Evaluating the cellular consequences of ACLY inhibition in primary hepatocytes and pancreatic islets

Motivation: Metabolic diseases and their pandemic proportions cause a significant loss in quality and expectancy of life in around 25% of the population. Therefore, the development of more efficient therapies should be a clinical priority. This project aims to evaluate the cellular consequences of a chronic Ac-CoA cytosolic pool reduction induced by the inhibition of ATP-citrate lyase (ACLY), core enzyme in metabolism and major source of Ac-CoA within the cytoplasm. Ac-Coa is a central molecule in many metabolic pathways involved in de novo lipogenesis, cholesterol production and protein acetylation.Methods: Most of the assays were performed on mouse primary cultured hepatocytes and islets of Langerhans. Hepatocytes were obtained by perfusing the mouse liver with collagenase and isolated by filtering and differential centrifugation. Islets of Langerhans were obtained by perfusing the mouse pancreas with collagenase and were isolated by handpicking. Cells were treated with 0, 10, 30, 50 and 100 μM of SB-204990 (SB), an ACLY inhibitor with a Ki of 1 μM. Metabolic cell activity, cell death and urea secretion were measured. Radioactive metabolite tracing was performed to assess ACLY-dependent metabolic functions. Proteins of treated hepatocytes were extracted to perform western blots.Results: In primary hepatocytes, SB neither affected metabolic activity nor urea production when concentration was <50 μM SB. However, exposure to this drug resulted in a significant increase in cell toxicity at ≥30 μM SB. We detected a dose-dependent decrease in liponeogenesis upon treatment. β-oxidation of fatty acids was affected by SB following a hormetic effect, with a rise at 10 μM and a reduction at 50 and 100 μM. The GAPDH protein expression was higher at 10 μM SB, suggesting a modulation of glycolysis/gluconeogenesis. LC3B protein expression patterns indicated autophagy was reduced at 10 μM SB and incremented at 100 μM SB. In primary islets, SB did not impair metabolic activity, while cell death, determined by ELISA, showed a reduction at 30 μM and no effects at other concentrations.Conclusions: The effects of ACLY inhibition showed that this enzyme plays a key role in the cellular homeostasis of primary hepatocytes. Fatty acids catabolism and glucose utilization relies on the presence of functional ACLY. In islets, SB revealed to be non toxic. Our results suggest that ACLY may represent a promising target for the treatment of metabolc disorders

Fermion Masses in Emergent Electroweak Symmetry Breaking

We consider the generation of fermion masses in an emergent model of electroweak symmetry breaking with composite $W,Z$ gauge bosons. A universal bulk fermion profile in a warped extra dimension is used for all fermion flavors. Electroweak symmetry is broken at the UV (or Planck) scale where boundary mass terms are added to generate the fermion flavor structure. This leads to flavor-dependent nonuniversality in the gauge couplings. The effects are suppressed for the light fermion generations but are enhanced for the top quark where the $Zt{\bar t}$ and $Wt{\bar b}$ couplings can deviate at the $10-20$ level in the minimal setup. By the AdS/CFT correspondence our model implies that electroweak symmetry is not a fundamental gauge symmetry. Instead the Standard Model with massive fermions and $W,Z$ gauge bosons is an effective chiral Lagrangian for some underlying confining strong dynamics at the TeV scale, where mass is generated without a Higgs mechanism.Comment: modified discussion in Sec 3.1, version published in JHE

The Close AGN Reference Survey (CARS) A massive multi-phase outflow impacting the edge-on galaxy HE 1353-1917

