JOSÉ MARIA SOUZA
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Resumo IPEN-doc 24556 Inorganic chemical characterization of the soil and sediment from Taiaçupeba Reservoir, São Paulo2017 - LEONARDO, L.; SOUZA, J.M.; DAMATTO, S.R.; SURKOV, A.M.; MADUAR, M.F.; SILVA, A.R.Taiaçupeba reservoir, located in the state of São Paulo, Brazil, belongs to Producer System of Alto Tietê (Sistema Produtor Alto Tietê –SPAT) and it is responsible for water supply for about 3.1 million of people. The water quality of a reservoir became very important in the last decades due to the increase of environmental degradation of the soil and its several uses. Trace and major elements can accumulate in high levels in soil and sediment, and the study of its concentration can indicate if the ecosystem is polluted. Therefore, the study of soil profiles and sediment cores are an important tool for the understanding of geophysical and geochemical aspects of aquatic ecosystems. The objective of this work was to present the elements As, Ba, Br, Ca, Ce, Co, Cr, Cs, Eu, Fe, Hf, K, La, Lu, Na, Nd, Rb, Sb, Sc, Ta, Tb, Th, U, Yb and Zn concentrations using Instrumental Neutron Activation Analysis (INAA) in four soil profiles and four sediment cores collected in the influence area of Taiaçupeba reservoir. To verify if these elements could be enriched and the ecosystem polluted the Enrichment Factor and the Geoacumulation Index were also used. Soil profiles were collected in trenches up to 2m deep and sampled every 5 cm; in the laboratory the samples were dried at room temperature. The sediment samples were collected with a manual PVC sampler sliced every 2 cm or according to their textural characteristics, and dried in an oven at 50 C; the length of the sediment cores ranged from 42 cm to 61 cm. After drying, soil and sediment samples were sieved in a 2 mm mesh and packed in polyethylene bottles for INAA. The elements Na, As and Sb presented the highest values for both soil and sediment samples, implying in continuous, regular and spatial monitoring of the reservoir. One sediment core presented a very high concentration of the element Zn when compared to the values of the Upper Continental Crust, indicating a possible anthropic contribution from the region’s industries and classifying the reservoir as extremely polluted in relation to this element. In the evaluation of the enrichment factor it was concluded that the soil presented very high enrichment for the elements Na, As and Sb and the sediment presented very severe enrichment for the elements Na, As and Zn.Artigo IPEN-doc 24079 Natural radionuclides from U-238 and TH-232 series and inorganic chemical characterization of soil profiles and sediment cores of the Taiaçupeba Reservoir, São Paulo, Brazil2017 - SOUZA, J.M.; DAMATTO, S.R.; LEONARDO, L.; SURKOV, A.M.; SILVA, A.R.; MADUAR, M.F.; GONÇALVES, P.N.Taiaçupeba reservoir, located in the state of São Paulo, Brazil, belongs to Producer System of Alto Tietê (Sistema Produtor Alto Tietê) and it is responsible for water supply for about 3.1million of people. The water quality of a reservoir is very important, but this is reduced by the increase of environmental degradation of the soil around the reservoir and its different uses. The study of soil profiles and sediment cores is an important tool for understanding the geophysical and geochemical aspects of an aquatic ecosystem. The objective of this work was to present the natural radionuclides 238U, 226Ra, 210Pb, 232Th, 228Th, 228Ra and 40K activity concentrations and also the inorganic chemical characterization of four soil profiles and four sediment cores collected in the area of influence area of Taiaçupeba reservoir. The analytical techniques, gamma spectrometry and instrumental neutron activation analysis were used in the determination. In the soil profiles the highest activity concentrations were obtained for the radionuclides 40K and 228Th and the lowest for 210Pb; in the sediment cores the highest activity concentrations were obtained for the radionuclide 210Pb and the lowest for 226Ra and 228Ra. For the inorganic chemical characterization the highest values obtained were for Na, As and Sb; in a sediment core a very high concentration was obtained for the element Zn indicating a probable accumulation of this element inside the reservoir; enrichment factor was used to evaluate a possible anthropic contamination in the soil and sediment at the margins of Taiacupeba reservoir.Resumo IPEN-doc 21725 Preliminary results fo sup(238)U, sup(232)Th and trace elements determined in soil and sediment profiles collected in Ponte Nova Reservoir, São Paulo, Brazil2015 - DAMATTO, S.R.; LEONARDO, L.; MADUAR, M.F.; SILVA, A.R.; SOUZA, J.M.; MAZZILLI, B.P.Resumo IPEN-doc 11434 The use of bacterioophage lambda Psub(L) as a constitutive promoter2006 - HELLER, S.R.; OLIVEIRA, T.L.; UEDA, E.K.; SOUZA, J.M.; OZAKI, N.A.; RIBELA, M.T.C.P.; BARTOLINI, P.; SOARES, C.R.S.Resumo IPEN-doc 15978 Two-step purification of mouse prolactin expressed in the periplasmic space of Escherichia coli2010 - SUZUKI, MIRIAM F.; ARTHUSO, FERNANDA dos S.; SOUZA, JOSE M.; OLIVEIRA, JOAO E.; OLIVEIRA, TAIS L.; OLIVEIRA, NELIO A. de J.; GOULART, HERBERT R.; CAPONE, MARCOS V.N.