A alteração do clima da Terra induzida pelas atividades humanas e a poluição têm sido apontados como potenciais estressores sinergísticos que ameaçam a persistência de populações de anuros em ambientes naturais. Entretanto, mediante um cenário complexo de mudanças antropogênicas globais, o conhecimento acerca do efeito isolado de estressores, bem como de sua interação, é ainda incipiente. Além do aquecimento como tendência central, a mudança climática tem sido relacionada, de modo cada vez mais consistente, ao aumento de eventos extremos, que podem resultar no aumento da ocorrência e da magnitude de temperaturas diárias extremas quentes e frias, impondo desafios adicionais à biota. No presente trabalho, foi testada a hipótese de que o aquecimento e a flutuação termal diária interferem na toxicidade da atrazina sobre larvas de três espécies de anuros (Dendropsophus minutus, Leptodactylus podicipinus e Physalaemus nattereri) que habitam áreas de cultivo de cana-de-açúcar, onde a utilização desse herbicida é frequente. Para testar essa hipótese, foram estudados os efeitos do aquecimento (23-27 ºC para D. minutus e L. podicipinus e 28-32 ºC para P. nattereri), da flutuação termal diária (23±4 ºC e 27±4 ºC para D. minutus e L. podicipinus e 28±4 ºC para P. nattereri) e da exposição à atrazina (120 μg/L para D. minutus e L. podicipinus e 30, 120 e 180 μg/L para P. nattereri), de maneira isolada e combinada, no desenvolvimento larval dessas espécies. A fim de compreender tais efeitos, foram monitorados os seguintes parâmetros: tempo de desenvolvimento, massa e comprimento na metamorfose, condição corpórea, proteínas e lipídios hepáticos totais, atividade de enzimas antioxidantes (catalase, GST e SOD) e nível de peroxidação lipídica. As respostas encontradas foram diferentes entre as funções de desempenho fisiológico analisadas, com possíveis consequências positivas ou negativas para a aptidão, além de terem sido espécie-específicas. Além disso, no caso do efeito combinado dos estressores analisados, foram observadas interações sinergísticas, neutras ou mesmo antagônicas. De modo isolado, a atrazina e a flutuação termal promoveram o retardo ou a aceleração do desenvolvimento, enquanto o aquecimento acelerou ou não teve efeito sobre o mesmo. A atrazina, surpreendentemente, promoveu a melhora da condição corpórea, enquanto a flutuação termal diária promoveu a melhora, a piora ou não teve efeito sobre esse parâmetro, e, por fim, o aquecimento piorou ou não alterou a condição corpórea. Os três fatores testados, de modo isolado, promoveram a ativação da catalase e, de modo menos frequente, da GST, embora apenas o aquecimento, de modo isolado, tenha induzido o dano oxidativo, com aumento do nível de LPO tendo sido observado na espécie D. minutus. Os fatores analisados (aquecimento, flutuação termal diária e exposição a concentrações ecologicamente relevantes de atrazina) tiveram efeito sinergístico em D. minutus, em que houve piora da condição corpórea, e neutro em L. podicipinus, espécie na qual não foi verificada alteração nesse parâmetro. Houve ainda sinergismo na indução de dano oxidativo (aumento do nível de LPO), constatado nas três espécies analisadas nos casos mais severos de combinação de estressores. O presente trabalho evidencia a complexidade da interação de fatores ambientais e, portanto, reafirma a necessidade de estudar os efeitos das mudanças globais sobre a biota para a melhor definição dos limites de tolerância e da capacidade de resiliência das espécies / Global climate change, induced by human activities, and pollution has been considered as potential synergistic stressors that threaten the persistence of amphibian populations in natural environments. However, considering the complexity of anthropogenic global changes, the current state of knowledge on the isolated effects of these variables, and their interactions, is still incipient. Besides the tendency to average global warming, climate change has been consistently associated to enhanced extreme events, which may result in an increase of frequency and magnitude of extreme hot and cold daily temperatures, imposing additional challenges to the biota. In this study, we tested the hypothesis that the increase in temperature and daily thermal fluctuation intensify the toxicity of atrazine on larvae of three anuran