Open Access article, published by EDP Sciences, under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Open Access funding provided by Max Planck Society.Context. Galaxy-wide outflows driven by star formation and/or an active galactic nucleus (AGN) are thought to play a crucial rule in the evolution of galaxies and the metal enrichment of the inter-galactic medium. Direct measurements of these processes are still scarce and new observations are needed to reveal the nature of outflows in the majority of the galaxy population. Aims. We combine extensive, spatially-resolved, multi-wavelength observations, taken as part of the Close AGN Reference Survey (CARS), for the edge-on disc galaxy HE 1353-1917 in order to characterise the impact of the AGN on its host galaxy via outflows and radiation. Methods. Multi-color broad-band photometry was combined with spatially-resolved optical, near-infrared (NIR) and sub-mm and radio observations taken with the Multi-Unit Spectroscopy Explorer (MUSE), the Near-infrared Integral Field Spectrometer (NIFS), the Atacama Large Millimeter Array (ALMA), and the Karl G. Jansky Very Large Array (VLA) to map the physical properties and kinematics of the multi-phase interstellar medium. Results. We detect a biconical extended narrow-line region ionised by the luminous AGN orientated nearly parallel to the galaxy disc, extending out to at least 25 kpc. The extra-planar gas originates from galactic fountains initiated by star formation processes in the disc, rather than an AGN outflow, as shown by the kinematics and the metallicity of the gas. Nevertheless, a fast, multi-phase, AGN-driven outflow with speeds up to 1000 km s(-1) is detected close to the nucleus at 1 kpc distance. A radio jet, in connection with the AGN radiation field, is likely responsible for driving the outflow as confirmed by the energetics and the spatial alignment of the jet and multi-phase outflow. Evidence for negative AGN feedback suppressing the star formation rate (SFR) is mild and restricted to the central kpc. But while any SFR suppression must have happened recently, the outflow has the potential to greatly impact the future evolution of the galaxy disc due to its geometrical orientation. Conclusions.. Our observations reveal that low-power radio jets can play a major role in driving fast, multi-phase, galaxy-scale outflows even in radio-quiet AGN. Since the outflow energetics for HE 1353-1917 are consistent with literature, scaling relation of AGN-driven outflows the contribution of radio jets as the driving mechanisms still needs to be systematically explored.© B. Husemann et al. 2019We thank the referee for providing very valuable comments, which significantly improved the quality of the manuscript. MK acknowledges support from DLR grant 50OR1802. GRT acknowledges support from the NASA through Einstein Postdoctoral Fellowship Award Number PF-150128, issued by the Chandra X-ray Observatory Center, which is operated by the Smithsonian Astrophysical Observatory for and on behalf of NASA under contract NAS8-03060. MG is supported by the Lyman Spitzer Jr. Fellowship (Princeton University) and by NASA Chandra grants GO7-18121X/GO8-19104X. SMC acknowledges support from the Australian Research Council (DP190102714). We thank Alex Markowitz for helpful discussions on the RGS data in the context of warm absorbers. The work of SAB, CPO and MS was supported by a generous grant from the Natural Sciences and Engineering Research Council of Canada. Based on observations collected at the European Organization for Astronomical Research in the Southern Hemisphere under ESO programme 095. B-0015(A). Based on observations obtained at the Gemini Observatory, which is operated by the Association of Universities for Research in Astronomy, Inc., under a cooperative agreement with the NSF on behalf of the Gemini partnership: the National Science Foundation (United States), National Research Council (Canada), CONICYT (Chile), Ministerio de Ciencia, Tecnologia e Innovacion Productiva (Argentina), Ministerio da Ciencia, Tecnologia e Inovacao (Brazil), and Korea Astronomy and Space Science Institute (Republic of Korea). Based on observations obtained at the Southern Astrophysical Research (SOAR) telescope, which is a joint project of the Ministerio da Ciencia, Tecnologia, Inovacoes e Comunicacoes (MCTIC) do Brasil, the U.S. National Optical Astronomy Observatory (NOAO), the University of North Carolina at Chapel Hill (UNC), and Michigan State University (MSU). Based on observations collected at the German-Spanish Astronomical Center, Calar Alto, jointly operated by the Max-Planck-Institut fur Astronomie Heidelberg and the Instituto de Astrofiica de Andaluci (CSIC). This paper makes use of the following ALMA data: ADS/JAO.ALMA#2016.1.00952. S. ALMA is a partnership of ESO (representing its member states), NSF (USA) and NINS (Japan), together with NRC (Canada), MOST and ASIAA (Taiwan), and KASI (Republic of Korea), in cooperation with the Republic of Chile. The Joint ALMA Observatory is operated by ESO, AUI/NRAO and NAOJ. This work is based on observations obtained with XMM-Newton, an ESA science mission with instruments and contributions directly funded by ESA Member States and NASA. The VLA is operated by the National Radio Astronomy Observatory, a facility of the National Science Foundation operated under cooperative agreement by Associated Universities, Inc. Herschel is an ESA space observatory with science instruments provided by European-led Principal Investigator consortia and with important participation from NASA. This publication makes use of data products from the Wide-field Infrared Survey Explorer, which is a joint project of the University of California, Los Angeles, and the Jet Propulsion Laboratory/California Institute of Technology, funded by the National Aeronautics and Space Administration. This research has made use of the NASA/IPAC Infrared Science Archive, which is operated by the Jet Propulsion Laboratory, California Institute of Technology, under contract with the National Aeronautics and Space Administration. The Pan-STARRS1 Surveys (PS1) and the PS1 public science archive have been made possible through contributions by the Institute for Astronomy, the University of Hawaii, the Pan-STARRS Project Office, the Max-Planck Society and its participating institutes, the Max Planck Institute for Astronomy, Heidelberg and the Max Planck Institute for Extraterrestrial Physics, Garching, The Johns Hopkins University, Durham University, the University of Edinburgh, the Queen's University Belfast, the Harvard-Smithsonian Center for Astrophysics, the Las Cumbres Observatory Global Telescope Network Incorporated, the National Central University of Taiwan, the Space Telescope Science Institute, the National Aeronautics and Space Administration under Grant No. NNX08AR22G issued through the Planetary Science Division of the NASA Science Mission Directorate, the National Science Foundation Grant No. AST-1238877, the University of Maryland, Eotvos Lorand University (ELTE), the Los Alamos National Laboratory, and the Gordon and Betty Moore Foundation. This work is based in part on observations made with the Galaxy Evolution Explorer (GALEX). GALEX is a NASA Small Explorer, whose mission was developed in cooperation with the Centre National d'Etudes Spatiales (CNES) of France and the Korean Ministry of Science and Technology. GALEX is operated for NASA by the California Institute of Technology under NASA contract NAS5-98034

