In this thesis I explore sunscreen use-practices, imaginaries, and material flows. I aim to understand how young people in the globally-immersed UK use sunscreens and how they makesense of them. Specifically, I follow Donna Haraway’s practice of implosion, employing theconcepts of care and toxicity to explore how sunscreens are employed to protect particularbodily ideals, aspects of embodied subjectivities and material-discursive boundaries. In following these various forms of protection, I ask what normative discourses, ecologies andbodies are, in turn, made vulnerable. The main corpus of my study comes from interviewconversations with eight people. Their stories highlight how sunscreens mediate understandings of skin, places, bodies, and social relations. I analyse sunscreen advertising from 1979to 2019 to investigate harmonies and conflicts between the cultural imaginaries curated bysunscreen companies and the stories which emerged from these interview conversations. Ialso put these materials into conversation with sunscreen discourses from the biological sciences, asking how these cultural works infuse sunscreen practices and material-semiotics.This thesis is an exploratory journey which encounters tanning, ageing, beauty, ‘race’,smells, places, intimacies, sun-burns, skin cancers, bodily boundaries, ‘Endocrine Disruptors’, toxic ecologies, and emplaced knowledges. It traverses disciplinary boundaries, following attitudes from the feminist and environmental post-humanities in bringing togetherapproaches, methods and theorising from many varied fields of scholarship usually locatedin the humanities, social sciences, and ‘natural’ sciences. As such, this thesis is a wide-ranging, thick description of how sunscreens are used in desires for protection and what is contingently and consequently made vulnerable.

Cette thèse porte sur l’évaluation de l’impact de certains composés environnementaux sur la fécondité féminine, tel que mesuré par le délai de conception (« time to pregnancy » en anglais, ou TTP). Cette recherche a été réalisée dans le cadre de l’Étude mère-enfant sur les composés chimiques de l’environnement (MIREC), une cohorte de grossesse de 2001 femmes recrutées durant le premier trimestre dans dix villes canadiennes de 2008 à 2011. Les données des questionnaires et les échantillons biologiques ont servi à évaluer l’effet de deux groupes de composés : les persistants [composés perfluorés – perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA) et perfluorohexane sulfonate (PFHxS)] et les non persistants (bisphénol A, triclosan et phtalates). Cette thèse comprend également une analyse du potentiel du ratio index-annulaire (2D:4D) comme mesure de sensibilité endocrinienne. À ce jour, des mesures anthropométriques ont été collectées sur environ 800 mères-enfants dans le cadre de l’Étude mère-enfant sur les composés chimiques de l’environnement : biomonitoring et neurodéveloppement à la petite enfance (MIREC CD Plus), un suivi de la cohorte MIREC portant sur la croissance et le développement des enfants jusqu’à 5 ans. Sur l’ensemble, les résultats de cette thèse permettent d’étoffer les preuves concernant les effets adverses potentiels de plusieurs contaminants environnementaux sur la fécondité féminine, telle que mesurée par le TTP. Dans le premier article, nous avons montré une association entre les PFOA et les PFHxS et une baisse de fécondité, ce que d’autres recherches avaient déjà révélé. Dans le deuxième article, nous avons évalué l’effet du triclosan sur le TTP, ce qui n’avait jamais été examiné, pour montrer un délai plus élevé chez les femmes du quartile supérieur d’exposition. De plus, nos résultats sont en accord avec ceux de la seule étude ayant évalué l’effet du Bisphénol A sur la fécondité féminine, qui n’avait pas détecté d’effet. Finalement, nos données semblent indiquer une association entre l’exposition des femmes aux phtalates et un TTP plus court, mais ces résultats ne sont pas statistiquement significatifs. En ce qui a trait au potentiel du ratio index-annuaire (2D:4D) pour mesurer la sensibilité endocrinienne chez les femmes, nos données ne permettent pas d’établir une association entre ce ratio et le TTP. Pour ce qui est des enfants, nous n’avons pas trouvé d’effet adverse entre le tabagisme de la mère durant la grossesse et leur ratio 2D:4D. Par conséquent, nos données ne semblent pas justifier l’utilisation du ratio 2D:4D pour mesurer la sensibilité endocrinienne en lien avec le potentiel reproducteur (basé sur le TTP) ou l’exposition des enfants au tabac durant le premier trimestre de grossesse. / In this thesis, we aimed to evaluate the impact of selected environmental compounds on female fecundity as measured by time to pregnancy (TTP). This research was conducted in the framework of the Maternal-Infant Research on Environmental Chemicals (MIREC) study, a pregnancy cohort of 2001 women recruited during the first trimester of pregnancy in ten cities across Canada between 2008 and 2011. Questionnaire data and biological samples were analyzed to assess the effect of two groups of compounds: persistent [perfluorinated compounds - perfluorooctane sulfonate (PFOS), perfluorooctanoate (PFOA), and perfluorohexane sulfonate (PFHxS)-] and nonpersistent chemicals (Bisphenol A, Triclosan, and phthalates). In addition, this thesis aimed to examine the potential of the second to fourth finger digit ratio (2D:4D) as a sensitive-endocrine endpoint. To this end, anthropometric measurements were obtained in about 800 children and their mothers during the Early Childhood Biomonitoring and Neurodevelopment Study (MIREC-CD Plus), a MIREC follow-up conducted to measure growth and development up to age five. Overall, the results of this thesis have contributed to the evidence regarding