Challenged by a huge and diverse antigenic stimulus, the intestinal mucosa has developed a unique immune system that mainly functions to maintain tolerance to innocuous antigens while retaining the ability to respond swiftly to pathogenic threats. Central to this specialised immune system are the Intraepithelial Lymphocytes (IELs). These cells are uniquely located between Intestinal Epithelial Cells (IECs) ready to respond to exogenous antigens in the intestinal lumen. The intestinal immune system is constantly influenced, not only by the commensal microbiota, but also by the nutritional status of the host and the availability of certain essential micronutrients that are derived from a healthy-balanced diet. Additionally, age has a significant impact on the efficiency of gut immunity in responding to infectious pathogens, as reflected by the increased burden of gastrointestinal infections at the extremes of age. In this thesis, using the Murine Norovirus (MNV) oral infection model, I aimed to characterize intestinal mucosal antiviral-responses with specific focus on the role of IELs, the impact of aging and the influence of certain micronutrients whose effects are mediated through the Aryl Hydrocarbon Receptor (AhR). Employing different knock-out and adoptive transfer experiments, I concluded that, at least in our experimental conditions and in a viral strain-specific manner, the activated IELs are not essential and may play a minor role in the protective response against MNV infection. This work also demonstrated that various MNV virus strains activate IELs differentially and for the first time (to our knowledge) revealed distinct abilities of these different Norovirus variants to infect IECs. Recognising an impaired response in old (2-year) mice, we were also able to identify a specific defect in the IFN-Lambda response of aged IECs. Furthermore, using the model of MNV infection to investigate the role of AhR signalling, the data I generated suggested a direct link between constitutive AhR signalling and innate interferon-mediated responses. These findings have uncovered a potential preventive/therapeutic targets for enhancing anti-viral responses.

Submitted by RENATA BUZETI GARCIA DE SOUZA null (renatabgarcia90@gmail.com) on 2016-04-11T18:59:55Z No. of bitstreams: 1 PDF DISSERT. Renata Souza-Unesp Bauru.pdf: 5262726 bytes, checksum: f18610acd83b4d02380be38fa0144f6a (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-04-13T12:30:22Z (GMT) No. of bitstreams: 1 souza_rbg_me_bauru.pdf: 5262726 bytes, checksum: f18610acd83b4d02380be38fa0144f6a (MD5) / Made available in DSpace on 2016-04-13T12:30:22Z (GMT). No. of bitstreams: 1 souza_rbg_me_bauru.pdf: 5262726 bytes, checksum: f18610acd83b4d02380be38fa0144f6a (MD5) Previous issue date: 2016-02-29 / Este trabalho tem como objetivo a avaliação da contaminação da água e solo por hidrocarbonetos em dois postos de combustíveis denominados Postos A e B, em dois municípios da região de Avaré (SP). Foram avaliadas as concentrações de contaminantes identificadas nas amostras de água e solo coletadas pelas empresas A e B e, comparadas aos Valores de Intervenção da CETESB para solos e águas subterrâneas. Confirmada a contaminação por hidrocarbonetos em ambos os postos e reclassificadas essas áreas como Áreas Contaminadas (AC), foi avaliada a eficácia das técnicas de remediação executadas no Posto A e, através do comparativo traçado entre 21 casos similares da região de Avaré, traçado um plano de remediação para o Posto B. Foram avaliadas as técnicas praticadas no Posto A, de Bombeamento e Tratamento, Biorremediação e Extração de Vapores do Solo. Os principais resultados mostraram uma significativa redução de BTEX totais entre 74,44% e 97,85%, com exceção apenas do ponto onde encontra-se a bomba de sucção do efluente que apresentou aumento, devido à migração da fase residual. Em relação ao Posto B, o comparativo traçado, resultou em um plano de remediação que conta com a associação das técnicas de Bombeamento e Tratamento e, Biorremediação com o uso de Bioestimulantes, visando a aceleração do processo de remediação, tendo em vista que o único meio de abastecimento de água do Posto B trata-se de poço artesiano. A avaliação das investigações permitiu: a classificação de ambos os postos como AC, a identificação das fontes primárias e secundárias de contaminação, o estudo do fluxo de água subterrânea e a direção e extensão da contaminação ao longo do tempo. Além disso, a metodologia de ações corretivas baseadas no risco (ACBR) permitiu o traçado dos possíveis cenários de risco. O processo de remediação dos postos, continua em operação até que essas concentrações se tornem nulas. / This study aims to evaluate the contamination of water and soil for hydrocarbons in two gas stations called Stations A and B in two cities in the Avare (SP) region. Were evaluated the concentrations of contaminants identified in water and soil samples collected by companies A and B, and compared to CETESB Intervention Values for soil and groundwater. Confirmed the hydrocarbon contamination in both stations and reclassified these areas as Contaminated Areas (CA), it was evaluated the effectiveness of remediation techniques performed in Station A and, through the comparative drawn between 21 similar cases of Avare region, tracing a remediation plan for the Station B. The combination of techniques Pumping and Treatment, Bioremediation and Soil Vapor Extraction used were evaluate regarding the station A. The main results showed a significant reduction of total BTEX between 74.44% and 97.85%, except from the point where was installed the extraction pump which increased the concentration of contaminants because of the movement of the residual phase. About the B Station, the comparative draw resulted in a remediation plan that has the combination of Pumping and Treatment and Bioremediation using Biostimulyators, aiming to accelerate the remediation process, considering that the only way of Station B water supply was a artesian well. The evaluation allowed: the classification of both station as AC, the identification of primary and secondary sources of contamination, the study of groundwater flow, and the direction and extent of the contamination over time and. Besides that, the methodology called corrective actions based in risk (CABR) allowed the possible risk scenarios. The process of remediation stations remains in operation until the concentrations become null.

