Evidence for adipocyte-derived extracellular vesicles in the human circulation

Adipocyte-derived extracellular vesicles (EVs) may serve as novel endocrine mediators of adipose tissue and impact upon vascular health. However, it is unclear whether adipocyte-derived EVs are present in the human circulation. Therefore, the purpose of this study was to seek evidence for the presence of adipocyte-derived EVs in circulating plasma. Size exclusion chromatography of platelet-free plasma identified fractions 5-10 as containing EVs by a peak in particle concentration, which corresponded with the presence of EV and adipocyte proteins. Pooling fractions 5-10 and subjecting to ultracentrifugation yielded a plasma EV sample, as verified by transmission electron microscopy (TEM) showing EV structures and Western blotting for EV (e.g. CD9 and Alix) and adipocyte markers. Magnetic beads and a solid phase assay were used to deplete the EV sample of the four major families of circulating EVs: platelet-, leukocyte-, endothelial- and erythrocyte-derived EVs. Post-depletion samples from both techniques contained EV structures as visualized by TEM, as well as CD9, Alix and classic adipocyte proteins. Post-depletion samples also contained a range of other adipocyte proteins from an adipokine array. Adipocyte proteins and adipokines are expressed in optimally processed plasma EV samples, suggesting that adipocyte-derived EVs are secreted into the human circulation

Characterisation of adipocyte-derived extracellular vesicles released pre- and post-adipogenesis

Extracellular vesicles (EVs) are submicron vesicles released from many cell types, including adipocytes. EVs are implicated in the pathogenesis of obesity-driven cardiovascular disease, although the characteristics of adipocyte-derived EVs are not well described. We sought to define the characteristics of adipocyte-derived EVs before and after adipogenesis, hypothesising that adipogenesis would affect EV structure, molecular composition and function. Using 3T3-L1 cells, EVs were harvested at day 0 and day 15 of differentiation. EV and cell preparations were visualised by electron microscopy and EVs quantified by nanoparticle tracking analysis (NTA). EVs were then assessed for annexin V positivity using flow cytometry; lipid and phospholipid composition using 2D thin layer chromatography and gas chromatography; and vesicular protein content by an immuno-phenotyping assay. Pre-adipogenic cells are connected via a network of protrusions and EVs at both time points display classic EV morphology. EV concentration is elevated prior to adipogenesis, particularly in exosomes and small microvesicles. Parent cells contain higher proportions of phosphatidylserine (PS) and show higher annexin V binding. Both cells and EVs contain an increased proportion of arachidonic acid at day 0. PREF-1 was increased at day 0 whilst adiponectin was higher at day 15 indicating EV protein content reflects the stage of adipogenesis of the cell. Our data suggest that EV production is higher in cells before adipogenesis, particularly in vesicles <300 nm. Cells at this time point possess a greater proportion of PS (required for EV generation) whilst corresponding EVs are enriched in signalling fatty acids, such as arachidonic acid, and markers of adipogenesis, such as PREF-1 and PPARγ

Health screenings in college health centers: Variations in practice

Objective: The purpose of this study was to examine the routine screening of female students in college health centers for six priority health-related behaviors and experiences (tobacco use, alcohol use, eating disorders [EDs], obesity, anxiety and depression, intimate partner violence/sexual violence [IPV/SV]), and to identify variations in practice. Participants: A nationally representative sample of 1,221 healthcare providers (HCPs), including nurse practitioners, physicians, and physician assistants, from 471 U.S. college health centers. Methods: HCPs completed surveys (on-line or paper) and reported on routine screening of female college students. Results: HCPs reported consistently high rates (75–85%) of screening for tobacco use, alcohol use, and anxiety/depression. Rates of screening for IPV/SV, obesity and EDs were much lower. Nurse practitioners reported the highest IPV/SV screening rates. Conclusions: College health centers present unique opportunities for screening, case-finding and intervening to reduce long-term sequelae. Providers are well-positioned to lead initiatives to improve screening practices

The care of patients with Duchenne, Becker and other muscular dystrophies in the COVID-19 pandemic

