Ethics in the minutiae: examining the role of the physical laboratory environment in ethical discourse.
Responsibility within life science research is a highly scrutinised field. Increasingly, scientists are presented with a range of duties and expectations regarding their conduct within the research setting. In many cases, these duties are presented deontologically, forgoing extensive discussion on how these are practically implemented into the minutiae of daily research practices. This de-contextualized duty has proven problematic when it comes to practical issues of compliance, however it is not often considered as a fundamental aspect of building ethics discourse. This paper examines this issue in detail, particularly focusing on how differences in the contrasts between the ideal and real physical research environments cause conceptual problems for scientists and retard ethical engagement. Such issues are particularly pertinent in low- and middle-income countries. This paper combines theoretical and empirical analyses using the concept of "dual-use" as a focalizing topic. The data show that the research environment acts as an intimate component in the interpretation and implementation of ethical actions.


The Impact of Starvation on the Microbiome and Gut-Brain Interaction in Anorexia Nervosa.
Interactions between the gut microbiome and the brain are of increasing interest to both researchers and clinicians. Evidence is mounting on the causal role of an altered gut microbiome in inflammatory diseases such as arthritis, inflammatory bowel disease, obesity and diabetes, and psychiatric diseases like anxiety and depression. Mechanisms include altered energy harvest from food, hormonal changes, increased gut permeability, inflammation, immune response, and a direct influence on the brain and behavior. Anorexia nervosa (AN) is the third most common disease in adolescence and exacts a high burden on patients and caregivers. It often becomes chronic and has the highest mortality of all psychiatric diseases. As AN is characterized by nutritional restrictions, weight loss, and severe behavioral symptoms including weight phobia, comorbid anxiety and depression, accompanied by endocrine alterations, increased inflammation, and immune response, exploring the role of the gut microbiome is crucial. Here, we present an overview of the potential mechanisms of interaction between the gut microbiome, the host and particularly the brain in AN and summarize the initial findings of microbiome research on AN. We conclude by identifying future research directions and potential therapeutic approaches, including nutritional interventions, probiotics, prebiotics and food supplements, that could become important additions to current AN therapy.


Resective epilepsy surgery for drug-resistant focal epilepsy: a review.
Epilepsy surgery is indicated for patients with focal seizures who do not respond to appropriate antiepileptic drug therapy consisting of 2 or more medications. To review resective surgery outcomes for focal epilepsy, to identify which patients benefit the most, and to discuss why epilepsy surgery may not be universally accepted. Medline and Cochrane databases were searched between January 1993 and June 2014 for randomized clinical trials, meta-analyses, systematic reviews, and large retrospective case series (>300 patients) using Medical Subject Headings and indexed text terms. Fifty-five articles were included. Subpopulations and prognostic factors were identified. Systematic reviews for cognitive, psychiatric, quality-of-life, and psychosocial outcomes were included. Two randomized clinical trials enrolling 118 patients with temporal lobe epilepsy found greater freedom from seizures with surgery when compared with continued medical treatment (58% vs 8% [n = 80] and 73% vs 0% [n = 38], P < .001). Nine systematic reviews and 2 large case series of medically refractory patients treated with surgery reported seizure-free outcomes in 34% to 74% of patients (median, 62.4%). The remainder of systematic reviews and meta-analyses examined subpopulations. Epilepsy surgery was less effective when there were extratemporal lesions, the epilepsy was not associated with a structural lesion, or both. Seizure-free outcomes were similar between children and adults. Hippocampal sclerosis and benign tumors were associated with better outcomes relative to other pathologies. Similar procedures such as selective amygdalohippocampectomy and temporal lobectomy for temporal lobe epilepsy were associated with subtle differences in seizure and neuropsychological outcome. There is low perioperative mortality (0.1%-0.5%) from epilepsy surgery. The most frequent neurologic complication is visual field defect occurring from temporal lobe resection. Quality of life improved after surgery but improved the most in patients who were seizure-free after surgery. Epilepsy surgery reduced seizure activity in randomized clinical trials when compared with continued medical therapy. Long-term cognitive, psychiatric, psychosocial, and quality-of-life outcomes were less well defined. Despite good outcomes from high-quality clinical trials, referrals of patients with seizures refractory to medical treatment remain infrequent.


Cognition-behaviour links in the persistence of panic.
The cognitive theory of panic disorder proposes that panic attacks occur as a result of an enduring tendency to misinterpret bodily sensations as a sign of imminent catastrophe such as a heart attack. The persistence of such catastrophic cognitions is in part due to the tendency of patients to avoid and/or escape situations where panic occurs. It is proposed that within-situation safety seeking behaviours also have the effect of maintaining catastrophic cognitions in the face of repeated panics during which the feared catastrophe does not occur. The association between catastrophic cognitions and within-situation safety behaviours matched the pattern predicted from the cognitive theory in 147 panic disorder patients. The implications of these findings for the way in which therapy is conducted are discussed.


A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast.
We have used a lipophilic styryl dye, N-(3-triethylammoniumpropyl)-4- (p-diethylaminophenyl-hexatrienyl) pyridinium dibromide (FM 4-64), as a vital stain to follow bulk membrane-internalization and transport to the vacuole in yeast. After treatment for 60 min at 30 degrees C, FM 4-64 stained the vacuole membrane (ring staining pattern). FM 4-64 did not appear to reach the vacuole by passive diffusion because at 0 degree C it exclusively stained the plasma membrane (PM). The PM staining decreased after warming cells to 25 degrees C and small punctate structures became apparent in the cytoplasm within 5-10 min. After an additional 20-40 min, the PM and cytoplasmic punctate staining disappeared concomitant with staining of the vacuolar membrane. Under steady state conditions, FM 4-64 staining was specific for vacuolar membranes; other membrane structures were not stained. The dye served as a sensitive reporter of vacuolar dynamics, detecting such events as segregation structure formation during mitosis, vacuole fission/fusion events, and vacuolar morphology in different classes of vacuolar protein sorting (vps) mutants. A particularly striking pattern was observed in class E mutants (e.g., vps27) where 500-700 nm organelles (presumptive prevacuolar compartments) were intensely stained with FM 4-64 while the vacuole membrane was weakly fluorescent. Internalization of FM 4-64 at 15 degrees C delayed vacuolar labeling and trapped FM 4-64 in cytoplasmic intermediates between the PM and the vacuole. The intermediate structures in the cytoplasm are likely to be endosomes as their staining was temperature, time, and energy dependent. Interestingly, unlike Lucifer yellow uptake, vacuolar labeling by FM 4-64 was not blocked in sec18, sec14, end3, and end4 mutants, but was blocked in sec1 mutant cells. Finally, using permeabilized yeast spheroplasts to reconstitute FM 4-64 transport, we found that delivery of FM 4-64 from the endosome-like intermediate compartment (labeled at 15 degrees C) to the vacuole was ATP and cytosol dependent. Thus, we show that FM 4-64 is a new vital stain for the vacuolar membrane, a marker for endocytic intermediates, and a fluor for detecting endosome to vacuole membrane transport in vitro.


Earliest amyloid and tau deposition modulate the influence of limbic networks during closed-loop hippocampal downregulation.
Research into hippocampal self-regulation abilities may help determine the clinical significance of hippocampal hyperactivity throughout the pathophysiological continuum of Alzheimer's disease. In this study, we aimed to identify the effects of amyloid-β peptide 42 (amyloid-β42) and phosphorylated tau on the patterns of functional connectomics involved in hippocampal downregulation. We identified 48 cognitively unimpaired participants (22 with elevated CSF amyloid-β peptide 42 levels, 15 with elevated CSF phosphorylated tau levels, mean age of 62.705 ± 4.628 years), from the population-based 'Alzheimer's and Families' study, with baseline MRI, CSF biomarkers, APOE genotyping and neuropsychological evaluation. We developed a closed-loop, real-time functional MRI neurofeedback task with virtual reality and tailored it for training downregulation of hippocampal subfield cornu ammonis 1 (CA1). Neurofeedback performance score, cognitive reserve score, hippocampal volume, number of apolipoprotein ε4 alleles and sex were controlled for as confounds in all cross-sectional analyses. First, using voxel-wise multiple regression analysis and controlling for CSF biomarkers, we identified the effect of healthy ageing on eigenvector centrality, a measure of each voxel's overall influence based on iterative whole-brain connectomics, during hippocampal CA1 downregulation. Then, controlling for age, we identified the effects of abnormal CSF amyloid-β42 and phosphorylated tau levels on eigenvector centrality during hippocampal CA1 downregulation. Across subjects, our main findings during hippocampal downregulation were: (i) in the absence of abnormal biomarkers, age correlated with eigenvector centrality negatively in the insula and midcingulate cortex, and positively in the inferior temporal gyrus; (ii) abnormal CSF amyloid-β42 (<1098) correlated negatively with eigenvector centrality in the anterior cingulate cortex and primary motor cortex; and (iii) abnormal CSF phosphorylated tau levels (>19.2) correlated with eigenvector centrality positively in the ventral striatum, anterior cingulate and somatosensory cortex, and negatively in the precuneus and orbitofrontal cortex. During resting state functional MRI, similar eigenvector centrality patterns in the cingulate had previously been associated to CSF biomarkers in mild cognitive impairment and dementia patients. Using the developed closed-loop paradigm, we observed such patterns, which are characteristic of advanced disease stages, during a much earlier presymptomatic phase. In the absence of CSF biomarkers, our non-invasive, interactive, adaptive and gamified neuroimaging procedure may provide important information for clinical prognosis and monitoring of therapeutic efficacy. We have released the developed paradigm and analysis pipeline as open-source software to facilitate replication studies.


Feedback control of the rate of peat formation.
The role of peatlands in the global carbon cycle is confounded by two inconsistencies. First, peatlands have been a large reservoir for carbon sequestered in the past, but may be either net sources or net sinks at present. Second, long-term rates of peat accumulation (and hence carbon sequestration) are surprisingly steady, despite great variability in the short-term rates of peat formation. Here, we present a feedback mechanism that can explain how fine-scale and short-term variability in peat-forming processes is constrained to give steady rates of peat accumulation over longer time-scales. The feedback mechanism depends on a humpbacked relationship between the rate of peat formation and the thickness of the aerobic surface layer (the acrotelm), such that individual microforms (hummocks, lawns, hollows and pools) expand or contract vertically in response to fluctuations in the position of the water table. Hummocks (but not hollows) 'evolve' to a steady state where changes in acrotelm thickness compensate for climate-mediated variations in surface wetness. With long-term growth of a topographically confined peat deposit, the steady state gradually shifts to a thicker acrotelm (i.e. taller hummocks) and lower rates of peat formation and carbon sequestration.


