species. Only the third prediction was partly supported. First, while the specialization and species richness were often positively related, the strength and the direction of the relationship changed between traits and across spatial scales. The strength of the specialization-richness relationship was consistently positive only in foraging stratum, and it increased towards smaller spatial scales only in case of habitat and diet. Simultaneously, species in local communities demonstrated high overlap in habitat and diet. Second, we did not find particularly strong specialization-richness relationships in exceptionally species-rich communities. Third, we found the expected negative relationship between specialization and overlap in foraging stratum and substrate (in local communities), suggesting that species partition ecological space locally in terms of where they find food. Our expectations were only weakly supported. Specialization on foraging stratum was probably important in facilitating species coexistence. Conversely, although species were often specialized on habitat and diet, high overlap in these traits did not preclude their local coexistence. Overall, specialization and overlap in foraging traits were more important for species coexistence than habitat or diet.


Brain structure. Cell types in the mouse cortex and hippocampus revealed by single-cell RNA-seq.
The mammalian cerebral cortex supports cognitive functions such as sensorimotor integration, memory, and social behaviors. Normal brain function relies on a diverse set of differentiated cell types, including neurons, glia, and vasculature. Here, we have used large-scale single-cell RNA sequencing (RNA-seq) to classify cells in the mouse somatosensory cortex and hippocampal CA1 region. We found 47 molecularly distinct subclasses, comprising all known major cell types in the cortex. We identified numerous marker genes, which allowed alignment with known cell types, morphology, and location. We found a layer I interneuron expressing Pax6 and a distinct postmitotic oligodendrocyte subclass marked by Itpr2. Across the diversity of cortical cell types, transcription factors formed a complex, layered regulatory code, suggesting a mechanism for the maintenance of adult cell type identity.


Morphological and functional characterization of beta TC-6 cells--an insulin-secreting cell line derived from transgenic mice.
Morphological analysis of hormone content and functional assessment of hormone secretion were conducted in beta TC-6 cells, an insulin-secreting cell line derived from transgenic mice expressing the large T-antigen of simian virus 40 (SV40) in pancreatic beta-cells. We observed by immunohistochemistry and confocal microscopy that beta TC-6 cells contain abundant insulin and small amounts of glucagon and somatostatin (SRIF). Glucagon usually co-localized with insulin, whereas cells containing SRIF did not contain insulin or glucagon. Static incubation and perifusion experiments demonstrated that beta TC-6 cells at passage 30-45 secrete insulin in response to glucose. In static incubations, maximal stimulation was achieved for glucose concentrations > 2.8 mmol/l glucose, and the half-maximal effect was observed at 0.5 mmol/l. Maximal stimulation was four times greater than HIT-T15 cells at passage 72-81, although HIT cells had a greater response over their basal levels. The magnitude of the insulin response to glucose in perifusion was 1,734 +/- 384 pmol.l-1. min and was 4.6-fold greater in the presence of 3-isobutyl-1-methylxanthine. Low amounts of glucagon were released in response to amino acids. Epinephrine (EPI), and to a lesser extent SRIF, inhibited phasic glucose-induced insulin secretion. A major portion of these inhibitory effects was mediated by pertussis toxin-sensitive substrates. Immunoblots detected the presence of the G-proteins Gi alpha 2, Gi alpha 3, and Go alpha 2. These results indicate that beta TC-6 cells are a glucose-responsive cell line in which insulin exocytosis is physiologically regulated by EPI and SRIF through Gi/Go-mediated mechanisms.


Evidence-based suggestions for management of air leaks.
The management of postoperative alveolar air leaks (AALs) continues to challenge thoracic surgeons. AALs increase length of stay and health care costs, and likely lead to other postoperative complications. Staple line buttresses, topical sealants, pleural tents, pneumoperitoneum, and modifications of traditional chest tube management (ie, reduced suction) have all been proposed to help reduce AAL. However, the cost of some of the commercial products being marketed may outweigh their relative effectiveness, and some of these techniques and products have not been adequately studied to date. This article provides a review of the available evidence-based literature that addresses the efficacy of the options currently available to prevent and manage AALs. Management suggestions based on this literature are presented.


