The protective effects of prolactin on brain injury.
Brain injuries based on their causes are divided into two categories, TBI and NTBI. TBI is caused by damages such as head injury, but non-physical injury causes NTBI. Prolactin is one of the blood factors that increase during brain injury. It has been assumed to play a regenerative role in post-injury recovery. In this review, various valid papers from electronic sources (including Web of Science, Scopus, PubMed, SID, Google Scholar, and ISI databases) used, which in them the protective effect of prolactin on brain injury investigated. Inflammation following brain injury with the production of pro-inflammatory cytokines in the affected area can even lead to excitotoxicity and cell death in the damaged area. Medical brain damage treatments are long-term, and can have several side effects. Therefore, it is better to consider medication treatments that have fewer side effects and greater efficacy. Research suggests that prolactin has numerous regenerative effects on brain injury, and prevents cell death. Prolactin is one of the hormones produced in the body; therefore it has fewer side effects and may be more effective because it increases during brain injury. Prolactin can be used peripherally and centrally, and exerts its neuro regenerative effects against further damage post-TBI and NTBI.


B-cell repertoire dynamics after sequential hepatitis B vaccination and evidence for cross-reactive B-cell activation.
A diverse B-cell repertoire is essential for recognition and response to infectious and vaccine antigens. High-throughput sequencing of B-cell receptor (BCR) genes can now be used to study the B-cell repertoire at great depth and may shed more light on B-cell responses than conventional immunological methods. Here, we use high-throughput BCR sequencing to provide novel insight into B-cell dynamics following a primary course of hepatitis B vaccination. Nine vaccine-naïve participants were administered three doses of hepatitis B vaccine (months 0, 1, and 2 or 7). High-throughput Illumina sequencing of the total BCR repertoire was combined with targeted sequencing of sorted vaccine antigen-enriched B cells to analyze the longitudinal response of both the total and vaccine-specific repertoire after each vaccine. ELISpot was used to determine vaccine-specific cell numbers following each vaccine. Deconvoluting the vaccine-specific from total BCR repertoire showed that vaccine-specific sequence clusters comprised <0.1 % of total sequence clusters, and had certain stereotypic features. The vaccine-specific BCR sequence clusters were expanded after each of the three vaccine doses, despite no vaccine-specific B cells being detected by ELISpot after the first vaccine dose. These vaccine-specific BCR clusters detected after the first vaccine dose had distinct properties compared to those detected after subsequent doses; they were more mutated, present at low frequency even prior to vaccination, and appeared to be derived from more mature B cells. These results demonstrate the high-sensitivity of our vaccine-specific BCR analysis approach and suggest an alternative view of the B-cell response to novel antigens. In the response to the first vaccine dose, many vaccine-specific BCR clusters appeared to largely derive from previously activated cross-reactive B cells that have low affinity for the vaccine antigen, and subsequent doses were required to yield higher affinity B cells.


Monensin toxicity in cattle.
Monensin has been tested to determine its toxicity and safety in cattle. Single dose acute toxicity and signs associated with toxicity were determined by oral gavage, 7-d oral gavage and feeding experiments with high concentrations of monensin in feed. Oral feeding studies indicated a near complete anorexia resulting from intake of sublethal amounts of monensin. In these cases, cattle recovered from the insulting dose and resumed growth and feed intake. In long-term chronic feedlot, pasture supplement, and reproduction safety studies conducted with monensin administered in the feed, the high concentrations caused cattle to show signs of mild monensin intoxication. Mortality resulted from feeding groups of cattle large quantities of monensin in small quantities of feed. Furthermore, these studies have demonstrated no detrimental effects upon reproduction. Collectively, these studies indicate that the greatest risk of intoxication occurs when cattle first receive a feed containing monensin. Mixing errors and misuse situations under actual use conditions have resulted in cases of cattle mortality. In most cases the mortality was predictable based upon the exposure in controlled studies.


Impact of acute exposure to WTC dust on ciliated and goblet cells in lungs of rats.
Clinical studies and the World Trade Center (WTC) Health Registry have revealed increases in the incidence of chronic (non-cancer) lung disorders among first responders (FR) who were at Ground Zero during the initial 72 h after the collapse. Our previous analyses of rats exposed to building-derived WTC dusts using exposure scenarios/levels that mimicked FR mouth-breathing showed that a single WTC dust exposure led to changes in expression of genes whose products could be involved in the lung ailments, but few other significant pathologies. We concluded that rather than acting as direct inducers of many of the FR health effects, it was more likely inhaled WTC dusts instead may have impacted on toxicities induced by other rescue-related co-pollutants present in Ground Zero air. To allow for such effects to occur, we hypothesized that the alkaline WTC dusts induced damage to the normal ability of the lungs to clear inhaled particles. To validate this, rats were exposed on two consecutive days (2 h/d, by intratracheal inhalation) to WTC dust (collected 12-13 September 2001) and examined over a 1-yr period thereafter for changes in the presence of ciliated cells in the airways and hyperplastic goblet cells in the lungs. WTC dust levels in the lungs were assessed in parallel to verify that any changes in levels of these cells corresponded with decreases in host ability to clear the particles themselves. Image analyses of the rat lungs revealed a significant decrease in ciliated cells and increase in hyperplastic goblet cells due to the single series of WTC dust exposures. The study also showed there was only a nominal non-significant decrease (6-11%) in WTC dust burden over a 1-yr period after the final exposure. These results provide support for our current hypothesis that exposure to WTC dusts caused changes in airway morphology/cell composition; such changes could, in turn, have led to potential alterations in the clearance/toxicities of other pollutants inhaled at Ground Zero in the critical initial 72-h period.


A Mobile System for Music Anamnesis and Receptive Music Therapy in the Personal Home.
Receptive music therapy is active hearing of music that is specifically selected to cause a certain effect on a person, such as pain reduction, mental opening, confrontation etc. This active, guided hearing could be helpful as a supporting ritual for patients at home and could extend traditional therapy. However, patients are often unable to select the music pieces that might be helpful for them in a current situation. We are suggesting a self-learning decision support system that allows a patient to answer questions on music anamnesis, is ready for inclusion into an electronic health record, and which enables a therapist to compile a therapeutic music program for the patient at home. Beyond this, the system also suggests appropriate music and duration of listening based on the patient's reported current mental state. In this paper, a concept for such a mobile system for receptive music therapy will be proposed.


Mitochondrial polymorphisms significantly reduce the risk of Parkinson disease.
Mitochondrial (mt) impairment, particularly within complex I of the electron transport system, has been implicated in the pathogenesis of Parkinson disease (PD). More than half of mitochondrially encoded polypeptides form part of the reduced nicotinamide adenine dinucleotide dehydrogenase (NADH) complex I enzyme. To test the hypothesis that mtDNA variation contributes to PD expression, we genotyped 10 single-nucleotide polymorphisms (SNPs) that define the European mtDNA haplogroups in 609 white patients with PD and 340 unaffected white control subjects. Overall, individuals classified as haplogroup J (odds ratio [OR] 0.55; 95% confidence interval [CI] 0.34-0.91; P=.02) or K (OR 0.52; 95% CI 0.30-0.90; P=.02) demonstrated a significant decrease in risk of PD versus individuals carrying the most common haplogroup, H. Furthermore, a specific SNP that defines these two haplogroups, 10398G, is strongly associated with this protective effect (OR 0.53; 95% CI 0.39-0.73; P=.0001). SNP 10398G causes a nonconservative amino acid change from threonine to alanine within the NADH dehydrogenase 3 (ND3) of complex I. After stratification by sex, this decrease in risk appeared stronger in women than in men (OR 0.43; 95% CI 0.27-0.71; P=.0009). In addition, SNP 9055A of ATP6 demonstrated a protective effect for women (OR 0.45; 95% CI 0.22-0.93; P=.03). Our results suggest that ND3 is an important factor in PD susceptibility among white individuals and could help explain the role of complex I in PD expression.


Accelerometers and pedometers: methodology and clinical application.
The relationship between physical activity and health varies considerably, partly due to the difficulty of assessing physical activity accurately. This review examines recent literature on the validation of movement sensors to assess habitual physical activity. Recommendations are given for the use of movement sensors during free-living conditions and methods of data analysis and interpretation are discussed. Recent progress in physical-activity research includes detailed comparative studies of different monitor brands. The move away from using linear-regression equations and the use of novel data-analysis strategies is increasing the accuracy with which energy expenditure can be estimated from accelerometry. New technologies, including the combination of accelerometry with the measurement of physiological parameters, have great potential for the increased accuracy of physical-activity assessment. Accelerometry is able to adequately assess physical activity and its association with health outcomes but currently methods have limited accuracy for the estimation of free-living energy expenditure. Pedometers provide an inexpensive overall measure of physical activity but are unable to assess intensity, frequency and duration of activity or to estimate energy expenditure. Interpretation of monitor output is best kept as close to the measurement domain as possible.


Optical detection of field cancerization in the buccal mucosa of patients with esophageal cancer.
Esophageal cancer is an increasingly common type of neoplasm with a very poor prognosis. This prognosis could improve with more early tumor detection. We have previously shown that we can use an optical spectroscopy to detect field cancerization in the buccal mucosa of patients with laryngeal cancer. The aim of this prospective study was to investigate whether we could detect field cancerization of buccal mucosa of patients with esophageal squamous cell carcinoma (ESCC) and esophageal adenocarcinoma (EAC). Optical measurements were performed in vivo using a novel optical technique: multidiameter single-fiber reflectance (MDSFR) spectroscopy. MDSFR spectra were acquired by a handheld probe incorporating three fiber diameters. Multiple absorption and scattering parameters that are related to the physiological and ultrastructural properties of the buccal mucosa were derived from these spectra. A linear discriminant analysis of the parameters was performed to create a combined biomarker σ to discriminate oncologic from non-oncologic patients. Twelve ESCC, 12 EAC, and 24 control patients were included in the study. The median value of our biomarker σ was significantly higher in patients with ESCC (2.07 [1.93-2.10]) than control patients (1.86 [1.73-1.95], p = 0.022). After cross-validation σ was able to identify ESCC patients with a sensitivity of 66.7% and a specificity of 70.8%. There were no significant differences between the EAC group and the control group. Field cancerization in the buccal mucosa can be detected using optical spectroscopy in ESCC patients. This may be the first step towards non-invasive ESCC cancer screening.