Structural Stability, Vibrational Properties, and Photoluminescence in CsSnI3 Perovskite upon the Addition of SnF2.
The CsSnI3 perovskite and the corresponding SnF2-containing material with nominal composition CsSnI2.95F0.05 were synthesized by solid-state reactions and structurally characterized by powder X-ray diffraction. Both materials undergo rapid phase transformation upon exposure to air from the black orthorhombic phase (B-γ-CsSnI3) to the yellow orthorhombic phase (Y-CsSnI3), followed by irreversible oxidation into Cs2SnI6 within several hours. The phase transition occurs at a significantly lower rate in the SnF2-containing material rather than in the pure perovskite. The high hole-carrier concentration of the materials prohibits the detection of Raman signals for B-γ-CsSnI3 and induces a very strong plasmonic reflectance in the far-IR. In contrast, far-IR phonon bands and a rich Raman spectrum are observed for the Y-CsSnI3 modification below 140 cm-1 with weak frequency shift gradients versus temperatures between -95 and +170 °C. Above 170 °C, the signal is lost due to B-α-CsSnI3 re-formation. The photoluminescence spectra exhibit residual blue shifts and broadening as a sign of structural transformation initiation.


Immunogenicity of tumor peptides: importance of peptide length and stability of peptide/MHC class II complex.
Nonameric P815AB, a cytotoxic-T-lymphocyte-defined minimal core peptide encoded by the murine mastocytoma gene P1A, fails to initiate CD4+ cell-dependent reactivity in vivo to class-I-restricted epitopes when mice are administered peptide-pulsed dendritic cells. Effective immunization requires T helper effects, such as those mediated by coimmunization with class-II-restricted (helper) peptides or by the use of recombinant interleukin-12 (rIL-12). Although P815AB does possess class-II-restricted epitopes, they are likely suboptimal, resulting in poor affinity and/or stability of MHC/P815AB complexes and inadequate activation of the antigen-presenting cell function of dendritic cells. The present study has examined a series of longer, P815AB-centered peptides (11-14 amino acids in length, all P1A-encoded) for their ability to initiate CD4+ and CD8+ cell-mediated responses to the nonamer in vivo, their ability to bind class II MHC in vitro, and their ability to assemble class II molecules stably. By means of a class-I-restricted skin test assay in mice receiving peptide-pulsed dendritic cells, we found that a 12-mer and a 13-mer effectively immunized against the core P815AB peptide, and that this correlated with IL-2 production in vitro by CD4+ cells in response to the nonamer. In vitro studies, involving affinity-purified class II molecules, showed that the capacity to assemble class II molecules stably, more than the affinity for class II MHC, correlated with the ability of the different P815AB peptides to prime the host to the core peptide seen by the T cells.


The health benefits of yoga and exercise: a review of comparison studies.
Exercise is considered an acceptable method for improving and maintaining physical and emotional health. A growing body of evidence supports the belief that yoga benefits physical and mental health via down-regulation of the hypothalamic-pituitary-adrenal (HPA) axis and the sympathetic nervous system (SNS). The purpose of this article is to provide a scholarly review of the literature regarding research studies comparing the effects of yoga and exercise on a variety of health outcomes and health conditions. Using PubMed((R)) and the key word "yoga," a comprehensive search of the research literature from core scientific and nursing journals yielded 81 studies that met inclusion criteria. These studies subsequently were classified as uncontrolled (n = 30), wait list controlled (n = 16), or comparison (n = 35). The most common comparison intervention (n = 10) involved exercise. These studies were included in this review. In the studies reviewed, yoga interventions appeared to be equal or superior to exercise in nearly every outcome measured except those involving physical fitness. The studies comparing the effects of yoga and exercise seem to indicate that, in both healthy and diseased populations, yoga may be as effective as or better than exercise at improving a variety of health-related outcome measures. Future clinical trials are needed to examine the distinctions between exercise and yoga, particularly how the two modalities may differ in their effects on the SNS/HPA axis. Additional studies using rigorous methodologies are needed to examine the health benefits of the various types of yoga.


The Schwartz Centre Rounds: Supporting mental health workers with the emotional impact of their work.
In healthcare settings, there is an emotional cost to caring which can result in compassion fatigue, burnout, secondary trauma, and compromised patient care. Innovative workplace interventions such as the Schwartz Rounds offer a group reflective practice forum for clinical and non-clinical professionals to reflect on the emotional aspects of working in health care. Whilst the Rounds are established in medical health practice, this study presents an evaluation of the Rounds offered to mental health services. The Rounds were piloted amongst 150 mental health professionals for 6 months and evaluated using a mixed-methods approach with standardized evaluation forms completed after each Round and a focus group (n = 9) at one-month follow-up. This paper also offers a unique six-year follow-up of the evaluation of the Rounds. Rounds were rated as helpful, insightful, and relevant, and at six years follow-up, Rounds were still rated as valuable and viewed as embedded. Focus groups indicated that Rounds were valued because of the opportunity to express emotions (in particular negative emotions towards patients that conflict with the professional care-role), share experiences, and feel validated and supported by colleagues. The findings indicate that Schwartz Rounds offer a positive application in mental healthcare settings. The study supports the use of interventions which provide an ongoing forum in which to discuss emotions, develop emotional literacy, provide peer support and set an intention for becoming a more compassionate organization in which to work.


Massive photothermal trapping and migration of particles by a tapered optical fiber.
A simple but highly efficient method for particles or bacteria trapping and removal from water is of great importance for local water purification, particularly, for sanitation. Here, we report a massive photothermal trapping and migration of dielectric particles (SiO2, 2.08-µm diameter) in water by using a tapered optical fiber (3.1-µm diameter for taper). With a laser beam of 1.55 µm (170 mW) injected into the fiber, particles moved towards the position, which is about 380 µm away from the tip of the fiber, and assembled at a 290 µm × 100 µm spindle-shaped region. The highest assembly speed of particles is 22.1 ind./s and the highest moving velocity is 20.5 µm/s, which were induced by both negative photophoresis and temperature gradient. The number of assembled particles can reach 10,150 in 15 minutes. With a move of the fiber, the assembled particles will also migrate. We found that, when the fiber was moved 172 µm away from its original location, almost all of the assembled 10,150 particles were migrated to a new location in 140 s with a distance of 172 µm from their original location.


2011 clinical practice guidelines for type 2 diabetes in Korea.
As in other countries, type 2 diabetes is major health concern in Korea. A dramatic increase in the prevalence of type 2 diabetes and its chronic complications has led to an increase in health costs and economic burdens. Early detection of high risk individuals, hidden diabetic patients, and improvement in the quality of care for the disease are the first steps to mitigate the increase in prevalence. The Committee of Clinical Practice Guidelines of the Korean Diabetes Association revised and updated the '3rd Clinical Practice Guidelines' at the end of 2010. In the guidelines, the committee recommended active screening of high risk individuals for early detection and added the hemoglobin A1c level to the diagnostic criteria for type 2 diabetes based on clinical studies performed in Korea. Furthermore, the committee members emphasized that integrating patient education and self-management is an essential part of care. The drug treatment algorithm based on the degree of hyperglycemia and patient characteristics were also updated.


Seasonality and synchrony of reproduction in three species of nectarivorous Philippines bats.
Differences among species and among years in reproductive seasonality (the tendency for clusters of events to fall at approximately the same point in each year) and synchrony (amount of clustering of events within a year) have been intensively studied in bats, but are difficult to assess. Here, we use randomization methods with circular statistics to test for synchrony and seasonality of reproduction in three species of nectarivorous megachiropteran bats on Negros Island in the central Philippines. In Rousettus amplexicaudatus, estimated dates of birth were both highly synchronous and highly seasonal. In Macroglossus minimus, estimated births were seasonal and significantly clustered within years, but within each year births occurred over a broad period, indicating a low level of synchrony. In Eonycteris spelaea, estimated births were also seasonal and had statistically significant synchrony, with birth periods within years intermediate in synchrony between R. amplexicaudatus and M. minimus. All three species had a similar seasonal pattern, with two birth periods in each year, centered on March or April and August or September. In one species, R. amplexicaudatus, primigravid females (in their first pregnancy) produced their young in June and July, a birth period significantly different in timing from the two birth periods of older adult females. This more conservative pattern of young females may allow higher survival of parents and offspring at cost of a lost reproductive opportunity. There was weak evidence that in some years primigravid females of M. minimus might differ in timing from older adults. There were few significant differences in reproductive timing among different years, and those differences were generally less than two weeks, even during a severe drought in the severe el Niño of 1983. The results suggest that these species follow an obligately seasonal pattern of reproductive timing with very little phenotypic plasticity. The resampling methods were sensitive to differences in timing of under two weeks, in some cases, suggesting that these are useful methods for analyses of seasonality in wild populations of bats.


Experience With the Pipeline Embolization Device for Posterior Circulations Aneurysms: A Multicenter Cohort Study.
The Pipeline Embolization Device (PED; Medtronic) has been used off-label for the treatment of challenging posterior circulation aneurysms. Data on this modality are primarily limited to small retrospective single-center series. To assess safety and efficacy of this treatment by establishing an international, multicenter collaboration. Consecutive posterior circulation aneurysms treated with the PED from 2012 to 2019 across 11 neurovascular centers were retrospectively reviewed. Baseline demographics, aneurysm and treatment characteristics, complications, occlusion status, and functional outcome were assessed. There were 149 posterior circulation aneurysms treated with PED in 146 patients. A total of 24 (16.4%) patients presented with subarachnoid hemorrhage. Most aneurysms were dissecting/blister (36.2%) in morphology, followed by saccular (35.6%) and fusiform (28.2%). The most common locations were the vertebral (51.7%) and basilar arteries (22.8%). Complete or near-complete occlusion (>90%) was achieved in 90.9% of aneurysms at a median follow-up of 12 mo. Dissecting/blister aneurysms were most likely to occlude (P = .06). Symptomatic neurologic complications occurred in 9.4% of aneurysms, associated with larger size, ruptured presentation, presentations with brain stem compression, cranial nerve palsy, or stroke. Favorable functional outcome (modified Rankin Score 0-2) was achieved in 86.2% of patients. There were 6 fatalities of which 4 occurred in aneurysmal subarachnoid hemorrhage patients. This multicenter study shows that PED for the treatment of posterior circulation is preferentially used for the treatment of fusiform and dissecting/blister aneurysm morphologies. Despite the challenges presented by these less-common morphologies, flow diversion may be performed with a neurologic complication rate of about 10% and favorable long-term aneurysm occlusion rates.