species (Dendropsophus minutus, Leptodactylus podicipinus and Physalaemus nattereri), which inhabit industrial sugarcane cultures areas, where the use of this herbicide is frequent. To test this hypothesis, the effects of the increase in temperature (23-27 °C for D. minutus and L. podicipinus, and 28-32 °C for P. nattereri), the daily thermal fluctuation (23 ± 4 °C and 27 ± 4 °C for D. minutus and L. podicipinus, and 28 ± 4 °C for P. nattereri), and the exposition to atrazine (120 μg/L for D. minutus and L. podicipinus, and 30, 120 and 180 μg/L for P. nattereri) on the larval development of these species were studied in an isolated and in a combined way. To evaluate the effects of these stressors, time of metamorphosis, mass and length, body condition, total hepatic proteins and lipids, antioxidant enzymes (catalase, GST and SOD) activity, and lipid peroxidation (LPO) were monitored. The responses were variable, which possible positive or negative consequences to fitness, according to the endpoint, and were species-specific. Moreover, the interactive effects of these stressors were synergistic, antagonistic, and even neutral. Atrazine and thermal fluctuation led to a delay or acceleration of the metamorphosis, while the warming accelerated or did not affect development. Surprisingly, exposure to atrazine improved body condition, whereas the daily thermal fluctuation improved, worsened or did not affect this parameter, and, finally, the increase of temperature worsened or did not affect body condition. All stressors promoted the activation of catalase, and, less frequently, activated GST. However, the increase in temperature was the only isolated factor to induce oxidative damage as observed in D. minutus. The factors studied here (increasing in temperature, daily thermal fluctuation and exposure to environmentally relevant concentrations of atrazine) had synergistic effect on the induction of oxidative damage in the all species analyzed. This study highlights the complexity of interaction among stressors and, therefore, reaffirms the need to study the effects of environmental changes on the biota to better understand the limits of tolerance and resilience of the species

Made available in DSpace on 2016-08-10T10:39:25Z (GMT). No. of bitstreams: 1 Ana Carolina Franco Finotti.pdf: 1174608 bytes, checksum: 94099b94b18d65f4bd321975ff4f8b0e (MD5) Previous issue date: 2009-06-19 / The infertility is a significant problem in the world-wide population, more than affecting 15% of the couples in reproductive age. Separately the man contributes with 50% in the conjugal infertility. The genetic causes are responsible for 60% of the cases of idiopathic infertility. The polymorphisms in genes that codify enzymes of phase II detoxificant can modify its expression or function, modifying the biometabolism of toxic composites to the male reproductive system. Genes GSTM1 and GSTT1 they codify enzymes of same name, they are basic in the process of detoxification of the endogenous or exogenous xenobiotics, facilitating its excretion. The aim of the present study was to detect the polymorphism of genes GSTM1 and GSTT1 in men with idiopathic infertility for the Service of Reproduction Human being of the Hospital das Clínicas de Goiânia (HC), according to finding of the spermograms. To the total 304 samples of patients with age between 15 had been evaluated - 69 years, diagnosis with idiopathic infertility, in the period of 2004 - 2006. The results had observed in samples of DNA of semen gen frequencies of 28,6% (30/105) for genotype GSTM1/T1 (null) in normal individuals and 50,0% (6/128) for the GSTM1/T1 (null) in abnormal individuals, demonstrating to significant relation statistics enters the polymorphism of gene GSTM1 and GSTT1 with the idiopathic male infertility. The seminal alteration associate to the polymorphic genotype of higher frequency GSTM1/T1 (null) does the number alteration (oligozoospermia) possessing bigger frequency in such a way for GSTM1 (78.9%) as well as for GSTT1 (73.7%), although not to be significant. Analyzing the genotype frequencies of the samples of blood we do not find statistical associations with male infertility, since 46.6% (34/76) are GSTM1/T1 (null) in normal individuals and 58, 4% (73/125) are GSTM1/T1 (null) in abnormal individuals. Significant