263 research outputs found

    The molecular chaperone Hsp90 is a component of the cap-binding complex and interacts with the translational repressor Cup during Drosophila oogenesis

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    In metazoa, the spatio-temporal translation of diverse mRNAs is essential to guarantee proper oocyte maturation and early embryogenesis. The eukaryotic translation initiation factor 4E (eIF4E), which binds the 5′ cap structure of eukaryotic mRNAs, associates with either stimulatory or inhibitory factors to modulate protein synthesis. In order to identify novel factors that might act at the translational level during Drosophila oogenesis, we have undertaken a functional proteomic approach and isolated the product of the Hsp83 gene, the evolutionarily conserved chaperone Hsp90, as a specific component of the cap-binding complex. Here we report that Hsp90 interacts in vitro with the translational repressor Cup. In addition, we show that Hsp83 and cup interact genetically, since lowering Hsp90 activity enhances the oogenesis alterations linked to diverse cup mutant alleles. Hsp90 and Cup co-localize in the cytoplasm of the developing germ-line cells within the germarium, thus suggesting a common function from the earliest stages of oogenesis. Taken together, our data start elucidating the role of Hsp90 during Drosophila female germ-line development and strengthen the idea that Cup has multiple essential functions during egg chamber development

    Fabrication of CZTSe/CIGS Nanowire Arrays by One-Step Electrodeposition for Solar-Cell Application

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    The paper reports some preliminary results concerning the manufacturing process of CuZnSnSe (CZTSe) and CuInGaSe (CIGS) nanowire arrays obtained by one-step electrodeposition for p-n junction fabrication. CZTSe nanowires were obtained through electrodeposition in a polycarbonate membrane by applying a rectangular pulsed current, while their morphology was optimized by appropriately setting the potential and the electrolyte composition. The electrochemical parameters, including pH and composition of the solution, were optimized to obtain a mechanically stable array of nanowires. The samples were characterized by scanning electron microscopy, Raman spectroscopy, and energy-dispersion spectroscopy. The nanostructures obtained showed a cylindrical shape with an average diameter of about 230 nm and a length of about 3 m, and were interconnected due to the morphology of the polycarbonate membrane. To create the p-n junctions, first a thin film of CZTSe was electrodeposited to avoid direct contact between the ZnS and Mo. Subsequently, an annealing process was carried out at 500 °C in a S atmosphere for 40 min. The ZnS was obtained by chemical bath deposition at 95 °C for 90 min. Finally, to complete the cell, ZnO and ZnO:Al layers were deposited by magnetron-sputtering

    A structure-based approach for novel immunodiagnostics targeting Trypanosoma cruzi and Schistosoma spp.

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    The tropical diseases Chagas Disease and Schistosomiasis, caused by the parasites Trypanosoma cruzi and Schistosoma spp, respectively, are often misdiagnosed and if untreated can be fatal. Indeed, their diagnosis presents a current challenge, due to difficulties in recognizing clinical symptoms and the lack of specific and reliable diagnostic tools. Highest prevalence is respectively in Latin America and Central Africa. However, massive human migration from endemic areas and travelling to tropical regions, contribute to the diffusion of these diseases in non-endemic areas, increasing the need of rapid diagnostic tools and prevention strategies. The aim of this project is to develop rapid, peptide-based microarrays that present multiple immunoreactive epitopes predicted from 3D protein structures of antigens from several pathogens, to be applied in point-of-care diagnostic kits for use in the Lombardy area of Italy. This approach showed promising results when applied to antigens from bacterial pathogens, in particular Burkholderia pseudomallei that led to the identification of epitopes with both diagnostic and therapeutic potential. Putative antigens selected from both parasites will be overexpressed, purified and crystallized for 3D structure studies. Based on the determined 3D protein structures, Molecular Dynamics (MD) analyses will be carried out and epitope predictions will be made on output MD structures using the Matrix of Local Coupling Energies method that detects residues located in conformationally dynamic regions of the protein surface. Predicted reactive epitopes will be synthetized as peptides, tested for immunoreactivity against immune sera from infected patients and used for antibody production. Selected immunoreactive synthetic peptides will be immobilised in specific orientations, using click chemistry, on a polymeric coated microarray chip. Sensitivity and specificity of the chip will be evaluated with sera of infected patients