Early implementation of continuous renal replacement therapy optimizes casualty evacuation for combat-related acute kidney injury.
The purpose of this report was to review the initial use and feasibility of continuous renal replacement therapy (CRRT) among combat casualties in a war zone. Although rapid evacuation to more advanced levels of care has emerged as the standard approach, life-threatening sequelae of acute kidney injury (AKI) can preclude safe patient evacuation. For the first time in US combat casualty care, a sustained, intensivist-led CRRT program was initiated during 2010 at an Air Force theater hospital. A prospective study of consecutive US service members (USSMs) who developed combat-related renal failure and underwent CRRT at the Craig Joint Theater Hospital was undertaken. Baseline patient characteristics, indications for CRRT, laboratory values, and outcomes were evaluated. Nine USSMs were treated during 14-months. All were male, with a mean (SD) age of 28 (7) years and mean (SD) Injury Severity Score (ISS) of 34 (12). The dominant mechanism was blast injury (8 of 9), followed by gunshot wound (1 of 9). Most patients were Acute Kidney Injury Network (AKIN) 3 and all developed critical hyperkalemia (mean [SD], peak K⁺ 6.4 [0.4]). The peak plasma creatinine ranged from 1.4 mg/dL to 4.2 mg/dL (mean [SD], 3.3 [0.9] mg/dL). Patients had a mean (SD) of 17.6 [8.1] hours of CRRT before evacuation to higher echelons of care. All USSMs survived to achieve safe evacuation from the combat zone to the regional trauma center in Landstuhl, Germany (Landstuhl Regional Medical Center). Three patients died of multiorgan failure at Landstuhl Regional Medical Center. Six patients survived to undergo additional treatment in the United States. Intensivist-led CRRT is an effective therapeutic adjunct in the treatment of combat-related AKI. Provision of this extracorporeal therapy provides physiologic stabilization of casualties who might otherwise succumb to the sequelae of combat-related renal failure. These findings suggest that a self-sustaining CRRT program can be successfully implemented in combat support hospitals.


Conditional telomerase induction causes proliferation of hair follicle stem cells.
TERT, the protein component of telomerase, serves to maintain telomere function through the de novo addition of telomere repeats to chromosome ends, and is reactivated in 90% of human cancers. In normal tissues, TERT is expressed in stem cells and in progenitor cells, but its role in these compartments is not fully understood. Here we show that conditional transgenic induction of TERT in mouse skin epithelium causes a rapid transition from telogen (the resting phase of the hair follicle cycle) to anagen (the active phase), thereby facilitating robust hair growth. TERT overexpression promotes this developmental transition by causing proliferation of quiescent, multipotent stem cells in the hair follicle bulge region. This new function for TERT does not require the telomerase RNA component, which encodes the template for telomere addition, and therefore operates through a mechanism independent of its activity in synthesizing telomere repeats. These data indicate that, in addition to its established role in extending telomeres, TERT can promote proliferation of resting stem cells through a non-canonical pathway.


Experimental study on foam coverage on simulated longwall roof.
Testing was conducted to determine the ability of foam to maintain roof coverage in a simulated longwall mining environment. Approximately 27 percent of respirable coal mine dust can be attributed to longwall shield movement, and developing controls for this dust source has been difficult. The application of foam is a possible dust control method for this source. Laboratory testing of two foam agents was conducted to determine the ability of the foam to adhere to a simulated longwall face roof surface. Two different foam generation methods were used: compressed air and blower air. Using a new imaging technology, image processing and analysis utilizing ImageJ software produced quantifiable results of foam roof coverage. For compressed air foam in 3.3 m/s (650 fpm) ventilation, 98 percent of agent A was intact while 95 percent of agent B was intact on the roof at three minutes after application. At 30 minutes after application, 94 percent of agent A was intact while only 20 percent of agent B remained. For blower air in 3.3 m/s (650 fpm) ventilation, the results were dependent upon nozzle type. Three different nozzles were tested. At 30 min after application, 74 to 92 percent of foam agent A remained, while 3 to 50 percent of foam agent B remained. Compressed air foam seems to remain intact for longer durations and is easier to apply than blower air foam. However, more water drained from the foam when using compressed air foam, which demonstrates that blower air foam retains more water at the roof surface. Agent A seemed to be the better performer as far as roof application is concerned. This testing demonstrates that roof application of foam is feasible and is able to withstand a typical face ventilation velocity, establishing this technique's potential for longwall shield dust control.


Quantifying the Role of Circulating Unconjugated Estradiol in Mediating the Body Mass Index-Breast Cancer Association.
Higher body mass index (BMI) and circulating estrogen levels each increase postmenopausal breast cancer risk, particularly estrogen receptor-positive (ER(+)) tumors. Higher BMI also increases estrogen production. We estimated the proportion of the BMI-ER(+) breast cancer association mediated through estrogen in a case-control study nested within the Prostate, Lung, Colorectal, and Ovarian Cancer Screening Trial. Participants included 143 women with invasive ER(+) breast cancer and 268 matched controls, all postmenopausal and never having used hormone therapy at baseline. We used liquid chromatography-tandem mass spectrometry to measure 15 estrogens and estrogen metabolites in baseline serum. We calculated BMI from self-reported height and weight at baseline. We estimated the mediating effect of unconjugated estradiol on the BMI-ER(+) breast cancer association using Aalen additive hazards and Cox regression models. All estrogens and estrogen metabolites were statistically significantly correlated with BMI, with unconjugated estradiol most strongly correlated [Pearson correlation (r) = 0.45]. Approximately 7% to 10% of the effect of overweight, 12% to 15% of the effect of obesity, and 19% to 20% of the effect of a 5 kg/m(2) BMI increase on ER(+) breast cancer risk was mediated through unconjugated estradiol. The BMI-breast cancer association, once adjusted for unconjugated estradiol, was not modified by further adjustment for two metabolic ratios statistically significantly associated with both breast cancer and BMI. Circulating unconjugated estradiol levels partially mediate the BMI-breast cancer association, but other potentially important estrogen mediators (e.g., bioavailable estradiol) were not evaluated. Further research is required to identify mechanisms underlying the BMI-breast cancer association.


A monitoring system to provide feedback on student physical activity during physical education lessons.
School-based physical education (PE) provides opportunities to accumulate moderate-to-vigorous physical activity (MVPA), but many students are insufficiently active during PE lessons. Providing teachers with feedback regarding their students' physical activity may increase the effectiveness of PE for achieving MVPA goals, but existing physical activity monitoring technologies have limitations in class environments. Therefore, the purpose of this study was to develop and validate a system capable of providing feedback on PE lesson MVPA. Equations for translating step counts to %MVPA were derived from measures in 492 students who concurrently wore an ActiGraph GT3X+ (ActiGraph) and Yamax pedometer (Yamax) during a PE lesson. To enhance feedback availability during PE lessons, we then developed a bespoke monitoring system using wireless tri-axial pedometers (HMM) and a smart device app. After developing and testing the monitoring system, we assessed its validity and reliability in 100 students during a PE lesson. There was a strong correlation of 0.896 between step counts and accelerometer-determined %MVPA and quantile regression equations showed good validity for translating step counts to %MVPA with a mean absolute difference of 5.3 (95% CI, 4.4-6.2). The physical activity monitoring system was effective at providing %MVPA during PE lessons with a mean difference of 1.6 ± 7.1 compared with accelerometer-determined %MVPA (7% difference between the two measurement methods). Teachers and students can use a smart device app and wireless pedometers to conveniently obtain feedback during PE lessons. Future studies should determine whether such technologies help teachers to increase physical activity during PE lessons.


Simulation methods to estimate design power: an overview for applied research.
Estimating the required sample size and statistical power for a study is an integral part of study design. For standard designs, power equations provide an efficient solution to the problem, but they are unavailable for many complex study designs that arise in practice. For such complex study designs, computer simulation is a useful alternative for estimating study power. Although this approach is well known among statisticians, in our experience many epidemiologists and social scientists are unfamiliar with the technique. This article aims to address this knowledge gap. We review an approach to estimate study power for individual- or cluster-randomized designs using computer simulation. This flexible approach arises naturally from the model used to derive conventional power equations, but extends those methods to accommodate arbitrarily complex designs. The method is universally applicable to a broad range of designs and outcomes, and we present the material in a way that is approachable for quantitative, applied researchers. We illustrate the method using two examples (one simple, one complex) based on sanitation and nutritional interventions to improve child growth. We first show how simulation reproduces conventional power estimates for simple randomized designs over a broad range of sample scenarios to familiarize the reader with the approach. We then demonstrate how to extend the simulation approach to more complex designs. Finally, we discuss extensions to the examples in the article, and provide computer code to efficiently run the example simulations in both R and Stata. Simulation methods offer a flexible option to estimate statistical power for standard and non-traditional study designs and parameters of interest. The approach we have described is universally applicable for evaluating study designs used in epidemiologic and social science research.


