Phosphorylation of histone H3T6 by PKCbeta(I) controls demethylation at histone H3K4.
Demethylation at distinct lysine residues in histone H3 by lysine-specific demethylase 1 (LSD1) causes either gene repression or activation. As a component of co-repressor complexes, LSD1 contributes to target gene repression by removing mono- and dimethyl marks from lysine 4 of histone H3 (H3K4). In contrast, during androgen receptor (AR)-activated gene expression, LSD1 removes mono- and dimethyl marks from lysine 9 of histone H3 (H3K9). Yet, the mechanisms that control this dual specificity of demethylation are unknown. Here we show that phosphorylation of histone H3 at threonine 6 (H3T6) by protein kinase C beta I (PKCbeta(I), also known as PRKCbeta) is the key event that prevents LSD1 from demethylating H3K4 during AR-dependent gene activation. In vitro, histone H3 peptides methylated at lysine 4 and phosphorylated at threonine 6 are no longer LSD1 substrates. In vivo, PKCbeta(I) co-localizes with AR and LSD1 on target gene promoters and phosphorylates H3T6 after androgen-induced gene expression. RNA interference (RNAi)-mediated knockdown of PKCbeta(I) abrogates H3T6 phosphorylation, enhances demethylation at H3K4, and inhibits AR-dependent transcription. Activation of PKCbeta(I) requires androgen-dependent recruitment of the gatekeeper kinase protein kinase C (PKC)-related kinase 1 (PRK1). Notably, increased levels of PKCbeta(I) and phosphorylated H3T6 (H3T6ph) positively correlate with high Gleason scores of prostate carcinomas, and inhibition of PKCbeta(I) blocks AR-induced tumour cell proliferation in vitro and cancer progression of tumour xenografts in vivo. Together, our data establish that androgen-dependent kinase signalling leads to the writing of the new chromatin mark H3T6ph, which in consequence prevents removal of active methyl marks from H3K4 during AR-stimulated gene expression.


The role of hepatic transferrin receptor 2 in the regulation of iron homeostasis in the body.
Fine-tuning of body iron is required to prevent diseases such as iron-overload and anemia. The putative iron sensor, transferrin receptor 2 (TfR2), is expressed in the liver and mutations in this protein result in the iron-overload disease Type III hereditary hemochromatosis (HH). With the loss of functional TfR2, the liver produces about 2-fold less of the peptide hormone hepcidin, which is responsible for negatively regulating iron uptake from the diet. This reduction in hepcidin expression leads to the slow accumulation of iron in the liver, heart, joints, and pancreas and subsequent cirrhosis, heart disease, arthritis, and diabetes. TfR2 can bind iron-loaded transferrin (Tf) in the bloodstream, and hepatocytes treated with Tf respond with a 2-fold increase in hepcidin expression through stimulation of the bone morphogenetic protein (BMP)-signaling pathway. Loss of functional TfR2 or its binding partner, the original HH protein, results in a loss of this transferrin-sensitivity. While much is known about the trafficking and regulation of TfR2, the mechanism of its transferrin-sensitivity through the BMP-signaling pathway is still not known.


Theoretical analysis of spectrally encoded endoscopy.
Using a single optical fiber and miniature distal optics, spectrally-encoded endoscopy (SEE) has been demonstrated as a promising, three-dimensional endoscopic imaging method with a large number of resolvable points and high frame rates. We present a detailed theoretical study of the SEE prototype system and probe. Several key imaging parameters of SEE are thoroughly derived and formulated, including the three-dimensional point-spread function and field of view, as well as the system's optical aberrations and fundamental limits. We find that the point-spread function of the SEE system maintains a unique relation between its transverse and axial shapes, discuss the asymmetry of the volumetric field of view, determine that the number of lateral resolvable points is nearly twice than what was previously accepted, and derive an expression for the upper limit for the total number of resolvable points in the cross-sectional image plane.


Semisupervised multitask learning.
Context plays an important role when performing classification, and in this paper we examine context from two perspectives. First, the classification of items within a single task is placed within the context of distinct concurrent or previous classification tasks (multiple distinct data collections). This is referred to as multi-task learning (MTL), and is implemented here in a statistical manner, using a simplified form of the Dirichlet process. In addition, when performing many classification tasks one has simultaneous access to all unlabeled data that must be classified, and therefore there is an opportunity to place the classification of any one feature vector within the context of all unlabeled feature vectors; this is referred to as semi-supervised learning. In this paper we integrate MTL and semi-supervised learning into a single framework, thereby exploiting two forms of contextual information. Example results are presented on a "toy" example, to demonstrate the concept, and the algorithm is also applied to three real data sets.


Analysis of Football Injuries by Position Group in Division I College Football: A 5-Year Program Review.
The purpose of this study was to evaluate injury characteristics by position groups. Prospective, observational study. A single, major Division I collegiate football program. All players on a collegiate football program each fall regular season. Exposure to Division I collegiate football and position groups. Injury rates (IRs) per 1000 athlete exposures (AEs) and injury rate ratios (IRRs) were calculated and analyzed for all monitored injury variables, which included time in the season, body part, type of injury, game and practice injuries, mechanism of injury, and type of exposure. During the 2012 to 2016 fall regular seasons, there were 200 reported injuries sustained from 48 615 AE. The overall 5-year IR was 4.11 per 1000 AEs (3.57-4.72 95% confidence intervals). Skill players sustained the highest IR in the preseason (IR, 7.56) compared with line (IR, 4.26) and other (IR, 4.10) position groups. In addition, skill players demonstrated a significantly higher IRR compared with the line (IRR, 1.75, P < 0.05) and other (IRR, 1.85, P < 0.05) position groups. Skill players sustained most of their injuries in the preseason, whereas the linemen and other position groups suffered most of their injuries in the first half of the regular season. Skill players demonstrated a significantly higher IR in preseason, noncontact mechanism injuries, and injuries to the upper leg and thigh compared with line and other position groups. Efforts to reduce soft-tissue muscle strains in skill players targeting the preseason may provide one of the best opportunities to significantly decrease current football IRs, whereas efforts to reduce contact exposures may have the greatest effect on concussions and contact mechanism injuries for the other position group. There were no significant differences in IRs between position groups and type of exposure.


N-aralkyl derivatives of 1-aminobenzotriazole as potent isozyme-selective mechanism-based inhibitors of rabbit pulmonary cytochrome P450 in vivo.
Two N-aralkylated (N-benzyl-and N-alpha-methylbenzyl-) derivatives of 1-aminobenzotriazole, a mechanism-based inhibitor of cytochrome P450 with low isozyme selectivity, were previously shown to be potent and isozyme-selective suicide substrates for rabbit and guinea pig pulmonary P450 in vitro (Mathews and Bend, 1986; Woodcroft et al., 1990). These three compounds were compared as inhibitors in vivo after i.v. administration to rabbits treated with the cytochrome P450 inducers beta-naphthoflavone or phenobarbital. By 1 hr after administration of N-alpha-methylbenzyl-1-aminobenzotriazole (1 mumol/kg), 80% of P450 2Bs-catalyzed benzphetamine N-demethylation in lung of beta-naphthoflavone-treated rabbits was inactivated and about 35% of P450 was lost without inhibition of P450 1A1-catalyzed activity; at a dose of 10 or 100 mumol/kg, this compound totally inactivated pulmonary P450 2Bs activity while exerting minimal effects on benzphetamine N-demethylation activity (< 20% inhibition) in liver of beta-naphthoflavone-treated rabbits. N-benzyl-1-aminobenzotriazole was also an isozyme- and tissue-selective inhibitor of pulmonary P450 2Bs in vivo. Relatively high doses (100 mumol/kg) of these compounds were compared in phenobarbital-induced rabbits. Virtually all (> or = 95%) of pulmonary P450 2Bs-dependent activity was inhibited by the two N-aralkylated compounds (vs. 50% for 1-aminobenzotriazole). At this dose, about 25% of hepatic P450 was destroyed by all three compounds, whereas 1-aminobenzotriazole and its N-benzyl and N-alpha-methylbenzyl derivatives inactivated 20, 50 and 85% of hepatic P450 2Bs-selective benzphetamine N-demethylation activity, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)


Muscle synergy stability and human balance maintenance.
The signals that the central nervous system (CNS) produces and sends to the muscles to effect movement are not entirely understood. Muscle synergy theory suggests that the central nervous system produces a small number of signals that pass through a network that distributes combinations of these signals to the muscles. Though these synergies are rather stable over time, some variability is present. Here, we investigated the variability of muscle synergy and defined a synergy stability index (SSI) to quantify it. We measured the activity of muscles responsible for maintaining lateral balance in humans standing on a platform that was subjected to lateral disturbance from the platform. We then calculated muscle synergies attributed to postural reflex and automatic response by using non-negative matrix factorization (NMF). Finally, from the calculated muscle synergies, we obtained SSI. We observed a positive proportional relation between balance performance and SSI. Participants who were adept at maintaining balance were found to have invariant muscle synergies, and non-adept participants showed variable muscle synergies. These results suggest that SSI can be used to quantitatively evaluate balance maintenance ability.


Legacy and emerging flame retardants (FRs) in the freshwater ecosystem: A review.
In this review article, we have compiled and reviewed the previously published available literature on environmental distribution, behaviour, fate and regional trends of legacy and emerging flame retardants (FRs) including brominated (BFRs), organo-phosphate (OPFRs), novel brominated flame retardants (NBFRs) and dechlorane plus (DP) in the freshwater ecosystem. Transport and fate is discussed briefly with the evidences of de-bromination, sedimentation and accumulation in biota. De-bromination of BDE-209 is considered of concern because the lower brominated congeners are more toxic and mobile thus posing increased risk to the freshwater ecosystem. The available data on temporal and spatial trends as yet, is too few to show any consistent trends, enabling only general conclusions to be drawn. There is a lack of temporal studies in Asia, while, overall the trends are mixed, with both increasing and decreasing concentrations of BFRs and OPFRs. OPFRs and NBFRs have replaced classical BFRs (polybrominated diphenyl ethers (PBDEs)) in some countries but the amount of PBDEs in the environment is still considerable. Knowledge gaps and recommendations for future research are discussed emphasizing on further monitoring, advanced analytical methodologies, and risk assessment studies to completely understand the science of flame retardants in the freshwater ecosystem.