the potential adverse effect of several environmental contaminants (ECs) on female fecundity as measured by TTP. In the first article, we found that PFOA and PFHxS were associated with diminished fecundity, supporting previous evidence that suggested a similar effect. In the second article, we assessed for the first time the effect of Triclosan on TTP, presenting evidence of delayed fecundity at the highest quartile of exposure. In addition, our findings agreed with those of the only study that has assessed the effect of Bisphenol A on female fecundity, and which showed no effect. Finally, we found some indication that female exposure to phthalates might be associated with a shorter TTP, although this finding did not reach statistical significance. With regard to the potential of the digit length ratio (2D:4D) as an endocrine-sensitive endpoint in women, our data do not support a strong association between 2D:4D and TTP. In children, we did not find an adverse impact of maternal smoking during pregnancy on children’s 2D:4D. Thus, our data do not support evidence to suggest that 2D:4D could be used as a potential reproductive endocrine-sensitive endpoint in women as measured by TTP, and in their offspring as measured by exposure to maternal smoking during the first trimester of pregnancy.

Hormonaly active compounds in wastewaters represent nowdays a serious problem. Proceses currently used in watewater treatment plants (WWTP) are unefficient in removing these compounds from contaminated wastewaters. The compounds are supposed to sorb onto solid sludge elements and sediments. In this work seven endocrine disruptors were detected in the sludge samples from WWTPs. A new sensitive method for detection of seven selected endocrine disruptors (4-nonylphenol, bisphenol A, estriol, 17β-estradiol, estrone, 17α- ethynylestradiol, irgasan) was developed. The method is based on accelerated solvent extraction (ASE), gel permeation chromatography (GPC) and solid phased extraction. For final extract analysis, gas chromatography coupled with mass spectrometry (GC/MS) was used. The efficiency of this method was tested using artificially contaminated sludge and the method was used to analyse real samples from several WWTPs in Czech Republic. The effect of sludge age on detection of individual analytes was assessed as well. The concentrations of endocrine disruptors measured in the samples reached up to 1 µg/g. The results are comparable or higher then those reported in other works and they show the necessity of further research on endocrine disruptors in the environment.

La différenciation sexuelle des Isopodes dépend d'une hormone sexuelle protéique, l'hormone androgène (HA), caractéristique des Malacostracés. Cet Insulin-Like Peptide suffit à induire par sa présence la différenciation mâle de ces Crustacés. Nous avons identifié in silico le transporteur circulant de l'HA, l'IGFBP-rP1, chez de nombreuses espèces d'Isopodes ainsi qu'à l'échelle des Crustacés. De la même façon, nous avons identifié deux récepteurs transmembranaires, l'IR1 et l'IR2, issus d'une duplication de gène spécifique des Malacostracés. Les patrons d'expression de ces gènes ont été étudiés sur notre espèce modèle, Armadillidium vulgare. Av-IGFBP-rP1 et Av-IR1 sont exprimés de manière ubiquiste et tout au long du développement. Av-IR2 est aussi exprimé à chaque stade de la différenciation mais ce transcrit est quasi-spécifique des glandes androgènes et ovaires. Une approche par ARNi a confirmé l'implication de ces trois protéines dans la voie de signalisation de l'HA. En effet, l'inhibition de l'HA, Av-IGFBP-rP1 et Av-IR1 provoquent l'hypertrophie des glandes androgènes, suggérant leur implication dans une boucle de rétro-contrôle de l'HA. L'inhibition de Av-IR2 semble seulement provoquer la différenciation d'ouvertures génitales femelles. Ces phénotypes sont comparables à ceux des intersexués mâles induits par la bactérie féminisante endogène Wolbachia. Nous montrons cependant que la bactérie altère seulement l'expression de l'HA et pas celle des récepteurs. Enfin, nous avons testé l'effet du bisphénol A mais nous n'observons pas d'altération de la différenciation sexuelle des larves lors d'expositions à ce perturbateur endocrinien exogène. / Sexual differentiation in Isopods relies on a proteinaceous sex hormone called androgenic hormone (AH), specific to Malacostracans. This Insulin-Like Peptide induces male differentiation by its mere presence in these Crustaceans. We identified in silico the circulating carrier of the AH, called IGFBP-rP1, in many Isopod species, but also on the crustacean scale. Similarly, we identified two transmembrane receptors, IR1 and IR2, coming from a gene duplication specific to Malacostracans. The expression patterns of these genes were investigated in our model species, Armadillidium vulgare. Av-IGFBP-rP1 and Av-IR1 are broadly expressed in the animal and throughout development. Av-IR2 is also expressed at each developmental stage but this transcript is almost specific to androgenic glands and ovaries. An RNAi approach has confirmed the implication of these three proteins in the AH signalling pathway. Indeed, the inhibition of AH, Av-IGFBP-rP1 and Av-IR1 induces androgenic gland hypertrophy, suggesting their implication in an AH feedback loop. Av-IR2 inhibition seems to provoke the differentiation of female genital apertures only. These phenotypes are similar to those of male intersexes induced by the endogenous feminizing bacterium Wolbachia. Yet, we show that the bacterium alters the expression of the AH only and not the one of its receptors. Finally, we have tested the effect of bisphenol A but we observe no alteration of the sexual differentiation in larvae upon exposition to this exogenous endocrine disruptor.