influence of the consumption of alcohol, cigarette, mumps and use of xenobiotics analyzed in the basic parameters of the analyzed spermogram was not found. Polymorphic individuals for genes GSTM1 and GSTT1 are susceptiveis the reduction in the seminal quality and infertility, possibly being that oligospermic individuals are affected by the genic polymorphism. / A infertilidade é um problema significante na população mundial, afetando mais de 15% dos casais em idade reprodutiva. Isoladamente o homem contribui com 50% na infertilidade conjugal. As causas genéticas são responsáveis por 60% dos casos de infertilidade idiopática. O polimorfismo em genes que codificam enzimas detoxificantes de fase II pode alterar sua expressão ou função, modificando o biometabolismo de compostos tóxicos ao sistema reprodutor masculino. Os genes GSTM1 e GSTT1 codificam enzimas de mesmo nome, são fundamentais no processo de detoxificação dos xenobióticos endógenos ou exógenos, facilitando sua excreção. O objetivo principal do presente estudo foi detectar o polimorfismo dos genes GSTM1 e GSTT1 em homens com infertilidade idiopática pelo Serviço de Reprodução Humana do Hospital das Clínicas de Goiânia (HC), segundo o laudo dos espermogramas. Ao total foram avaliadas 304 amostras de pacientes com idade entre 15 e 69 anos, diagnosticados com infertilidade idiopática, no período de 2004 a 2006. Os resultados observados em amostras de DNA de sêmen, freqüências genotípicas de 28,6% (30/105) para o genótipo GSTM1/T1(nulo) em indivíduos normais e 50% (6/128) para o GSTM1/T1(nulo) em indivíduos alterados, demonstrando significativa relação estatística entre o polimorfismo do gene GSTM1 e GSTT1 com a infertilidade masculina idiopática. A alteração seminal associada ao genótipo polimórfico de maior freqüência GSTM1/T1 (nulo) é a alteração de número (oligozoospermia) possuindo maior freqüência tanto para GSTM1 (nulo) (78,9%) quanto GSTT1 (nulo) (73,7%), apesar de não ser significante. Analisando as freqüências genotípicas das amostras de sangue não encontramos associações estatísticas com a infertilidade masculina, já que 46,6% (34/76) são GSTM1/T1 (nulo) em indivíduos normais e 58, 4% (73/125) são GSTM1/T1 (nulo) em indivíduos alterados. Não foi encontrada influência significativa do consumo de álcool, cigarro, caxumba e uso de xenobióticos analisados nos parâmetros básicos dos espermogramas analisados. Indivíduos polimórficos para os genes GSTM1 e GSTT1 estão mais susceptíveis a redução na qualidade seminal e possivelmente infertilidade, sendo que indivíduos oligospérmicos são os mais afetados pelo polimorfismo gênico.

The non-alcoholic fatty liver disease (NAFLD) is the most common liver pathology in developed countries with an estimated prevalence of 20 to 30% in the American population. A typically benign and asymptomatic pathology, NAFLD is characterized by hepatic steatosis and abnormal levels of hepatic enzymes stemming from an increase in circulating free fatty acids originating from white adipose tissue lipolysis, an increased de novo lipogenesis, reduced fatty acid oxidation and decreased hepatic triglycerides secretion, all within an insulin resistance context. NAFLD has the potential to progress to the non-alcoholic steatohepatitis (NASH), a condition marked by inflammation, advanced oxidative stress and fibrosis. NASH is expected to be the leading cause of liver transplant by 2020 due to its complications (i.e.: cirrhosis, hepatocellular carcinoma and liver failure). Various xenobiotics such as pesticides have been shown to promote the apparition and development of NAFLD. Of interest to this study is the neonicotinoid imidacloprid, more contemporarily known for its suspected role in the colony collapse disorder of various anthophilae species. Imidacloprid has been shown to induce hepatic oxidative stress in rats, a significant factor in the development of NAFLD and its progression to NASH. Lifestyle modifications, namely physical exercise, is a current treatment which has been proven beneficial to prevent and treat NAFLD by reducing hepatic steatosis, oxidative stress and improving insulin sensitivity. The role of any neonicotinoid on the development of NAFLD has yet to be examine and few have looked at the role of exercise in the treatment of NAFLD brought about by pesticide contamination.

