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

M/L and Color Evolution for A Deep Sample of M* Cluster Galaxies at z~1: The Formation Epoch and the Tilt of the Fundamental Plane

We have measured velocity dispersions for a sample of 36 galaxies with J < 21.2 or Mr < -20.6 mag in MS1054-03, a massive cluster of galaxies at z = 0.83. Our data are of uniformly high quality down to our selection limit, our 16-hour exposures typically yielding errors of only \delta(dispersion)~10% for L* and fainter galaxies. By combining our measurements with data from the literature, we have 53 cluster galaxies with measured dispersions, and HST/ACS-derived sizes, colors and surface brightnesses. This sample is complete for the typical L* galaxy at z~1, unlike most previous z~1 cluster samples which are complete only for the massive cluster members (>1e11 M_sun). We find no evidence for a change in the tilt of the fundamental plane (FP). Nor do we find evidence for evolution in the slope of the color-dispersion relation and M/L_B-dispersion relations; measuring evolution at a fixed dispersion should minimize the impact of size evolution found in other work. The M/L_B at fixed dispersion evolves by \Delta log10 M/L_B=-0.50 +/- 0.03 between z=0.83 and z=0.02 or d(log10 M/L_B)=-0.60 +/- 0.04 dz, and we find \Delta (U-V)_z=-0.24 +/- 0.02 mag at fixed dispersion in the rest-frame, matching the expected evolution in M/L_B within 2.25 standard deviations. The implied formation redshift from both the color and M/L_B evolution is z*=2.0 +/- 0.2 +/- 0.3 (sys), during the epoch in which the cosmic star-formation activity peaked, with the systematic uncertainty showing the dependence of z* on the assumptions we make about the stellar populations. The lack of evolution in either the tilt of the FP or in the M/L- and color-dispersion relations imply that the formation epoch depends weakly on mass, ranging from z*=2.3 +1.3 -0.3 at 300 km/s to z*=1.7 +0.3 -0.2 at 160 km/s and implies that the IMF similarly varies slowly with galaxy mass.Comment: revised; typos corrected, references updated, and other cosmetic changes to meet ApJ format ApJ accepted, 22 pages in emulate ApJ format, 8 color figures, 1 b/w figur

The Kinematic Properties of BHB and RR Lyrae stars towards the Anticentre and the North Galactic Pole: The Transition between the Inner and the Outer Halo

We identify 51 blue horizontal branch (BHB) stars, 12 possible BHB stars and 58 RR Lyrae stars in Anticentre fields. Their selection does not depend on their kinematics. Light curves and ephemerides are given for 7 previously unknown RR Lyrae stars. All but 4 of the RR Lyrae stars are of Oosterhoff type I. Our selection criteria for BHB stars give results that agree with those used by Smith et al. (2010) and Ruhland et al. (2011). We use 5 methods to determine distances for the BHB stars and 3 methods for the RR Lyrae stars to get distances on a uniform scale. Absolute proper motions (largely derived from the GSCII and SDSS (DR7) databases) are given for these stars; radial velocities are given for 31 of the BHB stars and 37 of the RR Lyrae stars. Combining these data for BHB and RR Lyrae stars with those previously found in fields at the North Galactic Pole, we find that retrograde orbits dominate for galactocentric distances greater than 12.5 kpc. The majority of metal-poor stars in the solar neighbourhood are known to be concentrated in a Lperp vs. Lz angular momentum plot. We show that the ratio of the number of outliers to the number in the main concentration increases with galactocentric distance. The location of these outliers with Lperp and Lz shows that the halo BHB and RR Lyrae stars have more retrograde orbits and a more spherical distribution with increasing galactocentric distance. Six RR Lyrae stars are identified in the H99 group of outliers; the small spread in their [Fe/H] suggests that they could have come from a single globular cluster. Another group of outliers contains two pairs of RR Lyrae stars; the stars in each pair have similar properties.Comment: 40 pages, 19 figures, to be published in MNRA

The second data release of the INT Photometric Ha Survey of the Northern Galactic Plane (IPHAS DR2)

The INT/WFC Photometric Hα Survey of the Northern Galactic Plane (IPHAS) is a 1800 deg2 imaging survey covering Galactic latitudes |b| < 5° and longitudes ℓ = 30°–215° in the r, i, and Hα filters using the Wide Field Camera (WFC) on the 2.5-m Isaac Newton Telescope (INT) in La Palma. We present the first quality-controlled and globally calibrated source catalogue derived from the survey, providing single-epoch photometry for 219 million unique sources across 92 per cent of the footprint. The observations were carried out between 2003 and 2012 at a median seeing of 1.1 arcsec (sampled at 0.33 arcsec pixel−1) and to a mean 5σ depth of 21.2 (r), 20.0 (i), and 20.3 (Hα) in the Vega magnitude system. We explain the data reduction and quality control procedures, describe and test the global re-calibration, and detail the construction of the new catalogue. We show that the new calibration is accurate to 0.03 mag (root mean square) and recommend a series of quality criteria to select accurate data from the catalogue. Finally, we demonstrate the ability of the catalogue's unique (r − Hα, r − i) diagram to (i) characterize stellar populations and extinction regimes towards different Galactic sightlines and (ii) select and quantify Hα emission-line objects. IPHAS is the first survey to offer comprehensive CCD photometry of point sources across the Galactic plane at visible wavelengths, providing the much-needed counterpart to recent infrared surveys

