285 research outputs found

    Functional Analysis of Vaccinia Virus B5R Protein: Role of the Cytoplasmic Tail

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    AbstractVaccinia extracellular enveloped virus (EEV) is important for cell-to-cell and long-range virus spread bothin vitroandin vivo. Six genes have been identified that encode protein constituents of the EEV outer membrane, and some of these proteins are critical for EEV formation. The B5R gene encodes an EEV-specific type I membrane protein, and deletion of this gene markedly decreases EEV formation and results in a small plaque phenotype. Data suggest that the transmembrane domain, cytoplasmic tail, or both contain the EEV localization signals that are required for targeting of the B5R protein to EEV and for EEV formation. Here, we report the construction of mutant vaccinia viruses in which the wild-type B5R gene was replaced with a mutated one that encodes a protein with the putative cytoplasmic tail deleted. The mutated protein showed normal intracellular distribution and was properly incorporated into EEV. Vaccinia viruses expressing the B5R protein lacking the cytoplasmic tail formed plaques that were similar in type and size to those formed by wild-type viruses and produced equivalent amounts of infectious EEV. These results indicate that the B5R cytoplasmic tail is not necessary for EEV formation and points to the transmembrane domain as the major determinant for targeting the B5R protein to the outer membrane of EEV and for supporting EEV formation

    Extraction of sulfated polysaccharides by autohydrolysis of brown seaweed Fucus vesiculosus

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    The extraction of sulfated polysaccharides (fucoidan) by autohydrolysis (AH) of brown seaweed Fucus vesiculosus was studied. Experimental assays were performed under different conditions of temperature (160 to 200°C) and reaction time (10 to 30 min) according to a 22 central composite design, and the conditions able to maximize the fucoidan yield were selected. The alga degradation and the total sugar yield in the liquor after AH were also determined to each experimental condition. The highest fucoidan yield (∼16.5% w/w) was obtained when the AH process was performed at 180°C for 20 min. This product was characterized by high-performance liquid chromatography, infrared analysis spectroscopy, and thermal gravimetric analyses, which verified the presence of fucose and galactose as main components (70:30% mol ratio, in average) and an SO3 content higher than 20%. AH process under optimum reaction conditions was an effective method to recover fucoidan from F. vesiculosus. The use of this technology brings also important advantages from economical and environmental viewpoints since it does not require the use of chemical solvent and generates less waste when compared to conventional extraction procedures.R.M. Rodriguez-Jasso would like to thank the Mexican Science and Technology Council (CONACYT, Mexico) for Ph.D. fellowship support (grant number 213592/308679). The authors also thank Professor Isabel Sousa Pinto, Rita Araujo PhD from CII-MAR, University of Porto, and Bartolomeu de Souza PhD for the collaboration during the selection and collection of the seaweed used in this study

    Microwave-assisted extraction of sulfated fucans from brown seaweed and evaluation of fungal strains for enzymesactive production toward this class of polysaccharide

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    Sulphated polysaccharides from brown seaweedss comprise a complex group of macromolecules with a wide range of important biological properties such as anticoagulant, antioxidant, antiproliferative, antitumoral, anticomplementary, anti-inflammatory, antiviral, antipeptic and antiadhesive activities. Fucoidan is one of the main sulphated fucan, mostly interesting for their biological activities specially the potential to inhibit HIV reverse transcriptase and the possible application as active compound in antiretroviral drugs. However, algae remain largely unexploited and seaweeds can be found in sufficient amount for the commercial exploitation. Usually, most of the processes to recover sulfated polysaccharides from natural sources consist in acid extractions during long reaction times. Specific enzymes able to degrade fucoidan matrix (fucoidanases) are important tools to establish structural characteristics and biological functions of this polysaccharide. Such enzymes, have been only isolated from marine organisms. Reports of fungal microorganisms with enzymatic activity over this sulfated-polysaccharide are scarce. The aims of the present work were: 1) to recovery of sulphated polysaccharides (fucoidan) by microwave-assisted extraction under different operational conditions and 2) the identification of fungal strains able to growth over fucoidan-based media and to produce active fucoidanases

    Cosmology of a brane radiating gravitons into the extra dimension

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    We study in a self-consistent way the impact of the emission of bulk gravitons on the (homogeneous) cosmology of a three-brane embedded in a five-dimensional spacetime. In the low energy regime, we recover the well known result that the bulk affects the Friedmann equation only via a radiation-like term \C/a^4, called dark or Weyl radiation. By contrast, in the high energy regime, we find that the Weyl parameter \C is no longer constant but instead grows very rapidly as \C\propto a^4. As a consequence, the value of \C today is not a free parameter as usually considered but is a fixed number, which, generically, depends only on the number of relativistic degrees of freedom at the high/low energy transition. Our estimated amount of Weyl radiation satisfies the present nucleosynthesis bounds.Comment: 12 page