Submitted by Maike Costa (maiksebas@gmail.com) on 2016-03-09T11:49:24Z No. of bitstreams: 1 arquivototal.pdf: 3160277 bytes, checksum: 9549d58a07164e6f27c72ece8b7ebd35 (MD5) / Made available in DSpace on 2016-03-09T11:49:24Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 3160277 bytes, checksum: 9549d58a07164e6f27c72ece8b7ebd35 (MD5) Previous issue date: 2015-02-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This study aimed to conduct a preliminary and confirmatory assessment of environmental impacts to water resources caused by hydrocarbons originating from car repair shops located in the Distrito dos Mecânicos in Campina Grande - PB. This District was built in the 1980s. Thinking about sustainable practices implies assessing the need for a diagnosis of activities that work with petroleum products and a have high power of contamination of surface and groundwater, such as BTEX (benzene, toluene, ethylbenzene and xylenes), HPAs- Polycyclic Aromatic Hydrocarbons, oils and greases. Called micropollutants, aromatics BTEX and PAHs are the most dangerous among the hydrocarbons because they present high toxicity, even at low concentrations. The analysis of BTEX, PAHs, oils and greases in groundwater were carried out at four places in the Distrito dos Mecânicos, selected from the groundwater flow, and the results were compared with the current legislation for the potability standards and contamination of ecosystems. Samples of surface water were analyzed from two collection points in the Riacho das Piabas, located upstream and downstream of the Distrito dos Mecânicos in order to find a possible influence of the activities performed in the space referred to in the water contamination by hydrocarbons. Through in loco visits it was diagnosed that 91% of the establishment do not have a Oil Water Separator - OWS system and the 9% who have do not perform the proper maintenance. The Companhia de Água e Esgotos da Paraíba also has a OWS system that collects all effluent from the Distrito dos Mecânicos, however, periodic maintenance is not done. During the research we found only the BTEX toluene in groundwater and in surface water. Some PAHs were found only in surface water. The levels of oil and grease in the Riacho das Piabas and groundwater were above those recommended by the CONAMA 430/2011 legislation, which indicates the need for further investigation of the study area and for proposing mitigation measures. / O presente trabalho teve como objetivo realizar uma avaliação preliminar e confirmatória de possíveis impactos ambientais aos recursos hídricos provocados por hidrocarbonetos originados das oficinas mecânicas localizadas no Distrito dos Mecânicos em Campina Grande - PB. Este Distrito foi construído na década de 1980, quando diversas exigências da legislação ambiental ainda não eram previstas. Pensar em práticas sustentáveis implica em avaliar a necessidade de um diagnóstico das atividades que trabalham com produtos derivados do petróleo e que apresentam elevado potencial de contaminação das águas superficiais e subterrâneas, como BTEX (benzeno, tolueno, etilbenzeno e xilenos), hidrocarbonetos policíclicos aromáticos, óleos e graxas. Denominados de micropoluentes, os compostos aromáticos BTEX e HPA são os mais perigosos dentre os hidrocarbonetos por possuírem elevada toxidade, mesmo em baixas concentrações. As análises de BTEX, HPA, óleos e graxas em água subterrânea foram realizadas em quatro pontos do Distrito dos Mecânicos, selecionados a partir do fluxo subterrâneo, e os resultados foram comparados com a legislação vigente para o padrão de potabilidade e contaminação dos ecossistemas. As amostras das águas superficiais foram analisadas a partir de dois pontos de coleta no riacho das Piabas, localizados à montante e à jusante do Distrito dos Mecânicos a fim de verificar uma possível influência das atividades realizadas no referido local na contaminação da água por hidrocarbonetos. Através de visitas in loco foi diagnosticado que 91% das oficinas não possuem instalado o Sistema Separador de Água e Óleo - SSAO e as 9% que possuem não realizam a devida manutenção. A Companhia de Água e Esgotos da Paraíba também possui um sistema SSAO que coleta todo o efluente proveniente do Distrito dos Mecânicos, porém, a manutenção periódica não é realiza. Durante a pesquisa foi encontrado apenas o tolueno em águas superficiais e nas águas subterrâneas. Alguns HPAs foram encontrados apenas nas águas superficiais. Os níveis de óleos e graxas no riacho das Piabas e na água subterrânea estavam acima dos permitidos pela legislação CONAMA 430/2011, o que indica a necessidade de uma investigação mais detalhada da área de estudo e a proposição de medidas mitigadoras.

