4 resultados para target sites

em Repositório Institucional UNESP - Universidade Estadual Paulista "Julio de Mesquita Filho"


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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Introduced exotic species cause environmental changes and threat public health in target sites. Illegal trade has enhanced this problem. To first report these risks in Brazil, exotic snakes found in São Paulo City (SPC) (23degrees32' S, 46degrees38' W), southeastern Brazil, and sent to Instituto Butantan between 1995 and 2000, were listed and characterized by their biological attributes. Seventy-six individuals of sixteen alien species were collected. Euriecians snakes, mainly booids, were predominant. Using multivariate techniques, their ecological niches were compared to those of 26 native species, as a way to point out the resource's availability. To evaluate the potential of successful implantation, two species absent in SPC and considered as problem snakes are included in these analyses: the brown treesnake Boiga irregularis and the habu Trimeresurus flavoviridis. There were niche similarities between these pest snakes, exotic booids and native viperids largely due to the similarities in the chosen prey (mammals), diel activity (nocturnal), color pattern (variegated) and body size (medium to large). To avoid predictable undesirable effects of implanted pest snakes, traffic control and punishment should be improved, as well as parallel environmental education programs.

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We report a method for studying global DNA methylation based on using bisulfite treatment of DNA and simultaneous PCR of multiple DNA repetitive elements, such as Alu elements and long interspersed nucleotide elements (LINE). The PCR product, which represents a pool of approximately 15000 genomic loci, could be used for direct sequencing, selective restriction digestion or pyrosequencing, in order to quantitate DNA methylation. By restriction digestion or pyrosequencing, the assay was reproducible with a standard deviation of only 2% between assays. Using this method we found that almost two-thirds of the CpG methylation sites in Alu elements are mutated, but of the remaining methylation target sites, 87% were methylated. Due to the heavy methylation of repetitive elements, this assay was especially useful in detecting decreases in DNA methylation, and this assay was validated by examining cell lines treated with the methylation inhibitor 5-aza-2'deoxycytidine (DAC), where we found a 1-16% decrease in Alu element and 18-60% LINE methylation within 3 days of treatment. This method can be used as a surrogate marker of genome-wide methylation changes. In addition, it is less labor intensive and requires less DNA than previous methods of assessing global DNA methylation.

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P>The reactivity of sera collected from guinea pigs after three infestations with Amblyomma triste nymphs on histological sections of the same tick species was investigated through immunohistochemistry to identify potential target cells and tissues. Six guinea pigs were infested thrice, at 30 day intervals, with 30 nymphs of A. triste per animal per infestation. Blood samples were collected from the guinea pigs 15 days after each infestation for serum separation; normal serum was obtained before the first infestation as control. Unfed A. triste nymphs' histological sections were submitted to indirect immunohistochemistry technique by using normal or hyperimmune guinea pig serum as primary antibody and a goat IgG-alkaline phosphatase-APase conjugate as secondary antibody. A weak to moderate APase activity was observed in cells of salivary glands, midgut and haemolymph of unfed nymphs incubated with hyperimmune serum.