Memoria CD.indd - ISHAM

4-31Transcriptional profile of Paracoccidioides brasiliensis induced by oenothein B, anantifungal agent from Brazilian Savannah plant Eugenia unifloraP. F. Zambuzzi 1 , Rezende, R. V. 1 , Borges, C. L. 1 ,Ferri, P. H. 2 , Santos, S. C. 2 , Soares, C. M. A. 1 and Pereira, M 11 Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal de Goiás, Goiania-GO-Brazil. 2 Laboratório deBioatividade Molecular, Instituto de Química, Universidade Federal de Goiás, Goiania-GO-Brazil. e-mail: mani@icb.ufg.brParacoccidioides brasiliensis is a soil-borne fungus and the causative agent of paracoccidioidomycosis, asystemic mycosis that cause chronic and progressive infection. The long-time treatment, the high toxicity of thedrugs, and the appearance of resistant or multi-resistant strains have imposed the need for a permanent searchand development of new therapeutic approaches. A large variety of compounds extracted from plants has beenused to the discovery of new antimicrobial agents. The active compound oenothein B, purified from leaves ofEugenia uniflora, a Brazilian Savannah plant, has been evaluated on the growth, viability and gene expressionof P. brasiliensis. That compound interferes with yeast cell morphology and inhibits glucana synthase transcriptsaccumulate; suggesting that oenothein B can be a good candidate to antifungal agent. Aiming elucidate themechanism of action of oenothein B on P. brasiliensis, was realized Representational Difference Analysis (RDA).P. brasiliensis yeast cells were grown on MMcM (Mc Veigh & Morton) medium in the presence (tester) and in theabsence (driver) of oenothein B for 90 min, at 37 o C. After extraction of total RNA, the cDNA was obtained andused to RDA experiments according with modified protocol previously described by Pastorian et al. (2000). RDAexperiment originated 280 ESTs that were successfully sequenced and originated 28 contig and 24 singlets. Usingthe BLATX program, was possible classified the ESTs in agreement with the functions. The analyses indicated thepresence of transcripts with functions related with cell wall and membrane, elongation and transcription factors andhypothetic proteins. The elucidation of the action mechanism of oenothein B will facilitate the design and synthesisof related compounds with enhanced pharmacological profiles.4-32Transcriptional profile of murine macrophages infected with pathogenic fungus HistoplasmacapsulatumSimoneide.S.S 1 , Passos-Silva. D. G. 2 ., Teixeira. S.M.R 2 , Junta, C. 3,4 , Medeiros. A. I. 6 , Silva. C. L. 5 , Faccioli. L. H. 6 , Passos.G.A.S. 3,4 , Felipe. M.S.S. 11Departamento de Biologia Celular, Universidade de Brasília, Brasília, DF, Brazil. 2 Departamento de Bioquímica e Imunologia, UFMG,Belo Horizonte, MG, Brazil. 3 Departamento de Genética, USP, Ribeirão Preto, SP, Brazil. 4 Faculdade de Odontologia, USP, RibeirãoPreto, SP, Brazil. 5 Faculdade de Medicina de Ribeirão Preto, USP, Ribeirão Preto 14040-900, SP, Brazil. 6 Faculdade de CiênciasFarmacêuticas, USP, Ribeirão Preto, SP, Brazile-mail: simoneide.silva@gmail.comHistoplasma capsulatum is the thermo-dimorphic fungus that causes histoplasmosis. The yeast form of P.brasiliensis acts as a facultative intracellular pathogen, being able to survive and replicate within macrophages asa probable mechanism to evade the immune system. In the present work the kinetic profile of murine macrophagesinfected with H. capsulatum was investigated. A clinical isolate of the fungus was used for macrophage infection.Total RNA from infected and non-infected macrophages at 6, 24 and 48 hours was extracted with the TRIzol ®reagent (Invitrogen, USA) and hybridized against glass arrays. After normalization, the Significance Analysis ofMicroarrays software (SAM- http://www-stat.standord.edu/) was used to assess the significant variations in geneexpression between experimental and control conditions. Genes were considered to be modulated if they crossedtwo statistical thresholds (q value