<p>Prisustvo organskih zagađujućih supstanci (farmaceutski aktivnih komponenata i prirodnih toksina‐mikotoksina) u uzorcima životne sredine i namirnicama je u porastu kao posledica novih industrijskih procesa i ostalih antropogenih aktivnosti, kao i klimatskih promena. Takođe veliku pažnju javnosti privlače i neorganske zagađujuće supstance kao &scaron;to su te&scaron;ki elementi. S obzirom da zagađujuće supstance imaju negativan uticaj na životnu sredinu i zdravlje ljudi, u svetu se preduzimaju mere u cilju smanjenja stepena izloženosti toksičnim jedinjenjma i posledicama izlaganja. Trenutno, jedan od najvećih izazova, jeste procena rizika povezana sa velikim brojem zagađujućih supstanci u tragovima ili u tzv. ultratragovima, uključujući &ldquo;novo&rdquo; otkrivena zagađujuća jedinjenja, a jedan od osnovnih trendova je razvoj i primena brzih i efikasnih metoda za njihovu analizu u ispitivanim uzorcima na bazi naprednih hromatografskih i spektrometrijskih tehnika.<br />Tehnike bazirane na tečnoj hromatografiji sa različitim masenim analizatorima za<br />kvantifikaciju organskih zagađujućih supstanci kao i metode zasnovane na atomskoj<br />apsorpcijonoj spektrometriji za određivanje ultratragova neorganskih zagađujućih supstanci postale su referentne na međunarodnom nivou. Ovakve napredne spregnute tehnike postale su važne za identifikaciju, kvantifikaciju i praćenje različitih zagađujućih supstanci u uzorcima životne sredine i namirnicama i proceni njihovog &scaron;tetenog uticaja na zdravlje čoveka. S obzirom da su u literaturi retka istraživanja koja se bave razvojem i primenom metoda zasnovanih na naprednim hromatografskim i spektrometrijskim tehnikama i određivanju organskih i neorganskih zagađujućih supstanci u matriksima životne sredine i namirnicama sa prostora zapadnog Balkana, a uzimajući u obzir njihovu važnost, specifični ciljevi disertacije su:<br />&bull; unutra&scaron;nja (&bdquo;inhouse&ldquo;)<br />provera kvaliteta i pouzdanosti postojeće &bdquo;multi‐rezidualne&ldquo;<br />metode zasnovane na UHPLC‐QqLIT‐MS/MS za analizu 81‐e farmaceutski aktivne<br />komponente (PhAC) u otpadnoj, povr&scaron;inskoj, podzemnoj i pijaćoj vodi i po prvi put<br />dobijanje sveobuhvatnih rezultata njihovog prisustva u različitim tipovima vode sa<br />područja Srbije;<br />&bull; unutra&scaron;nja (&bdquo;inhouse&ldquo;) provera kvaliteta i pouzdanosti postojeće &bdquo;multi‐toksin&ldquo;<br />metode za analizu 8 Fusarium mikotoksina u uzorcima ozime p&scaron;enice različitih sorti<br />zasnovane na HPLC‐QqQ‐MS/MS radi određivanja regionalnih razlika između žitnih<br />regiona kao i otpornosti ispitivanih sorti p&scaron;enice na kontaminaciju Fusarium<br />toksinima;<br />&bull; modifikacija postojeće &bdquo;multi‐toksin&ldquo; metode zasnovane na UHPLC‐QqQ‐MS/MS za<br />analizu 11 osnovnih mikotoksina u uzorcima bra&scaron;na i njena unutra&scaron;nja (&bdquo;inhouse&ldquo;)<br />provera kvaliteta i pouzdanosti, kao i provera kroz interlaboratorijsko poređenje,<br />radi dobijanja podataka za procenu &scaron;tetnog uticaja ispitivanih mikotoksina na<br />zdravlje populacije;<br />&bull; razvoj &bdquo;multi‐toksin&ldquo; (vi&scaron;ekomponentne) i &bdquo;multi‐matriks&ldquo; (za vi&scaron;e matriksa) metode<br />bazirane na UHPLC‐QqQ‐MS/MS za analizu 10 mikotoksina u različitim vrstama<br />ko&scaron;tuničavog voća čija provera kvaliteta je zasnovana na intralaboratorijskoj proveri<br />tačnosti i preciznosti dobijenih rezultata;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; &bull; primena postojeće analitičke procedure zasnovane na naprednoj tehnici pripreme<br />(mikrotalasnoj digestiji) različitih uzoraka biljnog i životinjskog porekla i provera<br />kvaliteta metode identifikacije i kvantifikacije zasnovane na atomskom apsorpcionom spektrometru sa grafitnom kivetom (GFAAS) radi dobijanja sveobuhavatnih rezultata o prisustvu te&scaron;kih elemenata (arsena, olova i kadmijuma) radi procene izloženosti stanovni&scaron;tva Srbije toksičnim neorganskim elementima.