Real-time neurofeedback using functional MRI could improve down-regulation of amygdala activity during emotional stimulation: a proof-of-concept study.
The amygdala is a central target of emotion regulation. It is overactive and dysregulated in affective and anxiety disorders and amygdala activity normalizes with successful therapy of the symptoms. However, a considerable percentage of patients do not reach remission within acceptable duration of treatment. The amygdala could therefore represent a promising target for real-time functional magnetic resonance imaging (rtfMRI) neurofeedback. rtfMRI neurofeedback directly improves the voluntary regulation of localized brain activity. At present, most rtfMRI neurofeedback studies have trained participants to increase activity of a target, i.e. up-regulation. However, in the case of the amygdala, down-regulation is supposedly more clinically relevant. Therefore, we developed a task that trained participants to down-regulate activity of the right amygdala while being confronted with amygdala stimulation, i.e. negative emotional faces. The activity in the functionally-defined region was used as online visual feedback in six healthy subjects instructed to minimize this signal using reality checking as emotion regulation strategy. Over a period of four training sessions, participants significantly increased down-regulation of the right amygdala compared to a passive viewing condition to control for habilitation effects. This result supports the concept of using rtfMRI neurofeedback training to control brain activity during relevant stimulation, specifically in the case of emotion, and has implications towards clinical treatment of emotional disorders.


Seasonal influenza vaccine coverage among high-risk populations in Thailand, 2010-2012.
The Advisory Committee on Immunization Practice of Thailand prioritizes seasonal influenza vaccinations for populations who are at highest risk for serious complications (pregnant women, children 6 months-2 years, persons ≥65 years, persons with chronic diseases, obese persons), and healthcare personnel and poultry cullers. The Thailand government purchases seasonal influenza vaccine for these groups. We assessed vaccination coverage among high-risk groups in Thailand from 2010 to 2012. National records on persons who received publicly purchased vaccines from 2010 to 2012 were analyzed by high-risk category. Denominator data from multiple sources were compared to calculate coverage. Vaccine coverage was defined as the proportion of individuals in each category who received the vaccine. Vaccine wastage was defined as the proportion of publicly purchased vaccines that were not used. From 2010 to 2012, 8.18 million influenza vaccines were publicly purchased (range, 2.37-3.29 million doses/year), and vaccine purchases increased 39% over these years. Vaccine wastage was 9.5%. Approximately 5.7 million (77%) vaccine doses were administered to persons ≥65 years and persons with chronic diseases, 1.4 million (19%) to healthcare personnel/poultry cullers, 82,570 (1.1%) to children 6 months-2 years, 78,885 (1.1%) to obese persons, 26,481 (0.4%) to mentally disabled persons, and 17,787 (0.2%) to pregnant women. Between 2010 and 2012, coverage increased among persons with chronic diseases (8.6% versus 14%; p<0.01) and persons ≥65 years (12%, versus 20%; p<0.01); however, coverage decreased for mentally disabled persons (6.1% versus 4.9%; p<0.01), children 6 months-2 years (2.3% versus 0.9%; p<0.01), pregnant women (1.1% versus 0.9%; p<0.01), and obese persons (0.2% versus 0.1%; p<0.01). From 2010 to 2012, the availability of publicly purchased vaccines increased. While coverage remained low for all target groups, coverage was highest among persons ≥65 years and persons with chronic diseases. Annual coverage assessments are necessary to promote higher coverage among high-risk groups in Thailand.


Association between energy drink intake, sleep, stress, and suicidality in Korean adolescents: energy drink use in isolation or in combination with junk food consumption.
A considerable amount of research suggests that the frequent use of caffeinated energy drinks may be associated with undesirable effects, particularly so in children and adolescents. This study aimed to investigate the associations between energy drink intake and mental health problems, in isolation or in combination with junk food consumption, in a nationally representative sample of Korean adolescents. Data from the 2015 Korean Youth Risk Behavior Web-Based Survey, collected from 68,043 adolescents aged 12-18 years (mean age 15.09 ± 1.72 years), were analyzed. Questionnaires were administered to collect information related to dietary behavior including energy drink intake and junk food consumption. Single item measures of sleep dissatisfaction, stress, depression, suicidal ideation, suicide plan, and suicide attempt were also administered. Associations between energy drink intake and sleep dissatisfaction, perceived severe stress, persistent depressive mood, and suicidality were investigated, and a multivariate approach was taken so that additional variance from demographic and lifestyle factors could be controlled for statistically. Energy drink intake was significantly associated with sleep dissatisfaction (adjusted odd ratios [AORs] = 1.64 and 1.25), severe stress (AORs = 2.23 and 1.38), depressive mood (AOR = 2.59 and 1.51), suicidal ideation (AORs = 3.14 and 1.43), suicide plan (AORs = 4.65 and 1.78), and suicide attempt (AORs = 6.79 and 1.91), with a higher risk for more frequent use of energy drinks (≥5 times/wk) than for less frequent use (1-4 times/wk). The detrimental effect of energy drinks on mental health was particularly prominent in frequent junk food consumers. Our data suggest that energy drink intake had detrimental effects related to stress, sleep dissatisfaction, mood, and suicidality, in isolation or in combination with junk food consumption, in Korean adolescents. However, the cross-sectional study design prevents our ability to assess causal relationships.


Umbilical arterial pH < 7.00 in newborns delivered by nonelective cesarean delivery: risk factors and peripartum outcomes.
The objective of this study was to compare the risk factors and peripartum outcomes among patients who had nonelective cesarean delivery with umbilical arterial pH < versus > or = 7.00. A case-control study of parturients who delivered by nonelective cesarean delivery and had a neonate with a pH < 7.00 were compared with the next four patients who delivered abdominally but had a newborn with a normal pH. Among 45 newborns with pH < 7.00 the rate of cesarean delivery for nonreassuring fetal heart rate was significantly more common in the case (56%) than control group (16%). The rates of end organ failure and neonatal death were similar for both groups. Although newborns with pH < 7.00 were significantly more likely to have cesarean delivery for nonreassuring fetal heart rate patterns and be admitted to the neonatal intensive care unit, there was no means to identify these patients until the abnormalities in tracing developed.


Defects in pancreatic development and glucose metabolism in SMN-depleted mice independent of canonical spinal muscular atrophy neuromuscular pathology.
Spinal muscular atrophy (SMA) is characterized by motor neuron loss, caused by mutations or deletions in the ubiquitously expressed survival motor neuron 1 (SMN1) gene. We recently identified a novel role for Smn protein in glucose metabolism and pancreatic development in both an intermediate SMA mouse model (Smn(2B/-)) and type I SMA patients. In the present study, we sought to determine if the observed metabolic and pancreatic defects are SMA-dependent. We employed a line of heterozygous Smn-depleted mice (Smn(+/-)) that lack the hallmark SMA neuromuscular pathology and overt phenotype. At 1 month of age, pancreatic/metabolic function of Smn(+/-)mice is indistinguishable from wild type. However, when metabolically challenged with a high-fat diet, Smn(+/-)mice display abnormal localization of glucagon-producing α-cells within the pancreatic islets and increased hepatic insulin and glucagon sensitivity, through increased p-AKT and p-CREB, respectively. Further, aging results in weight gain, an increased number of insulin-producing β cells, hyperinsulinemia and increased hepatic glucagon sensitivity in Smn(+/-)mice. Our study uncovers and highlights an important function of Smn protein in pancreatic islet development and glucose metabolism, independent of canonical SMA pathology. These findings suggest that carriers of SMN1 mutations and/or deletions may be at an increased risk of developing pancreatic and glucose metabolism defects, as even small depletions in Smn protein may be a risk factor for diet- and age-dependent development of metabolic disorders.


Estimation of the secretion rate of insulin from the urinary excretion rate of C-peptide. Study in obese and diabetic subjects.
Direct methods for measuring the secretion rate of insulin are too cumbersome for clinical application. Since C-peptide is secreted in an equimolar ratio with insulin and is excreted into the urine, measuring the urinary excretion rate of C-peptide (U-C) could serve as an indicator of its secretion rate (SR-C) if its urinary clearance (UCI-C) is constant and unaffected by plasma C-peptide concentration, body mass, or diabetes. We measured clearance ratios of C-peptide/creatinine (CR) in the fasting state and integrated 0-1, 1-3, and 3-5 h after 100 g of glucose p.o. as well as over a full 24-h in eight obese, eight lean, and six maturity-onset diabetic subjects. CR did not differ significantly when values in the fasting state were compared with those in the postprandial periods and was therefore unaffected by plasma C-peptide concentration. Furthermore, CR was similar in the lean, obese, and diabetic subjects. SR-C, determined as the product of the metabolic clearance rate of C-peptide and its fasting or integrated plasma concentrations, correlated significantly with U-C in all the subjects (r = 0.87, P less than 0.0001). The correlation of U-C with SR-C in the diabetic subjects alone was also significant (r = 0.88, P less than 0.0001). In conclusion, our data support the use of U-C as an indirect measure of SR-C and therefore of SR-I.


Differential distribution of Langerhans cells in organ culture of human skin.
Epidermal Langerhans cells (LCs) function as the antigen-presenting cells in such cutaneous cell-mediated immune responses as contact hypersensitivity and in the mixed epidermal cell-lymphocyte reaction. They have also been implicated in the immune response in skin allograft rejection. Since organ culture of thyroid and pancreas has been shown to prolong allograft survival, presumably through the loss of antigen-presenting cells, we examined the effect of skin explant culture on LC survival. Human skin explants were placed in organ culture and examined serially as whole mounts of epidermis for the presence of LCs as judged by ATPase activity, and OKT-6 and HLA-DR antigens. Although we observed morphologic changes and an absolute reduction in the number of positively stained cells, culture for up to 28 days failed to deplete explants of these cells. Langerhans cells were also sought in the epidermal outgrowths that develop peripheral to the original explants. They were never seen in the area beyond 0.3 mm from the explant edge. Organ culture of skin thus provides a means to explore the contribution of LCs to skin allograft rejection by comparing the immunogenicity of epidermal portions of the explant with the epidermal outgrowth.


iBarcode.org: web-based molecular biodiversity analysis.
DNA sequences have become a primary source of information in biodiversity analysis. For example, short standardized species-specific genomic regions, DNA barcodes, are being used as a global standard for species identification and biodiversity studies. Most DNA barcodes are being generated by laboratories that have an expertise in DNA sequencing but not in bioinformatics data analysis. Therefore, we have developed a web-based suite of tools to help the DNA barcode researchers analyze their vast datasets. Our web-based tools, available at http://www.ibarcode.org, allow the user to manage their barcode datasets, cull out non-unique sequences, identify haplotypes within a species, and examine the within- to between-species divergences. In addition, we provide a number of phylogenetics tools that will allow the user to manipulate phylogenetic trees generated by other popular programs. The use of a web-based portal for barcode analysis is convenient, especially since the WWW is inherently platform-neutral. Indeed, we have even taken care to ensure that our website is usable from handheld devices such as PDAs and smartphones. Although the current set of tools available at iBarcode.org were developed to meet our own analytic needs, we hope that feedback from users will spark the development of future tools. We also welcome user-built modules that can be incorporated into the iBarcode framework.


