Submitted by Maike Costa (maiksebas@gmail.com) on 2016-05-05T13:54:43Z No. of bitstreams: 1 arquivo total.pdf: 4535008 bytes, checksum: 35f2a65a445c1e8219acb5615d951d72 (MD5) / Made available in DSpace on 2016-05-05T13:54:43Z (GMT). No. of bitstreams: 1 arquivo total.pdf: 4535008 bytes, checksum: 35f2a65a445c1e8219acb5615d951d72 (MD5) Previous issue date: 2015-07-01 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / In this work were developed and validated two analytical methodologies using solid phase extraction and High Performance Liquid Chromatography (HPLC), for identification and quantification of endocrine disruptors in surface water. A methodology was used to analyze four estrogens: 17 β-estradiol, ethinyl estradiol, estrone and estriol, and the other methodology was used to analyze six phytoestrogens: biochanin A, genistin, genistein, daidzin, daidzein and formononetin. In the sample preparation step we developed an extraction method using solid phase extraction (SPE).The used cartridges were effective in the extraction for both estrogens and phytoestrogens using 1000 ml of river sample, and reached a pre-concentration factor of 2,857 times and recoveries above 90%. The proposed method uses fluorescence detector for estrogens, while the one for the phytoestrogens uses diode array detector. On methods validation were evaluated analytical performances for both methods. The selectivity shows that there was no presence of interfering peaks in both evaluated methods. The generated calibration curves presented good linearity and good correlation coefficients (above 0.99) for all studied analytes. The curves linearity was evaluated by an ANOVA and analysis of graphical residue which showed that the model shows no lack of fit. The limit of detection (LOD) and of quantification (LOQ) for estrogens and phytoestrogens determined in this study were similar to the observed values in other studies and therefore were considered appropriate for the evaluated matrix type. The accuracy of the methods was assessed by the repeatability and intermediate precision, which presented CV <13%. The accuracy was assessed using the recovery test, which was in the range of 72.9 -113%). In the evaluated stability, standard solutions of both methods were stable for a period of 30 days under refrigeration. The results showed that the proposed methods are reliable and have been validated successfully. After validated the methods were applied in the analysis of water samples from the Cuiá River, located in João Pessoa, Paraíba (Brazil), on different collection points, and three sampling campaigns. In all campaigns it was observed the presence of the four estrogens and phytoestrogens six in concentrations ranging from 1.40 to 4754 ng L-1 for estrogens and 3.58 to 2210 μg. L-1 to phytoestrogens in the aquatic studied system. / Neste trabalho foram desenvolvidas e validadas duas metodologias analíticas, utilizando extração em fase sólida e cromatografia líquida de alta eficiência (do inglês: High Performance Liquid Chromatography – HPLC), para identificação e quantificação de interferentes endócrinos em águas superficiais. Uma metodologia foi utilizada para analisar quatro estrógenos: 17 β-estradiol, 17 α-etinilestradiol, estrona e estriol, e a outra metodologia usada para analisar seis fitoestrógenos: biochanin A, genistina, genisteína, daidizina, daidizeína e formononetina. Na etapa de preparo das amostras foi desenvolvido um método de extração empregando extração em fase sólida (SPE). Os cartuchos utilizados foram eficazes na extração dos analitos, tanto dos estrógenos como dos fitoestrógenos, utilizando 1.000 mL de amostra de rio, sendo atingido um fator de pré-concentração de 2.857 vezes e recuperações acima de 90%. O método para os estrógenos utilizou detector de fluorescência, enquanto o método para os fitoestrógenos utilizou o detector de arranjo de diodos. Na validação dos métodos foram avaliadas as figuras de desempenho analítico para ambos os métodos. A avaliação da seletividade demonstrou que não havia a presença de picos interferentes em ambos os métodos avaliados. As curvas analíticas geradas apresentaram linearidade satisfatória e coeficientes de correlação acima de 0,99, para todos os analitos estudados. A linearidade das curvas foi avaliada com os