ABSTRACT
Bothrops snake venoms have been proved toxic to a variety of cell types, in both in vivo and in vitro models. Studies on the pharmacological actions of Bothrops venoms from Argentina are relatively secarce and the direct action of the crude venoms has not been assessed using cell culture models. In this work, we investigated the cytotoxicity of crude venoms from B. alternatus and B. diporus in a skeletal muscle (C2C12) cell line, which is commonly used as a model for studying the myotoxic action of snake venom. Both venoms (1.25-50 miug/mL) induced an early and significant decrease in cell viability. The cytotoxic concentration 50 (CC50), determined three hours after exposure, revealed that B. diporus venom was significantly more cytotoxic (CC50: 2 miug/mL) than B. aftematus (CC50: 5.8 miug/mL). To investigate the cell death mechanism involved, myoblast cells were examined by phase contrast microscopy and after acridine orange and ethidium bromide fluorescence staining, respectively. Our data clearly demonstrated that an apoptotic mediated this cell line destruction. The current study aimed to provide new information on the citotoxicity meohanisms of Argentine Bothrops snake venoms on a skeletal muscle cell line
Subject(s)
Animals , Male , Female , Crotalid Venoms/toxicity , Apoptosis , Cell Death , Muscle, Skeletal/cytologyABSTRACT
Las especies del género Malassezia han adquirido importancia por su asociación a diversos procesos patológicos. Desde 1996 fueron clasificadas en M. pachydermatis, M. furfur, M. sympodialis, M. slooffae, M. obtusa, M. globosa y restricta. El esquema de identificación basado en las características morfológicas, fisiológicas y bioquímicas no siempre permite distinguir entre algunas de ellas. El objetivo de este trabajo fue poner a punto una metodología rapida, específica y eficaz para la identificación de las especies de este género. Con este fin, basados en la técnica de PCR-REA de Guillot y col., se introdujeron modificaciones en la metodología de extracción y purificación del ADN y en la electroforesis. Las variaciones propuestas en este trabajo confieren mayor practicidad a la técnica y disminuyen los costos. La obtención de resultados definidos permitiría a través de investigaciones taxonómicas y epidemiológicas avanzar en el estudio de la ecología, definir el rol patogénico y aumentar la información epidemiológica del género Malassezia
Subject(s)
Bacterial Typing Techniques , Malassezia , DNA Restriction Enzymes , DNA Restriction-Modification Enzymes , Malassezia , Mycology , Opportunistic Infections , Polymerase Chain ReactionABSTRACT
The genus Malassezia has acquired relevance in the last years for its pathological associations. Since 1996, the genus comprises M. pachydermatis, M. furfur, M. sympodialis, M. slooffiae, M. obtusa, M. globosa and M. restricta. The identification scheme based on morphological, physiological and biochemical characteristics does not resolve ambiguity between some species. The aim of this study was to find out a fast, specific and efficient methodology to distinguish between Malassezia species. Based on a previous technique for polymerase chain reaction-restriction enzyme analysis (PCR-REA), modifications were applied on DNA extraction and purification and the electrophoresis. These changes produced a cheaper, reliable and rapid method as identification procedure with easily defined results. The proposed modifications would allow a better knowledge on ecological and pathogenic roles of Malassezia, studies that have not yet been established.