A Tale of Two Community Networks Program Centers: Operationalizing and Assessing CBPR Principles and Evaluating Partnership Outcomes.
Community Networks Program (CNP) centers are required to use a community-based participatory research (CBPR) approach within their specific priority communities. Not all communities are the same and unique contextual factors and collaborators' priorities shape each CBPR partnership. There are also established CBPR and community engagement (CE) principles shown to lead to quality CBPR in any community. However, operationalizing and assessing CBPR principles and partnership outcomes to understand the conditions and processes in CBPR that lead to achieving program and project level goals is relatively new in the science of CBPR. We sought to describe the development of surveys on adherence to and implementation of CBPR/CE principles at two CNP centers and examine commonalities and differences in program-versus project-level CBPR evaluation. A case study about the development and application of CBPR/CE principles for the Missouri CNP, Program for the Elimination of Cancer Disparities, and Minnesota CNP, Padres Informados/Jovenes Preparados, surveys was conducted to compare project versus program operationalization of principles. Survey participant demographics were provided by CNP. Specific domains found in CBPR/CE principles were identified and organized under an existing framework to establish a common ground. Operational definitions and the number of survey items were provided for each domain by CNP. There are distinct differences in operational definitions of CBPR/CE principles at the program and project levels of evaluation. However, commonalities support further research to develop standards for CBPR evaluation across partnerships and at the program and project levels.


Latin American and Caribbean countries' baseline clinical and policy guidelines for responding to intimate partner violence and sexual violence against women.
Violence against women is a global public health problem with negative effects on physical, mental, and reproductive health. The World Health Organization (WHO) has identified intimate partner violence (IPV) and sexual violence (SV) as major targets for prevention and amelioration and recently developed clinical and policy guidelines to assist healthcare providers. This project was undertaken to determine the 2013 baseline national policies and clinical guidelines on IPV and SV within the Latin American and Caribbean (LAC) region to identify strengths and gaps requiring action. Each Pan American Health Organization/World Health Organization Regional Office for the Americas (PAHO/WHO) country focal point was contacted to request their current national policy and clinical guidelines (protocol) on IPV/SV. We augmented this by searching the internet and the United Nations Women website. Each country's policy and clinical guideline (where available) was reviewed and entered into a scoring matrix based on WHO Clinical and Policy Guidelines. A total score for each heading and subheading was developed by adding positive responses to identify LAC regional strengths and gaps. We obtained 15 national policies and 12 national clinical guidelines (protocols) from a total of 18 countries ("response" rate 66.7%). National policies were comprehensive in terms of physical, emotional, and sexual violence and recommended good intersectoral collaboration. The greatest gap was in the training of health-care providers. National Guidelines for women-centered care for IPV/SV survivors were strong in the vital areas of privacy, confidentiality, danger assessment, safety planning, and supportive reactions to disclosure. The largest gaps noted were again in training healthcare professionals and strengthening monitoring and evaluation of services. Baseline measurement of policy and clinical guidelines for IPV/SV in LAC PAHO/WHO member countries at the time of issuing the 2013 WHO Clinical and Policy Guidelines reveals some important strengths, but also serious gaps that need to be addressed. The most pressing needs are for concerted training initiatives for healthcare providers and strengthening multisectoral monitoring and evaluation of services. A future evaluation of national policies, clinical guidelines, monitoring and evaluation will need to be conducted to measure the progress of the required scaling-up process.


Subpixel photometric stereo.
Conventional photometric stereo recovers one normal direction per pixel of the input image. This fundamentally limits the scale of recovered geometry to the resolution of the input image, and cannot model surfaces with subpixel geometric structures. In this paper, we propose a method to recover subpixel surface geometry by studying the relationship between the subpixel geometry and the reflectance properties of a surface. We first describe a generalized physically-based reflectance model that relates the distribution of surface normals inside each pixel area to its reflectance function. The distribution of surface normals can be computed from the reflectance functions recorded in photometric stereo images. A convexity measure of subpixel geometry structure is also recovered at each pixel, through an analysis of the shadowing attenuation. Then, we use the recovered distribution of surface normals and the surface convexity to infer subpixel geometric structures on a surface of homogeneous material by spatially arranging the normals among pixels at a higher resolution than that of the input image. Finally, we optimize the arrangement of normals using a combination of belief propagation and MCMC based on a minimum description length criterion on 3D textons over the surface. The experiments demonstrate the validity of our approach and show superior geometric resolution for the recovered surfaces.


Protein kinase C-epsilon is implicated in neurite outgrowth in differentiating human neuroblastoma cells.
A combination of basic fibroblast growth factor (bFGF) and insulin-like growth factor-I (IGF-I) or 16 nM 12-O-tetradecanoylphorbol-13-acetate (TPA) and serum induces human SH-SY5Y neuroblastoma cells to undergo differentiation and acquire a neuronal phenotype. Nerve growth factor (NGF) added to SH-SY5Y cells stably transfected with the NGF-receptor TRK-A (SH-SY5Y/trk) induces a similar differentiated phenotype. SH-SY5Y cells express protein kinase C (PKC)-alpha, PKC-beta I, PKC-epsilon, and PKC-zeta protein, and phorbol ester- or growth factor-induced differentiation results in a sustained activation of PKC. The specific PKC inhibitor GF 109203X blocked TPA- and bFGF-IGF-I-induced neurite outgrowth in wild-type SH-SY5Y cells and NGF-induced neurite outgrowth in SH-SY5Y/trk cells. When added to differentiated cells, GF 109203X caused rapid retraction of growth cone filopodia. In TPA- and bFGF-IGF-I-treated cells, addition of GF 109203X also blocked induced expression of growth associated protein-43 and neuropeptide tyrosine while the increase in expression of these two genes was only slightly affected by the inhibitor in NGF-treated SH-SY5Y/trk cells. Thus, a portion of the NGF-induced phenotypic changes appears not to be mediated via PKC-dependent signaling. A high concentration of TPA (1.6 microM) down regulated PKC-alpha and PKC-beta I almost completely and PKC-epsilon partially in wild-type SH-SY5Y and SH-SY5Y/trk cells. Cells with down-regulated PKC-alpha and PKC-beta I after 1.6 microM TPA treatment still differentiated with growth factors. In these cells, the PKC-epsilon level was restored, and the PKC-epsilon protein was enriched in the growth cones. The 1.6 microM TPA-induced down-regulation of PKC-epsilon was counteracted by bFGF and NGF but not by platelet-derived growth factor or IGF-I. These data indicate that PKC activity is vital for neurite formation, and that the cells can differentiate under conditions when PKC-alpha and PKC-beta I are extensively down regulated. The close correlation between differentiation and presence of PKC-epsilon protein suggests an important function for this isoform during this process.


A 4-item PRECISE-DAPT score for dual antiplatelet therapy duration decision-making.
The originally-proposed PRECISE-DAPT score is a 5-item risk score supporting decision-making for dual antiplatelet therapy1 duration after PCI. It is unknown if a simplified version of the score based on 4 factors (age, hemoglobin, creatinine clearance, prior bleeding), and lacking white-blood cell count, retains potential to guide DAPT duration. The 4-item PRECISE-DAPT was used to categorize 10,081 patients who were randomized to short (3-6 months) or long (12-24 months) DAPT regimen according to high (HBR defined by PRECISE-DAPT ≥25 points) or non-high bleeding risk (PRECISE-DAPT<25) status. Long treatment duration was associated with higher bleeding rates in HBR (ARD +2.22% [95% CI +0.53 to +3.90]) but not in non-HBR patients (ARD +0.25% [-0.14 to +0.64]; pint = 0.026), and associated with lower ischemic risks in non-HBR (ARD -1.44% [95% CI -2.56 to -0.31]), but not in HBR patients (ARD +1.16% [-1.91 to +4.22]; pint = 0.11). Only non-HBR patients experienced lower net clinical adverse events (NACE) with longer DAPT (pint = 0.043). A 4-item simplified version of the PRECISE-DAPT score retains the potential to categorize patients who benefit from prolonged DAPT without concomitant bleeding liability from those who do not.


Health of the Elderlies and Healthy Ageing: Challenge for Europe.
Population ageing is a major challenge for European Union (EU) society and economy, particularly for Italy, which is the oldest country in Europe. According to the World Health Organization, two-thirds of European citizens who have reached the retirement age suffer from at least two chronic conditions, with a strong pressure on healthcare systems. Moreover, EU countries already spend, on average, more than a quarter of their gross domestic product on social protection, above all pensions, health and long term care. The current financial crisis is putting a strain on this system. In this context, it becomes increasingly necessary to promote a healthy and independent ageing, by improving outcomes for patients and society while ensuring health systems sustainability. To this purpose a proactive approach to chronic diseases prevention (primary, secondary and tertiary) as well as an integrated healthcare approach and also patients' empowerment are required so as to make daily life more age-friendly. It is also necessary to share health and social best practices, adopt policies really effective against elderly social exclusion and strengthen older people participation in society. A joint effort of all key stakeholders is needed to create a society in which older people can play an active role.


A systematic review of criminal recidivism rates worldwide: 3-year update.
Background: Comparing recidivism rates between countries may provide useful information about the relative effectiveness of different criminal justice policies. A previous 2015 review identified criminal recidivism data for 18 countries and found little consistency in outcome definitions and time periods. We aimed to update recidivism rates in prisoners internationally. Methods: We conducted a systematic review of criminal recidivism rates in prisoners and followed PRISMA guidelines. Using three bibliographic indexes, we carried out non-country-specific and targeted searches for 50 countries with the largest total prison populations. We included reports and studies of released prisoners that reported re-arrest, reconviction and reincarceration rates. Meta-analysis was not possible due to multiple sources of heterogeneity. Results: We identified criminal recidivism information for 23 countries. Of the 50 countries with the largest prison populations, 10 reported recidivism rates for prisoners. The most commonly reported outcome was the 2-year reconviction rate. We were able to examine recidivism over different time periods for 11 countries and found that most reported small changes in official recidivism rates. Overall, for 2-year follow-up period, reported re-arrest rates were between 26% and 60%, reconviction rates ranged from 20% to 63%, and reimprisonment rates varied from 14 to 45%. Conclusions: Although some countries made efforts to improve reporting, recidivism rates are not comparable between countries. Criminal justice agencies should consider using reporting guidelines described here to update their data.