testes de ANOVA e análise dos gráficos de resíduos os quais demonstraram que os modelos não apresentaram falta de ajuste. Os LD’s e LQ’s estimados para os estrógenos mostraram-se similares aos valores apresentados em outros estudos, enquanto que os LD’s e LQ’s estimados para os fitoestrógenos tiveram valores mais altos. No entanto podem ser considerados satisfatórios para o tipo de matriz avaliada. A precisão dos métodos foi avaliada pela repetitividade e pela precisão intermediária, as quais apresentaram CV < 13 %. A exatidão do método foi avaliada através do teste de recuperação, que esteve na faixa de 72,9 a 113 %. Na avaliação da estabilidade, as soluções padrões de ambos os métodos se mostraram estáveis por um período de até 30 dias sob refrigeração. Os resultados demonstraram que os métodos propostos são confiáveis e que foram validados com sucesso. Depois de validados, os métodos foram aplicados nas análises das amostras de água do Rio Cuiá, situado na cidade de João Pessoa, Paraíba (Brasil), em diferentes pontos de coleta e, em três campanhas amostrais. Em todas as campanhas observou-se a ocorrência dos quatro estrógenos e dos seis fitoestrógenos, em concentrações que variaram de 1,40 a 4.754 ng L-1 para os estrógenos e de 3,58 a 2.210 μg L-1 para os fitoestrógenos.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / As substÃncias denominadas desreguladores endÃcrinos (DE) sÃo uma categoria recente de poluentes ambientais que interferem nas funÃÃes do sistema endÃcrino, sendo encontradas no meio ambiente em concentraÃÃes na ordem de &#956;g L-1 e ng L-1. Os efluentes de ETEs sÃo importantes fontes de lanÃamento desses compostos no ambiente aquÃtico uma vez que os processos convencionais de tratamento de Ãgua e esgoto nÃo removem totalmente esses micropoluentes. Neste trabalho avaliou-se a eficiÃncia da argila pilarizada utilizando o diÃxido de estanho como agente pilarizante para remoÃÃo de DE (de diferentes classes) em matrizes ambientais. Foi utilizado o planejamento experimental para avaliaÃÃo dos fatores que afetaram o processo adsortivo. Para etapa de otimizaÃÃo do mÃtodo analÃtico anteriormente validado foi estudada a eficiÃncia de trÃs cartuchos SPE comerciais. Foram obtidas concentraÃÃes afluentes, efluentes e eficiÃncias de remoÃÃo em duas estaÃÃes de tratamento de esgotos que operavam em escala real (UASB + cloraÃÃo). Por fim, foram realizados experimentos de adsorÃÃo de DE em sistemas mono e multicomponentes, assim como de regeneraÃÃo do adsorvente. Os cartuchos Oasis e Strata-X mostraram elevada eficiÃncia e capacidade seletiva, entretanto apenas o cartucho Strata-X foi o selecionado neste trabalho, uma vez que o mesmo mostrou maiores valores de recuperaÃÃo (%) de 50 (Sulfamethoxazol); 83 (Trimetoprima); 171 (Diclofenaco); 183 (Estrona); 14 (17&#946;-estradiol) e 82 (Bisfenol-A). O mÃtodo cromatogrÃfico mostrou-se seletivo para todos os compostos estudados, exceto para o hormÃnio estriol. Os valores de limite de quantificaÃÃo (1,28-29,97 ng L-1) e limite de detecÃÃo (0,42-9,89 ng L-1) obtidos neste trabalho foram considerados baixos. A maioria dos valores de recuperaÃÃo encontrados foi menor que 100%. Foi possÃvel observar que o esgosto afluente Ãs ETEs estudadas ainda apresentam em seus efluentes quantidades considerÃveis de DE, com valores mÃdios efluentes entre 21,94-311,79 ng L-1 para ETE Aracapà e 4,41-123,47 ng L-1 para ETE Mondubim, o que evidencia a necessidade de uma etapa de pÃs-tratamento nas ETEs estudadas. Foram encontradas maiores eficiÃncias de tratamento para os compostos bisfenol A (90,8%) e 17&#945;-etinilestradiol (76,6%) na ETE Mondubim. Jà para os compostos sulfametoxazol e estrona foi observado um incremento nos valores efluentes de 18,9 e 28,6%, respectivamente para a ETE AracapÃ. A argila pilarizada mostrou maior capacidade de adsorÃÃo quando comparada à argila natural, devido ao alargamento do espaÃo interlamelar da argila e aumento no volume de mesoporos como comprovado pelas tÃcnicas de caracterizaÃÃo. ApÃs anÃlise dos resultados do planejamento fatorial foi fixado os valores Ãtimos encontrados das variÃveis no processo para compor os estudos de cinÃtica e isoterma de adsorÃÃo, os quais foram Temperatura â 25 ÂC; AgitaÃÃo â 300 rpm; Massa do adsorvente: 300 mg e pH â 3. O estudo cinÃtico