Made available in DSpace on 2014-12-17T14:10:25Z (GMT). No. of bitstreams: 1 AbinadabeJM_DISSERT.pdf: 2202389 bytes, checksum: c6fb8bcc059666158ed3af8030f88987 (MD5) Previous issue date: 2012-07-19 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / Industrial activities, oil spills and its derivatives, as well as the incomplete combustion of fossil fuels have caused a great accumulation of hydrocarbons in the environment. The number of microorganisms on the planet is estimated at 1030 and prokaryotes the most abundant. They colonized diverse environments for thousands of years, including those considered extreme and represent an untapped source of metabolic and genetic diversity with a large biotechnological potential. It is also known that certain microorganisms have the enzymatic capacity to degrade petroleum hydrocarbons and, in many ecosystems, there is an indigenous community capable of performing this function. The metagenomic has revolutionized the microbiology allowing access uncultured microbial communities, being a powerful tool for elucidation of their ecological functions and metabolic profiles, as well as for identification of new biomolecules. Thus, this study applied metagenomic approaches not only for functional selection of genes involved in biodegradation and emulsification processes of the petroleum-derived hydrocarbons, but also to describe the taxonomic and metabolic composition of two metagenomes from aquatic microbiome. We analyzed 123.116 (365 ? 118 bp) and 127.563 sequences (352 ? 120 bp) of marine and estuarine metagenomes, respectively. Eight clones were found, four involved in the petroleum biodegradation and four were able to emulsify kerosene indicating their abilities in biosurfactants synthesis. Therefore, the metagenomic analyses performed were efficient not only in the search of bioproducts of biotechnological interest and in the analysis of the functional and taxonomic profile of the metagenomes studied as well / Atividades industriais, derramamentos de petr?leo e seus derivados, bem como a combust?o incompleta de combust?veis f?sseis t?m causado um grande ac?mulo de hidrocarbonetos no meio ambiente. O n?mero de microrganismos no planeta ? estimado em 1030, sendo os procariotos os mais abundantes. Eles colonizaram diversos ambientes durante milhares de anos, incluindo aqueles considerados extremos e representam uma fonte inexplorada de diversidade gen?tica e metab?lica com um grande potencial biotecnol?gico. Sabe-se que muitos microrganismos possuem vias metab?licas complexas atuando na biodegrada??o de hidrocarbonetos derivados de petr?leo e, em muitos ecossistemas, existe uma comunidade aut?ctone capaz de realizar essa fun??o. A metagen?mica tem revolucionado a Microbiologia permitindo o acesso ?s comunidades microbianas n?o cultiv?veis, sendo uma potente ferramenta para elucida??o de suas fun??es ecol?gicas, dos perfis metab?licos, bem como para identifica??o de novas biomol?culas. Assim, o presente estudo aplicou abordagens metagen?micas n?o apenas para sele??o funcional de genes envolvidos nos processos de biodegrada??o e biossurfacta??o de hidrocarbonetos derivados do petr?leo, mas tamb?m para descri??o da composi??o taxon?mica e metab?lica de dois metagenomas de microbiota aqu?tica. Foram analisadas 123.116 (365 ? 118 pb) e 127.563 sequ?ncias (352 ? 120 pb) dos metagenomas marinho e estuarino, respectivamente. Oito clones foram encontrados, sendo quatro envolvidos na biodegrada??o de petr?leo e quatro capazes de emulsificar querosene, indicando a habilidade de sintetizar biossurfactantes. Portanto, as an?lises metagen?micas realizadas foram eficientes n?o apenas na busca de bioprodutos de interesse biotecnol?gico como tamb?m na an?lise do perfil funcional e taxon?mico dos metagenomas estudados

Orientador: Chang Hung Kiang / Banca: Walter Malagutti Filho / Banca: Vagner Roberto Elis / Resumo: Visando caracterizar assinaturas geoelétricas em áreas contaminadas por hidrocarbonetos de petróleo, foram realizados levantamentos geofísicos utilizando-se os métodos da eletrorresistividade e análises hidroquímicas em área localizada no Pólo Industrial de Cubatão - SP. Foram realizadas 19 sondagens elétricas verticais (SEVs), 4 imageamentos elétricos 2D e 3 imageamentos elétricos 3D, tendo sido identificadas, por meio destas técnicas, 12, 21 e 9 anomalias, respectivamente. Constatou-se que a presença de anomalias condutivas na zona não saturada coincide com a ocorrência de fase livre