Automatic generation of personalised skeletal models of the lower limb from three-dimensional bone geometries.
The generation of personalised and patient-specific musculoskeletal models is currently a cumbersome and time-consuming task that normally requires several processing hours and trained operators. We believe that this aspect discourages the use of computational models even when appropriate data are available and personalised biomechanical analysis would be beneficial. In this paper we present a computational tool that enables the fully automatic generation of skeletal models of the lower limb from three-dimensional bone geometries, normally obtained by segmentation of medical images. This tool was evaluated against four manually created lower limb models finding remarkable agreement in the computed joint parameters, well within human operator repeatability. The coordinate systems origins were identified with maximum differences between 0.5 mm (hip joint) and 5.9 mm (subtalar joint), while the joint axes presented discrepancies between 1° (knee joint) to 11° (subtalar joint). To prove the robustness of the methodology, the models were built from four datasets including both genders, anatomies ranging from juvenile to elderly and bone geometries reconstructed from high-quality computed tomography as well as lower-quality magnetic resonance imaging scans. The entire workflow, implemented in MATLAB scripting language, executed in seconds and required no operator intervention, creating lower extremity models ready to use for kinematic and kinetic analysis or as baselines for more advanced musculoskeletal modelling approaches, of which we provide some practical examples. We auspicate that this technical advancement, together with upcoming progress in medical image segmentation techniques, will promote the use of personalised models in larger-scale studies than those hitherto undertaken.


Essential role of lung plasmacytoid dendritic cells in preventing asthmatic reactions to harmless inhaled antigen.
Tolerance is the usual outcome of inhalation of harmless antigen, yet T helper (Th) type 2 cell sensitization to inhaled allergens induced by dendritic cells (DCs) is common in atopic asthma. Here, we show that both myeloid (m) and plasmacytoid (p) DCs take up inhaled antigen in the lung and present it in an immunogenic or tolerogenic form to draining node T cells. Strikingly, depletion of pDCs during inhalation of normally inert antigen led to immunoglobulin E sensitization, airway eosinophilia, goblet cell hyperplasia, and Th2 cell cytokine production, cardinal features of asthma. Furthermore, adoptive transfer of pDCs before sensitization prevented disease in a mouse asthma model. On a functional level, pDCs did not induce T cell division but suppressed the generation of effector T cells induced by mDCs. These studies show that pDCs provide intrinsic protection against inflammatory responses to harmless antigen. Therapies exploiting pDC function might be clinically effective in preventing the development of asthma.


Understanding and meeting the needs of women in the postpartum period: the Perinatal Maternal Health Promotion Model.
A new model for the care of women in the postpartum focuses on the development of life skills that promote complete well-being. The year following childbirth is a time of significant transition for women. In addition to the physiologic changes associated with the postpartum period, a woman undergoes marked psychosocial changes as she transitions into a motherhood role, reestablishes relationships, and works to meet the physical and emotional needs of her infant and other family members. It is a time when women are vulnerable to health problems directly related to childbirth and to compromised self-care, which can manifest in the development or reestablishment of unhealthy behaviors such as smoking and a sedentary lifestyle. In addition to long-term implications for women, compromised maternal health in the postpartum period is associated with suboptimal health and developmental outcomes for infants. Maternal health experts have called for a change in how care is provided for women in the postpartum period. This article presents the rationale for a health promotion approach to meeting the needs of women in the postpartum period and introduces the Perinatal Maternal Health Promotion Model. This conceptual framework is built around a definition of maternal well-being that asserts that health goes beyond merely the absence of medical complications. In the model, the core elements of a healthy postpartum are identified and include not only physical recovery but also the ability to meet individual needs and successfully transition into motherhood. These goals can best be achieved by helping women develop or strengthen 4 key individual health-promoting skills: the ability to mobilize social support, self-efficacy, positive coping strategies, and realistic expectations. While the model focuses on the woman, the health promotion approach takes into account that maternal health in this critical period affects and is affected by her family, social network, and community. Clinical implications of the model are addressed, including specific health promotion strategies that clinicians can readily incorporate into antepartum and postpartum care.


A case of myocardial muscular bridging causing severe hypotension during exercise-electrocardiography test.
Outlining the severity of the myocardial bridge (MB) is a critical step for selecting the appropriate option among medical, surgical, or angioplasty-based treatments. Invasive treatments are usually preferred if treatment-resistant symptoms are observed or ischemia is proven by tests such as fractional flow reserve or myocardial perfusion scintigraphy (MPS). In this report, we present a patient who developed severe hypotension during treadmill exercise test, even though there were no perfusion defects during adenosine-induced MPS. This case suggests MPS with adenosine is not a good choice for evaluating ischemia in MB patients, as it may cause false negative results.


Implant supported prosthesis in a patient with progeria: case report.
Prosthodontic rehabilitation can be accomplished with fixed, overdenture, complete, or implant-retained prostheses. Dental treatment overcomes the patient's functional, psychological, esthetic and phonation problems. Remaining healthy teeth may allow the dentist to fabricate a removable partial overdenture, fixed partial prosthesis or implant - supported prosthesis. The retention of a number of abutments helps maintain a positive ridge form with greater height and volume of the alveolar bone, improving masticatory performance, as well as providing a more stable prostheses. Dental patients who have medical problems need many treatment procedures. Multidisciplinary treatment planning is invaluable for patient's dental health. Progeria is a rare genetic condition where symptoms resembling aspects of aging are manifested at an early age. characteristic clinical findings of Progeria disease include abnormalities of the skin and hair in conjunction with characteristic facial features and skeletal abnormalities. The characteristic facies show protruding ears, beaked nose, thin lips with centrofacial cyanosis, prominent eyes, frontal and parietal bossing with pseudohydrocephaly, midface hypoplasia with micrognathia and large anterior fontanel. The other reported anomalies are dystrophic nails, hypertrophic scars and hypoplastic nipples. The findings that are nearly interested in dentistry are delayed dentition, anodontia, hypodontia, or crowding of teeth. This article presents the multidisciplinary dental treatment planning includes surgical, endodontic and prosthetic treatment of a patient with a history of progeria. In this case complete-arch fixed prostheses in both maxilla and mandible, supported by a combination of implants and teeth are reported.


Development of a targeted gene vector platform based on simian adenovirus serotype 24.
Efforts to develop adenovirus vectors suitable for genetic interventions in humans have identified three major limitations of the most frequently used vector prototype, human adenovirus serotype 5 (Ad5). These limitations--widespread preexisting anti-Ad5 immunity in humans, the high rate of transduction of normal nontarget tissues, and the lack of target-specific gene delivery--justify the exploration of other Ad serotypes as vector prototypes. In this paper, we describe the development of an alternative vector platform using simian Ad serotype 24 (sAd24). We found that sAd24 virions formed unstable complexes with blood coagulation factor X and, because of that, transduced the liver and other organs at low levels when administered intravenously. The overall pattern of biodistribution of sAd24 particles was similar, however, to that of Ad5, and the intravenously injected sAd24 was cleared by Kupffer cells, leading to their depletion. We modified the virus's fiber protein to design a Her2-specific derivative of sAd24 capable of infecting target human tumor cells in vitro. In the presence of neutralizing anti-Ad5 antibodies, Her2-mediated infection with targeted sAd24 compared favorably to that with the Ad5-derived vector. When used to target Her2-expressing tumors in animals, this fiber-modified vector achieved a higher level of gene transfer to metastasis-containing murine lungs than to tumor-free lungs. In aggregate, these studies provide important insights into sAd24 biology, identify its advantages and limitations as a vector prototype, and are thus essential for further development of an sAd24-based gene delivery platform.


An Empirical Model of Slope Ratio Comparisons.
Comparing slopes is a fundamental graph reading task and the aspect ratio chosen for a plot influences how easy these comparisons are to make. According to Banking to 45°, a classic design guideline first proposed and studied by Cleveland et al., aspect ratios that center slopes around 45° minimize errors in visual judgments of slope ratios. This paper revisits this earlier work. Through exploratory pilot studies that expand Cleveland et al.'s experimental design, we develop an empirical model of slope ratio estimation that fits more extreme slope ratio judgments and two common slope ratio estimation strategies. We then run two experiments to validate our model. In the first, we show that our model fits more generally than the one proposed by Cleveland et al. and we find that, in general, slope ratio errors are not minimized around 45°. In the second experiment, we explore a novel hypothesis raised by our model: that visible baselines can substantially mitigate errors made in slope judgments. We conclude with an application of our model to aspect ratio selection.


The Alpha variant was not associated with excess nosocomial SARS-CoV-2 infection in a multi-centre UK hospital study.
Recently emerging SARS-CoV-2 variants have been associated with an increased rate of transmission within the community. We sought to determine whether this also resulted in increased transmission within hospitals. We collected viral sequences and epidemiological data of patients with community and healthcare associated SARS-CoV-2 infections, sampled from 16th November 2020 to 10th January 2021, from nine hospitals participating in the COG-UK HOCI study. Outbreaks were identified using ward information, lineage and pairwise genetic differences between viral sequences. Mixed effects logistic regression analysis of 4184 sequences showed healthcare-acquired infections were no more likely to be identified as the Alpha variant than community acquired infections. Nosocomial outbreaks were investigated based on overlapping ward stay and SARS-CoV-2 genome sequence similarity. There was no significant difference in the number of patients involved in outbreaks caused by the Alpha variant compared to outbreaks caused by other lineages. We find no evidence to support it causing more nosocomial transmission than previous lineages. This suggests that the stringent infection prevention measures already in place in UK hospitals contained the spread of the Alpha variant as effectively as other less transmissible lineages, providing reassurance of their efficacy against emerging variants of concern.


