Optical Coherence Tomography Angiography Reveals Spatial Bias of Macular Capillary Dropout in Diabetic Retinopathy.
Our purpose is to evaluate the spatial bias of macular capillary dropout accompanying diabetic retinopathy (DR) using optical coherence tomography angiography (OCTA). This study included 47 patients with diabetes and 29 healthy individuals who underwent OCTA. Retinal capillary flow density (FD) of 2.6 × 2.6 or 5.2 × 5.2 mm foveal area as well as the four divided areas (superior, inferior, temporal, nasal) without a foveal avascular zone (FAZ) at the superficial capillary plexus and deep capillary plexus (DCP) were measured respectively using ImageJ and NI Vision. Spatial biases of FD (orientation bias ratio and hierarchical bias ratio) and the correlation between FAZ and FD were examined. OCTA showed focal capillary dropout in DR patients. The orientation bias of FD was significantly higher in NPDR compared to NDR in the DCP (P = 0.03). The hierarchical bias of FD was significantly shifted to a DCP dominance with progression of DR (P < 0.01). In addition, the FD and FAZ area were significantly inversely correlated in both plexus in DR patients but not in healthy subjects (P < 0.01). Area-divided OCTA quantification shows the appearance of spatial biases of macular capillary dropout with the onset of DR, suggesting that DR-related macular capillary dropout occurs locally and randomly. Future studies are necessary to determine the clinical relevance of the spatial pattern of capillary dropout in DR.


Integrating Bayesian Belief Networks in a toolbox for decision support on plastic clean-up technologies in rivers and estuaries.
Current mitigation strategies to offset marine plastic pollution, a global concern, typically rely on preventing floating debris from reaching coastal ecosystems. Specifically, clean-up technologies are designed to collect plastics by removing debris from the aquatic environment such as rivers and estuaries. However, to date, there is little published data on their potential impact on riverine and estuarine organisms and ecosystems. Multiple parameters might play a role in the chances of biota and organic debris being unintentionally caught within a mechanical clean-up system, but their exact contribution to a potential impact is unknown. Here, we identified four clusters of parameters that can potentially determine the bycatch: (i) the environmental conditions in which the clean-up system is deployed, (ii) the traits of the biota the system interacts with, (iii) the traits of plastic items present in the system, and, (iv) the design and operation of the clean-up mechanism itself. To efficiently quantify and assess the influence of each of the clusters on bycatch, we suggest the use of transparent and objective tools. In particular, we discuss the use of Bayesian Belief Networks (BBNs) as a promising probabilistic modelling method for an evidence-based trade-off between removal efficiency and bycatch. We argue that BBN probabilistic models are a valuable tool to assist stakeholders, prior to the deployment of any clean-up technology, in selecting the best-suited mechanism to collect floating plastic debris while managing potential adverse effects on the ecosystem.


Defective mucosal movement at the gastroesophageal junction in patients with gastroesophageal reflux disease.
Little is known about the role of muscularis mucosa at the gastroesophageal junction (GEJ). To evaluate the movement of the mucosa/muscularis-mucosa/submucosa (MMS) at the GEJ in normal subjects and in patients with gastroesophageal reflux disease (GERD). Gastroesophageal junctions of 20 non-GERD subjects and 10 patients with GERD were evaluated during 5 mL swallows using two methods: in high-resolution endoluminal ultrasound and manometry, the change in the GEJ luminal pressures and cross-sectional area of esophageal wall layers were measured; in abdominal ultrasound, the MMS movement at the GEJ was analyzed. Endoluminal ultrasound: In the non-GERD subjects, the gastric MMS moved rostrally into the distal esophagus at 2.17 s after the bolus first reached the GEJ. In GERD patients, the gastric MMS did not move rostrally into the distal esophagus. The maximum change in cross-sectional area of gastroesophageal MMS in non-GERD subjects and in GERD patients was 289 % and 183%, respectively. Abdominal ultrasound: In non-GERD subjects, the gastric MMS starts to move rostrally significantly earlier and to a greater distance than muscularis propria (MP) after the initiation of the swallow (1.75 vs. 3.00 s) and (13.97 vs. 8.91 mm). In GERD patients, there is no significant difference in the movement of gastric MMS compared to MP (6.74 vs. 6.09 mm). The independent movement of the gastric MMS in GERD subjects was significantly less than in non-GERD subjects. In non-GERD subjects, the gastric MMS moves rostrally into the distal esophagus during deglutitive inhibition and forms a barrier. This movement of the MMS is defective in patients with GERD.


Cardiometabolic risk in polycystic ovary syndrome: a comparison of different approaches to defining the metabolic syndrome.
Polycystic ovary syndrome (PCOS) is associated with insulin resistance and features in common with the metabolic syndrome (MetS)--factors shown to predict cardiovascular risk and type 2 diabetes. We investigated the prevalence and characteristics of the MetS in PCOS by three definitions-World Health Organization (WHO), National Cholesterol Education Program Adult Treatment Panel III (NCEP-ATP-III) and International Diabetes Federation (IDF)--and compared that with the background population. Cross-sectional study of 168 women with PCOS and 883 age-matched controls from the Australian Diabetes, Obesity and Lifestyle (AusDiab) study. Prevalence of the MetS in PCOS subjects was 33% by WHO, 37% by NCEP-ATP-III and 40% by IDF criteria, compared with 10% by NCEP-ATP-III and 13% by IDF in controls (P < 0.001). MetS by WHO criteria was not calculated in the AusDiab population. Age was an independent predictor of MetS in PCOS and controls. The prevalence of MetS was significantly higher among those with PCOS (P = 0.027) in obese women (BMI > 30 kg/m(2)), and higher but not significantly so in overweight (BMI 25-30 kg/m(2)) women (P = 0.052). Dehydroepiandrosterone sulphate was associated with a lower risk of the MetS--Odds ratio 0.86 (95% confidence interval, 0.77-0.97, P = 0.011). An approximate 4-fold increase in the prevalence of the MetS in women with PCOS compared with the general population, consistent with the proposed major role of insulin and obesity in the syndrome, implies greater risk of cardiometabolic disease in women with PCOS. However, this estimate is likely to vary according to PCOS definition, ethnicity and different aetiological pathways to PCOS.


Heartland Virus Epidemiology, Vector Association, and Disease Potential.
First identified in two Missouri farmers exhibiting low white-blood-cell and platelet counts in 2009, Heartland virus (HRTV) is genetically closely related to severe fever with thrombocytopenia syndrome virus (SFTSV), a tick-borne phlebovirus producing similar symptoms in China, Korea, and Japan. Field isolations of HRTV from several life stages of unfed, host-seeking Amblyomma americanum, the lone star tick, implicated it as a putative vector capable of transstadial transmission. Laboratory vector competence assessments confirmed transstadial transmission of HRTV, demonstrated vertical infection, and showed co-feeding infection between A. americanum. A vertical infection rate of 33% from adult females to larvae in the laboratory was observed, while only one of 386 pools of molted nymphs (1930) reared from co-feeding larvae was positive for HRTV (maximum-likelihood estimate of infection rate = 0.52/1000). Over 35 human HRTV cases, all within the distribution range of A. americanum, have been documented. Serological testing of wildlife in areas near the index human cases, as well as in widely separated regions of the eastern United States where A. americanum occur, indicated many potential hosts such as raccoons and white-tailed deer. Attempts, however, to experimentally infect mice, rabbits, hamsters, chickens, raccoons, goats, and deer failed to produce detectable viremia. Immune-compromised mice and hamsters are the only susceptible models. Vertical infection augmented by co-feeding transmission could play a role in maintaining the virus in nature. A more complete assessment of the natural transmission cycle of HRTV coupled with serosurveys and enhanced HRTV disease surveillance are needed to better understand transmission dynamics and human health risks.


Microbial communities along biogeochemical gradients in a hydrocarbon-contaminated aquifer.
Micro-organisms are known to degrade a wide range of toxic substances. How the environment shapes microbial communities in polluted ecosystems and thus influences degradation capabilities is not yet fully understood. In this study, we investigated microbial communities in a highly complex environment: the capillary fringe and subjacent sediments in a hydrocarbon-contaminated aquifer. Sixty sediment sections were analysed using terminal restriction fragment length polymorphism (T-RFLP) fingerprinting, cloning and sequencing of bacterial and archaeal 16S rRNA genes, complemented by chemical analyses of petroleum hydrocarbons, methane, oxygen and alternative terminal electron acceptors. Multivariate statistics revealed concentrations of contaminants and the position of the water table as significant factors shaping the microbial community composition. Micro-organisms with highest T-RFLP abundances were related to sulphate reducers belonging to the genus Desulfosporosinus, fermenting bacteria of the genera Sedimentibacter and Smithella, and aerobic hydrocarbon degraders of the genus Acidovorax. Furthermore, the acetoclastic methanogens Methanosaeta, and hydrogenotrophic methanogens Methanocella and Methanoregula were detected. Whereas sulphate and sulphate reducers prevail at the contamination source, the detection of methane, fermenting bacteria and methanogenic archaea further downstream points towards syntrophic hydrocarbon degradation.


Development and validation of the supervisory relationship measure: a self-report questionnaire for use with supervisors.
To develop a valid and reliable measure of the supervisory relationship (SR) from the perspective of the supervisor. A cross-sectional self-report questionnaire design was used with a repeated component for a subsample. Qualitative data from a previous study of the SR (collected by interviews with supervisors) were used to construct an 89-item self-report questionnaire measure (Supervisory Relationship Measure [SRM]). This was administered to 267 clinical psychology supervisors involved in supervising UK trainee clinical psychologists. One hundred and thirty-four participants completed the measure a second time to establish its test-retest reliability. Principal components analysis was used. Five factors were identified and labelled 'safe base', 'supervisor commitment', 'trainee contribution', 'external influences', and 'supervisor investment'. Fifty-one items (comprising five subscales based on the factors identified) were selected for the final measure. Analyses revealed the SRM to have good internal consistency, good test-retest reliability, and good construct (convergent and divergent) validity. A preliminary test of predictive (statistical) validity (supervisor satisfaction with the relationship and trainee competence as perceived by the supervisor) yielded promising results. The SRM is a valid and reliable measure of the SR from the supervisor perspective. It predicts supervisor satisfaction and preliminary evidence suggests that it may also predict trainee competence (as perceived by the supervisor). The findings are consistent with current theory on the SR. The SRM is, therefore, a promising tool for use in research, clinical, and training settings. The supervisory relationship measure (SRM) is a new psychometrically sound measure of the supervisory relationship (SR) with clinical and research uses. Clinically, it is a useful and structured way for supervisors to provide constructive feedback to their trainees. It can be used in conjunction with the Supervisory Relationship Questionnaire (SRQ) (completed by the supervisee) to support a dyadic discussion about clinical supervision. Currently the measure is only validated on Clinical Psychology trainees and should be used with caution with qualified staff and other professional groups. Participants were predominantly white British supervisors who chose to take part in an internet-based study and thus the sample may not be fully representative of the population.


