Orientadores: Pedro Luiz Rosalen, Marta Cristina Teixeira Duarte / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-16T16:26:49Z (GMT). No. of bitstreams: 1 Queiroz_VanessaCristinaPinPiazentin_M.pdf: 4355114 bytes, checksum: 2e7009da410cba032963dee4cd9aa7ac (MD5) Previous issue date: 2010 / Resumo: A candidíase oral é uma infecção fúngica que tem sido relatada como problema de saúde pública e em decorrência de um crescente índice de cepas resistentes de Candida spp., tem-se aumentado o interesse em produtos naturais como fonte de novos compostos bioativos. Neste sentido, a própolis brasileira apresenta importantes propriedades farmacológicas como antimicrobiana e poderia ser estudada quanto à sua atividade antifúngica. Assim, o objetivo deste trabalho foi avaliar a atividade anti-Candida de três tipos de própolis brasileiras contra formas planctônicas e em biofilme de Candida spp. de maiores incidências em patologias da cavidade oral. Foram utilizados três tipos de própolis selecionadas por suas propriedades antimicrobianas em estudos prévios, os quais estão classificadas na literatura como: tipo 3 (interior do estado do Paraná), tipo 6 (região de mata atlântica do estado da Bahia) e tipo 13 (região de mangue da cidade de Maceió, estado de Alagoas). Para cada tipo de própolis foram obtidos o extrato etanólico (EEP) e suas respectivas frações com diferentes gradientes de polaridade (hexano, diclorometano e acetato de etila). Aos EEP e respectivas frações foram aplicadas as seguintes metodologias para avaliação da atividade antifúngica contra Candida spp. de forma a conduzir um estudo bioguiado: 1- Determinação da Concentração Inibitória Mínima (CIM) e Concentração Fungicida Mínima (CFM) dos EEP dos tipos 3, 6 e 13 e frações hexânica (fr-hex), diclorometânica (fr-dicloro) e acetato de etila (fr-acet) contra as formas planctônicas das leveduras, obtidas de um banco internacional de cepas Centraalbureau voor Schimmelcultures (CBS): Candida albicans (CBS 562), Candida dubliniensis (CBS 7987), Candida glabrata (CBS 07), Candida krusei (CBS 573), Candida parapsilosis (CBS 604), Candida tropicalis (CBS 94), pelo método da microdiluição. 2- Determinação da CIM das própolis 3 e 13 (EEP e respectivas frações diclorometânicas) sobre biofilmes em pré-adesão. 3- Verificação de possíveis alterações ou danos na estrutura celular das leveduras quando submetidas às própolis 3 e 13 (EEP e respectivas frações diclorometânicas) pela Microscopia Eletrônica de Varredura (MEV) em biofilmes em pré-adesão e formados. De maneira geral, as própolis dos tipos 3 e 13 demonstraram forte atividade anti-Candida contra as leveduras analisadas (valores mínimos e máximos de CIM 4,0 e 31,3 µg/mL para EEP da própolis 3 e 1,0 e 7,8 µg/mL para EEP da própolis 13) em relação a do tipo 6 que demonstrou baixa atividade. O EEP tipo 13 e respectiva fr-dicloro se destacaram, por apresentarem efeito fungicida contra todas as cepas analisadas com valores mínimos e máximos de CFM 125 e 500 µg/mL para a fr-dicloro tipo 13. As própolis 3 e 13 (EEP e respectivas fr-dicloro) apresentaram atividade anti-biofilme em pré-adesão com valores máximos de 250 µg/mL para EEP tipo 3 (C. parapsilosis) e 250 µg/mL para fr-dicloro tipo 3 (C. parapsilosis) e CIM 125 µg/mL para EEP tipo 13 (C. tropicalis) e 31,3 µg/mL para fr-dicloro tipo 13 (C. parapsilosis). As imagens de MEV mostram a interferência dos EEP tipos 3 e 13 e respectivas fr-dicloro sobre biofilmes em pré-adesão, entretanto as própolis não foram capazes de desagregar completamente os biofilmes formados de Candida spp se comparadas ao grupo controle (Nistatina). Conclui-se que os três tipos de própolis estudadas apresentaram atividade anti-Candida tanto em organização planctônica quanto em biofilme, entretanto, a fração diclorometânica da própolis do tipo 13 é a fração ativa, que contém importantes compostos efetivos contra Candida spp. e pode ser fonte de novos princípios bioativos para prevenção ou tratamento da candidíase oral, a qual deve ser objeto de estudos futuros / Abstract: Oral candidiasis is a fungal infection that has been reported as a public health problem and due to increased rate of resistant strains of Candida spp. has increased the interest in natural products as sources for new bioactive compounds. Brazilian propolis have important biological properties such as antimicrobial and anticariogenic activity, thus antifungal activity could also be evaluated. The aim of this study was to evaluate the anti-Candida activity of three Brazilian propolis against planktonic and biofilm forms of Candida spp. of higher incidences in oral cavity diseases.Three types of propolis were selected for antimicrobial properties in previous studies, which are classified in the literature as type 3 (within the state of Parana state), type 6 (Atlantic Rainforest of Bahia state) and type 13 (mangrove region, Maceió city, Alagoas state). For each type of propolis were obtained the ethanolic extract (EEP) and respective fractions with different gradient polarity (hexane, dichloromethane and ethyl acetate). The following methodologies were applied on EEP and correspondent fractions to evaluate antifungal activity against Candida spp. by a bioguided study: 1 - Determination of Minimum Inhibitory Concentration (MIC) and Minimum Fungicidal Concentrations (MFC) of EEP types 3, 6 and 13 and hexane (hex-fr), dichloromethane (dichlo-fr) and ethyl acetate (ethyl-fr) against planktonic forms of yeasts from an international bank Centraalbureau voor Schimmelcultures (CBS): Candida albicans (CBS 562), Candida dubliniensis (CBS 7987), Candida glabrata (CBS 07), Candida krusei (CBS 573), Candida parapsilosis (CBS 604), Candida tropicalis (CBS 94) using the microdilution method. 2 - Determination of MIC of propolis 3 and 13 (EEP and dichloromethane fractions) on preformed biofilm. 3 - Verification of possible changes or damages of the yeast cell structure when subjected to 3 and 13 propolis (EEP and dichloromethane fractions) by Scanning Electron Microscopy (SEM) in preformed and mature biofilms. In most cases, propolis types 3 and 13 demonstrated strong anti-Candida activity against the yeasts tested (minimum and maximum MIC values 4.0 and 31.3 µg/mL for EEP type 3 and 1.0 and 7.8 µg/mL for EEP type 13 compared to the low effect of propolis type 6. EEP type 13 and respective dichlo-fr had the best fungicidal effect against all strains (minimum and maximum MFC values 125 and 500 µg/mL). EEP type 3 and 13 and their dichlo-fr presented anti-biofilm activity 250 µg/mL for EEP type 3 (C. parapsilosis), 250 µg/mL for dichlo-fr type 3 (C. parapsilosis) and (maximum MIC 125 µg/mL for EEP type 13 (C. tropicalis), 31.3 µg/mL for dichlo-fr type 13 (C. parapsilosis). SEM images show the influence of propolis samples on preformed biofilms, however these samples were not capable of completely disrupting the mature biofilms, if compared to control (Nistatin). Thus, we concluded that the three types of propolis studied presented anti-Candida activity both in organization and in planktonic biofilm, however, the dichloromethane fraction of propolis type 13 is the active fraction, which contains important compounds effective against Candida spp. and could be a source of new bioactive principles for prevention or alternative treatment of oral candidiasis, which should be the object of future studies / Mestrado / Farmacologia, Anestesiologia e Terapeutica / Mestre em Odontologia

