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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Re-engineering of the Duocarmycin Structural Architecture Enables Bioprecursor Development Targeting CYP1A1 and CYP2W1 for Biological Activity

Sheldrake, Helen M., Travica, S., Johansson, I., Loadman, Paul, Sutherland, Mark, Elsalem, Lina M.I., Illingworth, Nicola A., Cresswell, Alexander J., Reuillon, Tristan, Shnyder, Steven, Mkrtchian, S., Searcey, M., Ingelman-Sundberg, M., Patterson, Laurence H., Pors, Klaus 11 July 2013 (has links)
Yes / A library of duocarmycin bioprecursors based on the CPI and CBI scaffolds was synthesized and used to probe selective activation by cells expressing CYP1A1 and 2W1, CYPs known to be expressed in high frequency in some tumors. Several CPI-based compounds were pM–nM potent in CYP1A1 expressing cells. CYP2W1 was also shown to sensitize proliferating cells to several compounds, demonstrating its potential as a target for tumor selective activation of duocarmycin bioprecursors.
2

Smoking Cessation After Genotype Notification: Pilot Studies of Smokers Employed by a Municipal Government and Those on Nagoya University Medical Campus

Kano, Mayuko, Goto, Yasuyuki, Atsuta, Yoshiko, Naito, Mariko, Hamajima, Nobuyuki 10 1900 (has links)
No description available.
3

Análise do polimorfismo Mspl do gene CYP1A1m1 (Citocromo P450) em mulheres com endometriose

Souza, Suelene Ribeiro de 27 March 2012 (has links)
Made available in DSpace on 2016-08-10T10:38:35Z (GMT). No. of bitstreams: 1 Suelene Ribeiro de Souza.pdf: 13184565 bytes, checksum: 005b5afc9db66a26778e81d9edec24fa (MD5) Previous issue date: 2012-03-27 / Endometriosis is a disease that affects between 10 and 15% of women of reproductive age. Characterized by the presence of tissue resembling the endometrium similar to the uterine cavity outside the uterus, although some definitions specify that the ectopic tissue functional and sensitive to the action of hormones and their etiopathogenic mechanisms probably involve immunological abnormalities. The degree of involvement of endometriosis is based on the system proposed by the American Society for Reproductive Medicine (1985), based on the findings of laparoscopy. It is observed in the last 10 years, an increasing tendency to use a set of markers to detect changes induced by xenobiotics. Several genetic polymorphisms have been cited for the CYP1A1 gene, indicating a lack of functional protein that can cause an increase or a decrease in metabolic activity. The CYP1A1 gene encode the phase I enzymes involved in detoxification of estrogen metabolism, encodes an isoenzyme that catalyzes the oxidation of polycyclic aromatic hydrocarbons (PAHs) in phenolic compounds and epoxides. Mapped and located on the long arm of chromosome 15 (15q22-24). The aim of this study was to analyze the gene polymorphism frequency Mspl CYP1A1m1 with endometriosis. We analyzed 52 samples of peripheral blood of with endometriosis documented by laparoscopy (FÉRTILE) aged 25 to 35 women and 42 samples from women without endometriosis aged 25 to 57 years (control group). Molecular analysis by PCR (polymerase chain reaction). There was a statistically significant association (P = 0.039) between endometriosis and CYP1A1m1 polymorphic allele in women with endometriosis (32.70%) and compared with the control group 14,29%. We conclude that the gene polymorphism CYP1A1m1 correlates with endometriosis and polymorphisms are W1/m1candcm1/m1 morecfrequentcincpatientscwithcinfertilityandcwithvmorebseverebclinical bpicturebofbendometriosis. / Endometriose é uma enfermidade que afeta entre 10 e 15% das mulheres em idade reprodutiva. Caracterizada pela presença de tecidos semelhante a do endométrio, idênticos aos da cavidade uterina fora do útero. Apesar de algumas definições especificarem que o tecido ectópico funcional é sensível à ação de hormônios e seus mecanismos etiopatogênicos provavelmente envolvem anomalias imunológicas. O grau do comprometimento da endometriose é baseado no sistema proposto pela American Society for Reproductive Medicine (1985), com base nos achados de laparoscopia. Observa-se nos últimos 10 anos a crescente tendência em se utilizar um conjunto de marcadores para detectar alterações induzidas por xenobióticos. Inúmeros polimorfismos genéticos têm sido citados para o gene CYP1A1. O gene codifica enzimas da fase I envolvidas na desintoxicação no metabolismo de xenobióticos, codifica uma isoenzima que catalisa a oxidação de hidrocarbonetos aromáticos policíclicos (PAHs) em produtos fenólicos e epóxidos. Mapeado no e localizado no braço longo do cromossomo 15 (15q22-24). O objetivo do estudo foi analisar freqüência o polimorfismo do gene Mspl CYP1A1m1 com a endometriose. Foram analisadas 52 amostras de sangue periférico de mulheres com endometriose comprovadas por laparoscopia (FÉRTILE) com idades 25 a 35 anos mulheres e 42 amostras de mulheres sem endometriose com idades 25 a 57 anos (grupo controle). A análise molecular por meio da técnica da PCR (polymerase chain reaction). Constatou-se uma associação estatisticamente significativa (P= 0, 039) entre a endometriose e o alelo polimórfico m1 nas mulheres com endometriose (32,70%) quando comparadas ao grupo controle 14,29%. Concluiu-se que o polimorfismo m1 correlaciona-se com a endometriose e que os polimorfismos W1/m1 e m1/m1 estão mais freqüentes nas pacientes com infertilidade e com quadro clínico mais severo da endometriose.
4