Optical phase space, Wigner representation, and invariant quality parameters.
Wigner's quasi probability and related functional and operator methods of quantum mechanics have recently played an important role in optics. We present an account of some of these developments. The symmetry structures underlying the ray and wave approaches to paraxial optics are explored in some detail, and the manner in which the Wigner phase-space representation captures the merits of both approaches is brought out. A fairly self-contained analysis of the second or intensity moments of general astigmatic partially coherent beams and of their behavior under transmission through astigmatic first-order optical systems is presented. Geometric representations of the intensity moments that render the quality parameters or polynomial invariants manifest are discussed, and the role of the optical uncertainty principle in assigning unbeatable physical bounds for these invariants is stressed. Measurement of the ten intensity moments of an astigmatic partially coherent beam is considered.


Is the effect of ill health on school achievement among Swedish adolescents gendered?
This study investigates why the relationship between health problems requiring hospitalization between the ages of 13 and 16 and school achievement (school grades in 9th grade) in Sweden was stronger for girls than for boys. We reviewed previous research on gender differences in subjective health, health care utilization and medical drug treatment to identify mechanisms responsible for this gendered effect. The relationship was analysed using retrospective observational data from several national full-population registers of individuals born in 1990 in Sweden (n = 115 196), and ordinary least squares techniques were used to test hypotheses. We found that girls had longer stays when hospitalized, which mediated 15% of the interaction effect. Variability in drug treatment between boys and girls did not explain the gendered effect of hospitalization. The main mediator of the gendered effect was instead differences in diagnoses between boys and girls. Girls' hospitalizations were more commonly related to mental and behavioural diagnoses, which have particularly detrimental effects on school achievement.


Inheritance of deleterious mutations at both BRCA1 and BRCA2 in an international sample of 32,295 women.
Most BRCA1 or BRCA2 mutation carriers have inherited a single (heterozygous) mutation. Transheterozygotes (TH) who have inherited deleterious mutations in both BRCA1 and BRCA2 are rare, and the consequences of transheterozygosity are poorly understood. From 32,295 female BRCA1/2 mutation carriers, we identified 93 TH (0.3 %). "Cases" were defined as TH, and "controls" were single mutations at BRCA1 (SH1) or BRCA2 (SH2). Matched SH1 "controls" carried a BRCA1 mutation found in the TH "case". Matched SH2 "controls" carried a BRCA2 mutation found in the TH "case". After matching the TH carriers with SH1 or SH2, 91 TH were matched to 9316 SH1, and 89 TH were matched to 3370 SH2. The majority of TH (45.2 %) involved the three common Jewish mutations. TH were more likely than SH1 and SH2 women to have been ever diagnosed with breast cancer (BC; p = 0.002). TH were more likely to be diagnosed with ovarian cancer (OC) than SH2 (p = 0.017), but not SH1. Age at BC diagnosis was the same in TH vs. SH1 (p = 0.231), but was on average 4.5 years younger in TH than in SH2 (p < 0.001). BC in TH was more likely to be estrogen receptor (ER) positive (p = 0.010) or progesterone receptor (PR) positive (p = 0.013) than in SH1, but less likely to be ER positive (p < 0.001) or PR positive (p = 0.012) than SH2. Among 15 tumors from TH patients, there was no clear pattern of loss of heterozygosity (LOH) for BRCA1 or BRCA2 in either BC or OC. Our observations suggest that clinical TH phenotypes resemble SH1. However, TH breast tumor marker characteristics are phenotypically intermediate to SH1 and SH2.


Clinically inexplicable frequent attenders in general practice.
This study was based in an isolated rural practice, and it identified 107 frequent attenders (1.95% of the practice population), who created five times the consultation workload of a similar group matched for age and sex. The general practitioners (GPs) classified 60 (56%) of these patients as attending for clinically inexplicable reasons. This subgroup had higher rates of long-term medication and hospital referral, and more free access to primary health care. The study also identified very high levels of kinship and relationship by marriage within this group (47%).


A National Survey on Moral Obligations in Critical Care.
Critical care clinicians routinely encounter ethically complex situations. Ethical conflicts sometimes arise from different perspectives regarding goals of care and moral obligations. These conflicts contribute to providers' moral distress and burnout and can erode trust between patients, families, and clinicians. To explore the most disturbing and most frequent types of ethically complex situations; compare clinicians' perceptions of their own, each other's, and shared moral obligations for providing quality care in these situations; and examine perceptions of communication and teamwork. A national, web-based survey was made available to members of the American Association of Critical-Care Nurses and the Society for Critical Care Medicine. The survey included rank order, rating, and open-ended questions. Nurses and physicians ranked similarly the most frequent and disturbing ethical situations encountered during critical care practice. Nurses and physicians rated similarly physicians' moral obligations, but their ratings of nurses' moral obligations differed, with physicians giving lower ratings. Physicians also were more likely than nurses to report higher levels of trust (90.6% vs 66.8%) and more satisfaction with team communication (81.3% vs 66.9%). Narrative comments revealed embedded, entwined, and sometimes divergent expectations about moral obligations, which interfered with effective teamwork during ethically complex situations. Teamwork in critical care would benefit from acknowledgment of and clear communication about role-specific, interdependent, and shared moral obligations. Opportunities for routine, team-based dialogue about ethical aspects of care and moral obligations could reduce role ambiguity and ethical conflicts.


Development of a novel FMRI compatible visual perception prototype battery to test older people with and without dementia.
Visuoperceptual deficits in dementia are common and can reduce quality of life. Testing of visuoperceptual function is often confounded by impairments in other cognitive domains and motor dysfunction. We aimed to develop, pilot, and test a novel visuocognitive prototype test battery which addressed these issues, suitable for both clinical and functional imaging use. We recruited 23 participants (14 with dementia, 6 of whom had extrapyramidal motor features, and 9 age-matched controls). The novel Newcastle visual perception prototype battery (NEVIP-B-Prototype) included angle, color, face, motion and form perception tasks, and an adapted response system. It allows for individualized task difficulties. Participants were tested outside and inside the 3T functional magnetic resonance imaging (fMRI) scanner. Functional magnetic resonance imaging data were analyzed using SPM8. All participants successfully completed the task inside and outside the scanner. Functional magnetic resonance imaging analysis showed activation regions corresponding well to the regional specializations of the visual association cortex. In both groups, there was significant activity in the ventral occipital-temporal region in the face and color tasks, whereas the motion task activated the V5 region. In the control group, the angle task activated the occipitoparietal cortex. Patients and controls showed similar levels of activation, except on the angle task for which occipitoparietal activation was lower in patients than controls. Distinct visuoperceptual functions can be tested in patients with dementia and extrapyramidal motor features when tests use individualized thresholds, adapted tasks, and specialized response systems.


Remote sensing for predicting potential habitats of Oncomelania hupensis in Hongze, Baima and Gaoyou lakes in Jiangsu province, China.
Political and health sector reforms, along with demographic, environmental and socio-economic transformations in the face of global warming, could cause the re-emergence of schistosomiasis in areas where transmission has been successfully interrupted and its emergence in previously non-endemic areas in China. In the present study, we used geographic information systems and remote sensing techniques to predict potential habitats of Oncomelania hupensis, the intermediate host snail of Schistosoma japonicum. Focussing on the Hongze, Baima and Gaoyou lakes in Jiangsu province in eastern China, we developed a model using the normalized difference vegetation index, a tasseled-cap transformed wetness index, and flooding areas to predict snail habitats at a small scale. Data were extracted from two Landsat images, one taken during a typical dry year and the other obtained three years later during a flooding event. An area of approximately 163.6 km2 was predicted as potential O. hupensis habitats around the three lakes, which accounts for 4.3% of the estimated snail habitats in China. In turn, these predicted snail habitats are risk areas for transmission of schistosomiasis, and hence illustrate the scale of the possible impact of climate change and other ecological transformations. The generated risk map can be used by health policy makers to guide mitigation policies targetting the possible spread of O. hupensis, and with the aim of containing the transmission of S. japonicum.


Controlling the self: the role of the dorsal frontomedian cortex in intentional inhibition.
Intentional inhibition refers to the suppression of ongoing behavior on the basis of internally-generated decisions. This ability to cancel planned actions at the last moment is thought to be critical for self-control and has been related to activation in a circumscribed region of the dorsal frontomedian cortex (dFMC). Preliminary theories of intentional inhibition were based on studies that exclusively examined the cancellation of motor responses, and consequently concluded that this region serves the suppression of motor output. Yet recent evidence suggests that the dFMC is also involved in inhibitory control over more abstract internal states such as emotions or desires that have no immediate behavioral output. In this review, we therefore wish to put forth a new integrative perspective on the role of the dFMC in human self-control. We will argue that by virtue of its anatomical location and functional connections, this area may subserve the disengagement from current urges and impulses, thus facilitating successful exertions of self-control across a wide range of contexts by overcoming a self-focused perspective. We will discuss the fit of this view of the dFMC with the existing literature, identify critical experimental determinants for engaging the dFMC in intentional inhibition, and outline promising perspectives for future research.


Comparison of Atmospheric Pressure Ionization Gas Chromatography-Triple Quadrupole Mass Spectrometry to Traditional High-Resolution Mass Spectrometry for the Identification and Quantification of Halogenated Dioxins and Furans.
The goal of this study was to qualify gas chromatography coupled to atmospheric pressure ionization tandem mass spectrometry (APGC-MS/MS) as a reliable and valid technique for analysis of halogenated dioxins and furans that could be used in place of more traditional gas chromatography coupled to high-resolution mass spectrometry (GC-HRMS) analysis. A direct comparison of the two instrumental techniques was performed. APGC-MS/MS system sensitivity was demonstrated to be on the single femtogram level. The APGC-MS/MS analysis also demonstrated method detection limits (MDLs) in both sediment and fish that were 2-18 times lower than those determined for the GC-HRMS. Inlet conditions were established to prevent issues with sample carry-over, due largely to the enhanced sensitivity of this technique. Additionally, this work utilized direct injection for sample introduction through the split/splittless inlet. Finally, quantification of both sediment and fish certified reference materials were directly compared between the APGC-MS/MS and GC-HRMS. The APGC-MS/MS performed similarly to, if not better than, the GC-HRMS instrument in the analysis of these samples. This data is intended to substantiate APGC-MS/MS as a comparable technique to GC-HRMS for the analysis of dioxins and furans.