A conative educational model for an intervention program in obese youth.
Obesity in children has increased in recent years throughout the world and is associated with adverse health consequences. Early interventions, including appropriate pedagogy strategies, are important for a successful intervention program. The aim of this study was to assess changes in body mass index, the ability to perform sport activities, behavior in the classroom and academic performance following one year of a health-wellness intervention program in obese youth. The CEMHaVi program included 37 obese children (19 girls and 18 boys). Participants received an intervention program consisting of physical activity and health education. Assessment included body mass index, academic performance, classroom performance and ability to perform sport activities. Paired t tests were used to assess the effects of intervention, and chi square was used to assess inter-action between measures. Findings of the study suggest significant decrease in Z scores of Body Mass Index and an improvement of academic performance, classroom behavior and the ability to perform sport activities (p < 0.05). Chi square testing showed significant positive inter-actions between body mass index, classroom behavior and academic performance. Results following year one of CEMHaVi showed that a program of physical activity and health education had positive effects on obesity, behavior in the classroom and the ability to perform sport activities in obese adolescents. Significant inter-action in changes between variables was observed. Findings are important for designing intervention models to improve health in obese youth.


Higher versus Lower Continuous Renal Replacement Therapy Intensity in Critically ill Patients with Liver Dysfunction.
To study the association between higher versus lower continuous renal replacement therapy (CRRT) intensity and mortality in critically ill patients with combined acute kidney injury and liver dysfunction. Post-hoc analysis of patients with liver dysfunction (Sequential Organ Failure Assessment liver score ≥2 or diagnosis of liver failure/transplant) included in the Randomized Evaluation of Normal versus Augmented Level renal replacement therapy (RENAL) trial. Of 444 patients, 210 (47.3%) were randomized to higher intensity (effluent flow 40 mL/kg/h) and 234 (52.7%) to lower intensity (effluent flow 25 mL/kg/h) therapy. Overall, 79 and 86% of prescribed effluent flow was delivered in the higher-intensity and lower-intensity groups, respectively (p < 0.001). In total, 113 (54.1%) and 120 (51.3%) patients died in each group. On multivariable Cox regression analysis, we found no independent association between higher CRRT intensity and mortality (HR 0.93, 95% CI 0.70-1.24; p = 0.642). In RENAL patients with liver dysfunction, higher CRRT intensity was not associated with reduced mortality.


Bidi smoking and health.
Bidi, made of low-grade tobacco, is popular among rural folk and the urban poor, in the South Asian region. The objectives of this study were: (1) to assess the extent of bidi smoking in different countries; (2) to compare its smoke and health hazards with conventional cigarettes and; (3) to recommend preventive measures. English language articles were identified through a MEDLINE computerised search (1966-1998) using the term 'bidi', 'beedi', 'beedie', 'biri' and manually in order to collate all the available information and to present a descriptive epidemiological profile. The prevalence of bidi smoking was found to be 21-56% among men in South Asian countries with very little information available about women. Concentrations of nicotine, tar and other toxic agents in the smoke are higher for bidi than for other cigarettes. Bidi smoking is also considered to cause about 2-3 times greater nicotine and tar inhalation than do conventional cigarettes, due to the poor combustibility of the bidi wrapper and greater puff frequency needed to keep the bidi alight. It has also been shown to have greater odds ratios, although not statistically significant, for various cancerous and chronic conditions in comparison with other cigarettes. Since bidi is hand-rolled, workers employed in bidi factories are at risk of developing cancerous conditions due to exposure to tobacco dust and flakes. We suggest that anti-smoking campaigns should be directed to the bidi smokers of all South Asian countries. Most bidi smokers are illiterate and malnourished which makes them more vulnerable to smoking-related morbidity and mortality. At the same time, measures should be taken to make bidi less harmful and to produce bidi by machine to reduce exposure-related toxicity among bidi production workers.


Cannabinoids as Key Regulators of Inflammasome Signaling: A Current Perspective.
Inflammasomes are cytoplasmic inflammatory signaling protein complexes that detect microbial materials, sterile inflammatory insults, and certain host-derived elements. Inflammasomes, once activated, promote caspase-1-mediated maturation and secretion of pro-inflammatory cytokines, interleukin (IL)-1β and IL-18, leading to pyroptosis. Current advances in inflammasome research support their involvement in the development of chronic inflammatory disorders in contrast to their role in regulating innate immunity. Cannabis (marijuana) is a natural product obtained from the Cannabis sativa plant, and pharmacologically active ingredients of the plant are referred to as cannabinoids. Cannabinoids and cannabis extracts have recently emerged as promising novel drugs for chronic medical conditions. Growing evidence indicates the potent anti-inflammatory potential of cannabinoids, especially Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol (CBD), and synthetic cannabinoids; however, the mechanisms remain unclear. Several attempts have been made to decipher the role of cannabinoids in modulating inflammasome signaling in the etiology of chronic inflammatory diseases. In this review, we discuss recently published evidence on the effect of cannabinoids on inflammasome signaling. We also discuss the contribution of various cannabinoids in human diseases concerning inflammasome regulation. Lastly, in the milieu of coronavirus disease-2019 (COVID-19) pandemic, we confer available evidence linking inflammasome activation to the pathophysiology of COVID-19 suggesting overall, the importance of cannabinoids as possible drugs to target inflammasome activation in or to support the treatment of a variety of human disorders including COVID-19.


Myeloid Cell-associated Resistance to PD-1/PD-L1 Blockade in Urothelial Cancer Revealed Through Bulk and Single-cell RNA Sequencing.
To define dominant molecular and cellular features associated with PD-1/PD-L1 blockade resistance in metastatic urothelial cancer. We pursued an unbiased approach using bulk RNA sequencing data from two clinical trials to discover (IMvigor 210) and validate (CheckMate 275) pretreatment molecular features associated with resistance to PD-1/PD-L1 blockade in metastatic urothelial cancer. We then generated single-cell RNA sequencing (scRNA-seq) data from muscle-invasive bladder cancer specimens to dissect the cellular composition underlying the identified gene signatures. We identified an adaptive immune response gene signature associated with response and a protumorigenic inflammation gene signature associated with resistance to PD-1/PD-L1 blockade. The adaptive immune response:protumorigenic inflammation signature expression ratio, coined the 2IR score, best correlated with clinical outcomes, and was externally validated. Mapping these bulk gene signatures onto scRNA-seq data uncovered their underlying cellular diversity, with prominent expression of the protumorigenic inflammation signature by myeloid phagocytic cells. However, heterogeneity in expression of adaptive immune and protumorigenic inflammation genes was observed among single myeloid phagocytic cells, quantified as the myeloid single cell immune:protumorigenic inflammation ratio (Msc2IR) score. Single myeloid phagocytic cells with low Msc2IR scores demonstrated upregulation of proinflammatory cytokines/chemokines and downregulation of antigen presentation genes, were unrelated to M1 versus M2 polarization, and were enriched in pretreatment blood samples from patients with PD-L1 blockade-resistant metastatic urothelial cancer. The balance of adaptive immunity and protumorigenic inflammation in individual tumor microenvironments is associated with PD-1/PD-L1 resistance in urothelial cancer with the latter linked to a proinflammatory cellular state of myeloid phagocytic cells detectable in tumor and blood.See related commentary by Drake, p. 4139.


A Stair Ascent and Descent Controller for a Powered Ankle Prosthesis.
This paper presents a control system for a powered transtibial prosthesis that provides stair ascent and descent capability, as well as ability for user-controlled transitions between walking, standing, stair ascent, and stair descent. The control system was implemented on a powered prosthesis and evaluated on a single unilateral transtibial amputee subject. The ability of the prosthesis to provide appropriate functionality during stair ascent and descent was assessed by comparing gait kinematics and kinetics of the prosthesis to those of a passive dynamic elastic response prosthesis and those of a set of non-amputee subjects. Data from the assessment indicates that the powered prosthesis is able to provide some desirable stair ascent and stair descent characteristics, relative to the passive prosthesis.


Inflammation is associated with a decrease of lipogenic factors in omental fat in women.
Obesity is characterized by systemic low-grade inflammation in which adipose tissue, especially the omental depot, is thought to play a key role. We have previously shown that inflammation impairs 3T3-L1 preadipocyte cell line differentiation. To explore whether this interaction also takes place in vivo, the expression of several genes related to inflammation and adipocyte differentiation was assessed in human samples. Paired adipose tissue biopsies (from omental and subcutaneous depots) were obtained from 24 women: 6 lean normoglycemic and 18 obese volunteers with different glycemic states (normoglycemic, glucose-intolerant, or type 2 diabetic). The expression levels of CD14, IL-18, leptin, adiponectin, sterol regulatory element binding transcription factor 1 (SREBP1), peroxisome proliferator-activated receptor gamma (PPARgamma), pre-B-cell colony enhancing factor 1 (PBEF1) (or visfatin), glycerol-3-phosphate dehydrogenase 1 (soluble) (GPD1), lipoprotein lipase (LPL), fatty acid binding protein 4, adipocyte (FABP4), and hypoxia-inducible factor 1alpha were determined by quantitative real-time PCR. CD14 and IL-18 were overexpressed in omental adipose tissue compared with the subcutaneous depot, irrespective of the subject's obesity or diabetes status. A significant decrease of LPL, GPD1, and leptin expression was observed in omental tissue, and an inverse correlation between expression of CD14 and IL-18 and that of PPARgamma, LPL, and FABP4 was observed. The underexpression of omental lipogenic markers was more accentuated in the presence of glucose intolerance. Furthermore, adiponectin and SREBP1 expression was also significantly decreased in omental tissue of type 2 diabetic patients. PBEF1 and HIF1alpha expression remained comparable in all samples. Therefore, in humans, inflammation is increased in the omental depot, as evidenced by CD14 and IL-18 expression. In this localization, the inflammatory state is associated with a decreased expression of lipogenic markers, which is more pronounced in diabetic subjects.


