Vanadium Toxicity Monitored by Fertilization Outcomes and Metal Related Proteolytic Activities in Paracentrotus lividus Embryos

Metal pharmaceutical residues often represent emerging toxic pollutants of the aquatic environment, as wastewater treatment plants do not sufficiently remove these compounds. Recently, vanadium (V) derivatives have been considered as potential therapeutic factors in several diseases, however, only limited information is available about their impact on aquatic environments. This study used sea urchin embryos (Paracentrotus lividus) to test V toxicity, as it is known they are sensitive to V doses from environmentally relevant to very cytotoxic levels (50 nM; 100 nM; 500 nM; 1 uM; 50 uM; 100 uM; 500 uM; and 1 mM).We used two approaches: The fertilization test (FT) and a protease detection assay after 36 h of exposure. V affected the fertilization percentage and increased morphological abnormalities of both egg and fertilization envelope, in a dose-dependent manner. Moreover, a total of nine gelatinases (with apparent molecular masses ranging from 309 to 22 kDa) were detected, and their proteolytic activity depended on the V concentration. Biochemical characterization shows that some of them could be aspartate proteases, whereas substrate specificity and the Ca2+/Zn2+ requirement suggest that others are similar to mammalian matrix metalloproteinases (MMPs)

Cellular Mechanotransduction: From Tension to Function

Living cells are constantly exposed to mechanical stimuli arising from the surrounding extracellular matrix (ECM) or from neighboring cells. The intracellular molecular processes through which such physical cues are transformed into a biological response are collectively dubbed as mechanotransduction and are of fundamental importance to help the cell timely adapt to the continuous dynamic modifications of the microenvironment. Local changes in ECM composition and mechanics are driven by a feed forward interplay between the cell and the matrix itself, with the first depositing ECM proteins that in turn will impact on the surrounding cells. As such, these changes occur regularly during tissue development and are a hallmark of the pathologies of aging. Only lately, though, the importance of mechanical cues in controlling cell function (e.g., proliferation, differentiation, migration) has been acknowledged. Here we provide a critical review of the recent insights into the molecular basis of cellular mechanotransduction, by analyzing how mechanical stimuli get transformed into a given biological response through the activation of a peculiar genetic program. Specifically, by recapitulating the processes involved in the interpretation of ECM remodeling by Focal Adhesions at cell-matrix interphase, we revise the role of cytoskeleton tension as the second messenger of the mechanotransduction process and the action of mechano-responsive shuttling proteins converging on stage and cell-specific transcription factors. Finally, we give few paradigmatic examples highlighting the emerging role of malfunctions in cell mechanosensing apparatus in the onset and progression of pathologies

DEFORMATION RESPONSE OF POLYDIMETHYLSILOXANE SUBSTRATES SUBJECTED TO UNIAXIAL QUASI-STATIC LOADING

To investigate cellular response of cardiomyocytes to substrate mechanics, biocompatible material with stiffness in physiological range is needed. PDMS based material is used for construction of microfluidic organ on chip devices for cell culture due to ease of device preparation, bonding, and possibility of surface functionalization. However it has stiffness orders of magnitude out of physiological range. Therefore, we adapted recently available protocol aiming to prepare substrates which offer stiffness in physiological range 5−100kPa using various mixtures of Sylgard. An in-house developed loading device with single micron position tracking accuracy and sub-micron position sensitivity was adapted for this experimental campaign. All batches of the samples were subjected to uniaxial loading. During quasi-static experiment the samples were compressed to minimally 40% deformation. The results are represented in the form of stress-strain curves calculated from the acquired force and displacement data and elastic moduli are estimated

Variabilidad genotípica y ambiental del peso, tamaño y aceite del grano de girasol (Helianthus annuus L.) en el Noreste argentino

