Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-03-29T14:28:44Z No. of bitstreams: 1 annaelisamartins.pdf: 29809625 bytes, checksum: 335c9c3ea05f09abdd389ff9ab9c36f6 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-04-24T02:48:04Z (GMT) No. of bitstreams: 1 annaelisamartins.pdf: 29809625 bytes, checksum: 335c9c3ea05f09abdd389ff9ab9c36f6 (MD5) / Made available in DSpace on 2016-04-24T02:48:04Z (GMT). No. of bitstreams: 1 annaelisamartins.pdf: 29809625 bytes, checksum: 335c9c3ea05f09abdd389ff9ab9c36f6 (MD5) Previous issue date: 2013-06-27 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A cidade de Juiz de Fora, Minas Gerais, está passando desde o final dos anos 2000, pelo processo de implantação do Parque Científico e Tecnológico de Juiz de Fora e Região, cuja proposta de multifocalidade atenderia às áreas de transferência de tecnologia da UFJF, às respectivas linhas de desenvolvimento tecnológico e de produção científica, em associação com outras duas entidades. Apresentado como um projeto inovador, fundamentado naqueles que vêm sendo instalados em todo o mundo, o objetivo desta dissertação é apresentar um modelo de organização produtiva ocorrida na primeira metade do século XX, na Escola de Engenharia, utilizando-se para isso a pesquisa em fontes primárias, entrevistas, jornais, folhetos de anúncio, propaganda e pequenos informes de circulação locais antigos, além da análise de instrumentos legais municipais, estaduais e federais, como leis, decretos, artigos, estatutos e resoluções para complementação da base teórica. Foi traçada uma linha evolutiva e preenchida lacunas sobre a história da Faculdade de Engenharia, descobrindo-se assim as razões do pioneirismo das associações entre empresas, governos, pesquisadores e universidades, chegando-se à conclusão de que entre os anos de 1940 e 1960 foi desenvolvido um modelo de produção que conseguiu aliar os potenciais industriais com os da prestação de serviços na área educacional, configurando o Partec como o primeiro parque científico e tecnológico da região. / The city of Juiz de Fora, Minas Gerais, is passing since de end of 2000´s, by the process of implementing of the Scientific and Technological Park of Juiz de Fora and Region, whose proposed is to multifocality attend to the areas of technology transfer from UFJF, their lines of technological development and scientific production in association with two other entities. Presented as an innovative project based on those that have been installed worldwide, the goal of this dissertation is to present a model of productive arrangement occurred in the first half of the twentieth century, in the School of Engineering, using it for research, primary sources, interviews, newspapers, advertising brochures, and small reports circulating ancient sites, and analysis of local, state and federal legal, as laws, decrees, articles, and resolutions to complement the theoretical basis. It is hoped that a line can be drawn evolutionary and filled gaps on the history of the Faculty of Engineering, thus discovering the reasons for pioneering associations between companies, governments, researchers and universities, coming to the conclusion that between the years 1940 and 1960 we developed a model of productive arrangement that could combine with the industrial potential of services in education, setting the Partec as the first scientific and technology park in the region.

<p>During bio-technical processing it is important to monitorbiological parameters such as cell growth, viability andproduct formation. Many of the analyses traditionally used areslow to perform and provide only average data for thepopulation. Flow cytometry is a laser-based technique, whichmeasures physical properties of a cell in a flowing stream, ata rate of several thousand cells per second. It offers theprospect of an at-line, multi-parameter analysis of individualmicroorganisms in a population.</p><p>In this project several methods for at-line measurements ofbioprocesses were developed such as protocol's for measuringcell concentration, viability and product formation. Theprimary focus was on prokaryotic organisms (<i>E. coli</i>) but eukaryotic organisms (<i>P. pastoris</i>) were included.</p><p>The possibility to use volumetric cell counting to measurecell concentration (cell number) was evaluated. It was shownthat the method was applicable for high cell density processesof both<i>E. coli</i>and<i>P. pastoris</i>.</p><p>The combination of Bis- (1,3-dibutylbarbituric acid)trimethine oxonol (depolarised membranes) and propidium iodide(loss of membrane integrity) as fluorescent markers was usefulto measure viability at-line of cells in high cell densityprocesses. The protocol was shown to be reproducible for<i>E. coli</i>and<i>P. pastoris</i>.</p><p>The viability staining was used to study the kinetics ofweak organic acids (food preservatives). The protocol provideddata about cell functions such as membrane depolarisation andloss of membrane integrity caused by introducing weak organicacids to shake flask cultures of<i>E. coli.</i></p><p>Labeling inclusion bodies with fluorescent antibodiesprovided a method, which could specifically monitor theincreased accumulation of recombinant promegapoetin proteinwith process time. This technique was further developed forintracellular staining by application of a permeabilising stepbefore labeling with antibodies. Staining of inclusion bodiesdirectly inside permeabilised cells gave information about thedistribution of protein expression in the cell population.</p><p>In conclusion, flow cytometry provides an at-line, singlecell technique for measurement of several biological parametersin bioprocesses.</p><p><b>Key words</b>: flow cytometry, Partec PAS, propidium iodide(PI), bis- (1,3-dibutylbarbituric acid) trimethine oxonol(BOX), Alexa fluor 488, bioprocess,<i>E. coli</i>,<i>P. pastoris</i>, inclusion body, food preservatives,viability, membrane potential</p>

During bio-technical processing it is important to monitorbiological parameters such as cell growth, viability andproduct formation. Many of the analyses traditionally used areslow to perform and provide only average data for thepopulation. Flow cytometry is a laser-based technique, whichmeasures physical properties of a cell in a flowing stream, ata rate of several thousand cells per second. It offers theprospect of an at-line, multi-parameter analysis of individualmicroorganisms in a population. In this project several methods for at-line measurements ofbioprocesses were developed such as protocol's for measuringcell concentration, viability and product formation. Theprimary focus was on prokaryotic organisms (E. coli) but eukaryotic organisms (P. pastoris) were included. The possibility to use volumetric cell counting to measurecell concentration (cell number) was evaluated. It was shownthat the method was applicable for high cell density processesof bothE. coliandP. pastoris. The combination of Bis- (1,3-dibutylbarbituric acid)trimethine oxonol (depolarised membranes) and propidium iodide(loss of membrane integrity) as fluorescent markers was usefulto measure viability at-line of cells in high cell densityprocesses. The protocol was shown to be reproducible forE. coliandP. pastoris. The viability staining was used to study the kinetics ofweak organic acids (food preservatives). The protocol provideddata about cell functions such as membrane depolarisation andloss of membrane integrity caused by introducing weak organicacids to shake flask cultures ofE. coli. Labeling inclusion bodies with fluorescent antibodiesprovided a method, which could specifically monitor theincreased accumulation of recombinant promegapoetin proteinwith process time. This technique was further developed forintracellular staining by application of a permeabilising stepbefore labeling with antibodies. Staining of inclusion bodiesdirectly inside permeabilised cells gave information about thedistribution of protein expression in the cell population. In conclusion, flow cytometry provides an at-line, singlecell technique for measurement of several biological parametersin bioprocesses. Key words: flow cytometry, Partec PAS, propidium iodide(PI), bis- (1,3-dibutylbarbituric acid) trimethine oxonol(BOX), Alexa fluor 488, bioprocess,E. coli,P. pastoris, inclusion body, food preservatives,viability, membrane potential