Avaliação da presença do polimorfismo Msp1 da CYP1A1 do citocromo P-450 em mulheres assintomáticas portadoras de cisto mamário / Evaluation of the presence of Msp1 polymorphism of cytochrome P-450 CYP1A1 in asymptomatic women suffering from breast cyst

Fenile, Rogério [UNIFESP] 02 December 2010 (has links) (PDF)
Made available in DSpace on 2015-07-22T20:49:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-12-02. Added 1 bitstream(s) on 2015-08-11T03:26:14Z : No. of bitstreams: 1 Publico-12911.pdf: 1451194 bytes, checksum: 30932a96c61700c65652119d47fad0a7 (MD5) / Objetivo: o objetivo do estudo foi verificar a prevalência da alteração fibrocística da mama, na sua forma cística, dentre as diversas faixas etárias da população feminina e correlacionar a doença cística com a presença do polimorfismo Msp1 da CYP1A1 do citocromo P450 . Casuística e métodos: Estudo retrospectivo, caso-controle, desenvolvido entre março de 2005 a março de 2007. Submeteram-se a exame ultra-sonográfico, 204 mulheres, sendo divididas em dois grupos: 44 com doença cística mamária (casos) e 149 sem doenças mamárias (controles); 11 mulheres foram excluídas. Foi realizado estudo genético para a detecção do CYP1A1 através de reação em cadeia da polimerase e utilizado o teste de Fisher e c2 para a análise estatística. Resultados: A doença cística mamária diagnosticada pelo ultra-som esteve presente em 22% da população estudada. Os cistos eram do tipo simples em 93%, múltiplos em 75% dos casos, e mediam 4-10 mm em 46%. A mama esquerda no quadrante súpero-lateral foi a localização mais freqüente. O perfil epidemiológico-clínico dessas mulheres foi: branca, faixa etária de 41-50 anos, ciclos menstruais regulares, multípara e com queixa de mastalgia. Na análise genética do CYP1A1, observamos homozigoto selvagem numa freqüência de 68,2% no grupo casos e 66% no controle; heterozigotos 31,28% no grupo estudo e 26,8% no controle.Não houve aparecimento do homozigoto mutado no grupo casos surgindo como 7,2% no controle. Não houve diferença estatisticamente significante entre os grupos (p = 0,42). Conclusão: A prevalência e quase todo o perfil epidemiológico da doença cística mamária foram compatíveis com a literatura. Houve maior freqüência de heterozigotos mutados no grupo de casos, porém não de homozigotos.Não houve associação estatisticamente significante entre o polimorfismo da CYP1A1 e a doença fibrocística mamária na sua forma cística. / TEDE
5

Clonage de gènes de petits vertébrés susceptibles de voir leur expression induite par des pesticides environnementaux et séquençage et assemblage du génome de l'hirondelle bicolore (Tachycineta bicolor).

