946 resultados para lineage


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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Single nucleotide polymorphisms (SNPs) were investigated in eighteen genes of sixteen populations of Aedes aegypti in Brazil. Eight SNP markers were selected in nine genes and surveyed in A. aegypti populations of three localities in different geographical locations. SNPs revealed significant genetic differentiation among populations recently analyzed by mitochondria DNA (mtDNA) and represented by a single genetic group (lineage). Results suggest that a haplotype derived from mtDNA analysis could be represented by different Aedes lineages revealed by SNP characterization. Genetic distances (pairwise F(ST)), AMOVA and cluster analyses indicated a high genetic structure for the A. aegypti populations investigated by SNPs. This set of SNP markers represents a useful tool for genetic studies in A. aegypti populations

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Background: Biopharmaceutical drugs are mainly recombinant proteins produced by biotechnological tools. The patents of many biopharmaceuticals have expired, and biosimilars are thus currently being developed. Human granulocyte colony stimulating factor (hG-CSF) is a hematopoietic cytokine that acts on cells of the neutrophil lineage causing proliferation and differentiation of committed precursor cells and activation of mature neutrophils. Recombinant hG-CSF has been produced in genetically engineered Escherichia coli ( Filgrastim) and successfully used to treat cancer patients suffering from chemotherapy-induced neutropenia. Filgrastim is a 175 amino acid protein, containing an extra N-terminal methionine, which is needed for expression in E. coli. Here we describe a simple and low-cost process that is amenable to scaling-up for the production and purification of homogeneous and active recombinant hG-CSF expressed in E. coli cells.Results: Here we describe cloning of the human granulocyte colony-stimulating factor coding DNA sequence, protein expression in E. coli BL21(DE3) host cells in the absence of isopropyl-beta-D-thiogalactopyranoside ( IPTG) induction, efficient isolation and solubilization of inclusion bodies by a multi-step washing procedure, and a purification protocol using a single cationic exchange column. Characterization of homogeneous rhG-CSF by size exclusion and reverse phase chromatography showed similar yields to the standard. The immunoassay and N-terminal sequencing confirmed the identity of rhG-CSF. The biological activity assay, in vivo, showed an equivalent biological effect (109.4%) to the standard reference rhG-CSF. The homogeneous rhG-CSF protein yield was 3.2 mg of bioactive protein per liter of cell culture.Conclusion: The recombinant protein expression in the absence of IPTG induction is advantageous since cost is reduced, and the protein purification protocol using a single chromatographic step should reduce cost even further for large scale production. The physicochemical, immunological and biological analyses showed that this protocol can be useful to develop therapeutic bioproducts. In summary, the combination of different experimental strategies presented here allowed an efficient and cost-effective protocol for rhG-CSF production. These data may be of interest to biopharmaceutical companies interested in developing biosimilars and healthcare community.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The description of patterns of variation in any character system within well-defined species is fundamental for understanding lineage diversification and the identification of geographic units that represent opportunities for sustained evolutionary divergence. In this paper, we analyze intraspecific variation in cranial shape in the Pumpkin Toadlet, Brachycephalus ephippium-a miniaturized species composed of isolated populations on the slopes of the mountain ranges of southeastern Brazil. Shape variables were derived using geometric-statistical methods that describe shape change as localized deformations in a spatial framework defined by anatomical landmarks in the cranium of B. ephippium. By statistically weighting differences between landmarks that are not close together (changes at larger geometric scale), cranial variation among geographic samples of B. ephippium appears continuous with no obvious gaps. This pattern of variation is caused by a confounding effect between within-sample allometry and among-sample shape differences. In contrast, by statistically weighting differences between landmarks that are at close spacing (changes at smaller geometric scale), differences in shape within- and among-sample variation are not confounded, and a marked geographic differentiation among population samples of B. ephippium emerges. The observed pattern of geographic differentiation in cranial shape apparently cannot be explained as isolation-by-distance. This study provides the first evidence that the detection of morphological variation or lack thereof, that is, morphological conservatism, may be conditional on the scale of measurement of variation in shape within the methodological formalism of geometric morphometrics.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Morphometric data for 30 species of swarming wasps (Vespidae: Polistinae: Epiponini) are presented, representing all currently recognized genera. Data are coded according to whether females that were shown by dissection to be egglayers are larger, similar, or smaller for each dimension than non-egglayers. These data are analysed in a phylogenetic framework with primitively social Polistes and Mischocyttarus as outgroups. Representative measurements are illustrated to show that most genera of Epiponini appear to have ancestry in a lineage that has no queen caste comparable with either the primitively social outgroups, or the more derived species of the tribe. This analysis indicates that a conspiracy of workers that operates without a queen characterizes the societies of many Epiponini, or their recent ancestors. (c) 2008 The Linnean Society of London.