2,003 research outputs found

    The yeast prion Ure2p native-like assemblies are toxic to mammalian cells regardless of their aggregation state.

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    The yeast prion Ure2p assembles in vitro into oligomers and fibrils retaining the alpha-helix content and binding properties of the soluble protein. Here we show that the different forms of Ure2p native-like assemblies (dimers, oligomers, and fibrils) are similarly toxic to murine H-END cells when added to the culture medium. Interestingly, the amyloid fibrils obtained by heat treatment of the toxic native-like fibrils appear harmless. Moreover, the Ure2p C-terminal domain, lacking the N-terminal segment necessary for aggregation but containing the glutathione binding site, is not cytotoxic. This finding strongly supports the idea that Ure2p toxicity depends on the structural properties of the flexible N-terminal prion domain and can therefore be considered as an inherent feature of the protein, unrelated to its aggregation state but rather associated with a basic toxic fold shared by all of the Ure2p native-like assemblies. Indeed, the latter are able to interact with the cell surface, leading to alteration of calcium homeostasis, membrane permeabilization, and oxidative stress, whereas the heat-treated amyloid fibrils do not. Our results support the idea of a general mechanism of toxicity of any protein/peptide aggregate endowed with structural features, making it able to interact with cell membranes and to destabilize them. This evidence extends the widely accepted view that the toxicity by protein aggregates is restricted to amyloid prefibrillar aggregates and provides new insights into the mechanism by which native-like oligomers compromise cell viability

    An operational model for capturing grape ripening dynamics to support harvest decisions

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    Grape ripening is a critical phenological phase during which many metabolites that impact wine quality accumulate in the berries. Major changes in berry composition include a rapid increase in sugar and a decrease in malic acid content and concentration. Its duration is highly variable depending on grapevine variety, climatic parameters, soil type and management practices. Together with the timing of mid-veraison, this duration determines when grapes can be harvested. Viticulturists and winemakers monitor the sugar-to-total acidity ratio (S/TA) during grape ripening and start harvesting grapes when this ratio reaches the optimum value for the desired wine style. The S/TA ratio evolves linearly as a function of thermal summation during the first four weeks following the onset of ripening. The linearity of the evolution of the S/TA ratio as a function of thermal time during the first four weeks following mid-veraison is applied in this study on two large data sets encompassing (1) 53 varieties studied during 10 years with two to four replicates for each combination of year and cultivar and (2) two varieties, cultivated on three soil types over 13 years. Grape ripening speed is highly variable. The effects of the year impact ripening speed more than the effects of the soil or the variety, although all three effects are highly significant. Grape ripening speed decreases with berry weight and also varies with vine water status. By using this approach, viticulturists and winemakers can assess four weeks after mid-veraison, for each individual vineyard parcel, at what speed grape ripening progresses. Combined with precise mid-veraison scoring, expertise from previous vintages and complementary approaches like sensory assessment of berries, it allows harvest date estimates to be fine-tuned. The results of this study can also be used to identify slow ripening varies, which are better performing in warm climates and, thus, better adapted to climate change

    The Development of a European and Mediterranean Chickpea Association Panel (EMCAP)

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    Association panels represent a useful tool for quantitative trait loci (QTL) mapping and pre-breeding. In this study, we report on the development of a European and Mediterranean chickpea association panel as a useful tool for gene discovery and breeding. Chickpea (Cicer arietinum L.) is one of the most important food legumes worldwide and a key crop in the Mediterranean environments. The selection of genotypes followed criteria aimed to build a set of materials representative of the genetic diversity of chickpea germplasm focusing on the European and Mediterranean environments, which have largely been ignored to date. This tool can help breeders to develop novel varieties adapted to European and Mediterranean agro-ecosystems. Initially, 1931 chickpea accessions were phenotypically evaluated in a field trial in central Italy. From these, an association panel composed by 480 genotypes derived from single-seed descent was identified and phenotypically evaluated. Current and future phenotypic data combined with the genotypic characterization of the association panel will allow to dissect the genetic architecture of important adaptive and quality traits and accelerate breeding. This information can be used to predict phenotypes of unexploited chickpea genetic resources available in genebanks for breeding

    Dendritic cells infiltrate the cardiac muscle fibers during myocardial infarction

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    Myocardial infarction (MI) consists in myocardial cell death due to prolonged ischemia. Partial ischemia at the periphery at the necrotic area may lead to “hibernating” myocardium, which may eventually recover. Upon necrosis an inflammatory a process starts [1], leading to healing through formation of a fibrous scar. Dendritic cells (DC) are involved in the regulation of immune responses and in the organization of inflammatory cell infiltrates in vascular wall, even independent of immune reactions. Another cell type involved in acute reaction to tissue injury are mast cells. The behaviour of DC and mast cells in myocardial infarction is still to be studied. To address this issue myocardial samples were taken at autopsy from the left ventricle of subjects respectively affected by (1) coronarosclerosis, (2) acute MI, (3) previous MI, and (4) traumatic lesions assumed as controls. Cryosections were stained with haematoxilin heosin and by immunohistochemistry. Fiber alterations consisting in loss of acidophilia and disappearance of nuclei and intercalar disks were found only in acute MI, while a cell infiltrate was found both in acute and previous MI. Massive infiltration of DC was found only in acute MI, while mast cells were similar to controls. These preliminary results suggest that DC react early to myocardial injury and therefore may be candidate regulators of the inflammatory and scarring response in this tissue and markers of acute myocardial infarction
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