nos poços de monitoramento. A identificação de NH4 +, subjacente à ocorrência de fase livre, confirmou a atuação de processo de denitrificação. As baixas concentrações dos íons sulfato (SO4 2-) e ferroso (Fe2+) indicam que os mesmos foram reduzidos biologicamente para sulfeto (S2-) e precipitado como sulfeto de ferro (FeS), respectivamente. Esses resultados permitem concluir que as anomalias condutivas estão diretamente associadas à presença de fase residual de hidrocarbonetos, em processo de biodegradação, na zona não saturada. Desta forma, os métodos de investigação geofísica utilizados permitiram identificar áreas sob influência de hidrocarbonetos de petróleo. / Abstract: Geophysical surveys using electroresistivity methods and hydrochemical analyses were applied aiming the characterization of geoelectrical signatures in hydrocarbon contaminated sites located in the Polo Industrial de Cubatão - SP. There have been accomplished nineteen vertical electric soundings (VESs), four 2D and three 3D electrical imaging, which identified 12, 21 and 9 geophysical anomalies respectively. It was verified that the presence of conductive anomalies within the non-saturated zone coincided with the occurrence of free phase on the monitoring wells. Ammonium (NH4 +) identification, underlying the free phase occurrence, reinforced the activity of denitrifying process. Additionally, low concentration values of sulfate (SO4 2-) and ferrous (Fe2+) ions indicated that they were, respectively, biologically reduced to sulfide (S2-) and precipitated as iron sulfide (FeS). Based on those results, it was concluded that the conductive anomalies detected inside the non-saturated zone were directly associated with the presence of hydrocarbon residual phase, with was undergoing a biodegradation process. Therefore, the geophysical methods applied in this study allowed the identification of sites under petroleum hydrocarbons influence. / Mestre

A Bacia de Santos é uma das principais bacias brasileiras para exploração e produção de hidrocarbonetos, e vem recebendo grandes investimentos em pesquisa nos últimos anos. O estudo das rochas geradoras serve como importante ferramenta para o estudo de modelos de geração, explusão e migração de hidrocarbonetos e na identificação de sistemas petrolíferos. O principal objetivo deste trabalho é a identificação de intervalos e áreas com maior potential para a geração de hidrocarbonetos dentro do pacote sedimentar estudado (Cretáceo-Terciário). As amostras de calha de 6 poços exploratórios, obtidas junto à Agência Nacional de Petróleo, Gás Natural e Biocombustíveis (ANP), foram analisadas através da geoquímica orgânica (Carbono Orgânico Total (COT), Pirólise Rock Eval e análise de Biomarcadores) e da petrologia orgânica (Reflectância da Vitrinita e Palinofácies) e foram interpretadas juntamente com os resultados do estudo de outros 10 poços exploratórios, obtidos pela Geochemical Solutions International cedidos para este trabalho. Esses 16 poços formam um perfil strike SW-NE que recobre praticamente toda a extensão da bacia. Amostras das formações Marambaia, Juréia, Santos, Itajaí-Açu, Itanhaem, Guarujá e Florianópolis foram analisadas, apresentando resultados semelhantes para o tipo de matéria orgânica, sua origem e ambiente deposicional (querogênio tipo II e III, originado pela mistura de material de origem terrestre e marinha depositado em ambiente transicional levemente oxidante). Os perfis analisados identificaram dois intervalos de maior conteúdo de carbono orgânico, maturação e potencial de geração de petróleo, um entre 2500 e 3000 metros de profundidade, que pode estar relacionado a eventos transgressivos do Terciário, e outro entre 4000 a 5000 metros de profundidade, possivelmente relacionado a um importante evento transgressivo de caráter regional ocorrido durante o Turoniano. A matéria orgânica encontrada nos poços situados na região sudoeste do perfil foi a que apresentou as melhores condições para a geração de hidrocarbonetos, tornando-se mais empobrecida na direção nordeste. Com base nos resultados geoquímicos e petrográficos obtidos, a matéria orgânica presente na Formação Itajaí-Açu possui o melhor potencial para geração de hidrocarbonetos, principalmente na porção basal do intervalo, na região sudoeste da bacia. / The Santos Basin is one of the main Brazilian sedimentary basin for hydrocarbon exploration and production and has been the focus of major geological research on hydrocarbon occurrences in recent