Survey of diabetes professionals regarding developmental changes in diabetes self-care.
There are no empirically obtained data defining appropriate developmental expectations for the acquisition of self-care independence by children with insulin-dependent diabetes mellitus. This study surveyed diabetes professionals about their estimates of ages at which children typically master 38 diabetes skills. The 229 survey respondents represented a broad range of professions and clinical settings and had extensive experience as diabetes professionals. Mean mastery age estimates were less than 14 yr for the 38 skills assessed. Responses to each item were variable among respondents, with a mean SD of 2.1 yr in estimated mastery ages for all items. Estimated mastery ages were below the age ranges recommended by the American Diabetes Association for 14 of 20 comparable skills. Physicians generally expected the diabetes skills to be mastered at later ages than did nurses or other health-care professionals. There were no other consistent response patterns related to respondent characteristics, i.e., years of clinical experience, employment setting, current patient load, or method of survey distribution.


S-system parameter estimation for noisy metabolic profiles using newton-flow analysis.
Biochemical systems are commonly modelled by systems of ordinary differential equations (ODEs). A particular class of such models called S-systems have recently gained popularity in biochemical system modelling. The parameters of an S-system are usually estimated from time-course profiles. However, finding these estimates is a difficult computational problem. Moreover, although several methods have been recently proposed to solve this problem for ideal profiles, relatively little progress has been reported for noisy profiles. We describe a special feature of a Newton-flow optimisation problem associated with S-system parameter estimation. This enables us to significantly reduce the search space, and also lends itself to parameter estimation for noisy data. We illustrate the applicability of our method by applying it to noisy time-course data synthetically produced from previously published 4- and 30-dimensional S-systems. In addition, we propose an extension of our method that allows the detection of network topologies for small S-systems. We introduce a new method for estimating S-system parameters from time-course profiles. We show that the performance of this method compares favorably with competing methods for ideal profiles, and that it also allows the determination of parameters for noisy profiles.


Perceptions of Health Communication, Water Treatment and Sanitation in Artibonite Department, Haiti, March-April 2012.
The international response to Haiti's ongoing cholera outbreak has been multifaceted, including health education efforts by community health workers and the distribution of free water treatment products. Artibonite Department was the first region affected by the outbreak. Numerous organizations have been involved in cholera response efforts in Haiti with many focusing on efforts to improve water, sanitation, and hygiene (WASH). Multiple types of water treatment products have been distributed, creating the potential for confusion over correct dosage and water treatment methods. We utilized qualitative methods in Artibonite to determine the population's response to WASH messages, use and acceptability of water treatment products, and water treatment and sanitation knowledge, attitudes and practices at the household level. We conducted eighteen focus group discussions (FGDs): 17 FGDs were held with community members (nine among females, eight among males); one FGD was held with community health workers. Health messages related to WASH were well-retained, with reported improvements in hand-washing. Community health workers were identified as valued sources of health information. Most participants noted a paucity of water-treatment products. Sanitation, specifically the construction of latrines, was the most commonly identified need. Lack of funds was the primary reason given for not constructing a latrine. The construction and maintenance of potable water and sanitation services is needed to ensure a sustainable change.


Relationship between serum biochemical markers of myocardial fibrosis and diastolic function at rest and with exercise in hypertrophic cardiomyopathy.
Recent studies have demonstrated the usefulness of biochemical markers of collagen turnover as markers of myocardial fibrosis in various diseases. In this study, we hypothesized that increased collagen markers in patients with hypertrophic cardiomyopathy (HCM) were correlated with diastolic function at rest and diastolic functional reserve during exercise. Thirty-six patients with HCM and 21 controls with normal left ventricular thickness were studied. Mitral septal annular velocities and mitral inflow velocities were measured at rest and during graded supine bicycle exercise (25 W, 3-minute increments) for the assessment of diastolic function at rest and during exercise. By radioimmunoassay, a byproduct of collagen III synthesis (PIIINP) and peptides resulting from collagen I synthesis (PINP) and degradation (ICTP) were measured. The patients with HCM were divided into two groups according to the median value of the PINP/ICTP ratio in the group. At rest, the mitral annular early diastolic velocity (E') was lower and the E/E' was higher in the patients with HCM with high a PINP/ICTP ratio compared with patients with HCM with a low PINP/ICTP ratio and controls (p<0.001, p=0.012). With exercise, the Doppler parameters were increased in all groups, but there was no significant difference in the change in E' and E/E' during exercise according to collagen turnover markers. A higher PINP/ICTP ratio was associated with resting diastolic dysfunction in patients with HCM; however, there was no relationship with augmented diastolic dysfunction during exercise. We suggest that the type I collagen synthesis-to-degradation ratio is a useful marker of resting diastolic function in patients with HCM.


Detection of protein orientation on the silica microsphere surface using transverse electric/transverse magnetic whispering gallery modes.
The state of adsorbed protein molecules can be examined by comparing the shifts in a narrow line resonance wavelength of transverse electric (TE) and transverse magnetic (TM) whispering gallery modes (WGM) when the molecules adsorb onto a transparent microsphere that houses WGM. In adsorption of bovine serum albumin (BSA) onto an aminopropyl-modified silica microsphere, the TM/TE shift ratio indicated highly anisotropic polarizability of BSA in the direction normal to the surface, most likely ascribed to anchoring the heart-shaped protein molecule by one of its tips. The polarization-dependent resonance shift was confirmed when the surrounding refractive index was uniformly changed by adding salt, which would simulate adsorption of large objects.


Multisensor data fusion in an integrated tracking system for endoscopic surgery.
Surgical planning and navigation systems are vital for minimally invasive endoscopic surgeries but it is challenging to track the position and orientation of intrabody surgical instruments in these procedures. In order to address this problem, we propose a tracking system including multiple-sensor integration and data fusion. The proposed tracking approach is free of the constraints of line-of-sight, less subject to environmental distortion, and with higher update rate. By incorporating electromagnetic and inertial sensors, the system yields continuous 6-DOF information. Based on a system dynamic model and estimation theories, a new multisensor fusion algorithm, cascade orientation and position-estimation algorithm, is proposed for the integrated tracking device. The experimental results show that the proposed algorithms achieve accurate orientation and position tracking with robustness.


Measurement error in epidemiologic studies of air pollution based on land-use regression models.
Land-use regression (LUR) models are increasingly used to estimate air pollution exposure in epidemiologic studies. These models use air pollution measurements taken at a small set of locations and modeling based on geographical covariates for which data are available at all study participant locations. The process of LUR model development commonly includes a variable selection procedure. When LUR model predictions are used as explanatory variables in a model for a health outcome, measurement error can lead to bias of the regression coefficients and to inflation of their variance. In previous studies dealing with spatial predictions of air pollution, bias was shown to be small while most of the effect of measurement error was on the variance. In this study, we show that in realistic cases where LUR models are applied to health data, bias in health-effect estimates can be substantial. This bias depends on the number of air pollution measurement sites, the number of available predictors for model selection, and the amount of explainable variability in the true exposure. These results should be taken into account when interpreting health effects from studies that used LUR models.


S100 family members and trypsinogens are predictors of distant metastasis and survival in early-stage non-small cell lung cancer.
Distant metastasis is the predominant cause of death in early-stage non-small cell lung cancer (NSCLC). Currently, it is impossible to predict the occurrence of metastasis at early stages and thereby separate patients who could be cured by surgical resection alone from patients who would benefit from additional chemotherapy. In this study, we applied a comparative microarray approach to identify gene expression differences between early-stage NSCLC patients whose cancer ultimately did or did not metastasize during the course of their disease. Transcriptional profiling of 82 microarrays from two patient groups revealed differential expression of several gene families including known predictors of metastasis (e.g., matrix metalloproteinases). In addition, we found S100P, S100A2, trypsinogen C (TRY6), and trypsinogen IVb (PRSS3) to be overexpressed in tumors that metastasized during the course of the disease. In a third group of 42 patients, we confirmed the induction of S100 proteins and trypsinogens in metastasizing tumors and its significant correlation with survival by real-time quantitative reverse transcription-PCR. Overexpression of S100A2, S100P, or PRSS3 in NSCLC cell cultures led to increased transendothelial migration, corroborating the role of S100A2, S100P, and PRSS3 in the metastatic process. Taken together, we provide evidence that expression of S100 proteins and trypsinogens is associated with metastasis and predicts survival in early stages of NSCLC. For the first time, this implicates a role of S100 proteins and trypsinogens in the metastatic process of early-stage NSCLC.


Allosteric vs. spontaneous exit-site (E-site) tRNA dissociation early in protein synthesis.
During protein synthesis, deacylated transfer RNAs leave the ribosome via an exit (E) site after mRNA translocation. How the ribosome regulates tRNA dissociation and whether functional linkages between the aminoacyl (A) and E sites modulate the dynamics of protein synthesis have long been debated. Using single molecule fluorescence resonance energy transfer experiments, we find that, during early cycles of protein elongation, tRNAs are often held in the E site until being allosterically released when the next aminoacyl tRNA binds to the A site. This process is regulated by the length and sequence of the nascent peptide and by the conformational state, detected by tRNA proximity, prior to translocation. In later cycles, E-site tRNA dissociates spontaneously. Our results suggest that the distribution of pretranslocation tRNA states and posttranslocation pathways are correlated within each elongation cycle via communication between distant subdomains in the ribosome, but that this correlation between elongation cycle intermediates does not persist into succeeding cycles.


Tolerance to Citrus mosaic virus in Transgenic Trifoliate Orange Lines Harboring Capsid Polyprotein Gene.
Trifoliate orange plants (Poncirus trifoliata) were transformed with a binary vector containing the capsid polyprotein (pCP) gene of Citrus mosaic virus (CiMV) via Agrobacterium tumefaciens LBA4404. Transformation was performed on the epicotyl segments obtained from a young seedling that was grown in the dark. Southern blot hybridization analysis showed that the transgene was stable in the transgenic lines after regeneration and propagation by grafting. Transgenic lines were screened for tolerance to CiMV by mechanical inoculation. Infection was monitored 30, 60, 90, and 120 days after inoculation by reverse transcription-polymerase chain reaction. The transgenic line 24 had the lowest infection rate (7.1%) at 60 days after inoculation, in contrast to that of nontransgenic plants (65.1%).The response of other lines to inoculation ranged from susceptibility to moderate tolerance.