No description available.

Aisla, identifica y determina el perfil de sensibilidad in vitro frente a antifúngicos de especies del género Candida aisladas de flujo vaginal. Se aislaron 60 cepas de Candida de 300 pacientes con vulvovaginitis atendidos por consulta externa en el servicio de ginecología del Hospital “Dos de Mayo” de la ciudad de Lima – Perú. Los estudios de aislamiento, identificación y determinación de la Concentración Mínima Inhibitoria (CMI) fueron realizados en el Instituto de Investigación de Química Biológica, Microbiología y Biotecnología “Marco Antonio Garrido Malo” de la facultad de Farmacia y Bioquímica de la Universidad Nacional Mayor de San Marcos. Las especies aisladas fueron Candida albicans 45 (75%), Candida glabrata 7 (11,6%), Candida tropicalis 6 (10%), Candida krusei 1 (1,7%) y Candida parapsilosis 1 (1,7%). Dichas especies se identificaron según procedimientos estándares, se utilizó CHROMagar Candida® y se realizó la prueba del tubo germinal para la identificación presuntiva; la identificación definitiva se realizó mediante asimilación (Auxonograma) y fermentación de carbohidratos (Zimograma). Los valores de CMI obtenidos in vitro mediante la técnica de microdilución en caldo, estandarizada por el National Committee for Clinical Laboratory Standards (NCCLS, M27-A2), oscilaron en el rango de 0.5 a 16 µg/mL para fluconazol, de 0.03 a 0.125 µg/mL para ketoconazol y de 0.03 a 0.25 µg/mL para la anfotericina B. Estos resultados indican que las especies de Candida estudiadas, son significativamente sensibles frente a los azoles (fluconazol y ketoconazol) y a la anfotericina B. / Tesis

