No description available.

Fluoxetine, a selective serotonin re-uptake inhibitor (SSRI) and the active ingredient in Prozac®, is found in the environment and disrupts feeding and metabolism in exposed fish. The objective of this research was to investigate the mechanisms involved in the feeding and metabolism disruption in the model goldfish (Carassius auratus). Two short-term waterborne fluoxetine exposures (7- and 14-days) were performed using two environmentally relevant doses of fluoxetine (0.5 and 1 μg/L) and metabolic effects at the level of the brain, liver, serum and bile in goldfish were investigated. Abundances of mRNA transcripts coding for six feeding neuropeptides were examined to determine which may be involved in the initial neural changes associated with decreased appetite in goldfish. The 7-day fluoxetine exposure at 1 μg/L caused corticotropin-releasing factor (CRF) mRNA levels to increase by 2-fold in female hypothalamus and telencephalon, indicating that CRF may be one of the first of the feeding neuropeptides to be altered. Six hepatic miRNAs were also evaluated in the goldfish liver that were previously associated with fluoxetine exposure in zebrafish (Danio rerio). Following the 7-day exposure at 1 μg/L, miR-22b, miR-140, miR-210, miR-301a and miR-457b levels increased in the female goldfish liver by 4-6 fold. The 14-day fluoxetine exposure at 1 μg/L caused 2-fold increases in miR-210, miR-301a, miR-457b and let-7d in male goldfish liver. These miRNAs were associated with the down-regulation of anabolic metabolic pathways in zebrafish, indicating a conservation of miRNA and fluoxetine effect between fish species. Serum and bile metabolite profiles of fluoxetine exposed goldfish were evaluated using ultra performance liquid chromatography coupled to quadrupole time of flight mass spectrometry. Following the 14-day exposure at 1 μg/L, the bile metabolite profiles of male goldfish were significantly different from controls as detected by cluster analysis and fluoxetine was tentatively identified in the serum. No other discriminant metabolites were identified as of yet. The data presented suggest that fluoxetine causes metabolic disruption in goldfish at multiple organ levels. Because of the widespread detection of fluoxetine and other emerging SSRIs in the aquatic environment, future research is required to firmly establish this pharmaceutical class as a metabolic and endocrine disrupting chemical.

Developing a sensitive and effective in vitro bioassay to detect endocrine disrupting chemicals (EDCs) would reduce the cost, eliminate the possibility of low dose effects, detect the non-monotonic dose responses, and identify mechanisms of actions. The “chemical nose” sensing method using supramolecular complexes composed of cationic monolayer functionalized gold nanoparticles (AuNPs) and fluorescent proteins (FPs) can successfully distinguish serum proteins, mammalian cells, tissue lysates, and chemotherapeutic drug mechanisms. EDCs regulate target cells via genomic or non-genomic pathways in terms of proliferative effect and response time. In this thesis, green fluorescent protein-gold nanoparticle (GFP-AuNP) sensors were used to detect the proliferative effect of 17b-estradiol (E2) and bisphenol A (BPA) on MCF7 and T47D cell lines at fM or pM dose range. Non-monotonic dose responses were also observed at different exposure times. The dose-response relationships using GFP-AuNP sensors could be correlated to the cell cycle analysis. Interestingly, tamoxifen, an estrogen antagonist, showed distinct patterns at low doses on HepG2 cells using triple channel FP-AuNP sensors, which might indicate different mechanisms of actions in this dose range.

Endocrine disrupting chemicals (EDCs) are ubiquitous in the environment and have been detected in drinking water. Although various water treatment processes can remove EDCs, chemicals can migrate from pipes that transport water and contaminate drinking water. Globally bottled water consumption is steadily rising as an alternative to tap water, but EDCs have also been detected in bottled water. Sources of EDCs in bottled water include contamination of the water source, contamination through the production process or the migration of EDCs from the packaging material. There is limited information on EDCs in drinking water and bottled water from South Africa. The aim of this study was to determine the estrogenic activity, levels of selected EDCs and the potential health risks associated with the consumption of water from selected distribution points in Pretoria (City of Tshwane) and Cape Town as well as bottled water. The study consisted of 3 phases. Phase 1 included the analysis of drinking water samples from ten water distribution points in Pretoria and Cape Town collected over four sampling periods. In phase 2, ten brands of bottled water were analysed after exposure to different storage conditions (20°C, 40°C, light and dark) for 10 days. Samples were extracted using a C18 solid phase extraction method. Estrogenic activity was assessed using the recombinant yeast estrogen screen (YES) bioassay and the T47D-KBluc reporter gene bioassay. The extracts were analysed for di(2-ethylhexyl) adipate (DEHA), di(2-ethylhexyl) phthalate (DEHP), diisononylphthalate (DINP), dibutyl phthalate (DBP), bisphenol A (BPA), nonylphenol (NP), 17β-estradiol (E2), estrone (E1) and ethynylestradiol (EE2) using UPLC-MS. Phase 3 consisted of a scenario based health risk assessment to assess the carcinogenic and toxic human health risks associated with the consumption of distribution point and bottled water. All the samples were below the detection limit (dl) in the YES bioassay, but estrogenic activity was detected in bottled and distribution point water using the T47D-KBluc bioassay. All samples were below the 0.7 ng/L trigger value for estrogenic activity in drinking water. NP was below the dl for all the samples, E2 was detected in five distribution point samples and E1, EE2, DEHA, DEHP, DINP, DBP and BPA were detected in distribution point and bottled water samples. The estrogenic activity and levels of target chemicals were comparable to the levels found in other countries. Hazard quotients for BPA, DEHA and DINP were higher in bottled water compared to distribution point water. The greatest non-carcinogenic health risk was posed by E1 in distribution point water from Pretoria and the highest cancer risk by levels of DEHP in distribution point water from Cape Town. However, overall, health risk assessment revealed acceptable health and carcinogenic risks associated with the consumption of distribution point and bottled water. Although the potential health risks posed by the EDCs found in the water samples in this study were low, the fact that potential EDCs were found in the water samples are still of concern. A monitoring strategy that also includes water from other municipalities and other brands of bottled water are therefore recommended. / Thesis (PhD)--University of Pretoria, 2016. / School of Health Systems and Public Health (SHSPH) / PhD / Unrestricted