Local macroeconomic trends and hospital admissions for child abuse, 2000-2009.
To examine the relationship between local macroeconomic indicators and physical abuse admission rates to pediatric hospitals over time. Retrospective study of children admitted to 38 hospitals in the Pediatric Hospital Information System database. Hospital data were linked to unemployment, mortgage delinquency, and foreclosure data for the associated metropolitan statistical areas. Primary outcomes were admission rates for (1) physical abuse in children <6 years old, (2) non-birth, non-motor vehicle crash-related traumatic brain injury (TBI) in infants <1 year old (which carry high risk for abuse), and (3) all-cause injuries. Poisson fixed-effects regression estimated trends in admission rates and associations between those rates and trends in unemployment, mortgage delinquency, and foreclosure. Between 2000 and 2009, rates of physical abuse and high-risk TBI admissions increased by 0.79% and 3.1% per year, respectively (P ≤ .02), whereas all-cause injury rates declined by 0.80% per year (P < .001). Abuse and high-risk TBI admission rates were associated with the current mortgage delinquency rate and with the change in delinquency and foreclosure rates from the previous year (P ≤ .03). Neither abuse nor high-risk TBI rates were associated with the current unemployment rate. The all-cause injury rate was negatively associated with unemployment, delinquency, and foreclosure rates (P ≤ .007). Multicenter hospital data show an increase in pediatric admissions for physical abuse and high-risk TBI during a time of declining all-cause injury rate. Abuse and high-risk TBI admission rates increased in relationship to local mortgage delinquency and foreclosure trends.


Bullying among trainee doctors in Southern India: a questionnaire study.
Workplace bullying is an important and serious issue in a healthcare setting because of its potential impact on the welfare of care-providers as well as the consumers. To gauge the extent of bullying among the medical community in India; as a subsidiary objective, to assess the personality trait of the bullying victims. A cross-sectional, anonymous, self-reported questionnaire survey was undertaken among a convenient sample of all the trainee doctors at a Government Medical College in Tamil Nadu, India. A questionnaire, in English with standard written explanation of bullying was used. Basic information like age, sex, job grade and the specialty in case of Postgraduates (PGs) were also collected. The results were subjected to descriptive statistical analysis and Chi-square test for comparison of frequencies. A total of 174 doctors (115 PGs and 59 junior doctors), took part in the study with a cent percent response. Nearly half of the surveyed population reported being subjected to bullying. Nearly 54 (53%) of the men and 35 (48%) of women were subjected to bullying. Significant proportions (P < 0.0001) of medical personnel and paramedical staff bullied the PGs and junior doctors, respectively. More than 85 (90%) of bullying incidents went unreported. A significant (P < 0.0001) percentage of PGs and junior doctors revealed a personality trait towards bully. Workplace bullying is common among trainee doctors and usually goes unreported.


Decision-making on an explicit risk-taking task in preadolescents with attention-deficit/hyperactivity disorder.
Inappropriate risk-taking and disadvantageous decision-making have been described as major behavioural characteristics of patients with attention-deficit/hyperactivity disorder (ADHD). However these behaviours are difficult to measure in laboratory contexts and recent studies have yielded inconsistent results which might be related to task characteristics. The present study adopted the Game of Dice Task, a test procedure in which risks are made explicit and the load on working memory is minimal. As a result, preadolescents with ADHD (N = 23) made significantly more risky choices and suffered major losses of money compared to normal controls (N = 24) but only when they played the game a second time. Differences in risk-taking correlated significantly with hyperactivity as rated by parents and with inhibitory control, but not with working memory performance. The results are discussed in the context of current theories of ADHD.


Current Trends and Results of Endovascular Treatment of Unruptured Intracranial Aneurysms at a Single Institution in the Flow-Diverter Era.
During the past several years, the number of unruptured aneurysms treated with endovascular techniques has increased. Traditionally, coil embolization was the treatment of choice for these lesions; however, recently flow diversion has become a viable, and in some cases superior, treatment option. The current single-center study presents results and trends of endovascular treatment with flow diversion and coil embolization in an unselected group of patients with unruptured intracranial aneurysms in a "real world" setting during the flow-diverter era. Three hundred ten patients with 318 treated unruptured aneurysms from June 2009 to May 2015 were included. Patient demographics, clinical characteristics, aneurysm/treatment characteristics, and outcomes were collected prospectively. We studied the following: intensive care unit admission/reasons, perioperative and mid-/long-term complications, target aneurysm rupture, retreatment/recurrence rates, and long-term neurologic outcome using the mRS. The flow-diverter group had a larger mean aneurysm size (12.3 ± 8.6 mm versus 8.7 ± 6.3 mm, P < .0001). There were no significant differences in the immediate (P = .43) and mid-/long-term complication rates (P = .54) between groups. Periprocedural neurologic morbidity and mortality rates were 2.1% and 0.5% in the coiling group and 2.5% and 1.6% in the flow-diverter group. Patients with coiling were more likely to be retreated than those with flow diversion (14.8% versus 5.7%, P = .009). Worsening of the mRS due to the target aneurysm was noted in only 3.2% of patients. The endovascular treatment of unruptured aneurysms can be performed with very low rates of neurologic complications. Both flow-diverter and coil therapy were safe and effective.


Patients with celiac disease have a lower prevalence of non-insulin-dependent diabetes mellitus and metabolic syndrome.
We investigated whether risk for non-insulin-dependent diabetes mellitus (NIDDM) and metabolic syndrome are affected by celiac disease. We examined the prevalence of NIDDM and metabolic syndrome among adults with celiac disease, compared with matched controls. We assessed medical records of 840 patients with biopsy-proven celiac disease for diagnoses of NIDDM, hypertension, or hyperlipidemia; body mass index (BMI); lipid profile; and levels of glucose or glycosylated hemoglobin, to identify those with metabolic syndrome. Patients without celiac disease were matched for age, sex, and ethnicity (n = 840 controls). The prevalence of NIDDM and metabolic syndrome in the celiac disease cohort was compared with that of the controls and subjects included in the National Health and Nutrition Examination Survey. Twenty-six patients with celiac disease (3.1%) had NIDDM compared with 81 controls (9.6%) (P < .0001). Similarly, the prevalence of metabolic syndrome was significantly lower among patients with celiac disease than controls (3.5% vs 12.7%; P < .0001). The mean BMI of patients with celiac disease was significantly lower than that of controls (24.7 vs 27.5; P < .0001). However, celiac disease was still associated with a lower risk of NIDDM, after controlling for BMI. The prevalence of NIDDM and metabolic syndrome are lower among patients with celiac disease than in matched controls and the general population. These differences are not explained by differences in BMI. Studies are needed to determine the mechanisms by which celiac disease affects the risk for NIDDM and metabolic syndrome.


Cell surface expression of heat shock protein gp96 enhances cross-presentation of cellular antigens and the generation of tumor-specific T cell memory.
Gp96 is an endoplasmic reticular heat shock protein (HSP). We have shown previously that surface expression of gp96 (96tm) on tumor cells led to the activation of dendritic cells and increased anti-tumor immunity. In this report, we have found that protective immunity elicited by 96tm+ tumor cells was tumor-specific and long-lasting. Both CD4+ and CD8+ T cell memory were elicited. By immunizing with tumor cells loaded with the chicken ovalbumin (ova) model antigen, we demonstrated that the priming of adoptively transferred ova-specific CD8+ T cells could occur across MHC haplotypes. The efficiency of this cross priming can be significantly increased when mice were immunized with whole cells that express both ova and cell surface gp96 (ova+96tm+). Mere mixture of soluble ova with 96tm-expressing tumor cells (ova-96tm+) was insufficient, arguing for further processing of ova and perhaps the participation of 96tm-ova complexes in this process. We further compared the relative efficiency of two whole cell vaccines based on the manipulation of gp96 expression in one system: 96tm+ whole cells and cells that secrete the gp96-Ig fusion protein. We found that both vaccines are effective in a prophylactic model against tumors. Our study has reinforced the notion that the manipulation of the site of expression of HSPs may be an effective approach for cancer immunotherapy.


Ratiometric Fluorescence Platform Based on Modified Silicon Quantum Dots and Its Logic Gate Performance.
A novel optical nanoprobe based on silicon quantum dots (SiQDs) has been assembled through a one-pot low-temperature (40 °C) treatment by using 3-(aminopropyl)trimethoxysilane (APTMS) and ascorbic acid (AA) as two precursors. The water-soluble SiQDs demonstrate intense green luminescence in aqueous environment and the excitation-dependent feature has been explored. Meanwhile, the incorporation of salicylaldehyde (SA) serves to suppress the emission of SiQDs effectively via nucleophilic reaction and an "on-off" change is observed. Furthermore, the addition of Zn2+ can lead to evolution of emission peaks, and the green band at 500 nm gradually shifts toward the blue side at 455 nm. The corresponding ratiometric signal changes ( I455/ I500) can accurately determine the Zn2+ concentration and the limit of detection is calculated to be 0.17 μM in the linear range between 1 and 100 μM. In this research, a molecular logic gate (AND) system has been well established by using SA and Zn2+ as two inputs. The fluorescence emission changes based on SiQDs will shed new light on the development of functional sensors at the nanoscale level.


Direct evidence of charge separation in a metal-organic framework: efficient and selective photocatalytic oxidative coupling of amines via charge and energy transfer.
The selective aerobic oxidative coupling of amines under mild conditions is an important laboratory and commercial procedure yet a great challenge. In this work, a porphyrinic metal-organic framework, PCN-222, was employed to catalyze the reaction. Upon visible light irradiation, the semiconductor-like behavior of PCN-222 initiates charge separation, evidently generating oxygen-centered active sites in Zr-oxo clusters indicated by enhanced porphyrin π-cation radical signals. The photogenerated electrons and holes further activate oxygen and amines, respectively, to give the corresponding redox products, both of which have been detected for the first time. The porphyrin motifs generate singlet oxygen based on energy transfer to further promote the reaction. As a result, PCN-222 exhibits excellent photocatalytic activity, selectivity and recyclability, far superior to its organic counterpart, for the reaction under ambient conditions via combined energy and charge transfer.


