Cohesin controls X chromosome structure remodeling and X-reactivation during mouse iPSC-reprogramming.

Reactivation of the inactive X chromosome is a hallmark epigenetic event during reprogramming of mouse female somatic cells to induced pluripotent stem cells (iPSCs). This involves global structural remodeling from a condensed, heterochromatic into an open, euchromatic state, thereby changing a transcriptionally inactive into an active chromosome. Despite recent advances, very little is currently known about the molecular players mediating this process and how this relates to iPSC-reprogramming in general. To gain more insight, here we perform a RNAi-based knockdown screen during iPSC-reprogramming of mouse fibroblasts. We discover factors important for X chromosome reactivation (XCR) and iPSC-reprogramming. Among those, we identify the cohesin complex member SMC1a as a key molecule with a specific function in XCR, as its knockdown greatly affects XCR without interfering with iPSC-reprogramming. Using super-resolution microscopy, we find SMC1a to be preferentially enriched on the active compared with the inactive X chromosome and that SMC1a is critical for the decompacted state of the active X. Specifically, depletion of SMC1a leads to contraction of the active X both in differentiated and in pluripotent cells, where it normally is in its most open state. In summary, we reveal cohesin as a key factor for remodeling of the X chromosome from an inactive to an active structure and that this is a critical step for XCR during iPSC-reprogramming.

Relational diversity in social portfolios predicts well-being.

We document a link between the relational diversity of one's social portfolio-the richness and evenness of relationship types across one's social interactions-and well-being. Across four distinct samples, respondents from the United States who completed a preregistered survey (n = 578), respondents to the American Time Use Survey (n = 19,197), respondents to the World Health Organization's Study on Global Aging and Adult Health (n = 10,447), and users of a French mobile application (n = 21,644), specification curve analyses show that the positive relationship between social portfolio diversity and well-being is robust across different metrics of well-being, different categorizations of relationship types, and the inclusion of a wide range of covariates. Over and above people's total amount of social interaction and the diversity of activities they engage in, the relational diversity of their social portfolio is a unique predictor of well-being, both between individuals and within individuals over time.

Inequality in socially permissible consumption.

Lower-income individuals are frequently criticized for their consumption decisions; this research examines why. Eleven preregistered studies document systematic differences in permissible consumption-interpersonal judgments about what is acceptable (or not) for others to consume-such that lower-income individuals' decisions are subject to more negative and restrictive evaluations. Indeed, the same consumption decisions may be deemed less permissible for a lower-income individual than for an individual with higher or unknown income (studies 1A and 1B), even when purchased with windfall funds. This gap persists among participants from a large, nationally representative sample (study 2) and when testing a broad array of "everyday" consumption items (study 3). Additional studies investigate why: The same items are often perceived as less necessary for lower- (versus higher-) income individuals (studies 4 and 5). Combining both permissibility and perceived necessity, additional studies (studies 6 and 7) demonstrate a causal link between the two constructs: A purchase decision will be deemed permissible (or not) to the extent that it is perceived as necessary (or not). However, because-for lower-income individuals-fewer items are perceived as necessary, fewer are therefore socially permissible to consume. This finding not only exposes a fraught double standard, but also portends consequential behavioral implications: People prefer to allocate strictly "necessary" items to lower-income recipients (study 8), even if such items are objectively and subjectively less valuable (studies 9A and 9B), which may result in an imbalanced and inefficient provision of resources to the poor.

A conserved expression signature predicts growth rate and reveals cell & lineage-specific differences.

Isogenic cells cultured together show heterogeneity in their proliferation rate. To determine the differences between fast and slow-proliferating cells, we developed a method to sort cells by proliferation rate, and performed RNA-seq on slow and fast proliferating subpopulations of pluripotent mouse embryonic stem cells (mESCs) and mouse fibroblasts. We found that slowly proliferating mESCs have a more naïve pluripotent character. We identified an evolutionarily conserved proliferation-correlated transcriptomic signature that is common to all eukaryotes: fast cells have higher expression of genes for protein synthesis and protein degradation. This signature accurately predicted growth rate in yeast and cancer cells, and identified lineage-specific proliferation dynamics during development, using C. elegans scRNA-seq data. In contrast, sorting by mitochondria membrane potential revealed a highly cell-type specific mitochondria-state related transcriptome. mESCs with hyperpolarized mitochondria are fast proliferating, while the opposite is true for fibroblasts. The mitochondrial electron transport chain inhibitor antimycin affected slow and fast subpopulations differently. While a major transcriptional-signature associated with cell-to-cell heterogeneity in proliferation is conserved, the metabolic and energetic dependency of cell proliferation is cell-type specific.