Cardiac norepinephrine transporter protein expression is inversely correlated to chamber norepinephrine content.
The cardiac neuronal norepinephrine (NE) transporter (NET) in sympathetic neurons is responsible for uptake of released NE from the neuroeffector junction. The purpose of this study was to assess the chamber distribution of cardiac NET protein measured using [(3)H]nisoxetine binding in rat heart membranes and to correlate NE content to NET amount. In whole mounts of atria, NET was colocalized in nerve fibers with tyrosine hydroxylase (TH) immunoreactivity. NE content expressed as micrograms NE per gram tissue was lowest in the ventricles; however, NET binding was significantly higher in the left ventricle than the right ventricle and atria (P < 0.05), resulting in a significant negative correlation (r(2) = 0.922; P < 0.05) of NET to NE content. The neurotoxin 6-hydroxydopamine, an NET substrate, reduced NE content more in the ventricles than the atria, demonstrating functional significance of high ventricular NET binding. In summary, there is a ventricular predominance of NET binding that corresponds to a high NE reuptake capacity in the ventricles, yet negatively correlates to tissue NE content.


Light and electron microscopic immunocytochemistry of the same nerves from whole mount preparations.
A technique for performing correlated light and electron microscopic immunocytochemical studies on whole mount preparations has been developed using myenteric plexus from guinea pig small intestine as a model. With this method a structure containing a particular antigen can first be located by light microscopy and then examined with the electron microscope. Pieces of intestine pinned on balsa were incubated in oxygenated Krebs solution at 37 degrees C for 90-120 min and then fixed for 1 hr at room temperature in 4% formaldehyde, 0.05% glutaraldehyde, and 0.2% picric acid in 0.1 M sodium phosphate buffer, pH 7.4. The tissue was washed vigorously in several changes of 50% ethanol until the picric acid had been removed, stored overnight in phosphate buffer, and then exposed to 0.1% sodium cyanoborohydride in buffer for 30 min. Vasoactive intestinal peptide (VIP) was localized in separated layers containing myenteric plexus and longitudinal muscle using the peroxidase-antiperoxidase technique with imidazole intensification of the diaminobenzidine reaction product. At the light microscope level, tissue stained by this technique showed VIP-immunoreactive nerve cell bodies and processes throughout the thickness of the myenteric ganglia in numbers approximately equivalent to those seen in whole mounts processed by an established technique for the light microscopic demonstration of VIP, which does not involve exposure of tissue to glutaraldehyde. VIP-immunoreactive structures that were first identified at the light microscope level were subsequently examined at the electron microscope level. VIP-immunoreactive axon profiles were found to form synapses on both immunoreactive and nonimmunoreactive myenteric neurons. The fine structural appearance of the different cell types present in whole mount preparations prepared by this method was similar to that seen in conventionally fixed tissue, except that free and bound ribosomes were absent from the tissue processed for immunocytochemistry. The method described here is reliable and no more difficult than presently available methods for preembedding electron microscopic immunocytochemistry on sections. Its main advantage is that immunoreactive structures for ultrastructural study can be selected from the entire population of chemically identified nerves within a whole mount rather than from a smaller sample present within a section. This technique is applicable to other tissues that can be stained immunohistochemically in whole mounts. The fixation and penetration enhancement procedures can also be adapted for immunocytochemical studies on vibratome or frozen sections.


Observations on a recent increase in plagiocephaly without synostosis.
To verify and determine the cause of an increase in the referral of infants with plagiocephaly without synostosis (PWS) to a single tertiary craniofacial center. A chart review was performed for 269 infants with a diagnosis of PWS who presented to a single tertiary craniofacial center between 1979 and 1994. The pattern of referral for PWS was analyzed using both simple linear regression and time series regression analyses. In addition, the referral pattern for PWS was compared with that for infants seen at the same center who received a diagnosis of synostotic plagiocephaly. Changes in the distribution of several demographic, perinatal, and clinical variables during the study period were also assessed. Finally, in an effort to identify correlates of the risk of PWS developing, characteristics of patients who were Missouri residents and presented between 1992 and 1994 were evaluated and compared with those of the 1993 Missouri live birth cohort. The Cleft Palate and Craniofacial Deformities Institute, St Louis Children's Hospital, Washington University Medical Center. The average annual number of referrals to our center for PWS in the period 1992 to 1994 was more than sixfold greater than that for the preceding 13 years. There was a statistically significant increase in the annual number of referrals to our center during the 16-year study period. Moreover, there was evidence that the average annual increase in referrals was significantly greater during the last 3 years (1992 through 1994) of the study than in the first 13 years. This shift in the referral patterns is roughly contemporaneous with the American Academy of Pediatrics recommendations regarding infant sleep position. There was no evidence that either the mean number of referrals or the average annual increase in referrals for patients with synostosis changed during the study period. Among patients with PWS, the average age at presentation did not change during the study period. There were also no significant changes in the distribution of other demographic, perinatal, and clinical variables. When compared with the Missouri birth cohort, infants with PWS were significantly more likely to be boys and to have been delivered by forceps. There was also some evidence that patients with PWS were more likely to be born prematurely and to be products of multiple-gestation pregnancies. These associations were, however, of only borderline statistical significance. Referrals to our center for PWS increased markedly in 1992 relative to previous years. The temporal coincidence of this increase with the American Academy of Pediatrics recommendation to avoid the prone sleeping position, to reduce the risk of sudden infant death syndrome, suggests a possible causal relationship. If this association is causal, education regarding the need for head position rotation coupled with that for sudden infant death syndrome should obviate positional PWS.


The Role of 18F-FDG PET/CT in Staging and Prognostication of Mantle Cell Lymphoma: An Italian Multicentric Study.
Mantle cell lymphoma (MCL) is an aggressive lymphoma subtype with poor prognosis in which 18F-FDG-PET/CT role in treatment response evaluation and prediction of outcome is still unclear. The aim of this multicentric study was to investigate the role of 18F-FDG-PET/CT in staging MCL and the prognostic role of Deauville criteria (DC) in terms of progression-free survival (PFS) and overall survival (OS). We retrospectively enrolled 229 patients who underwent baseline and end-of-treatment (eot) 18F-FDG-PET/CT after first-line therapy. EotPET/CT scans were visually interpreted according to DC. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of PET/CT for evaluation of bone marrow (BM) were 27%, 100%, 100%, 48% and 57%, respectively. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of PET/CT for evaluation of the gastrointestinal (GI) tract were 60%, 99%, 93%, 90% and 91%, respectively. At a median follow-up of 40 months, relapse occurred in 104 cases and death in 49. EotPET/CT results using DC significantly correlated with PFS, not with OS. Instead, considering OS, only MIPI score was significantly correlated. In conclusion, we demonstrated that MCL is an FDG-avid lymphoma and 18F-FDG-PET/CT is a useful tool for staging purpose, showing good specificity for BM and GI evaluation, but suboptimal sensitivity. EotPET/CT result was the only independent significant prognostic factor that correlated with PFS.


Association of Bipolar Disorder With Major Adverse Cardiovascular Events: A Population-Based Historical Cohort Study.
This study aimed to assess the association of bipolar disorder (BD) with risk of major adverse cardiac events (MACEs) after adjusting for established cardiovascular disease (CVD) risk factors. We conducted a population-based historical cohort study using the Rochester Epidemiology Project. Patients older than 30 years with a clinical encounter from 1998 to 2000 with no prior MACE, atrial fibrillation, or heart failure were followed up through March 1, 2016. BD diagnosis was validated by chart review. Cox proportional hazards regression models were adjusted for established CVD risk factors, alcohol use disorder, other substance use disorders (SUDs), and major depressive disorder (MDD). The cohort included 288 individuals with BD (0.81%) and 35,326 individuals without BD as the reference group (Ref). Median (interquartile range) follow-up was 16.5 (14.6-17.5) years. A total of 5636 MACE events occurred (BD, 59; Ref, 5577). Survival analysis showed an association between BD and MACE (median event-free-survival rates: BD, 0.80; Ref, 0.86; log-rank p = .018). Multivariate regression adjusting for age and sex also yielded an association between BD and MACE (hazard ratio [HR] = 1.93; 95% confidence interval [CI] = 1.43-2.52; p < .001). The association remained significant after further adjusting for smoking, diabetes mellitus, hypertension, high-density lipoprotein cholesterol, and body mass index (HR = 1.66; 95% CI = 1.17-2.28; p = .006), and for alcohol use disorder, SUD, and MDD (HR = 1.56; 95% CI = 1.09-2.14; p = .010). In this study, BD was associated with an increased risk of MACE, which persisted after adjusting for established CVD risk factors, SUDs, and MDD. These results suggest that BD is an independent risk factor for major clinical cardiac disease outcomes.


The genome sequence of the facultative intracellular pathogen Brucella melitensis.
Brucella melitensis is a facultative intracellular bacterial pathogen that causes abortion in goats and sheep and Malta fever in humans. The genome of B. melitensis strain 16M was sequenced and found to contain 3,294,935 bp distributed over two circular chromosomes of 2,117,144 bp and 1,177,787 bp encoding 3,197 ORFs. By using the bioinformatics suite ERGO, 2,487 (78%) ORFs were assigned functions. The origins of replication of the two chromosomes are similar to those of other alpha-proteobacteria. Housekeeping genes, including those involved in DNA replication, transcription, translation, core metabolism, and cell wall biosynthesis, are distributed on both chromosomes. Type I, II, and III secretion systems are absent, but genes encoding sec-dependent, sec-independent, and flagella-specific type III, type IV, and type V secretion systems as well as adhesins, invasins, and hemolysins were identified. Several features of the B. melitensis genome are similar to those of the symbiotic Sinorhizobium meliloti.