Pleurectomy and decortication.
Pleurectomy and decortication (P/D) improve survival and quality of life in selected patients with malignant pleural mesothelioma. The operative procedure was not standardized until recently. The goal of the operation is to perform a macroscopic complete resection of the tumor. This often involves resection of the parietal and visceral pleura and invariably a partial or complete resection and prosthetic reconstruction of ipsilateral hemidiaphragm. We describe our operative planning and technique as well as outcomes of P/D reported in current literature.


Chronic exposure to cigarette smoke condensate in vitro induces epithelial to mesenchymal transition-like changes in human bronchial epithelial cells, BEAS-2B.
Cigarette smoke causes lung tumorigenesis; however, the mechanisms underlying transformation are unknown. We investigated if tobacco compounds induce DNA promoter hypermethylation in BEAS-2B cells treated with low doses of cigarette smoke condensate (CSC) for one month. Transcriptional profiles and anchorage-independent growth were explored using Affymetrix microarray and soft agar assay, respectively. To investigate if tobacco compounds induce hypermethylation, CSC/dimethyl sulfoxide (DMSO)-treated cells were further treated with 5-Aza-2'-deoxycytidine (5AzaC) and trychostatin A (TSA). This treatment was followed by transcriptional profiling. CSC-exposed cells acquired a fibroblast-like shape with enhanced anchorage-independent growth. Silencing of epithelial cadherin, the hallmark of epithelial to mesenchymal transition (EMT), was observed upon exposure to CSC. Changes in the expression of genes involved in epidermal development, intercellular junction formation, and cytoskeleton formation were identified. Gene expression profiles from 5AzaC- and TSA-treated cells revealed 130 genes possibly methylated due to chronic CSC exposure. Our results suggest that E-cadherin may also be silenced by hypermethylation in an in vitro model of chronic exposure to low doses of CSC. This study demonstrates evidence for a tobacco compound induced EMT-like process in vitro and provides insight into possible mechanisms of gene silencing occurring during this treatment.


Non-erosive reflux disease (NERD) and erosive esophagitis--a spectrum of disease or special entities?
An understanding of the natural history of GERD has an important impact on the long-term management of the disorder. By assessing the currently available studies that evaluated the natural course of GERD, a new conceptual framework that suggests that GERD is composed of three distinct phenotypic presentations has been recently proposed. Presently, the field has been divided into two camps that support or oppose a paradigm shift in the natural course of GERD. Nevertheless, the largest population-based or longest duration natural course studies report only 10% progress from NERD to erosive esophagitis over time. The other GERD patients remained within their respective phenotypic presentation of GERD.


Abdominal Contouring: Can the American Society of Anesthesiologists Classification System Help Determine When to Say No?
The purpose of this study was to determine whether the American Society of Anesthesiologists classification system could be used preoperatively to identify patients at high risk for complications after abdominal contouring. Using the National Surgical Quality Improvement Program database from 2007 to 2012, patients undergoing abdominal contouring procedures were identified and stratified by American Society of Anesthesiologists class. The primary outcome was any complication within 30 days. Secondary outcomes included minor wound, major surgical, and medical complication. Adjusted odds ratios were calculated using logistic regression. A total of 3637 patients were analyzed; 14.6 percent of patients were class I, 59.1 percent were class II, 23.4 percent were class III, and 2.9 percent were class IV. Overall complication and mortality rates were 12 percent and 0.2 percent, respectively. There was a significant trend of increasing odds of any complication with increasing class (class I, OR, 1.0; class II, OR, 1.5; class III, OR, 2.5; class IV, OR, 5.6; p-trend < 0.001). This trend was seen consistently for minor wound complications, medical complications, and major surgical complications (p = 0.007, p = 0.005, and p = 0.001, respectively). The American Society of Anesthesiologists classification system, which is simple and universally applicable, appears to predict significant complications and can be used to rapidly screen patients before abdominal contouring. Furthermore, the authors' results can be used to inform patient-physician discussion about the risks incurred when undergoing these procedures based on their individual class. Together with optimization of high-risk patients, patient selection using American Society of Anesthesiologists classification may prevent complications and improve outcomes. Risk, II.


Cloning and expression of a rat brain GABA transporter.
A complementary DNA clone (designated GAT-1) encoding a transporter for the neurotransmitter gamma-aminobutyric acid (GABA) has been isolated from rat brain, and its functional properties have been examined in Xenopus oocytes. Oocytes injected with GAT-1 synthetic messenger RNA accumulated [3H]GABA to levels above control values. The transporter encoded by GAT-1 has a high affinity for GABA, is sodium-and chloride-dependent, and is pharmacologically similar to neuronal GABA transporters. The GAT-1 protein shares antigenic determinants with a native rat brain GABA transporter. The nucleotide sequence of GAT-1 predicts a protein of 599 amino acids with a molecular weight of 67 kilodaltons. Hydropathy analysis of the deduced protein suggests multiple transmembrane regions, a feature shared by several cloned transporters; however, database searches indicate that GAT-1 is not homologous to any previously identified proteins. Therefore, GAT-1 appears to be a member of a previously uncharacterized family of transport molecules.


Enfuvirtide antiretroviral therapy in HIV-1 infection.
It has been over 25 years since the first diagnosis of what would be known as AIDS. Although great strides in anti-HIV therapeutics have been made, there is still a great need for antiretrovirals that are effective against drug-resistant HIV. Enfuvirtide (ENF) is the first of a new class of fusion inhibitors to be approved by the US Food and Drug Administration for use in combination with other antiretroviral agents among HIV-1 infected patients with previous treatment experience. The inclusion of enfuvirtide in an optimized antiretroviral background regimen for the treatment of HIV-1 infected (treatment-experienced) patients followed the success of two critical clinical trials (TORO: T20 vs Optimized Regimen Only I and II). Even though injection-site reactions persisted in these trials, improved virological and immunological responses were observed among patients. Challenges associated with ENF treatment include the high cost of the drug, injection-site reactions, determining the optimal time to initiate treatment, and the potential for the selection of drug resistant mutants and viral evolution. ENF is a promising novel treatment for HIV infected individuals whose choices for effective treatment are limited by previous treatment and resistance. Understanding the implications of viral fitness and evolution in the presence of ENF treatment is crucial in determining effective and safe treatment regimens, particularly among treatment-experienced patients.


Pre- and postnatal Lactobacillus reuteri treatment alters DNA methylation of infant T helper cells.
Perinatal childhood exposures, including probiotic supplementation, may affect epigenetic modifications and impact on immune maturation and allergy development. The aim of this study was to assess the effects of pre- and postnatal Lactobacillus reuteri supplementation on DNA methylation in relation to immune maturation and allergy development. DNA methylation patterns were investigated for allergy-related T helper subsets using a locus-specific method and at a genome-wide scale using the Illumina 450K array. From a randomised, double-blind, placebo-controlled allergy prevention trial with pre- and postnatal probiotic supplementation, CD4+ T helper cells were obtained at birth (from cord blood), and 12 and 24 months of age (total (placebo/probiotics); locus-specific method: CB = 32 (17/15), 12 months = 24 (9/15), 24 months = 35 (15/20); Illumina: CB = 19 (10/9), 12 months = 10 (6/4), 24 months = 19(11/8)). Comparing probiotics to placebo, the greatest genome-wide differential DNA methylation was observed at birth, where the majority of sites were hypomethylated, indicating transcriptional accessibility in the probiotic group. Bioinformatic analyses, including network analyses, revealed a module containing 91 genes, enriched for immune-related pathways such as chemotaxis, PI3K-Akt, MAPK and TGF-β signalling. A majority of the module genes were associated with atopic manifestations (OR = 1.43, P = 2.4 × 10-6 ), and a classifier built on this model could predict allergy development (AUC = 0.78, P = 3.0 × 10e-3 ). Pathways such as IFN-γ signalling and T-cell activation were more hypermethylated at birth compared with later in life in both intervention groups over time, in line with DNA methylation patterns in the IFNG locus obtained by the locus-specific methodology. Maternal L. reuteri supplementation during pregnancy alters DNA methylation patterns in CD4+ T cells towards enhanced immune activation at birth, which may affect immune maturation and allergy development.


Effects of dietary polyvinylchloride microparticles on general health, immune status and expression of several genes related to stress in gilthead seabream (Sparus aurata L.).
It is a long-recognized fact that marine plastic debris contaminates the oceans and seas of the entire world. Even though their effects on the aquatic biota are not well documented or understood. The effects of dietary polyvinylchloride microparticles (PVC-MPs) on the general health, immune status and some stress markers were studied using gilthead seabream (Sparus aurata) as a model of marine fish. Thirty specimens were randomly placed in three running sea water aquaria and fish in each aquarium received an experimental diet containing 0 (control), 100 or 500 mg kg-1 of PVC-MPs for 30 days. Metabolic parameters in serum indicated that the dietary intake of PVC-MPs negatively affected several vital organs. Humoral immune parameters were determined in serum and skin mucus. Cellular immune parameters were determined in head-kidney leucocytes. Concomitantly, the expression of different genes related to stress was studied in head-kidney and liver. Regarding head-kidney gene expression, prdx5 was significantly decreased by PVC-MPs intake for 15 and 30 days, respect to the values found in control fish. On the other hand, the expression of prdx1 and prdx3 were significantly increased by the PVC-MPs intake during 15 and 30 days, compared with the values found in control fish. Furthermore, the expression of hsp90 and ucp1 genes decreased and increased, respectively, in the liver of fish fed 500 mg kg-1 of PVC-MPs for 30 days. Although ingestion of PVC-MPs provoked few significant effects (mostly increases) in the main immune activities of gilthead seabream compared with the values found in control fish, PVC-MPs are recognized by the fish as stressors. Continued exposure of fish to high concentrations of PVC-MPs could have a negative impact on fish physiology due to the chronic stress produced.