Frequency and Mechanisms of Spontaneous Fosfomycin Nonsusceptibility Observed upon Disk Diffusion Testing of Escherichia coli.

Fosfomycin maintains activity against most Escherichia coli clinical isolates, but the growth of E. coli colonies within the zone of inhibition around the fosfomycin disk is occasionally observed upon susceptibility testing. We aimed to estimate the frequency of such nonsusceptible inner colony mutants and identify the underlying resistance mechanisms. Disk diffusion testing of fosfomycin was performed on 649 multidrug-resistant E. coli clinical isolates collected between 2011 and 2015. For those producing inner colonies inside the susceptible range, the parental strains and their representative inner colony mutants were subjected to MIC testing, whole-genome sequencing, reverse transcription-quantitative PCR (qRT-PCR), and carbohydrate utilization studies. Of the 649 E. coli clinical isolates, 5 (0.8%) consistently produced nonsusceptible inner colonies. Whole-genome sequencing revealed the deletion of uhpT encoding hexose-6-phosphate antiporter in 4 of the E. coli inner colony mutants, while the remaining mutant contained a nonsense mutation in uhpA The expression of uhpT was absent in the mutant strains with uhpT deletion and was not inducible in the strain with the uhpA mutation, unlike in its parental strain. All 5 inner colony mutants had reduced growth on minimal medium supplemented with glucose-6-phosphate. In conclusion, fosfomycin-nonsusceptible inner colony mutants can occur due to the loss of function or induction of UhpT but are rare among multidrug-resistant E. coli clinical strains. Considering that these mutants carry high biological costs, we suggest that fosfomycin susceptibility of strains that generate inner colony mutants can be interpreted on the basis of the zone of inhibition without accounting for the inner colonies.

Pharmacological folding chaperones act as allosteric ligands of Frizzled4.

Upon binding, ligands can chaperone their protein targets by preventing them from misfolding and aggregating. Thus, an organic molecule that works as folding chaperone for a protein might be its specific ligand, and, similarly, the chaperone potential could represent an alternative readout in a molecular screening campaign toward the identification of new hits. Here we show that small molecules selected for acting as pharmacological chaperones on a misfolded mutant of the Frizzled4 (Fz4) receptor bind and modulate wild-type Fz4, representing what are to our knowledge the first organic ligands of this until-now-undruggable GPCR. The novelty and the advantages of the screening platform, the allosteric binding site addressed by these new ligands and the mechanism they use to modulate Fz4 suggest new avenues for development of inhibitors of the Wnt-ß-catenin pathway and for drug discovery.

Comparison of Minocycline Susceptibility Testing Methods for Carbapenem-Resistant Acinetobacter baumannii.

Treatment options for infections due to carbapenem-resistant Acinetobacter baumannii are extremely limited. Minocycline is a semisynthetic tetracycline derivative with activity against this pathogen. This study compared susceptibility testing methods that are used in clinical microbiology laboratories (Etest, disk diffusion, and Sensititre broth microdilution methods) for testing of minocycline, tigecycline, and doxycycline against 107 carbapenem-resistant A. baumannii clinical isolates. Susceptibility rates determined with the standard broth microdilution method using cation-adjusted Mueller-Hinton (MH) broth were 77.6% for minocycline and 29% for doxycycline, and 92.5% of isolates had tigecycline MICs of ≤2 µg/ml. Using MH agar from BD and Oxoid, susceptibility rates determined with the Etest method were 67.3% and 52.3% for minocycline, 21.5% and 18.7% for doxycycline, and 71% and 29.9% for tigecycline, respectively. With the disk diffusion method using MH agar from BD and Oxoid, susceptibility rates were 82.2% and 72.9% for minocycline and 34.6% and 34.6% for doxycycline, respectively, and rates of MICs of ≤2 µg/ml were 46.7% and 23.4% for tigecycline. In comparison with the standard broth microdilution results, very major rates were low (∼2.8%) for all three drugs across the methods, but major error rates were higher (∼5.6%), especially with the Etest method. For minocycline, minor error rates ranged from 14% to 37.4%. For tigecycline, minor error rates ranged from 6.5% to 69.2%. The majority of minor errors were due to susceptible results being reported as intermediate. For minocycline susceptibility testing of carbapenem-resistant A. baumannii strains, very major errors are rare, but major and minor errors overcalling strains as intermediate or resistant occur frequently with susceptibility testing methods that are feasible in clinical laboratories.