Structure and Composition of Native Membrane Derived Polymer-Supported Lipid Bilayers.
Over the last two decades, supported lipid bilayers (SLBs) have been extensively used as model systems to study cell membrane structure and function. While SLBs have been traditionally produced from simple lipid mixtures, there has been a recent surge in compositional complexity to better mimic cellular membranes and thereby bridge the gap between classic biophysical approaches and cell experiments. To this end, native cellular membrane derived SLBs (nSLBs) have emerged as a new category of SLBs. As a new type of biomimetic material, an analytical workflow must be designed to characterize its molecular composition and structure. Herein, we demonstrate how a combination of fluorescence microscopy, neutron reflectometry, and secondary ion mass spectrometry offers new insights on structure, composition, and quality of nSLB systems formed using so-called hybrid vesicles, which are a mixture of native membrane material and synthetic lipids. With this approach, we demonstrate that the nSLB formed a continuous structure with complete mixing of the synthetic and native membrane components and a molecular stoichiometry that essentially mirrors that of the hybrid vesicles. Furthermore, structural investigation of the nSLB revealed that PEGylated lipids do not significantly thicken the hydration layer between the bilayer and substrate when on silicon substrates; however, nSLBs do have more topology than their simpler, purely synthetic counterparts. Beyond new insights regarding the structure and composition of nSLB systems, this work also serves to guide future researchers in producing and characterizing nSLBs from their cellular membrane of choice.


Immunohistochemical demonstration of glutathione S-transferases in primary human breast carcinomas.
The expression of cytosolic glutathione S-transferase (GST) isoenzymes has been assessed in a series of 74 primary human breast carcinomas using an immunohistochemical method. GST pi was detected in sections from all 74 tumours; it was expressed by non-epithelial (stromal and inflammatory) cells in 62 tumours (84 per cent), but by tumour epithelium in only 35 (47 per cent). Non-neoplastic mammary epithelium was uniformly positive for GST pi. Expression of GST alpha and mu was observed in 19 and 42 per cent of the tumours, respectively, and was largely confined to the neoplastic component. Lack of staining of tumour epithelium for GST pi was significantly associated with poorer tumour differentiation (higher grade). There was no association between expression of any of the three isoenzymes and either menopausal status or expression of c-erbB-2 oncogene protein product. Immunohistochemistry is a useful method for the investigation of expression and cellular localization of GSTs within tumours; such data are needed to improve our understanding of the role of these enzymes in neoplasia and in resistance to cytotoxic drug therapy.


The relevance of HLA sequencing in population genetics studies.
Next generation sequencing (NGS) is currently being adapted by different biotechnological platforms to the standard typing method for HLA polymorphism, the huge diversity of which makes this initiative particularly challenging. Boosting the molecular characterization of the HLA genes through efficient, rapid, and low-cost technologies is expected to amplify the success of tissue transplantation by enabling us to find donor-recipient matching for rare phenotypes. But the application of NGS technologies to the molecular mapping of the MHC region also anticipates essential changes in population genetic studies. Huge amounts of HLA sequence data will be available in the next years for different populations, with the potential to change our understanding of HLA variation in humans. In this review, we first explain how HLA sequencing allows a better assessment of the HLA diversity in human populations, taking also into account the methodological difficulties it introduces at the statistical level; secondly, we show how analyzing HLA sequence variation may improve our comprehension of population genetic relationships by facilitating the identification of demographic events that marked human evolution; finally, we discuss the interest of both HLA and genome-wide sequencing and genotyping in detecting functionally significant SNPs in the MHC region, the latter having also contributed to the makeup of the HLA molecular diversity observed today.