indicou tempo de equilÃbrio de 1 hora, sendo o modelo de pseudo-segunda ordem o que melhor se ajustou aos dados. O mecanismo de difusÃo intrapartÃcula nÃo foi a etapa determinante da velocidade e no processo de transferÃncia de massa. A maioria dos compostos DE estudados obteve maior remoÃÃo em sistema multicomponente quando comparados aos estudos monocomponentes. Os dados experimentais de isoterma de adsorÃÃo melhor se ajustaram ao modelo de Freundlich com exceÃÃo aos referentes aos compostos TMP e BPA. Os testes de regeneraÃÃo do adsorvente indicaram significante reduÃÃo da capacidade de adsorÃÃo da argila pilarizada no 2 ciclo de adsorÃÃo para os compostos TMP e BPA, com reduÃÃo de 99,5 e 79,2 %, respectivamente. Para os compostos E2 e E1, a adsorÃÃo foi afetada apÃs o 3 ciclo. / The substances called endocrine disruptors (ED) are a new category of environmental pollutants that interfere in the functions of the endocrine system, being found in the environment at concentrations in the order ug L-1 and L-1 ng. The WWTP effluent release are important sources of such compounds in the aquatic environment since the conventional methods of water treatment and sewage not completely remove these micropollutants. In this study we assessed the efficiency of pillared clay using tin dioxide as an agent for removing pilarizante DE (of different classes) in environmental matrices. Experimental design was used to evaluate the factors affecting the adsorptive process. For optimization step of the previously validated analytical method it was studied the effectiveness of three commercial SPE cartridges. Tributaries concentrations were obtained, effluents and removal efficiencies of two sewage treatment plants operating in full scale (UASB + chlorination). Finally, DE adsorption experiments were performed in single and multicomponent systems as well as regenerating the adsorbent. The Oasis and Strata-X cartridge efficiency showed high and selective ability, however, only Strata-X cartridge was selected in this study, since it showed higher recovery values (%) 50 (Sulfamethoxazol); 83 (Trimethoprim); 171 (diclofenac); 183 (estrone); 14 (17&#946;-estradiol) and 82 (Bisphenol A). The chromatographic method was selective for all compounds studied, except for the hormone estriol. The quantification limit values (1.28 to 29.97 ng L-1) and detection limit (from 0.42 to 9.89 ng L-1) obtained in this study were low. Most of the recovery values was less than 100%. It was observed that the esgosto tributary of the studied WWTPs still present in their waste considerable amounts of DE, with average values of effluents from 21.94 to 311.79 ng L-1 for ETE Aracapà and 4.41 to 123.47 ng L 1 to ETE Mondubim, which highlights the need for a step of post-treatment in the studied WWTPs. They have found greater efficiencies treatment for compounds bisphenol A (90.8%) and 17&#945;-ethinylestradiol (76.6%) ETE Mondubim. As for the compounds sulfamethoxazole and estrone it was observed an increase in effluent values of 18.9 and 28.6% respectively for the ETE AracapÃ. The pillared clay showed higher adsorption capacity compared to the natural clay, due to enlargement of the interlayer space of the clay and increased volume of mesopores as evidenced by the characterization techniques. After analyzing the results of the factorial design was set optimum values of the variables found in the process to make the studies of kinetics and adsorption isotherm, which was temperature - 25  C; Agitation - 300 rpm; Mass of adsorbent: 300 mg and pH - 3. The kinetic study indicated 1 hour equilibration time, with the model of pseudo-second order which best fit to the data. The intraparticle diffusion mechanism was not the determining step of the speed and mass transfer process. The majority of the DE compounds obtained greater removal in multicomponent system when compared to single-component studies. The experimental data better adsorption isotherm set the model Freundlich except to refer to the TMP and BPA compounds. The sorbent regeneration tests indicated a significant reduction in the adsorption capacity of pillared clay in the 2nd adsorption cycle for TMP and compounds BPA, with a reduction of 99.5 and 79.2%, respectively. For E2 and E1 compounds, adsorption was affected after the 3rd cycle.