Urban agriculture and Anopheles habitats in Dar es Salaam, Tanzania.
A cross-sectional survey of agricultural areas, combined with routinely monitored mosquito larval information, was conducted in urban Dar es Salaam, Tanzania, to investigate how agricultural and geographical features may influence the presence of Anopheles larvae. Data were integrated into a geographical information systems framework, and predictors of the presence of Anopheles larvae in farming areas were assessed using multivariate logistic regression with independent random effects. It was found that more than 5% of the study area (total size 16.8 km2) was used for farming in backyard gardens and larger open spaces. The proportion of habitats containing Anopheles larvae was 1.7 times higher in agricultural areas compared to other areas (95% confidence interval = 1.56-1.92). Significant geographic predictors of the presence of Anopheles larvae in gardens included location in lowland areas, proximity to river, and relatively impermeable soils. Agriculture-related predictors comprised specific seedbed types, mid-sized gardens, irrigation by wells, as well as cultivation of sugar cane or leafy vegetables. Negative predictors included small garden size, irrigation by tap water, rainfed production and cultivation of leguminous crops or fruit trees. Although there was an increased chance of finding Anopheles larvae in agricultural sites, it was found that breeding sites originated by urban agriculture account for less than a fifth of all breeding sites of malaria vectors in Dar es Salaam. It is suggested that strategies comprising an integrated malaria control effort in malaria-endemic African cities include participatory involvement of farmers by planting shade trees near larval habitats.


Detecting chronic kidney disease in population-based administrative databases using an algorithm of hospital encounter and physician claim codes.
Large, population-based administrative healthcare databases can be used to identify patients with chronic kidney disease (CKD) when serum creatinine laboratory results are unavailable. We examined the validity of algorithms that used combined hospital encounter and physician claims database codes for the detection of CKD in Ontario, Canada. We accrued 123,499 patients over the age of 65 from 2007 to 2010. All patients had a baseline serum creatinine value to estimate glomerular filtration rate (eGFR). We developed an algorithm of physician claims and hospital encounter codes to search administrative databases for the presence of CKD. We determined the sensitivity, specificity, positive and negative predictive values of this algorithm to detect our primary threshold of CKD, an eGFR <45 mL/min per 1.73 m² (15.4% of patients). We also assessed serum creatinine and eGFR values in patients with and without CKD codes (algorithm positive and negative, respectively). Our algorithm required evidence of at least one of eleven CKD codes and 7.7% of patients were algorithm positive. The sensitivity was 32.7% [95% confidence interval: (95% CI): 32.0 to 33.3%]. Sensitivity was lower in women compared to men (25.7 vs. 43.7%; p <0.001) and in the oldest age category (over 80 vs. 66 to 80; 28.4 vs. 37.6 %; p < 0.001). All specificities were over 94%. The positive and negative predictive values were 65.4% (95% CI: 64.4 to 66.3%) and 88.8% (95% CI: 88.6 to 89.0%), respectively. In algorithm positive patients, the median [interquartile range (IQR)] baseline serum creatinine value was 135 μmol/L (106 to 179 μmol/L) compared to 82 μmol/L (69 to 98 μmol/L) for algorithm negative patients. Corresponding eGFR values were 38 mL/min per 1.73 m² (26 to 51 mL/min per 1.73 m²) vs. 69 mL/min per 1.73 m² (56 to 82 mL/min per 1.73 m²), respectively. Patients with CKD as identified by our database algorithm had distinctly higher baseline serum creatinine values and lower eGFR values than those without such codes. However, because of limited sensitivity, the prevalence of CKD was underestimated.


Selective block of transient Ca channel current in mouse neuroblastoma cells by U-88779E.
Antioxidants and T-type Ca channel antagonists are neuroprotective during ischemia or other central nervous system traumas. U-88779E (1-[(4-chlorophenyl-phenyl)-methyl]-4-[(7-methoxy-5-isopropyl- 2,4,6-cycloheptatrien-1-one)-2-methyl]piperazine) has been shown to have both antioxidant activity and the ability to block Ca fluxes in cardiac microsomes. In this study, we examined the effect of U-88779E on Ca, Na and K channels in a neuronal cell line, N1E-115 cells. The drug blocked transient barium current (IBa) through low-threshold Ca channels (T-type) with little effect on other noninactivating IBa including the nifedipine-sensitive one. The drug at 20 microM reduced transient IBa at a constant rate, -7.2% of the control per min, and abolished the current within 15 min. This implies a continuous accumulation of the drug in cell membranes probably because of its high lipophilicity (log P = 7.003). U-88779E also blocked Na and K currents but at a rate about 8 times slower than that observed with transient IBa. Further studies on interactions of the drug with T-channels revealed that the drug had no effect on the shape of current-voltage curve, activation and inactivation kinetics, and steady-state activation curve. The drug, however, induced a hyperpolarizing shift in steady-state inactivation curve and became more potent under conditions where the channels in inactivated states prevail. These observations are consistent with the view that U-88779E has a higher affinity to T-type channels in inactivated states than in resting or open states.(ABSTRACT TRUNCATED AT 250 WORDS)


Disseminated and chronic Lyme borreliosis in Norway, 1995 - 2004.
Lyme borreliosis is the most common tickborne infection in Norway. All clinical manifestations of Lyme borreliosis other than erythema migrans are notifiable to Folkehelseinstituttet, the Norwegian Institute of Public Health. During the period 1995-2004 a total of 1506 cases of disseminated and chronic Lyme borreliosis were reported. Serological tests were the basis for laboratory diagnosis in almost all cases. The annual numbers of cases showed no clear trend over the period, but varied each year between 120 and 253 cases, with the highest number of cases reported in 2004. Seventy five per cent of cases with information on time of onset were in patients who fell ill during the months of June to October. There was marked geographical variation in reported incidence rates, with the highest rates reported from coastal counties in southern and central Norway. Fifty six per cent of the cases were in males and 44% in females. The highest incidence rate was found in children aged between 5 and 9 years. Neuroborreliosis was the most common clinical manifestation (71%), followed by arthritis/arthralgia (22%) and acrodermatitis chronica atrophicans (5%). Forty six per cent of patients were admitted to hospital. Prevention of borreliosis in Norway relies on measures to prevent tick bites, such as use of protective clothing and insect repellents, and early detection and removal of ticks. Antibiotics are generally not recommended for prophylaxis after tick bites in Norway.


RNA Silencing in Plants: Mechanisms, Technologies and Applications in Horticultural Crops.
Understanding the fundamental nature of a molecular process or a biological pathway is often a catalyst for the development of new technologies in biology. Indeed, studies from late 1990s to early 2000s have uncovered multiple overlapping but functionally distinct RNA silencing pathways in plants, including the posttranscriptional microRNA and small interfering RNA pathways and the transcriptional RNA-directed DNA methylation pathway. These findings have in turn been exploited for developing artificial RNA silencing technologies such as hairpin RNA, artificial microRNA, intrinsic direct repeat, 3' UTR inverted repeat, artificial trans-acting siRNA, and virus-induced gene silencing technologies. Some of these RNA silencing technologies, such as the hairpin RNA technology, have already been widely used for genetic improvement of crop plants in agriculture. For horticultural plants, RNA silencing technologies have been used to increase disease and pest resistance, alter plant architecture and flowering time, improve commercial traits of fruits and flowers, enhance nutritional values, remove toxic compounds and allergens, and develop high-value industrial products. In this article we aim to provide an overview of the RNA silencing pathways in plants, summarize the existing RNA silencing technologies, and review the current progress in applying these technologies for the improvement of agricultural crops particularly horticultural crops.


Indole-3-carbinol and ultraviolet B induce apoptosis of human melanoma cells via down-regulation of MITF.
We investigated the mechanism of indole-3-carbinol (13C)/ultraviolet B (UVB)-induced apoptosis using SK-MEL-2 and SK-MEL-5 human melanoma cells. 13C/UVB significantly reduced the viability of SK-MEL-2 cells, whereas it had little influence on SK-MEL-5 cells. Correspondingly, cell cycle analysis showed that 13C/UVB induced a clear increase in the sub-G0/G1 phase in SK-MEL-2 cells. Furthermore, 13C/UVB activated caspase-9, caspase-8, caspase-3, and Bid and caused the cleavage of poly(ADP-ribose) polymerase (PARP) in SK-MEL-2 cells. In contrast, 13C/UVB showed no effects on the apoptotic signaling pathways in SK-MEL-5 cells. Moreover, we found that 13C down-regulated the microphthalmia-associated transcription factor (MITF) in SK-MEL-2 cells, but not in SK-MEL-5 cells. Next, to investigate the involvement of MITF in 13C/UVB-induced apoptosis, MITF silencing was conducted using small interfering RNA (siRNA) for MITF in SK-MEL-5 cells. Interestingly, 13C/UVB dramatically decreased the viability of MITF-down-regulated SK-MEL-5 cells. These results indicate that MITF plays a critical role in melanoma cell survival.


Relevance of distinct monocyte subsets to clinical course of ischemic stroke patients.
The most common strategy for treating patients with acute ischemic stroke is thrombolytic therapy, though only a few patients receive benefits because of the narrow time window. Inflammation occurring in the central nervous system (CNS) in association with ischemia is caused by immune cells including monocytes and involved in lesion expansion. If the specific roles of monocyte subsets in stroke can be revealed, they may become an effective target for new treatment strategies. We performed immunological examinations of 36 consecutive ischemic stroke patients within 2 days of onset and compared the results with 24 age-matched patients with degenerative disorders. The stroke patients were repeatedly tested for the proportions of monocyte subsets in blood, and serum levels of pro- and anti-inflammatory cytokines immediately after admission, on days 3-7 and 12-16 after stroke onset, and on the day of discharge. In addition, immunological measurements were analyzed for relationships to stroke subtypes and complications, including progressive infarction (PI) and stroke-associated infection (SAI). Monocyte count was significantly increased from 0-16 days after stroke as compared to the controls (p<0.05). CD14(high)CD16(-) classical and CD14(high)CD16(+) intermediate monocytes were significantly increased from 0-7 and 3-16 days after stroke, respectively (p<0.05), whereas CD14 (dim)CD16(high) non-classical monocytes were decreased from 0-7 days (p<0.05). Cardioembolic infarction was associated with a persistent increase in intermediate monocytes. Furthermore, intermediate monocytes were significantly increased in patients with PI (p<0.05), while non-classical monocytes were decreased in those with SAI (p<0.05). IL-17A levels were positively correlated with monocyte count (r=0.485, p=0.012) as well as the percentage of non-classical monocytes (r=0.423, p=0.028), and negatively with that of classical monocytes (r=-0.51, p=0.007) during days 12-16. Our findings suggest that CD14(high)CD16(+) intermediate monocytes have a role in CNS tissue damage during acute and subacute phases in ischemic stroke especially in relation to cardioembolism.