The Role of Anharmonicity in Hydrogen-Bonded Systems: The Case of Water Clusters.
The nature of vibrational anharmonicity has been examined for the case of small water clusters using second-order vibrational perturbation theory (VPT2) applied on second-order Møller-Plesset perturbation theory (MP2) potential energy surfaces. Using a training set of 16 water clusters (H2O)n=2-6,8,9 with a total of 723 vibrational modes, we determined scaling factors that map the harmonic frequencies onto anharmonic ones. The intermolecular modes were found to be substantially more anharmonic than intramolecular bending and stretching modes. Due to the varying levels of anharmonicity of the intermolecular and intramolecular modes, different frequency scaling factors for each region were necessary to achieve the highest accuracy. Furthermore, new scaling factors for zero-point vibrational energies (ZPVE) and vibrational corrections to the enthalpy (ΔHvib) and the entropy (Svib) have been determined. All the scaling factors reported in this study are different from previous works in that they are intended for hydrogen-bonded systems, while others were built using experimental frequencies of covalently bonded systems. An application of our scaling factors to the vibrational frequencies of water dimer and thermodynamic functions of 11 larger water clusters highlights the importance of anharmonic effects in hydrogen-bonded systems.


Expedited liver allocation in the United States: a critical analysis.
The fate of donor livers allocated via an out-of-sequence expedited placement (EP) pathway has not been previously examined. We determined the originating and receiving United Network for Organ Sharing (UNOS) regions of all donor livers procured between January 1, 2010 and October 31, 2012 and placed out of sequence with UNOS bypass code 863 (EP attempt) or 898 (miscellaneous). We reviewed the early function of these liver grafts and assessed the effect of EP allocation on wait-listed patients at our center. Registrants at our center were eligible to receive 1298 liver offers during the interval studied: 218 (16.8%) of these liver offers bypassed our center and were allocated to other centers and used in patients lower on the match-run list. During the study interval, 560 livers were allocated in the United States by EP. Regions 1, 5, 7, 9, and 10 used the greatest number of EP-placed grafts. Region 1 (New England) used the greatest proportion of all EP livers (33% of all imported EP livers in the United States, P < 0.001 versus all other regions). Graft function data were available for 560 livers placed by EP: 491 (88%) of these grafts were functioning at a mean of 399.5 days after transplantation. In conclusion, the transplantation of livers allocated by means of an expedited refusal code is asymmetric across regions and, in some instances, results in the bypassing of patients with higher wait-list priority but without notification of the bypassed center. Short-term graft function after EP allocation is excellent. Policies governing EP allocation should be created in order to improve access to available organs.


Impaired sucrose induction1 encodes a conserved plant-specific protein that couples carbohydrate availability to gene expression and plant growth.
To identify the molecular mechanisms underlying carbohydrate allocation to storage processes, we have isolated mutants in which the sugar induction of starch biosynthetic gene expression was impaired. Here we describe the IMPAIRED SUCROSE INDUCTION1 (ISI1) gene, which encodes a highly conserved plant-specific protein with structural similarities to Arm repeat proteins. ISI1 is predominantly expressed in the phloem of leaves following the sink-to-source transition during leaf development, but is also sugar-inducible in mesophyll cells. Soil-grown isi1 mutants show reduced plant growth and seed set compared to wild-type Arabidopsis. This growth reduction is not due to reduced carbohydrate availability or a defect in sucrose export from mature leaves, suggesting that isi1 mutant plants do not utilize available carbohydrate resources efficiently. ISI1 interacts synergistically with, but is genetically distinct from, the abscisic acid (ABA) signalling pathway controlling sugar responses via ABI4. Our data show that ISI1 couples the availability of carbohydrates to the control of sugar-responsive gene expression and plant growth.


Four-dimensional visualization of subpleural alveolar dynamics in vivo during uninterrupted mechanical ventilation of living swine.
Pulmonary alveoli have been studied for many years, yet no unifying hypothesis exists for their dynamic mechanics during respiration due to their miniature size (100-300 μm dimater in humans) and constant motion, which prevent standard imaging techniques from visualizing four-dimensional dynamics of individual alveoli in vivo. Here we report a new platform to image the first layer of air-filled subpleural alveoli through the use of a lightweight optical frequency domain imaging (OFDI) probe that can be placed upon the pleura to move with the lung over the complete range of respiratory motion. This device enables in-vivo acquisition of four-dimensional microscopic images of alveolar airspaces (alveoli and ducts), within the same field of view, during continuous ventilation without restricting the motion or modifying the structure of the alveoli. Results from an exploratory study including three live swine suggest that subpleural alveolar air spaces are best fit with a uniform expansion (r (2) = 0.98) over a recruitment model (r (2) = 0.72). Simultaneously, however, the percentage change in volume shows heterogeneous alveolar expansion within just a 1 mm x 1 mm field of view. These results signify the importance of four-dimensional imaging tools, such as the device presented here. Quantification of the dynamic response of the lung during ventilation may help create more accurate modeling techniques and move toward a more complete understanding of alveolar mechanics.


Lip Reconstruction after Tumor Ablation.
Approximately 25% of all oral cavity carcinomas involve the lips, and the primary management of these lesions is complete surgical resection. Loss of tissue in the lips after resection is treated with a variety of techniques, depending on the extension and location of the defect. Here we review highly accepted techniques of lip reconstruction and some of new trials with significant clinical results. Reconstruction choice is primarily depend to size of the defect, localization of defect, elasticity of tissues. But patient's age, comorbidities, and motivation are also important. According to the defect location and size, different reconstruction methods can be used. For defects involved less than 30% of lips, primary closures are sufficient. In defects with 35-70% lip involvement, the Karapandzic, Abbe, Estlander, McGregor or Gillies' fan flaps or their modifications can be used. When lip remaining tissues are insufficient, cheek tissue can be used in Webster and Bernard advancement flaps and their various modifications. Deltopectoral or radial forearm free flaps can be options for large defects of the lip extending to the Jaws. To achieve best functional and esthetic results, surgeons should be able to choose most appropriate reconstruction method. Considering defects' size and location, patients' expects and surgeon's ability and knowledge, a variety of flaps are presented in order to reconstruct defects resulted from tumor ablation. It's necessary for surgeons to trace the recent innovations in lip reconstruction to offer best choices to patients.


[Global Allergy and Asthma European Network (GA2LEN) addresses the epidemic of allergy and asthma].
Allergic diseases represent a major health problem in Europe. They are increasing in prevalence, severity and costs. GA2LEN, an FP6 Network of Excellence, was created in 2005 as a vehicle to ensure excellence in research bringing together research and clinical institutions to combat fragmentation in the European research area and to tackle Allergy in its globality. GA2LEN benefited greatly from the voluntary efforts of researchers who are strongly committed to this model of pan-European collaboration. The network was organized in order to increase networking for scientific projects in allergy and asthma around Europe and to make GA2LEN the world leader in the field. Besides these activities, research has been jointly made and the first papers are being published. GA2LEN achievements in general can be grouped as those for a durable infrastructure built up during the project phase those which are project-related work based on these novel infrastructures, and the development and implementations of guidelines. The major achievements of GA2LEN are reported in this paper.


A heterozygous deletion mutation in the cardiac sodium channel gene SCN5A with loss- and gain-of-function characteristics manifests as isolated conduction disease, without signs of Brugada or long QT syndrome.
The SCN5A gene encodes for the α-subunit of the cardiac sodium channel NaV1.5, which is responsible for the rapid upstroke of the cardiac action potential. Mutations in this gene may lead to multiple life-threatening disorders of cardiac rhythm or are linked to structural cardiac defects. Here, we characterized a large family with a mutation in SCN5A presenting with an atrioventricular conduction disease and absence of Brugada syndrome. In a large family with a high incidence of sudden cardiac deaths, a heterozygous SCN5A mutation (p.1493delK) with an autosomal dominant inheritance has been identified. Mutation carriers were devoid of any cardiac structural changes. Typical ECG findings were an increased P-wave duration, an AV-block I° and a prolonged QRS duration with an intraventricular conduction delay and no signs for Brugada syndrome. HEK293 cells transfected with 1493delK showed strongly (5-fold) reduced Na(+) currents with altered inactivation kinetics compared to wild-type channels. Immunocytochemical staining demonstrated strongly decreased expression of SCN5A 1493delK in the sarcolemma consistent with an intracellular trafficking defect and thereby a loss-of-function. In addition, SCN5A 1493delK channels that reached cell membrane showed gain-of-function aspects (slowing of the fast inactivation, reduction in the relative fraction of channels that fast inactivate, hastening of the recovery from inactivation). In a large family, congregation of a heterozygous SCN5A gene mutation (p.1493delK) predisposes for conduction slowing without evidence for Brugada syndrome due to a predominantly trafficking defect that reduces Na(+) current and depolarization force.


Detection of 3D atomic similarities and their use in the discrimination of small molecule protein-binding sites.
Current computational methods for the prediction of function from structure are restricted to the detection of similarities and subsequent transfer of functional annotation. In a significant minority of cases, global sequence or structural (fold) similarities do not provide clues about protein function. In these cases, one alternative is to detect local binding site similarities. These may still reflect more distant evolutionary relationships as well as unique physico-chemical constraints necessary for binding similar ligands, thus helping pinpoint the function. In the present work, we ask the following question: is it possible to discriminate within a dataset of non-homologous proteins those that bind similar ligands based on their binding site similarities? We implement a graph-matching-based method for the detection of 3D atomic similarities introducing some simplifications that allow us to extend its applicability to the analysis of large allatom binding site models. This method, called IsoCleft, does not require atoms to be connected either in sequence or space. We apply the method to a cognate-ligand bound dataset of non-homologous proteins. We define a family of binding site models with decreasing knowledge about the identity of the ligand-interacting atoms to uncouple the questions of predicting the location of the binding site and detecting binding site similarities. Furthermore, we calculate the individual contributions of binding site size, chemical composition and geometry to prediction performance. We find that it is possible to discriminate between different ligand-binding sites. In other words, there is a certain uniqueness in the set of atoms that are in contact to specific ligand scaffolds. This uniqueness is restricted to the atoms in close proximity of the ligand in which case, size and chemical composition alone are sufficient to discriminate binding sites. Discrimination ability decreases with decreasing knowledge about the identity of the ligand-interacting binding site atoms. The decrease is quite abrupt when considering size and chemical composition alone, but much slower when including geometry. We also observe that certain ligands are easier to discriminate. Interestingly, the subset of binding site atoms belonging to highly conserved residues is not sufficient to discriminate binding sites, implying that convergently evolved binding sites arrived at dissimilar solutions. IsoCleft can be obtained from the authors.


