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Assessment of the oxidative metabolism of calves’ bronchoalveolar cells: comparison between NBT and fluorimetric techniques

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Detalhes bibliográficos
Resumo:ABSTRACT: Evaluation of alveolar macrophage functions of cattle is an important tool in order to assess whether measures taken during the cattle husbandry can decrease the respiratory tract defense. The aim of this study was to determine whether dexamethasone used at therapeutic dose can affect the oxidative metabolism of alveolar macrophages of cattle. This was evaluated by two tests, the fluorometric and colorimetric. The similarity of the results was studied, using alveolar macrophages of six healthy cattle, obtained from bronchoalveolar lavage on a basal and an immunosuppressant moment after the application of dexamethasone. For the fluorometric test, alveolar macrophages were incubated with Staphylococcus aureus and 2’-7’dichlorohidroflurescein, and analyzed by flow cytometer. For the colorimetric test, alveolar macrophages were incubated with Phorbol 12- miristate-13 acetate and nitroblue tetrazolium, dissolved and analyzed in a spectrophotometer. It was noted that dexamethasone therapeutic dose (0.05 mg/kg) reduced the functions of alveolar macrophages from healthy bovine. This result was observed by both tests with the difference that the flow cytometry assay was more informative for identifying which specific cellular function has been compromised.
Autores principais:Artner,Bruna
Outros Autores:Reck,Angela; Coelho,Alessandra Mayer; Prediger,Julio José; Pontarolo,Desiree Vera; Bertagnon,Heloisa Godoi
Assunto:reactive oxygen species immunity alveolar macrophage
Ano:2018
País:Brasil
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:IBICT
Idioma:inglês
Origem:Oasisbr
Descrição
Resumo:ABSTRACT: Evaluation of alveolar macrophage functions of cattle is an important tool in order to assess whether measures taken during the cattle husbandry can decrease the respiratory tract defense. The aim of this study was to determine whether dexamethasone used at therapeutic dose can affect the oxidative metabolism of alveolar macrophages of cattle. This was evaluated by two tests, the fluorometric and colorimetric. The similarity of the results was studied, using alveolar macrophages of six healthy cattle, obtained from bronchoalveolar lavage on a basal and an immunosuppressant moment after the application of dexamethasone. For the fluorometric test, alveolar macrophages were incubated with Staphylococcus aureus and 2’-7’dichlorohidroflurescein, and analyzed by flow cytometer. For the colorimetric test, alveolar macrophages were incubated with Phorbol 12- miristate-13 acetate and nitroblue tetrazolium, dissolved and analyzed in a spectrophotometer. It was noted that dexamethasone therapeutic dose (0.05 mg/kg) reduced the functions of alveolar macrophages from healthy bovine. This result was observed by both tests with the difference that the flow cytometry assay was more informative for identifying which specific cellular function has been compromised.

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