Lipid-induced hepatocyte-derived extracellular vesicles regulate hepatic stellate cell via microRNAs targeting PPAR-γ.

Background&aimsHepatic stellate cells (HSCs) play a key role in liver fibrosis in various chronic liver disorders including nonalcoholic fatty liver disease (NAFLD). The development of liver fibrosis requires a phenotypic switch from quiescent to activated HSCs. The triggers for HSCs activation in NAFLD remain poorly understood. We investigated the role and molecular mechanism of extracellular vesicles (EVs) released by hepatocytes during lipotoxicity in modulation of HSC phenotype.MethodsEVs were isolated from fat-laden hepatocytes by differential centrifugation and incubated with HSCs. EV internalization and HSCs activation, migration and proliferation were assessed. Loss- and gain-of-functions studies were performed to explore the potential role of PPAR-γ-targeting miRNAs carried by EVs into HSC.ResultsHepatocyte-derived EVs released during lipotoxicity are efficiently internalized by HSCs resulting in their activation, as shown by marked up-regulation of pro-fibrogenic genes (Collagen-I, α-SMA and TIMP-2), proliferation, chemotaxis and wound healing responses. These changes were associated with miRNAs shuttled by EVs and suppression of PPAR-γ expression in HSC. Hepatocyte-derived EVs miRNA content included various miRNAs that are known inhibitors of PPAR-γ expression with miR-128-3p being the most effectively transferred. Furthermore loss- and gain-of-function studies identified miR-128-3p as a central modulator of the effects of EVs on PPAR-γ inhibition and HSC activation.ConclusionOur findings demonstrate a link between fat-laden hepatocyte-derived EVs and liver fibrosis and have potential implications for the development of novel anti-fibrotic targets for NAFLD and other fibrotic diseases

The inverse-Compton ghost HDF 130 and the giant radio galaxy 6C 0905+3955: matching an analytic model for double radio source evolution

We present new GMRT observations of HDF 130, an inverse-Compton (IC) ghost of a giant radio source that is no longer being powered by jets. We compare the properties of HDF 130 with the new and important constraint of the upper limit of the radio flux density at 240 MHz to an analytic model. We learn what values of physical parameters in the model for the dynamics and evolution of the radio luminosity and X-ray luminosity (due to IC scattering of the cosmic microwave background (CMB)) of a Fanaroff-Riley II (FR II) source are able to describe a source with features (lobe length, axial ratio, X-ray luminosity, photon index and upper limit of radio luminosity) similar to the observations. HDF 130 is found to agree with the interpretation that it is an IC ghost of a powerful double-lobed radio source, and we are observing it at least a few Myr after jet activity (which lasted 5--100 Myr) has ceased. The minimum Lorentz factor of injected particles into the lobes from the hotspot is preferred to be $\gamma\sim10^3$ for the model to describe the observed quantities well, assuming that the magnetic energy density, electron energy density, and lobe pressure at time of injection into the lobe are linked by constant factors according to a minimum energy argument, so that the minimum Lorentz factor is constrained by the lobe pressure. We also apply the model to match the features of 6C 0905+3955, a classical double FR II galaxy thought to have a low-energy cutoff of $\gamma\sim10^4$ in the hotspot due to a lack of hotspot inverse-Compton X-ray emission. The models suggest that the low-energy cutoff in the hotspots of 6C 0905+3955 is $\gamma\gtrsim 10^3$, just slightly above the particles required for X-ray emission.Comment: 9 pages, 3 figure

Action research in physical education: focusing beyond myself through cooperative learning

This paper reports on the pedagogical changes that I experienced as a teacher engaged in an action research project in which I designed and implemented an indirect, developmentally appropriate and child‐centred approach to my teaching. There have been repeated calls to expunge – or at least rationalise – the use of traditional, teacher‐led practice in physical education. Yet despite the advocacy of many leading academics there is little evidence that such a change of approach is occurring. In my role as teacher‐as‐researcher I sought to implement a new pedagogical approach, in the form of cooperative learning, and bring about a positive change in the form of enhanced pupil learning. Data collection included a reflective journal, post‐teaching reflective analysis, pupil questionnaires, student interviews, document analysis, and non‐participant observations. The research team analysed the data using inductive analysis and constant comparison. Six themes emerged from the data: teaching and learning, reflections on cooperation, performance, time, teacher change, and social interaction. The paper argues that cooperative learning allowed me to place social and academic learning goals on an even footing, which in turn placed a focus on pupils’ understanding and improvement of skills in athletics alongside their interpersonal development