Hormonell wirksame Chemikalien, wie Pflanzenschutzmittel oder Pharmaka gelangen durch Abwässer in die Umwelt und akkumulieren vor allem in Oberflächengewässern. Ein erhöhtes Augenmerk liegt auf Substanzen, die durch (anti)androgene und (anti)östrogene Wirkungsweise die Reproduktion von Tieren und Menschen beeinträchtigen. Bei den bisherigen Nachweismethoden für diese Stoffe handelt es sich um invasive Methoden, die das Töten der Tiere beinhalten. Diesen Methoden mangelt es jedoch an der nötigen Sensitivität, um umweltrelevante Konzentrationen der endokrinen Disruptoren (EDs) nach Kurzzeitexposition nachweisen zu können, sowie am Vermögen, alle vier Wirkmechanismen (androgen, antiandrogen, östrogen und antiöstrogen) mit einer einzelnen Testmethode feststellen und unterscheiden zu können. In dieser Studie wurde deshalb mit Hilfe männlicher Afrikanischer Krallenfrösche (Xenopus laevis) eine Testmethode entwickelt, bei der die Frösche verschiedenen (anti)androgenen und (anti)östrogenen EDs ausgesetzt wurden und ihr Rufverhalten untersucht wurde. Diese nicht-invasive Methode erwies sich als schnell und höchst sensitiv. Zudem war es erstmals möglich, die vier verschiedenen Wirkmechanismen allein anhand veränderter Ruftypen und Rufparameter zu bestimmen und zu unterscheiden. Darüber hinaus konnte gezeigt werden, dass bei Anwendung dieser Methode die Möglichkeit besteht, die Versuchstiere in weiteren Tests wiederzuverwenden, da die Rufparameter nach einer expositionsfreien Zeit von sechs Wochen wieder Kontrollwerte erreichten. Zusammengefasst kann die hier vorgestellte verhaltensphysiologische und damit nicht-invasive Methode als Biomarker für den Nachweis von (anti)androgenen und (anti)östrogenen EDs verwendet werden. Ferner zeigt die hohe Sensitivität des Tests, sowie die Möglichkeit der vollautomatischen Analyse enormer Datenmengen, dass dieser schnelle Verhaltenstest ein großes Potential hat, ein sensitiver, standardisierter und nicht-invasiver Biomarker zu werden. / Endocrine disrupting compounds (EDCs), such as herbicides, pesticides or pharmaceuticals enter the environment via sewage effluents and especially accumulate in surface waters. Research efforts so far mainly focused on EDCs with (anti)androgenic and (anti)estrogenic modes of action (MOAs), which can interfere with reproductive biology of vertebrates. To date, biomarkers for the assessment of such compounds are invasive techniques, which are not sensitive enough to detect EDCs after short-term exposures and which cannot distinguish between the four MOAs. Hence, in this study a non-invasive method for the assessment of EDCs was developed using male African clawed frogs (Xenopus laevis) as model species. Frogs were exposed to individual (anti)androgenic and (anti)estrogenic EDCs in the surrounding water and their calling behavior was analyzed. This non-invasive method turned out to be a fast and highly sensitive biomarker for the detection of (anti)androgenic and (anti)estrogenic EDCs. Moreover, this method was able to differentiate between the four different MOAs solely by determining affected parameters of the calling behavior. It was also shown that by using this method, it might be possible to reuse already tested experimental animals, because the measured affected parameters were reversed after a period of six weeks under control conditions. Taken together the here established non-invasive behavioral method can be used as biomarker for the detection of (anti)androgenic and (anti)estrogenic EDCs. Furthermore, the high sensitivity of this testing method, as well as the possibility of analyzing vast datasets rapidly in a completely automated fashion indicate the huge potential for this rapid behavior test to become a sensitive, standardized, non-invasive biomarker.