An N-terminally acetylated Arf-like GTPase is localised to lysosomes and affects their motility.
Small GTPases of the Arf and Rab families play key roles in the function of subcellular organelles. Each GTPase is usually found on only one compartment and, hence, they confer organelle specificity to many intracellular processes. However, there has so far been little evidence for specific GTPases present on lysosomes. Here, we report that two closely related human Arf-like GTPases, Arl8a and Arl8b (also known as Arl10b/c and Gie1/2), localise to lysosomes in mammalian cells, with the single homologue in Drosophila cells having a similar location. Conventionally, membrane binding of Arf and Arl proteins is mediated by both an N-terminal myristoyl group and an N-terminal amphipathic helix that is inserted into the lipid bilayer upon activation of the GTPase. Arl8a and Arl8b do not have N-terminal myristoylation sites, and we find that Arl8b is instead N-terminally acetylated, and an acetylated methionine is necessary for its lysosomal localization. Overexpression of Arl8a or Arl8b results in a microtubule-dependent redistribution of lysosomes towards the cell periphery. Live cell imaging shows that lysosomes move more frequently both toward and away from the cell periphery, suggesting a role for Arl8a and Arl8b as positive regulators of lysosomal transport.


Sensitivity of pituitary gonadotropes to hyperglycemia leads to epigenetic aberrations and reduced follicle-stimulating hormone levels.
The connection between metabolism and reproductive function is well recognized, and we hypothesized that the pituitary gonadotropes, which produce luteinizing hormone and follicle-stimulating hormone (FSH), mediate some of the effects directly via insulin-independent glucose transporters, which allow continued glucose metabolism during hyperglycemia. We found that glucose transporter 1 is the predominant glucose transporter in primary gonadotropes and a gonadotrope precursor-derived cell line, and both are responsive to culture in high glucose; moreover, metabolite levels were altered in the cell line. Several of the affected metabolites are cofactors for chromatin-modifying enzymes, and in the gonadotrope precursor-derived cell line, we recorded global changes in histone acetylation and methylation, decreased DNA methylation, and increased hydroxymethylation, some of which did not revert to basal levels after cells were returned to normal glucose. Despite this weakening of epigenetic-mediated repression seen in the model cell line, FSH β-subunit ( Fshb) mRNA levels in primary gonadotropes were significantly reduced, apparently due in part to increased autocrine/paracrine effects of inhibin. However, unlike thioredoxin interacting protein and inhibin subunit α, Fshb mRNA levels did not recover after the return of cells to normal glucose. The effect on Fshb expression was also seen in 2 hyperglycemic mouse models, and levels of circulating FSH, required for follicle growth and development, were reduced. Thus, hyperglycemia seems to target the pituitary gonadotropes directly, and the likely extensive epigenetic changes are sensed acutely by Fshb. This scenario would explain clinical findings in which, even after restoration of optimal blood glucose levels, fertility often remains adversely affected. However, the relative accessibility of the pituitary provides a possible target for treatment, particularly crucial in the young in which hyperglycemia is increasingly common and fertility most relevant.-Feldman, A., Saleh, A., Pnueli, L., Qiao, S., Shlomi, T., Boehm, U., Melamed, P. Sensitivity of pituitary gonadotropes to hyperglycemia leads to epigenetic aberrations and reduced follicle-stimulating hormone levels.


Perceived barriers to accessing Female Community Health Volunteers' (FCHV) services among ethnic minority women in Nepal: A qualitative study.
Despite the efforts of community health workers to increase access to healthcare among ethnic minority groups in low- and -middle income countries, members of ethnic minorities are less likely than women from other ethnic groups to use maternal and child healthcare services. However, much less is known about the factors that limit access of ethnic minorities to healthcare services, including the services of community health workers in Nepal, who are known as Female Community Health Volunteers (FCHVs). To address this issue, we conducted a qualitative study to explore perceived barriers to accessing maternal and child healthcare services among ethnic minority groups in two different geographical locations (the hill and Terai regions- flatland bordering south India) with varying degrees of access to local healthcare centres. Between April 2014 and September 2014, semi-structured interviews were conducted with twenty FCHVs, 26 women service users and 11 paid local health workers. In addition, 15 FCHVs participated in four focus group discussions. A thematic analysis of the data identified five major themes underlying barriers to accessing available maternal and child healthcare services by ethnic minority groups such as Dalits, Madhesi, Muslim, Chepang and Tamang. These themes include: a) lack of knowledge among service users; b) lack of trust in volunteers; c) traditional beliefs and healthcare practices; d) low decision-making power of women; and e) perceived indignities experienced when using health centres. We conclude that community health programmes should focus on increasing awareness of healthcare services among ethnic minority groups, and the programmes should involve family members (husband and mothers-in-law) and traditional health practitioners. Both the FCHVs and local healthcare providers should be trained to communicate effectively in order to deliver respectful care among ethnic minorities if we want to achieve universal healthcare coverage for maternal and child health in low- and -middle income countries.


Comparative analysis of the combined petrosal and the pretemporal transcavernous anterior petrosal approach to the petroclival region.
The combined petrosal (CP) approach has been traditionally used to resect petroclival meningioma (PCM). The pretemporal transcavernous anterior petrosal (PTAP) approach has emerged as an alternative. A quantitative comparison of both approaches has not been made. This anatomical study compared the surgical corridors afforded by both approaches and identified key elements of the approach selection process. Twelve cadaveric specimens were dissected, and 10 were used for morphometric analysis. Groups A and B (n = 5 in each) underwent the CP and PTAP approaches, respectively. The area of drilled clivus, lengths of cranial nerves (CNs) II-X, length of posterior circulation vessels, surgical area of exposure of the brainstem, and angles of attack anterior and posterior to a common target were measured and compared. The area of drilled clivus was significantly greater in group A than group B (mean ± SD 88.7 ± 17.1 mm2 vs 48.4 ± 17.9 mm2, p < 0.01). Longer segments of ipsilateral CN IV (52.4 ± 2.33 mm vs 46.5 ± 3.71 mm, p < 0.02), CN IX, and CN X (9.91 ± 3.21 mm vs 0.00 ± 0.00 mm, p < 0.01) were exposed in group A than group B. Shorter portions of CN II (9.31 ± 1.28 mm vs 17.6 ± 6.89 mm, p < 0.02) and V1 (26.9 ± 4.62 mm vs 32.4 ± 1.93 mm, p < 0.03) were exposed in group A than group B. Longer segments of ipsilateral superior cerebellar artery (SCA) were exposed in group A than group B (36.0 ± 4.91 mm vs 25.8 ± 3.55 mm, p < 0.02), but there was less exposure of contralateral SCA (0.00 ± 0.00 mm vs 7.95 ± 3.33 mm, p < 0.01). There was no statistically significant difference between groups with regard to the combined area of the exposed cerebral peduncles and pons (p = 0.75). Although exposure of the medulla was limited, group A had significantly greater exposure of the medulla than group B (p < 0.01). Finally, group A had a smaller anterior angle of attack than group B (24.1° ± 5.62° vs 34.8° ± 7.51°, p < 0.03). This is the first study to quantitatively identify the advantages and limitations of the CP and PTAP approaches from an anatomical perspective. Understanding these data will aid in designing maximally effective yet minimally invasive approaches to PCM.


Combining tumor genome simulation with crowdsourcing to benchmark somatic single-nucleotide-variant detection.
The detection of somatic mutations from cancer genome sequences is key to understanding the genetic basis of disease progression, patient survival and response to therapy. Benchmarking is needed for tool assessment and improvement but is complicated by a lack of gold standards, by extensive resource requirements and by difficulties in sharing personal genomic information. To resolve these issues, we launched the ICGC-TCGA DREAM Somatic Mutation Calling Challenge, a crowdsourced benchmark of somatic mutation detection algorithms. Here we report the BAMSurgeon tool for simulating cancer genomes and the results of 248 analyses of three in silico tumors created with it. Different algorithms exhibit characteristic error profiles, and, intriguingly, false positives show a trinucleotide profile very similar to one found in human tumors. Although the three simulated tumors differ in sequence contamination (deviation from normal cell sequence) and in subclonality, an ensemble of pipelines outperforms the best individual pipeline in all cases. BAMSurgeon is available at https://github.com/adamewing/bamsurgeon/.


Neurophysiological evidence of corollary discharge function during vocalization in psychotic patients and their nonpsychotic first-degree relatives.
Predictions about sensations resulting from motor acts are instantiated through neural mechanisms such as the corollary discharge. With each action, the corollary discharge provides an unconscious comparison between predicted and actual sensations resulting from the action; closer matches result in greater suppression of sensation. This mechanism is disrupted in schizophrenia (SZ) and may contribute to, or reflect a failure to, distinguish self- from externally generated experiences, a hallmark of psychosis. We asked whether disruption is specific to SZ or is seen in other psychotic illnesses and in first-degree relatives of psychotic patients. Corollary discharge function was assessed in SZ patients (n = 30), schizoaffective (SA) patients (n = 19), bipolar patients with a history of psychosis (BPP; n = 39), nonpsychotic relatives of SZ (n = 30), SA (n = 23), and BPP (n = 50) patients, and healthy controls (n = 43). The N1 component of the event-related potential, reflecting auditory cortical responses to sounds, was elicited by speech sound onset as subjects talked and later when they listened to a recording of those sounds. N1 was suppressed during talking compared to N1 during listening, consistent with the suppressive action of the corollary discharge mechanism. Suppression was significantly reduced in SZ and BPP patients, with a similar trend in the smaller SA group. Patient groups did not differ, and unaffected relatives did not differ from controls or probands. The failure to monitor sensations resulting from self-generated actions, implicating corollary discharge dysfunction, may be a common feature across affective and nonaffective psychosis. Data from unaffected family members do not indicate that this is a marker of psychosis risk.