An Interesting Case of Carbamazepine-Induced Stevens-Johnson Syndrome.
A 29-year-old Black female patient was admitted to a psychiatric ward with symptoms of major depressive disorder with psychosis. The patient was started on amitriptyline 50 mg/day and haloperidol 10 mg/day. On day 4 post-admission, the preferred first-line antidepressant, fluoxetine, became available and the patient was switched from amitriptyline to fluoxetine 20 mg/day. On the same day, the dose of haloperidol was reduced to 5 mg/day. Thirteen days post-initiation of these medications the patient became talkative, associated with emotional lability, an expansive mood, irritability and restlessness. The working diagnosis was changed to bipolar affective disorder in the manic phase. Fluoxetine was discontinued and carbamazepine 600 mg/day was added to the patient's treatment regimen. Her manic symptoms started to resolve; however, 14 days post-initiation of carbamazepine, the patient had a fever; itchy, discharging eyes; respiratory distress; generalised symmetrical erythematosus rash; buccal ulceration; and conjunctival injection with difficulty opening her eyes. Carbamazepine was immediately discontinued and the patient received intravenous fluid resuscitation. The patient recovered considerably after 12 days of symptomatic and supportive management, and was transferred back to the psychiatric ward for the continuation of bipolar disorder management. Lithium therapy was instituted and the patient was subsequently discharged. Using the Algorithm of Drug causality for Epidermal Necrolysis (ALDEN) Stevens-Johnson Syndrome/toxic epidermal necrolysis (SJS/TEN) drug causality scoring system, carbamazepine and fluoxetine were evaluated as 'very probable' and 'possible' causes of SJS, respectively, in this patient. Fluoxetine-induced SJS was considered on account of previous case reports, however no evidence of causality was found in this patient. Consecutive administration with a potential increase in carbamazepine due to inhibition of cytochrome P450 (CYP) 3A4 metabolism by fluoxetine was also not ruled out. A diagnosis of carbamazepine-induced SJS was made and was considered an idiosyncratic adverse drug reaction.


Dismantling myths on the airborne transmission of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2).
The coronavirus disease 2019 (COVID-19) pandemic has caused untold disruption throughout the world. Understanding the mechanisms for transmission of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is key to preventing further spread, but there is confusion over the meaning of 'airborne' whenever transmission is discussed. Scientific ambivalence originates from evidence published many years ago which has generated mythological beliefs that obscure current thinking. This article collates and explores some of the most commonly held dogmas on airborne transmission in order to stimulate revision of the science in the light of current evidence. Six 'myths' are presented, explained and ultimately refuted on the basis of recently published papers and expert opinion from previous work related to similar viruses. There is little doubt that SARS-CoV-2 is transmitted via a range of airborne particle sizes subject to all the usual ventilation parameters and human behaviour. Experts from specialties encompassing aerosol studies, ventilation, engineering, physics, virology and clinical medicine have joined together to produce this review to consolidate the evidence for airborne transmission mechanisms, and offer justification for modern strategies for prevention and control of COVID-19 in health care and the community.


Healthcare workers' perceptions of the duty to work during an influenza pandemic.
Healthcare workers (HCWs) are often assumed to have a duty to work, even if faced with personal risk. This is particularly so for professionals (doctors and nurses). However, the health service also depends on non-professionals, such as porters, cooks and cleaners. The duty to work is currently under scrutiny because of the ongoing challenge of responding to pandemic influenza, where an effective response depends on most uninfected HCWs continuing to work, despite personal risk. This paper reports findings of a survey of HCWs (n = 1032) conducted across three National Health Service trusts in the West Midlands, UK, to establish whether HCWs' likelihood of working during a pandemic is associated with views about the duty to work. The sense that HCWs felt that they had a duty to work despite personal risk emerged strongly regardless of professional status. Besides a strong sense that everyone should pull together, all kinds of HCWs recognised a duty to work even in difficult circumstances, which correlated strongly with their stated likelihood of working. This suggests that HCWs' decisions about whether or not they are prepared to work during a pandemic are closely linked to their sense of duty. However, respondents' sense of the duty to work may conflict with their sense of duty to family, as well as other factors such as a perceived lack of reciprocity from their employers. Interestingly, nearly 25% of doctors did not consider that they had a duty to work where doing so would pose risks to themselves or their families.