    Effects of feed restriction during pregnancy on maternal reproductive outcome, foetal hepatic IGF gene expression and offspring performance in the rabbit

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    Primiparous female rabbits have high nutritional requirements and, while it is recommended that they are subjected to an extensive reproductive rhythm, this could lead to overweight, affecting reproductive outcomes. We hypothesised that restricting food intake during the less energetic period of gestation could improve reproductive outcome without impairing offspring viability. This study compares two groups of primiparous rabbit does in an extensive reproductive programme, one in which feed was restricted from Day 0 to Day 21 of gestation (R021), and another in which does were fed ad libitum (control) throughout pregnancy. The mother and offspring variables compared were (1) mother reproductive outcomes at the time points pre-implantation (Day 3 postartificial insemination [AI]), preterm (Day 28 post-AI) and birth; and (2) the prenatal offspring characteristic IGF system gene expression in foetal liver, liver fibrosis and foetus sex ratio, and postnatal factor viability and growth at birth, and survival and growth until weaning. Feed restriction did not affect the conception rate, embryo survival, or the number of morulae and blastocysts recovered at Day 3 post-AI. Preterm placenta size and efficiency were similar in the two groups. However, both implantation rate (P < 0.001) and the number of foetuses (P = 0.05) were higher in the R021 mothers than controls, while there was no difference in foetal viability. Foetal size and weight, the weights of most organs, organ weight/BW ratios and sex ratio were unaffected by feed restriction; these variables were only affected by uterine position (P < 0.05). Conversely, in the R021 does, foetal liver IGBP1 and IGF2 gene expression were dysregulated despite no liver fibrosis and a normal liver structure. No effects of restricted feed intake were produced on maternal fertility, prolificacy, or offspring birth weight, but control females weaned more kits. Litter weight and mortality rate during the lactation period were also unaffected. In conclusion, pre-implantation events and foetal development were unaffected by feed restriction. While some genes of the foetal hepatic IGF system were dysregulated during pregnancy, liver morphology appeared normal, and the growth of foetuses and kits until weaning was unmodified. This strategy of feed restriction in extensive reproductive rhythms seems to have no significant adverse effects on dam reproductive outcome or offspring growth and viability until weaning

    Sediment characterization of an artificial reservoir impacted by mining activity in the Almadén mercury district (Spain)

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    Los sedimentos del embalse Castilseras, ubicado aguas abajo en el río Valdeazogues en el distrito de Almadén, se recogieron para evaluar los contenidos en mercurio totales asociados con las aportaciones de sedimentos del río que provienen de las minas clausuradas. Se ha determinado el contenido en mercurio total así como otras propiedades físico-químicas de estos sedimentos. Los resultados muestran que el contenido en mercurio es elevado, y que se relaciona con el de los materiales de la zona, relacionados con la minería. Los sedimentos presentan un nivel de acidez significativo, son moderadamente salinos y tienen un tamaño de grano predominantemente limoso. El análisis estadístico indica que las concentraciones de mercurio están positivamente correlacionadas con el porcentaje en limo de las muestras. De los resultados obtenidos se puede concluir que el contenido en mercurio se va diluyendo aguas abajo del embalse y que éste está relacionado con la actividad minera, convirtiendo a Castilseras en uno de los embalses con mayor contenido en mercurio del mundo.Sediments from the Castilseras reservoir, located downstream of the Valdeazogues River in the Almadén mercury district were collected to assess the total mercury content associated with river sediment input from several decommissioned mines. A geochemical characterization, including the determination of total mercury content, were investigated using different physical and chemical techniques. The results obtained suggested that sediments are slightly acidic and characterized by high mercury, related to mining activities. In addition, sediments showed a silty texture and the electrical conductivity results showed moderate salinity, related to acid mine drainage (AMD) processes. The multivariate statistical analysis suggested that mercury content is positively correlated with the silt fraction in collected samples. In regard to the obtained results, we can conclude that Castilseras is one of the reservoirs with the highest mercury concentration, as a result of mining activities carried out in the zone. In this reservoir, the mercury content is diluted downstreamDepto. de Mineralogía y PetrologíaFac. de Ciencias GeológicasTRUEpu
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