<br />Postignuti rezultati predstavljaju jedinstvene rezultate za područje Srbije dobijene<br />primenom naprednih spregnutih tehnika koje imaju značajnu ulogu u praćenju prisustva većeg broja organskih i neorganskih zagađujućih supstanci u izabranim uzorcima životne sredine i namirnica, (regulisanih postojećim zakonodavstvom) radi procene stepena zagađenosti ili u slučaju jedinjenja koja nisu regulisana zakonodavstvom radi sticanja novih saznanja o njihovom prisustvu i proceni mogućeg negativnog uticaja na životnu sredinu i zdravlje populacije.</p> / <p>The presence of organic pollutants in environmental samples and food (pharmaceutically active components and natural toxins‐mycotoxins) is increased as a result of new industrial processes and other anthropogenic activities, as well as climate change. Similarly heavy elements as inorganic pollutants have attracted worldwide attention. Since, these pollutants have negative impact on environment and human health, extremely efforts are undertaken in the world to reduce the level of exposure to these pollutants and consequences of the exposure. Currently, one of the highest challenges is to assess the risk associated with a large number of pollutants in trace or ultra trace levels, including &quot;new&quot; (emerging) discovered pollutants, and one of the main trends is development and implementation of fast and efficient methods for their analysis on the basis of advanced chromatographic and spectrometric techniques. Therefore, coupled techniques have become important for the identification, quantification and monitoring of various pollutants in environmental samples and food and assessment of their hazard impact on human health. Since, there are scarce data about the development and application of advanced methods based on chromatographic and spectrometric techniques for determination of organic and inorganic pollutants in environmental matrices and food from the Western Balkan, and taking into account their importance, specific objectives of<br />the dissertation were: &bull; internal (&quot;in‐house&quot;) quality control of the existing &quot;multi‐residual&quot; method based on UHPLC‐QqLIT‐MS/MS for analysis of 81 pharmaceutically active components (PhAC) in wastewater, surface, underground and drinking water due to obtained for the first time comprehensive results of their presence in different types of water from Serbia; &bull; internal (&quot;in‐house&quot;) quality control of the existing &quot;multi‐toxin&quot; method for the analysis of 8 Fusarium mycotoxins in samples of different winter wheat cultivars based on HPLC‐QqQ‐MS/MS to determine the differences among wheat-growing regions as well as the resistance of the analysed wheat cultivars towards Fusarium toxins; &bull; modification of existing &quot;multi‐toxin&quot; method based on UHPLC‐QqQ‐MS/MS for analysis of 11 principal mycotoxins in samples of flour and its internal (&quot;in‐house&quot;) quality control as well as verification through the interlaboratory comparison, in order to obtain data for assessing the hazard effect of these mycotoxins on the health of the population; &bull; the development of &quot;multi‐toxin&quot; and &quot;multi‐matrix&quot; method based on UHPLC‐QqQMS/ MS for the analysis of 10 mycotoxins in various types of nuts based on intralaboratory verification of the accuracy and precision of the obtained results; &bull; application of analytical procedure based on advanced preparation technique (microwave digestion) and atomic absorption spectrometer with a graphite furnace (GFAAS) and its verification in order to obtain comprehensive results on the presence of heavy elements (arsenic, lead and cadmium) in different samples of plant and animal origin to assess the exposure of the Serbian population to toxic inorganic elements. The obtained results are unique for the Serbia. They are obtained by applying advanced coupled techniques that have a significant role in monitoring the presence of a numerous organic and inorganic pollutants in analyzed samples of the environment and food. The presented results contribute to the assessment of pollution degree and in the case of new (emerging) not regulated polutant they might give new information about the possible negative impact on the environment and health of the population.</p>