La relación tegumento/embrión es uno de los principales factores que afectan el porcentaje de aceite en girasol. Mayores proporciones de embrión incrementa la cantidad de aceite del grano. En la actualidad, no es claro el efecto ambiental y la variabilidad genotípica en híbridos modernos de girasol de la relación tegumento/embrión y el porcentaje de aceite del grano. Los objetivos de este trabajo fueron i) determinar la variabilidad genotípica y ambiental para porcentaje de aceite y proporción de tegumento y embrión en el Noreste Argentino y ii) analizar la influencia de variables ambientales sobre la relación de tegumento/embrión y el porcentaje de aceite del grano de girasol. Se realizaron 4 experimentos durante la campaña 2014/2015 en las localidades de Reconquista (en dos fechas de siembra), Ceres y Villa Ocampo. Los tratamientos consistieron en una combinación de cuatro ambientes y quince híbridos comerciales de girasol. Las determinaciones realizadas fueron tipo de grano, tamaño (largo y ancho), peso de grano, peso y espesor del tegumento, peso del embrión, contenido de aceite de embrión y grano. Se realizaron análisis de componentes principales para explicar las relaciones de cada componente con las variables medidas. El porcentaje de aceite varió entre 55,2 y 43,3, mientras que el porcentaje de aceite de embrión varió entre 53,5 y 66,4%. En el porcentaje de aceite en el grano y relación tegumento/embrión se observaron diferencias significativas entre híbridos y ambientes. El rango obtenido en peso de grano fue de 29 a 91 mg. Uno de los factores ambientales más importantes que se relacionó con las variables fue la temperatura máxima y media durante el llenado de granos. Estos resultados contribuyen a la toma de decisiones sobre estabilidad y adaptación genotípica como así también herramienta para el mejoramiento genético para incrementar el porcentaje de aceite de futuros híbridos.The coat/embryo ratio is one of the main factors affecting the percentage of sunflower oil. Higher proportions of seed embryo increase the amount of oil in grain. Currently, environmental effect and genetic variability in modern hybrid sunflower coat/embryo ratio and the percentage of grain oil is unclear. The objectives of this study were i) determine the genotypic and environmental variability for oil percentage and ratio of seed coat and embryo in the northeastern regions of Argentina and ii) analyze the influence of environmental variables on the coat/embryo ratio and oil content sunflower grain. Four experiments during the 2014/2015 growing season were made in Reconquista (in two planting dates), Ceres and Villa Ocampo. The treatments consist in a combination of four environmental conditions by fifteen sunflower commercial hybrids. The determinations were grain type, size (length and width), whole grain weight, weight and thickness of the seed coat, embryo weight and oil content of embryo. Principal component analyses were performed to explain the relationships of each component with the measured variables. The percentage of oil ranged from 55,2 and 43,3%, while the percentage of seed embryo varied from 53,5 to 66,4%. The grain oil percentage and coat/embryo ratio showed significant differences among hybrids and ambient. The range obtained for grain weight ranged from 29 to 91 mg. One of the most important environmental factors related to the variables was maximum and mean temperature during grain filling. These results contribute to decide on adaptation and genotypic stability as well as a tool for breeding programs to increase the f oil percentage of future hybrids.EEA RafaelaFil: Rocca, Paola. Universidad Nacional de Villa María; ArgentinaFil: Martino, Laureano. Universidad Nacional de Villa María; ArgentinaFil: Rausch, Analia Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Rafaela. Agencia De Extensión Rural Ceres; ArgentinaFil: Zuil, Sebastian. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Reconquista; Argentin

A INTEGRAÇÃO ENSINO-SERVIÇO NA REDE DE ATENÇÃO PRIMÁRIA DA GERÊNCIA GLÓRIA/CRUZEIRO/CRISTAL DE PORTO ALEGRE/RS: UNIDADE DE SAÚDE GLÓRIA

Este número do Boletim Informativo Saberes Plurais apresenta informações sobre A Integração Ensino-Serviço na Rede de Atenção Primária da Gerência Glória/Cruzeiro/Cristal: Unidade de Saúde Glória no município de Porto Alegre/RS, que se originaram do projeto de pesquisa “Avaliação de Redes Integradas de Atenção e Ensino na Saúde do Sistema Único de Saúde”, financiado pelo Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) - Edital da Chamada Universal MCTI/CNPq 01/2016 (termo de concessão 42430/2016-3). Por meio de um material de fácil manuseio e compreensão, pretende-se possibilitar o reconhecimento das relações entre processos de Integração Ensino-Serviço e a constituição das redes de Atenção Primária à Saúde. A pesquisa se constituiu em um estudo de caso com abordagem qualitativa. Participaram 16 profissionais de saúde: 1 médico, 2 enfermeiros, 1 cirurgião-dentista, 6 técnicos de enfermagem, 1 auxiliar de saúde bucal, 5 agentes comunitários de saúde; e 2 estudantes da graduação. A condução dos debates foi apoiada por um roteiro de pesquisa fundamentado nas dimensões das redes de saúde. Conclui-se que processos de Integração Ensino-Serviço produzem Educação Permanente em Saúde demonstrando-se como apoio nos processos de territorialização e consolidação das redes, entretanto limitações ainda são identificadas nos sistemas de governança para a integração das redes de atenção e ensino

Top-down structures of mafic enclaves within the Valle Fértil magmatic complex (Early Ordovician, San Juan, Argentina)

