| spelling |
Susana Johannhttp://lattes.cnpq.br/7244522592099497Betania Barros CotaCarlos Augusto RosaJovita Eugênia Gazzinelli Cruz MadeiraVera Lúcia de AlmeidaFernão Castro BragaDaniel de Assis Santoshttp://lattes.cnpq.br/9643397900925253Cristiane Bigatti Pereira2021-03-30T17:16:03Z2021-03-30T17:16:03Z2013-10-11http://hdl.handle.net/1843/35511A criptococose é uma doença fúngica de caráter oportunista, porém pode acometer pessoas saudáveis. A doença é causada principalmente por Cryptococcus neoformans e Cryptococcus gattii. O número de fármacos antifúngicos para o tratamento da doença é restrito, e por isso buscou-se novos fármacos dentre os fungos endofíticos (FE). A triagem foi realizada por microdiluição com 401 extratos etanólicos de FE isolados de seis plantas medicinais. O único extrato de FE com atividade antifúngica constante foi do FE UFMGCB 2032, identificado como Mycosphaerella spp., oriundo da planta Eugenia aff. bimarginata Berg. coletada no cerrado brasileiro. O fungo foi cultivado em larga escala, o extrato bruto foi particionado com solventes orgânicos e a fração solúvel em acetato de etila fracionada por cromatografia líquida de alta eficiência. Duas substâncias foram isoladas e identificadas por espectrometria de massas e ressonância magnética nuclear. A substância 1 foi isolada pela primeira vez e identificada como o ácido (6E, 12E)-2-amino-3,4-diidróxi-2-(hidróximetil)-14-oxo-icosa- 6,12-dienóico e obteve faixa de concentração inibitória mínima (CIM) entre 0,19-3,12 μg/mL. A substância 2 foi identificada como o ácido (6E)-2-amino-3,4-diidróxi-2-(hidróximetil)-14- oxo-icosa-6-enóico ou miriocina (CIM 0,19-0,78 μg/mL). As substâncias 1 e 2 foram ativas contra os diferentes isolados de Cryptococcus avaliados com CIMs semelhantes as CIMs da anfotericina B (AMB) e menores que as CIMs do fluconazol (FC). Os ensaios de combinação das duas substâncias com a AMB e o FC mostraram 25 % de sinergismo. As duas substâncias foram ativas contra Candida albicans e C. krusei. As substâncias 1 e 2 apresentam baixas taxas de atividade citotóxica contra as células tumorais humanas MCF-7 e TK-10 e baixas porcentagens de inibição contra a forma amastigota-like de Leishmania (Leishmania) amazonensis. A microscopia eletrônica de varredura das leveduras após tratamento com as substâncias mostrou modificações no formato, depressões e enrugamento na superfície e C. gattii também sofreram redução no tamanho e ausência de brotos nas células mãe.Cryptococcosis is an opportunistic fungal disease that may affect healthy people. The disease is mainly caused by C. neoformans and C. gattii yeasts. The options of antifungal drugs available for the treatment are limited, so we search for new antifungal agents among the endophytic fungi (EF). Antifungal trial assays were performed by broth microdilution with 401 ethanolic endophytic fungi extracts isolated from six medicinal plants. The only isolated with constant antifungal activity was the UFMGCB 2032, identified as Mycosphaerella sp., obtained from the medicinal plant E. aff. bimarginata Berg., collected from Brazilian savannah. The fungal was grown on a large scale, the crude extract was submitted to partition with organic solvents, and the ethyl acetate fraction was fractionated by HPLC, which produced two secondary metabolites identified by mass spectra and NMR assays. The (6E, 12E)-2-amino-3,4-dihydroxy-2-(hydroxymethyl)-14-oxo-icosa-6,12-dienoic acid, compound 1, it was isolated by the first time and showed minimal inhibitory concentration (MIC) rate of 0.19-3.12 μg/mL. The second compound was the (6E)-2-amino-3,4-dihydroxy-2- (hydroxymethyl)-14-oxo-icosa-6-enoic acid, compound 2 or myriocin (MIC 0.19-0.78 μg/mL). The two isolated compounds showed antifungal activity against the Cryptococcus species isolates evaluated with MIC values similar to amphotericin B (AMB) MIC, and lower than fluconazole (FC) MIC values. The combined drugs assays indicate 25 % of synergism performed with compounds 1 and 2 combined with AMB and FC. The two compounds also exhibited activity against Candida albicans and C. krusei. The compounds 1 and 2 showed low cytotoxicity rate against MCF-7 and TK-10, and low inhibition percentage against Leishmania (Leishmania) amazonensis. Scanning electronic microscopy of the yeasts treated with isolated compounds permitted the observation of modification in yeasts shape (deformation) and cell wall modifications (depressions and additional wrinkles). Moreover, reduction in the C. gattii size and absence of budding cells yeasts after treatment was seen.CNPq - Conselho Nacional de Desenvolvimento Científico e TecnológicoOutra AgênciaporUniversidade Federal de Minas GeraisPrograma de Pós-Graduação em MicrobiologiaUFMGBrasilICB - DEPARTAMENTO DE MICROBIOLOGIAhttp://creativecommons.org/licenses/by-nc-nd/3.0/pt/info:eu-repo/semantics/openAccessMicrobiologiaCryptococcusCriptococoseFungosCryptococcusCriptococoseFungo endofíticoMycosphaerellaAntifúngicoBioprospecção e caracterização de substâncias antifúngicas capazes de inibir espécies patogênicas de Cryptococcus a partir dos extratos de fungos endofíticos associados a plantas endêmicas de diferentes ecossistemas brasileirosinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisreponame:Repositório Institucional da UFMGinstname:Universidade Federal de Minas Gerais (UFMG)instacron:UFMGORIGINALBigatti.pdfBigatti.pdfapplication/pdf5803929https://repositorio.ufmg.br/bitstream/1843/35511/1/Bigatti.pdfb445bd95b09cb14788f1766f5328c2a8MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811https://repositorio.ufmg.br/bitstream/1843/35511/2/license_rdfcfd6801dba008cb6adbd9838b81582abMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82119https://repositorio.ufmg.br/bitstream/1843/35511/3/license.txt34badce4be7e31e3adb4575ae96af679MD531843/355112021-03-30 14:16:03.411oai:repositorio.ufmg.br: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Repositório InstitucionalPUBhttps://repositorio.ufmg.br/oaiopendoar:2021-03-30T17:16:03Repositório Institucional da UFMG - Universidade Federal de Minas Gerais (UFMG)false
|