Doyon, Kathy January 2015 (has links)
Environ 3500 tonnes de pesticides sont étendues chaque année sur les terres agricoles du Québec. L’utilisation de plusieurs de ces substances a été interdite, car les ingrédients actifs qui les composaient avaient des effets toxiques sur les humains et/ou l’environnement. Malheureusement, certains qui sont toujours en vente ont aussi des effets secondaires non désirables. En effet, ces molécules exogènes ont le potentiel de moduler l’activation de protéines régulatrices comme le récepteur aux dioxines (AhR). AhR active, entre autres, l’expression de gènes faisant partie de la famille du cytochrome P450 (CYP1A1 et CYP1B1) qui sont impliqués dans la détoxification. Or, dans certains cas, ces enzymes mènent à la production de molécules mutagènes en augmentant la toxicité des ingrédients actifs en les métabolisant. Le projet global dans lequel s’insère ce projet de maîtrise vise à déterminer les effets génomiques des pesticides environnementaux sur des organismes vivants en milieu naturel dans les environs de la région de l’Estrie. Les espèces choisies comme modèles d’étude sont des insectivores, car les pesticides s’accumulent dans les lipides et que les insectes en sont une excellente source. Les consommateurs d’insectes sont donc d’excellents marqueurs du niveau de contamination de leur environnement par les pesticides. Les deux espèces sélectionnées sont l’hirondelle bicolore (Tachycineta bicolor) et la grande musaraigne (Blarina brevicauda). De façon plus spécifique, le projet de maîtrise se divise en deux principaux objectifs. Le premier objectif est de valider que les pesticides présents dans l’environnement sont en concentration suffisante pour modifier la régulation génétique d’animaux en milieu naturel. Cette validation se fera en comparant le taux d’expression de CYP1A1 et CYP1B1 (suite de leur activation par AhR) chez des bêtes ayant été exposées (ou non) à des pesticides. Comme le génome des deux modèles d’étude n’est pas encore séquencé, le clonage partiel des gènes à étudier (AhR et CYP1) a été entamé de même que la conception d’amorces qui permettra de quantifier le niveau d’expression de ces gènes par des réactions en chaîne par polymérase en temps réel (qPCR). À ce jour, une partie de la séquence d’AhR, de CYP1B1 et de trois gènes contrôles ont été séquencés pour l’hirondelle, ce qui a permis de concevoir des amorces pour la quantification de l’expression d’AhR et de deux contrôles. Pour la musaraigne, ce sont les gènes AhR, potentiellement CYP1A1 et trois contrôles qui ont été séquencés partiellement et ce sont les gènes AhR et les contrôles pour lesquels des amorces sont prêtes à être utilisées pour la quantification par qPCR. Le deuxième objectif est d’observer les effets génétiques des pesticides de manière globale sur des organismes vivants. Un génome de référence est donc nécessaire pour identifier les régions qui vont être régulées (directement ou indirectement) par les polluants d’origine agricole. Pour la grande musaraigne, c’est le génome de la musaraigne commune (Sorex araneus) qui sera utilisé, car ces deux espèces sont très proches phylogénétiquement. Par contre, pour l’hirondelle bicolore, le séquençage, l’assemblage et l’annotation de son génome seront essentiels parce que les espèces d’oiseaux actuellement séquencées sont trop éloignées pour permettre l’identification des régions qui seront régulées différemment en présence de pesticides. Le séquençage a été fait et l’assemblage a permis de couvrir 55% du génome de l’hirondelle bicolore. Cependant, il est très fractionné avec un N50 de 1339 et un peu plus de 600 000 contigs. Quelques étapes restent à accomplir pour optimiser l’assemblage tel que l’élimination de la contamination (estimée à 5%). L’annotation pourra être faite lorsque l’étape précédente sera finie. Compte tenu de l’ampleur du projet, ce qui a été effectué dans le cadre de la maîtrise contribuera grandement à la bonne continuation de celui-ci. Le projet global permettra de mieux comprendre l’impact des pesticides sur des organismes sauvages.
6