Pesticides are among the most important risk factors for the health of workers and the environment. Many are recognized for their actions as endocrine disruptors, leading to changes in thyroid function markers. The aim of the study was to investigate the association between occupational exposure to organophosphate pesticides and thyroid function markers growers. The study enrolled 208 rural workers involved in the orange culture Boquim municipality in April 2015 to April 2016. They answered the questionnaire containing demographic information, health profile and occupational risk. Blood samples were collected for analysis of thyroid function. A large part of the workers live in the countryside 131 (72.0%), belong to social classes D and E (50.8%) and are predominantly male 161 (77.4%). The health profile analysis showed the presence of characteristic symptoms often associated with organophosphate poisoning. Muscle weakness has been reported (45.9%), skin disorders (36.3%), tremor night (17.2%) and seizures (2.2%). With regard to occupational variables, most responders reported having some kind of long, direct or indirect, with the pesticide (92.3%) and were exposed to pesticides to a period longer than five years (66.1%). Biochemical markers of thyroid function were analysed: TSH (thyroid stimulating hormone) with 10.5% of changed values, free T4 (thyroxine) with 41.4% of the reduced values, total T3 (triiodothyronine) with 21.8% of altered values. Altered levels of anti-thyroid peroxidase (anti-TPO) and of anti-thyroglobulin (anti-Tg) were found in 14.7% and 9.3% of cases, respectively. TSH levels and altered T3 predominated in women, and reduced free T4 predominated in men; all had a higher frequency in the fifth decade of life. The presence of positive autoimmunity predominated in women and had a higher frequency in the fourth decade of life for Anti-TPO and fifth for Anti-Tg. Conclusion: The exposure variables to organophosphate pesticides evaluated in this population showed no association with changes in serum TSH, free T4 and total T3 and with overt hypothyroidism diagnosis, subclinical hypothyroidism and hyperthyroidism. / Os agrotóxicos estão entre os mais importantes fatores de risco para a saúde dos trabalhadores e para o meio ambiente. Muitos são reconhecidos por suas ações como disruptores endócrinos, levando a alterações nos marcadores de função tireoidiana. O objetivo do estudo foi investigar a associação entre a exposição ocupacional aos agrotóxicos organofosforados e os marcadores de função tireoidiana de citricultores. Participaram do estudo 208 trabalhadores rurais envolvidos na cultura de laranja do município de Boquim no período de abril 2015 a abril 2016. Responderam o questionário contendo informações sociodemográficas, perfil de saúde e de risco ocupacional. Foram colhidas amostras de sangue dos trabalhadores para a análise da função tireoidiana. A maior parte dos trabalhadores moram na zona rural 131 (72,0 %), pertencem às classes sociais D e E (50,8%) e são predominantemente homens 161 (77,4%). A análise do perfil de saúde demonstrou presença de sintomatologia característica frequentemente associada à intoxicação por organofosforados. Foi relatado fraqueza muscular (45,9%), alteração na pele (36,3%), tremor noturno (17,2%) e convulsões (2,2%). Com relação às variáveis ocupacionais, a maioria dos entrevistados relataram ter algum tipo de contanto, direto ou indireto, com o agrotóxico (92,3%) e estavam expostos aos pesticidas a um período superior a 5 anos (66,1%). Os marcadores bioquímicos da função tireoidiana analisados foram: TSH (hormônio tireoestimulante) com 10,5% dos valores alterados, T4 livre (tiroxina) com 41,4% dos valores reduzidos, T3 total (triiodotironina) com 3,2% de valores alterados. Níveis alterados de anticorpo anti-tireoperoxidase (Anti-TPO) de e anti-tireoglobulina (Anti-Tg) foram encontrados em 14,7% e em 9,3% dos casos, respectivamente. Os níveis de TSH e T3 total alterados predominaram nas mulheres e os de T4 livre reduzidos predominaram nos homens; todos tiveram uma frequência de valores alterados maior na quinta década de vida. Já a presença de autoimunidade positiva predominou nas mulheres e teve uma frequência maior na quarta década de vida para o Anti-TPO e quinta para o Anti-Tg. Conclusão: As variáveis de exposição aos pesticidas organofosforados avaliadas nessa população não demonstraram associação com alterações dos hormônios TSH, T4 livre e T3 Total e com os diagnósticos de hipotireoidismo franco, hipotireoidismo subclínico e hipertireoidismo. / Lagarto, SE

Neste estudo foi realizada a avaliação de seis substâncias sintéticas resultantes de processos industriais e de atividades antropogênicas dietilftalato, dibutilftalato, nonilfenol, pentaclorofenol, bisfenol A e benzo[a]pireno os quais possuem capacidade de interferir negativamente no funcionamento normal do sistema endócrino de animais e de seres humanos (xenoestrógenos), em águas tratada e bruta de quatro municípios do Vale do Paraíba paulista. Para tanto, utilizou-se um método analítico de alta seletividade e sensibilidade, sendo a determinação por cromatografia gasosa acoplada ao detector de espectrometria de massas (GC/MS), precedida da concentração das amostras pelo método SPE (extração em fase sólida). Como não são suficientes apenas boas técnicas analíticas para garantir a qualidade dos dados gerados e a confiabilidade dos resultados, a metodologia desenvolvida foi submetida ao processo de validação, onde foram avaliados os parâmetros: seletividade, especificidade, linearidade, faixa linear