Sperm counts and sperm sex ratio in male infertility patients.
In recent years, investigators have noted a trend toward a declining proportion of male births in many industrialized nations. While men bear the sex-determining chromosome, the role of the female partner as it pertains to fertilization or miscarriage may also alter the gender ratio. We attempted to determine a man's secondary sex ratio (F1 generation) by directly examining the sex chromosomes of his sperm. We examined our male infertility clinic database for all men who had undergone a semen fluorescence in situ hybridization (FISH). Patient demographic and semen parameters were recorded. Chi-squared analysis was used to compare gender ratios (Y chromosomes/total chromosomes). Multivariable logistic regression was used to predict the odds of possessing a Y-bearing sperm after accounting for demographic and semen parameters. A total of 185 men underwent sperm FISH. For the entire cohort, the proportion of Y chromosome-bearing sperm was 51.5%. Men with less than five million motile sperm had a significantly lower proportion of Y chromosome-bearing sperm (50.8%) compared to men with higher sperm counts (51.6%; P=0.02). After multivariable adjustment, a higher sperm concentration, total motile sperm count and semen volume significantly increased the odds of having a Y chromosome-bearing sperm (P<0.01). As a man's sperm production declines, so does the proportion of Y chromosome-bearing sperm. Thus, a man's reproductive potential may predict his ability to sire male offspring.


Health literacy assessment using talking touchscreen technology (Health LiTT): a new item response theory-based measure of health literacy.
The importance of health literacy has grown considerably among researchers, clinicians, patients, and policymakers. Better instruments and measurement strategies are needed. Our objective was to develop a new health literacy instrument using novel health information technology and modern psychometrics. We designed Health LiTT as a self-administered multimedia touchscreen test based on item response theory (IRT) principles. We enrolled a diverse group of 619 English-speaking, primary care patients in clinics for underserved patients. We tested three item types (prose, document, quantitative) that worked well together to reliably measure a single dimension of health literacy. The Health LiTT score meets psychometric standards (reliability of 0.90 or higher) for measurement of individual respondents in the low to middle range. Mean Health LiTT scores were associated with age, race/ethnicity, education, income, and prior computer use (p < .05). We created an IRT-calibrated item bank of 82 items. Standard setting needs to be performed to classify and map items onto the construct and to identify measurement gaps. We are incorporating Health LiTT into an existing online research management tool. This will enable administration of Health LiTT on the same touchscreen used for other patient-reported outcomes, as well as real-time scoring and reporting of health literacy scores.


Effect of a postnatal high-fat diet exposure on puberty onset, estrous cycle regularity, and kisspeptin expression in female rats.
Kisspeptin, encoded by Kiss1, plays a key role in pubertal maturation and reproduction as a positive upstream regulator of the hypothalamic-pituitary-gonadal (HPG) axis. To examine the role of high-fat diet (HFD) on puberty onset, estrous cycle regularity, and kisspeptin expression, female rats were exposed to HFD in distinct postnatal periods. Three groups of rats were exposed to HFD containing 60% energy from fat during the pre-weaning period (postnatal day (PND) 1-16, HFD PND 1-16), post-weaning period (HFD PND 21-34), or during both periods (HFD PND 1-34). Puberty onset, evaluated by vaginal opening, was monitored on days 30-34. Leptin, estradiol (E2), Kiss1 mRNA levels, and number of kisspeptin-immunoreactive cells in the anteroventral periventricular nucleus (AVPV) and arcuate nucleus (ARC) were measured at day 34. Body weight increased only in rats exposed to HFD during post-weaning period, whereas the timing of vaginal opening was unaffected in all three groups. Leptin, Kiss1 mRNA levels, and number of kisspeptin-immunoreactive cells at day 34 were not affected by HFD. Additionally, the estrous cycle regularity was monitored in rats exposed to HFD for 40 days from weaning. Leptin, E2, and Kiss1 mRNA levels in the AVPV and ARC were measured after the HFD exposure. Thirty-three percent of rats exposed to HFD exhibited irregular estrous cycles and a two-fold increase in leptin. By contrast, E2 level and Kiss1 mRNA levels were not affected by the treatment. These data show that postnatal HFD exposure induced irregular estrous cycles, but had no effect on puberty onset or kisspeptin.