Este trabalho apresenta o desenvolvimento e validação de métodos analíticos para investigar a presença de fármacos e seus metabólitos, assim como disruptores endócrinos, em amostras de águas superficiais, utilizando estratégias de \"clean up\" e enriquecimento de amostra no modo \"on line\" (\"stacking\") e \"off line\" (extração em fase sólida, SPE), em junção com técnicas de separação avançadas em meio líquido (eletroforese capilar, CE, e cromatografia a líquido, LC, com detecção UV, e seus acoplamentos com espectrometria de massas). No primeiro Capítulo são discutidos aspectos gerais sobre os produtos farmacêuticos, produtos de higiene pessoal e disruptores endócrinos, bem como a origem e ocorrência destas substâncias no meio ambiente. O segundo Capítulo aborda o desenvolvimento de um método de separação capaz de determinar oito substâncias entre fármacos de caráter ácido e metabólitos (diclofenaco, bezafibrato, fenoprofeno, ibuprofeno, cetoprofeno, naproxeno e ácidos gentísico e salicílico) numa única corrida, utilizando eletroforese capilar com enriquecimento em linha da amostra (stacking do analito baseado em grande volume de injeção da amostra) utilizando eletrólito de corrida constituído por 30 mmol L-1 de tetraborato de sódio e 5 mmol L-1 de Brij 35, pH 9,3. O método proposto alcançou limites de detecção que variaram de 2 µg L-1 para o fármaco naproxeno até 80 µg L-1 para o ibuprofeno. No terceiro capítulo é explorado um método de separação por CE para nove substâncias entre fármacos e hormônios (fluoxetina, trimetoprima, diazepam, carbamazepina, propranolol, clofibrato, fenofibrato, etinilestradiol e estrona). Utilizou-se como estratégia de pré-concentração dos analitos, o modo \"stacking\" de micelas com grande volume de injeção de amostra. Este método chegou a limites de detecção na ordem de 9 µg L-1 com eletrólito de corrida composto por 30 mmol L-1 de ácido fosfórico, 40 mmol L-1 de dodecilsulfato de sódio, 20% (v,v) de acetonitrila e 0,1% (v,v) de trietilamina. No quarto capítulo, foi realizado o estudo de parâmetros físico-químicos que estão relacionados à técnica de extração em fase sólida, tais como tipo do sorvente, volume de capacidade, volume de eluição, lavagem do cartucho de extração, entre outros. As condições ótimas de \"clean up\" e pré-concentração \"off line\" da amostra obtidas foram combinadas com as condições ótimas de pré-concentração \"on line\" e separação descritas nos Capítulos 2 e 3, para todas as substâncias abordadas ali, com o intuito de analisar amostras reais de água superficial coletadas no reservatório Billings (Estado de São Paulo). O método combinado permitiu alcançar concentrações da ordem de 500 ng L-1, com valores de recuperação satisfatórios (58 - 88%), quando levada em consideração a origem complexa da matriz ambiental. No quinto capítulo, desenvolveu-se um método de análise de p-hidroxibenzoatos de alquila, substâncias utilizadas como conservantes em diversos produtos de uso diário, utilizando eletroforese capilar associada a estratégias de concentração \"on line\" (stacking com injeção de grande volume de amostra) e \"off line\" (SPE). O método proposto, utilizando eletrólito constituído por 40 mmol L-1 de glicina e 40 mmol L-1 de trietilamina, foi aplicado na análise destas substâncias em amostras de água superficial, alcançando níveis de concentração da ordem de 4 6 µg L-1. O sexto capítulo aborda o desenvolvimento de um método por eletroforese capilar destinada à análise de alquilbenzeno sulfonato linear (LAS) e homólogos, tensoativo comumente utilizado na composição de detergentes de uso doméstico e industrial. O eletrólito de separação era composto de 60 mmol L-1 TRIS, 30 mmol L-1 HIBA, 15 mmol L-1 Brij 35 e 40% (v,v) acetonitrila. Foi realizada uma etapa de extração em fase sólida (C18), e uma concentração total de LAS na ordem de 1,09 mg L-1 foi encontrada em uma amostra de efluente de estação de tratamento de esgoto. No capítulo 7 foi desenvolvido um método utilizando eletroforese capilar acoplada a um espectrômetro de massas, com analisador \" ion trap\", para a análise dos fármacos cimetidina, propranolol, salbutamol, trimetoprima e metoclopramida. Amostras de água coletadas no reservatório Billings foi fortificada com os fármacos em estudo e submetida a procedimento de extração em fase sólida em cartuchos de poliestireno divinilbenzeno (PS-DVB). O método permitiu a análise das substâncias estudadas na concentração aproximada de 40 µg L-1. Finalmente, no oitavo capítulo foi explorado um método envolvendo cromatografia líquida de alta eficiência, hifenada a dois tipos de espectrômetros de massas: um triplo quadrupolo e um triplo quadrupolo com \"ion trap\" linear. Foram investigados fármacos de diversas classes em amostras de água coletadas no reservatório Billings, sendo possível encontrar carbamazepina na concentração de 20 ng L-1, com apenas uma etapa de filtração da amostra antecedendo a análise, utilizando o triplo quadrupolo. Cabe destacar que o LOD para este analito foi de 400 fg L-1, sem