Transcriptome profiling of grapevine seedless segregants during berry development reveals candidate genes associated with berry weight

Indexación: Web of Science; PubMedBackground Berry size is considered as one of the main selection criteria in table grape breeding programs. However, this is a quantitative and polygenic trait, and its genetic determination is still poorly understood. Considering its economic importance, it is relevant to determine its genetic architecture and elucidate the mechanisms involved in its expression. To approach this issue, an RNA-Seq experiment based on Illumina platform was performed (14 libraries), including seedless segregants with contrasting phenotypes for berry weight at fruit setting (FST) and 6–8 mm berries (B68) phenological stages. Results A group of 526 differentially expressed (DE) genes were identified, by comparing seedless segregants with contrasting phenotypes for berry weight: 101 genes from the FST stage and 463 from the B68 stage. Also, we integrated differential expression, principal components analysis (PCA), correlations and network co-expression analyses to characterize the transcriptome profiling observed in segregants with contrasting phenotypes for berry weight. After this, 68 DE genes were selected as candidate genes, and seven candidate genes were validated by real time-PCR, confirming their expression profiles. Conclusions We have carried out the first transcriptome analysis focused on table grape seedless segregants with contrasting phenotypes for berry weight. Our findings contributed to the understanding of the mechanisms involved in berry weight determination. Also, this comparative transcriptome profiling revealed candidate genes for berry weight which could be evaluated as selection tools in table grape breeding programs.http://bmcplantbiol.biomedcentral.com/articles/10.1186/s12870-016-0789-