The top 10 research priorities in diabetes and pregnancy according to women, support networks and healthcare professionals.
To undertake a Priority Setting Partnership (PSP) to establish priorities for future research in diabetes and pregnancy, according to women with experience of pregnancy, and planning pregnancy, with any type of diabetes, their support networks and healthcare professionals. The PSP used established James Lind Alliance (JLA) methodology working with women and their support networks and healthcare professionals UK-wide. Unanswered questions about the time before, during or after pregnancy with any type of diabetes were identified using an online survey and broad-level literature search. A second survey identified a shortlist of questions for final prioritisation at an online consensus development workshop. There were 466 responses (32% healthcare professionals) to the initial survey, with 1161 questions, which were aggregated into 60 unanswered questions. There were 614 responses (20% healthcare professionals) to the second survey and 18 questions shortlisted for ranking at the workshop. The top 10 questions were: diabetes technology, the best test for diabetes during pregnancy, diet and lifestyle interventions for diabetes management during pregnancy, emotional and well-being needs of women with diabetes pre- to post-pregnancy, safe full-term birth, post-natal care and support needs of women, diagnosis and management late in pregnancy, prevention of other types of diabetes in women with gestational diabetes, women's labour and birth experiences and choices and improving planning pregnancy. These research priorities provide guidance for research funders and researchers to target research in diabetes and pregnancy that will achieve greatest value and impact.


Trifurcate feed-forward regulation of age-dependent cell death involving miR164 in Arabidopsis.
Aging induces gradual yet massive cell death in higher organisms, including annual plants. Even so, the underlying regulatory mechanisms are barely known, despite the long-standing interest in this topic. Here, we demonstrate that ORE1, which is a NAC (NAM, ATAF, and CUC) transcription factor, positively regulates aging-induced cell death in Arabidopsis leaves. ORE1 expression is up-regulated concurrently with leaf aging by EIN2 but is negatively regulated by miR164. miR164 expression gradually decreases with aging through negative regulation by EIN2, which leads to the elaborate up-regulation of ORE1 expression. However, EIN2 still contributes to aging-induced cell death in the absence of ORE1. The trifurcate feed-forward pathway involving ORE1, miR164, and EIN2 provides a highly robust regulation to ensure that aging induces cell death in Arabidopsis leaves.


New technique for early screening of developmental dysplasia of the hip: pilot study.
An acoustical technique has been developed for early screening of developmental dysplasia of the hip (DDH) in neonates by comparing the sound transmitted across the hips while a vibratory force was applied to the sacrum. The baseline for 90 normal neonates has been established and tests in the frequency bands of 200, 250, and 315 Hz were found to be most effective, achieving high coherence and smallest discrepancy. Sixteen patients with unilateral DDH were examined and the results suggested that coherence below 0.8 in at least one of these frequency bands was strongly indicative of structural asymmetry between both hips, and there was a significant difference between normal neonates and patients with unilateral DDH. By setting the cut-off discrepancy at 2.0 dB, the best sensitivity (100%) would be achieved, and this could be proposed as the threshold for wide-scale screening of DDH.


Functionally reversible impacts of disturbances on lake food webs linked to spatial and seasonal dependencies.
Increasing human impact on the environment is causing drastic changes in disturbance regimes and how they prevail over time. Of increasing relevance is to further our understanding on biological responses to pulse disturbances (short duration) and how they interact with other ongoing press disturbances (constantly present). Because the temporal and spatial contexts of single experiments often limit our ability to generalize results across space and time, we conducted a modularized mesocosm experiment replicated in space (five lakes along a latitudinal gradient in Scandinavia) and time (two seasons, spring and summer) to generate general predictions on how the functioning and composition of multitrophic plankton communities (zoo-, phyto- and bacterioplankton) respond to pulse disturbances acting either in isolation or combined with press disturbances. As pulse disturbance, we used short-term changes in fish presence, and as press disturbance, we addressed the ongoing reduction in light availability caused by increased cloudiness and lake browning in many boreal and subarctic lakes. First, our results show that the top-down pulse disturbance had the strongest effects on both functioning and composition of the three trophic levels across sites and seasons, with signs for interactive impacts with the bottom-up press disturbance on phytoplankton communities. Second, community composition responses to disturbances were highly divergent between lakes and seasons: temporal accumulated community turnover of the same trophic level either increased (destabilization) or decreased (stabilization) in response to the disturbances compared to control conditions. Third, we found functional recovery from the pulse disturbances to be frequent at the end of most experiments. In a broader context, these results demonstrate that top-down, pulse disturbances, either alone or with additional constant stress upon primary producers caused by bottom-up disturbances, can induce profound but often functionally reversible changes across multiple trophic levels, which are strongly linked to spatial and temporal context dependencies. Furthermore, the identified dichotomy of disturbance effects on the turnover in community composition demonstrates the potential of disturbances to either stabilize or destabilize biodiversity patterns over time across a wide range of environmental conditions.


NR2C in the thalamic reticular nucleus; effects of the NR2C knockout.
NMDAR antagonists can evoke delta frequency bursting in the nucleus reticularis of the thalamus (nRT). The mechanism of this oscillation was determined; antagonist blocks an NR2C-like conductance that has low Mg block at resting potential and thus can contribute a resting inward current in response to ambient glutamate. Block of this current hyperpolarizes the cell, deinactivating T-type Ca channels and thus triggering delta frequency bursting. The basis for assuming a NR2C-like conductance was that (1) transcripts for NR2C are abundant in the thalamus and (2) the current-voltage curve of the synaptically evoked NMDAR current has the low rectification characteristic of NR2C. In the current study, we have sought to determine whether the channels that generate the NMDAR current are NR2C-like or are actually comprised of receptors containing NR2C. We studied the current-voltage curve of synaptically evoked NMDAR current in the nRT of NR2C knockout mice. In wild-type mice, the current was weakly voltage dependent, as previously observed in rats. This weak rectification was absent in NR2C KO mice. In contrast, NR2C KO had no effect on the strongly rectifying NMDAR current in pyramidal cells of the prefrontal cortex. These results demonstrate that the low rectification normally observed in the nRT is due to NR2C.


Assessment of the efficacy of chelate-assisted phytoextraction of lead by coffeeweed (Sesbania exaltata Raf.).
Lead (Pb), depending upon the reactant surface, pH, redox potential and other factors can bind tightly to the soil with a retention time of many centuries. Soil-metal interactions by sorption, precipitation and complexation processes, and differences between plant species in metal uptake efficiency, transport, and susceptibility make a general prediction of soil metal bioavailability and risks of plant metal toxicity difficult. Moreover, the tight binding characteristic of Pb to soils and plant materials make a significant portion of Pb unavailable for uptake by plants. This experiment was conducted to determine whether the addition of ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), or acetic acid (HAc) can enhance the phytoextraction of Pb by making the Pb soluble and more bioavailable for uptake by coffeeweed (Sesbania exaltata Raf.). Also we wanted to assess the efficacy of chelates in facilitating translocation of the metal into the above-ground biomass of this plant. To test the effect of chelates on Pb solubility, 2 g of Pb-spiked soil (1000 mg Pb/kg dry soil) were added to each 15 mL centrifuge tube. Chelates (EDTA, EGTA, HAc) in a 1:1 ratio with the metal, or distilled deionized water were then added. Samples were shaken on a platform shaker then centrifuged at the end of several time periods. Supernatants were filtered with a 0.45 mum filter and quantified by inductively coupled plasma-optical emission spectrometry (ICP-OES) to determine soluble Pb concentrations. Results revealed that EDTA was the most effective in bringing Pb into solution, and that maximum solubility was reached 6 days after chelate amendment. Additionally, a greenhouse experiment was conducted by planting Sesbania seeds in plastic tubes containing top soil and peat (2:1, v:v) spiked with various levels (0, 1000, 2000 mg Pb/kg dry soil) of lead nitrate. At six weeks after emergence, aqueous solutions of EDTA and/or HAc (in a 1:1 ratio with the metal) or distilled deionized water were applied to the root zones. Plants were harvested at 6 days after chelate addition to coincide with the duration of maximum metal solubility previously determined in this study. Results of the greenhouse experiment showed that coffeeweed was relatively tolerant to moderate levels of Pb and chelates as shown by very slight reductions in root and no discernable effects on shoot biomass. Root Pb concentrations increased with increasing levels of soil-applied Pb. Further increases in root Pb concentrations were attributed to chelate amendments. In the absence of chelates, translocation of Pb from roots to shoots was minimal. However, translocation dramatically increased in treatments with EDTA alone or in combination with HAc. Overall, the results of this study indicated that depending on the nature and type of Pb-contaminated soil being remediated, the bioavailability and uptake of Pb by coffeeweed can be enhanced by amending the soil with chelates especially after the plants have reached maximum biomass.


Computed tomographic distinction of perirenal liposarcoma from exophytic angiomyolipoma: a feature analysis study.
To identify computed tomographic (CT) findings that distinguish perirenal liposarcomas from exophytic angiomyolipomas. We identified CT scans of 20 patients (13 women and 7 men; mean age, 63 years) with either perirenal liposarcoma (n = 11) proven at histopathology or large exophytic angiomyolipomas (n = 9) determined by 2-year stability (n = 6) or histopathology (n = 3). Two independent readers unaware of the final diagnoses recorded the presence of the following CT findings: (1) tumoral vessel extending into the renal cortex, (2) tumoral vessel extending into the renal hilum, (3) renal parenchymal defect at the site of tumor contact, (4) intratumoral hemorrhage, (5) nonfat attenuating intratumoral nodules, and (6) calcification. A tumoral vessel extending into the renal cortex was seen only in angiomyolipomas (7 and 6 of 9 patients versus 0 and 0 of 11 liposarcomas for readers 1 and 2, respectively; P < 0.005 for both). A parenchymal defect was more commonly seen in angiomyolipomas (7 and 6 of 9 angiomyolipomas versus 1 and 1 of 11 liposarcomas for readers 1 and 2, respectively; P < 0.05 for both). Calcifications were seen by both readers in 6 of 11 liposarcomas but not in any angiomyolipomas (P < 0.05). The other recorded findings were not useful in distinguishing CT features (P > 0.1 for both readers). In the evaluation of a fatty perinephric mass at CT, the presence of a tumoral vessel extending into the renal cortex or a renal parenchymal defect at the site of tumor contact strongly favors the diagnosis of exophytic angiomyolipoma, whereas calcifications suggest liposarcoma.