The acute management of ST-segment-elevation myocardial infarction.

Acute ST-segment-elevation myocardial infarction (STEMI) results from complete obstruction to coronary artery blood flow accompanied by the appearance of ST-segment-elevation on the electrocardiogram. Emergency treatment is required to restore coronary perfusion, thereby limiting the extent of damage to the myocardium and the likelihood of early death or future heart failure. This concise guideline summarises key recommendations from the National Institute for Health and Care Excellence clinical guideline on acute management of STEMI (CG167), of relevance to all healthcare professionals involved. Guidance is presented on choice of reperfusion strategies, procedural aspects, use of additional drugs before and alongside reperfusion therapies, and treatment of patients who are unconscious or in cardiogenic shock.

Control of dangerous substances in discharges and microbiological abatement: European framework and a case study of an ozone disinfection system.

Directive 2000/60/EC requires the achievement of a 'good chemical status' for surface water within pre-established dates. Disinfection is needed to achieve compulsory final microbial limit values (in Italy for wastewater discharges the parameter Escherichia coli - EC - is imposed by law with a maximum limit value of 5,000 cfu/100 mL). Liquid waste and disinfection by-products must be considered when designing appropriate monitoring of dangerous substances; the specific classes of substances must be investigated according to the typology of received wastewaters and liquid wastes (where applicable) and specific analytical techniques, with Limit of Detection (LOD) lower than the limit values, must be applied; the difficulties faced by national and regional environmental control Agencies is that these techniques have to be applied during ordinary activity and not only for research purposes. The study aims to present the control of dangerous substances, as a screening view, in wastewater treatment plant (WWTP) discharges in the province of Venice (Northern Italy) for the period 2007-2010 based on available data from institutional controls. In addition, the wastewater disinfection process with ozone applied to a medium size WWTP (45,000 Population Equivalents) is presented as a case study, with a view to assessing the microbiological abatement efficacy and the presence of dangerous substances. Discharge quality of the WWTPs in the province of Venice presented mean values that were higher than the LOD, but only for certain metals. For the Paese plant, zinc and chloroform were the only micro-pollutants detected with a higher level than the LOD. From microbiological data in the period 2006-2011 the disinfection abatement efficiency for Paese was, in most cases above 99% for EC, faecal coliform (FC), faecal streptococci (FS) while efficiency was slightly lower for total coliform (TC); however, the proposed criterion aimed at respecting 99.99% abatement was not completely satisfied. Therefore, despite the high organic and industrial load of the considered plant and the need to find an alternative system for chlorine, as chlorine disinfection has been banned in the Veneto region since December 2012, ozone efficiency is not completely satisfactory and other systems such as peracetic or performic acids and UV systems must be considered.

Corynebacterium kroppenstedtii breast abscesses in context, a retrospective cohort study.

Corynebacterium kroppenstedtii breast abscesses and granulomatous mastitis have gained increased recognition in the 20 years since their association was first described. No studies to date have described this organism in the context of all breast abscess pathogens. We retrospectively reviewed 160 community-acquired breast abscess samples from 135 patients in a 3 year period, describing the organisms isolated along with risk factors, site of infection and outcomes. We compared patient subgroups with acute and chronic abscesses, the latter defined as having a requirement for repeat aspiration more than 1 month apart. The prevalence of C. kroppenstedtii breast abscesses was 8 % in all patients (11/135), rising to 32 % in chronic abscesses (10/31), but only 1 % in acute abscesses (1/104; P<0.01). Only 10 % (1/10) of patients with C. kroppenstedtii chronic abscesses were smokers, whereas 75 % of patients (15/20) with non-C. kroppenstedtii chronic abscesses were smokers (P=0.01). C. kroppenstedtii should be considered in recurrent and prolonged infections, especially in non-smokers, and diagnostic methods altered accordingly. Identifying C. kroppenstedtii provides diagnostic clarity and alters management with recommendations for longer courses of treatment using non-beta-lactam antibiotics.