years. Source rock studies are important tools for construction of hydrocarbon generation, expulsion and migration models of the basin and for identifying petroleum systems. The main objective of the present study is to identify areas and intervals with hydrocarbon generation potential inside the sedimentary sequence studied (Cretaceous-Tertiary). Cutting samples from 6 petroleum exploration wells obtained from the National Agency of Petroleum, Natural Gas and Biofuels (ANP) were analysed by organic geochemical methods (Total Organic Carbon (TOC), Rock Eval Pyrolysis and Biomarkers analysis) and organic petrography (Vitrinite Reflectance and Palynofacies) and were interpreted together with results from organic geochemical studies on samples from 10 addictional petroleum exploratory wells obtained from Geochemical Solution International (GSI), provided for this study. These 16 wells form a SW-NE strike section, that covers the basin almost completely in this direction. The samples from Marambaia, Juréia, Santos, Itajaí-Açu, Itanhaem, Guarujá and Florianópolis formations show similar results in regard to organic matter type, its origin and depositional paleoenvironment (type II-III kerogen, indicating a contribution of terrestrial and marine material, deposited in a transitional and slightly oxic environment). The stratigraphic profiles analysed identify two intervals with relatively high organic matter content, maturation and hydrocarbon generation potential, one between 2500 and 3000 meters depth, perhaps related to a Tertiary transgressive event and the other one between 4000 and 5000 meters depth, possibly related to an important regional transgressive event, which occurred during the Turonian. The organic matter encountered in the well samples located in the southwest of the SW-NE section has the best potential for hydrocarbon generation, whereas the organic matter of well samples located in the northwest direction has less potential to form hydrocarbons. Based on the results of the present study, the organic matter present in the Itajaí-Açu Formation has the best potential for hydrocarbon generation, in particular in the basal part of the formation in the southwest region of the basin.

A Bacia de Santos é uma das principais bacias brasileiras para exploração e produção de hidrocarbonetos, e vem recebendo grandes investimentos em pesquisa nos últimos anos. O estudo das rochas geradoras serve como importante ferramenta para o estudo de modelos de geração, explusão e migração de hidrocarbonetos e na identificação de sistemas petrolíferos. O principal objetivo deste trabalho é a identificação de intervalos e áreas com maior potential para a geração de hidrocarbonetos dentro do pacote sedimentar estudado (Cretáceo-Terciário). As amostras de calha de 6 poços exploratórios, obtidas junto à Agência Nacional de Petróleo, Gás Natural e Biocombustíveis (ANP), foram analisadas através da geoquímica orgânica (Carbono Orgânico Total (COT), Pirólise Rock Eval e análise de Biomarcadores) e da petrologia orgânica (Reflectância da Vitrinita e Palinofácies) e foram interpretadas juntamente com os resultados do estudo de outros 10 poços exploratórios, obtidos pela Geochemical Solutions International cedidos para este trabalho. Esses 16 poços formam um perfil strike SW-NE que recobre praticamente toda a extensão da bacia. Amostras das formações Marambaia, Juréia, Santos, Itajaí-Açu, Itanhaem, Guarujá e Florianópolis foram analisadas, apresentando resultados semelhantes para o tipo de matéria orgânica, sua origem e ambiente deposicional (querogênio tipo II e III, originado pela mistura de material de origem terrestre e marinha depositado em ambiente transicional levemente oxidante). Os perfis analisados identificaram dois intervalos de maior conteúdo de carbono orgânico, maturação e potencial de geração de petróleo, um entre 2500 e 3000 metros de profundidade, que pode estar relacionado a eventos transgressivos do Terciário, e outro entre 4000 a 5000 metros de profundidade, possivelmente relacionado a um importante evento transgressivo de caráter regional ocorrido durante o Turoniano. A matéria orgânica encontrada nos poços situados na região sudoeste do perfil foi a que apresentou as melhores condições para a geração de hidrocarbonetos, tornando-se mais empobrecida na direção nordeste. Com base nos resultados geoquímicos e petrográficos obtidos, a matéria orgânica presente na Formação Itajaí-Açu possui o melhor potencial para geração