nenhum tratamento da amostra visando préconcentração. / This work presents the development and validation of analytical methods to investigate the presence of pharmaceutical compounds, their metabolites and endocrine disruptors in surface water using on line (stacking) and off line (solid phase extraction) sample clean up and enrichment strategies coupled to advanced separation techniques in liquid medium (capillary electrophoresis and liquid chromatography with UV detection and their hyphenation with mass spectrometry). In the first Chapter, general aspects on pharmaceuticals, products of personal care and endocrine disruptors are discussed as well as their origin and means of entry to the environment. Chapter 2 describes the development of a separation method for the determination of eight pharmaceuticals and endocrine disruptors substances with acidic character (diclofenac, bezafibrate, fenoprofen, ibuprofen, ketoprofen, naproxen, gentisic and salicylic acids) in a single run using capillary electrophoresis with on line sample enrichment (analyte stacking with large sample volume injection) in electrolytes composed of 30 mmol L-1 sodium tetraborate at pH 9.3 and 5 mmol L-1 Brij 35. The proposed method reached limits of detection between 2 µg L-1 (naproxen) and 80 µg mL-1 (ibuprofen). In Chapter 3, a CE separation method for the determination of nine pharmaceuticals and hormones with neutral and basic character (fluoxetin, trimethoprim, diazepam, carbamazepine, propranolol, clofibrate, fenofibrate, ethynylestradiol and estrone) was exploited. As preconcentration strategy, micelle stacking with large sample volume injection was performed. Limits of detection in the order of 9 µg L-1 were reached with electrolytes composed of 30 mmol L-1 phosphoric acid, 40 mmol L-1 sodium dodecylsulfate, 20% (v,v) acetonitrile and 0.1% (v,v) triethylamine. In Chapter 4, the physicochemical parameters associated with the solid phase extraction technique, such as sorbent type, breakthrough volume, elution volume, extraction cartridge rinse, among others, were evaluated. Optimum sample clean up and off line preconcentration conditions combined with the optimum on line preconcentration and separation conditions described in Chapters 2 and 3, for all substances under consideration, were applied to the analysis of real surface water samples collected at the Reservoir Billings (Sao Paulo state, Brazil). The combined method reached concentrations in the order of 500 ng L-1, with satisfactory recoveries (58 88%) for complex environmental matrices. In Chapter 5, a method for the analysis of alkyl p-hydroxybenzoates, substances used as preservatives in several products of personal care, was developed using capillary electrophoresis associated with on line (stacking with large volume injection) and off line (SPE) preconcentration strategies. The proposed method, which used 40 mmol L-1 glycine and 40 mmol L-1 triethylamine as electrolyte, was applied to the analysis of alkyl p-hydroxybenzoates in surface water, reaching 4 - 6 µg L-1 concentrations. Chapter 6 describes the development of a CE method for de analysis of linear alkylbenzene sulfonates (LAS) and homologues, surfactants commonly used in the composition of detergents of industrial and domestic use. As separation electrolyte, 60 mmol L-1 TRIS, 30 mmol L-1 HIBA, 15 mmol L-1 Brij 35 and 40% (v,v) acetonitrile was used. A solid phase preconcentration extraction step in C18 was employed and a total LAS concentration of 1.09 mg L-1 was found in a sample obtained from the effluent of a sewage treatment plant. In Chapter 7, a CE-MS (ion trap) method was developed for the analysis of cimetidine, propranolol, salbutamol, trimethoprim and methoclopramide in fortified surface water samples collected in the Reservoir Billings (Sao Paulo state, Brazil) and previously enriched by SPE (PS-DVB). The method reached concentrations of c.a. 40 µg L-1. Finally, in Chapter 8, a method based on high-performance liquid chromatography coupled with two different mass spectrometers: a triple quadrupole and a triple quadrupole with linear ion trap). Several pharmaceuticals were investigated in surface water samples collected from the Reservoir Billings (Sao Paulo state, Brazil). With the LC-MS/MS (triple quadrupole) system, carbamazepine was found in a non treated sample (just a filtration step prior to injection was performed) in a concentration level of 20 ng L-1. It is worth mentioning that for carbamazepine, a LOD of 400 fg L-1 was found, without any preconcentration sample treatment