Video-Based Tracheostomy Care Education for Medical Students.

Objective: Tracheostomy is a common procedure that requires management by a multidisciplinary team of health care providers across a range of surgical and nonsurgical specialties. Nonsurgical health care providers have demonstrated a lack of knowledge and confidence in tracheostomy care, which improve with tracheostomy education programs. However, tracheostomy care is rarely included in preresidency medical education. The purpose of this study is to evaluate the effectiveness of a tracheostomy care video on third-year medical students' knowledge of and confidence in performing tracheostomy care. Methods: Prior to beginning clinical rotations, third-year medical students completed a 10-question tracheostomy care knowledge test (100 points total) and 11-question confidence survey (110 points total). After watching an 18-minute teaching video on tracheostomy care, students repeated the knowledge test and confidence survey. Results: An overall 147 medical students completed the educational module. After they watched the tracheostomy education video, their average score on the knowledge test improved from 57.8 to 88.9 out of 100 (P < .0001), and their average rating in confidence improved from 12.7 to 49.1 out of 110 (P < .0001). Students rated the helpfulness of the video a 7.4 out of 10. Discussion: Medical students' knowledge of tracheostomy care and confidence in caring for patients with tracheostomies improved after watching the video. Tracheostomy education should be included in early medical education so that future physicians of various specialties can better care for this patient population. Implications for Practice: Internet-published videos are an accessible educational resource with great potential application to various topics within otolaryngology, including tracheostomy care.

Revisiting the Landmark Task as a tool for studying hemispheric specialization: What's really right?

The "Landmark Task" (LT) is a line bisection judgment task that predominantly activates right parietal cortex. The typical version requires observers to judge bisections for horizontal lines that cross their egocentric midline and therefore may depend on spatial attention as well as spatial representation of the line segments. To ask whether the LT is indeed right-lateralized regardless of spatial attention (for which the right hemisphere is known to be important), we examined LT activation in 26 neurologically healthy young adults using vertical (instead of horizontal) stimuli, as compared with a luminance control task that made similar demands on spatial attention. We also varied task difficulty, which is known to affect lateralization in both spatial and language tasks. Despite these changes to the task, we observed right-lateralized parietal activations similar to those reported in other LT studies, both at group level and in individual lateralization indices. We conclude that LT activation is robustly right-lateralized, perhaps uniquely so among commonly-studied spatial tasks. We speculate that the unique properties of the LT reside in its requirement to judge relative magnitudes of the two line segments, rather than in the more general aspects of spatial attention or visual-spatial representation.

Current margin practice and effect on re-excision rates following the publication of the SSO-ASTRO consensus and ABS consensus guidelines: a national prospective study of 2858 women undergoing breast-conserving therapy in the UK and Ireland.

INTRODUCTION: There is variation in margin policy for breast conserving therapy (BCT) in the UK and Ireland. In response to the Society of Surgical Oncology and American Society for Radiation Oncology (SSO-ASTRO) margin consensus ('no ink on tumour' for invasive and 2 mm for ductal carcinoma in situ [DCIS]) and the Association of Breast Surgery (ABS) consensus (1 mm for invasive and DCIS), we report on current margin practice and unit infrastructure in the UK and Ireland and describe how these factors impact on re-excision rates. METHODS: A trainee collaborative-led multicentre prospective study was conducted in the UK and Ireland between 1st February and 31st May 2016. Data were collected on consecutive BCT patients and on local infrastructure and policies. RESULTS: A total of 79 sites participated in the data collection (75% screening units; average 372 cancers annually, range 70-900). For DCIS, 53.2% of units accept 1 mm and 38% accept 2-mm margins. For invasive disease 77.2% accept 1 mm and 13.9% accept 'no ink on tumour'. A total of 2858 patients underwent BCT with a mean re-excision rate of 17.2% across units (range 0-41%). The re-excision rate would be reduced to 15% if all units applied SSO-ASTRO guidelines and to 14.8% if all units followed ABS guidelines. Of those who required re-operation, 65% had disease present at margin. CONCLUSION: There continues to be large variation in margin policy and re-excision rates across units. Altering margin policies to follow either SSO-ASTRO or ABS guidelines would result in a modest reduction in the national re-excision rate. Most re-excisions are for involved margins rather than close margins.

