The aim of this study was to evaluate whether the molecular (kDNA-PCR) and parasitological diagnosis in peripheral blood (PB) could replace the invasive and painful bone marrow collection (BM) in the diagnosis of visceral leishmaniasis (VL). PB from suspected VL patients was evaluated by parasitological and molecular techniques using as the gold standard (GS) a combination of clinical, epidemiological, and immunochromatographic test (PB-rK39) results and parasitological examination of BM. Based on the GS, 38 samples from 32 patients were grouped: Group 1, 20 samples of VL cases, and Group 2, 18 samples of non-VL cases. In order to evaluate the parasitological and molecular techniques in PB, the samples were examined. From Group 1, PB kDNA-PCR was positive in 20 samples and in 19 of 20 in BM kDNA-PCR examination. However, the parasitological examination of buffy coat was insensitive, being able to detect only 4 cases from Group 1. All samples from Group 2 were negative. We concluded that, for the diagnosis of visceral leishmaniasis, the parasitological examination of peripheral blood was not useful; however, molecular diagnosis by kDNA-PCR, performed in peripheral blood, could be useful to replace the parasitological examination of bone marrow.
Dedico este trabalho aos meus queridos e amados pais, Cássia e Manoel, ao meu amado esposo Marcelo, à minha irmã Ramona e sobrinhos. E aos meus avós (in memorian) Marli, Edith e Nino. E à família, razão da minha vida. iv AGRADECIMENTOS À minha orientadora Lúcia Maria Almeida Braz por acreditar no meu potencial, pela intensa dedicação, valiosos ensinamentos, pela amizade e carinho. À Profa. Dra. Thelma Suely Okay pelo apoio desde o começo do projeto, pela participação em todos os aspectos do trabalho e por me acolher em seu laboratório para a realização dos testes moleculares, bem como pela utilização de seus aparelhos, como o Gelbot. Ao querido mestre Prof. Dr. Vicente Amato Neto, pelo constante incentivo e amizade ao longo dos anos. À Profa. Dra. Ester Cerdeira Sabino pelas sugestões e por ter apoiado o desenvolvimento deste projeto. Ao Dr. Manoel Sebastião da Costa Lima Júnior pela colaboração e pelo fornecimento das amostras de pacientes com leishmaniose visceral confirmada procedentes do Mato Grosso do Sul. A FAPESP por financiar projeto (Processo n° 2010/50304-8) que deu origem a este trabalho. À Dra Gilda Del Negrodo Laboratório de Micologia Médicapor me acolher em seu laboratório para a realização de alguns testes moleculares. Ao Dr. Carlos Henrique Valente Moreira-do Instituto Adolfo Lutz-pela realização dos testes estatísticos. À Dra Vera Lucia P. Chioccolado Centro de Parasitologia e Micologia do Instituto Adolfo Lutz-por ceder os oligonucleotídeos iniciadores RV1 e RV2. Ao Dr. José Angelo Lauletta Lindoso e à funcionária Elizabeth, pelo fornecimento de algumas das amostras de pacientes com leishmaniose tegumentar.
and 20g/ml respectively. In(CUR)3 was more effective than other three test agents (IC50: 26g/ml) and the second effective agent against leishmania was Ga(CUR)3 (IC50: 32g/ml). Conclusion: Gallium and indium complexes of curcumin had much lower IC50 values than did their DAC analogs. Based on our study results, In(CUR)3 and Ga(CUR)3 are two important derivatives of the curcumin which need to be more evaluated as potent agents against leishmania Parasites.
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