The medicalization of love.
Pharmaceuticals or other emerging technologies could be used to enhance (or diminish) feelings of lust, attraction, and attachment in adult romantic partnerships. Although such interventions could conceivably be used to promote individual (and couple) well-being, their widespread development and/or adoption might lead to the 'medicalization' of human love and heartache--for some, a source of a serious concern. In this essay, we argue that the medicalization of love need not necessarily be problematic, on balance, but could plausibly be expected to have either good or bad consequences depending upon how it unfolds. By anticipating some of the specific ways in which these technologies could yield unwanted outcomes, bioethicists and others can help to direct the course of love's medicalization--should it happen to occur--more toward the 'good' side than the 'bad.'


A novel B cell population revealed by a CD38/CD24 gating strategy: CD38(-)CD24 (-) B cells in centenarian offspring and elderly people.
The B cell arm of adaptive immunity undergoes significant modifications with age. Elderly people are characterized by impaired B cell responses reflected in a reduced ability to effectively respond against viruses and bacteria. Alterations of immunity with advancing age (immunosenescence) have been widely studied in centenarians who are considered a good example of successful aging. In recent years, attention has shifted to centenarian offspring (CO) as a model of people genetically advantaged for healthy aging and longevity. Here, we describe the preliminary characterization of a proposed new population of memory B cells, defined as CD19(+)CD38(-)CD24(-), which we find at higher frequencies in the elderly but less so in CO than healthy age-matched random controls. In addition, we found a decreased expression of RP105 (CD180), a toll-like receptor-associated molecule, on these cells. CD180 downregulation may potentially be a marker of immunosenescence. Moreover, we show that these CD19(+)CD38(-)CD24(-) B cells produce TNF and hypothesize that their observed expansion in the elderly might contribute to the increased inflammatory status sometimes designated "inflamm-aging."


Macroscopic findings in collagenous colitis: a multi-center, retrospective, observational cohort study.
Collagenous colitis (CC) is by definition a histological diagnosis. However, colonoscopy often reveals characteristic endoscopic findings. The aim of this study was to evaluate the frequency and type of endoscopic findings in patients diagnosed with CC in 4 participating centers. This was a retrospective study; the databases of 2 university hospitals in Edinburgh (Scotland) and Malmö (Sweden), and 2 district general hospitals in Tomelloso (Spain) and Gateshead (England) were interrogated for patients diagnosed with CC between May 2008 and August 2013. Endoscopy reports and images were retrieved and reviewed; data on lesions, sedation, bowel preparation and endoscopist experience were abstracted. Categorical data are reported as mean±SD. Fischer's exact, chi-square and t (unpaired) tests were used to compare datasets. A two-tailed P-value of <0.05 was considered statistically significant. 607 patients (149 male, mean age 66.9±12.25 years) were diagnosed with CC. A total of 108/607 (17.8%) patients had one or more suggestive endoscopy findings: i.e., mucosal erythema/edema, 91/607 (15%); linear colonic mucosal defects, 12/607 (2%); or mucosal scarring, 5/607 (0.82%). For colonic mucosa erythema, there was no difference in the odds of finding erythema with the use of different bowel preparation methods (P=0.997). For colonic mucosal defects there was some evidence (P=0.005) that patients colonoscoped by experienced endoscopists had 87% less odds of developing such defects. Moreover, there was evidence that analgesia reduced the odds of developing mucosal defects by 84%. A significant minority of patients with CC have endoscopic findings in colonoscopy. The description of such findings appears to be related to the endoscopist's experience.


Use of Muscle Graft Versus Fascia Flap as Interpositional Material in Prevention of Reossification in a Rat Model of Skull Bone Defect.
To investigate the utility of two different interpositional materials (muscle graft vs. fascia flap) for preventing the osseous reunion of skull bone defect including the coronal suture line in rats. A total of 32 male Sprague-Dawley rats were divided into 2 groups (n=16 for each) after the formation of bilateral coronal bone defect, based on the interpositional materials used to prevent re-ossification; the rats were divided into the muscle graft (MG) group and the fascial flap (FF) group. In each group, the other side of the coronal suture served as the control. The rats were sacrificed at postoperative 4 weeks or 8 weeks for histopathological, radiological, and microbiologic investigations. At postoperative 8 weeks, there was partial reunion in the defects with bony tissue in both the groups; no obvious differences were noted between the groups on radiological examination.The defect content involved bone and fibrous tissue in the MG group and bony bridges and loose connective tissue in the FF group. New bone formation was moderate, marked, and extreme and the reduction in defect size was marked, moderate, and extreme in the MG, FF, and control groups, respectively. Our findings revealed that neither the temporal MG nor the temporal FF were able to achieve complete prevention of re-ossification of the skull bone defects including the coronal suture line; further, neither material was superior to the other.


Deciphering the adjustment between environment and life history in annuals: lessons from a geographically-explicit approach in Arabidopsis thaliana.
The role that different life-history traits may have in the process of adaptation caused by divergent selection can be assessed by using extensive collections of geographically-explicit populations. This is because adaptive phenotypic variation shifts gradually across space as a result of the geographic patterns of variation in environmental selective pressures. Hence, large-scale experiments are needed to identify relevant adaptive life-history traits as well as their relationships with putative selective agents. We conducted a field experiment with 279 geo-referenced accessions of the annual plant Arabidopsis thaliana collected across a native region of its distribution range, the Iberian Peninsula. We quantified variation in life-history traits throughout the entire life cycle. We built a geographic information system to generate an environmental data set encompassing climate, vegetation and soil data. We analysed the spatial autocorrelation patterns of environmental variables and life-history traits, as well as the relationship between environmental and phenotypic data. Almost all environmental variables were significantly spatially autocorrelated. By contrast, only two life-history traits, seed weight and flowering time, exhibited significant spatial autocorrelation. Flowering time, and to a lower extent seed weight, were the life-history traits with the highest significant correlation coefficients with environmental factors, in particular with annual mean temperature. In general, individual fitness was higher for accessions with more vigorous seed germination, higher recruitment and later flowering times. Variation in flowering time mediated by temperature appears to be the main life-history trait by which A. thaliana adjusts its life history to the varying Iberian environmental conditions. The use of extensive geographically-explicit data sets obtained from field experiments represents a powerful approach to unravel adaptive patterns of variation. In a context of current global warming, geographically-explicit approaches, evaluating the match between organisms and the environments where they live, may contribute to better assess and predict the consequences of global warming.


Facing the threat of influenza pandemic - roles of and implications to general practitioners.
The 2009 pandemic of H1N1 influenza, compounded with seasonal influenza, posed a global challenge. Despite the announcement of post-pandemic period on 10 August 2010 by the WHO, H1N1 (2009) virus would continue to circulate as a seasonal virus for some years and national health authorities should remain vigilant due to unpredictable behaviour of the virus. Majority of the world population is living in countries with inadequate resources to purchase vaccines and stockpile antiviral drugs. Basic hygienic measures such as wearing face masks and the hygienic practice of hand washing could reduce the spread of the respiratory viruses. However, the imminent issue is translating these measures into day-to-day practice. The experience from Severe Acute Respiratory Syndrome (SARS) in Hong Kong has shown that general practitioners (GPs) were willing to discharge their duties despite risks of getting infected themselves. SARS event has highlighted the inadequate interface between primary and secondary care and valuable health care resources were thus inappropriately matched to community needs.There are various ways for GPs to contribute in combating the influenza pandemic. They are prompt in detecting and monitoring epidemics and mini-epidemics of viral illnesses in the community. They can empower and raise the health literacy of the community such as advocating personal hygiene and other precautious measures. GPs could also assist in the development of protocols for primary care management of patients with flu-like illnesses and conduct clinical audits on the standards of preventive and treatment measures. GPs with adequate liaison with public health agencies would facilitate early diagnosis of patients with influenza.In this article, we summarise the primary care actions for phases 4-6 of the pandemic. We shall discuss the novel roles of GPs as alternative source of health care for patients who would otherwise be cared for in the secondary care level. The health care system would thus remain sustainable during the public health crisis.


Pharmacotherapy of substance use disorders in the neuroscience-based nomenclature (NbN).
In the field of substance use disorders (SUDs), medications are frequently labeled according to their main symptomatic effect (e.g., "anticraving drugs") or according to imprecise and sometimes old concepts related to treatment strategies (e.g., "replacement therapies", "antabuse drugs", or "substitution treatments"). By contrast, the neuroscience-based nomenclature (NbN) offers a clearer and more consistent rationale, according to which the main element of classification is based on the pharmacological mode of action of the medication. This review aims to display the different approved treatments used in SUDs, and to discuss the pros and cons of using this new conceptual framework in the field of addiction. According to the NbN classification, medications approved in the different SUDs can be classified in the different following categories: 1) nicotinic drugs; 2) GABAergic drugs; 3) opioid drugs; and 4) others. More specifically, medications can be distinguished between whether they mimic the same pharmacological action of the "substance" whose use should be stopped or reduced, or whether they target other more general pharmacological systems, that are supposed to be common to all SUDs, as they reflect the "universal" addiction process. The NbN offers obvious advantages, compared with previous classifications. In particular, it allows to no longer mix drugs with very different pharmacological targets under the same label. The main limitation of the NbN, when applied to psychopharmacology in general, and to SUDs medications in particular, is that drugs frequently have a "dirty" action, with multiple pharmacological targets. In this respect, it may be hard to classify drugs according to the NbN classification, without making the individual profile of each medicine more complex.


Potentially modifiable deployment characteristics and new-onset alcohol abuse or dependence in the US National Guard.
There is a limited amount of data examining the relation between the onset of alcohol abuse/dependence and the experiences of soldiers prior to (pre), during (peri) and after (post) military deployment. Some deployment characteristics, e.g., military unit cohesion, are potentially modifiable in the context of reducing alcohol abuse/dependence peri-/post deployment. We investigated the associations between potentially modifiable deployment characteristics and peri-/post (incident) alcohol abuse/dependence among deployed Ohio Army National Guard (OHARNG) soldiers. Using a sample of OHARNG (June, 2008 to February, 2009), eligible participants were ever been deployed and did not report alcohol abuse/dependence prior to deployment (final sample size=963). Interviews assessed soldiers' alcohol abuse/dependence, depression, PTSD, deployment related factors (e.g., exposure to warzone stressors) and three deployment characteristics (pre-deployment preparedness, unit support during deployment, and post-deployment social support). Associations between the three deployment characteristics and incident alcohol abuse/dependence (defined as abuse or dependence at any point during or after deployment) were estimated using logistic regression. Only pre-deployment preparedness was associated with incident alcohol abuse/dependence (a non-linear inverted-u shaped relation) when controlling for demographics, deployment related factors (e.g., exposure to warzone stressors), and the presence of psychopathology that exhibited peri-/post-deployment. We present these results graphically, plotting incident alcohol abuse/dependence over the levels of pre-deployment preparedness. The association between pre-deployment preparedness and alcohol abuse/dependence may be characterized as an inverted-U shaped function. Suggestions for how and whether to modify pre-deployment preparedness in an effort to reduce peri-/post-deployment alcohol abuse or dependence should await further research.