Involvement of reactive oxygen species in the bactericidal activity of activated carbon fibre supporting silver; Bactericidal activity of ACF(Ag) mediated by ROS.

An activated carbon fibre supporting silver (ACF(Ag)) was tested for its antibacterial capacity against Escherichia coli (E. coli). Water that has passed through ACF(Ag) demonstrated strong bactericidal ability. This activity decreased over the time suggesting that generated bactericidal species were short lifespan. Since formation of reactive oxygen species (ROS) might be catalysed by silver impregnated and/or ACF itself, implication of ROS and silver was evaluated by the use of ROS scavengers and a silver ions neutralizing agent. The role of ROS in the E. coli mortality was confirmed by the use of a molecular approach which revealed a strong expression of oxidative stress genes.

Genome wide DNA copy number analysis of serous type ovarian carcinomas identifies genetic markers predictive of clinical outcome.

Ovarian cancer is the fifth leading cause of cancer death in women. Ovarian cancers display a high degree of complex genetic alterations involving many oncogenes and tumor suppressor genes. Analysis of the association between genetic alterations and clinical endpoints such as survival will lead to improved patient management via genetic stratification of patients into clinically relevant subgroups. In this study, we aim to define subgroups of high-grade serous ovarian carcinomas that differ with respect to prognosis and overall survival. Genome-wide DNA copy number alterations (CNAs) were measured in 72 clinically annotated, high-grade serous tumors using high-resolution oligonucleotide arrays. Two clinically annotated, independent cohorts were used for validation. Unsupervised hierarchical clustering of copy number data derived from the 72 patient cohort resulted in two clusters with significant difference in progression free survival (PFS) and a marginal difference in overall survival (OS). GISTIC analysis of the two clusters identified altered regions unique to each cluster. Supervised clustering of two independent large cohorts of high-grade serous tumors using the classification scheme derived from the two initial clusters validated our results and identified 8 genomic regions that are distinctly different among the subgroups. These 8 regions map to 8p21.3, 8p23.2, 12p12.1, 17p11.2, 17p12, 19q12, 20q11.21 and 20q13.12; and harbor potential oncogenes and tumor suppressor genes that are likely to be involved in the pathogenesis of ovarian carcinoma. We have identified a set of genetic alterations that could be used for stratification of high-grade serous tumors into clinically relevant treatment subgroups.

Systematic review of discriminating power of outcome measures used in clinical trials of fibromyalgia.

OBJECTIVE: Fibromyalgia (FM) comprises many symptoms and features. Consequently, studies on the condition have used a wide variety of outcome measures and assessment instruments. We investigated those outcome measures and instruments in association with the OMERACT (Outcome measures in Rheumatoid Arthritis Clinical Trials) FM Workshop initiative to define core outcome measures that should be used to assess FM. METHODS: A systematic literature review up to December 2007 was carried out using the keywords "fibromyalgia," "treatment" or "management," and "trial." Data were extracted on outcome measures and assessment instruments used and the pre and post mean and standard deviation to calculate effect sizes (ES). Further sensitivity analysis was carried out according to treatment type, blinding status, and study outcome. RESULTS: The outcome domains identified fell largely within those defined by OMERACT. Morning stiffness was frequently assessed and therefore has been included here. The number of assessment instruments used was wide-ranging, so sensitivity analysis was only carried out on the top 5 within each domain. ES ranged from 0.54 to 3.77 for the key OMERACT domains. Health-related quality of life (HRQOL) was the only exception that had no instrument with moderate sensitivity. Of the secondary domains, dyscognition was lacking any sensitive instrument, as were fatigue and anxiety in pharmacological trials. CONCLUSION: Each of the key OMERACT domains has an instrument that appears to be sensitive to change, with the exception of HRQOL, which requires further research. Dyscognition, fatigue, and anxiety would all benefit from more research into their assessment instruments.