Commonalities of cerclage-related genitourinary fistulas.
The purpose of this review was to identify common features of cerclage-related genitourinary fistulas. A retrospective case series of cerclage-related fistulas was performed. The results were then pooled with available published case reports to identify common factors in these cases. Fistula location, surgical technique, and previous surgical and obstetrical histories were examined. From 2008 to 2011, 5 women were referred with vesicovaginal fistulas after cerclage. Within this series, 4 had prior cervical procedures, 3 had previous cesarean deliveries, and 4 had a McDonald cerclage in the antecedent pregnancy. When pooled with available data in 7 published case reports on cerclage-related fistulas, all 12 fistulas occurred in patients with prior histories of cervical procedures or cesarean deliveries. Specifically, of the 12 patients, 8 [66.7%; 95% confidence interval (CI), 0.39-0.86] had at least 1 prior cerclage and 10 (83.3%; 95% CI, 0.54-0.97) had at least 1 prior cervical surgery. When reported, 81.8% (95% CI, 0.51-0.96) had the McDonald technique used for placement of the current cerclage. The isolated and pooled findings suggest previous cerclage, any previous cervical procedures, and use of the McDonald technique are common factors in cases of cerclage-related genitourinary fistulas. This information may be useful when evaluating and counseling patients.


Effective or just practical? An evaluation of an online postgraduate module on evidence-based medicine (EBM).
Teaching the steps of evidence-based medicine (EBM) to undergraduate as well as postgraduate health care professionals is crucial for implementation of effective, beneficial health care practices and abandonment of ineffective, harmful ones. Stellenbosch University in Cape Town, South Africa, offers a 12-week, completely online module on EBM within the Family Medicine division, to medical specialists in their first year of training. The aim of this study was to formatively evaluate this module; assessing both the mode of delivery; as well as the perceived effectiveness and usefulness thereof. We used mixed methods to evaluate this module: A document review to assess whether the content of the module reflects important EBM competencies; a survey of the students to determine their experiences of the module; and semi-structured interviews with the tutors to explore their perspectives of the module. Ethics approval was obtained. The document review indicated that EBM competencies were covered adequately, although critical appraisal only focused on randomised controlled trials and guidelines. Students had a positive attitude towards the module, but felt that they needed more support from the tutors. Tutors felt that students engaged actively in discussions, but experienced difficulties with understanding certain concepts of EBM. Furthermore, they felt that it was challenging explaining these via the online learning platform and saw the need to incorporate more advanced technology to better connect with the students. In their view the key to successful learning of EBM was to keep it relevant and applicable to everyday practice. Tutors also felt that an online module on EBM was advantageous, since doctors from all over the world were able to participate. Our study has shown that the online module on EBM was effective in increasing EBM knowledge and skills of postgraduate students and was well received by both students and tutors. Students and tutors experienced generic challenges that accompany any educational intervention of EBM (e.g. understanding difficult concepts), but in addition had to deal with challenges unique to the online learning environment. Teachers of EBM should acknowledge these so as to enhance and successfully implement EBM teaching and learning for all students.


Working memory in middle-aged males: age-related brain activation changes and cognitive fatigue effects.
We examined the effects of aging and cognitive fatigue on working memory (WM) related brain activation using functional magnetic resonance imaging. Age-related differences were investigated in 13 young and 16 middle-aged male school teachers. Cognitive fatigue was induced by sustained performance on cognitively demanding tasks (compared to a control condition). Results showed a main effect of age on left dorsolateral prefrontal and superior parietal cortex activation during WM encoding; greater activation was evident in middle-aged than young adults regardless of WM load or fatigue condition. An interaction effect was found in the dorsomedial prefrontal cortex (DMPFC); WM load-dependent activation was elevated in middle-aged compared to young in the control condition, but did not differ in the fatigue condition due to a reduction in activation in middle-aged in contrast to an increase in activation in the young group. These findings demonstrate age-related activation differences and differential effects of fatigue on activation in young and middle-aged adults.


Kant on biological teleology: Towards a two-level interpretation.
According to Kant's Critique of the power of judgment, teleological considerations are unavoidable for conceptualizing organisms. Does this mean that teleology is more than merely heuristic? Kant stresses the regulative status of teleological attributions, but sometimes he seems to treat teleology as a constitutive condition for biology. To clarify this issue, the concept of natural purpose and its role for biology are examined. I suggest that the concept serves an identificatory function: it singles out objects as natural purposes, whereby the special science of biology is constituted. This relative constitutivity of teleology is explicated by means of a distinction of levels: on the object level of biological science, teleology is taken as constitutive, though it is merely regulative on the philosophical meta level. This distinction also concerns the place of Aristotelian teleology in Kant: on the object level, the Aristotelian view is accepted, whereas on the meta level, an agnostic stance is taken concerning teleology.


Bacterial community structure and dissolved organic matter in repeatedly flooded subsurface karst water pools.
Bacterial diversity, community assembly, and the composition of the dissolved organic matter (DOM) were studied in three temporary subsurface karst pools with different flooding regimes. We tested the hypothesis that microorganisms introduced to the pools during floods faced environmental filtering toward a 'typical' karst water community, and we investigated whether DOM composition was related to floodings and the residence time of water in stagnant pools. As predicted, longer water residence consistently led to a decline of bacterial diversity. The microbial assemblages in the influx water harbored more 'exotic' lineages with large distances to known genotypes, yet these initial communities already appeared to be shaped by selective processes. β-Proteobacterial operational taxonomic units (OTUs) closely related to microbes from subsurface or surface aquatic environments were mainly responsible for the clustering of samples according to water residence time in the pools. By contrast, several Cytophagaceae and Flavobacteriaceae OTUs were related to different floodings, which were also the main determinants of DOM composition. A subset of compounds distinguishable by molecular mass and O/C content were characteristic for individual floods. Moreover, there was a transformation of DOM in stagnant pools toward smaller and more aromatic compounds, potentially also reflecting microbial utilization.


Australian midwives and provision of nutrition education during pregnancy: A cross sectional survey of nutrition knowledge, attitudes, and confidence.
Maternal nutrition during pregnancy affects the health of the mother and the baby. Midwives are ideally placed to provide nutrition education to pregnant women. There is limited published research evidence of Australian midwives' nutrition knowledge, attitudes and confidence. To investigate Australian midwives' nutrition knowledge, attitudes and confidence in providing nutrition education during pregnancy. Members of the Australian College of Midwives (n=4770) were sent an invitation email to participate in a web-based survey, followed by two reminders. The completion rate was 6.9% (329 of 4770). The majority (86.6% and 75.7%, respectively) highly rated the importance of nutrition during pregnancy and the significance of their role in nutrition education. Midwives' nutrition knowledge was inadequate in several areas such as weight gain, dairy serves and iodine requirements (73.3%, 73.2% and 79.9% incorrect responses, respectively). The level of confidence in discussing general and specific nutrition issues ranged mostly from moderate to low. The majority of the midwives (93%) provided nutrition advice to pregnant women. This advice was mostly described as 'general' and focused on general nutrition topics. Only half of the midwives reported receiving nutrition education during midwifery education (51.1%) or after registration (54.1%). Australian midwives' attitudes towards nutrition during pregnancy and their role in educating pregnant women about it were positive but their knowledge and confidence did not align with these attitudes. This could be due to minimal nutrition education during midwifery education or during practice. Continued education to improve midwives' nutrition knowledge and confidence is essential.


Magnetic micro- and nanoparticle mediated activation of mechanosensitive ion channels.
Most cells are known to respond to mechanical cues, which initiate biochemical signalling pathways and play a role in cell membrane electrodynamics. These cues can be transduced either via direct activation of mechanosensitive (MS) ion channels or through deformation of the cell membrane and cytoskeleton. Investigation of the function and role of these ion channels is a fertile area of research and studies aimed at characterizing and understanding the mechanoactive regions of these channels and how they interact with the cytoskeleton are fundamental to discovering the specific role that mechanical cues play in cells. In this review, we will focus on novel techniques, which use magnetic micro- and nanoparticles coupled to external applied magnetic fields for activating and investigating MS ion channels and cytoskeletal mechanics.


School closures during COVID-19: an overview of systematic reviews.
To assess the benefits and drawbacks of school closures and in-school mitigations during the COVID-19 pandemic. Overview of systematic reviews (SRs). We searched six databases and additional resources on 29 July 2022: MEDLINE, Embase, Google Scholar, Cochrane Library, COVID-END inventory of evidence synthesis, and Epistemonikos. We selected SRs written in English that answered at least one of four specific questions concerning the efficacy and drawbacks of school closures. Their primary studies were conducted in primary and secondary schools, including pupils aged 5-18. Interventions included school closures or mitigations (such as mask usage) introduced in schools. We used AMSTAR 2 to assess confidence in the included SRs, and GRADE was used to assess certainty of evidence. We performed a narrative synthesis of the results, prioritising higher-quality SRs, those which performed GRADE assessments and those with more unique primary studies. We also assessed the overlap between primary studies included in the SRs. Our framework for summarising outcome data was guided by the following questions: (1) What is the impact of school closures on COVID-19 transmission, morbidity or mortality in the community? (2) What is the impact of COVID-19 school closures on mental health (eg, anxiety), physical health (eg, obesity, domestic violence, sleep) and learning/achievement of primary and secondary pupils? (3) What is the impact of mitigations in schools on COVID-19 transmission, morbidity or mortality in the community? and (4) What is the impact of COVID-19 mitigations in schools on mental health, physical health and learning/achievement of primary and secondary pupils? We identified 578 reports, 26 of which were included. One SR was of high confidence, 0 moderate, 10 low and 15 critically low confidence. We identified 132 unique primary studies on the effects of school closures on transmission/morbidity/mortality, 123 on learning, 164 on mental health, 22 on physical health, 16 on sleep, 7 on domestic violence and 69 on effects of in-school mitigations on transmission/morbidity/mortality.Both school closures and in-school mitigations were associated with reduced COVID-19 transmission, morbidity and mortality in the community. School closures were also associated with reduced learning, increased anxiety and increased obesity in pupils. We found no SRs that assessed potential drawbacks of in-school mitigations on pupils. The certainty of evidence according to GRADE was mostly very low. School closures during COVID-19 had both positive and negative impacts. We found a large number of SRs and primary studies. However, confidence in the SRs was mostly low to very low, and the certainty of evidence was also mostly very low. We found no SRs assessing the potential drawbacks of in-school mitigations on children, which could be addressed moving forward. This overview provides evidence that could inform policy makers on school closures during future potential waves of COVID-19.