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.Resumo IPEN-doc 07731 Utilization of reversed-phase high performance liquid chromatography (RP-HPLC) for the analysis of recombinant thyrotropin directly in CHO cell conditioned medium2001 - OLIVEIRA, J.E.; MENDONCA, F.; PERONI, C.N.; SOUZA, J.M.; BARTOLINI, P.; RIBELA, M.T.C.P.Resumo IPEN-doc 11011 Reversed-phase high performance liquid chromatography (RP-HPLC) analysis and hydrophobicity studies of recombinant human pituitary hormones synthesized in E. coli and CHO cells2005 - RIBELA, M.T.C.P.; CARVALHO, C.M.; HELLER, S.R.; LOUREIRO, R.F.; OLIVEIRA, J.E.; OZAKI, N.A.; PERONI, C.N.; SOARES, C.R.J.; SOUZA, J.M.; UEDA, E.K.; BARTOLINI, P.The synthesis and laboratory production of human growth hormone (hGH) and prolactin (hPRL) have been carried out in genetically modified E. coli, while those of thyroid stimulating hormone (hTSH) and of two analogs antagonists of hPRL (G129RhPRL and S179D-hPRL) in stably transfected CHO cells. Human follicle stimulating hormone (hFSH) and luteinizing hormone (hLH) have not been synthesized yet in our laboratory but their HPLC analytical methodologies are under development. For the purpose of studying and improving synthesis and bioreaction yields and, at the same time, planning and following all subsequent purification steps, novel RP-HPLC methods have been set up for each hormone. For hGH and hPRL, isocratic RP-HPLC methods have been developed that can qualitatively and quantitatively analyze the two hormones directly in osmotic shock fluids, already during fermentation. For hTSH, hFSH and hLH, RP-HPLC gradient elutions have been set for the analysis of these hormones in their purified form and in CHO conditioned medium. For these three glycoproteins hydrophobicities have been compared and the following order established: hLH>hTSH>hFSH. An analogous hydrophobicity index has been also determined for hPRL and its analogs, being G129R-hPRL>hPRL>S179D-hPRL. Still concerning hFSH, for the first time it has been possible to optimize RP-HPLC elution conditions that are able to preserve its undissociated heterodimeric structure. Thanks to this tool it was thus possible to carry out a comparative study on pituitary, urinary and CHO-derived hFSH preparations, revealing differences that are probably due to the carbohydrate moiety, as already observed for hTSH. Classical highperformance size exclusion chromatography (HPSEC) together with MALDI-TOF-MS analysis was also employed along with these studies, to complement physico-chemical characterization of our proteins of interest.Resumo IPEN-doc 06990 Hollow fiber bioreactors for hormone production in CHO cells2000 - SOARES, C.R.J.; PERONI, C.N.; MORGANTI, L.; AFFONSO, R.; ARKATEN, R.R.; SOUZA, J.M.; BARTOLINI, P.Artigo IPEN-doc 14342 Produção em escala laboratorial de prolactina humana em células CHO adaptadas para o crescimento em suspensão2009 - ARTHUSO, FERNANDA dos S.; SUZUKI, MIRIAM F.; GOULART, HERBERT R.; SOUZA, JOSE M.; OLIVEIRA, JOAO E.; DAMIANI, RENATA; OLIVEIRA, TAIS L.; CAPONE, MARCOS V.N.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.O Grupo de Hormônios do Centro de Biotecnologia do IPEN tem desenvolvido várias linhagens de células de ovário de hamster chinês (CHO) modificadas geneticamente e comprovadamente eficientes na expressão de proteínas heterólogas, dentre elas a prolactina humana (hPRL). No entanto, todas as nossas linhagens são cultivadas aderidas e dependentes da presença de soro fetal bovino (SFB) no meiode cultivo para um crescimento eficiente. Nesse trabalho foi realizada a adaptação da nossa linhagem produtora de hPRL para o crescimento em suspensão e na ausência de SFB. As células em suspensão apresentam um grande interesse industrial-farmacêutico, tanto pela facilidade de cultivo e ampliação de escala, como pela produtividade volumétrica. Os estudos para adaptação dessas células foram realizados com base no protocolo descrito por Sinacore e colaboradores (2000). Desta forma foi realizada uma produção laboratorial de hPRL em frascos "spinner" por 30 dias consecutivos, assim com, a sua purificação e caracterização.Artigo IPEN-doc 14128 Expressão de prolactina de camundongos no espaço periplastico de Escherichia coli2009 - SUZUKI, MIRIAM F.; ARTHUSO, FERNANDA dos S.; SOUZA, JOSE M.; OLIVEIRA, JOAO E.; OLIVEIRA, TAIS L.; OLIVEIRA, NELIO A. de J.; BARTOLINI, PAOLO; SOARES, CARLOS R.J.A prolactina é um neuro hormônio que faz parte da superfamília das citocinas e está envolvida em mais de 300 processos biológicos. Considerando-se a diferença de 41% encontrada na seqüência de aminoácidos da prolactina de camundongo em relação à humana, os experimentos que utilizam a prolactina humana em modelos animais (ratos ou camundongos), podem sofrer interferência de forma decisiva na interpretação dos resultados em estudos de doenças crônicas como o lúpus eritematoso sistêmico e a artrite reumatóide Nesse trabalho introduzimos o cDNA da prolactina de camundongo (mPRL) junto com o gene do peptídio sinalizador DsbA em um cassete de expressão bacteriano desenvolvido em nosso laboratório que utiliza o promotor λPL. Esse plasmídeo foi utilizado para transformar diferentes cepas de Escherichia coli e o clone com expressão periplásmica mais eficiente foi selecionado para estudo de expressão, purificação e caracterização físico-química e biológica da mPRL.