Mechanism Of Inhibition Of Cytochrome P4501a1 Associated 7-ethoxyresorufin O-deethylase (erod) Activity And Glutathione S-transferase (gst) Activities In Fish Liver By Phenolic Compounds/flavonoids

Yilmaz, Duygu 01 January 2010 (has links) (PDF)
Flavonoids, present in fruits, vegetables and beverages derived from plants, have been described as health-promoting, disease-preventing dietary supplements, and have activity as cancer preventive agents. The cancer protective effects of flavonoids have been attributed to a wide variety of mechanisms, including modulating enzyme activities resulting in the decreased carcinogenicity of xenobiotics. Cytochrome P4501A1 (CYP1A1) is a Phase I enzyme which is known to be involved in the activation of procarcinogens and Glutathione S-Transferase (GST) is a Phase II enzyme which is largely responsible for the detoxification of carcinogens. In this study, it was aimed to investigate the mechanisms of inhibition of CYP1A1 and GST activities of fish by phenolic compounds/flavonoids. Leaping mullet (Liza saliens), captured from highly polluted sites of izmir Bay, expressing high levels of CYP1A, were used in order to investigate these effects. It was demonstrated that all of the phenolic compounds/flavonoids used, exert an inhibitory effect on both CYP1A1 associated 7-Ethoxyresorufin-O-deethylase (EROD) activity and GST activities of fish, although the degree of inhibition was varied with the flavonoid used. Of the flavonoids tested, the most potent inhibitor of CYP1A1 associated EROD activity was found to be quercetin. The potency of the phenolic compounds/flavonoids to inhibit CYP1A1 associated EROD activity follow the sequence of quercetin &gt / resveratrol &gt / naringenin &gt / hesperidin &gt / rutin with IC50 values of 1.32 &micro / M, 3.59 &micro / M, 9.78 &micro / M, 98.5 &micro / M and 0.64 mM respectively. Quercetin, resveratrol, hesperidin and rutin were found to inhibit EROD activity in a competitive manner, on the other hand, naringenin was found to inhibit EROD activity in a non-competitive manner. Inhibition constant (Ki) values of quercetin, resveratrol, naringenin, hesperidin and rutin were calculated from Dixon plots as 0.12 &micro / M, 0.67 &micro / M, 2.63 &micro / M, 18 &micro / M and 0.1 mM, respectively. In the case of GST enzyme, it was demonstrated that all of the phenolic compounds/flavonoids used, exert an inhibitory effect on both total GST and GST-Mu activities of fish. Of the flavonoids tested, the most effective inhibitor of total GST activity was found to be resveratrol. The potency of the phenolic compounds/flavonoids to inhibit total GST activity follow the sequence of resveratrol &gt / quercetin &gt / rutin &gt / naringenin &gt / hesperidin with IC50 values of 7.1 &micro / M, 24.5 &micro / M, 89 &micro / M, 116 &micro / M and 118 &micro / M respectively. Resveratrol, quercetin and hesperidin were found to inhibit total GST activity in a competitive manner, on the other hand, rutin and naringenin were found to inhibit GST activity in a mixed type manner. Ki values of resveratrol, quercetin, hesperidin, naringenin and rutin were calculated from Dixon plots as 3.2 &micro / M, 12.5 &micro / M, 45 &micro / M, 128 &micro / M and 150 &micro / M respectively. In the case of GST-Mu activity, the most potent inhibitor was found to be rutin. The potency of the phenolic compounds/flavonoids to inhibit GST-Mu activity follow the sequence of rutin &gt / resveratrol &gt / quercetin &gt / naringenin &gt / hesperidin with IC50 values of 66.5 &micro / M, 72.3 &micro / M, 113.5 &micro / M, 135.5 &micro / M and 196 &micro / M, respectively. In conclusion, this study indicated that flavonoids were the strong inhibitors of CYP1A1 associated EROD activity and GST activities of mullet liver. The modulation of drug-metabolizing enzymes by flavonoids is important in terms of human health, since these enzymes can activate or inactivate carcinogens. The potential role of xenobiotic metabolizers CYP1 family in the activation of carcinogens and inactivation of chemotherapeutics suggests a potential therapeutic benefits in inhibiting these enzymes. The results of the present study support the hypothesis that flavonoids may be involved in the prevention of malignant transformation, by reducing the formation of carcinogens through inhibition of enzymes such as CYP1A1 which is known to be involved in carcinogen activation.
7

