In two separate experiments, weight adjusted doses of nocodazole and acrylamide were injected intraperitoneally at various time intervals into twelve week old female mice. Within the nocodazole experiment, the doses were injected at varying time intervals before and after mating. On day seventeen of gestation, the female mice were sacrificed and their uterine contents examined. Nocodazole induced a significant increase in reproductive pathology per total implants when administered one hour after mating to the (SECxC57BL)F, stock: 5.00% total deads, 70.23% moles, and 3.41% abnormal fetuses. Acrylamide treatment produced a significant reduction in live births when administered six hours after mating: 50.86% moles and 46.46% living fetuses per total implants.

Dans une première partie du travail, nous nous sommes intéressés à l'effet de l'acrylamide sur la tumorigénèse thyroïdienne. En effet des chercheurs suédois avaient récemment attiré l’attention de la communauté scientifique sur la présence de ce xénobiotique dans des denrées alimentaires couramment utilisées. Des études antérieures avaient montré que l’acrylamide, administré chroniquement à des rats pendant deux ans, induisait des tumeurs parmi lesquelles les tumeurs thyroïdiennes étaient les plus fréquentes. Pour ces raisons l’acrylamide est considéré comme une substance potentiellement cancérigène pour l’homme. Nous avons voulu investiguer les mécanismes d’action possibles de l’acrylamide dans la tumorigénèse thyroïdienne avec un modèle in vitro et ensuite un modèle in vivo.<p>Dans le modèle in vitro, nous avons d’abord testé l’effet de l’acrylamide sur les propriétés caractéristiques de la thyroïde, qui pourraient expliquer la spécificité de cette substance pour ce tissu. Dans un premier temps, nous avons donc testé l’effet de l’acrylamide sur l’accumulation d’AMPc (médiateur de la croissance) et sur la génération d’H2O2 potentiellement cancérigène mais nous avons montré qu’aucun de ces deux paramètres n’est modulé suite au traitement des cellules thyroïdiennes. Nous avons ensuite montré que l’acrylamide n’avait pas d’effet sur la prolifération dans les lignées thyroïdienne de rat, un facteur qui aurait pu expliquer l’effet tumorigène de l’acrylamide mais pas sa spécificité thyroïdienne. Nous avons enfin testé en utilisant l’essai comète la capacité de l’acrylamide à générer des cassures au niveau de l’ADN. Ceci nous a permis de montrer que l’acrylamide était capable d’induire des cassures dans l’ADN de lignées thyroïdiennes de rat ainsi que dans des cultures primaires de thyroïde humaines, de chien et de mouton. Afin de déterminer que l’effet observé sur l’ADN était dû à des cassures double brin, nous avons utilisé une technique de détection de l’histone H2AX phosphorylée. En effet, la phosphorylation de cette histone se produit lorsque des cassures d’ADN double brin sont présentes. Les résultats obtenus ne soutiennent pas l’hypothèse que l’acrylamide provoque des cassures double brin. Néanmoins ces dommages à l’ADN sont susceptibles d’induire des mutations qui peuvent jouer un rôle dans le processus de tumorigénèse de l’acrylamide.<p>Nous avons donc testé l’effet de l’acrylamide sur un modèle in vivo. Pour cela nous avons traité des souris avec de l’acrylamide additionné à l’eau de boisson à des doses comparables à celles administrées aux rats pendant 2, 6 ou 8 mois. Ce traitement a également été combiné avec de la thyroxine (T4) afin de mettre la thyroïde au repos ou avec du méthimazole qui inhibe la sécrétion des hormones thyroïdiennes et par conséquent induit la sécrétion de TSH, ce qui a pour effet de stimuler la glande. Ces traitements modérés ont eu les effets attendus au niveau des taux de TSH et de T4 circulants ainsi que sur la morphologie de la glande. L’acrylamide a eu de faibles effets sur le système nerveux périphérique traduit par une paralysie des membres postérieurs. Par contre nous n’avons pas observé d’apparition de tumeurs dans la thyroïde des souris. La cible thyroïdienne de l’acrylamide chez le rat semble donc spécifique de l’espèce et jette un doute sérieux sur un rôle cancérigène potentiel chez l’homme au niveau thyroïdien.<p><p>Dans une deuxième partie, nous avons investigué la réexpression de certains gènes thyroïdiens dans des lignées cancéreuses humaines. En effet nous avons montré au sein du laboratoire que la plupart des lignées thyroïdiennes les plus utilisées avaient perdu les gènes de différentiation spécifiques de la thyroïde. Pour cela nous avons utilisé des agents capables de modifier soit la méthylation de l’ADN comme la 5-aza-2’-déoxycytidine (5-AzadC) soit la compaction de la chromatine comme la trichostatine A (TSA), ce qui a pour conséquence de moduler le niveau d’expression des gènes. Ces traitements ont été utilisés à différentes concentrations, différents temps, seuls ou en combinaison. La 5-AzadC, utilisée seule a permis de réexprimer fortement les gènes Duox1 et Duox2 et faiblement NIS. En combinaison avec la TSA et la forskoline (un activateur de la voie AMPc), nous avons montré une forte réexpression de NIS mais au stade actuel de notre étude celui-ci n’est pas fonctionnel. Ceci montre que les agents modifiant la chromatine peuvent influencer l’expression des gènes de différenciation, cependant nous devons investiguer de façon plus large les différents agents ainsi que les conditions de culture pouvant mener à la réexpression d’un NIS fonctionnel. Grâce à ce traitement différenciant, qui permettrait un rétablissement du transport de l’iodure, un traitement par l’I131 des cancers indifférenciés de la thyroïde pourrait être réenvisagé.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