Biotechnological production and application of fructooligosaccharides

Currently, prebiotics are all carbohydrates of relatively short chain length. An important group is the fructooligosaccharides, which are a special kind of prebiotics associated to their selective stimulation of the activity of certain groups of colonic bacteria that have a positive and beneficial effect on intestinal microbiota, reducing incidence of gastrointestinal infections, respiratory and also possessing a recognized bifidogenic effect. Traditionally, these prebiotic compounds have been obtained through extraction processes from some plants, as well as through enzymatic hydrolysis of sucrose. However, different fermentative methods have also been proposed for the production of fructooligosaccharides, such as solid-state fermentation utilizing various agroindustrial by-products. By optimizing the culture parameters, fructooligosaccharides yields and productivity can be improved. The use of immobilized enzymes and cells has also been proposed as being an effective and economic method for large-scale production of fructooligosaccharides. This paper is an overview on the results of recent studies on fructooligosacharides biosynthesis, physicochemical properties, sources, biotechnological production and applications.The authors thank the National Council of Science and Technology of Mexico (CONACYT) for funding this study. D. A. Flores-Maltos thank the CONACYT for the financial support provided for her postgraduate studies in the Food Science and Technology Program, Universidad Autonoma de Coahuila, Mexico

The Cytotoxic Necrotizing Factor of Yersinia pseudotuberculosis (CNFy) is Carried on Extracellular Membrane Vesicles to Host Cells

In this study we show Yersinia pseudotuberculosis secretes membrane vesicles (MVs) that contain different proteins and virulence factors depending on the strain. Although MVs from Y. pseudotuberculosis YPIII and ATCC 29833 had many proteins in common (68.8% of all the proteins identified), those located in the outer membrane fraction differed significantly. For instance, the MVs from Y. pseudotuberculosis YPIII harbored numerous Yersinia outer proteins (Yops) while they were absent in the ATCC 29833 MVs. Another virulence factor found solely in the YPIII MVs was the cytotoxic necrotizing factor (CNFy), a toxin that leads to multinucleation of host cells. The ability of YPIII MVs to transport this toxin and its activity to host cells was verified using HeLa cells, which responded in a dose-dependent manner; nearly 70% of the culture was multinucleated after addition of 5 mu g/ml of the purified YPIII MVs. In contrast, less than 10% were multinucleated when the ATCC 29833 MVs were added. Semi-quantification of CNFy within the YPIII MVs found this toxin is present at concentrations of 5 -10 ng per mu g of total MV protein, a concentration that accounts for the cellular responses see

RT-GENE: Real-time eye gaze estimation in natural environments

In this work, we consider the problem of robust gaze estimation in natural environments. Large camera-to-subject distances and high variations in head pose and eye gaze angles are common in such environments. This leads to two main shortfalls in state-of-the-art methods for gaze estimation: hindered ground truth gaze annotation and diminished gaze estimation accuracy as image resolution decreases with distance. We first record a novel dataset of varied gaze and head pose images in a natural environment, addressing the issue of ground truth annotation by measuring head pose using a motion capture system and eye gaze using mobile eyetracking glasses. We apply semantic image inpainting to the area covered by the glasses to bridge the gap between training and testing images by removing the obtrusiveness of the glasses. We also present a new real-time algorithm involving appearance-based deep convolutional neural networks with increased capacity to cope with the diverse images in the new dataset. Experiments with this network architecture are conducted on a number of diverse eye-gaze datasets including our own, and in cross dataset evaluations. We demonstrate state-of-the-art performance in terms of estimation accuracy in all experiments, and the architecture performs well even on lower resolution images

About 1% of the breast and ovarian Spanish families testing negative for BRCA1 and BRCA2 are carriers of RAD51D pathogenic variants