Active holography in InGaAs/InP quantum-well microcavities.
High-efficiency dynamic holography at 1.55 μm is demonstrated in a broad-area InGaAs/InP multiple-quantum-well vertical microcavity. The design places single quantum wells at the cavity antinodes, reducing mode-pulling and enabling a higher Q-factor. The device is pumped by interference fringes through an amorphous mirror that is transparent to a high-energy hologram writing pulse at a wavelength of 1.06 μm. Optically pumped free carrier gratings are probed by a tunable 1.5 μm laser in a four-wave mixing configuration. Diffraction efficiency into both m=±1 diffraction orders of 35% (70% total) has been obtained with a phase grating contribution approaching the maximum π phase shift by combining absorption bleaching with asymmetric Fabry-Perot reflectivity. The diffracted signal exhibits rise/fall times of 5 ns, demonstrating the high speed capabilities of this device.


Area-level deprivation and overall and cause-specific mortality: 12 years' observation on British women and systematic review of prospective studies.
Prospective studies have suggested a negative impact of area deprivation on overall mortality, but its effect on cause-specific mortality and the mechanisms that account for this association remain unclear. We investigate the association of area deprivation, using Index of Multiple deprivation (IMD), with overall and cause-specific mortality, contextualising findings within a systematic review. We used data from 4,286 women from the British Women's Heart Health Study (BWHHS) recruited at 1999-2001 to examine the association of IMD with overall and cause-specific mortality using Cox regression models. One standard deviation (SD) increase in the IMD score had a hazard ratio (HR) of 1.21 (95% CI: 1.13-1.30) for overall mortality after adjustment for age and lifecourse individual deprivation, which was attenuated to 1.15 (95% CI: 1.04-1.26) after further inclusion of mediators (health behaviours, biological factors and use of statins and blood pressure-lowering medications). A more pronounced association was observed for respiratory disease and vascular deaths. The meta-analysis, based on 20 published studies plus the BWHHS (n=21), yielded a summary relative risk (RR) of 1.15 (95% CI: 1.11-1.19) for area deprivation (top [least deprived; reference] vs. bottom tertile) with overall mortality in an age and sex adjusted model, which reduced to 1.06 (95% CI: 1.04-1.08) in a fully adjusted model. Health behaviours mediate the association between area deprivation and cause-specific mortality. Efforts to modify health behaviours may be more successful if they are combined with measures that tackle area deprivation.


Surgery--still an "old boys' club"?
Currently more than 40% of first-year medical students are female, yet less than 20% of those entering surgical residencies are women. It has been suggested that the surgical environment experienced during medical school clinical clerkships may be perceived as being unfavorable to female students, thus accounting for this disparity. One hundred and five medical students at The Johns Hopkins University School of Medicine responded to a questionnaire administered before graduation to assess career choice, the influence of a number of conditions on choice of specialty, the perception of a specialty's attitudes toward their gender, and the students' experiences during clinical rotations. Thirty-four percent of men and 13% of women chose surgery or one of its subspecialties as their career (p < 0.01). Eighty-seven percent of women responding perceived a specialty as unfavorable toward their gender versus 21% of men (p < 0.0001). None of the men believed that surgery was unfavorable toward their gender, whereas 96% of these women believed that surgery was unfavorable (p < 0.00001). Fifty percent of women felt out of place on a clinical service, with 92% of these having this perception on a surgical service. Only 9% of men ever felt out of place on a clinical service (p < 0.0001), and none of the men felt out of place in surgery (p < 0.0001). Female medical students perceive a gender bias on surgical services, suggesting that an "old boys' club" attitude may still exist in surgery.


