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Estimates of DNA damage by the comet assay in the direct-developing frog Eleutherodactylus johnstonei (Anura, Eleutherodactylidae) Genet. Mol. Biol.
Valencia,Laura Carolina; García,Adriana; Ramírez-Pinilla,Martha Patricia; Fuentes,Jorge Luis.
The aim of this study was to use the Comet assay to assess genetic damage in the direct-developing frog Eleutherodactylus johnstonei. A DNA diffusion assay was used to evaluate the effectiveness of alkaline, enzymatic and alkaline/enzymatic treatments for lysing E. johnstonei blood cells and to determine the amount of DNA strand breakage associated with apoptosis and necrosis. Cell sensitivity to the mutagens bleomycin (BLM) and 4-nitroquinoline-1-oxide (4NQO) was also assessed using the Comet assay, as was the assay reproducibility. Alkaline treatment did not lyse the cytoplasmic and nuclear membranes of E. johnstonei blood cells, whereas enzymatic digestion with proteinase K (40 !g/mL) yielded naked nuclei. The contribution of apoptosis and necrosis...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bleomycin; Comet assay; DNA diffusion assay; Eleutherodactylus johnstonei; 4-nitroquinoline-1-oxide.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000400022
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Estimates of DNA strand breakage in bottlenose dolphin (Tursiops truncatus) leukocytes measured with the Comet and DNA diffusion assays Genet. Mol. Biol.
Díaz,Adriana; Carro,Sandra; Santiago,Livia; Estévez,Juan; Guevara,Celia; Blanco,Miriam; Sánchez,Laima; Sánchez,Liena; López,Nirka; Cruz,Danilo; López,Ronar; Cuetara,Elizabeth B.; Fuentes,Jorge Luis.
The analysis of DNA damage by mean of Comet or single cell gel electrophoresis (SCGE) assay has been commonly used to assess genotoxic impact in aquatic animals being able to detect exposure to low concentrations of contaminants in a wide range of species. The aims of this work were 1) to evaluate the usefulness of the Comet to detect DNA strand breakage in dolphin leukocytes, 2) to use the DNA diffusion assay to determine the amount of DNA strand breakage associated with apoptosis or necrosis, and 3) to determine the proportion of DNA strand breakage that was unrelated to apoptosis and necrosis. Significant intra-individual variation was observed in all of the estimates of DNA damage. DNA strand breakage was overestimated because a considerable amount...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Comet assay; DNA diffusion assay; DNA strand breakage; Tursiops truncatus.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572009000200025
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