Atividade de etoxiresorufina-O-desetilase, freqüência de micronúcleos e níveis de metais em peixes da bacia do Rio Paraíba do Sul / Activity of etoxiresorufina-O-desetilase, frequency of micronucleated and levels of metals in fish of the Paraíba do Sul river

Santos, Laísa Maria Freire dos January 2004 (has links)
Made available in DSpace on 2012-09-06T01:11:42Z (GMT). No. of bitstreams: 2 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) 759.pdf: 1770109 bytes, checksum: 7d8d95d8da9a2b5c5db2f639195ddba2 (MD5) Previous issue date: 2004 / Neste estudo avaliamos a freqüência de micronúcleos no sangue periférico, a atividade da etoxiresorufina-O-desetilase - EROD (marcador da atividade enzimática da subfamília CYP1A) no fígado, e os níveis de metais no músculo de peixes acarás (Geophagus brasiliensis) e cascudos (Hypostomus affinis e Hypostomus luetkeni) coletados na bacia do rio Paraíba do Sul. Biomarcadores inespecíficos como Fator de Condição e Índice Hepático também foram utilizados. Os peixes foram capturados nos trechos superior, médio-superior, médio-inferior e inferior da bacia do rio Paraíba do Sul em dois períodos distintos (chuva e estiagem). Dentre as espécies estudadas, G. brasiliensis foi a que coletamos com maior freqüência ao longo de toda a bacia. G. brasiliensis, H. affinis e H. luetkeni apresentaram uma tendência de aumento dos fatores de condição no período de chuvas quando comparamos aos valores obtidos no período de estiagem. Volta Redonda foi a região em que se coletou G. brasiliensis com os maiores valores de índice hepático. A freqüência de micronúcleos nos eritrócitos do sangue periférico das três espécies foi muito baixa (0,00-2,00 ) evidenciando ausência de dano genotóxico. A atividade de EROD em G. brasiliensis foi maior do que a das outras duas espécies em todos os locais de coleta. Na região de Levy Gasparian, onde foi possível capturar um grande número de peixes, a atividade de EROD em G. brasiliensis machos foi maior do que em fêmeas. A atividade de EROD de G. brasiliensis variou entre os locais de coleta, sendo os maiores valores registrados nos peixes de São Luiz do Paraitinga, Barra Mansa, Levy Gasparian, Três Rios e Sapucaia e os menores nos peixes de Paraibuna, Italva, Rio das Flores, Santo Antônio de Pádua e Campos. Os níveis de metais foram baixos nas três espécies em todas as regiões e no caso de chumbo e mercúrio, inferiores aos valores máximos permitidos pela regulamentação brasileira de alimentos (ANVISA). Estudos devem ser realizados para investigar as diferenças de EROD entre G. brasiliensis, H. affinis e H. luetkeni. As regiões de São Luiz do Paraitinga, Barra Mansa, Três Rios, Levy Gasparian e Sapucaia devem ser melhor investigadas quanto a poluentes indutores de CYP1A, uma vez que os peixes coletados nestes locais apresentaram as mais elevadas atividades de EROD.
8

