Zoom in on antibody aggregates:A potential pitfall in the search of rare EV populations

High-resolution flow cytometers (hFCM) are used for the detection of extracellular vesicles (EV) in various biological fluids. Due to the increased sensitivity of hFCM, new artifacts with the potential of interfering with data interpretation are introduced, such as detection of antibody aggregates. The aim of this study was to investigate the extent of aggregates in labels commonly used for the characterization of EVs by hFCM. Furthermore, we aimed to compare the efficacy of centrifugation and filtering treatments to remove aggregates, as well as to quantify the effect of the treatments in reducing aggregates. For this purpose, we labeled phosphate buffered saline (PBS) with fluorescently conjugated protein labels and antibodies after submitting them to 5, 10, or 30 min centrifugation, filtering or washed filtering. We investigated samples by hFCM and quantified the amount of aggregates found in PBS labeled with untreated and pre-treated labels. We found a varying amount of aggregates in all labels investigated, and further that filtering is most efficient in removing all but the smallest aggregates. Filtering protein labels can reduce the extent of aggregates; however, how much remains depends on the specific labels and their combination. Therefore, it is still necessary to include appropriate controls in a hFCM study of EVs

Oscillation mode linewidths and heights of 23 main-sequence stars observed by Kepler

Solar-like oscillations have been observed by Kepler and CoRoT in many solar-type stars, thereby providing a way to probe the stars using asteroseismology. We provide the mode linewidths and mode heights of the oscillations of various stars as a function of frequency and of effective temperature. We used a time series of nearly two years of data for each star. The 23 stars observed belong to the simple or F-like category. The power spectra of the 23 main-sequence stars were analysed using both maximum likelihood estimators and Bayesian estimators, providing individual mode characteristics such as frequencies, linewidths, and mode heights. We study the source of systematic errors in the mode linewidths and mode heights, and we present a way to correct these errors with respect to a common reference fit. Using the correction, we could explain all sources of systematic errors, which could be reduced to less than $\pm$15% for mode linewidths and heights, and less than $\pm$5% for amplitude, when compared to the reference fit. The effect of a different estimated stellar background and a different estimated splitting will provide frequency-dependent systematic errors that might affect the comparison with theoretical mode linewidth and mode height, therefore affecting the understanding of the physical nature of these parameters. All other sources of relative systematic errors are less dependent upon frequency. We also provide the dependence of the so-called linewidth dip, in the middle of the observed frequency range, as a function of effective temperature. We show that the depth of the dip decreases with increasing effective temperature. The dependence of the dip on effective temperature may imply that the mixing length parameter $\alpha$ or the convective flux may increase with effective temperature.Comment: Accepted by A&A, 38 pages, 35 figures, 26 table

First introduction of highly pathogenic H5N1 avian influenza A viruses in wild and domestic birds in Denmark, Northern Europe

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licens

A randomized, placebo-controlled trial of late Na current inhibition (ranolazine) in coronary microvascular dysfunction (CMD): impact on angina and myocardial perfusion reserve.

AimsThe mechanistic basis of the symptoms and signs of myocardial ischaemia in patients without obstructive coronary artery disease (CAD) and evidence of coronary microvascular dysfunction (CMD) is unclear. The aim of this study was to mechanistically test short-term late sodium current inhibition (ranolazine) in such subjects on angina, myocardial perfusion reserve index, and diastolic filling.Materials and resultsRandomized, double-blind, placebo-controlled, crossover, mechanistic trial in subjects with evidence of CMD [invasive coronary reactivity testing or non-invasive cardiac magnetic resonance imaging myocardial perfusion reserve index (MPRI)]. Short-term oral ranolazine 500-1000 mg twice daily for 2 weeks vs. placebo. Angina measured by Seattle Angina Questionnaire (SAQ) and SAQ-7 (co-primaries), diary angina (secondary), stress MPRI, diastolic filling, quality of life (QoL). Of 128 (96% women) subjects, no treatment differences in the outcomes were observed. Peak heart rate was lower during pharmacological stress during ranolazine (-3.55 b.p.m., P < 0.001). The change in SAQ-7 directly correlated with the change in MPRI (correlation 0.25, P = 0.005). The change in MPRI predicted the change in SAQ QoL, adjusted for body mass index (BMI), prior myocardial infarction, and site (P = 0.0032). Low coronary flow reserve (CFR <2.5) subjects improved MPRI (P < 0.0137), SAQ angina frequency (P = 0.027), and SAQ-7 (P = 0.041).ConclusionsIn this mechanistic trial among symptomatic subjects, no obstructive CAD, short-term late sodium current inhibition was not generally effective for SAQ angina. Angina and myocardial perfusion reserve changes were related, supporting the notion that strategies to improve ischaemia should be tested in these subjects.Trial registrationclinicaltrials.gov Identifier: NCT01342029