Dyspnoea as a predictor of cause-specific heart/lung disease mortality in Bangladesh: a prospective cohort study.
The spectrum of mortality outcomes by cause in populations with/without dyspnoea has not been determined. The study aimed to evaluate whether dyspnoea, a symptom, predicts cause-specific mortality differences between groups. The hypothesis was that diseases that result in chronic dyspnoea, those originating from the heart and lungs, would preferentially result in heart and lung disease mortality in those with baseline dyspnoea (relative to no dyspnoea) when followed over time. A population-based sample of 11 533 Bangladeshis was recruited and followed for 11-12 years and cause-specific mortality evaluated in those with and without baseline dyspnoea. Dyspnoea was ascertained by trained physicians. The cause of death was determined by verbal autopsy. Kaplan-Meier survival curves, the Fine-Gray competing risk hazards model and logistic regression models were used to determine group differences in cause-specific mortality. Compared to those not reporting dyspnoea at baseline, the adjusted HRs were 6.4 (3.8 to 10.7), 9.3 (3.9 to 22.3), 1.8 (1.2 to 2.8), 2.2 (1.0 to 5.1) and 2.8 (1.3 to 6.2) for greater risk of dying from chronic obstructive pulmonary disease (COPD), asthma, heart disease, tuberculosis and lung cancer, respectively. In contrast, there was a similar risk of dying from stroke, cancer (excluding lung), liver disease, accidents and other (miscellaneous causes) between the dyspnoeic and non-dyspnoeic groups. In addition, the HR was 2.1 (1.7 to 2.5) for greater all-cause mortality in those with baseline dyspnoea versus no dyspnoea. Dyspnoea, ascertained by a single question with binary response, predicts heart and lung disease mortality. Individuals reporting dyspnoea were twofold to ninefold more likely to die of diseases that involve the heart and/or lungs relative to the non-dyspnoeic individuals. Therefore, in those with chronic dyspnoea, workup to look for the five common dyspnoeic diseases resulting in increased mortality (COPD, asthma, heart disease, tuberculosis and lung cancer), all treatable, should reduce mortality and improve the public health.


Changing cultural attitudes towards female genital cutting.
As globalization brings people with incompatible attitudes into contact, cultural conflicts inevitably arise. Little is known about how to mitigate conflict and about how the conflicts that occur can shape the cultural evolution of the groups involved. Female genital cutting is a prominent example. Governments and international agencies have promoted the abandonment of cutting for decades, but the practice remains widespread with associated health risks for millions of girls and women. In their efforts to end cutting, international agents have often adopted the view that cutting is locally pervasive and entrenched. This implies the need to introduce values and expectations from outside the local culture. Members of the target society may view such interventions as unwelcome intrusions, and campaigns promoting abandonment have sometimes led to backlash as they struggle to reconcile cultural tolerance with the conviction that cutting violates universal human rights. Cutting, however, is not necessarily locally pervasive and entrenched. We designed experiments on cultural change that exploited the existence of conflicting attitudes within cutting societies. We produced four entertaining movies that served as experimental treatments in two experiments in Sudan, and we developed an implicit association test to unobtrusively measure attitudes about cutting. The movies depart from the view that cutting is locally pervasive by dramatizing members of an extended family as they confront each other with divergent views about whether the family should continue cutting. The movies significantly improved attitudes towards girls who remain uncut, with one in particular having a relatively persistent effect. These results show that using entertainment to dramatize locally discordant views can provide a basis for applied cultural evolution without accentuating intercultural divisions.


The active form of the norovirus RNA-dependent RNA polymerase is a homodimer with cooperative activity.
Norovirus (NV) is a leading cause of gastroenteritis worldwide and a major public health concern. So far, the replication strategy of NV remains poorly understood, mainly because of the lack of a cell system to cultivate the virus. In this study, the function and the structure of a key viral enzyme of replication, the RNA-dependent RNA polymerase (RdRp, NS7), was examined. The overall structure of the NV NS7 RdRp was determined by X-ray crystallography to a 2.3 A (0.23 nm) resolution (PDB ID 2B43), displaying a right-hand fold typical of the template-dependent polynucleotide polymerases. Biochemical analysis evidenced that NV NS7 RdRp is active as a homodimer, with an apparent K(d) of 0.649 microM and a positive cooperativity (Hill coefficient n(H)=1.86). Crystals of the NV NS7 homodimer displayed lattices containing dimeric arrangements with high shape complementarity statistics. This experimental data on the structure and function of the NV RdRp may set the cornerstone for the development of polymerase inhibitors to control the infection with NV, a medically relevant pathogen.


Why verifying diagnostic decisions with a checklist can help: insights from eye tracking.
Making a diagnosis involves ratifying or verifying a proposed answer. Formalizing this verification process with checklists, which highlight key variables involved in the diagnostic decision, is often advocated. However, the mechanisms by which a checklist might allow clinicians to improve their verification process have not been well studied. We hypothesize that using a checklist to verify diagnostic decisions enhances analytic scrutiny of key variables, thereby improving clinicians' ability to find and fix mistakes. We asked 16 participants to verify their interpretation of 12 electrocardiograms, randomly assigning half to be verified with a checklist and half with an analytic prompt. While participants were verifying their interpretation, we tracked their eye movements. We analyzed these eye movements using a series of eye tracking variables theoretically linked to analytic scrutiny of key variables. We found that more errors were corrected using a checklist compared to an analytic prompt (.27 ± .53 errors per ECG vs. .04 ± .43, F 1,15 = 8.1, p = .01, η (2) = .20). Checklist use was associated with enhanced analytic scrutiny in all eye tracking measures assessed (F 6,10 = 6.0, p = .02). In this experiment, using a key variable checklist to verify diagnostic decisions improved error detection. This benefit was associated with enhanced analytic scrutiny of those key variables as measured by eye tracking.


Conservative management of nontubal ectopic pregnancies.
To report successful conservative management of nontubal ectopic pregnancies. Retrospective case series. University tertiary-care hospital. Sixty-four women with diagnosis of nontubal ectopic pregnancies (cervical, cornual, and cesarean section scar) were treated with minimally invasive procedures. Systemic methotrexate alone or combined with ultrasound-guided fetal intracardiac injection of potassium chloride. Success of the treatment, preservation of the uterus, rate of serious complications, and the need for additional interventions. Conservative treatment was successful in 63 patients with nontubal ectopic pregnancies. One patient had rupture of cornual pregnancy and underwent cornual resection. None of the patients in this case series required hysterectomy. This series included four patients with heterotopic pregnancies, three of whom continued intrauterine pregnancy to term gestation after conservative treatment. Seven patients experienced minimal morbidity that was treated with additional nonsurgical interventions. Conservative management and fertility preservation is feasible in most nontubal ectopic pregnancies.


An accurate, sensitive, and scalable method to identify functional sites in protein structures.
Functional sites determine the activity and interactions of proteins and as such constitute the targets of most drugs. However, the exponential growth of sequence and structure data far exceeds the ability of experimental techniques to identify their locations and key amino acids. To fill this gap we developed a computational Evolutionary Trace method that ranks the evolutionary importance of amino acids in protein sequences. Studies show that the best-ranked residues form fewer and larger structural clusters than expected by chance and overlap with functional sites, but until now the significance of this overlap has remained qualitative. Here, we use 86 diverse protein structures, including 20 determined by the structural genomics initiative, to show that this overlap is a recurrent and statistically significant feature. An automated ET correctly identifies seven of ten functional sites by the least favorable statistical measure, and nine of ten by the most favorable one. These results quantitatively demonstrate that a large fraction of functional sites in the proteome may be accurately identified from sequence and structure. This should help focus structure-function studies, rational drug design, protein engineering, and functional annotation to the relevant regions of a protein.


Transdermal Buprenorphine Relieves Neuropathic Pain: A Randomized, Double-Blind, Parallel-Group, Placebo-Controlled Trial in Diabetic Peripheral Neuropathic Pain.
To evaluate the efficacy and safety of transdermal buprenorphine in patients with diabetic peripheral neuropathic pain (DPNP). This multicenter, randomized, double-blind, placebo-controlled, parallel-group trial enrolled patients with type 1 or type 2 diabetes and stable glycemic control who had been experiencing moderate to severe DPNP for at least 6 months on maximal tolerated conventional therapy. Patients were randomly assigned to receive buprenorphine (5 μg/h) or placebo patches. The dose was titrated to effect to a maximum of 40 μg/h. Paracetamol was available as rescue analgesia. The severity of pain and other symptoms of DPNP were assessed daily in a patient diary and at clinic visits. One hundred eight-six patients were enrolled, with 93 randomized to either buprenorphine or placebo. A high proportion of patients did not complete the study (buprenorphine 37 of 93, placebo 24 of 93). The main reason for premature withdrawal in the buprenorphine group was adverse events commonly due to untreated nausea and/or vomiting. Among the per-protocol population, more patients in the buprenorphine group (86.3%) experienced a 30% reduction in average versus baseline pain at week 12 than those in the placebo group (56.6%, P < 0.001). A nonsignificant trend favored the buprenorphine group within the intention-to-treat analysis of the same end point (51.7% vs. 41.3%, P = 0.175). Transdermal buprenorphine, when tolerated, is an effective therapy for DPNP and provides another option to manage this challenging painful condition. Nausea and constipation need to be managed proactively to optimize treatment outcomes.


Profound Hyperthermia After Postpartum Rectal Misoprostol Administration.
Misoprostol, a synthetic prostaglandin E1 analog, is commonly used for treatment and prevention of postpartum hemorrhage. Known side effects include transient hyperthermia, chills, nausea, vomiting, and diarrhea. After a precipitous vaginal delivery complicated by postpartum hemorrhage (600-mL blood loss), a healthy 21-year-old multiparous patient received 800 micrograms rectal misoprostol. Within 30 minutes, she developed rigors, severe hyperthermia (41.5°C [106.7°F]), tachycardia (170s), and transient encephalopathy. Antibiotics and a cooling protocol were initiated in the intensive care unit. Her abnormal vital signs resolved 7-8 hours later. Extensive workup was negative. It is important to consider misoprostol toxicity in postpartum hyperthermia, rigors, and tachycardia. Misoprostol should be used judiciously given a lack of evidence for its effectiveness and its potential for serious side effects.