Orientador: Luis Fernando Barbisan / Coorientador: Wellerson Rodrigo Scarano / Banca: Daniel Araki Ribeiro / Banca: Luis Antonio Justulin Junior / Resumo: O crescimento, a maturação e a manutenção estrutural e funcional da próstata são eventos hormônio-dependentes, principalmente de andrógenos, e alterações endócrinas no período crítico de desenvolvimento prostático poderão causar efeitos adversos sobre a próstata, inclusive câncer. Bisfenol A (BPA) é um desregulador endócrino, presente em embalagens de alimentos e detectado em praticamente todos os fluidos humanos, capaz de alterar o desenvolvimento do trato reprodutivo de ratos machos por meio de exposição gestacional/pré-natal. A genisteína (GEN), um fitoestrógeno da soja, mostrou propriedades quimiopreventivas contra carcinogênese em roedores. Este estudo objetivou avaliar se a exposição gestacional ao BPA causa efeitos deletérios precoces e tardios sobre a morfogênese prostática da prole de machos Sprague-Dawley, e se a exposição concomitante à GEN modificaria estes efeitos deletérios. Fêmeas prenhes foram divididas em grupos que receberam BPA (25 ou 250 μg/kg p.c.) ou dimetilsulfóxido (DMSO - grupo controle)/óleo de canola, por gavage e dieta basal ou contendo GEN (250 μg/kg). A prole recebeu apenas ração basal e foi eutanasiada nos dias pós-natal (DPN) 21 ou 180. A exposição gestacional à menor dose de BPA induziu alterações prostáticas precoces e tardias. A ingestão gestacional de GEN reverteu os efeitos deletérios do BPA sobre os níveis de proliferação celular no epitélio prostático, a arquitetura prostática e a expressão de receptores androgênicos na prole no DPN 21. Um aumento na incidência de inflamação multifocal e de hiperplasia prostática atípica foi observado na prole, no DPN 180, das mães tratadas com a menor dose de BPA, enquanto a ingestão gestacional de GEN atenuou estes efeitos. Desta forma, os resultados do presente estudo indicam que a ingestão de GEN durante a gestação tem ação preventiva contra efeitos adversos do BPA sobre a próstata da prole / Abstract: Not available / Mestre