Objetivo: Describir las características epidemiológicas de la candidemia nosocomial. Materiales y métodos: Se realizó un estudio descriptivo, serie de casos; en el Instituto Especializado de Salud del Niño, durante el período de Diciembre del 2001 a Diciembre del 2002. El acopio de información se realizó mediante la Ficha de recolección de datos. Los datos obtenidos se procesaron por medio del programa SPSS 10.0 para Windows. Resultados: Se identificó 17 casos de candidemia nosocomial, distribuidos en los servicios de Cardiología ( 35.3%), Infectología (17.6%) y UCI (17.6%) principalmente. El 41.2% de los casos eran mayores de 28 días y menores de un año, seguido en frecuencia por el grupo neonatal con 23.5%. El promedio de la estancia hospitalaria prolongada, fue de 30.7 días. Los factores de riesgo intrínsecos más frecuentemente observados fueron la desnutrición ( 58.8%), la colonización por Cándida ( 35.3%) y la infección por VIH-SIDA. Los extrínsecos más frecuentes fueron la terapia antibiótica y la línea endovenosa periférica ( 100% ), la ventilación mecánica ( 52.9%), la cirugía previa y la estancia en UCI. El 41.2% de lo hemocultivos positivos a Cándida, se tipificaron como albicans. Tres casos en tratamiento con Fluconazol resultaron con candidemia. La tasa de letalidad de la candidemia nosocomial resultó en 64.71%. Conclusiones: La mayor frecuencia de casos se observó en los servicios de Cardiología, UCI e Infectología; con un mayor compromiso en la población menor de un año. Los factores de riesgo intrínsecos más frecuentemente observados fueron la desnutrición y la colonización por Cándida; entre los extrínsecos se observó a la terapia antibiótica, la línea endovenosa periférica y la ventilación mecánica. El 41.2% de los casos fue ocasionado por la especie albicans. La tasa de letalidad fue de 64.71%.

During the infection process Candida albicans has to respond to various stresses imposed by host environment including oxidative and osmolarity stress generated by phagocytic cells such as macrophages and neutrophils. Exposure to caspofungin and other antifungal antibiotics also imposes stress on the C. albicans cell wall. These various stress responses are orchestrated through the activation of multiple stress pathways including the cAMP-PKA, several MAPK pathways and the Ca2+-calcineurin pathway which influence cell wall shape and composition. Such changes were predicted to influence recognition of C. albicans by innate immune cells. During my Ph.D. studies I primarily investigated the effect of the activation or inhibition of these pathways on the interaction with the innate immune cells by examining phagocytosis, the cytokine profile induced by mononuclear and polynuclear cells of the innate immune system. I found that the activation and inhibition of these pathways plays an important role in remodeling of cell wall and hence the immunological profile. Inactivation of cAMP, Calcium signaling pathway by the deletion of TPK1 and CNA1 resulted in marked reduction in pro-inflammatory cytokine production. Inactivation of MAPK pathway by deletion of HOG1 altered major pro-inflammatory cytokine secretion. Cytokine production was also affected by exposure of C. albicans signaling mutants to Calcofluor White, caspofungin, oxidative and osmotic inducing stresses. Cytokine stimulation was also affected by deletion of URA3, exposure of C. albicans to rifmapicin and antimycin A. These results suggest that stress signaling pathways act to regulate collateral changes in the cell wall, which in turn affects the immune reactivity. Pro and anti-inflammatory cytokine and antifungal profiles of Candida haemulonii was also found to be highly variable. Thus regulation and exposure to different microenvironments significantly modifies immunological signature of fungal cells, suggesting that responses to local stresses make the fungal cell surface a moving target for immunological surveillance.