Hypoxic Environment and Paired Hierarchical 3D and 2D Models of Pediatric H3.3-Mutated Gliomas Recreate the Patient Tumor Complexity.

BACKGROUND:Pediatric high-grade gliomas (pHGGs) are facing a very dismal prognosis and representative pre-clinical models are needed for new treatment strategies. Here, we examined the relevance of collecting functional, genomic, and metabolomics data to validate patient-derived models in a hypoxic microenvironment. METHODS:From our biobank of pediatric brain tumor-derived models, we selected 11 pHGGs driven by the histone H3.3K28M mutation. We compared the features of four patient tumors to their paired cell lines and mouse xenografts using NGS (next generation sequencing), aCGH (array comparative genomic hybridization), RNA sequencing, WES (whole exome sequencing), immunocytochemistry, and HRMAS (high resolution magic angle spinning) spectroscopy. We developed a multicellular in vitro model of cell migration to mimic the brain hypoxic microenvironment. The live cell technology Incucyte© was used to assess drug responsiveness in variable oxygen conditions. RESULTS:The concurrent 2D and 3D cultures generated from the same tumor sample exhibited divergent but complementary features, recreating the patient intra-tumor complexity. Genomic and metabolomic data described the metabolic changes during pHGG progression and supported hypoxia as an important key to preserve the tumor metabolism in vitro and cell dissemination present in patients. The neurosphere features preserved tumor development and sensitivity to treatment. CONCLUSION:We proposed a novel multistep work for the development and validation of patient-derived models, considering the immature and differentiated content and the tumor microenvironment of pHGGs

Pragmatic controlled clinical trials in primary care: the struggle between external and internal validity

BACKGROUND: Controlled clinical trials of health care interventions are either explanatory or pragmatic. Explanatory trials test whether an intervention is efficacious; that is, whether it can have a beneficial effect in an ideal situation. Pragmatic trials measure effectiveness; they measure the degree of beneficial effect in real clinical practice. In pragmatic trials, a balance between external validity (generalizability of the results) and internal validity (reliability or accuracy of the results) needs to be achieved. The explanatory trial seeks to maximize the internal validity by assuring rigorous control of all variables other than the intervention. The pragmatic trial seeks to maximize external validity to ensure that the results can be generalized. However the danger of pragmatic trials is that internal validity may be overly compromised in the effort to ensure generalizability. We are conducting two pragmatic randomized controlled trials on interventions in the management of hypertension in primary care. We describe the design of the trials and the steps taken to deal with the competing demands of external and internal validity. DISCUSSION: External validity is maximized by having few exclusion criteria and by allowing flexibility in the interpretation of the intervention and in management decisions. Internal validity is maximized by decreasing contamination bias through cluster randomization, and decreasing observer and assessment bias, in these non-blinded trials, through baseline data collection prior to randomization, automating the outcomes assessment with 24 hour ambulatory blood pressure monitors, and blinding the data analysis. SUMMARY: Clinical trials conducted in community practices present investigators with difficult methodological choices related to maintaining a balance between internal validity (reliability of the results) and external validity (generalizability). The attempt to achieve methodological purity can result in clinically meaningless results, while attempting to achieve full generalizability can result in invalid and unreliable results. Achieving a creative tension between the two is crucial

Nano-analyses of wear particles from metal-on-metal and non-metal-on-metal dual modular neck hip arthroplasty

Increased failure rates due to metallic wear particle-associated adverse local tissue reactions (ALTR) is a significant clinical problem in resurfacing and total hip arthroplasty. Retrieved periprosthetic tissue of 53 cases with corrosion/conventional metallic wear particles from 285 revision operations for ALTR was selected for nano-analyses. Three major classes of hip implants associated with ALTR, metal-on-metal hip resurfacing arthroplasty (MoM HRA) and large head total hip replacement (MoM LHTHA) and non-metal-on-metal dual modular neck total hip replacement (Non-MoM DMNTHA) were included. The size, shape, distribution, element composition, and crystal structure of the metal particles were analyzed by conventional histological examination and electron microscopy with analytic tools of 2D X-ray energy dispersive spectrometry and X-ray diffraction. Distinct differences in size, shape, and element composition of the metallic particles were detected in each implant class which correlate with the histological features of severity of ALTR and variability in implant performance