The corona virus disease 2019 (COVID-19) pandemic has resulted in the reorganization of healthcare settings affecting clinical care delivery to patients with Duchenne and Becker muscular dystrophy (DBMD) as well as other inherited muscular dystrophies. The magnitude of the impact of this public health emergency on the care of patients with DBMD is unclear as they are suspected of having an increased risk for severe manifestations of COVID-19. In this paper, the authors discuss their consensus recommendations pertaining to care of these patients during the pandemic. We address issues surrounding corticosteroid and exon skipping treatments, cardiac medications, hydroxychloroquine use, emergency/respiratory care, rehabilitation management, and the conduct of clinical trials. We highlight the importance of collaborative treatment decisions between the patient, family, and health care provider, considering any geographic or institution-specific policies and precautions for COVID-19. We advocate for continuing multidisciplinary care for these patients using telehealth

The DEEP Groth Strip Survey. I. The Sample

The Deep Extragalactic Exploratory Probe (DEEP) is a multi-phase research program dedicated to the study of the formation and evolution of galaxies and of large scale structure in the distant Universe. This paper describes the first five-year phase, denoted DEEP1. A series of ten DEEP1 papers will discuss a range of scientific topics (e.g., the study of photometric and spectral properties of a general distant galaxy survey, the evolution observed in galaxy populations of varied morphologies). The observational basis for these studies is the Groth Survey Strip field, a 127 square arcminute region which has been observed with the Hubble Space Telescope in both broad I-band and V-band optical filters and with the Low Resolution Imaging Spectrograph on the Keck Telescopes. Catalogs of photometric and structural parameters have been constructed for 11,547 galaxies and stars at magnitudes brighter than 29, and spectroscopy has been conducted for a magnitude-color weighted subsample of 818 objects. We evaluate three independent techniques for constructing an imaging catalog for the field from the HST data, and discuss the depth and sampling of the resultant catalogs. The selection of the spectroscopic subsample is discussed, and we describe the multifaceted approach taken to prioritizing objects of interest for a variety of scientific subprograms. A series of Monte Carlo simulations then demonstrates that the spectroscopic subsample can be adequately modeled as a simple function of magnitude and color cuts in the imaging catalog.Comment: ApJS accepted, 15 pages, 12 figures. Version with higher-quality figures available at http://astronomy.nmsu.edu/nicol

Detection of group A Streptococcus in tonsils from pediatric patients reveals high rate of asymptomatic streptococcal carriage

<p>Abstract</p> <p>Background</p> <p>Group A <it>Streptococcus </it>(GAS) causes acute tonsillopharyngitis in children, and approximately 20% of this population are chronic carriers of GAS. Antibacterial therapy has previously been shown to be insufficient at clearing GAS carriage. Bacterial biofilms are a surface-attached bacterial community that is encased in a matrix of extracellular polymeric substances. Biofilms have been shown to provide a protective niche against the immune response and antibiotic treatments, and are often associated with recurrent or chronic bacterial infections. The objective of this study was to test the hypothesis that GAS is present within tonsil tissue at the time of tonsillectomy.</p> <p>Methods</p> <p>Blinded immunofluorescent and histological methods were employed to evaluate palatine tonsils from children undergoing routine tonsillectomy for adenotonsillar hypertrophy or recurrent GAS tonsillopharyngitis.</p> <p>Results</p> <p>Immunofluorescence analysis using anti-GAS antibody was positive in 11/30 (37%) children who had tonsillectomy for adenotonsillar hypertrophy and in 10/30 (33%) children who had tonsillectomy for recurrent GAS pharyngitis. Fluorescent microscopy with anti-GAS and anti-cytokeratin 8 & 18 antibodies revealed GAS was localized to the tonsillar reticulated crypts. Scanning electron microscopy identified 3-dimensional communities of cocci similar in size and morphology to GAS. The characteristics of these communities are similar to GAS biofilms from <it>in vivo </it>animal models.</p> <p>Conclusion</p> <p>Our study revealed the presence of GAS within the tonsillar reticulated crypts of approximately one-third of children who underwent tonsillectomy for either adenotonsillar hypertrophy or recurrent GAS tonsillopharyngitis at the Wake Forest School of Medicine.</p> <p>Trial Registration</p> <p>The tissue collected was normally discarded tissue and no patient identifiers were collected. Thus, no subjects were formally enrolled.</p

Molecular tools for bathing water assessment in Europe:Balancing social science research with a rapidly developing environmental science evidence-base