Pertussis toxin, an inhibitor of G(αi) PCR, inhibits bile acid- and cytokine-induced apoptosis in primary rat hepatocytes.
Excessive hepatocyte apoptosis is a common event in acute and chronic liver diseases leading to loss of functional liver tissue. Approaches to prevent apoptosis have therefore high potential for the treatment of liver disease. G-protein coupled receptors (GPCR) play crucial roles in cell fate (proliferation, cell death) and act through heterotrimeric G-proteins. G(αi)PCRs have been shown to regulate lipoapoptosis in hepatocytes, but their role in inflammation- or bile acid-induced apoptosis is unknown. Here, we analyzed the effect of inhibiting G(αi)PCR function, using pertussis toxin (PT), on bile acid- and cytokine-induced apoptosis in hepatocytes. Primary rat hepatocytes, HepG2-rNtcp cells (human hepatocellular carcinoma cells) or H-4-II-E cells (rat hepatoma cells) were exposed to glycochenodeoxycholic acid (GCDCA) or tumor necrosis factor-α (TNFα)/actinomycin D (ActD). PT (50-200 nmol/L) was added 30 minutes prior to the apoptotic stimulus. Apoptosis (caspase-3 activity, acridine orange staining) and necrosis (sytox green staining) were assessed. PT significantly reduced GCDCA- and TNFα/ActD-induced apoptosis in rat hepatocytes (-60%, p<0.05) in a dose-dependent manner (with no shift to necrosis), but not in HepG2-rNtcp cells or rat H-4-II-E cells. The protective effect of pertussis toxin was independent of the activation of selected cell survival signal transduction pathways, including ERK, p38 MAPK, PI3K and PKC pathways, as specific protein kinase inhibitors did not reverse the protective effects of pertussis toxin in GCDCA-exposed hepatocytes. Pertussis toxin, an inhibitor of G(αi)PCRs, protects hepatocytes, but not hepatocellular carcinoma cells, against bile acid- and cytokine-induced apoptosis and has therapeutic potential as primary hepatoprotective drug, as well as adjuvant in anti-cancer therapy.


A composite polymer nanoparticle overcomes multidrug resistance and ameliorates doxorubicin-associated cardiomyopathy.
Acquired chemotherapy resistance is a major contributor to treatment failure in oncology. For example, the efficacy of the common anticancer agent doxorubicin (DOX) is limited by the emergence of multidrug resistance (MDR) phenotype in cancer cells. While dose escalation of DOX can circumvent such resistance to a degree, this is precluded by the appearance of cardiotoxicity, a particularly debilitating condition in children. In vitro studies have established the ability of the natural phytochemical curcumin to overcome MDR; however, its widespread clinical application is restricted by poor solubility and low bioavailability. Building upon our recently developed polymer nanoparticle of curcumin (NanoCurc or NC) that significantly enhances the systemic bioavailability of curcumin, we synthesized a doxorubicin-curcumin composite nanoparticle formulation called NanoDoxCurc (NDC) for overcoming DOX resistance. Compared to DOX alone, NDC inhibited the MDR phenotype and caused striking growth inhibition both in vitro and in vivo in several models of DOX-resistant cancers (multiple myeloma, acute leukemia, prostate and ovarian cancers, respectively). Notably, NDC-treated mice also demonstrated complete absence of cardiac toxicity, as assessed by echocardiography, or any bone marrow suppression, even at cumulative dosages where free DOX and pegylated liposomal DOX (Doxil®) resulted in demonstrable attenuation of cardiac function and hematological toxicities. This improvement in safety profile was achieved through a reduction of DOX-induced intracellular oxidative stress, as indicated by total glutathione levels and glutathione peroxidase activity in cardiac tissue. A composite DOX-curcumin nanoparticle that overcomes both MDR-based DOX chemoresistance and DOX-induced cardiotoxicity holds promise for providing lasting and safe anticancer therapy.


Structural model of the dimeric Parkinson's protein LRRK2 reveals a compact architecture involving distant interdomain contacts.
Leucine-rich repeat kinase 2 (LRRK2) is a large, multidomain protein containing two catalytic domains: a Ras of complex proteins (Roc) G-domain and a kinase domain. Mutations associated with familial and sporadic Parkinson's disease (PD) have been identified in both catalytic domains, as well as in several of its multiple putative regulatory domains. Several of these mutations have been linked to increased kinase activity. Despite the role of LRRK2 in the pathogenesis of PD, little is known about its overall architecture and how PD-linked mutations alter its function and enzymatic activities. Here, we have modeled the 3D structure of dimeric, full-length LRRK2 by combining domain-based homology models with multiple experimental constraints provided by chemical cross-linking combined with mass spectrometry, negative-stain EM, and small-angle X-ray scattering. Our model reveals dimeric LRRK2 has a compact overall architecture with a tight, multidomain organization. Close contacts between the N-terminal ankyrin and C-terminal WD40 domains, and their proximity-together with the LRR domain-to the kinase domain suggest an intramolecular mechanism for LRRK2 kinase activity regulation. Overall, our studies provide, to our knowledge, the first structural framework for understanding the role of the different domains of full-length LRRK2 in the pathogenesis of PD.