Avaliação da expressão dos genes cFOS, IL-1b, CYP1a1 e CYP1b1 em Danio rerio expostos a Benzo[a]pireno e tratados com ligantes do receptor P2X7 / Gene expression evaluation of cFOS, IL-1, CYP1a1 and CYP1b1 in Danio rerio exposed to Benzo[a]pyrene and treated with P2X7 receptor ligands

Chamelete, André [UNESP] 25 January 2016 (has links)
Submitted by André Chamelete null (andre_ecco@hotmail.com) on 2016-02-12T14:02:00Z No. of bitstreams: 1 Dissertação de Mestrado - FINAL.pdf: 663322 bytes, checksum: 5ca648d67a3a798d08f9c68653ac3ca2 (MD5) / Approved for entry into archive by Sandra Manzano de Almeida (smanzano@marilia.unesp.br) on 2016-02-12T17:20:57Z (GMT) No. of bitstreams: 1 chamelete_a_me_sjrp.pdf: 663322 bytes, checksum: 5ca648d67a3a798d08f9c68653ac3ca2 (MD5) / Made available in DSpace on 2016-02-12T17:20:57Z (GMT). No. of bitstreams: 1 chamelete_a_me_sjrp.pdf: 663322 bytes, checksum: 5ca648d67a3a798d08f9c68653ac3ca2 (MD5) Previous issue date: 2016-01-25 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / O BaP é um contaminante ambiental capaz de causar inflamação e desregulação de vias celulares. Pela ação da CYP1a1 e CYP1b1, é convertido a metabólitos mais reativos. A literatura mostra que o BaP aumenta a expressão de algumas citocinas próinflamatórias, como a IL-1, porém, são bem contraditórios os relatos sobre o efeito do BaP no cFOS, o qual apresenta papel importante na proliferação, na formação de tumores e, possivelmente, na inflamação. O objetivo deste estudo foi de elucidar a participação do receptor purinérgico P2X7 sobre a expressão dos genes IL-1 e cFOS, durante exposição ao BaP. Foi empregado as técnicas de qPCR para quantificação de expressão gênica, e testes de correlação e regressão entre IL-1 e cFOS. A exposição ao BaP induziu a expressão dos dois genes, além das enzimas do seu metabolismo. Quando bloqueado o receptor P2X7, além de uma menor indução das CYPs, os níveis de IL-1 e cFOS caíram abaixo dos níveis controle, sugerindo a participação do P2X7. Os testes de correlação e regressão mostraram uma relação forte direta entre IL-1 e cFOS, reforçando o papel do cFOS na inflamação. / BaP is an environmental contaminant capable to cause inflammation and impair cellular pathways. CYP1a1 and CYP1b1 convert it to more reactive metabolites. Studies show that BaP enhances some proinflammatory citokines expression, like IL-1, yet reports about BaP affecting cFOS, which plays important role in proliferation, tumor formation and inflammation, are controversial. This work aimed to elucidate whether P2X7 purinergic receptor plays a role in IL-1 and cFOS expression during BaP exposure. We applied qPCR techniques to quantify gene expression, correlation and regression assays. Our results showed that BaP raised both IL-1 and cFOS genes expression, besides CYPs ones. Morevoer, when blocking P2X7 receptor, IL-1 and cFOS expression dropped under normal levels, which suggest P2X7 participation, in addition to a smaller enzymes induction. Correlation and regression assays exhibited a strong straight relationship between IL-1 and cFOS expression, reinforcing the role of cFOS in inflammation.
9

Development of an in vitro mechanistic toxicity screening model using cultured hepatocytes