de hidrocarbonetos, principalmente na porção basal do intervalo, na região sudoeste da bacia. / The Santos Basin is one of the main Brazilian sedimentary basin for hydrocarbon exploration and production and has been the focus of major geological research on hydrocarbon occurrences in recent years. Source rock studies are important tools for construction of hydrocarbon generation, expulsion and migration models of the basin and for identifying petroleum systems. The main objective of the present study is to identify areas and intervals with hydrocarbon generation potential inside the sedimentary sequence studied (Cretaceous-Tertiary). Cutting samples from 6 petroleum exploration wells obtained from the National Agency of Petroleum, Natural Gas and Biofuels (ANP) were analysed by organic geochemical methods (Total Organic Carbon (TOC), Rock Eval Pyrolysis and Biomarkers analysis) and organic petrography (Vitrinite Reflectance and Palynofacies) and were interpreted together with results from organic geochemical studies on samples from 10 addictional petroleum exploratory wells obtained from Geochemical Solution International (GSI), provided for this study. These 16 wells form a SW-NE strike section, that covers the basin almost completely in this direction. The samples from Marambaia, Juréia, Santos, Itajaí-Açu, Itanhaem, Guarujá and Florianópolis formations show similar results in regard to organic matter type, its origin and depositional paleoenvironment (type II-III kerogen, indicating a contribution of terrestrial and marine material, deposited in a transitional and slightly oxic environment). The stratigraphic profiles analysed identify two intervals with relatively high organic matter content, maturation and hydrocarbon generation potential, one between 2500 and 3000 meters depth, perhaps related to a Tertiary transgressive event and the other one between 4000 and 5000 meters depth, possibly related to an important regional transgressive event, which occurred during the Turonian. The organic matter encountered in the well samples located in the southwest of the SW-NE section has the best potential for hydrocarbon generation, whereas the organic matter of well samples located in the northwest direction has less potential to form hydrocarbons. Based on the results of the present study, the organic matter present in the Itajaí-Açu Formation has the best potential for hydrocarbon generation, in particular in the basal part of the formation in the southwest region of the basin.

Investigar, em motores veiculares pesados, alimentados com combustível diesel, a emissão de dioxinas e furanos clorados (PCDD/Fs) e hidrocarbonetos policíclicos aromáticos (PAHs), através da análise de material particulado e gases do sistema de exaustão. Investigar a mutagenicidade do material particulado e gases coletados. Material e Métodos: Os testes de emissão foram feitos em banco de provas dinamométrico de bancada de acordo com Ciclo 13 Pontos de testes, utilizando-se motores Diesel nacionais de aplicação veícular. Um mini-túnel de diluição foi usado para obtenção de amostras de material particulado. As análises de PCDD/Fs e PAHs foram realizadas em laboratório nacional. Para a análise de PCDD/Fs utilizou-se o método USEPA 8290 (1994) e para a análise de PAHs foi utilizado o método USEPA TO 13A (1999). A análise de mutagenicidade foi realizada de acordo com o Teste de Ames (TA98). Resultados: As medições de PCDD/Fs indicaram em geral valores abaixo do limite de detecção do método analítico. Em uma das amostras foi verificado que o OCDD ficou acima do limite de detecção, enquanto em outra isto ocorreu para o OCDF e para o 1,2,3,7,8-PeCDF. Em relação aos PAHs analisados verificou-se a presença dos seguintes compostos: Naftaleno, Acenaftileno, Acenafteno, Fluoreno, Fenantreno, Antraceno, Fluoranteno, Pireno, Benzo[a]antraceno, Criseno, Benzo[b]fluoranteno, Benzo[k]fluoranteno. A média encontrada para 5 amostras analisadas foi de 3.580,5 µg/g de material particulado. Os PAHs encontrados neste trabalho indicam alguma similaridade com a literatura disponível, para valores em % e µg/g de material particulado total coletado. O teste de Ames indicou que o material particulado coletado de motores diesel tem alta atividade mutagênica. Conclusões: A emissão de PCDD/Fs, para o nível de tecnologia dos motores Diesel testados mostrou-se pouco significativa, considerando-se combustível conforme especificado e método e limite de detecção utillizados. Em relação