Aortic microcalcification is associated with elastin fragmentation in Marfan syndrome.
Marfan syndrome (MFS) is a connective tissue disorder in which aortic rupture is the major cause of death. MFS patients with an aortic diameter below the advised limit for prophylactic surgery (<5 cm) may unexpectedly experience an aortic dissection or rupture, despite yearly monitoring. Hence, there is a clear need for improved prognostic markers to predict such aortic events. We hypothesize that elastin fragments play a causal role in aortic calcification in MFS, and that microcalcification serves as a marker for aortic disease severity. To address this hypothesis, we analysed MFS patient and mouse aortas. MFS patient aortic tissue showed enhanced microcalcification in areas with extensive elastic lamina fragmentation in the media. A causal relationship between medial injury and microcalcification was revealed by studies in vascular smooth muscle cells (SMCs); elastin peptides were shown to increase the activity of the calcification marker alkaline phosphatase (ALP) and reduce the expression of the calcification inhibitor matrix GLA protein in human SMCs. In murine Fbn1C1039G/+ MFS aortic SMCs, Alpl mRNA and activity were upregulated as compared with wild-type SMCs. The elastin peptide-induced ALP activity was prevented by incubation with lactose or a neuraminidase inhibitor, which inhibit the elastin receptor complex, and a mitogen-activated protein kinase kinase-1/2 inhibitor, indicating downstream involvement of extracellular signal-regulated kinase-1/2 (ERK1/2) phosphorylation. Histological analyses in MFS mice revealed macrocalcification in the aortic root, whereas the ascending aorta contained microcalcification, as identified with the near-infrared fluorescent bisphosphonate probe OsteoSense-800. Significantly, microcalcification correlated strongly with aortic diameter, distensibility, elastin breaks, and phosphorylated ERK1/2. In conclusion, microcalcification co-localizes with aortic elastin degradation in MFS aortas of humans and mice, where elastin-derived peptides induce a calcification process in SMCs via the elastin receptor complex and ERK1/2 activation. We propose microcalcification as a novel imaging marker to monitor local elastin degradation and thus predict aortic events in MFS patients. Copyright © 2017 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.


Functional genomic characterization of virulence factors from necrotizing fasciitis-causing strains of Aeromonas hydrophila.
The genomes of 10 Aeromonas isolates identified and designated Aeromonas hydrophila WI, Riv3, and NF1 to NF4; A. dhakensis SSU; A. jandaei Riv2; and A. caviae NM22 and NM33 were sequenced and annotated. Isolates NF1 to NF4 were from a patient with necrotizing fasciitis (NF). Two environmental isolates (Riv2 and -3) were from the river water from which the NF patient acquired the infection. While isolates NF2 to NF4 were clonal, NF1 was genetically distinct. Outside the conserved core genomes of these 10 isolates, several unique genomic features were identified. The most virulent strains possessed one of the following four virulence factors or a combination of them: cytotoxic enterotoxin, exotoxin A, and type 3 and 6 secretion system effectors AexU and Hcp. In a septicemic-mouse model, SSU, NF1, and Riv2 were the most virulent, while NF2 was moderately virulent. These data correlated with high motility and biofilm formation by the former three isolates. Conversely, in a mouse model of intramuscular infection, NF2 was much more virulent than NF1. Isolates NF2, SSU, and Riv2 disseminated in high numbers from the muscular tissue to the visceral organs of mice, while NF1 reached the liver and spleen in relatively lower numbers on the basis of colony counting and tracking of bioluminescent strains in real time by in vivo imaging. Histopathologically, degeneration of myofibers with significant infiltration of polymorphonuclear cells due to the highly virulent strains was noted. Functional genomic analysis provided data that allowed us to correlate the highly infectious nature of Aeromonas pathotypes belonging to several different species with virulence signatures and their potential ability to cause NF.


A partially folded intermediate species of the beta-sheet protein apo-pseudoazurin is trapped during proline-limited folding.
The folding of apo-pseudoazurin, a 123-residue, predominantly beta-sheet protein with a complex Greek key topology, has been investigated using several biophysical techniques. Kinetic analysis of refolding using far- and near-ultraviolet circular dichroism (UV CD) shows that the protein folds slowly to the native state with rate constants of 0.04 and 0.03 min(-1), respectively, at pH 7.0 and at 15 degrees C. This process has an activation enthalpy of approximately 90 kJ/mole and is catalyzed by cyclophilin A, indicating that folding is limited by trans-cis proline isomerization, presumably around the Xaa-Pro 20 bond that is in the cis isomer in the native state. Before proline isomerization, an intermediate accumulates during folding. This species has a substantial signal in the far-UV CD, a nonnative signal in the near-UV CD, exposed hydrophobic surfaces (judged by 1-anilino naphthalenesulphonate binding), a noncooperative denaturation transition, and a dynamic structure (revealed by line broadening on the nuclear magnetic resonance time scale). We compare the properties of this intermediate with partially folded states of other proteins and discuss its role in folding of this complex Greek key protein.


Novel metrics for relating personal heat exposure to social risk factors and outdoor ambient temperature.
A more precise understanding of individual-level heat exposure may be helpful to advance knowledge about heat-health impacts and effective intervention strategies, especially in light of projected increases in the severity and frequency of extreme heat events. We developed and interrogated different metrics for quantifying personal heat exposure and explored their association with social risk factors. To do so, we collected simultaneous personal heat exposure data from 64 residents of metropolitan Phoenix, Arizona. From these data, we derived five exposure metrics: Mean Individually Experienced Temperature (IET), Maximum IET, Longest Exposure Period (LEP), Percentage Minutes Above Threshold (PMAT), and Degree Minutes Above Threshold (DMAT), and calculated each for Day Hours, Night Hours, and All Hours of the study period. We then calculated effect sizes for the associations between those metrics and four social risk factors: neighborhood vulnerability, income, home cooling type, and time spent outside. We also investigated exposure misclassification by constructing linear regression models of observations from a regional weather station and hourly IET for each participant. Our analysis revealed that metric choice and timeframe added depth and nuance to our understanding of differences in exposure within and between populations. We found that time spent outside and income were the two risk factors most strongly associated with personal heat exposure. We also found evidence that Mean IET is a good, but perhaps not optimal, measure for assessing group differences in exposure. Most participants' IETs were poorly correlated with regional weather station observations and the slope and correlation coefficient for linear regression models between regional weather station data and IETs varied widely among participants. We recommend continued efforts to investigate personal heat exposure, especially in combination with physiological indicators, to improve our understanding of links between ambient temperatures, social risk factors, and health outcomes.


Airborne medicine: bacterial volatiles and their influence on plant health.
Like most other eukaryotes, plants do not live alone but in close association with a diverse microflora. These plant-associated microbes contribute to plant health in many different ways, ranging from modulation of hormonal pathways to direct antibiosis of plant pathogens. Over the last 15 yr, the importance of volatile organic compounds as mediators of mutualistic interactions between plant-associated bacteria and their hosts has become evident. This review summarizes current knowledge concerning bacterial volatile-mediated plant protection against abiotic and biotic stresses. It then discusses the translational potential of such metabolites or of their emitters for sustainable crop protection, the possible ways to harness this potential, and the major challenges still preventing us from doing so. Finally, the review concludes with highlighting the most pressing scientific gaps that need to be filled in order to enable a better understanding of: the molecular mechanisms underlying the biosynthesis of bacterial volatiles; the complex regulation of bacterial volatile emission in natural communities; the perception of bacterial volatiles by plants; and the modes of actions of bacterial volatiles on their host.


Linking outcomes from peabody picture vocabulary test forms using item response models.
The present work describes how vocabulary ability as assessed by 3 different forms of the Peabody Picture Vocabulary Test (PPVT; Dunn & Dunn, 1997) can be placed on a common latent metric through item response theory (IRT) modeling, by which valid comparisons of ability between samples or over time can then be made. Responses from 2,625 cases in a longitudinal study of 697 persons for 459 unique PPVT items (175 items from Peabody Picture Vocabulary Test--Revised [PPVT-R] Form M [Dunn & Dunn, 1981], 201 items from Peabody Picture Vocabulary Test--3 [PPVT-3] Form A [Dunn & Dunn, 1997], and 83 items from PPVT-3 Form B [Dunn & Dunn, 1997]) were analyzed using a 2-parameter logistic IRT model. The test forms each covered approximately ± 3 SDs of vocabulary ability with high reliability. Some differences between item sets in item difficulty and discrimination were found between the PPVT-3 Forms A and B. Comparable estimates of vocabulary ability obtained from different test forms can be created through IRT modeling. The authors have also written a freely available SAS program that uses the obtained item parameters to provide IRT ability estimates given item responses to any of the 3 forms. This scoring resource will allow others with existing PPVT data to benefit from this work as well.


Activation delay in healed myocardial infarction: a comparison between model and experiment.
Conduction delay in healed myocardial infarction, facilitating reentry, is frequently based on an increased path length the activation has to travel in a matrix of merging and diverging bundles that survive in the infarcted area. Additional delay occurs at sites where bundles bifurcate. The purpose of this study was to investigate conduction delay at sites where bundles bifurcate. A computer model was developed to simulate spread of activation in a two-dimensional sheet of excitable elements. A structure consisting of two isolated bundles merging into a single one was modeled. Extracellular electrograms calculated in the model were comparable to electrograms obtained in a superfused infarcted papillary muscle model. A zone of crowded isochrones or local conduction delay was found at the site where an isolated bundle bifurcated. The position of the isochrones in this area depended on the way activation times were determined. Lines of activation delay were mainly perpendicular to the fiber direction. In conclusion, the results have enabled us to better understand extracellular electrograms at pivoting points and show that activation sequences at a microscopic level can best be constructed on the basis of Laplacian signals.