Verification and unmasking of widely used human esophageal adenocarcinoma cell lines.
For decades, hundreds of different human tumor type-specific cell lines have been used in experimental cancer research as models for their respective tumors. The veracity of experimental results for a specific tumor type relies on the correct derivation of the cell line. In a worldwide effort, we verified the authenticity of all available esophageal adenocarcinoma (EAC) cell lines. We proved that the frequently used cell lines SEG-1 and BIC-1 and the SK-GT-5 cell line are in fact cell lines from other tumor types. Experimental results based on these contaminated cell lines have led to ongoing clinical trials recruiting EAC patients, to more than 100 scientific publications, and to at least three National Institutes of Health cancer research grants and 11 US patents, which emphasizes the importance of our findings. Widespread use of contaminated cell lines threatens the development of treatment strategies for EAC.


Use of an oxonol dye in combination with confocal laser scanning microscopy to monitor damage to Staphylococcus aureus cells during colonisation of silver-coated vascular grafts.
The antimicrobial silver-coating of medical prostheses is regarded as a means to reduce the risk of bacterial colonisation after implantation. The effect of a silver-coating of vascular grafts on biofilm formation was assessed in batch cultures of Staphylococcus aureus, using confocal laser scanning microscopy. Total cells in biofilms were analysed by staining with the DNA-binding fluorochrome SYTO 62 and the proportion of damaged cells was quantified with the membrane potential-sensitive dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol. Both the extent of biofilm formation and the proportion of viable biofilm cells were significantly diminished on the surface of the silver-coated vascular grafts compared with uncoated controls, probably due to the antimicrobial activity of silver ions released from the silver-coated graft surface.


Timing as a sexually selected trait: the right mate at the right moment.
Sexual selection favours the expression of traits in one sex that attract members of the opposite sex for mating. The nature of sexually selected traits such as vocalization, colour and ornamentation, their fitness benefits as well as their costs have received ample attention in field and laboratory studies. However, sexually selected traits may not always be expressed: coloration and ornaments often follow a seasonal pattern and behaviours may be displayed only at specific times of the day. Despite the widely recognized differences in the daily and seasonal timing of traits and their consequences for reproductive success, the actions of sexual selection on the temporal organization of traits has received only scant attention. Drawing on selected examples from bird and mammal studies, here we summarize the current evidence for the daily and seasonal timing of traits. We highlight that molecular advances in chronobiology have opened exciting new opportunities for identifying the genetic targets that sexual selection may act on to shape the timing of trait expression. Furthermore, known genetic links between daily and seasonal timing mechanisms lead to the hypothesis that selection on one timescale may simultaneously also affect the other. We emphasize that studies on the timing of sexual displays of both males and females from wild populations will be invaluable for understanding the nature of sexual selection and its potential to act on differences within and between the sexes in timing. Molecular approaches will be important for pinpointing genetic components of biological rhythms that are targeted by sexual selection, and to clarify whether these represent core or peripheral components of endogenous clocks. Finally, we call for a renewed integration of the fields of evolution, behavioural ecology and chronobiology to tackle the exciting question of how sexual selection contributes to the evolution of biological clocks.This article is part of the themed issue 'Wild clocks: integrating chronobiology and ecology to understand timekeeping in free-living animals'.


