82 research outputs found

    Annexin-A5 assembled into two-dimensional arrays promotes cell membrane repair

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    Eukaryotic cells possess a universal repair machinery that ensures rapid resealing of plasma membrane disruptions. Before resealing, the torn membrane is submitted to considerable tension, which functions to expand the disruption. Here we show that annexin-A5 (AnxA5), a protein that self-assembles into two-dimensional (2D) arrays on membranes upon Ca2+ activation, promotes membrane repair. Compared with wild-type mouse perivascular cells, AnxA5-null cells exhibit a severe membrane repair defect. Membrane repair in AnxA5-null cells is rescued by addition of AnxA5, which binds exclusively to disrupted membrane areas. In contrast, an AnxA5 mutant that lacks the ability of forming 2D arrays is unable to promote membrane repair. We propose that AnxA5 participates in a previously unrecognized step of the membrane repair process: triggered by the local influx of Ca2+, AnxA5 proteins bind to torn membrane edges and form a 2D array, which prevents wound expansion and promotes membrane resealing

    Metabolic effects of elevated temperature on organic acid degradation in ripening <em>Vitis vinifera</em> fruit

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    First published online: September 1, 2014Berries of the cultivated grapevine Vitis vinifera are notably responsive to temperature, which can influence fruit quality and hence the future compatibility of varieties with their current growing regions. Organic acids represent a key component of fruit organoleptic quality and their content is significantly influenced by temperature. The objectives of this study were to (i) manipulate thermal regimes to realistically capture warming-driven reduction of malate content in Shiraz berries, and (ii) investigate the mechanisms behind temperature-sensitive malate loss and the potential downstream effects on berry metabolism. In the field we compared untreated controls at ambient temperature with longer and milder warming (2-4 °C differential for three weeks; Experiment 1) or shorter and more severe warming (4-6 °C differential for 11 days; Experiment 2). We complemented field trials with control (25/15 °C) and elevated (35/20 °C) day/night temperature controlled-environment trials using potted vines (Experiment 3). Elevating maximum temperatures (4-10 °C above controls) during pre-véraison stages led to higher malate content, particularly with warmer nights. Heating at véraison and ripening stages reduced malate content, consistent with effects typically seen in warm vintages. However, when minimum temperatures were also raised by 4-6 °C, malate content was not reduced, suggesting that the regulation of malate metabolism differs during the day and night. Increased NAD-dependent malic enzyme activity and decreased phosphoenolpyruvate carboxylase and pyruvate kinase activities, as well as the accumulation of various amino acids and γ-aminobutyric acid, suggest enhanced anaplerotic capacity of the TCA cycle and a need for coping with decreased cytosolic pH in heated fruit.C. Sweetman, V. O. Sadras, R. D. Hancock, K. L. Soole and C. M. For

    Proteomic Analysis of Grape Berry Cell Cultures Reveals that Developmentally Regulated Ripening Related Processes Can Be Studied Using Cultured Cells

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    The original publication is available at http:/www.plosone.orgBackground: This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first. Methodology/Principal Findings: In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions. Conclusions: The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening. © 2011 Sharathchandra et al.Publishers' Versio

    Guidance on a better integration of aquaculture, fisheries, and other activities in the coastal zone: from tools to practical examples

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    This guidance document provides a comprehensive assessment of the conflicts and synergies between fisheries, aquaculture and other activities in the coastal zone in six COEXIST case study areas. It forms deliverable D5.2 of the COEXIST project and synthesises deliverable D5.1, which provides a more detailed description of the methods used and results. This document also accounts for the views and expectations of stakeholders that were raised at the COEXIST stakeholder workshop held in Bergen, Norway, parallel to the ICES (International Council for the Exploration of the Sea) Annual Science Conference 2012. Over 30 stakeholders representing a variety of sectors, including aquaculture, fisheries, coastal zone management, tourism and energy, as well as 20 members from the COEXIST project and ICES representatives, attended this event. The stakeholders and COEXIST members were from Denmark, Finland, France, Germany, Ireland, Italy, Norway, Portugal, Spain, the Netherlands and the United Kingdom. The workshop aims were firstly to communicate the COEXIST project results and progress to stakeholders and the second major aim was to receive stakeholder feedback on the development of best practice guidance for spatial planning to integrate fisheries, aquaculture and further demands in the coastal zone

    The Noble-Abel Stiffened-Gas equation of state

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    International audienceHyperbolic two-phase flow models have shown excellent ability for the resolution of a wide range of applications ranging from interfacial flows to fluid mixtures with several velocities. These models account for waves propagation (acoustic and convective) and consist in hy-perbolic systems of partial differential equations. In this context, each phase is compressible and needs an appropriate convex equation of state (EOS). The EOS must be simple enough for intensive computations as well as boundary conditions treatment. It must also be accurate , this being challenging with respect to simplicity. In the present approach, each fluid is governed by a novel EOS named 'Noble Abel Stiffened Gas' (NASG), this formulation being a significant improvement of the popular 'Stiffened Gas' (SG) EOS. It is a combination of the so-called 'Noble-Abel' and 'Stiffened Gas' equations of state that adds repulsive effects to the SG formulation. The determination of the various thermodynamic functions and associated coefficients is the aim of this article. We first use thermodynamic considerations to determine the different state functions such as the specific internal energy, enthalpy and entropy. Then we propose to determine the associated coefficients for a liquid in the presence of its vapor. The EOS parameters are determined from experimental saturation curves. Some examples of liquid-vapor fluids are examined and associated parameters are computed with the help of the present method. Comparisons between analytical and experimental saturation curves show very good agreement for wide ranges of temperature for both liquid and vapor

    Steady one-dimensional nozzle flow solutions of liquid-gas mixtures

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    International audienceExact compressible one-dimensional nozzle flow solutions at steady state are determined in various limit situations of two-phase liquid-gas mixtures. First, the exact solution for a pure liquid nozzle flow is determined in the context of fluids governed by the compressible Euler equations and the `stiffened gas' equation of state. It is an extension of the well-known ideal-gas steady nozzle flow solution. Various two-phase flow models are then addressed, all corresponding to limit situations of partial equilibrium among the phases. The first limit situation corresponds to the two-phase flow model of Kapila et al. (Phys. Fluids, vol. 13, 2001, pp. 3002-3024), where both phases evolve in mechanical equilibrium only. This model contains two entropies, two temperatures and non-conventional shock relations. The second one corresponds to a two-phase model where the phases evolve in both mechanical and thermal equilibrium. The last one corresponds to a model describing a liquid-vapour mixture in thermodynamic equilibrium. They all correspond to two-phase mixtures where the various relaxation effects are either stiff or absent. In all instances, the various flow regimes (subsonic, subsonic-supersonic, and supersonic with shock) are unambiguously determined, as well as various nozzle solution profiles

    Oriented bilayers of a proteolipid complex, Annexin V – phospholipids, for solid state NMR analyses

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    We were able to obtain oriented bilayers in the presence of annexin V (35 kDa) and to measure structural and dynamic parameters. NMR results obtained so far show that annexin does not affect the structure of the membrane but modifies the dynamics of the phospholipids (T2e decrease). Our new method to prepare oriented biological samples, based on the preparation of small unilamellar proteolipid vesicles in biological buffers, should be suitable for any membrane protein – phospholipid complex and for solid state NMR or neutron diffraction experiments
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