Gold mining in the Peruvian Amazon: global prices, deforestation, and mercury imports.
Many factors such as poverty, ineffective institutions and environmental regulations may prevent developing countries from managing how natural resources are extracted to meet a strong market demand. Extraction for some resources has reached such proportions that evidence is measurable from space. We present recent evidence of the global demand for a single commodity and the ecosystem destruction resulting from commodity extraction, recorded by satellites for one of the most biodiverse areas of the world. We find that since 2003, recent mining deforestation in Madre de Dios, Peru is increasing nonlinearly alongside a constant annual rate of increase in international gold price (∼18%/yr). We detect that the new pattern of mining deforestation (1915 ha/year, 2006-2009) is outpacing that of nearby settlement deforestation. We show that gold price is linked with exponential increases in Peruvian national mercury imports over time (R(2) = 0.93, p = 0.04, 2003-2009). Given the past rates of increase we predict that mercury imports may more than double for 2011 (∼500 t/year). Virtually all of Peru's mercury imports are used in artisanal gold mining. Much of the mining increase is unregulated/artisanal in nature, lacking environmental impact analysis or miner education. As a result, large quantities of mercury are being released into the atmosphere, sediments and waterways. Other developing countries endowed with gold deposits are likely experiencing similar environmental destruction in response to recent record high gold prices. The increasing availability of satellite imagery ought to evoke further studies linking economic variables with land use and cover changes on the ground.


Q Fever.
Q fever is a zoonotic disease found throughout the world. It is caused by the intracellular gram-negative bacterium Coxiella burnetii. Infection by C. burnetii occurs primarily by inhalation of the aerosolized bacteria from birthing animals or contaminated dust. The bacterium is very resistant to drying and heat, and is considered highly endemic in the Middle East, where it is likely underdiagnosed. Special Operations Forces medical providers should be aware of this disease, which must be in the differential diagnosis of a patient who has a history of fever, elevated liver enzymes, pneumonia in its acute form, and endocarditis, especially in those with existing valvular heart disease in its chronic form.


Dissociation of acid-base effects on substrate accumulation and on delta pH in dog mitochondria.
The relationship between the pH gradient (delta pH) and substrate accumulation was examined in mitochondria from dog renal cortex. Mitochondria were incubated in media containing bicarbonate or nonbicarbonate buffers. Mitochondrial delta pH was at equilibrium after 2 min incubation but citrate accumulation in the matrix space was still increasing. With nonbicarbonate buffer in rotenone-inhibited mitochondria, citrate and alpha-ketoglutarate concentrations in the matrix did not vary between pH 7.5 and 7.1; delta pH decreased from 0.62 to 0.52 as medium pH fell. With decreasing bicarbonate concentration (from 40 to 10 mM) and constant CO2 tension, concentrations of citrate, alpha-ketoglutarate, malate, glutamate, glutamine, and formate increased; pyruvate accumulation was lower at 10 than at 25 mM bicarbonate; delta pH remained constant. When respiratory changes were produced, concentrations of citrate, alpha-ketoglutarate, malate, and glutamate increased as medium pH fell and CO2 tension increased; accumulation of pyruvate, glutamine, and formate was unaffected. delta pH fell from 0.48 to 0.39 as CO2 tension rose from 3 to 12%. In uninhibited mitochondria, 14CO2 formation from labeled citrate was greater with 10 than with 40 mM bicarbonate; this difference as well as the accumulation of citrate in the matrix was blocked by inhibition of the tricarboxylate carrier with 1,2,3-benzenetricarboxylate. These results dissociate effects of acid-base change on mitochondrial substrate accumulation from changes in delta pH. They suggest a direct, bicarbonate-dependent influence of pH on multiple mitochondrial substrate carriers. This phenomenon may play an important role in metabolic regulation in renal cortex.


Robust oil-core nanocapsules with hyaluronate-based shells as promising nanovehicles for lipophilic compounds.
The design of nanodelivery systems has been recently considered as a solution to the major challenge in pharmaceutical research - poor bioavailability of lipophilic drugs. Nanocapsules with liquid oil cores and shells based on amphiphilic polysaccharides were developed here as robust carriers of hydrophobic active compounds. A series of modified charged hyaluronates were synthesized and used as stabilizing shells ensuring also the biocompatibility of the nanocapsules that is crucial for applications related to the delivery of lipophilic drugs in vivo. Importantly, the oil nanodroplets were found to be stably suspended in water for at least 15 months without addition of low molar mass surfactants. Moreover, their size and stability may be tuned by varying the relative content of hydrophobic and hydrophilic groups in the hyaluronate derivatives as was confirmed by dynamic light scattering and nanoparticle tracking analysis as well as electron microscopy. In vivo studies demonstrated that hyaluronate-based nanocapsules accumulated preferentially in the liver as well as in the lungs. Moreover, their accumulation was dramatically potentiated in endotoxemic mice. In vitro studies showed that the nanocapsules were taken up by liver sinusoidal endothelial cells and by mouse lung vascular endothelial cells. Importantly, the capsules were found to be nontoxic in an acute oral toxicity experiment even at a dose of 2000 mg per kg b.w. Biocompatible hyaluronate-based nanocapsules with liquid cores described herein represent a promising and tunable nanodelivery system for lipophilic active compounds via both oral and intravenous administration.


Image matching technology in high power LED's eutectic welding.
As the integration packaging density of high-power LED (Light Emitting Diode) chip modules become higher and higher, the accuracy and speed of visual inspection require higher demands correspondingly. The accurate position matching of substrates and flip-chip LEDs is one of the key technologies in the automatic eutectic welding process. In this paper we propose a method based on image features to complete the matching of the substrates and the flip-chip LEDs. Firstly, the substrate images and the flip-chip images are pre-processed respectively to obtain binary images. Then we apply Hough transformation to detect straight lines on the binary images, and find out the main linear directions to trigger the mechanical arms to adjust the positions of the substrate and the chip initially. Thirdly, we use eight neighbors interconnected domain algorithm for the first time to locate notable features of the substrate, and pass the located information to the control system to trigger the mechanical arm to adjust the substrate for the second time. At the same time, projection algorithm is applied to locate the anode of the flip-chip to drive the mechanical arm to adjust the position of the flip-chip again. Finally, the position information is used to trigger the mechanical arm to accomplish the matching of the substrate and the flip-chip. The proposed method improves the speed of matching on the basis of the accuracy of matching, which achieves these requirements of real-time and high accuracy applications.


Evidence of Absorptive Function in vivo in a Neo-Formed Bio-Artificial Intestinal Segment Using a Rodent Model.
A promising therapeutic approach for intestinal failure consists in elongating the intestine with a bio-engineered segment of neo-formed autologous intestine. Using an acellular biologic scaffold (ABS), we, and others, have previously developed an autologous bio-artificial intestinal segment (BIS) that is morphologically similar to normal bowel in rodents. This neo-formed BIS is constructed with the intervention of naïve stem cells that repopulate the scaffold in vivo, and over a period of time, are transformed in different cell populations typical of normal intestinal mucosa. However, no studies are available to demonstrate that such BIS possesses functional absorptive characteristics necessary to render this strategy a possible therapeutic application. The aim of this study was to demonstrate that the BIS generated has functional absorptive capacity. Twenty male August × Copenhagen-Irish (ACI) rats were used for the study. Two-centimeter sections of ABS were transplanted in the anti-mesenteric border of the small bowel. Animals were studied at 4, 8, and 12 weeks post-engraftment. Segments of intestine with preserved vascular supply and containing the BIS were isolated and compared to intestinal segments of same length in sham control animals (n = 10). D-Xylose solution was introduced in the lumen of the intestinal segments and after 2 h, urine and blood were collected to evaluate D-Xylose levels. Quantitative analysis was performed using ELISA. Morphologic, ultrastructural, and indirect functional absorption analyses were also performed. We observed neo-formed intestinal tissue with near-normal mucosa post-implantation as expected from our previously developed model. Functional characteristics such as morphologically normal enterocytes (and other cell types) with presence of brush borders and preserved microvilli by electron microscopy, preserved water, and ion transporters/channels (by aquaporin and cystic fibrosis transmembrane conductance regulator (CFTR)) were also observed. The capacity of BIS containing neo-formed mucosa to increase absorption of d-Xylose in the blood compared to normal intestine was also confirmed. With this study, we demonstrated for the first time that BIS obtained from ABS has functional characteristics of absorption confirming its potential for therapeutic interventions.


Phosphoinositides: Regulators of Nervous System Function in Health and Disease.
Phosphoinositides, the seven phosphorylated derivatives of phosphatidylinositol have emerged as regulators of key sub-cellular processes such as membrane transport, cytoskeletal function and plasma membrane signaling in eukaryotic cells. All of these processes are also present in the cells that constitute the nervous system of animals and in this setting too, these are likely to tune key aspects of cell biology in relation to the unique structure and function of neurons. Phosphoinositides metabolism and function are mediated by enzymes and proteins that are conserved in evolution, and analysis of knockouts of these in animal models implicate this signaling system in neural function. Most recently, with the advent of human genome analysis, mutations in genes encoding components of the phosphoinositide signaling pathway have been implicated in human diseases although the cell biological basis of disease phenotypes in many cases remains unclear. In this review we evaluate existing evidence for the involvement of phosphoinositide signaling in human nervous system diseases and discuss ways of enhancing our understanding of the role of this pathway in the human nervous system's function in health and disease.


Targeting treatment-resistant breast cancer stem cells with FKBPL and its peptide derivative, AD-01, via the CD44 pathway.
FK506-binding protein like (FKBPL) and its peptide derivative, AD-01, have already shown tumor growth inhibition and CD44-dependent antiangiogenic activity. Here, we explore the ability of AD-01 to target CD44-positive breast cancer stem cells (BCSC). Mammosphere assays and flow cytometry were used to analyze the effect of FKBPL overexpression/knockdown and AD-01 treatment ± other anticancer agents on BCSCs using breast cancer cell lines (MCF-7/MDA-231/ZR-75), primary patient samples, and xenografts. Delays in tumor initiation were evaluated in vivo. The anti-stem cell mechanisms were determined using clonogenic assays, quantitative PCR (qPCR), and immunofluorescence. AD-01 treatment was highly effective at inhibiting the BCSC population by reducing mammosphere-forming efficiency and ESA(+)/CD44(+)/CD24(-) or aldehyde dehydrogenase (ALDH)(+) cell subpopulations in vitro and tumor initiation in vivo. The ability of AD-01 to inhibit the self-renewal capacity of BCSCs was confirmed; mammospheres were completely eradicated by the third generation. The mechanism seems to be due to AD-01-mediated BCSC differentiation shown by a significant decrease in the number of holoclones and an associated increase in meroclones/paraclones; the stem cell markers, Nanog, Oct4, and Sox2, were also significantly reduced. Furthermore, we showed additive inhibitory effects when AD-01 was combined with the Notch inhibitor, DAPT. AD-01 was also able to abrogate a chemo- and radiotherapy-induced enrichment in BCSCs. Finally, FKBPL knockdown led to an increase in Nanog/Oct4/Sox2 and an increase in BCSCs, highlighting a role for endogenous FKBPL in stem cell signaling. AD-01 has dual antiangiogenic and anti-BCSC activity, which will be advantageous as this agent enters clinical trial.