RAD51D mutations have been recently identified in breast (BC) and ovarian cancer (OC) families. Although an etiological role in OC appears to be present, the association of RAD51D mutations and BC risk is more unclear. We aimed to determine the prevalence of germline RAD51D mutations in Spanish BC/OC families negative for BRCA1/BRCA2 mutations. We analyzed 842 index patients: 491 from BC/OC families, 171 BC families, 51 OC families and 129 patients without family history but with early-onset BC or OC or metachronous BC and OC. Mutation detection was performed with high-resolution melting, denaturing high-performance liquid chromatography or Sanger sequencing. Three mutations were found in four families with BC and OC cases (0.82%). Two were novel: c.1A>T (p.Met1?) and c.667+2_667+23del, leading to the exon 7 skipping and one previously described: c.674C>T (p.Arg232*). All were present in BC/OC families with only one OC. The c.667+2_667+23del cosegregated in the family with one early-onset BC and two bilateral BC cases. We also identified the c.629C>T (p.Ala210Val) variant, which was predicted in silico to be potentially pathogenic. About 1% of the BC and OC Spanish families negative for BRCA1/BRCA2 are carriers of RAD51D mutations. The presence of several BC mutation carriers, albeit in the context of familial OC, suggests an increased risk for BC, which should be taken into account in the follow-up and early detection measures. RAD51D testing should be considered in clinical setting for families with BC and OC, irrespective of the number of OC cases in the family

Study protocol of psychometric properties of the Spanish translation of a competence test in evidence based practice: The Fresno test

<p>Abstract</p> <p>Background</p> <p>There are few high-quality instruments for evaluating the effectiveness of Evidence-Based Practice (EBP) curricula with objective outcomes measures. The Fresno test is an instrument that evaluates most of EBP steps with a high reliability and validity in the English original version. The present study has the aims to translate the Fresno questionnaire into Spanish and its subsequent validation to ensure the equivalence of the Spanish version against the English original.</p> <p>Methods and design</p> <p>The questionnaire will be translated with the back translation technique and tested in Primary Care Teaching Units in Catalonia (PCTU). Participants will be: (a) tutors of Family Medicine residents (expert group); (b) Family Medicine residents in their second year of the Family Medicine training program (novice group), and (c) Family Medicine physicians (intermediate group). The questionnaire will be administered before and after an educational intervention. The educational intervention will be an interactive four half-day sessions designed to develop the knowledge and skills required to EBP. Responsiveness statistics used in the analysis will be the effect size, the standardised response mean and Guyatt's method. For internal consistency reliability, two measures will be used: corrected item-total correlations and Cronbach's alpha. Inter-rater reliability will be tested using Kappa coefficient for qualitative items and intra-class correlation coefficient for quantitative items and the overall score. Construct validity, item difficulty, item discrimination and feasibility will be determined.</p> <p>Discussion</p> <p>The validation of the Fresno questionnaire into different languages will enable the expansion of the questionnaire, as well as allowing comparison between countries and the evaluation of different teaching models.</p

The stellar and sub-stellar IMF of simple and composite populations

The current knowledge on the stellar IMF is documented. It appears to become top-heavy when the star-formation rate density surpasses about 0.1Msun/(yr pc^3) on a pc scale and it may become increasingly bottom-heavy with increasing metallicity and in increasingly massive early-type galaxies. It declines quite steeply below about 0.07Msun with brown dwarfs (BDs) and very low mass stars having their own IMF. The most massive star of mass mmax formed in an embedded cluster with stellar mass Mecl correlates strongly with Mecl being a result of gravitation-driven but resource-limited growth and fragmentation induced starvation. There is no convincing evidence whatsoever that massive stars do form in isolation. Various methods of discretising a stellar population are introduced: optimal sampling leads to a mass distribution that perfectly represents the exact form of the desired IMF and the mmax-to-Mecl relation, while random sampling results in statistical variations of the shape of the IMF. The observed mmax-to-Mecl correlation and the small spread of IMF power-law indices together suggest that optimally sampling the IMF may be the more realistic description of star formation than random sampling from a universal IMF with a constant upper mass limit. Composite populations on galaxy scales, which are formed from many pc scale star formation events, need to be described by the integrated galactic IMF. This IGIMF varies systematically from top-light to top-heavy in dependence of galaxy type and star formation rate, with dramatic implications for theories of galaxy formation and evolution.Comment: 167 pages, 37 figures, 3 tables, published in Stellar Systems and Galactic Structure, Vol.5, Springer. This revised version is consistent with the published version and includes additional references and minor additions to the text as well as a recomputed Table 1. ISBN 978-90-481-8817-