The use of molecular tools, principally qPCR, versus traditional culture-based methods for quantifying microbial parameters (e.g., Fecal Indicator Organisms) in bathing waters generates considerable ongoing debate at the science-policy interface. Advances in science have allowed the development and application of molecular biological methods for rapid (~2 h) quantification of microbial pollution in bathing and recreational waters. In contrast, culture-based methods can take between 18 and 96 h for sample processing. Thus, molecular tools offer an opportunity to provide a more meaningful statement of microbial risk to water-users by providing near-real-time information enabling potentially more informed decision-making with regard to water-based activities. However, complementary studies concerning the potential costs and benefits of adopting rapid methods as a regulatory tool are in short supply. We report on findings from an international Working Group that examined the breadth of social impacts, challenges, and research opportunities associated with the application of molecular tools to bathing water regulations

Lipoprotein-apheresis reduces circulating microparticles in individuals with familial hypercholesterolemia

Lipoprotein-apheresis (apheresis) removes LDL-cholesterol in patients with severe dyslipidemia. However, reduction is transient, indicating that the long-term cardiovascular benefits of apheresis may not solely be due to LDL removal. Microparticles (MPs) are submicron vesicles released from the plasma membrane of cells. MPs, particularly platelet-derived MPs, are increasingly being linked to the pathogenesis of many diseases. We aimed to characterize the effect of apheresis on MP size, concentration, cellular origin, and fatty acid concentration in individuals with familial hypercholesterolemia (FH). Plasma and MP samples were collected from 12 individuals with FH undergoing routine apheresis. Tunable resistive pulse sensing (np200) and nanoparticle tracking analysis measured a fall in MP concentration (33 and 15%, respectively; P < 0.05) pre- to post-apheresis. Flow cytometry showed MPs were predominantly annexin V positive and of platelet (CD41) origin both pre- (88.9%) and post-apheresis (88.4%). Fatty acid composition of MPs differed from that of plasma, though apheresis affected a similar profile of fatty acids in both compartments, as measured by GC-flame ionization detection. MP concentration was also shown to positively correlate with thrombin generation potential. In conclusion, we show apheresis nonselectively removes annexin V-positive platelet-derived MPs in individuals with FH. These MPs are potent inducers of coagulation and are elevated in CVD; this reduction in pathological MPs could relate to the long-term benefits of apheresis

Expression of Activated PIK3CA in Ovarian Surface Epithelium Results in Hyperplasia but Not Tumor Formation

activation is one of the early genetic events in ovarian cancer. However, its role in malignant transformation of ovarian surface epithelium (OSE) is largely unclear..

An exploration of the knowledge and attitudes towards breastfeeding among a sample of Chinese mothers in Ireland

<p>Abstract</p> <p>Background</p> <p>Psychological factors are important in influencing breastfeeding practices. This retrospective study explored knowledge and attitudes related to breastfeeding of Chinese mothers living in Ireland.</p> <p>Methods</p> <p>A cross-sectional self-administrated survey written in Chinese was distributed to a convenience sample of 322 immigrant Chinese mothers mainly <it>via </it>Chinese supermarkets and Chinese language schools in Dublin, with the involvement of the snowball method to increase sample size. Maternal breastfeeding knowledge and attitudes were described, their associations with socio-demographic variables were explored by Chi-square analysis, and their independent associations with breastfeeding behaviours were estimated by binary logistic regression analyses.</p> <p>Results</p> <p>In spite of considerable awareness of the advantages of breastfeeding (mean score = 4.03 ± 0.73), some misconceptions (<it>e</it>.<it>g</it>. 'mother should stop breastfeeding if she catches a cold') and negative attitudes (<it>e</it>.<it>g</it>. breastfeeding inconvenient, embarrassing, and adverse to mothers' figure) existed, especially among the less educated mothers. Cultural beliefs concerning the traditional Chinese postpartum diet were prevalent, particularly among those who had lived in Ireland for a shorter duration (P = 0.004). Psychological parameters had strong independent associations with breastfeeding practices in this study. Those who had lower awareness score (OR = 2.98, 95% CI: 1.87-4.73), more misconceptions and negative attitudes (P < 0.05), and weaker cultural beliefs (P < 0.05) were less likely to breastfeed.</p> <p>Conclusions</p> <p>Findings highlight a need to focus resources and education on correcting the misconceptions identified and reversing the negative attitudes towards breastfeeding among Chinese mothers in Ireland, in particular those with primary/secondary level of education. Mothers' cultural beliefs should also be acknowledged and understood by healthcare providers.</p