Betulinic acid shows anticancer activity against equine melanoma cells and permeates isolated equine skin in vitro.
Equine malignant melanoma (EMM) is a frequently occurring dermoepidermal tumor in grey horses. Currently available therapies are either challenging or inefficient. Betulinic acid (BA), a naturally occurring triterpenoid, is a promising compound for cancer treatment. To evaluate the potential of BA as a topical therapy for EMM, its anticancer effects on primary equine melanoma cells and dermal fibroblasts and its percutaneous permeation through isolated equine skin were assessed in vitro. BA showed antiproliferative and cytotoxic effects on both primary equine melanoma cells and fibroblasts in a time- and dose-dependent manner. The lowest half-maximal inhibitory concentrations were obtained 96 h after the beginning of drug exposure (12.7 μmol/L and 23.6 μmol/L for melanoma cells eRGO1 and MelDuWi, respectively, in cytotoxicity assay). High concentrations of the compound were reached in the required skin layers in vitro. BA is a promising substance for topical EMM treatment. Further clinical studies in horses are necessary to assess safety and antitumoral effects in vivo.


The genomic underpinnings of eukaryotic virus taxonomy: creating a sequence-based framework for family-level virus classification.
The International Committee on Taxonomy of Viruses (ICTV) classifies viruses into families, genera and species and provides a regulated system for their nomenclature that is universally used in virus descriptions. Virus taxonomic assignments have traditionally been based upon virus phenotypic properties such as host range, virion morphology and replication mechanisms, particularly at family level. However, gene sequence comparisons provide a clearer guide to their evolutionary relationships and provide the only information that may guide the incorporation of viruses detected in environmental (metagenomic) studies that lack any phenotypic data. The current study sought to determine whether the existing virus taxonomy could be reproduced by examination of genetic relationships through the extraction of protein-coding gene signatures and genome organisational features. We found large-scale consistency between genetic relationships and taxonomic assignments for viruses of all genome configurations and genome sizes. The analysis pipeline that we have called 'Genome Relationships Applied to Virus Taxonomy' (GRAViTy) was highly effective at reproducing the current assignments of viruses at family level as well as inter-family groupings into orders. Its ability to correctly differentiate assigned viruses from unassigned viruses, and classify them into the correct taxonomic group, was evaluated by threefold cross-validation technique. This predicted family membership of eukaryotic viruses with close to 100% accuracy and specificity potentially enabling the algorithm to predict assignments for the vast corpus of metagenomic sequences consistently with ICTV taxonomy rules. In an evaluation run of GRAViTy, over one half (460/921) of (near)-complete genome sequences from several large published metagenomic eukaryotic virus datasets were assigned to 127 novel family-level groupings. If corroborated by other analysis methods, these would potentially more than double the number of eukaryotic virus families in the ICTV taxonomy. A rapid and objective means to explore metagenomic viral diversity and make informed recommendations for their assignments at each taxonomic layer is essential. GRAViTy provides one means to make rule-based assignments at family and order levels in a manner that preserves the integrity and underlying organisational principles of the current ICTV taxonomy framework. Such methods are increasingly required as the vast virosphere is explored.


Knowledge, skills and attitudes of older people and staff about getting up from the floor following a fall: a qualitative investigation.
Falls are the most common reason for ambulance callouts resulting in non-conveyance. Even in the absence of injury, only half of those who fall can get themselves up off the floor, often remaining there over an hour, increasing risk of complications. There are recognized techniques for older people to learn how to get up after a fall, but these are rarely taught. The aim of this study was to develop an understanding of attitudes of older people, carers and health professionals on getting up following a fall. A qualitative focus group and semi-structured interviews were conducted with 28 participants, including community-dwelling older people with experience of a non-injurious fall, carers, physiotherapists, occupational therapists, paramedics and first responders. Data were transcribed and analysed systematically using the Framework approach. A stakeholder group of falls experts and service users advised during analysis. The data highlighted three areas contributing to an individual's capability to get-up following a fall: the environment (physical and social); physical ability; and degree of self-efficacy (attitude and beliefs about their own ability). These factors fell within the wider context of making a decision about needing help, which included what training and knowledge each person already had to manage their fall response. Staff described how they balance their responsibilities, prioritising the individual's immediate needs; this leaves limited time to address capability in the aforementioned three areas. Paramedics, routinely responding to falls, only receive training on getting-up techniques from within their peer-group. Therapists are aware of the skillset to breakdown the getting-up process, but, with limited time, select who to teach these techniques to. Neither therapists nor ambulance service staff routinely teach strategies on how to get up, meaning that healthcare professionals largely have a reactive role in managing falls. Interventions that address the environment, physical ability and self-efficacy could positively impact on peoples' capability to get up following a fall. Therefore, a more proactive approach would be to teach people techniques to manage these aspects of future falls and to provide them easily accessible information.


Iodane-Guided ortho C-H Allylation.
A metal-free C-H allylation strategy is described to access diverse functionalized ortho-allyl-iodoarenes. The method employs hypervalent (diacetoxy)iodoarenes and proceeds through the iodane-guided "iodonio-Claisen" allyl transfer. The use of allylsilanes bearing electron-withdrawing functional groups unlocks the functionalization of a broad range of substrates, including electron-neutral and electron-poor rings. The resulting ortho-allylated iodoarenes are versatile building blocks, with examples of downstream transformation including a concise synthesis of the experimental antimitotic core of Dosabulin. DFT calculations shed additional light on the reaction mechanism, with notable aspects including the aromatic character of the transition-state structure for the [3,3] sigmatropic rearrangement, as well as the highly stereoconvergent nature of the trans-product formation.


Antibodies against arylcyclohexylamines and their similarities in binding specificity with the phencyclidine receptor.
Rabbit antibodies were generated against five unique epitopes of phencyclidine (PCP)-like molecules to determine the molecular requirements for arylcyclohexylamine binding to the PCP receptor. Three of the haptens contained the three ring structures of PCP. A fourth hapten was synthesized from a derivative of the highly potent PCP analog, 1-[1-(2-thienyl)cyclohexyl]piperidine. The fifth hapten, 5-[N-(1'-phenylcyclohexyl)amino]pentanoic acid, was used as a haptenic model for N-ethyl-1-phenylcyclohexylamine, one of the most potent arylcyclohexylamines. These haptens were bound covalently to bovine serum albumin and were then used as antigens to immunize rabbits. The affinities and cross-reactivity patterns of the resulting five antibodies were studied in a [3H]PCP radioimmunoassay using standard curves of various arylcyclohexylamines. The dissociation constants ranged from 1.9 to 51.6 nM. From the average IC50 values of the radioimmunoassay dose-response curves, the relative potency of each ligand to PCP was determined. Least-squares linear regression was used to correlate these data with relative potency data from two [3H]PCP receptor binding assays and a PCP drug discrimination assay in the rat. Only relative potency data from the anti-5[N-(1'-phenylcyclohexyl)amino]pentanoic acid antibody showed a significant correlation with data from the three pharmacological studies (r2 = 0.80, 0.57 and 0.78, respectively; p less than .05 in all cases). These data indicated the 5-[N-(1'-phenylcyclohexyl)amino]pentanoic acid hapten contained the pharmacologically active features needed for arylcyclohexylamine binding to the PCP receptor.


Transcriptional profiling of ion channel genes in Brugada syndrome and other right ventricular arrhythmogenic diseases.
Brugada syndrome is an inherited sudden-death arrhythmia syndrome. Na(+)-current dysfunction is central, but mutations in the SCN5A gene (encoding the cardiac Na(+)-channel Nav1.5) are present in only 20% of probands. This study addressed the possibility that Brugada patients display specific expression patterns for ion-channels regulating cardiac conduction, excitability, and repolarization. Transcriptional profiling was performed on right-ventricular endomyocardial biopsies from 10 unrelated Brugada probands, 11 non-diseased organ-donors, seven heart-transplant recipients, 10 with arrhythmogenic right-ventricular cardiomyopathy, and nine with idiopathic right-ventricular outflow-tract tachycardia. Brugada patients showed distinct clustering differences vs. the two control and two other ventricular-tachyarrhythmia groups, including 14 of 77 genes encoding important ion-channel/ion-transporter subunits. Nav1.5 and K(+)-channels Kv4.3 and Kir3.4 were more weakly expressed, whereas the Na(+)-channel Nav2.1 and the K(+)-channel TWIK1 were more strongly expressed, in Brugada syndrome. Differences were also seen in Ca(2+)-homeostasis transcripts, including stronger expression of RYR2 and NCX1. The molecular profile of Brugada patients with SCN5A mutations did not differ from Brugada patients without SCN5A mutations. Brugada patients exhibit a common ion-channel molecular expression signature, irrespective of the culprit gene. This finding has potentially important implications for our understanding of the pathophysiology of Brugada syndrome, with possible therapeutic and diagnostic consequences.


Combinatorial optimization of a CD4-mimetic miniprotein and cocrystal structures with HIV-1 gp120 envelope glycoprotein.
Miniproteins provide a bridge between proteins and small molecules. Here we adapt methods from combinatorial chemistry to optimize CD4M33, a synthetic miniprotein into which we had previously transplanted the HIV-1 gp120 binding surface of the CD4 receptor. Iterative deconvolution of generated libraries produced CD4M47, a derivative of CD4M33 that had been optimized at four positions. Surface plasmon resonance demonstrated fourfold to sixfold improvement in CD4M47 affinity for gp120 to a level about threefold tighter than that of CD4 itself. Assessment of the neutralization properties of CD4M47 against a diverse range of isolates spanning from HIV-1 to SIVcpz showed that CD4M47 retained the extraordinary breadth of the parent CD4M33, but yielded only limited improvements in neutralization potencies. Crystal structures of CD4M47 and a phenylalanine variant ([Phe23]M47) were determined at resolutions of 2.4 and 2.6 A, in ternary complexes with HIV-1 gp120 and the 17b antibody. Analysis of these structures revealed a correlation between mimetic affinity for gp120 and overall mimetic-gp120 interactive surface. A correlation was also observed between CD4- and mimetic-induced gp120 structural similarity and CD4- and mimetic-induced gp120 affinity for the CCR5 coreceptor. Despite mimetic substitutions, including a glycine-to-(d)-proline change, the gp120 conformation induced by CD4M47 was as close or closer to the conformation induced by CD4 as the one induced by the parent CD4M33. Our results demonstrate the ability of combinatorial chemistry to optimize a disulfide-containing miniprotein, and of structural biology to decipher the resultant interplay between binding affinity, neutralization breadth, molecular mimicry, and induced affinity for CCR5.