de trabalho, limites de detecção e quantificação, precisão, exatidão, recuperação e robustez, além da incerteza na medição. Os resultados demonstraram que o método proposto é adequado para quantificação das amostras do rio Paraíba do Sul, e pela sua aplicação foi constatada a presença de todos os poluentes estudados tanto na água bruta quanto na potável em pelo menos um dos três períodos de amostragem, sendo as maiores concentrações mensuradas nas águas brutas e no período seco. Estes resultados evidenciam que as atividades antrópicas na região influenciam a qualidade da água do recurso hídrico estudado, bem como a qualidade da água de distribuição. / The evaluation of six synthetics substances from industrials process and anthropogenic activities - diethyl phthalate, dibuthyl phthalate, nonylphenol, pentachlorophenol, bisphenol A and benzo[a]pyrene which one can interfer negatively in normal function of the endocrine system from animals and humans (xenoestrogens), was carried out in this study in drinking water and raw water on four cities at region of Paraíba do Sul River. An analytical method with high selectivity and sensitivity, with determination by gas chromatography coupled to mass spectrometry detector (GC/MS), preceded of samples concentration through the SPE technique was used. Good analytical techniques are not enough to ensure quality and reliability of the generated data results. For this, the methodology was submitted to the validation process, where parameters evaluated were: selectivity, specificity, linearity, work range, detection and quantification limits, precision, accuracy, robustness, percentage of recovery and uncertainty in measurement. The validation results showed that proposed method is suitable to quantify Paraíba do Sul river samples, and by its application could observe presence of all studied pollutants in drinking and raw water in one of the three sampling periods at least, and the highest measured concentrations were in raw water at dry periods. These results evidence that the anthropic activities at region are influencing the water quality of the studied water resource, as well as the quality of water for public supply.

Alguns compostos orgânicos como interferentes endócrinos e ativos farmacêuticos estão sendo encontrados em águas superficiais e vêm chamando a atenção de orgãos públicos e da comunidade científica. Dentre esses compostos, foram enfatizados nesse trabalho, o hormônio sintético 17&#945;-etinilestradiol, o surfactante (4-nonilfenol), bisfenol A, o antibiótico &#946;-lactâmico amoxicilina e a cafeína como traçador de atividade antrópica. Estes compostos são continuamente introduzidos em corpos d\'água pelo lançamento de esgoto in natura o que afeta a qualidade da água, a saúde dos ecossistemas e impacta o suprimento de água potável. Considerando a baixa taxa de remoção desses contaminantes pelo tratamento de água atualmente utilizado no Brasil e o estado de degradação dos mananciais de água para abastecimento público, esse trabalho apresenta o desenvolvimento de metodologia analítica para determinação dos compostos citados e a avaliação da eficiência de remoção quando o tratamento convencional de água é aplicado. A determinação foi baseada em métodos já estabelecidos e envolveram as etapas de limpeza e concentração de amostra usando extração em fase sólida (SPE) e análise por cromatografia a líquido de alta eficiência (HPLC) com detector de arranjo de diodos (DAD) e espectrometria de massas. As amostras do Reservatório do Guarapiranga foram utilizadas para ensaios em escala laboratorial que simulavam o tratamento convencional de água em planta real. Os resultados mostraram a adequação dos métodos utilizados para a separação e determinação desses compostos em amostras ambientais. Além disso, apontaram para a necessidade de se avaliar a utilização de metodologias não convencionais que otimizem a remoção pelo tratamento de água atualmente utilizado. / Some organic compounds, such as endocrine disruptors and pharmaceuticals, are being found in the surface water, and are concerning both the public organs and the scientific community. Among these compounds, the synthetic hormone 17&#945;-ethinylestradiol, the surfactant (4-nonylphenol), bisphenol-A, the &#946;-lactam antibiotic amoxicillin and the marker of human activity caffeine were highlighted in the present work. These compounds are continuously drained to the water bodies by sewers, affecting not only the water quality itself, but also the ecosystems, and impacting the drinking water availability. Currently in Brazil, the removal rate of these contaminants is low, whereas the water sources for public supply are highly degraded. In this context, the present work develops an analytical method to detect the aforementioned compounds, and to evaluate the efficiency of its removal by conventional water treatments. Detection was based on pre-established methods and steps involved clean up and samples concentration by solid phase extraction (SPE), high performance liquid chromatography with diode array detection (DAD), and mass spectrometry. Recovery tests were done using environmental samples from Guarapiranga Reservoir (São Paulo, SP). This samples were further used in laboratorial tests, simulating real scale water conventional treatments. Current results revealed the adequacy of the chosen methods in the separation and determination of the abovementioned compounds from natural samples. Also, they highlight the need for the evaluation of non-conventional methodologies that optimize removal by current water treatment.