Notre étude a porté sur le diagnostic de la présence des perturbateurs endocriniens dans le golfe de Gabès (Tunisie), leurs effets sur la reproduction d’une espèce bioindicatrice de pollution, la palourde R. decussatus ainsi que leurs mécanismes d’action. Pour atteindre ces objectifs, nous avons fait appel à plusieurs outils méthodologiques, des expositions in vivo (effluents urbains, effluents industriels, 17β-œstradiol) et des prélèvements in situ afin d’évaluer le risque encouru par l’écosystème du golfe de Gabès tout entier en utilisant une approche multiparamétrique basée sur différents niveaux de l’organisation biologique (allant du niveau biochimique jusqu’au niveau populationnel). Nous avons mis en évidence la présence des composés reprotoxiques tout au long de la côte Sud-Est de la Tunisie. Les résultats obtenus ont révélé une corrélation entre la contamination chimique (cadmium, activités œstrogéniques et androgéniques au niveau du sédiment) et certains paramètres biologiques liés à la reproduction (diamètre ovocytaire, vitellogénine, glycogène) et un site très divergent de tous les autres sites (El hofra). Cette station d’El hofra semble être touchée par une perturbation endocrinienne liée probablement à une synergie entre la contamination par le Cd et la présence des composés à activité œstrogénique et androgénique dans le milieu. En parallèle à cette étude in situ, trois expérimentations au laboratoire ont été réalisées. Les résultats ont montré que les effluents de la station d’épuration de Sfax (cocktail de contaminants) sont capables d’affecter la reproduction de la palourde en induisant la synthèse de la vitellogénine et stimulant la reproduction chez les deux sexes. L’exposition des palourdes à un contaminant pur, le 17β-œstradiol, a révélé qu’il peut contribuer à expliquer les perturbations du cycle reproducteur observées dans l’expérience précédente. Cette hormone naturelle a un effet et sans doute un rôle dans la physiologie de la palourde. Dans le but d’étudier son mécanisme d’action via le récepteur aux œstrogènes (RE), nous avons cloné une partie de l’ADNc codant pour les RE. Cependant, l’analyse de la réponse transcriptionnelle n’a pas montré de changement significatif dans l'expression du gène aussi bien chez les mâles que chez les femelles suite à une exposition aux œstrogènes (17β-œstradiol, effluents urbains et industriels). / Our study aimed to assess the presence of endocrine disrupting compounds in the Gulf of Gabes area (Tunisia), their effects on the reproductive potential of the marine bivalve, Ruditapes decussatus as well as elucidating their mechanisms of action. To achieve these goals, several methodologies have been deployed, such as laboratory in vivo exposures (to urban or industrial effluents, estradiol), and in situ sampling to assess the risk of such substances on this ecosystem by using a multiparametric approach based on different levels of biological organization in clams (from biochemical level to the population level). We have demonstrated the presence of reprotoxic compounds along the south-eastern coast of Tunisia. The results revealed, a strong correlation between chemical contamination (cadmium, estrogenic and androgenic activities in sediment) and some reproductive parameters (oocyte diameter, vitellogénine and glycogen levels) and, a particular site (El hofra) that differs from all the other studied sites in this polluted context. Clams from this station seem to suffer from endocrine disruption probably resulting from a synergistic effect between both cadmium contamination and the presence of chemicals acting as estrogens and androgens in the environment. In parallel to this in situ study, three laboratory experiments were performed. The results showed that exposure to a mixture of chemical compounds released by the Sfax wastewater treatment plant had an adverse impact on the reproductive potential of R. decussatus, by inducing vitellogenin synthesis and stimulating reproduction in both sexes. Results of the exposure of organisms to 17β-estradiol alone, revealed that the estrogenic compound could contribute to the observed effects on the reproductive cycle of the test animals in the previous experiment. This natural hormone has induced deleterious effects and may play a role in the physiology of the clam. To study its mechanism of action via estrogen receptors, a partial cDNA of the gene was isolated and characterized from the clam R. decussatus. However, analysis of the transcriptional response did not show a significant change in the gene expression in males as well as in females following exposure to estrogens (17β-estradiol, urban and industrial effluents).

京都大学 / 新制・論文博士 / 博士(工学) / 乙第13495号 / 論工博第4202号 / 新制||工||1786(附属図書館) / (主査)教授 米田 稔, 教授 高野 裕久, 教授 松井 康人 / 学位規則第4条第2項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

No description available.

Endocrine disrupting chemicals (EDCs) are compounds that can interfere with hormone signaling pathways and are now recognized as pervasive in estuarine and marine waters. One prevalent EDC in California’s coastal waters is the xenoestrogen 4-nonylphenol (4-NP), which has been shown to impair reproduction, development, growth, and in some cases immune function of marine invertebrates. To further investigate effects of 4-NP on marine invertebrate immune function we measured total hemocyte counts (THC), relative transcript abundance of immune-relevant genes, and lysozyme activity in Pacific oysters (Crassostrea gigas) following bacterial infection. To quantify these effects we exposed oysters to dissolved phase 4-NP at high (100 μg l-1), low (2 μg l-1), or control (100 μl ethanol) concentrations for 7 days, and then experimentally infected (via injection into the adductor muscle) the oysters with the marine bacterium Vibrio campbellii. 4-NP significantly altered the effects of bacterial infection had on THC. Oysters exposed to both high and low 4-NP did not experience a bacteria-induced increase in THC, as seen in control oysters. We also determined that V. campbellii infection induced differential expression of a subset of immune-related genes tested (Cg-bigdef2, Cg-bpi1, Cg-lys1, Cg-timp) in some, but not all, tissues; 4-NP exposure altered expression patterns in two of these genes (Cg-bpi1 and Cg-tgase). Exposure to 4-NP alone also caused differential expression in some genes (Cg-bpi1, Cg-galectin1, Cg-clec2). Lastly, low levels of 4-NP significantly increased lysozyme activity 24 h post-infection. These results suggest that exposure to 4-NP can alter both cellular and humoral immune responses to bacterial infection in C. gigas.

The full text will be available at the end of the embargo period: 28th March 2027