An Introduction to Data Analysis in Asteroseismology

A practical guide is presented to some of the main data analysis concepts and techniques employed contemporarily in the asteroseismic study of stars exhibiting solar-like oscillations. The subjects of digital signal processing and spectral analysis are introduced first. These concern the acquisition of continuous physical signals to be subsequently digitally analyzed. A number of specific concepts and techniques relevant to asteroseismology are then presented as we follow the typical workflow of the data analysis process, namely, the extraction of global asteroseismic parameters and individual mode parameters (also known as peak-bagging) from the oscillation spectrum.Comment: Lecture presented at the IVth Azores International Advanced School in Space Sciences on "Asteroseismology and Exoplanets: Listening to the Stars and Searching for New Worlds" (arXiv:1709.00645), which took place in Horta, Azores Islands, Portugal in July 201

Limits on surface gravities of Kepler planet-candidate host stars from non-detection of solar-like oscillations

We present a novel method for estimating lower-limit surface gravities log g of Kepler targets whose data do not allow the detection of solar-like oscillations. The method is tested using an ensemble of solar-type stars observed in the context of the Kepler Asteroseismic Science Consortium. We then proceed to estimate lower-limit log g for a cohort of Kepler solar-type planet-candidate host stars with no detected oscillations. Limits on fundamental stellar properties, as provided by this work, are likely to be useful in the characterization of the corresponding candidate planetary systems. Furthermore, an important byproduct of the current work is the confirmation that amplitudes of solar-like oscillations are suppressed in stars with increased levels of surface magnetic activity.Comment: Accepted for publication in ApJ; 35 pages, 10 figures, 5 table

Oscillation mode frequencies of 61 main sequence and subgiant stars observed by Kepler

Solar-like oscillations have been observed by Kepler and CoRoT in several solar-type stars, thereby providing a way to probe the stars using asteroseismology. We provide the mode frequencies of the oscillations of various stars required to perform a comparison with those obtained from stellar modelling. We used a time series of nine months of data for each star. The 61 stars observed were categorised in three groups: simple, F-like and mixed-mode. The simple group includes stars for which the identification of the mode degree is obvious. The F-like group includes stars for which the identification of the degree is ambiguous. The mixed-mode group includes evolved stars for which the modes do not follow the asymptotic relation of low-degree frequencies. Following this categorisation, the power spectra of the 61 main sequence and subgiant stars were analysed using both maximum likelihood estimators and Bayesian estimators, providing individual mode characteristics such as frequencies, linewidths, and mode heights. We developed and describe a methodology for extracting a single set of mode frequencies from multiple sets derived by different methods and individual scientists. We report on how one can assess the quality of the fitted parameters using the likelihood ratio test and the posterior probabilities. We provide the mode frequencies of 61 stars (with their 1-sigma error bars), as well as their associated echelle diagrams.Comment: 83 pages, 17 figures, 61 tables, paper accepted by Astronomy and Astrophysic

Circulating exosomes deliver free fatty acids from the bloodstream to cardiac cells:Possible role of CD36

Regulation of circulating free fatty acid (FFA) levels and delivery is crucial to maintain tissue homeostasis. Exosomes are nanomembranous vesicles that are released from diverse cell types and mediate intercellular communication by delivering bioactive molecules. Here, we sought to investigate the uptake of FFAs by circulating exosomes, the delivery of FFA-loaded exosomes to cardiac cells and the possible role of the FFA transporter CD36 in these processes. Circulating exosomes were purified from the serum of healthy donors after an overnight fast (F) or 20 minutes after a high caloric breakfast (postprandial, PP). Western blotting, Immunogold Electron Microscopy and FACS analysis of circulating exosomes showed that CD36 was expressed under both states, but was higher in postprandial-derived exosomes. Flow cytometry analysis showed that circulating exosomes were able to take-up FFA directly from serum. Importantly, preincubation of exosomes with a blocking CD36 antibody significantly impeded uptake of the FFA analogue BODIPY, pointing to the role of CD36 in FFA exosomal uptake. Finally, we found that circulating exosomes could delivery FFA analogue BODIPY into cardiac cells ex vivo and in vivo in a mice model. Overall, our results suggest a novel mechanism in which circulating exosomes can delivery FFAs from the bloodstream to cardiac tissue. Further studies will be necessary to understand this mechanism and, in particular, its potential involvement in metabolic pathologies such as obesity, diabetes and atherosclerosis