Many exogenous and endogenous compounds are referred to as endocrine disruptors (EDCs), as they interfere with natural synthesis, signaling and metabolism of endogenous hormones. Common exogenous endocrine disruptors are benzo(a)pyrene (BaP) and 17α-ethinylestradiol (EE2). Endogenous endocrine disruptor 17β-estradiol (E2) is frequently present in the environment as well. In this thesis, the effect of the mentioned EDCs and their combinations on gene and protein expression of CYP1B1, 3A1 and 3A2 in rat liver, kidney and lung was determined. Protein expression was studied using Western blot method and specific antibodies; gene expression was assessed by quantitative PCR. Moreover, the effect of tested EDCs and their combinations on BaP metabolism and CYP3A specific activity (measured as testosterone 6β-hydroxylation) were studied in liver microsomal samples. It was confirmed, that BaP significantly increases CYP1B1 expression in rat liver and lung both alone and together with EE2 or E2. Pretreatment of rat with E2 and BaP increases the ability of BaP to induce CYP1B1 expression. On the contrary, EE2, E2 and their combination decrease the CYP1B1gene expression. The rate of BaP metabolites formed in liver microsomal samples increases in rats pretreated with BaP and its combinations. In liver, there was...

The overall objective of the research presented in this dissertation was to assess exposure to endocrine disrupting chemicals (EDCs), polychlorinated biphenyls (PCBs), phthalates, and bisphenol A (BPA) in the general population and evaluate their associations with adverse reproductive health effects, including cancers, in women. Given the proven contribution of unopposed estrogens to the risk for endometrial neoplasia or breast cancer, renewed health concerns have aroused about estrogen mimicking EDCs found in food, personal care products or as environmental contaminants. Our meta-analysis showed that exposure to estrogen mimicking PCBs increased summary risk of breast cancer and endometriosis. We further evaluated the relationship between endometriosis and breast cancer, and EDCs using a bioinformatics method. Our bioinformatics approach was able to identify genes with the potential to be involved in interaction with PCB, phthalates and BPA that may be important to the development of breast cancer and endometriosis. Therefore, we hypothesized that exposure to EDCs such as PCBs, phthalates, and BPA, results in adverse reproductive health effects in women. Using subject data and biomarkers available from the Center for Disease Controls National Health and Nutrition Examination Survey database we conducted a cross-sectional study of EDCs in relation to self-reported history of endometriosis, uterine leiomyomas, breast cancer, cervical cancer, ovarian cancer, and uterine cancer. Significantly higher body burdens of PCBs were found in women diagnosed with breast cancer, ovarian cancer, and uterine cancer compared to women without cancer. PCB 138 was significantly associated with breast cancer, cervical cancer, and uterine cancer, while PCBs 74 and 118 were significantly associated with ovarian cancer. The sum of dioxin-like PCBs were significantly associated with ovarian cancer (OR of 2.02, 95% CI: 1.06-3.85) and the sum of non-dioxin-like PCBs were significantly associated with uterine cancer (OR of 1.12, 95%CI: 1.03-1.23). Significantly higher body burdens of PCBs were also found in women diagnosed with endometriosis and uterine leiomyomas. Documenting the exposure to EDCs among the general U.S. population, and identifying agents associated with reproductive toxicity have the potential to fill research gaps and facilitate our understanding of the complex role environmental chemicals play in producing toxicity in reproductive organs.