aos PAHs, os resultados indicaram que motores diesel têm uma contribuição importante na formação destes compostos. A alta atividade mutagênica observada mostra que as emissões de veículos Diesel são potencialmente tóxicas. Recomenda-se a continuidade de estudos para definição da regulamentação desses poluentes. / To investigate emission of chlorinated dioxins and furans (PCDD/Fs) and polycyclic aromatic hydrocarbons (PAHs) in heavy duty diesel engines, through the analysis of particulate material and gases in the exhaust.To verify mutagenicity of collected particulate material and gasesous sample extracts. Material and Methods: The engine emissions tests were done in dynamometer test bench according to 13-Mode Cycle, using brazilian engines, vehicle application. A mini dilution tunnel was used to obtain particulate matter and gaseous sample. PCDD/Fs and PAHs analyses were carried out in a certified national laboratory. Dioxins and furans were analyzed according with Method 8290 (USEPA, 1994) and PAHs by Method TO13A (USEPA, 1999). The mutagenicity analyses were done in accordance with Ames test (TA98) Results: The measurements of dioxins and furans in diesel engines operating in controlled conditions in dynamomter bench test, had indicated emissions values in general below the detention limit of the analytical method. In one of the samples it was verified that the OCDD was above the detention limit, while in another one this occurred for OCDF and 1,2,3,7,8-PeCDF. In relation to 16 PAHs it was verified the presence of the following compounds: Naphthalene, Acenaphthylene, Acenaphthene, Fluorene, Phenanthrene, Antracene, Fluorantene, Pyrene, Benzo[a]antracene, Chrysene, Benzo[b]fluoranthene and Benzo[k]fluorantene. The average value of the sum of PAHs found in five samples was 3,580.5 µg.g-1. The PAHs found in this study indicates some similarity with available literature for values in % and µg/g of particulate matter.The Ames test showed that the collected particulate material from diesel engines presents high mutagenic activity. Conclusion: The emission of PCDD/Fs, for the level of technology of the tested Diesel engines revealed little significant, considering the specified fuel, method of analysis and limit of detention used. In relation to the 16 PAHs, the tests results had indicated that diesel engines has an important contribution in the formation of these compounds. The high mutagenic activity observed shows that the emissions of Diesel vehicles are potentially toxic. The continuity of studies for definition of the regulation of these pollutants is recommended.

L'éthanol est converti, à 350°C sous 30 bar et sur des zéolithes protoniques, en un mélange de paraffines légères et d'aromatiques ; produits incorporables dans le pool essence. Cependant, la transformation de EtOH sur zéolithes acides conduit à la formation du coke. Des techniques physiques avancées, en particulier les techniques MALDI et LDI-TOF MS, couplées à la méthode d'analyse qui consiste à récupérer dans un solvant les molécules carbonées après dissolution de la zéolithe dans HF, contribuent à caractériser finement le coke. Sa composition dépend du catalyseur : sur HBEA(11), zéolithe à larges pores, 17 familles ont été détectées contre 4 sur HZSM-5(40) de taille de pore intermédiaire. Sur cette dernière, le coke, composé de polyalkybenzènes / naphtalènes / phénalènes et pyrènes, est localisé à l'intersection des canaux et a une toxicité vis-à-vis des sites acides de Brønsted de 1. En dépit d'un empoisonnement total, cette zéolithe est toujours capable de convertir EtOH, comme MeOH, en hydrocarbures et qui plus est avec les mêmes sélectivités en produits. La transformation de ces deux alcools ne s'explique pas par un mécanisme classique de catalyse acide, mais par un mécanisme concerté radicalaire-acide. La présence d'un inhibiteur de radicaux dans la charge réactionnelle, l'hydroquinone, provoque une désactivation immédiate et une diminution de la concentration des radicaux. La transformation de EtOH et MeOH passe par un intermédiaire réactionnel commun, le carbène :CH2, dont l'oligomérisation radicalaire conduit à la formation d'oléfines. Ces oléfines légères (n-O3-n-O5) sont très réactives et se transforment par catalyse acide (oligomérisation / cyclisation / t / Ethanol is converted into light paraffins and aromatics mixture at 350°C under 30 bar over protonic zeolites. These products can be incorporated in the gasoline pool. Nevertheless, EtOH