46 research outputs found

    Mechanical tests and definition of new indexes of grape berry firmness. Evolution of berry skin hardness during alcoholic fermentation

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    The mechanical strength or firmness of a fruit is considered an important parameter to characterise its state of ripeness or conservation, as well as other parameters such as sugar level or color. The mechanical hardness of grapes influences the integrity and sanitary quality of the harvest. In this study, the mechanical characteristics of grapevine berries were studied at harvest time in order to determine their rheological properties (firmness and hardness of the berry skin) during alcoholic fermentation. Special indexes were defined measuring the energy needed to crush the berries to 50 % of their initial diameter, and applied successively to two different varieties. The entire berry firmness and the skin hardness were both different. Mechanical indexes linked to grape firmness were defined. Using these indexes, a significant effect on the firmness behavior due to variety was recorded: the skin of 'Grenache Noir' was found firmer and harder than 'Carignan Noir'. Furthermore, during the alcoholic fermentation, no change in skin hardness was observed for both varieties, despite changes in the composition of the must. These results give new information on mechanical properties of berries and could be used as an aid in the winemaking process. Indeed, they would probably help the winemaker to better choose the type of fermentation and maceration adapted to his grapes according to the type of wine he wishes to produce

    Flotillins Interact with PSGL-1 in Neutrophils and, upon Stimulation, Rapidly Organize into Membrane Domains Subsequently Accumulating in the Uropod

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    BACKGROUND: Neutrophils polarize and migrate in response to chemokines. Different types of membrane microdomains (rafts) have been postulated to be present in rear and front of polarized leukocytes and disruption of rafts by cholesterol sequestration prevents leukocyte polarization. Reggie/flotillin-1 and -2 are two highly homologous proteins that are ubiquitously enriched in detergent resistant membranes and are thought to shape membrane microdomains by forming homo- and hetero-oligomers. It was the goal of this study to investigate dynamic membrane microdomain reorganization during neutrophil activation. METHODOLOGY/PRINCIPAL FINDINGS: We show now, using immunofluorescence staining and co-immunoprecipitation, that endogenous flotillin-1 and -2 colocalize and associate in resting spherical and polarized primary neutrophils. Flotillins redistribute very early after chemoattractant stimulation, and form distinct caps in more than 90% of the neutrophils. At later time points flotillins accumulate in the uropod of polarized cells. Chemotactic peptide-induced redistribution and capping of flotillins requires integrity and dynamics of the actin cytoskeleton, but does not involve Rho-kinase dependent signaling related to formation of the uropod. Both flotillin isoforms are involved in the formation of this membrane domain, as uropod location of exogenously expressed flotillins is dramatically enhanced by co-overexpression of tagged flotillin-1 and -2 in differentiated HL-60 cells as compared to cells expressing only one tagged isoform. Flotillin-1 and -2 associate with P-selectin glycoprotein ligand 1 (PSGL-1) in resting and in stimulated neutrophils as shown by colocalization and co-immunoprecipitation. Neutrophils isolated from PSGL-1-deficient mice exhibit flotillin caps to the same extent as cells isolated from wild type animals, implying that PSGL-1 is not required for the formation of the flotillin caps. Finally we show that stimulus-dependent redistribution of other uropod-located proteins, CD43 and ezrin/radixin/moesin, occurs much slower than that of flotillins and PSGL-1. CONCLUSIONS/SIGNIFICANCE: These results suggest that flotillin-rich actin-dependent membrane microdomains are importantly involved in neutrophil uropod formation and/or stabilization and organize uropod localization of PSGL-1

    Thousands of Rab GTPases for the Cell Biologist

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    Rab proteins are small GTPases that act as essential regulators of vesicular trafficking. 44 subfamilies are known in humans, performing specific sets of functions at distinct subcellular localisations and tissues. Rab function is conserved even amongst distant orthologs. Hence, the annotation of Rabs yields functional predictions about the cell biology of trafficking. So far, annotating Rabs has been a laborious manual task not feasible for current and future genomic output of deep sequencing technologies. We developed, validated and benchmarked the Rabifier, an automated bioinformatic pipeline for the identification and classification of Rabs, which achieves up to 90% classification accuracy. We cataloged roughly 8.000 Rabs from 247 genomes covering the entire eukaryotic tree. The full Rab database and a web tool implementing the pipeline are publicly available at www.RabDB.org. For the first time, we describe and analyse the evolution of Rabs in a dataset covering the whole eukaryotic phylogeny. We found a highly dynamic family undergoing frequent taxon-specific expansions and losses. We dated the origin of human subfamilies using phylogenetic profiling, which enlarged the Rab repertoire of the Last Eukaryotic Common Ancestor with Rab14, 32 and RabL4. Furthermore, a detailed analysis of the Choanoflagellate Monosiga brevicollis Rab family pinpointed the changes that accompanied the emergence of Metazoan multicellularity, mainly an important expansion and specialisation of the secretory pathway. Lastly, we experimentally establish tissue specificity in expression of mouse Rabs and show that neo-functionalisation best explains the emergence of new human Rab subfamilies. With the Rabifier and RabDB, we provide tools that easily allows non-bioinformaticians to integrate thousands of Rabs in their analyses. RabDB is designed to enable the cell biology community to keep pace with the increasing number of fully-sequenced genomes and change the scale at which we perform comparative analysis in cell biology

    Lipid raft adhesion receptors and Syk regulate selectin-dependent rolling under flow conditions

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    Selectins and their ligand P-selectin glycoprotein ligand-1 (PSGL-1) mediate leukocyte rolling along inflamed vessels. Cell rolling is modulated by selectin interactions with their ligands and by topographic requirements including L-selectin and PSGL-1 clustering on tips of leukocyte microvilli. Lipid rafts are cell membrane microdomains reported to function as signaling platforms. Here, we show that disruption of leukocyte lipid rafts with cholesterol chelating agents depleted raft-associated PSGL-1 and L-selectin and strongly reduced L-, P-, and E-selectin-dependent rolling. Cholesterol repletion reversed inhibition of cell rolling. Importantly, leukocyte rolling on P-selectin induced the recruitment of spleen tyrosine kinase (Syk), a tyrosine kinase associated to lipid raft PSGL-1. Furthermore, inhibition of Syk activity or expression, with pharmacologic inhibitors or by RNA interference, strongly reduced leukocyte rolling on P-selectin, but not on E-selectin or PSGL-1. These observations identify novel regulatory mechanisms of leukocyte rolling on selectins with a strong dependency on lipid raft integrity and Syk activity

    Interleukin-4 induces functional cell-surface expression of CXCR4 on human T cells

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    Bases physiopathologiques en hématologie générale : un aide-mémoire d'hématologie

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    La disciplina contable como ciencia no ha sido ajena a los cambios que generan las crisis en los ámbitos económico, político y social. El presente trabajo, que forma parte de un proyecto de investigación, hace un recorrido histórico abordando la evolución de la contabilidad de gestión a partir de la investigación y análisis de bibliografía nacional y extranjera. Se plantea como objetivos de trabajo los siguientes: - Caracterizar la evolución histórica de la contabilidad de gestión - Relacionar el surgimiento de nuevas herramientas de gestión con la evolución de los paradigmas económicos. - Plantear la utilización del ‘Cuadro de mando Integral’ (Balanced Scorecard -BSC) como herramienta para incorporar la dimensión social y ambiental en la gestión empresaria.Centro de Estudios en Contabilidad Internacional (CECIN
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