Les perturbateurs endocriniens (PE) posent des risques pour la santé Humaine et pour la faune. L’utilisation de tests biologiques basés sur des mécanismes d’action spécifiques permet de caractériser le potentiel PE des substances chimiques dans le cadre de l'évaluation toxicologique, mais les capacités de biotransformation de ces modèles sont rarement prises en compte. Or, le métabolisme conditionne le devenir des xénobiotiques (détoxication vs. bioactivation) et donc in fine l'activité biologique mesurée. Dans ce travail, le devenir de deux contaminants œstrogéniques émergents (benzophénone-2, bisphénol S) a été comparé dans de nouveaux modèles in vitro et in vivo de poisson zèbre (cellules ZELH-zfERs, larve transgénique cyp19a1b-GFP) et des lignées humaines. Nos résultats démontrent que les modèles de poissons zèbres sont métaboliquement compétents et soulignent leur pertinence dans une approche intégrée in vivo/in vitro pour le criblage de l'activité PE des substances chimiques. / Endocrine disruptors (ED) are potentially harmful for Human beings and for wildlife species. Species-specific mechanism-based screening tests nowadays allow characterizing the ED potency of chemicals in the context of their toxicological assessment. However, the biotransformation capabilities of the biological models used are seldom taken into consideration, despite they ultimately condition the fate of the tested xenobiotics (detoxification vs. bioactivation), and, consequently, the biological response. In this work, we examined the fate of two emerging ED (benzophenone-2, bisphenol S) using novel in vitro and in vivo zebrafish models (ZELH-zfERs cells and cyp19a1b-GFP transgenic larvae) as well as human cell lines. Our results demonstrate that the zebrafish models are metabolically competent, and underline their relevance, accuracy and usefulness in an integrated in vivo/in vitro screening approach designed to assess ED's biological activity

Contexte et objectif : Plusieurs facteurs de risque de cancer ont été identifiés et il a été estimé que plus de 40% des cas dans les pays développés pourraient être évités en modifiant les facteurs de risque connus. L'objectif général de cette thèse était de démontrer que l’intégration de données génomiques et épigénomiques aux données détaillées sur les expositions environnementales et le mode de vie peut être utile pour identifier des biomarqueurs de ces facteurs et contribuer à augmenter notre connaissance de l'étiologie du cancer. Résultats : Dans un premier temps, nous décrivons comment les signatures génomiques et épigénomiques peuvent être utilisées pour identifier des marqueurs d’exposition et déchiffrer l’étiologie du cancer. Ensuite, nous contribuons au débat relatif à l’hypothèse de la chance dans le développement du cancer et démontrons que les mutations induites par le tabagisme sont plus prédictives du risque de cancer que les mutations aléatoires. Nous introduisons un modèle probabiliste pour la simulation de données mutationnelles et comparons la performance des outils d’identification de ces signatures avec des données réelles et simulées. De plus, nous introduisons une nouvelle méthode pour l’identification des signatures mutationnelles. Enfin, nous utilisons les données de méthylation de la cohorte E3N pour étudier le lien entre l'exposition aux retardateurs de flamme bromés et aux composés perfluorés, deux substances classées parmi les perturbateurs endocriniens, et la méthylation de l’ADN sanguin. Globalement, notre étude ne fournit aucune preuve d'altérations globales du méthylome ou d'altérations à l’échelle des CpGs. Cependant, certains résultats suggèrent l’existence d'altérations de la méthylation de gènes impliqués dans des voies biologiques (ex., la réponse aux androgènes) et nécessitent des recherches supplémentaires.Conclusion : Ce travail contribue à la recherche méthodologique portant sur les signatures mutationnelles en introduisant un protocole de mesure de performance et d’identification des signatures mutationnelles pouvant servir de référence à de futures études méthodologiques ou appliquées. Nos recherches sur les signatures mutationnelles et le méthylome démontrent l'utilité de tels outils pour évaluer les expositions et élucider leur rôle dans l'étiologie du cancer. / Context and aim: Several risks factors have been identified for cancer, and it has been estimated that more than 40% of cases in developed countries are preventable through the modulation of known modifiable risk factors. The overall objective of this thesis was to demonstrate that the analysis of genomic and epigenomic data integrated with well-characterised exposure and lifestyle data may be used to identify markers of environmental exposures and lifestyle and may contribute to increase our understanding of cancer aetiology.Results: We first describe how genomic and epigenomic signatures can be used to identify markers of exposure and decipher the aetiology of cancer. Then, we adopt the mutational signatures framework to contribute to the debate about the “bad luck” hypothesis for cancer and demonstrate that tobacco-related mutations are more strongly correlated with cancer risk than random mutations. We introduce a probabilistic model for the simulation of mutational signature data and compare the performance of the available methods for the identification of mutational signatures using both simulated and real data. Additionally, we introduce a new method for the identification of such signatures. Finally, we use methylation array data in an epidemiological study within the E3N cohort to investigate the association between exposure to Brominated Flame Retardants and Per- and polyfluoroalkyl substances, two organic pollutants that are known endocrine disrupting chemicals, and methylation in DNA from blood. Overall, our study does not provide evidence of methylation alterations at the level of the whole genome, in regions or in single CpGs. Suggestive evidence of alterations in the methylation of genes within plausible biological pathways (e.g. androgen response) warrants further investigations. Conclusion: Our work on the methodological aspects of mutational signature research introduces an original framework for measuring the performance of tools for the identification of mutational signatures that may serve as reference for future methodological or applied research. Our applications of both mutational signature and methylome research demonstrate the usefulness of such tools to assess exposures and elucidate their role in cancer aetiology.