Immunotherapy of prostate cancer in the Dunning rat model: use of cytokine gene modified tumor vaccines.
Adenocarcinoma of the prostate is the most common cancer in men. The majority of cancers are discovered once they have already metastasized, and there is no effective therapy for prostatic cancer at this stage. The use of cytokine-secreting tumor cell preparations as therapeutic vaccines for the treatment of advanced prostate cancer was investigated in the Dunning rat R3327-MatLyLu prostatic tumor model. IL-2 secreting, irradiated, tumor cell preparations were capable of curing animals with s.c. established tumors, and induced immunological memory that protected animals from subsequent tumor challenge. Immunotherapy was less effective when tumors were induced orthotopically, but nevertheless led to improved outcome, significantly delaying, and occasionally preventing, recurrence of tumors after resection of the cancerous prostate. Granulocyte-macrophage colony stimulating factor secreting tumor cell preparations were less effective, and interferon-gamma secreting cells had only a marginal effect. Induction of a potent immune response in tumor bearing animals against the nonimmunogenic MatLyLu tumor supports the view that active immunotherapy warrants further investigation as a potential therapeutic approach to prostate cancer.


A dual-signal sensing system based on organic dyes-LDHs film for fluorescence detection of cysteine.
It is of great significance to sense cysteine (Cys) in a simple and fast way because of its potential applications in biological processes. In this contribution, we proposed a Cu2+-mediated fluorescence switchable strategy to detect Cys by designing a dual-signal fluorescence-enhanced sensor. Two organic dyes immobilized onto layered double hydroxides (LDHs) as novel sensors have been fabricated. It was found that the two fluorescence emissions of the obtained ultrathin films (donated as calcein@NFR/LDHs UTFs) could be effectively quenched by Cu2+ (off state), then reversibly recovered by Cys (on state) owing to the specific coordination of Cys and Cu2+. Under the optimal conditions, there were good linear relationships between the off-on efficiency (off-on) and the amount of Cys in the range of 1.0 × 10-7-1.2 × 10-6 M, with lower detection limits of 5.0 × 10-8 M at 520 nm and 2.3 × 10-8 M at 607 nm, respectively. There is little interference by common ions, amino acids and some excipients. The dual-signal fluorescence-enhanced response of the sensor contributes to its high sensitivity and selectivity. The proposed strategy is label-free, fast and selective, which is applicable to the analysis of Cys in medicine with satisfactory results.


Identification of Multiple Hard X-Ray Sources in Solar Flares: A Bayesian Analysis of the 2002 February 20 Event.
The hard X-ray emission in a solar flare is typically characterized by a number of discrete sources, each with its own spectral, temporal, and spatial variability. Establishing the relationship among these sources is critical to determining the role of each in the energy release and transport processes that occur within the flare. In this paper we present a novel method to identify and characterize each source of hard X-ray emission. The method permits a quantitative determination of the most likely number of subsources present, and of the relative probabilities that the hard X-ray emission in a given subregion of the flare is represented by a complicated multiple source structure or by a simpler single source. We apply the method to a well-studied flare on 2002 February 20 in order to assess competing claims as to the number of chromospheric footpoint sources present, and hence to the complexity of the underlying magnetic geometry/topology. Contrary to previous claims of the need for multiple sources to account for the chromospheric hard X-ray emission at different locations and times, we find that a simple two-footpoint-plus-coronal-source model is the most probable explanation for the data. We also find that one of the footpoint sources moves quite rapidly throughout the event, a factor that presumably complicated previous analyses. The inferred velocity of the footpoint corresponds to a very high induced electric field, compatible with the fields in thin reconnecting current sheets.


Trophoblast stem cells.
At the first cell fate decision in mammalian development, the origins of trophoblast and embryonic cell lineages are established as the trophectoderm and the inner cell mass (ICM) in the blastocyst. In the trophoblast cell lineage, a subset of the trophectoderm cells maintains the capacity to proliferate and contribute to the extraembryonic ectoderm, the ectoplacental cone, and the secondary giant cells of the early conceptus after implantation, and finally they produce the entire trophoblastic population in the mature placenta. The stem cell population of the trophectoderm lineage can be isolated and maintained in vitro in the presence of fibroblast growth factor 4, heparin, and a feeder layer of mouse embryonic fibroblast cells. These apparently immortal stem cells in culture are termed trophoblast stem (TS) cells, and exhibit the potential to differentiate into multiple trophoblastic cell types in vitro, as well as in vivo. Even after multiple passages, TS cells retain the ability to participate in the normal development of chimeras and contribute exclusively to the trophoblastic component of the placenta and of the parietal yolk sac. The fate of TS cells is strikingly in contrast to that of embryonic stem cells, which never contribute to these tissues. In this chapter, detailed protocols for the isolation and establishment of TS cell lines from blastocysts and their maintenance are described.


Genomic Selection and Genome-wide Association Study for Feed-Efficiency Traits in a Farmed Nile Tilapia (Oreochromis niloticus) Population.
Nile tilapia is a key aquaculture species with one of the highest production volumes globally. Genetic improvement of feed efficiency via selective breeding is an important goal, and genomic selection may expedite this process. The aims of this study were to 1) dissect the genetic architecture of feed-efficiency traits in a Nile tilapia breeding population, 2) map the genomic regions associated with these traits and identify candidate genes, 3) evaluate the accuracy of breeding value prediction using genomic data, and 4) assess the impact of the genetic marker density on genomic prediction accuracies. Using an experimental video recording trial, feed conversion ratio (FCR), body weight gain (BWG), residual feed intake (RFI) and feed intake (FI) traits were recorded in 40 full-sibling families from the GIFT (Genetically Improved Farmed Tilapia) Nile tilapia breeding population. Fish were genotyped with a ThermoFisher Axiom 65 K Nile tilapia SNP array. Significant heritabilities, ranging from 0.12 to 0.22, were estimated for all the assessed traits using the genomic relationship matrix. A negative but favourable genetic correlation was found between BWG and the feed-efficiency related traits; -0.60 and -0.63 for FCR and RFI, respectively. While the genome-wide association analyses suggested a polygenic genetic architecture for all the measured traits, there were significant QTL identified for BWG and FI on chromosomes seven and five respectively. Candidate genes previously found to be associated with feed-efficiency traits were located in these QTL regions, including ntrk3a, ghrh and eif4e3. The accuracy of breeding value prediction using the genomic data was up to 34% higher than using pedigree records. A SNP density of approximately 5,000 SNPs was sufficient to achieve similar prediction accuracy as the full genotype data set. Our results highlight the potential of genomic selection to improve feed efficiency traits in Nile tilapia breeding programmes.


Hyper IgM syndrome presenting as chronic suppurative lung disease.
The Hyper-immunoglobulin M syndromes (HIGM) are a heterogeneous group of genetic disorders resulting in defects of immunoglobulin class switch recombination. Affected patients show humoral immunodeficiency and high susceptibility to opportunistic infections. Elevated serum IgM levels are the hallmark of the disease, even though in few rare cases they may be in the normal range. Hyper IgM is associated with low to undetectable levels of serum IgG, IgA, and IgE. In some cases, alterations in different genes may be identified. Mutations in five genes have so far been associated to the disease, which can be inherited with an X-linked (CD40 ligand, and nuclear factor-kB essential modulator defects) or an autosomal recessive (CD40, activation-induced cytidine deaminase, and uracil-DNA glycosylase mutation) pattern. The patient herein described presented with recurrent upper and lower respiratory infections and evidence of suppurative lung disease at the conventional chest imaging. The presence of low serum IgG and IgA levels, elevated IgM levels, and a marked reduction of in vivo switched memory B cells led to a clinical and functional diagnosis of HIGM although the genetic cause was not identified.


ATPase active-site electrostatic interactions control the global conformation of the 100 kDa SecA translocase.
SecA is an intensively studied mechanoenzyme that uses ATP hydrolysis to drive processive extrusion of secreted proteins through a protein-conducting channel in the cytoplasmic membrane of eubacteria. The ATPase motor of SecA is strongly homologous to that in DEAD-box RNA helicases. It remains unclear how local chemical events in its ATPase active site control the overall conformation of an ~100 kDa multidomain enzyme and drive protein transport. In this paper, we use biophysical methods to establish that a single electrostatic charge in the ATPase active site controls the global conformation of SecA. The enzyme undergoes an ATP-modulated endothermic conformational transition (ECT) believed to involve similar structural mechanics to the protein transport reaction. We have characterized the effects of an isosteric glutamate-to-glutamine mutation in the catalytic base, a mutation which mimics the immediate electrostatic consequences of ATP hydrolysis in the active site. Calorimetric studies demonstrate that this mutation facilitates the ECT in Escherichia coli SecA and triggers it completely in Bacillus subtilis SecA. Consistent with the substantial increase in entropy observed in the course of the ECT, hydrogen-deuterium exchange mass spectrometry demonstrates that it increases protein backbone dynamics in domain-domain interfaces at remote locations from the ATPase active site. The catalytic glutamate is one of ~250 charged amino acids in SecA, and yet neutralization of its side chain charge is sufficient to trigger a global order-disorder transition in this 100 kDa enzyme. The intricate network of structural interactions mediating this effect couples local electrostatic changes during ATP hydrolysis to global conformational and dynamic changes in SecA. This network forms the foundation of the allosteric mechanochemistry that efficiently harnesses the chemical energy stored in ATP to drive complex mechanical processes.