L’objectif de ce projet a été de développer une approche bio-analytique permettant l’évaluation du danger inhérent à la multi-contamination de l’air intérieur en Ile-de-France. Des méthodes d’analyse chromatographique couplée à la spectrométrie de masse ont été développées et validées pour 62 composés d’intérêt présentant un potentiel perturbateur endocrinien (PE) avéré ou suspecté. Le potentiel PE a été évalué sur des bio-essais cellulaires de mesure de perturbation d’activité transcriptionnelle. Les résultats montrent que les familles de composés majoritaires dans l’air intérieur sont, par ordre décroissant : phtalates > muscs synthétiques > alkylphénols > parabènes. En outre, les composés sont prédominants en phase gazeuse, et les habitats les plus contaminés sont la crèche et la maison. L’air intérieur présente un potentiel PE de type œstrogénique, thyroïdien et anti-androgénique. En accord avec son profil de contamination, l’activité biologique de ce dernier se concentre majoritairement dans la phase gazeuse, et tend à être plus élevée dans la crèche et la maison. Une analyse dirigée par les bio-essais, ou effect-directed analysis (EDA), a été mise en œuvre pour identifier les composés cibles à l’origine des effets PE de l’air intérieur. Les composés suivants ont été identifiés comme étant potentiellement à l’origine d’effets PE observés : les phtalates, le méthyl-parabène, les alkylphénols, la cyperméthrine et les muscs synthétiques. Ce travail apporte des connaissances sur le danger inhérent à la multi-contamination de l’air intérieur ainsi que des données d’exposition utiles à une évaluation des risques sanitaires. / The objective of this project was to develop a bio-analytical approach leading to the assessment of the inherent hazard of the indoor air multi-contamination. Chromatographic methods combined with mass spectrometry were developed and validated for 62 target molecules known or suspected as endocrine-disrupting (ED) compounds. The ED potential was assessed by cellular bioassays measuring perturbations of transcriptional activity. The data showed that the predominant families of compounds in indoor air were in the following descendant order: phthalates > musks > alkylphenols > parabens. The ED contaminants were mainly present in gaseous phase, and the most contaminated locations were the day nursery and the house. An estrogenic, thyroid and anti-androgenic potential was attributed to indoor air. In agreement with its contamination profile, the biological activity of the latter was concentrated predominantly in the gaseous phase, and tended to be higher in the day nursery and the house. An effect-directed analysis (EDA) was carried out to identify the target chemicals responsible for the ED effects of indoor air. The following chemicals were identified as being potentially responsible for the observed ED effects: phthalates, methyl-paraben, alkylphenols, cypermethrin and synthetic musks. This work provides both knowledge about the inherent hazard of the indoor air multi-contamination and exposure data useful in health risk assessment.

Orientador: Wellerson Rodrigo Scarano / Resumo: Citocinas e proteínas quinases são fundamentais para o controle do processo espermatogênico, estando diretamente envolvidas na dinâmica da barreira hematotesticular. Diferentes mecanismos de controle são modulados por receptores como o GPR30, que ativa rapidamente diferentes vias de sinalização, responsáveis pelos processos de proliferação, sobrevivência e morte celular. Os desreguladores endócrinos (DEs) possuem grande afinidade pelo GPR30, além de potencial para ativar vias de estresse oxidativo e a abertura da barreira. Antagonistas funcionais dos DEs, como o Panax ginseng, podem ser protetores contra seus efeitos. Considerando a importância das vias de sinalização que regulam a espermatogênese e a constante exposição ambiental aos DEs a que estamos submetidos, este trabalho objetiva estudar a possível modulação da via não genômica ativada por GPR30 e do estresse oxidativo em células de Sertoli expostas a baixas doses do DE Monobutil Ftalato (MBP) bem como o potencial papel citoprotetor do GIM-1 (metabólito do P. ginseng) sobre essas células. Para tal, as células de Sertoli humanas (HSec) foram mantidas sobre matriz artificial, simulando o ambiente in vivo. A exposição ao MBP e ao GIM-1 foi realizada nos tempos de 30min, 1, 12, e 48 horas, em doses pré-estabelecidas pelo ensaio do MTT (teste de toxicidade) em 4 grupos: controle, MBP, GIM-1 e MBP + GIM-1. A morfologia celular foi avaliada pela coloração com Hematoxilina e Eosina, evidenciando efeitos deletérios do MBP sobre... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Cytokines and kinases protein are essential to control the spermatogenic process, being directly involved in the blood-testis barrier control. Activation of these mechanisms is modulated by receptors such as GPR30, which rapidly activates different signaling pathways responsible for proliferation and cell death processes. Endocrine Disruptors (EDs) have high affinity for GPR30, causing oxidative stress and possible barrier rupture. Functional antagonists of EDs, such as Panax ginseng, may be protective against their effects. Considering the importance of the signaling pathways that regulate spermatogenesis and the constant environmental exposure to the EDs to which we are subject, this work aims to study the possible modulation of the non-genomic pathway activated by GPR30 and oxidative stress in Sertoli cells exposed to low doses of the ED Monobutyl Phthalate (MBP) and the possible cytoprotective role of GIM-1 (P. ginseng metabolite) on these pathways. To this end, HSec human lineage cells were maintained on artificial matrix, simulating in vivo environment. Exposure to MBP and GIM-1 was performed at 30 min, 1, 12 and 48 hours at pre-set MTT (toxicity assay) levels in 4 groups: control, MBP, GIM-1 and MBP + GIM-1. Morphology and cell adhesion were evaluated by staining with Hematoxylin and Eosin, evidencing deleterious effects of MBP above cell distribution and adhesion in basement membrane; GIM-1 group was similar to Control and MBP+GIM-1 showed an intermediate aspect. In o... (Complete abstract click electronic access below) / Doutor