Effect of extract of Rhazya stricta, a traditional medicinal plant, on rat brain tribulin.
Rhazya stricta leaves, which have both antidepressant and sedative properties in animal models, are widely used in folk medicine in the Arabian peninsula. In this study, the effects of oral administration of leaf extracts on rat brain tribulin levels [endogenous monoamine oxidase (MAO) A and B inhibitory activity], were determined. In an acute study, low doses brought about an increase in MAO A inhibitory activity, while intermediate doses caused a significant reduction. The highest doses had no significant effects on activity. There were no significant effects on MAO B inhibitory activity at any dose. Subchronic administration (21 days) caused a significant decrease in MAO A inhibitory activity, most prominent at low dosage, and an increase in MAO B inhibitory activity. Acute intramuscular administration also resulted in a similar pattern. Such paradoxical effects were at least partially explained when different extracts of the leaves were used; a weakly basic chloroform fraction caused an increase in MAO A inhibitory activity, whereas butanol extracts brought about a decrease. These fractions had no significant effects on MAO B inhibitory activity. The findings show that Rhazya stricta leaves contain at least two different components that affect MAO inhibitory activity in opposite directions. It may be that the antidepressant and sedative actions of the plant are explicable in terms of these different components.


Archaeal (per)chlorate reduction at high temperature: an interplay of biotic and abiotic reactions.
Perchlorate and chlorate anions [(per)chlorate] exist in the environment from natural and anthropogenic sources, where they can serve as electron acceptors for bacteria. We performed growth experiments combined with genomic and proteomic analyses of the hyperthermophile Archaeoglobus fulgidus that show (per)chlorate reduction also extends into the archaeal domain of life. The (per)chlorate reduction pathway in A. fulgidus relies on molybdo-enzymes that have similarity with bacterial enzymes; however, chlorite is not enzymatically split into chloride and oxygen. Evidence suggests that it is eliminated by an interplay of abiotic and biotic redox reactions involving sulfur compounds. Biological (per)chlorate reduction by ancient archaea at high temperature may have prevented accumulation of perchlorate in early terrestrial environments and consequently given rise to oxidizing conditions on Earth before the rise of oxygenic photosynthesis.


Emotion regulation in psychopathy.
Emotion processing is known to be impaired in psychopathy, but less is known about the cognitive mechanisms that drive this. Our study examined experiencing and suppression of emotion processing in psychopathy. Participants, violent offenders with varying levels of psychopathy, viewed positive and negative images under conditions of passive viewing, experiencing and suppressing. Higher scoring psychopathics were more cardiovascularly responsive when processing negative information than positive, possibly reflecting an anomalously rewarding aspect of processing normally unpleasant material. When required to experience emotional response, by 'getting into the feeling' of the emotion conveyed by a negative image, higher factor 1 psychopathic individuals showed reduced responsiveness, suggesting that they were less able to do this. These data, together with the absence of corresponding differences in subjective self-report might be used to inform clinical strategies for normalising emotion processing in psychopathic offenders to improve treatment outcome, and reduce risk amongst this client group.


Efficacy of rituximab in limited Wegener's granulomatosis with refractory granulomatous manifestations.
Patients with limited Wegener's granulomatosis (WG) may experience a relapsing and remitting course. How such patients should be treated, particularly when they are refractory to standard of care therapies, is not clear. Rituximab is a monoclonal anti-CD20 antibody that has been used successfully to treat multiple forms of autoimmune and rheumatic diseases, but its role in the treatment of limited WG remains uncertain. Eight patients with limited WG who were refractory to (or intolerant of) standard immunosuppressive therapies were evaluated at the Johns Hopkins Hospital or the Mayo Clinic Rochester, and were treated with rituximab using a standard lymphoma protocol. Four men and 4 women with limited WG were treated with rituximab. Patients' mean age was 39 years. All patients had predominantly necrotizing granulomatous disease manifestations, including chronic sinusitis, pulmonary nodules, orbital pseudotumor, and subglottic stenosis. Patients had failed an average of 3 immunosuppressive agents, not including glucocorticoids. Six patients had failed (or were intolerant of) therapy with cyclophosphamide; all 8 had failed therapy with methotrexate. At the time of treatment, 3 of the 8 patients were antineutrophil cytoplasmic antibody-negative. Rituximab successfully induced disease remission in all 8 patients. Three patients were retreated preemptively with rituximab after return of peripheral blood B-cells. Five patients were successfully retreated with rituximab after disease flare. Rituximab is an effective therapy for patients with limited WG and may be sufficient to induce sustained remission, even among patients with refractory disease and predominantly necrotizing granulomatous disease manifestations.