RESUMO
The causal agent of witches' broom disease, Moniliophthora perniciosa is a hemibiotrophic and endemic fungus of the Amazon basin and the most important cocoa disease in Brazil. The purpose of this study was to analyze the genetic diversity of polysporic isolates of M. perniciosa to evaluate the adaptation of the pathogen from different Brazilian regions and its association with different hosts. Polysporic isolates obtained previously in potato dextrose agar cultures of M. perniciosa from different Brazilian states and different hosts (Theobroma cacao, Solanum cernuum, S. paniculatum, S. lycocarpum, Solanum sp, and others) were analyzed by somatic compatibility grouping where the mycelium interactions were distinguished after 4-8 weeks of confrontation between the different isolates of M. perniciosa based on the precipitation line in the transition zone and by protein electrophoresis through SDS-PAGE. The diversity of polysporic isolates of M. perniciosa was grouped according to geographic proximity and respective hosts. The great genetic diversity of M. perniciosa strains from different Brazilian states and hosts favored adaptation in unusual environments and dissemination at long distances generating new biotypes.
Assuntos
Agaricales/genética , Agaricales/isolamento & purificação , Variação Genética , Esporos Fúngicos/genética , Agaricales/fisiologia , Brasil , Cacau/microbiologia , FilogeniaRESUMO
The purpose of this work was to evaluate biochemical and serological methods to characterize and identify Candida species from the oral cavity. The strains used were five Candida species previously identified: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei, C. tropicalis, and Kluyveromyces marxianus, as a negative control. The analyses were conducted through the SDS-PAGE associated with statistical analysis using software, chromogenic medium, and CHROMagar Candida (CA), as a differential medium for the isolation and presumptive identification of clinically important yeasts and an enzyme-linked immunoabsorbent assay (ELISA), using antisera produced against antigens from two C. albicans strains. This method enabled the screening of the three Candida species: C. albicans, C. tropicalis, and C. krusei, with 100% of specificity. The ELISA using purified immunoglobulin G showed a high level of cross-reaction against protein extracts of Candida species. The SDS-PAGE method allowed the clustering of species-specific isolates using the Simple Matching coefficient, S(SM) = 1.0. The protein profile analysis by SDS-PAGE increases what is known about the taxonomic relationships among oral yeasts. This methodology showed good reproducibility and allows collection of useful information for numerical analysis on information relevant to clinical application, and epidemiological and systematical studies.
Assuntos
Candida albicans/classificação , Candidíase Bucal/microbiologia , Técnicas de Tipagem Micológica/métodos , Animais , Candida albicans/imunologia , Candida albicans/isolamento & purificação , Análise por Conglomerados , Meios de Cultura , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Humanos , Coelhos , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Reactive oxygen species are generated by the rat ovary, and they evoke marked antigonadotropic responses in ovarian cells. Protection against reactive oxygen species is provided by antioxidants such as vitamins C, E, and A, and reduced glutathione (GSH). Our objectives were to establish the ovarian levels of these antioxidants during development and regression of the corpus luteum of the pseudopregnant rat and to determine whether these levels were changed by an acute treatment with either a luteotropic (LH) or luteolytic (prostaglandin [PG] F2 alpha) agent. In addition, we evaluated the extent of oxidative activity in the ovary by determining the level of lipid peroxidation. Follicular development was associated with a significant increase in ovarian levels of vitamin A and GSH, whereas levels of vitamins E and C were unchanged. During the luteal phase, vitamin E levels tended to increase, whereas vitamin A and GSH levels decreased. Luteal regression was associated with a marked increase in ovarian levels of vitamins E and A, whereas GSH levels increased only transiently. Acute treatment with LH in the midluteal phase produced a transient decrease in vitamin C levels that was maximal at 4 h. Luteal vitamin E levels were markedly increased 24 h after LH treatment, whereas vitamin A levels were unchanged, and no evidence of lipid peroxidation was seen. Acute treatment with PGF2 alpha produced a transient decrease in luteal vitamin C levels coincident with transient lipid peroxidation and a sustained fall in serum progesterone levels. Ovarian vitamin A levels were elevated 24 h after PGF2 alpha treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Antioxidantes , Dinoprosta/farmacologia , Glutationa/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Hormônio Luteinizante/farmacologia , Ovário/metabolismo , Vitaminas/metabolismo , Animais , Ácido Ascórbico/metabolismo , Corpo Lúteo/fisiologia , Feminino , Luteólise/fisiologia , Ovário/efeitos dos fármacos , Oxirredução , Pseudogravidez , Ratos , Vitamina A/metabolismo , Vitamina E/metabolismoRESUMO
Este trabalho teve o propósito de avaliar métodos bioquímicos e sorológicos para serem aplicados na caracterização e identificação de linhagens do gênero Candida isoladas da cavidade bucal. As cepas empregadas representam cinco espécies de Candida previamente identificadas: C. albicans, C. guilliermondii, C. parapsilosis, C. krusei e C. tropicalis, utilizando como controle negativo Kluyveromyces marxianus. Foram empregadas as técnicas de gel de poliacrilamida (SDS-PAGE) associado à análise estatística em software, CHROMagar Candida (CA), meio cromogênico diferencial descrito para o isolamento e identificação presuntiva de leveduras de importância clínica e um ensaio de imunoabsorção ligado a enzima (ELISA), utilizando antissoro produzido contra extratos protéicos de uma linhagem-padrão de Candida e um isolado de cavidade oral de C. albicans. O método mostrou-se adequado para a identificação presuntiva de C. albicans, C. tropicalis e C. krusei, com 100% de sensibilidade e especificidade, com base na coloração e textura das colônias. O método de ELISA utilizando imunoglobulinas G purificadas apresentou alto teor de reação cruzada com as outras espécies de Candida estudadas. A análise do perfil protéico por SDS-PAGE permitiu agrupar os isolados da cavidade oral por intermédio do coeficiente "Simple Matching", SSM = 1,0. Os perfis protéicos analisados por SDS-PAGE ampliam os conhecimentos sobre as relaçäes taxonômicas de leveduras isoladas da cavidade oral. Esta metodologia demonstra boa reprodutibilidade e origina informaçäes úteis para aplicação clínica e estudos que envolvem a sistemática e a epidemiologia.
Assuntos
Humanos , Animais , Coelhos , Técnicas de Tipagem Bacteriana , Candida albicans , Candidíase Bucal , Anticorpos Antibacterianos , Compostos Cromogênicos , Análise por Conglomerados , Meios de Cultura , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Verifica mudanças na adaptaçäo e em variáveis psicodinâmicas ao final e no acompanhamento, após 6 meses, de psicoterapias psicodinâmicas breves N=33 (N= 23 mulheres, N=10 homens, idades entre 21 e 60 anos, média 34,3). Apesar da maioria dos sujeitos apresentar adaptaçäo näo eficaz severa no início, as terapias causaram mudanças psicodinâmicas e adaptativas, em parte da amostra, que ainda estavam presentes seis meses após seu término. Mudanças nas estratégias devem ser buscadas, visando os mais comprometidos