De nombreuses affections chez l’enfant et l’adulte sont associées à une exposition in utero à des perturbateurs endocriniens (PE). Pour réduire cette exposition, des programmes d’éducation pour la santé environnementale périnatale se développent. Ils font évoluer des dimensions psychosociales que sont la perception du risque (PR) et la croyance en l’action de réduction de l’exposition (CAR), mais peu d’entre eux sont évaluésL’étude de recherche interventionnelle PREVED a pour objectif d’évaluer l’efficacité d’un programme d’éducation pour la santé environnementale pour des femmes enceintes, visant à diminuer leur exposition aux PE.Les objectifs de ces travaux métrologiques étaient de développer des outils (i) analytiques et (ii) épidémiologiques en vue d’évaluer l’efficacité du programme. Ainsi, (i) des méthodes analytiques ultrasensibles par LC-MS/MS permettant de doser les fractions non conjuguées de PE dans les urines et le colostrum ont été développées et validés sur des prélèvements recueillis dans le cadre de la cohorte périnatale EDDS; (ii) un questionnaire psychosocial explorant la PR, la CAR et les connaissances des femmes enceintes au moyen de scores a été conçu.Ces travaux s’inscrivent dans une approche interdisciplinaire de santé environnementale. Ils proposent des méthodes analytiques fiables pour mesurer l’exposition aux PE étudiés d’une part, et un questionnaire évaluant connaissances, attitudes et pratiques des femmes enceintes sur la question des PE d’autre part. Ces outils permettront de mesurer l’impact du programme d’éducation pour la santé environnementale périnatale. / Childhood and adulthood diseases are associated to in utero exposure to endocrine disrupting chemicals (EDCs). To reduce this exposure, environmental health education programs dedicated to perinatal period are implemented. These programs change psychosocial dimensions such as risk perception (RP) and belief in the action of exposure reduction (BAR), but few are assessed.The interventional research PREVED study aims to assess efficacy of a perinatal environmental health education program to reduce pregnant women exposure to EDC.The objectives of this work were to develop analytical (i) and (ii) epidemiological tools to evaluate the efficacy of the program. Thus, (i) ultrasensitive LC-MS/MS analytical methods were developed to determine unconjugated fractions of EDCs in urine and colostrum and validated using samples collected from the EDDS perinatal cohort; (ii) a psychosocial questionnaire exploring RP, BAR and knowledge of pregnant women by means of scores was developed.This work, part of an interdisciplinary approach to environmental health, proposes reliable analytical methods to assess exposure to the studied EDCs on the one hand, and a questionnaire assessing knowledge, attitudes and practices of pregnant women about EDCs on the other hand. The impact of the perinatal environmental health education program will be assessed using these tools.

O alumínio (Al) é o metal mais abundante na natureza e tornou-se importante poluente aquático trazendo prejuízos a reprodução de teleósteos, atuando como um desregulador endócrino. No entanto, em experimentos in vivo não é possível demonstrar que os efeitos do Al no eixo endócrino reprodutivo são devido a sua atuação direta sobre os órgãos que o compõem. Por isso, este trabalho teve como objetivo avaliar o efeito direto do Al sobre células foliculares ovarianas, gonadotrópicas e somatolactínicas hipofisárias de fêmeas maduras de Oreochromis niloticus. Para isso dois experimentos in vitro de exposição ao Al foram realizados: um utilizando-se ovários maduros e outro hipófises de fêmeas sexualmente maduras. Para os experimentos ovarianos, frações de ovários maduros foram incubadas por 4 horas formando os seguintes grupos: 1) Grupo controle (Ctr): tecido ovariano exposto somente à solução de Krebs-Ringer-glucose-HEPES; 2) Grupo gonadotropina coriônica humana (hCG): tecido exposto à 6 µg/ml de hCG (Ovidrel); 3) Grupo (hCG+Al): tecido exposto à 6 µg/ml de hCG (Ovidrel) + cloreto de alumínio (AlCl3) - 10mM; 4) Grupo (Al): exposto somente ao AlCl3 - 10mM. A concentração dos hormônios 17β-estradiol (E2) e 17α-hidroxiprogesterona (17αOHP) no meio de incubação foi determinada por ELISA. Para os experimentos