Transsphenoidal extension of heterotopic glioneuronal tissue: pathoanatomic considerations in symptomatic neonates.
In this clinical investigation, we aimed (1) to re-evaluate the nature of glioneuronal tissue with transsphenoidal extension and how it fits into the nomenclature of midline malformations and mass lesions; (2) to find out if our imaging findings support current pathoanatomic concepts of clefts and canals in the sphenoid body of newborns. In two neonates with respiratory distress due to nasopharyngeal masses, 3T MRI was performed, and CT in one of them. Imaging features were analyzed in consensus by two pediatric neuroradiologists with histological reports being available. An interdisciplinary panel compared the findings to those of case publications and differential entities from our institutional case collection. Referring to our rare case of transsphenoidal cerebral heterotopia and unique case of hypothalamic hamartoma with transsphenoidal herniation, glioneuronal heterotopia may definitely extend through the sphenoid bone. Consequently, there is reason for brain heterotopias to be labeled as such also in case of an intracranial component. Connection between heterotopic glioneuronal tissue in the nasopharynx and a hypothalamic hamartoma may go along with indistinct margins to normal brain. Neither extension through a transsphenoidal cleft nor association with a cleft palate are specific for cerebral heterotopia. Our findings support the hypothesis that transsphenoidal cerebral heterotopias do not or at least not invariably follow the route of Rathke's pouch, known as the craniopharyngeal canal. Transsphenoidal glioneuronal heterotopia should be the top differential diagnosis in MR imaging if a non-enhancing nasopharyngeal mass of an infant extends through a craniopharyngeal cleft within the intersphenoid synchondrosis.


Autoimmunity in membranous nephropathy targets aldose reductase and SOD2.
Glomerular targets of autoimmunity in human membranous nephropathy are poorly understood. Here, we used a combined proteomic approach to identify specific antibodies against podocyte proteins in both serum and glomeruli of patients with membranous nephropathy (MN). We detected specific anti-aldose reductase (AR) and anti-manganese superoxide dismutase (SOD2) IgG(4) in sera of patients with MN. We also eluted high titers of anti-AR and anti-SOD2 IgG(4) from microdissected glomeruli of three biopsies of MN kidneys but not from biopsies of other glomerulonephritides characterized by IgG deposition (five lupus nephritis and two membranoproliferative glomerulonephritis). We identified both antigens in MN biopsies but not in other renal pathologies or normal kidney. Confocal and immunoelectron microscopy (IEM) showed co-localization of anti-AR and anti-SOD2 with IgG(4) and C5b-9 in electron-dense podocyte immune deposits. Preliminary in vitro experiments showed an increase of SOD2 expression on podocyte plasma membrane after treatment with hydrogen peroxide. In conclusion, our data support AR and SOD2 as renal antigens of human MN and suggest that oxidative stress may drive glomerular SOD2 expression.


The role of p53 in vivo during skeletal muscle post-natal development and regeneration: studies in p53 knockout mice.
The tumour suppressor gene p53 is recognised as a central regulator of the cell cycle and apoptosis. Post-natally, p53 mutations are associated with many cancers and mice lacking p53 are prone to spontaneous tumour formation. The present study examines skeletal muscle formation in post-natal mice lacking p53 using two different models of skeletal muscle regeneration. The level of endogenous myogenic cell proliferation in mature skeletal muscle was examined and the time course of muscle regeneration after whole muscle transplantation or crush injury were compared in p53 (-/-) and control C57Bl/6J adult mice, using desmin and proliferating cell nuclear antigen (PCNA) immunohistochemistry and histological analysis. The pattern of inflammation, myoblast proliferation and myotube formation in regenerating p53 (-/-) skeletal muscles appears normal and similar to those in control C57Bl/6J muscle. These data indicate that p53 is not required for the regulation of myoblast proliferation, differentiation and myotube formation in vivo during myogenesis of adult skeletal muscle.


The dynamics between limited-term and lifelong coinfecting bacterial parasites in wild rodent hosts.
Interactions between coinfecting parasites may take various forms, either direct or indirect, facilitative or competitive, and may be mediated by either bottom-up or top-down mechanisms. Although each form of interaction leads to different evolutionary and ecological outcomes, it is challenging to tease them apart throughout the infection period. To establish the first step towards a mechanistic understanding of the interactions between coinfecting limited-term bacterial parasites and lifelong bacterial parasites, we studied the coinfection of Bartonella sp. (limited-term) and Mycoplasma sp. (lifelong), which commonly co-occur in wild rodents. We infected Bartonella- and Mycoplasma-free rodents with each species, and simultaneously with both, and quantified the infection dynamics and host responses. Bartonella benefited from the interaction; its infection load decreased more slowly in coinfected rodents than in rodents infected with Bartonella alone. There were no indications for bottom-up effects, but coinfected rodents experienced various changes, depending on the infection stage, in their body mass, stress levels and activity pattern, which may further affect bacterial replication and transmission. Interestingly, the infection dynamics and changes in the average coinfected rodent traits were more similar to the chronic effects of Mycoplasma infection, whereas coinfection uniquely impaired the host's physiological and behavioral stability. These results suggest that parasites with distinct life history strategies may interact, and their interaction may be asymmetric, non-additive, multifaceted and dynamic through time. Because multiple, sometimes contrasting, forms of interactions are simultaneously at play and their relative importance alternates throughout the course of infection, the overall outcome may change under different ecological conditions.


Substance Use Screening, Brief Intervention, and Referral to Treatment.
The enormous public health impact of adolescent substance use and its preventable morbidity and mortality highlight the need for the health care sector, including pediatricians and the medical home, to increase its capacity regarding adolescent substance use screening, brief intervention, and referral to treatment (SBIRT). The American Academy of Pediatrics first published a policy statement on SBIRT and adolescents in 2011 to introduce SBIRT concepts and terminology and to offer clinical guidance about available substance use screening tools and intervention procedures. This clinical report provides a simplified adolescent SBIRT clinical approach that, in combination with the accompanying updated policy statement, guides pediatricians in implementing substance use prevention, detection, assessment, and intervention practices across the varied clinical settings in which adolescents receive health care.


Pseudo solid-appearing pancreatic serous microcystic adenomas: Histologic diagnosis with the EUS core biopsy fork-tip needle.
Despite rarely, serous cystic adenoma (SCA) can assume a pseudo-solid aspect mimicking other pancreatic neoplasm as neuroendocrine tumor. EUS-FNA cytology has low diagnostic accuracy due to the scant cellularity of the collected samples. Histological diagnosis is usually made after resection. Recently, end-cutting needles for EUS-fine-needle biopsy (EUS-FNB), which obtain tissue cores by penetrating the lesions, have been developed. We aimed to assess the capability of EUS-FNB with SharkCore™ needles in the preoperative diagnosis of serous cystic adenoma pseudo-solid-appearing on imaging (Sa-SCA). Between January 2016 and January 2018, data from consecutive adult patients, who were referred for EUS-FNB of a solid pancreatic lesion and were diagnosed with having SCA, were retrieved from a single-center institutional database. Two patients were excluded because of microcystic aspect at EUS. Histological diagnosis of SCA was made by EUS-FNB in the remaining 7 patients (5 females; mean age of 62.5 years). Lesions (mean size of 19.8 mm) were hypervascular on cross-sectional imaging, slightly hyperdense magnetic resonance imaging with T2-weighted images can, and negative at 68Ga-somatostatin receptor positron emission tomography and 18fluoro-deoxyglucose positron emission tomography. EUS-FNB samples were judged adequate for a definitive diagnosis in all cases, achieving specimens suitable for histological evaluation and several ancillary stains. Histochemical positivity for periodic acid-Schiff (PAS) and PAS with diastase digestion was observed in 7/7 cases. Immunohistochemical positivity for α-inhibin (7/7), GLUT1 (6/6), MUC6 (5/5), and negativity for synaptophysin (7/7) and chromogranin A (2/2) favored SCA diagnosis. In the case of preoperative workup suspected for Sa-SCA, a "forward acquiring" needle could improve the rate of preoperative histological diagnosis.


Solution structures of potato virus X and narcissus mosaic virus from Raman optical activity.
Potato virus X (PVX) and narcissus mosaic virus (NMV) were studied using vibrational Raman optical activity (ROA) in order to obtain new information on the structures of their coat protein subunits. The ROA spectra of the two intact virions are very similar to each other and similar to that of tobacco mosaic virus (TMV) studied previously, being dominated by signals characteristic of proteins with helix bundle folds. In particular, PVX and NMV show strong positive ROA bands at approximately 1340 cm(-1) assigned to hydrated alpha-helix and perhaps originating in surface exposed helical residues, together with less strong positive ROA intensity in the range approximately 1297-1312 cm(-1) assigned to alpha-helix in a more hydrophobic environment and perhaps originating in residues at helix-helix interfaces. The positive approximately 1340 cm(-1) ROA band of TMV is less intense than those of PVX and NMV, suggesting that TMV contains less hydrated alpha-helix. Small differences in other spectral regions reflect differences in some loop, turn and side-chain compositions and conformations among the three viruses. A pattern recognition program based on principal component analysis of ROA spectra indicates that the coat protein subunit folds of PVX and NMV may be very similar to each other and similar to that of TMV. These results suggest that PVX and NMV may have coat protein subunit structures based on folds similar to the TMV helix bundle and hence that the helical architecture of the PVX and NMV particles may be similar to that of TMV but with different structural parameters.