Les bisphénols et les phtalates sont des contaminants environnementaux ubiquitaires du fait de leur utilisation massive dans la fabrication de divers produits de consommation. L’exposition à ces molécules est liée à de nombreuses pathologies, dont les cancers hormono-dépendants, mais les mécanismes moléculaires exacts par lesquels elles induisent leurs effets restent largement inconnus.Au cours de ce travail, nous avons étudié l’implication de la sérine thréonine kinase PKD1 dans l’effet de quatre bisphénols : bisphénol A (BPA), bisphénol AF (BPAF), bisphénol F (BPF) et bisphénol S (BPS) et deux phtalates (DEHP et MEHP) sur des cellules du cancer du sein et de la prostate.Nous avons ainsi montré que le BPAF, BPF, BPS, DEHP et MEHP stimulent de manière dose-dépendante la prolifération, la clonogénicité et la croissance indépendante de l’ancrage des cellules MCF-7 et que ces processus sont dépendants de l’expression de PKD1. En revanche, nous avons mis en évidence l’absence d’effet pro-prolifératif du BPA dans trois lignées de cancer de la prostate (LNCaP, PC3 et DU145). D’autre part, nous avons pu identifier que le BPA induit la phosphorylation de PKD1 dans les mitochondries. Enfin, nous avons démontré qu’il existe un effet cumulatif de certaines combinaisons de traitement de bisphénols et de phtalates et mis en évidence que le BPAF induit la transition épithélio mésenchymateuse des cellules MCF-7.Nos résultats permettent ainsi d’identifier PKD1 comme étant une cible moléculaire de certains bisphénols et phtalates dans les cellules MCF-7 fournissant de nouvelles connaissances sur les mécanismes d’action de ces molécules. / Bisphenols and phthalates are ubiquitous environmental contaminants due to their extensive use in the production of a variety of consumer products. Exposure to these molecules is linked to multiple pathologies including hormone-dependent cancers. However, the exact molecular mechanisms by which they exert their effects remain largely unknown.In our study, we analyzed the implication of the serine threonine kinase PKD1 in the effect of four bisphenols : bisphenol A (BPA), bisphenol AF (BPAF), bisphenol F (BPF) and bisphenol S (BPS) and two phthalates (DEHP and MEHP) on breast and prostate cancer cells.We thus showed that BPAF, BPF, BPS, DEHP and MEHP induce, in a dose-dependent manner, the proliferation, clonogenicity and anchorage independent growth of MCF-7 breast cancer cells, via a PKD1-dependent pathway. On the other hand, we showed that BPA does not have a pro-proliferative effect in three prostate cancer cell lines (LNCaP, PC3 and DU145). Furthermore, we determine that BPA induced the phosphorylation of PKD1 into the mitochondria. Finally, we demonstrated that BPAF stimulates epithelial to mesenchymal transition of MCF-7 cells and that co-exposure to combinations of bisphenols and phthalates induces cumulative proliferation effects in MCF-7 cells.Our results characterize PKD1 as a molecular target for bisphenols and phthalates in MCF-7 cells, providing new elements in the knowledge of the mechanisms of action of these molecules.