Differential effects of high-frequency repetitive transcranial magnetic stimulation over ipsilesional primary motor cortex in cortical and subcortical middle cerebral artery stroke.
Facilitation of cortical excitability of the ipsilesional primary motor cortex (M1) may improve dexterity of the affected hand after stroke. The effects of 10 Hz repetitive transcranial magnetic stimulation (rTMS) over ipsilesional M1 on movement kinematics and neural activity were examined in patients with subcortical or cortical stroke. Twenty-nine patients with impaired dexterity after stroke (16 subcortical middle cerebral artery [MCA] strokes, 13 MCA strokes involving subcortical tissue and primary or secondary cortical sensorimotor areas) received 1 session of 10 Hz rTMS (5-second stimulation, 25-second break, 1,000 pulses, 80% of the resting motor threshold) applied over: 1) ipsilesional M1 and 2) vertex (control stimulation). For behavioral testing, 29 patients performed index finger and hand tapping movements with the affected and unaffected hand prior to and following each rTMS application. For functional magnetic resonance imaging, 18 patients performed index finger tapping movements with the affected and unaffected hand before and after each rTMS application. Ten-Hz rTMS over ipsilesional M1, but not over vertex, improved movement kinematics in 14 of 16 patients with subcortical stroke, but not in patients with additional cortical stroke. Ten-Hz rTMS slightly deteriorated dexterity of the affected hand in 7 of 13 cortical stroke patients. At a neural level, rTMS over ipsilesional M1 reduced neural activity of the contralesional M1 in 11 patients with subcortical stroke, but caused a widespread bilateral recruitment of primary and secondary motor areas in 7 patients with cortical stroke. Activity in ipsilesional M1 at baseline correlated with improvement of index finger tapping frequency induced by rTMS. The beneficial effects of 10 Hz rTMS over ipsilesional M1 on motor function of the affected hand depend on the extension of MCA stroke. Neural activity in ipsilesional M1 may serve as a surrogate marker for the effectiveness of facilitatory rTMS.


Ototoxicity of neomycin and polymyxin B following middle ear application in the chinchilla and baboon.
Previous experimental studies have demonstrated structural damage of the organ of Corti and stria vascularis following application of combination antibiotic otic drops to the middle ear. In this investigation the ototoxic effects of neomycin and polymyxin B (two antibiotics often used together in ototopical preparations) were separately evaluated after administration of each agent to the middle ear cavities of chinchillas and baboons. The antibiotics were administered in saline solution at the same concentrations used in Cortisporin Otic Suspension (3.5 mg/ml neomycin base, 10,000 units/ml polymyxin B). In both the rodent and primate, polymyxin B consistently produced greater cochlear damage than did neomycin. In fact, the extent of hair cell loss and strial injury produced by polymyxin B alone was, in many cases, comparable to that previously observed after application of Cortisporin Otic Suspension itself. Hair cell loss in the baboon was markedly less severe than in the chinchilla. It is believed that differences in position and structure of the round window membrane are important factors in the differing levels of ototoxicity observed in the rodent and primate.


The peroxisomal membrane protein Inp2p is the peroxisome-specific receptor for the myosin V motor Myo2p of Saccharomyces cerevisiae.
The faithful inheritance of organelles by daughter cells is essential to maintain the benefits afforded to eukaryotic cells by compartmentalization of biochemical functions. In Saccharomyces cerevisiae, the class V myosin, Myo2p, is involved in transporting different organelles, including the peroxisome, along actin cables to the bud. We identified Inp2p as the peroxisome-specific receptor for Myo2p. Cells lacking Inp2p fail to partition peroxisomes to the bud but are unaffected in the inheritance of other organelles. Inp2p is a peroxisomal membrane protein, preferentially enriched in peroxisomes delivered to the bud. Inp2p interacts directly with the globular tail of Myo2p. Cells overproducing Inp2p often transfer their entire populations of peroxisomes to buds. The levels of Inp2p oscillate with the cell cycle. Organelle-specific receptors like Inp2p explain how a single motor can move different organelles in distinct and specific patterns. To our knowledge, Inp2p is the first peroxisomal protein implicated in the vectorial movement of peroxisomes.


Hiding in plain sight; constrictive pericarditis.
We present a case of constrictive pericarditis that was especially difficult to diagnose. The patient presented with generalised oedema, dyspnoea and pleural effusions. History was significant for prior polysubstance abuse but was otherwise unremarkable. Physical examination revealed only jugular venous distention. CT demonstrated a normal pericardium with pleural effusions. Echocardiography showed mildly elevated right ventricular pressures with dyssynergic motion of the ventricular septum. No intervention was being carried out, but 1 month later further evaluation with a right and left heart catheterisation showed the classical square-root sign with equalisation of diastolic pressures in both ventricles as well as ventricular interdependence. Idiopathic constrictive pericarditis was thus diagnosed with a subsequent pericardial stripping which confirmed a thickened pericardium encasing the heart.


Effects of Facebook Comments on Attitude Toward Vaccines: The Roles of Perceived Distributions of Public Opinion and Perceived Vaccine Efficacy.
This study investigated if and how exposure to Facebook comments about vaccines influences one's attitude toward the vaccines. In this investigation, comments were examined in light of their effect on attitude toward vaccines through perceived distribution of public opinion on vaccines, and perceived vaccine efficacy was tested as a factor moderating relative effects of comments on perception of public opinion distributions. Results from an experimental study (N = 271) showed that exposure to a greater number of comments in a thread expressing (un)favorable opinions on the flu vaccine led to (un)favorable attitude toward the flu vaccine through a change in perceived distribution of public opinions on the vaccination. The indirect effect of comments on attitude toward the flu vaccine through perceived public opinion distributions was greater among participants with lower levels of perceived vaccine efficacy, while the direct effect of comments on attitude was not significant.


Predictors of poor sleep quality among Lebanese university students: association between evening typology, lifestyle behaviors, and sleep habits.
Adequate, good night sleep is fundamental to well-being and is known to be influenced by myriad biological and environmental factors. Given the unavailability of sleep data about Lebanon, the cultural shifts and socioeconomic pressures that have affected many aspects of society, particularly for students and working adults, as well as our understanding of sleep in university students in other countries, we conducted a national study to assess sleep quality and factors contributing to sleep and general health in a culture-specific context. A self-filled questionnaire, inquiring about sociodemographics, health-risk behaviors, personal health, and evaluating sleep quality and chronotype using standard scales was completed by 540 students at private and public universities in Lebanon. Overall, they reported sleeping 7.95±1.34 hours per night, although 12.3% reported sleeping <6.5 hours and more than half scored in the poor-sleeper category on the Pittsburgh Sleep Quality Index (PSQI). Sleep timing differed markedly between weekdays and weekends, with bedtimes and wake-up times delayed by 1.51 and 2.43 hours, respectively, on weekends. While most scored in the "neither type" category on the Morningness-Eveningness Questionnaire (MEQ), 24.5% were evening types and 7.3% were morning types. MEQ score was significantly correlated with smoking behavior and daily study onset, as well as with PSQI score, with eveningness associated with greater number of cigarettes, later study times, and poor sleep. We conclude that the prevalence of poor sleep quality among Lebanese university students is associated with reduced sleep duration and shifts in sleep timing between weekdays and weekends, especially among evening types. While chronotype and certain behavioral choices interact to affect sleep dimensions and quality, raising awareness about the importance of obtaining adequate nighttime sleep on daily performance and avoiding risky behaviors may help Lebanese students make better choices in school and work schedules.


Breast cancer and hormone replacement therapy: collaborative reanalysis of data from 51 epidemiological studies of 52,705 women with breast cancer and 108,411 women without breast cancer. Collaborative Group on Hormonal Factors in Breast Cancer.
The Collaborative Group on Hormonal Factors in Breast Cancer has brought together and reanalysed about 90% of the worldwide epidemiological evidence on the relation between risk of breast cancer and use of hormone replacement therapy (HRT). Individual data on 52,705 women with breast cancer and 108,411 women without breast cancer from 51 studies in 21 countries were collected, checked, and analysed centrally. The main analyses are based on 53,865 postmenopausal women with a known age at menopause, of whom 17,830 (33%) had used HRT at some time. The median age at first use was 48 years, and 34% of ever-users had used HRT for 5 years or longer. Estimates of the relative risk of breast cancer associated with the use of HRT were obtained after stratification of all analyses by study, age at diagnosis, time since menopause, body-mass index, parity, and the age a woman was when her first child was born. Among current users of HRT or those who ceased use 1-4 years previously, the relative risk of having breast cancer diagnosed increased by a factor of 1.023 (95% CI 1.011-1.036; 2p=0.0002) for each year of use; the relative risk was 1.35 (1.21-1.49; 2p=0.00001) for women who had used HRT for 5 years or longer (average duration of use in this group 11 years). This increase is comparable with the effect on breast cancer of delaying menopause, since among never-users of HRT the relative risk of breast cancer increases by a factor of 1.028 (95% CI 1.021-1.034) for each year older at menopause. 5 or more years after cessation of HRT use, there was no significant excess of breast cancer overall or in relation to duration of use. These main findings did not vary between individual studies. Of the many factors examined that might affect the relation between breast cancer risk and use of HRT, only a woman's weight and body-mass index had a material effect: the increase in the relative risk of breast cancer associated with long durations of use in current and recent users was greater for women of lower than of higher weight or body-mass index. There was no marked variation in the results according to hormonal type or dose but little information was available about long durations of use of any specific preparation. Cancers diagnosed in women who had ever used HRT tended to be less advanced clinically than those diagnosed in never-users. In North America and Europe the cumulative incidence of breast cancer between the ages of 50 and 70 in never-users of HRT is about 45 per 1000 women. The cumulative excess numbers of breast cancers diagnosed between these ages per 1000 women who began use of HRT at age 50 and used it for 5, 10, and 15 years, respectively, are estimated to be 2 (95% CI 1-3), 6 (3-9), and 12 (5-20). Whether HRT affects mortality from breast cancer is not known. The risk of having breast cancer diagnosed is increased in women using HRT and increases with increasing duration of use. This effect is reduced after cessation of use of HRT and has largely, if not wholly, disappeared after about 5 years. These findings should be considered in the context of the benefits and other risks associated with the use of HRT.