Magmatic structures related to the mechanical interaction between mafic magmas and granitoids have been studied in the Valle Fértil calc-alkaline igneous complex, Argentina. Excepcional outcrops with vertical walls of more than 300 m high allow us the study of three-dimensional geometries of individual blobs of mafic magma as well as the geometry of pipe-like structures in which mafic microgranular enclaves are concentrated in more than 50 times the normal abundance in the granodiorite mass. The shape of enclaves and pipe-like structures are interpreted as the ressult of top-to-down intrusions of a mafic magma into a granodiorite-tonalite mass. These sinking structures are the result of a reverselly stratified magma chamber with gabbros and diorites at the top and granodiorite-tonalite at the bottom. They may account for most of the structures found in microgranular enclaves and magma mingling zones that characterize calc-alkaline batholiths. Synplutonic intrusions from the top is the only plausible mechanism to account for the observed structures. The model may be of general application to calc-alkaline batholiths characterized by the presence of mafic microgranular enclaves. An implication of these reverselly stratified magma chambers is the presence of a petrological inversion which may be the consequence of cold diapirs emplaced below the mantle wedge in a suprasubduction setting

Dissecting the intracellular signalling and fate of a DNA nanosensor by super-resolution and quantitative microscopy

DNA nanodevices have been developed as platforms for the manipulation of gene expression, delivery of molecular payloads, and detection of various molecular targets within cells and in other complex biological settings. Despite efforts to translate DNA nanodevices from the test tube (in vitro) to living cells, their intracellular trafficking and functionality remain poorly understood. Herein, quantitative and super-resolution microscopy approaches were employed to track and visualise, with nanometric resolution, the molecular interactions between a synthetic DNA nanosensor and transcription factors in intracellular compartments. Specifically, fluorescence resonance energy transfer microscopy, fluorescence correlation spectroscopy, fluorescence lifetime imaging microscopy and multicolour single-molecule localisation microscopy were employed to probe the specific binding of the DNA nanosensor to the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-kappa B). We monitored the mobility, subcellular localisation and degradation of the DNA nanosensor inside living prostate cancer PC3 cells. Super-resolution imaging enabled the direct visualisation of the molecular interactions between the synthetic DNA nanosensors and the NF-kappa B molecules in cells. This study represents a significant advance in the effective detection as well as understanding of the intracellular dynamics of DNA nanosensors in a complex biological milieu

Clinical and Pharmacological Aspects of Inflammatory Demyelinating Diseases in Childhood: An Update

Inflammatory demyelinating diseases comprise a spectrum of disorders affecting the myelin of the central and peripheral nervous system. These diseases can usually be differentiated on the basis of clinical, radiological, laboratory and pathological findings

Novel bicistronic lentiviral vectors correct β-Hexosaminidase deficiency in neural and hematopoietic stem cells and progeny: implications for in vivo and ex vivo gene therapy of GM2 gangliosidosis.

Abstract The favorable outcome of in vivo and ex vivo gene therapy approaches in several Lysosomal Storage Diseases suggests that these treatment strategies might equally benefit GM2 gangliosidosis. Tay-Sachs and Sandhoff disease (the main forms of GM2 gangliosidosis) result from mutations in either the HEXA or HEXB genes encoding, respectively, the α- or β-subunits of the lysosomal β-Hexosaminidase enzyme. In physiological conditions, α- and β-subunits combine to generate β-Hexosaminidase A (HexA, αβ) and β-Hexosaminidase B (HexB, ββ). A major impairment to establishing in vivo or ex vivo gene therapy for GM2 gangliosidosis is the need to synthesize the α- and β-subunits at high levels and with the correct stoichiometric ratio, and to safely deliver the therapeutic products to all affected tissues/organs. Here, we report the generation and in vitro validation of novel bicistronic lentiviral vectors (LVs) encoding for both the murine and human codon optimized Hexa and Hexb genes. We show that these LVs drive the safe and coordinate expression of the α- and β-subunits, leading to supranormal levels of β-Hexosaminidase activity with prevalent formation of a functional HexA in SD murine neurons and glia, murine bone marrow-derived hematopoietic stem/progenitor cells (HSPCs), and human SD fibroblasts. The restoration/overexpression of β-Hexosaminidase leads to the reduction of intracellular GM2 ganglioside storage in transduced and in cross-corrected SD murine neural progeny, indicating that the transgenic enzyme is secreted and functional. Importantly, bicistronic LVs safely and efficiently transduce human neurons/glia and CD34+ HSPCs, which are target and effector cells, respectively, in prospective in vivo and ex vivo GT approaches. We anticipate that these bicistronic LVs may overcome the current requirement of two vectors co-delivering the α- or β-subunits genes. Careful assessment of the safety and therapeutic potential of these bicistronic LVs in the SD murine model will pave the way to the clinical development of LV-based gene therapy for GM2 gangliosidosis