Orientador: Samuel Monroe Cohen / Resumo: O aumento da incidência de distúrbios testiculares e a possível influência de substâncias químicas ambientais, como o dibutilftalato (DBP) e a acrilamida (AA), exigem a identificação de modos de ação. A maioria dos estudos de toxicologia reprodutiva utiliza amostras de RNA provenientes de todo o testículo para avaliar a expressão genica; entretanto, análises de tipos celulares isolados poderiam gerar resultados mais específicos. Entre as células germinativas testiculares, as espermatogônias são importantes pois representam o início da espermatogênese. Este estudo objetivou, 1) estabelecer técnica de isolamento de espermatogônias; 2) aplicar esta técnica para verificar possíveis alterações na expressão gênica (Pou5f1, Kitlg, Mki-67, Bak1 e Spry4) em testículos de ratos pré-púberes (DPN24) e púberes (DPN45) após exposição in utero e pós-natal ao DBP ou à AA. A técnica foi eficiente para o isolamento das espermatogônias. A exposição ao DBP levou à redução do peso corporal da ninhada ao nascer, da distância anogenital dos filhotes machos no DPN4 e ao aumento da frequência de retenção de mamilos no DPN14. Os pesos relativos dos testículos expostos ao DBP estavam reduzidos apenas no DPN24. Animais expostos ao DBP mostraram níveis reduzidos de expressão de Pou5f1 e Mki67 no DPN24, e de Pou5f1 e Spry4 no DPN45. A exposição a AA reduziu a expressão de Pou5f1, Mki67 e Spry4, embora não significativamente. Nossos resultados sugerem que DBP atue reduzindo a proliferação celular e prejudi... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The increased incidence of testicular disorders in young men and the possible influence of environmental chemicals, such as dibutyl phthalate (DBP) and acrylamide (AA), requires experimental models for identifying modes of action. Most published reproductive toxicologic studies use RNA samples from the total testis to evaluate testicular gene expression; however, analyses of isolated cell types could provide a more specific tool. Among testicular germ cells, spermatogonia are critical since they represent the onset of spermatogenesis. This study aimed, 1) to establish a technique for spermatogonia isolation; 2) to apply this isolation technique to verify possible gene expression alterations (Pou5f1, Kitlg, Mki-67, Bak1 and Spry4) in prepubertal post-natal day, (PND24) and pubertal (PND45) testes after in utero and postnatal exposure to DBP or AA. The technique was efficient for isolation of a majority of spermatogonia. In utero DBP exposure led to reduced litter body weight at birth, reduced anogenital distance of male pups on PND4, and increased frequency of male nipple retention on PND14 compared to controls. DBP-exposed relative testes weights were reduced only at PND24 compared to control but they did not differ at PND45. DBP-exposed animals showed reduced expression levels of Pou5f1 and Mki67 on PND24, and reduced expression of Pou5f1 and Spry4 on PND45. AA exposure reduced expression of Pou5f1, Mki67 and Spry4 at PND45 although not significantly. Our results suggest tha... (Complete abstract click electronic access below) / Doutor