A genome-wide approach accounting for body mass index identifies genetic variants influencing fasting glycemic traits and insulin resistance.
Recent genome-wide association studies have described many loci implicated in type 2 diabetes (T2D) pathophysiology and β-cell dysfunction but have contributed little to the understanding of the genetic basis of insulin resistance. We hypothesized that genes implicated in insulin resistance pathways might be uncovered by accounting for differences in body mass index (BMI) and potential interactions between BMI and genetic variants. We applied a joint meta-analysis approach to test associations with fasting insulin and glucose on a genome-wide scale. We present six previously unknown loci associated with fasting insulin at P < 5 × 10(-8) in combined discovery and follow-up analyses of 52 studies comprising up to 96,496 non-diabetic individuals. Risk variants were associated with higher triglyceride and lower high-density lipoprotein (HDL) cholesterol levels, suggesting a role for these loci in insulin resistance pathways. The discovery of these loci will aid further characterization of the role of insulin resistance in T2D pathophysiology.


Stress and burnout among hemodialysis nurses: a single-center, prospective survey study.
Stress is a well known and identified problem within the nursing profession. Dialysis nurses are exposed to high level of stress. Increasing workload can aggravate stress and cause burnout and exhaustion. Stress and burnout are capable of having a detrimental impact on organizational productivity and pose serious health and safety hazards on the job. We aimed in this study to determine the type and level of stress and the amount of burnout among our dialysis nurses, and to evaluate the managing skills and the impact of stress on their work performance. There were 93 nurses (19 national and 74 expatriate nurses) who answered modified questionnaires to the aims of our prospective and descriptive correlational study. Our results show that most nurses involved in the study (national and expatriate) experienced a mild level of stress (79% and 68%, respectively) and moderate level of burnout (42% and 38%, respectively). The most common stressor among the national nurses was technical breakdowns of machines (15.9%) and that among expatriates was job insecurity (16.9%). The majority of the national nurses (21%) coped with this by increased sick leaves, whereas the majority (25%) of the expatriates responded by becoming easily frustrated. The most utilized coping skill among both groups was the relaxation methods (20.8% versus 24.9%) and the least utilized was denial (3.9% versus 0.5%). In conclusion, our results suggest the exposure of dialysis nurses to different types of stress and demonstrate the different experienced coping skills. These results may have implications for nursing management and hospital administration.


Interpretation of glucocorticoids in neonatal hair: a reflection of intrauterine glucocorticoid regulation?
Glucocorticoids (GCs) measured in neonatal hair might reflect intrauterine as well as postpartum GC regulation. We aimed to identify factors associated with neonatal hair GC levels in early life, and their correlation with maternal hair GCs. In a single-center observational study, mother-infant pairs (n = 107) admitted for >72 h at the maternity ward of a general hospital were included. At birth and an outpatient visit (OPV, n = 72, 44 ± 11 days postpartum), maternal and neonatal hair was analyzed for cortisol and cortisone levels by LC-MS/MS. Data were analyzed regarding: (1) neonatal GC levels postpartum and at the OPV, (2) associations of neonatal GC levels with maternal GC levels and (3) with other perinatal factors. (1) Neonatal GC levels were >5 times higher than maternal levels, with a decrease in ±50% between birth and the OPV for cortisol. (2) Maternal and neonatal cortisol, but not cortisone, levels were correlated both at postpartum and at the OPV. (3) Gestational age was associated with neonatal GC postpartum (log-transformed β (95% CI): cortisol 0.07 (0.04-0.10); cortisone 0.04 (0.01-0.06)) and at the OPV (cortisol 0.08 (0.04-0.12); cortisone 0.00 (-0.04 to 0.04)), while weaker associations were found between neonatal GCs and other perinatal and maternal factors. Neonatal hair GCs mainly reflect the third trimester increase in cortisol, which might be caused by the positive feedback loop, a placenta-driven phenomenon, represented by the positive association with GA. Between birth and 1.5 months postpartum, neonatal hair cortisol concentrations decrease sharply, but still appear to reflect both intra- and extrauterine periods.


Relation between complement activation and susceptibility to decompression sickness.
The consequences of complement activation and the symptoms of decompression sickness are similar. Consequently, the relation between the sensitivity of individuals to complement activation by air bubbles and their susceptibility to decompression sickness has been examined. Plasma samples from 34 individuals were incubated with air bubbles, and the concentration of the fluid phase metabolites of complement activation C3a, C4a, and C5a were measured with radioimmunoassays. It was found that both the anaphylatoxins C3a and C5a were produced by the presence of air bubbles but that the anaphylatoxin C4a was not. This finding indicates that air bubbles activate the complement system by the alternate pathway. One group of individuals was found to be particularly sensitive to complement activation by this pathway. They produced 3.3 times more C3a and 5.3 times more C5a in their plasma samples incubated with air bubbles as did the other group. Sixteen individuals were subjected to a series of pressure profiles that were severe enough to produce bubbles in their circulatory system that could be detected by Doppler ultrasonic monitoring. The group of individuals that had been identified as being more sensitive to complement activation by the alternate pathway was also found to be more susceptible to decompression sickness.


Plasma membrane calcium ATPases as novel candidates for therapeutic agent development.
Plasma membrane Ca2+ ATPases (PMCAs) are highly regulated transporters responsible for Ca2+ extrusion from all eukaryotic cells. Different PMCA isoforms are implicated in various tasks of Ca2+ regulation including bulk Ca2+ transport and localized Ca2+ signaling in specific membrane microdomains. Accumulating evidence shows that loss, mutation or inappropriate expression of different PMCAs is associated with pathologies ranging from hypertension, low bone density and male infertility to hearing loss and cerebellar ataxia. Compared to Ca2+ influx channels, PMCAs have lagged far behind as targets for drug development, mainly due to the lack of detailed understanding of their structure and specific function. This is rapidly changing thanks to integrated efforts combining biochemical, structural, cellular and physiological studies suggesting that selective modulation of PMCA isoforms may be of therapeutic value in the management of different and complex diseases. Both structurally informed rational design and high-throughput small molecule library screenings are promising strategies that are expected to lead to specific and isoform-selective modulators of PMCA function. This short review will provide an overview of the diverse roles played by PMCA isoforms in different cells and tissues and their emerging involvement in pathophysiological processes, summarize recent progress in obtaining structural information on the PMCAs, and discuss current and future strategies to develop specific PMCA inhibitors and activators for potential therapeutic applications.


Mechanochemical synthesis of three double perovskites: Cs2AgBiBr6, (CH3NH3)2TlBiBr6 and Cs2AgSbBr6.
A wide majority of the organic-inorganic hybrid perovskites employed in photovoltaics contain Pb, which is a negative issue due to its high toxicity and the low stability of the Pb-based three-dimensional (3D) perovskites. The double perovskites or "elpasolites" with the formula A2BB'X6 arise as an alternative to avoid the use of Pb, however, not many of the theoretically predicted structures have been synthesized so far due to several synthetic issues, such as, the formation of stable side products. Herein, we report the synthesis of three double perovskites, Cs2AgBiBr6, MA2TlBiBr6 and Cs2AgSbBr6, through a highly efficient and reproducible mechanochemical approach: the high energy ball milling. This synthetic approach does not require the use of organic solvents, so it is a greener method compared to those reported for other double perovskites. The Cs2AgBiBr6 and MA2TlBiBr6 double perovskites were synthesized with high purity as proved by X-ray diffraction (XRD) and X-ray fluorescence (XRF) measurements. However, the Cs2AgSbBr6 double perovskite was obtained in mixture with Cs3Sb2Br9, a side product of the reaction. Several attempts to prepare the Cs2AgSbBr6 double perovskite by using other synthetic methods have been unsuccessful due to the low formation enthalpy of the Cs3Sb2Br9 side product and only the hydrothermal method afforded Cs2AgSbBr6 in mixture with other compounds. We believe that the low temperature required in the ball milling synthesis is the key factor that allows the formation of the antimony double perovskite. Cs2AgSbBr6 is a brown powder with a bandgap energy of 1.93 eV as shown with diffuse reflectance measurements. The three powders exhibit a very high stability with no changes at all in the crystal structure after several months of storage at room temperature and ambient humidity.


The future is now: Age-progressed images motivate community college students to prepare for their financial futures.
Part of the challenge young people face when preparing for lifelong financial security is visualizing the far-off future. Age-progression technology has been shown to motivate young people to save for retirement. The current study applied age progression for motivating socioeconomically diverse community college students as part of a college planning course. We recruited 106 students enrolled in a mandatory "Transitioning to College" course and randomly assigned them to view age-progressed or same-aged digital avatars. Compared to controls, age-progressed participants gave more correct answers and exhibited higher confidence (i.e., fewer "don't know" responses) on a financial literacy test. Confidence mediated the effect of age progression on correct responses, but not the other way around, pointing to financial confidence as a precursor to effective financial education. Students also reported interest in attending more long-term financial planning workshops (e.g., investing and retirement) available through their college. No differences were observed in financial planning for the near term (e.g., student aid and credit cards). The current study demonstrates the viability of age progression as a practical, inexpensive, and scalable intervention. Findings also illustrate the significance of this intervention for reducing pervasive socioeconomic and age disparities in financial knowledge and enhancing long-term financial prospects across future generations. (PsycInfo Database Record (c) 2020 APA, all rights reserved).