How does reviewing the evidence change veterinary surgeons' beliefs regarding the treatment of ovine footrot? A quantitative and qualitative study

Footrot is a widespread, infectious cause of lameness in sheep, with major economic and welfare costs. The aims of this research were: (i) to quantify how veterinary surgeons’ beliefs regarding the efficacy of two treatments for footrot changed following a review of the evidence (ii) to obtain a consensus opinion following group discussions (iii) to capture complementary qualitative data to place their beliefs within a broader clinical context. Grounded in a Bayesian statistical framework, probabilistic elicitation (roulette method) was used to quantify the beliefs of eleven veterinary surgeons during two one-day workshops. There was considerable heterogeneity in veterinary surgeons’ beliefs before they listened to a review of the evidence. After hearing the evidence, seven participants quantifiably changed their beliefs. In particular, two participants who initially believed that foot trimming with topical oxytetracycline was the better treatment, changed to entirely favour systemic and topical oxytetracycline instead. The results suggest that a substantial amount of the variation in beliefs related to differences in veterinary surgeons’ knowledge of the evidence. Although considerable differences in opinion still remained after the evidence review, with several participants having non-overlapping 95% credible intervals, both groups did achieve a consensus opinion. Two key findings from the qualitative data were: (i) veterinary surgeons believed that farmers are unlikely to actively seek advice on lameness, suggesting a proactive veterinary approach is required (ii) more attention could be given to improving the way in which veterinary advice is delivered to farmers. In summary this study has: (i) demonstrated a practical method for probabilistically quantifying how veterinary surgeons’ beliefs change (ii) revealed that the evidence that currently exists is capable of changing veterinary opinion (iii) suggested that improved transfer of research knowledge into veterinary practice is needed (iv) identified some potential obstacles to the implementation of veterinary advice by farmers

Donor insulin use predicts beta‐cell function after islet transplantation

Insulin is routinely used to manage hyperglycaemia in organ donors and during the peri-transplant period in islet transplant recipients. However, it is unknown whether donor insulin use (DIU) predicts beta-cell dysfunction after islet transplantation. We reviewed data from the UK Transplant Registry and the UK Islet Transplant Consortium; all first-time transplants during 2008-2016 were included. Linear regression models determined associations between DIU, median and coefficient of variation (CV) peri-transplant glucose levels and 3-month islet graft function. In 91 islet cell transplant recipients, DIU was associated with lower islet function assessed by BETA-2 scores (β [SE] -3.5 [1.5], P = .02), higher 3-month post-transplant HbA1c levels (5.4 [2.6] mmol/mol, P = .04) and lower fasting C-peptide levels (−107.9 [46.1] pmol/l, P = .02). Glucose at 10 512 time points was recorded during the first 5 days peri-transplant: the median (IQR) daily glucose level was 7.9 (7.0-8.9) mmol/L and glucose CV was 28% (21%-35%). Neither median glucose levels nor glucose CV predicted outcomes post-transplantation. Data on DIU predicts beta-cell dysfunction 3 months after islet transplantation and could help improve donor selection and transplant outcomes

High Affinity for Farnesyltransferase and Alternative Prenylation Contribute Individually to K-Ras4B Resistance to Farnesyltransferase Inhibitors

Farnesyltransferase inhibitors (FTIs) block Ras farnesylation, subcellular localization and activity, and inhibit the growth of Ras-transformed cells. Although FTIs are ineffective against K-Ras4B, the Ras isoform most commonly mutated in human cancers, they can inhibit the growth of tumors containing oncogenic K-Ras4B, implicating other farnesylated proteins or suggesting distinct functions for farnesylated and for geranylgeranylated K-Ras, which is generated when farnesyltransferase is inhibited. In addition to bypassing FTI blockade through geranylgeranylation, K-Ras4B resistance to FTIs may also result from its higher affinity for farnesyltransferase. Using chimeric Ras proteins containing all combinations of Ras background, CAAX motif, and K-Ras polybasic domain, we show that either a polybasic domain or an alternatively prenylated CAAX renders Ras prenylation, Ras-induced Elk-1 activation, and anchorage-independent cell growth FTI-resistant. The polybasic domain alone increases the affinity of Ras for farnesyltransferase, implying independent roles for each K-Ras4B sequence element in FTI resistance. Using microarray analysis and colony formation assays, we confirm that K-Ras function is independent of the identity of the prenyl group and, therefore, that FTI inhibition of K-Ras transformed cells is likely to be independent of K-Ras inhibition. Our results imply that relevant FTI targets will lack both polybasic and potentially geranylgeranylated methionine-CAAX motifs