A prospective study investigating the impact of school belonging factors on negative affect in adolescents.
School belonging, measured as a unidimensional construct, is an important predictor of negative affective problems in adolescents, including depression and anxiety symptoms. A recent study found that one such measure, the Psychological Sense of School Membership scale, actually comprises three factors: Caring Relations, Acceptance, and Rejection. We explored the relations of these factors with negative affect in a longitudinal study of 504 Australian Grade 7 and 8 students. Each school belonging factor contributed to the prediction of negative affect in cross-sectional analyses. Scores on the Acceptance factor predicted subsequent negative affect for boys and girls, even controlling for prior negative affect. For girls, the Rejection factor was also significant in the prospective analysis. These findings have implications for the design of interventions and are further confirmation that school belonging should be considered a multidimensional construct.


Educational impact of paediatric palliative simulation study days.
The use of simulation-based medical/nursing teaching is increasingly widespread. Simulation-based teaching offers an immersive learning experience where professionals can practice communication and practical skills in a safe, authentic environment. We designed a paediatric palliative simulation study day primarily aimed at nursing staff who manage these patients in the community/hospice. We believe this is the first of its kind in the UK. To establish whether attendance at a paediatric palliative simulation study day improved confidence and knowledge in management of common and/or difficult situations in palliative care. Health professionals working at local paediatric hospices or in associated specialties to palliative care were invited to attend the free 1-day course. 5 scenarios were developed by experienced health professionals working in paediatric palliative care. On the day, participants were asked to complete a questionnaire to check basic demographic data, confidence levels and knowledge (50 true/false questions). Following participation/observation of 5 scenarios, they again completed the same questionnaire regarding confidence levels and knowledge. Results were analysed with Excel and XLStat using basic demographic data and Wilcoxon signed rank two-tailed test. 57 healthcare workers participated in 5 study days. 81% (n=47) professionals described themselves as working primarily in palliative care. Only 35% (n=20) had previously experienced simulation. Based on confidence questions, attendees felt more confident in managing specific palliative scenarios (p<0.0001). Based on true/false questions prestudy and poststudy day, 86% (n=49) of participants improved their knowledge. The median improvement score for the cohort was 3 (p<0.0001). The study demonstrated a significant improvement in confidence and knowledge following the simulation course. This supports further time/financial investment in developing this type of study day. Simulation is a useful teaching adjunct in paediatric palliative care. The course also provides a valuable opportunity for professionals to network and discuss/share experiences.


Advantages and Limitations of Wearable Activity Trackers: Considerations for Patients and Clinicians.
Exercise, light physical activity, and decreased sedentary time all have been associated with health benefits following cancer diagnoses. Commercially available wearable activity trackers may help patients monitor and self-manage their behaviors to achieve these benefits. This article highlights some advantages and limitations clinicians should be aware of when discussing the use of activity trackers with cancer survivors. Limited research has assessed the accuracy of commercially available activity trackers compared to research-grade devices. Because most devices use confidential, proprietary algorithms to convert accelerometry data to meaningful output like total steps, assessing whether these algorithms account for differences in gait abnormalities, functional limitations, and different body morphologies can be difficult. Quantification of sedentary behaviors and light physical activities present additional challenges. The global market for activity trackers is growing, which presents clinicians with a tremendous opportunity to incorporate these devices into clinical practice as tools to promote activity. This article highlights important considerations about tracker accuracy and usage by cancer survivors.


Censored correlated cytokine concentrations: multivariate Tobit regression using clustered variance estimation.
Interest in cytokines as markers for the function of the immune system is increasing. Methods quantifying cytokine concentrations are often subject to detection limits, which lead to non-detectable observations and censored distributions. When distributions are skewed, geometric mean ratios (GMRs) can be used to describe the relative concentration between two cytokines, and the GMR ratio (GMRR) can be used to compare two groups. The problem is how to estimate GMRRs from censored distributions.We evaluated methods, including simple deletion and substitution, in simulated and real data. One method applies Tobit directly to the censored difference between the two cytokine log-concentrations (Diff). However, censoring is correlated to the outcome and is therefore not independent. The correlation increases as the correlation between the two log-concentrations decreases. We propose a Tobit stacking method that uses clustered variance-covariance estimation allowing homogeneous (Stackc) or inhomogeneous (Stackh) variances. We compare it with direct estimation of the bivariate Tobit likelihood function (Bitobit) and multiple imputation. We assess sensitivity to inhomogeneity and non-normality. Simulations show that deletion and substitution are empirically biased and that Diff has an empirical bias, which increases as the correlation between the log-concentrations decreases. Estimates from multiple imputation, Stackh and Bitobit are almost identical. The estimates exhibit small empirical bias for both homogeneous and inhomogeneous normal distributions. For skewed mixture and heavy-tailed distributions, they perform reasonably well if censoring is less than 30%. We recommend these methods to estimate GMRRs. At least one of the methods is available in Stata, R or SAS.


Safety, Feasibility of Intravenous and Intrathecal Injection of Autologous Bone Marrow Derived Mesenchymal Stromal Cells in Patients with Amyotrophic Lateral Sclerosis: An Open Label Phase I Clinical Trial.
Amyotrophic lateral sclerosis (ALS) is the most severe disorder within the spectrum of motor neuron diseases (MND) that has no effective treatment and a progressively fatal outcome. We have conducted two clinical trials to assess the safety and feasibility of intravenous (IV) and intrathecal (IT) injections of bone marrow derived mesenchymal stromal cells (BM-MSCs) in patients with ALS. This is an interventional/experimental study. We enrolled 14 patients that met the following inclusion criteria: definitive diagnosis of sporadic ALS, ALS Functional Rating Scale (ALS-FRS) ≥24, and ≥40% predicted forced vital capacity (FVC). All patients underwent bone marrow (BM) aspiration to obtain an adequate sample for cell isolation and culture. Patients in group 1 (n=6) received an IV and patients in group 2 (n=8) received an IT injection of the cell suspension. All patients in both groups were followed at 24 hours and 2, 4, 6, and 12 months after the injection with ALS-FRS, FVC, laboratory tests, check list of side effects and brain/spinal cord magnetic resonance imaging (MRI). In each group, one patient was lost to follow up one month after cell injection and one patient from IV group died due to severe respiratory insufficiency and infection. During the follow up there were no reports of adverse events in terms of clinical and laboratory assessments. In MRI, there was not any new abnormal finding. The ALS-FRS score and FVC percentage significantly reduced in all patients from both groups. This study has shown that IV and IT transplantation of BM-derived stromal cells is safe and feasible (Registration numbers: NCT01759797 and NCT01771640).


Clinical review: intensive care unit acquired weakness.
A substantial number of patients admitted to the ICU because of an acute illness, complicated surgery, severe trauma, or burn injury will develop a de novo form of muscle weakness during the ICU stay that is referred to as "intensive care unit acquired weakness" (ICUAW). This ICUAW evoked by critical illness can be due to axonal neuropathy, primary myopathy, or both. Underlying pathophysiological mechanisms comprise microvascular, electrical, metabolic, and bioenergetic alterations, interacting in a complex way and culminating in loss of muscle strength and/or muscle atrophy. ICUAW is typically symmetrical and affects predominantly proximal limb muscles and respiratory muscles, whereas facial and ocular muscles are often spared. The main risk factors for ICUAW include high severity of illness upon admission, sepsis, multiple organ failure, prolonged immobilization, and hyperglycemia, and also older patients have a higher risk. The role of corticosteroids and neuromuscular blocking agents remains unclear. ICUAW is diagnosed in awake and cooperative patients by bedside manual testing of muscle strength and the severity is scored by the Medical Research Council sum score. In cases of atypical clinical presentation or evolution, additional electrophysiological testing may be required for differential diagnosis. The cornerstones of prevention are aggressive treatment of sepsis, early mobilization, preventing hyperglycemia with insulin, and avoiding the use parenteral nutrition during the first week of critical illness. Weak patients clearly have worse acute outcomes and consume more healthcare resources. Recovery usually occurs within weeks or months, although it may be incomplete with weakness persisting up to 2 years after ICU discharge. Prognosis appears compromised when the cause of ICUAW involves critical illness polyneuropathy, whereas isolated critical illness myopathy may have a better prognosis. In addition, ICUAW has shown to contribute to the risk of 1-year mortality. Future research should focus on new preventive and/or therapeutic strategies for this detrimental complication of critical illness and on clarifying how ICUAW contributes to poor longer-term prognosis.


Alterations of the Innate Immune System in Susceptibility and Resilience After Social Defeat Stress.
Dysregulation of innate immune responses has frequently been reported in stress-associated psychiatric disorders such as major depression. In mice, enhanced circulating cytokine levels as well as altered innate immune cell numbers have been found after stress exposure. In addition, stress-induced recruitment of peripheral monocytes to the brain has been shown to promote anxiety-like behavior. However, it is yet unclear whether specific differences in the innate immune system are associated with stress susceptibility or resilience in mice. Utilizing chronic social defeat, a model of depression and stress vulnerability, we characterized peripheral and brain-invading myeloid cells in stress-susceptible and resilient animals. In all defeated animals, we found reduced percentages of CD11c+ dendritic cells (DCs) by flow cytometry in the spleen when compared to non-defeated controls. Exclusively in susceptible mice conventional DCs of the spleen showed up-regulated expression of MHC class II and co-stimulatory CD80 molecules pointing toward an enhanced maturation phenotype of these cells. Susceptible, but not resilient animals further exhibited an increase in inflammatory Ly6Chi monocytes and higher numbers of spleen-derived CD11b+ cells that produced the proinflammatory cytokine tumor necrosis factor (TNF) upon lipopolysaccharide (LPS) stimulation. Increased percentages of peripheral CD45hi CD11b+ cells immigrated into the brain of defeated mice, regardless of resilience or susceptibility. However, cellular infiltrates in the brain of susceptible mice contained higher percentages of CC chemokine receptor 2 (CCR2+) Ly6Chi monocytes representing an inflammatory phenotype. Thus, we defined specific stress-related immune signatures involving conventional DCs and inflammatory Ly6Chi monocytes in susceptible and resilient mice. Together, our findings suggest an impact of the innate immune system in vulnerability to stress-related disorders such as major depression.