hipofisários, hipófises de fêmeas sexualmente maduras foram incubadas por 24 horas formando os seguintes grupos: 1) Ctr: hipófise exposta somente ao meio L15 (controle interno); 2) GnRH: hipófise exposta somente ao GnRH (controle de liberação de gonadotropinas); 3) GnRH+Al: hipófise exposta ao GnRH + AlCl3 10mM e 4) Al: hipófise exposta somente AlCl3 10mM. Após o período experimental, foram realizadas análises de qPCR (PCR quantitativo), análises de imunohistoquímica, e de microscopia eletrônica. Os resultados do experimento ovariano mostraram que os fragmentos ovarianos do grupo exposto à hCG apresentaram um aumento significativo da liberação dos hormônios E2 e 17αOHP em relação aos demais grupos, confirmando o efeito desta gonadotropina sintética na liberação destes esteroides gonadais. No entanto, a administração conjunta da hCG com Al (hCG+Al) não gerou este aumento da produção dos esteroides em relação ao grupo controle. Estes dados evidenciam que o Al inibiu a resposta celular das células esteroidogênicas ovarianas às gonadotropinas. Os resultados dos experimentos hipofisários mostraram que o Al (GnRH+Al e Al) afetou a expressão gênica dos genes estudados (βFSH, (βLH, SL) inclusive dos normalizadores (EF1α e (βAc), o que tem sido comum em experimentos de ecotoxicologia. Os dados de microscopia eletrônica mostraram desestruturação celular nas hipófises que foram expostas ao Al e as análises de imunohistoquímica apontaram que o Al (GnRH+Al e Al) não interferiu sobre a quantidade de grânulos de βLH e SL, enquanto o grupo Al indicou uma diminuição na quantidade de grânulos de βFSH, sugerindo que o Al afeta a dinâmica de síntese/liberação desta gonadotropina. Estes dados evidenciam a toxicidade do Al diretamente sobre ambos os órgãos estudados, tanto em nível de resposta celular quanto em nível estrutural confirmando o potencial do Al como um DE e amplia as perspectivas de estudo sobre o mecanismo de ação do Al como um DE / Aluminum (Al) is the most abundant metal in nature and has become an important water pollutant impairing reproduction of teleosts, acting as an endocrine disrupter. However, in vivo experiments cannot demonstrate that the effects of Al on the reproductive endocrine axis are due to direct action on the organs that compose it. Therefore, this study aimed to assess the direct effect of Al on ovarian follicular cells, gonadotropic and somatolactin pituitary cells of mature females of O. niloticus. For this, two in vitro exposure experiments of Al were performed: one using mature ovaries and other using pituitaries of sexually mature females. For ovarian experiments, fractions of mature ovaries were incubated for 4 hours to obtain the following groups: 1) control group (Ctr): ovarian tissue only exposed to Krebs- Ringer-HEPES-glucose; 2) human chorionic gonadotropin (hCG) group: tissue exposed to 6 mg/ml hCG (Ovidrel); 3) hCG + Al group: tissue exposed to 6 mg/ml hCG (Ovidrel) + aluminum chloride (AlCl3) - 10mM; 4) Al group: only exposed to 10 mM AlCl3. The concentration of the hormones 17β-estradiol (E2) and 17α-hydroxyprogesterone (17αOHP) in the incubation medium was determined by ELISA. For pituitary experiments, pituitaries of sexually mature females were incubated for 24 hours to form the following groups: 1) Ctr: pituitary exposed only to L15 (internal control); 2) GnRH: only exposed to the pituitary GnRH (gonadotropin releasing hormone); 3) GnRH + Al: exposed to the pituitary GnRH AlCl3 + 10 mM and 4) Al: 10mM AlCl3 only exposed pituitary. After the assay period, analysis of qPCR (quantitative PCR), analysis of immunohistochemistry and electron microscopy were performed. The results of the experiment showed that the ovarian exposed to hCG group showed a significant increase in the release of E2 and 17αOHP compared to the other groups, confirming the effect of synthetic gonadotropin in the release of these gonadal steroids. However, the administration combined of hCG with Al (Al + hCG) did not generate this increased production of steroids compared with the control group. These data show that Al inhibited the cellular response of the ovarian steroidogenic cells to gonadotropins. The results of the experiments with pituitaries showed that Al (GnRH + Al and Al) affected the gene expression of the genes studied (βFSH, (βLH, SL) including the house keeping genes (EF1α and βAc), what has been common in ecotoxicology experiments. Data from electron microscopy showed cell disruption in the pituitary glands that were exposed to Al and immunohistochemical analyzes showed that Al (GnRH + Al and Al) did not affect the amount of granules of βLH and SL, while the Al group indicated a decrease the amount of βFSH granules, suggesting that Al affects the dynamics of the synthesis/release of this gonadotropin. These data show the toxicity of Al directly on both organs studied, at both the cellular response as for structural level confirming the potential of Al as a DE and increases the perspectives of study on the mechanism of action of Al as a DE