Background Oral candidiasis is the most common infection of the oral mucosa of HIV-seropositive patients, although its frequency is rapidly decreasing with the advent of highly active antiretroviral therapy (HAART). Many questions regarding its complex pathogenesis remain unanswered. The diagnosis is usually established with non-invasive techniques such as mucosal smears. Oral lesions of HIV-associated Kaposi sarcoma (HIV-KS) are routinely biopsied and frequently show secondary infection with Candida albicans or other Candida species. Aims and objectives The aim of this investigation was to determine the frequency and histomorphology of secondary Candidal infection of the surface epithelium of oral HIV-associated KS lesions (HIV-KS), which are routinely biopsied in HIV infected patients. Materials and methods Haematoxylin and eosin (HE), and Periodic Acid-Schiff (PAS) stains of 133 cases of oral Kaposi sarcoma diagnosed between the period 2003 and 2007 within the Division of Oral Pathology were examined histologically for intensity and morphology of Candidal colonisation, depth of invasion, number of organisms, epithelial reactions and associated inflammatory response. The depth of Candidal invasion and severity of infection were correlated with the available CD4 T cell counts of HIV seropositive patients at the time of biopsy. Results Almost forty one percent (40.62%) of all oral HIV-KS cases were secondarily infected with Candida species. The intensity varied from an isolated single pseudohyphus to matted colonies of vegetative yeasts and psuedohyphae. Whilst in most cases the organisms did not invade beyond the parakeratin layer, pseudohyphae were noted extending into the stratum spinosum in 2 cases, and a single case showed a pseudohyphus within the lamina propria. A further 2 cases showed pseudohyphae growing in the pyogenic membrane. Neutrophilic permeation of the epithelium and Munro micro-abscess formation, features commonly associated with Candidal infection, were frequently present even in the absence of Candidal infection. Candidal organisms were often present in the absence of inflammation. Conclusion Oral lesions of HIV-KS are commonly secondarily infected with large numbers of Candidal organisms. The morphological characteristics of secondary Candidal infection within the surface epithelium of HIV-KS lesions suggest an altered pathogenetic pathway. Further studies are necessary in this regard.

Thesis (Ph. D.)--University of Hong Kong, 2006. / Title proper from title frame. Also available in printed format.

Objetivo: Describir las características epidemiológicas de la candidemia nosocomial. Materiales y métodos: Se realizó un estudio descriptivo, serie de casos; en el Instituto Especializado de Salud del Niño, durante el período de Diciembre del 2001 a Diciembre del 2002. El acopio de información se realizó mediante la Ficha de recolección de datos. Los datos obtenidos se procesaron por medio del programa SPSS 10.0 para Windows. Resultados: Se identificó 17 casos de candidemia nosocomial, distribuidos en los servicios de Cardiología ( 35.3%), Infectología (17.6%) y UCI (17.6%) principalmente. El 41.2% de los casos eran mayores de 28 días y menores de un año, seguido en frecuencia por el grupo neonatal con 23.5%. El promedio de la estancia hospitalaria prolongada, fue de 30.7 días. Los factores de riesgo intrínsecos más frecuentemente observados fueron la desnutrición ( 58.8%), la colonización por Cándida ( 35.3%) y la infección por VIH-SIDA. Los extrínsecos más frecuentes fueron la terapia antibiótica y la línea endovenosa periférica ( 100% ), la ventilación mecánica ( 52.9%), la cirugía previa y la estancia en UCI. El 41.2% de lo hemocultivos positivos a Cándida, se tipificaron como albicans. Tres casos en tratamiento con Fluconazol resultaron con candidemia. La tasa de letalidad de la candidemia nosocomial resultó en 64.71%. Conclusiones: La mayor frecuencia de casos se observó en los servicios de Cardiología, UCI e Infectología; con un mayor compromiso en la población menor de un año. Los factores de riesgo intrínsecos más frecuentemente observados fueron la desnutrición y la colonización por Cándida; entre los extrínsecos se observó a la terapia antibiótica, la línea endovenosa periférica y la ventilación mecánica. El 41.2% de los casos fue ocasionado por la especie albicans. La tasa de letalidad fue de 64.71%.

published_or_final_version / Pathology / Doctoral / Doctor of Philosophy

Darbe analizuotas makšties mikrofloros pakitimai moterų, sergančių skirtingų stadijų endometrioze. Tirtos 46 sergančios moterys. Remiantis mūsų atliktais tyrimais kandidozė nustatyta moterims sergančioms I stadijos endometrioze 3 (23 proc.), sergančioms II stadijos endometrioze 19 (76 proc.) ir sergančioms III stadijos endometrioze 2 (50 proc.). / In this work the chantes of vaginal mikroflora among women suffering from different stages of endometriosis are analized. 46 women have been investigated. Having analysed the research data it has been noticed that Candidiasis is more among women suffering from the virst stage of endometriosis 3 (23%), the second stage of endometriosis 19 (76%) or the third stage 2 (50%).