Aberrant natural killer (NK) cell activation and dysfunction among ART-treated HIV-infected adults in an African cohort.
We examined NK cell phenotypes and functions after seven years of ART and undetectable viral loads (<50 copies/ml) with restored CD4 T-cell counts (≥500 cells/μl) and age-matched healthy-HIV-uninfected individuals from the same community. Using flow-cytometry, NK cell phenotypes were described using lineage markers (CD56+/-CD16+/-). NK cell activation was determined by expression of activation receptors (NKG2D, NKp44 and NKp46) and activation marker CD69. NK cell function was determined by CD107a, granzyme-b, and IFN-gamma production. CD56 dim and CD56 bright NK cells were lower among ART-treated-HIV-infected than among age-matched-HIV-negative individuals; p = 0.0016 and p = 0.05 respectively. Production of CD107a (P = 0.004) and Granzyme-B (P = 0.005) was lower among ART-treated-HIV-infected relative to the healthy-HIV-uninfected individuals. NKG2D and NKp46 were lower, while CD69 expression was higher among ART-treated-HIV-infected than healthy-HIV-uninfected individuals. NK cell activation and dysfunction persisted despite seven years of suppressive ART with "normalization" of peripheral CD4 counts.


Are linear measurements and computerized volumetric ratios determined from axial MRI useful for diagnosing hydrocephalus in children with tuberculous meningitis?
This study aimed to evaluate linear measurements and computerized volumetric ratios on axial magnetic resonance imaging (MRI) scans against the diagnosis of hydrocephalus in children with tuberculous meningitis (TBM). MRI scans and clinical notes of children with culture positive TBM were reviewed. Patients with surgical drainage of ventricles were considered positive for hydrocephalus. Alternatively, predefined radiological criteria of hydrocephalus in combination with any clinical criteria were considered positive for hydrocephalus. Axial T2-weighted MRI scans were used for measurement by a radiologist. Linear measurements included the Evans index, frontal-occipital horn ratio, and frontal-occipital horn width ratio. Computer-assisted segmentation of the MRI volume was performed on a slice-by-slice basis using the number of pixels comprising each region to calculate the ratios: ventricular volume: brain volume and cerebrospinal fluid (CSF)/(brain + CSF) for all slices and for a single slice at the level of the lateral ventricles. Twenty-two children (mean age 3.7 years) comprised ten patients with a 'final' diagnosis of hydrocephalus (six communicating, four non-communicating). None of the linear measurements showed a statistical correlation with the 'final' diagnosis of hydrocephalus. The frontal-occipital horn width ratio (FOHWR) (p = 0.09) was the closest to demonstrate statistical significance. The highest sensitivity was attained with FOR (90%) followed by FOHWR (85%). The highest specificity was reached with FOHWR (70%). Volumetric ratios were inferior to linear measures. Linear measures of hydrocephalus in TBM were more reliable than volumetric ratios. Hydrocephalus can be quantified most reliably using the FOHWR. This is useful for serial follow-up and for research of TBM.


Identifying postpartum intervention approaches to reduce cardiometabolic risk among American Indian women with prior gestational diabetes, Oklahoma, 2012-2013.
Innovative approaches are needed to reduce cardiometabolic risk among American Indian women with a history of gestational diabetes. We assessed beliefs of Oklahoma American Indian women about preventing type 2 diabetes and cardiovascular disease after having gestational diabetes. We also assessed barriers and facilitators to healthy lifestyle changes postpartum and intervention approaches that facilitate participation in a postpartum lifestyle program. In partnership with a tribal health system, we conducted a mixed-method study with American Indian women aged 19 to 45 years who had prior gestational diabetes, using questionnaires, focus groups, and individual interviews. Questionnaires were used to identify women's cardiometabolic risk perceptions and feasibility and acceptability of Internet or mobile phone technology for delivery of a postpartum lifestyle modification program. Focus groups and individual interviews were conducted to identify key perspectives and preferences related to a potential program. Participants were 26 women, all of whom completed surveys; 11 women participated in focus group sessions, and 15 participated in individual interviews. Most women believed they would inevitably develop diabetes, cardiovascular disease, or both; however, they were optimistic that they could delay onset with lifestyle change. Most women expressed enthusiasm for a family focused, technology-based intervention that emphasizes the importance of delaying disease onset, provides motivation, and promotes accountability while accommodating women's competing priorities. Our findings suggest that an intervention that uses the Internet, text messaging, or both and that emphasizes the benefits of delaying disease onset should be tested as a novel, culturally relevant approach to reducing rates of diabetes and cardiovascular disease in this high-risk population.


The Insula Mediates Access to Awareness of Visual Stimuli Presented Synchronously to the Heartbeat.
The processing of interoceptive signals in the insular cortex is thought to underlie self-awareness. However, the influence of interoception on visual awareness and the role of the insular cortex in this process remain unclear. Here, we show in a series of experiments that the relative timing of visual stimuli with respect to the heartbeat modulates visual awareness. We used two masking techniques and show that conscious access for visual stimuli synchronous to participants' heartbeat is suppressed compared with the same stimuli presented asynchronously to their heartbeat. Two independent brain imaging experiments using high-resolution fMRI revealed that the insular cortex was sensitive to both visible and invisible cardio-visual stimulation, showing reduced activation for visual stimuli presented synchronously to the heartbeat. Our results show that interoceptive insular processing affects visual awareness, demonstrating the role of the insula in integrating interoceptive and exteroceptive signals and in the processing of conscious signals beyond self-awareness. There is growing evidence that interoceptive signals conveying information regarding the internal state of the body influence perception and self-awareness. The insular cortex, which receives sensory inputs from both interoceptive and exteroceptive sources, is thought to integrate these multimodal signals. This study shows that cardiac interoceptive signals modulate awareness for visual stimuli such that visual stimuli occurring at the cardiac frequency take longer to access visual awareness and are more difficult to discriminate. Two fMRI experiments show that the insular region is sensitive to this cardio-visual synchrony even when the visual stimuli are rendered invisible through interocular masking. The results indicate a perceptual and neural suppression for visual events coinciding with cardiac interoceptive signals.


Identification of cardioprotective drugs by medium-scale in vivo pharmacological screening on a Drosophila cardiac model of Friedreich's ataxia.
Friedreich's ataxia (FA) is caused by reduced levels of frataxin, a highly conserved mitochondrial protein. There is currently no effective treatment for this disease, which is characterized by progressive neurodegeneration and cardiomyopathy, the latter being the most common cause of death in patients. We previously developed a Drosophila melanogaster cardiac model of FA, in which the fly frataxin is inactivated specifically in the heart, leading to heart dilatation and impaired systolic function. Methylene Blue (MB) was highly efficient to prevent these cardiac dysfunctions. Here, we used this model to screen in vivo the Prestwick Chemical Library, comprising 1280 compounds. Eleven drugs significantly reduced the cardiac dilatation, some of which may possibly lead to therapeutic applications in the future. The one with the strongest protective effects was paclitaxel, a microtubule-stabilizing drug. In parallel, we characterized the histological defects induced by frataxin deficiency in cardiomyocytes and observed strong sarcomere alterations with loss of striation of actin fibers, along with full disruption of the microtubule network. Paclitaxel and MB both improved these structural defects. Therefore, we propose that frataxin inactivation induces cardiac dysfunction through impaired sarcomere assembly or renewal due to microtubule destabilization, without excluding additional mechanisms. This study is the first drug screening of this extent performed in vivo on a Drosophila model of cardiac disease. Thus, it also brings the proof of concept that cardiac functional imaging in adult Drosophila flies is usable for medium-scale in vivo pharmacological screening, with potent identification of cardioprotective drugs in various contexts of cardiac diseases.


Distribution and Stimulus Secretion Coupling of Enteroendocrine Cells along the Intestinal Tract.
The enteroendocrine system of the gut acts both locally and peripherally, regulating gastrointestinal function as well as metabolism, energy expenditure, and central appetite control through the release of a variety of hormones. The chemosensing ability of enteroendocrine cells is integral to their role in eliciting physiological changes in response to fluctuations in the composition of the intestinal lumen. Regulation of enteroendocrine cell activity is complex, and requires that these cells can integrate signals deriving from dietary sources as well as the nervous and endocrine systems. Here, we provide an overview of enteroendocrine cell form and function, with a focus on new insights into their distribution throughout the intestine and the stimulus secretion coupling mechanisms underlying the activity of these important members of the gut-brain axis. © 2018 American Physiological Society. Compr Physiol 8:1603-1638, 2018.


A protein kinase A-independent pathway controlling aquaporin 2 trafficking as a possible cause for the syndrome of inappropriate antidiuresis associated with polycystic kidney disease 1 haploinsufficiency.
Renal water reabsorption is controlled by arginine vasopressin (AVP), which binds to V2 receptors, resulting in protein kinase A (PKA) activation, phosphorylation of aquaporin 2 (AQP2) at serine 256, and translocation of AQP2 to the plasma membrane. However, AVP also causes dephosphorylation of AQP2 at S261. Recent studies showed that cyclin-dependent kinases (cdks) can phosphorylate AQP2 peptides at S261 in vitro. We investigated the possible role of cdks in the phosphorylation of AQP2 and identified a new PKA-independent pathway regulating AQP2 trafficking. In ex vivo kidney slices and MDCK-AQP2 cells, R-roscovitine, a specific inhibitor of cdks, increased pS256 levels and decreased pS261 levels. The changes in AQP2 phosphorylation status were paralleled by increases in cell surface expression of AQP2 and osmotic water permeability in the absence of forskolin stimulation. R-Roscovitine did not alter cAMP-dependent PKA activity but specifically reduced protein phosphatase 2A (PP2A) expression and activity in MDCK cells. Notably, we found reduced PP2A expression and activity and reduced pS261 levels in Pkd1(+/-) mice displaying a syndrome of inappropriate antidiuresis with high levels of pS256, despite unchanged AVP and cAMP. Similar to previous findings in Pkd1(+/-) mice, R-roscovitine treatment caused a significant decrease in intracellular calcium in MDCK cells. Our data indicate that reduced activity of PP2A, secondary to reduced intracellular Ca(2+) levels, promotes AQP2 trafficking independent of the AVP-PKA axis. This pathway may be relevant for explaining pathologic states characterized by inappropriate AVP secretion and positive water balance.