    Two Novel Fish Paralogs Provide Insights Into the Rid Family of Imine Deaminases Active in Pre-Empting enamine/imine Metabolic Damage

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    Reactive Intermediate Deaminase (Rid) protein superfamily includes eight families among which the RidA is conserved in all domains of life. RidA proteins accelerate the deamination of the reactive 2-aminoacrylate (2AA), an enamine produced by some pyridoxal phosphate (PLP)-dependent enzymes. 2AA accumulation inhibits target enzymes with a detrimental impact on fitness. As a consequence of whole genome duplication, teleost fish have two ridA paralogs, while other extant vertebrates contain a single-copy gene. We investigated the biochemical properties of the products of two paralogs, identified in Salmo salar. SsRidA-1 and SsRidA-2 complemented the growth defect of a Salmonella enterica ridA mutant, an in vivo model of 2AA stress. In vitro, both proteins hydrolyzed 2-imino acids (IA) to keto-acids and ammonia. SsRidA-1 was active on IA derived from nonpolar amino acids and poorly active or inactive on IA derived from other amino acids tested. In contrast, SsRidA-2 had a generally low catalytic efficiency, but showed a relatively higher activity with IA derived from L-Glu and aromatic amino acids. The crystal structures of SsRidA-1 and SsRidA-2 provided hints of the remarkably different conformational stability and substrate specificity. Overall, SsRidA-1 is similar to the mammalian orthologs whereas SsRidA-2 displays unique properties likely generated by functional specialization of a duplicated ancestral gene

    The GINGER Project and status of the ring-laser of LNGS

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    A ring-laser attached to the Earth measures the absolute angular velocity of the Earth summed to the relativistic precessions, de Sitter and Lense-Thirring. GINGER (Gyroscopes IN GEneral Relativity) is a project aiming at measuring the LenseThirring effect with a ground based detector; it is based on an array of ring-lasers. Comparing the Earth angular velocity measured by IERS and the measurement done with the GINGER array, the Lense-Thirring effect can be evaluated. Compared to the existing space experiments, GINGER provides a local measurement, not the averaged value and it is unnecessary to model the gravitational field. It is a proposal, but it is not far from being a reality. In fact the GrossRing G of the Geodesy Observatory of Wettzell has a sensitivity very close to the necessary one. G ofWettzell is part of the IERS system which provides the measure of the Length Of the DAY (LOD); G provides information on the fast component of LOD. In the last few years, a roadmap toward GINGER has been outlined. The experiment G-GranSasso, financed by the INFN Commission II, is developing instrumentations and tests along the roadmap of GINGER. In this short paper the main activities of G-GranSasso and some results will be presented. The first results of GINGERino will be reported, GINGERino is the large ring-laser installed inside LNGS and now in the commissioning phase. Ring-lasers provide as well important informations for geophysics, in particular the rotational seismology, which is an emerging field of science. GINGERino is one of the three experiments of common interest between INFN and INGV

    Measurement results and improvements on an open EPR system

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    Electron spin resonance (ESR) is a spectroscopic method that allows to measure stable radicals induced by ionizing radiation. The EPR measurements can help to estimate the dose absorbed by people exposed during a nuclear disaster, detecting the number of radicals induced in their mobile phones due to the exposition [1]. Using conventional closed microwave cavities, the phone display must be fragmented in order to be introduced inside the resonator, becoming no more usable. The aim of this work is to develop a system, compatible with the spectrometer Bruker Elexys E500, able to preserve the sample integrity. The system uses an X-band resonant metallic cavity with a slit, realized on one side, for the leak of the excitation magnetic field and a Helmotz coil pair. The resonator allows measuring a sample lodged outside the cavity, while the coils produce a 100 kHz modulated field that encodes the output signal at a particular frequency and increases the SNR
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