Van Tonder, Jacob John 26 April 2012 (has links)
In vitro testing includes both cell-based and cell-free systems that can be used to detect toxicity induced by xenobiotics. In vitro methods are especially useful in rapidly gathering intelligence regarding the toxicity of compounds for which none is available such as new chemical entities developed in the pharmaceutical industry. In addition to this, in vitro investigations are invaluable in providing information concerning mechanisms of toxicity of xenobiotics. This type of toxicity testing has gained popularity among the research and development community because of a number of advantages such as scalability to high throughput screening, cost-effectiveness and predictive power. Hepatotoxicity is one of the major causes of drug attrition and the high cost associated with drug development poses a heavy burden on the development of new chemical entities. Early detection of hepatotoxic agents by in vitro methods will improve lead optimisation and decrease the cost of drug development and reduce drug-induced liver injury. Literature highlights the need for a cellbased in vitro model that is capable of assessing multiple toxicity parameters, which assesses a wider scope of toxicity and would be able to detect subtle types of hepatotoxicity. The present study was aimed at developing an in vitro procedure capable of mechanistically profiling the effects of known hepatotoxin dichlorodiphenyl trichloroethane (DDT) and its metabolites, dichlorodiphenyl dichloroethylene (DDE) and dichlorodiphenyl dichloroethane (DDD) on an established liver-derived cell line, HepG2, by evaluating several different aspects of cellular function using a number of simultaneous in vitro assays on a single 96 well microplate. Examined parameters have been suggested by the European Medicines Agency and include: cell viability, phase I metabolism, oxidative stress, mitochondrial toxicity and mode of cell death (apoptosis vs. necrosis). To further assess whether the developed method was capable of detecting hepatoprotection, the effect of the known hepatoprotectant, N-acetylcysteine, was determined. Viability decreased in a dose-dependent manner yielding IC50 values of 54 μM, 64 μM and 44 μM for DDT, DDE and DDD, respectively. Evaluation of phase I metabolism showed that cytochrome P4501A1 activity was dose-dependently induced. Test compounds decreasedlevels of reactive oxygen species, and significantly hyperpolarised the mitochondrialmembrane potential. Assessment of the mode of cell death revealed a significant elevation of caspase-3 activity, with DDD proving to be most potent. DDT alone induced dosedependent loss of membrane integrity. These results suggest that the tested compounds produce apoptotic death likely due to mitochondrial toxicity with subsequent caspase-3 activation and apoptotic cell death. The developed in vitro assay method reduces the time it would take to assess the tested parameters separately, produces results from multiple endpoints that broadens the scope of toxicity compared to single-endpoint methods. In addition to this the method provides results that are truly comparable as all of the assays utilise the same batch of cells and are conducted on the same plate under the exact same conditions, which eliminates a considerable amount of variability that would be unavoidable otherwise. The present study laid a solid foundation for further development of this method by highlighting the unforeseen shortcomings that can be adjusted to improve scalability and predictive power. / Thesis (PhD)--University of Pretoria, 2011. / Pharmacology / unrestricted
10

A Test of the Hypothesis That Environmental Chemicals Interfere With Thyroid Hormone Action in Human Placenta

Geromini, Katherine 01 January 2012 (has links) (PDF)
Thyroid hormone is essential for normal brain development and recognition of this has led to universal screening of newborns for thyroid function to ensure that circulating levels of thyroid hormone are within a range known to be supportive of normal growth and mental development. Environmental chemicals that interfere with thyroid function are known to inhibit normal growth and mental development. Work from our lab and from labs internationally demonstrates in animal systems that some industrial chemicals such as PCBs, PBDEs, and others may interact with the thyroid hormone receptor(s) in ways that are not predicted by changes in serum thyroid hormone levels. Our work demonstrates that the enzyme CYP1A1 must metabolize some individual PCB congeners before they can interact with the thyroid receptor. In animals, this requirement appears to be manifested in part by a strong correlation between CYP1A1 and TH target gene expression. Here we present that this pattern extends to humans by demonstrating a correlation between increased CYP1A1 mRNA and an abundance of thyroid hormone responsive gene mRNA.

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