transformation over acid zeolites leads to the formation of the coke. Advanced physical techniques, in particular MALDI and LDI-TOF MS, were coupled to the coke analysis method. This method consists of the recovery of the carbonaceous molecules in a solvent after zeolite dissolution in hydrofluoric acid solution. This coupling allows characterizing the coke through an extensive way. The coke composition depends on the catalyst morphology: over HBEA(11) zeolite of large pores, 17 families were detected while 4 over HZSM-5(40) of intermediate pore size. Over this latter, the coke, composed of polyalkylbenzenes/naphtalenes/phenalenes and pyrenes, is located in the channels intersections and has toxicity of 1 towards Brønsted acid sites. In spite of a total poisoning, HZSM-5 zeolite is always able to convert EtOH, like MeOH, into hydrocarbons with the same products selectivity. The transformation of the two alcohols cannot be explained by a classical mechanism of acid catalysis, but by a cooperative radical-acid mechanism. The presence of a radical inhibitor in the feed, the hydroquinone, causes an immediate deactivation and a decrease in the concentration of radicals. The transformation of EtOH and MeOH passed by the common reaction intermediate, the :CH2 carbene, which its radical oligomerization leads to the formation of olefins. Olefins (n-O3-n-O5) are very active and can be transformed through acid catalysis (oligomerization/cyclisation/Hydrogen transfer) into aromatics or undergo isomer

Les principaux transporteurs ABC et cytochromes P450 exprimés au niveau des microvaisseaux cérébraux chez l’homme sont l’ABCB1/P-gp, l’ABCG2/BCRP et le CYP1B1. Au niveau des organes périphériques, la régulation de l’expression de ces trois marqueurs fait intervenir des facteurs de transcription et notamment le récepteur aryl hydrocarbon (AhR). Or les transcrits d’AhR sont très exprimés au niveau des microvaisseaux cérébraux humains. Les travaux de cette thèse ont donc dans un premier temps été consacrés à l’étude de l’implication de la voie du récepteur Ah dans la régulation de l’expression de l’ABCB1, de l’ABCG2 et du CYP1B1 au niveau de la barrière hémato-encéphalique. In vivo, nous avons mis en évidence qu’un traitement aigu par la dioxine (ou TCDD), ligand puissant d’AhR, induisait l’expression génique et protéique du Cyp1b1 au niveau des microvaisseaux cérébraux de rats adultes Srague-Dawley. De même, in vitro, l’exposition au TCDD a fortement induit l’expression du CYP1B1 dans la lignée hCMEC/D3, un modèle in vitro de l’endothélium cérébral humain et le recours à la technique de l’ARN interférence nous a permis de démontrer que le récepteur Ah était impliqué dans les effets observés. En revanche, que ce soit in vivo ou in vitro, l’exposition au TCDD n’a entrainé aucune modification significative de l’expression de l’ABCB1 ou de l’ABCG2. Dans un second temps, nous avons étudié l’interrelation entre la voie AhR et les voies de réponse à l’hypoxie cellulaire. Les différentes expériences réalisées sur la lignée cellulaire hCMEC/D3 ont mis en évidence une interaction non réciproque entre ces deux voies de signalisation : en cas d’activation simultanée, la réponse à l’hypoxie abolit la réponse AhR tandis que l’activation de la voie AhR ne modifie pas la réponse adaptative à l’hypoxie. / ABCB1 (P-gp), ABCG2 (BCRP) and CYP1B1 are the main ABC transporters and cytochrome P450 enzymes expressed at the human blood-brain barrier (BBB). In peripheral tissue, expression of these proteins is regulated by transcription factors such as the aryl hydrocarbon receptor (AhR). Interestingly, high levels of AhR mRNA are detected in human brain microvessels. We therefore investigated the potential implication of AhR in the regulation of ABCB1, ABCG2 and CYP1B1 expression. In vivo, a single dose of TCDD, a highly potent AhR ligand, increased Cyp1b1 transcripts and protein expression in rat brain microvessels. Similarly, exposing hCMEC/D3 cells, an in vitro promising model of human BBB, to TCDD induced CYP1B1 expression. Using small interfering RNA, we established AhR involvement in TCDD effects. However, either in vivo or in vitro TCDD treatment had no effect on ABCB1 or ABCG2 expression. Next, we investigated the crosstalk between AhR and hypoxia signalling pathways in case of simultaneous activation. Our experiments revealed that a crosstalk between these two pathways effectively occurred in hCMEC/D3 cells: hypoxia inhibited AhR response but not the reverse.