Incluir como ag?ncias financiadoras: Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (CAPES), Financiadora de Estudos e Projetos (FINEP) e Funda??o de Amparo ? Pesquisa do Estado de Minas Gerais (FAPEMIG). / Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2018-02-07T21:39:03Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) marcelo_henrique_fernandes_ottoni.pdf: 3524322 bytes, checksum: c7675d522b34cfb2455e18dbca1699da (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2018-03-09T19:17:22Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) marcelo_henrique_fernandes_ottoni.pdf: 3524322 bytes, checksum: c7675d522b34cfb2455e18dbca1699da (MD5) / Made available in DSpace on 2018-03-09T19:17:22Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) marcelo_henrique_fernandes_ottoni.pdf: 3524322 bytes, checksum: c7675d522b34cfb2455e18dbca1699da (MD5) Previous issue date: 2017 / Conselho Nacional de Pesquisas (CNPq) / A presen?a de fluoresc?ncia interferente em uma dada amostra ? considerada como um importante problema em m?todos fluorim?tricos, devido ? poss?vel sobreposi??o espectral entre ela e a emiss?o fluorescente de sondas. Por isso, ? importante conhecer se h? fluoresc?ncia interferente em uma dada amostra e subst?ncias de interesse nas condi??es de trabalho pretendidas. No presente estudo, foi avaliada a presen?a de fluoresc?ncia interferente em linf?citos humanos ap?s o tratamento com extratos de tr?s diferentes plantas medicinais, sendo estas, objeto de estudo do nosso grupo de pesquisas. Os extratos s?o: o extrato etan?lico das partes a?reas de Ageratum fastigiatum, extrato etan?lico das partes a?reas de Eriosema campestre e o extrato etan?lico do caule de Pseudobrickellia brasiliensis. Foi coletado o sangue de tr?s volunt?rios para a separa??o das c?lulas mononucleares de sangue perif?rico, para a confec??o de culturas in vitro em meio RPMI devidamente suplementado. Foram feitas uma cultura controle n?o-tratada (CON), culturas tratadas com cada extrato em tr?s concentra??es diferentes, al?m de uma cultura de c?lulas tratadas com dimetilsulf?xido (DMSO), solvente usado na solubiliza??o dos extratos vegetais. As culturas celulares foram incubadas por 24 horas a 37 ?C e 5% de CO2. Ap?s esse per?odo, as c?lulas foram lavadas e avaliadas por citometria de fluxo ou por microscopia confocal. A presen?a de fluoresc?ncia interferente foi determinada com base em histogramas de intensidade de fluoresc?ncia feitos para oito intervalos de comprimento de onda distintos, tendo sido feitas an?lises quali e quantitativas dos dados. A fluoresc?ncia dos extratos vegetais e DMSO foram avaliadas isoladamente por fluorimetria, usando-se as mesmas concentra??es utilizadas para as culturas celulares. Atrav?s da citometria de fluxo, foi identificado que o tratamento de linf?citos com qualquer um dos tr?s extratos de plantas levou ao aparecimento de fluoresc?ncia interferente detect?vel em v?rias faixas de comprimento de onda. Culturas tratadas com DMSO n?o apresentaram fluoresc?ncia interferente. Pela fluorimetria foi visto que os extratos n?o emitem fluoresc?ncia, o que sugere que a fluoresc?ncia interferente foi induzida nas c?lulas ap?s intera??es entre elas e os extratos. Este estudo levanta precau??es que visam avaliar a poss?vel presen?a de fluoresc?ncia interferente em condi??es de trabalho, possibilitando evitar esse vi?s e aumentar a confiabilidade dos resultados. / Disserta??o (Mestrado) ? Programa de P?s-gradua??o em Ci?ncias Farmac?uticas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2017. / The presence of interfering fluorescence in a given sample is considered as an important problem on fluorometric methods due to the possible spectral overlap between it and the fluorescent emission of probes. Therefore, it is important to know if there is interfering fluorescence in a given sample, as well as, in substances of interest, in the desired working conditions. In the present study, it was evaluated the presence of interfering fluorescence in human lymphocytes after the treatment with extracts from three different medicinal plants, being such plants the object of study of our research group. The extracts are: the ethanolic extract from the aerial parts of Ageratum fastigiatum, ethanolic extract from the aerial parts of Eriosema campestre and the ethanolic extract from the stem of Pseudobrickellia brasiliensis. Blood was collected from three volunteers to get the peripheral blood mononuclear cells, for in vitro culture preparation in properly supplemented RPMI medium. It was made an untreated control culture (CON), cultures treated with each extract at three different concentrations, and a culture of cells treated with dimethylsulfoxide (DMSO), the solvent used in the plant extracts solubilization. Cell cultures were incubated for 24 hours at 37?C and 5% CO2. After that, cells were washed and evaluated by flow cytometry or by confocal microscopy. The presence of interfering fluorescence was determined based on making fluorescence intensity histograms at eight wavelength ranges for each cell culture. Qualitative and quantitative data analyzes were performed with those graphs. Plant extracts and DMSO fluorescences were evaluated by fluorometry, using the same concentrations used for the cell cultures. In flow cytometry results, it was identified that the treatment of lymphocytes with any of the three plant extracts led to the appearance of interfering fluorescence detectable in several wavelength ranges. Cultures treated with DMSO showed no interfering fluorescence. By fluorometry it was seen that the extracts are not fluorescent, which suggests that the interfering fluorescence was induced in the cells by interactions between them and the extracts. This study raises precautions to evaluate the possible presence of interfering fluorescence in working conditions, making possible to avoid this bias and increase the results reliability.