High-resolution mapping of carbene-based protein footprints.
Carbene chemistry has been used recently in structural mass spectrometry as a labeling method for mapping protein surfaces. The current study presents a method for quantitating label distribution at the amino acid level and explores the nature and basis for an earlier observation of labeling bias. With the use of a method based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) applied to digests of holo-calmodulin, we developed a quantitation strategy to map site-specific incorporation of carbene, generated from photolysis of ionic label precursors 2-amino-4,4-azipentanoic acid and 4,4-azipentanoic acid. The approach provides reliable incorporation data for fragments generated by electron-transfer dissociation, whereas high-energy collisional dissociation leads to energy and sequence-dependent loss of the label as a neutral. However, both can produce data suitable for mapping residues in the interaction of holo-calmodulin with M13 peptide ligand. Site-specific labeling was monitored as a function of reagent, ionic strength, and temperature, demonstrating that electrostatic interactions at the protein surface can "steer" the distribution of label precursors to sites of surface charge and favor label insertion into residues in the vicinity of the surface charge. A further preference for insertion into carboxylates was observed, based on chemical reactivity. We suggest that decoupling surface partitioning from the chemistry of insertion offers a flexible, tunable labeling strategy for structural mass spectrometry that can be applied to a broad range of protein surface compositions and promotes the design of reagents to simplify the workflow.


Similarities and differences between aggressive and delinquent children and adolescents in a national sample.
To examine differences in the correlates, comorbidity and use of services between aggressive and delinquent children and adolescents. An Australian representative sample (n = 4083) of parents of children and adolescents were administered a psychiatric diagnostic interview, the Child Behaviour Checklist, and other instruments to measure service use. The characteristics of children with high scores (top 5%) in the aggressive and delinquent syndromes or both were then examined. The proportion of aggressive children decreased with increasing age while that of delinquents increased. The aggressive group was specifically associated with the impulsive-hyperactive subtype of attention deficit hyperactivity disorder (ADHD) (OR = 12.63; 95% CI = 5.97-26.74). Comorbidity between ADHD, aggression and delinquency was less frequent among adolescents than in children, with the exception of the inattentive subtype in which comorbidity was higher. Both aggressive and delinquent groups had a considerable overlap with conduct disorder. Aggressive and delinquent youths used services more often, but parents perceived aggressive children as more in need of help than delinquent ones. Living in a sole parent family was specifically associated with the delinquent group (OR = 3.34; 95% CI = 2.25-4.96). The results suggest that these empirically derived syndromes while sharing many features also differ in important ways, highlighting the need for further convergence between categorical and dimensional classifications. Their differential association with the subtypes of ADHD requires further examination and may help to understand the relationship between ADHD and conduct problems. The importance of aggressive behaviour in children should not be underestimated since it is associated with significant psychopathology, parental distress and use of services.


Familial adenomatous polyposis and the small bowel: a loco-regional review and current management strategies.
Small-bowel tumours are an important cause of morbidity and death in patients with familial adenomatous polyposis. Intensive endoscopic surveillance is now standard in the long-term management of this condition. Thus, lesions occurring throughout the small bowel are increasingly noted by oesophagogastroduodenoscopy and flexible pouchoscopy. Some occur commonly de novo (in stomach, duodenum and ampulla), while others may occur following surgery (polyps of the ileostomy, ileoanal pouch, or small bowel above an anastomosis). These differ widely in incidence, natural history and management. This review provides a regional overview of these lesions, in terms of current research findings and management protocols.


Bacterial load and cytokine profile in patients with cirrhosis following therapy with proton pump inhibitors: a prospective cohort study.
The aim of this study was to explore the presence of bacterial products and the cytokine profile in outpatients with cirrhosis before and after short-term (4-8 weeks) administration of proton pump inhibitors (PPIs). Seventeen patients with cirrhosis-male/female: 12/5; age: median 59.2 years (49-65); etiology: HBV±HDV 23.5%, HCV 17.7%, alcohol 41.2%, other 17.6%; Child-Pugh score: median 7.5 (5-12); Model for End-stage Liver Disease: 10.5 (7-21); ascites (%): 3 (17.7)-attending the outpatient clinics were included. None had hepatocellular carcinoma. Indications for PPIs were: esophagitis (n=6, 35.3%), peptic ulcer (n=10, 58.6%) and other (n=1, 5.9%). Bacterial DNA in serum and the levels of endotoxin, lipopolysaccharide binding protein, transforming growth factor-β, interleukin -1β, -6, -8, -12, -10, tumor necrosis factor-α and nitric oxide were assessed at baseline (time 1) and at the end of treatment (time 2). The Wilcoxon signed rank test was used to evaluate significant differences in the parameters assayed before and after PPI administration. No patients developed infection during the study period. Bacterial DNA was not detected before or after treatment. No significant differences were observed between the concentrations of any indices between times 1 and 2 (P>0.05). Subgroup analysis according to Child-Pugh stage yielded similar results. Short-term administration of PPIs had no effect on bacterial DNA, bacterial products or cytokine concentrations in patients with liver cirrhosis.


Down-regulation of habenular calcium-dependent secretion activator 2 induces despair-like behavior.
Calcium-dependent secretion activator 2 (CAPS2) regulates the trafficking and exocytosis of neuropeptide-containing dense-core vesicles (DCVs). CAPS2 is prominently expressed in the medial habenula (MHb), which is related to depressive behavior; however, how MHb neurons cause depressive symptoms and the role of CAPS2 remains unclear. We hypothesized that dysfunction of MHb CAPS neurons might cause defects in neuropeptide secretion and the activity of monoaminergic centers, resulting in depressive-like behaviors. In this study, we examined (1) CAPS2 expression in the habenula of depression animal models and major depressive disorder patients and (2) the effects of down-regulation of MHb CAPS2 on the animal behaviors, synaptic transmission in the interpeduncular nucleus (IPN), and neuronal activity of monoamine centers. Habenular CAPS2 expression was decreased in the rat chronic restraint stress model, mouse learned helplessness model, and showed tendency to decrease in depression patients who died by suicide. Knockdown of CAPS2 in the mouse habenula evoked despair-like behavior and a reduction of the release of DCVs in the IPN. Neuronal activity of IPN and monoaminergic centers was also reduced. These results implicate MHb CAPS2 as playing a pivotal role in depressive behavior through the regulation of neuropeptide secretion of the MHb-IPN pathway and the activity of monoaminergic centers.


A Wide-Field Optical Coherence Tomography Normative Database Considering the Fovea-Disc Relationship for Glaucoma Detection.
One purpose of this study was to collect wide-field swept-source optical coherence tomography (SS-OCT) data from healthy eyes and build a wide-filed normative database. Another purpose was to compare the glaucoma diagnostic ability of new parameters based on this normative database to the parameters that are currently in use, such as the peripapillary retinal nerve fiber layer (RNFL), macular ganglion cell-inner plexiform layer, and ganglion cell complex (GCC) thickness. This study had 220 healthy eyes and 292 eyes with early-stage glaucoma (EG) and moderate-stage glaucoma (MG) enrolled. Using the wide-field SS-OCT images (12 × 9 mm) of healthy eyes, a wide-field normative database was constructed by transforming and combining the individual images into a uniform template using the fovea and optic disc centers as fixed landmarks. Adjustment for the disc size was conducted. With this normative database, new parameters based on the ratio of the fovea-disc distance (FDD) consisting of the fovea-disc relationship were evaluated. The glaucoma diagnostic ability was assessed based on the area under the receiver operating characteristic curve (AUC). Among the new peripapillary parameters, the RNFL of the circumference of the circle with diameter 0.8 FDD showed the highest AUC value for EG and MG, but the value was not significantly superior to that of the initial RNFL (AUC = 0.940 vs. 0.937, P = 0.631). Among the macular parameters, the GCC of the area of the circle of 1.5 FDD showed the highest AUC value for EG and MG, and the value was significantly superior to that of initial GCC (AUC = 0.929 vs. 0.919, P = 0.033). However, there was no significant difference between the initial and adjusted GCC thickness in patients included in the EG or MG groups separately. A wide-field normative database was built to consider the relationship between the fovea and the optic disc. Considering this aspect, we found that the GCC analysis using a broader area presented a significantly greater glaucoma diagnostic performance for EG and MG in the macula than the initial parameter for the GCC. Based on this wide-field normative database, the clinical use of a wide-field deviation map may help diagnose the patients with EG and MG in the future.


Your Pillow May Not Guarantee a Good Night's Sleep or Symptom-Free Waking.
To describe the performance of the pillow that participants usually slept on with respect to retiring and waking cervico-thoracic symptoms, pillow comfort, and sleep quality. Participants (n=106) were systematically recruited for a field trial comparing their own pillow and five trial pillows. Participants provided daily retiring and waking symptom reports and sleep-quality and pillow-comfort ratings prospectively for 1 week on each pillow. Linear and logistic regression models were used to investigate the relationship between pillow use, age, gender, sleep quality, pillow comfort, and waking and temporal (overnight) symptom reports. No waking symptoms were reported by 42.5% of participants on their own pillow. Regular waking symptoms, failure to relieve retiring symptoms, uncomfortable pillows, and/or poor-quality sleep were reported by over 50% of participants. All participants who reported poor sleep quality also reported poor pillow comfort. Pillow-comfort reports were not related to any waking symptom report; however, reports of poor sleep quality were significantly related to waking cervical stiffness (adjusted odds ratio [AOR]=4.3 [Confidence Interval (CI): 1.3-15.6]) and scapula pain (AOR=6.1 [CI: 1.1-31.6]). Feather pillow users provided consistently low reports of pillow comfort and sleep quality. Many participants appear to have made poor pillow choices, as poor sleep quality, low pillow comfort, and waking symptoms were common. Further research is required to understand why people choose particular pillows to sleep on, as well as to identify the best fit between person and pillow to optimize sleep quality and reduce waking symptoms.


Confinement templates for hierarchical nanoparticle alignment prepared by azobenzene-based surface relief gratings.
Alignment of nanoparticles to hierarchical periodic structures is an emerging field in the development of patterned surfaces. Common alignment methods are based on templates that guide particle self-assembly. These can be formed using lithographic methods offering an almost free choice of the motif, while being expensive and time-consuming for large-scale production. Alternatively, template formation by controlled wrinkling offers a low-cost formation, but often suffers from the formation of defect structures like line-defects and cracks. Here, we show a preparation technique for nanoparticle alignment substrates that is based on the inscription of holographic surface relief gratings with a periodic sinusoidal wave pattern on the surface of azobenzene films. As interference patterns are employed for structure formation, very uniform and defect-free gratings with tunable grating height and grating period can be prepared. These substrates were successfully replicated to poly(dimethyl siloxane) and the replicas used for the alignment of polystyrene latex particles. Accordingly produced substrates exhibiting gratings with a variation in grating height allow for efficient screening of nanoparticle alignment in a geometrical confinement in one single experiment. We anticipate our studies as a promising tool for the development of sensors, tunable gratings and metamaterials.