<p>The aim of this thesis was to verify the indicated occurrence of acrylamide formation in heating of food, to identify factors affecting the formation, and to identify important sources of acrylamide exposure from food. As a prerequisite for the studies, gas- and liquid-chromatographic methods with mass spectrometric detection were developed for the analysis of acrylamide in food. The developed methods showed a high correlation coefficient (0.99), high sensitivity and reproducibility. Acrylamide was demonstrated to occur in heated food products, with unexpectedly high levels in potato products (up to mg/kg level in potato crisps) and in beetroot. The identity of acrylamide was confirmed by the developed methods. </p><p>With potato as a food model, different factors affecting the acrylamide formation were tested. It was shown that the addition of asparagine and fructose, as well as heating temperature and time had a large impact on the formation. Other factors affecting the acrylamide content were pH, addition of other amino acids apart from asparagine, protein and other reducing sugars. No significant effects were observed from addition of neither antioxidant nor radical initiators. It was discovered that acrylamide could be formed during heating of biological materials similar to food, also at temperatures below 100 ˚C. Furthermore, it was demonstrated that a fraction of acrylamide evaporates during heating, similar to conditions for cooking in household kitchens, and during dry matter determinations in laboratories (65-130 ˚C). This constitutes an earlier unobserved source of exposure to acrylamide.</p><p>The method for extraction of food was studied with regard to yield of acrylamide. It was shown that the yield at pH ≥12 increases 3 - 4 times compared to normal water extraction for some foods products. Extraction at acidic pH or with enzymatic treatment was also tested, showing no effect on yield.</p><p>In a study with mice the bioaviability of acrylamide extracted with the normal water extration and at alkaline pH was compared. It was shown that the extra acrylamide released at alkaline pH gave insignificant contributions to the in vivo dose, measured by hemoglobin adducts.</p>

The aim of this thesis was to verify the indicated occurrence of acrylamide formation in heating of food, to identify factors affecting the formation, and to identify important sources of acrylamide exposure from food. As a prerequisite for the studies, gas- and liquid-chromatographic methods with mass spectrometric detection were developed for the analysis of acrylamide in food. The developed methods showed a high correlation coefficient (0.99), high sensitivity and reproducibility. Acrylamide was demonstrated to occur in heated food products, with unexpectedly high levels in potato products (up to mg/kg level in potato crisps) and in beetroot. The identity of acrylamide was confirmed by the developed methods. With potato as a food model, different factors affecting the acrylamide formation were tested. It was shown that the addition of asparagine and fructose, as well as heating temperature and time had a large impact on the formation. Other factors affecting the acrylamide content were pH, addition of other amino acids apart from asparagine, protein and other reducing sugars. No significant effects were observed from addition of neither antioxidant nor radical initiators. It was discovered that acrylamide could be formed during heating of biological materials similar to food, also at temperatures below 100 ˚C. Furthermore, it was demonstrated that a fraction of acrylamide evaporates during heating, similar to conditions for cooking in household kitchens, and during dry matter determinations in laboratories (65-130 ˚C). This constitutes an earlier unobserved source of exposure to acrylamide. The method for extraction of food was studied with regard to yield of acrylamide. It was shown that the yield at pH ≥12 increases 3 - 4 times compared to normal water extraction for some foods products. Extraction at acidic pH or with enzymatic treatment was also tested, showing no effect on yield. In a study with mice the bioaviability of acrylamide extracted with the normal water extration and at alkaline pH was compared. It was shown that the extra acrylamide released at alkaline pH gave insignificant contributions to the in vivo dose, measured by hemoglobin adducts.

published_or_final_version / Chemistry / Doctoral / Doctor of Philosophy

Thesis (Ph.D.) -- McMaster University, 1999. / Includes bibliographical references. Also available via World Wide Web.

Thesis (M.S.)--University of Michigan, 1970.

Thesis (M.S.)--University of Michigan, 1970.

Novel self-assembling copolymers of 4-Methacryloyloxyazobenzene and NN-dimethylacrylamide (MOAB-DMA) exhibiting a pronounced photoviscosity effects in water are described. An optimum polymer architecture corresponding to maximum contents of the azobenzene moieties that allowed for the aqueous solubility of the MOAB-DMA copolymer at ambient temperature was observed at a molar fraction of the MOAB moieties in the copolymer of 0.2. When blended with Pluronic F127 copolymers, the MOAB-DMA solutions become both irradiation- and temperature-sensitive and capable of sol-gel transitions depending on the MOAB-DMA content. Effect of the MOAB-DMA copolymer addition to concentrated Pluronic F127 solutions was studied by controlled stress rheology. The presence of the amphiphilic MOAB-DMA copolymers in the micellar Pluronic solutions enabled irradiation-dependent shifts in gelation temperature, while the viscoelastic gel modulus was not significantly affected by the MOAB-DMA copolymers, indicating that cubic lattice of the Pluronic responsible for the gel formation was not compromised. The UV- and T-dependent gelling media might prove to become viable electrophoretic separation matrices. / Singapore-MIT Alliance (SMA)