Submitted by Suzana Dias (suzana.dias@famerp.br) on 2018-10-19T18:22:34Z No. of bitstreams: 1 StephaniePiacentidosSantos_dissert.pdf: 499357 bytes, checksum: e7f6872448bfb8c9bb786b4bd30f9c94 (MD5) / Made available in DSpace on 2018-10-19T18:22:34Z (GMT). No. of bitstreams: 1 StephaniePiacentidosSantos_dissert.pdf: 499357 bytes, checksum: e7f6872448bfb8c9bb786b4bd30f9c94 (MD5) Previous issue date: 2016-11-08 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Breast cancer is a leading cause of death in women worldwide. The etiology of this disease is multifactorial, including factors such as habits and lifestyle, hormonal, environmental and genetic. The xenobiotic metabolism contribute to the development of breast carcinoma and polymorphisms in genes encoding enzymes in this pathway, such as GST (GSTM1 and GSTT1) and CYPs (CYP1A1*2A and CYP1A1*2C) have been associated to breast cancer. Objectives: To investigate the influence of the GSTM1, GSTT1, CYP1A1*2A and CYP1A1*2C polymorphisms at the risk for developing breast cancer; to evaluate the association of polymorphisms and risk factors (age, smoking, alcohol, clinical features), tumor clinical and histopathologic features in breast cancer. Methods: This case-control study included 752 women, 219 patients and 533 controls. Molecular analysis were performed by PCR-multiplex (GSTM1 null and GSTT1 null), PCR-RFLP (CYP1A1*2A) and real-time PCR (CYP1A1*2C). For the statistical analysis, the MINITAB 16.0 (Multiple Logistic Regression Test), SNPstats (Inheritance Model tests) and BioEstat 5.0 (Hardy-Weinberg test) tests were used. Results: Women with old age and the alcohol consumption had increased risk for developing breast cancer. Women with breast cancer had slightly higher frequency (51%) of the GSTM1 null genotype than women without cancer (49%), however this difference was not significant statistically. GSTT1 null genotype was also not associated with breast cancer. CYP1A1*2A polymorphism was associated with the risk for breast cancer and CYP1A1*2C polymorphism was more frequent in tumors with no distant metastases. Conclusions: Women with age advanced and who drink alcohol present increased risk for developing breast cancer. CYP1A1*2A polymorphism is associated with breast cancer and CYP1A1*2C polymorphism is related to women with no distant metastases. The polymorphisms of the GSTM1 and GSTT1 genes are not associated with the development of breast cancer. / O câncer de mama é uma das principais causas de morte em mulheres no mundo. A etiologia desta doença é multifatorial e inclui fatores como hábitos, estilo de vida, hormonais, ambientais e genéticos. O metabolismo de xenobióticos contribui para o desenvolvimento do carcinoma mamário, e polimorfismos nos genes que codificam enzimas desta via, tais como GSTs (GSTM1 e GSTT1) e CYPs (CYP1A1*2A e CYP1A1*2C) têm sido associados ao câncer de mama. Objetivos: Investigar a influência dos polimorfismos GSTM1, GSTT1, CYP1A1*2A e CYP1A1*2C no risco de desenvolvimento de câncer de mama; avaliar a associação dos polimorfismos e fatores de risco (idade, fumo, álcool, características clínicas) no câncer de mama, assim como características clínicas e histopatológicas do tumor. Casuística e Métodos: O presente estudo caso-controle incluiu 752 mulheres, 219 pacientes e 533 controles. Para análise molecular foram realizadas as técnicas de PCR-Multiplex (GSTM1 e GSTT1), PCR-RFLP (CYP1A1*2A) e PCR em tempo real (CYP1A1*2C). Para a análise estatística foram utilizados os programas MINITAB 16.0 (teste de Regressão Logística Múltipla), SNPstats (testes de Modelos de Herança) e BioEstat 5.0 (teste de Equílibrio de Hardy-Weinberg). Resultados: Mulheres com idade avançada e com o hábito etilista apresentaram risco aumentado para o desenvolvimento de câncer de mama. Mulheres com carcinoma mamário apresentaram frequência discretamente maior (51%) do genótipo nulo GSTM1 em relação a mulheres sem câncer (49%), no entanto essa diferença não foi estatisticamente significante. O genótipo nulo de GSTT1 também não foi associado ao câncer de mama. O polimorfismo CYP1A1*2A foi associado ao risco para câncer de mama e o polimorfismo CYP1A1*2C foi mais frequente em tumores com ausência de metástase à distância. Conclusões: Mulheres com idade avançada e que ingerem bebida alcoólica apresentam risco aumentado para desenvolver câncer de mama. O polimorfismo CYP1A1*2A está associado ao câncer de mama e o polimorfismo CYP1A1*2C está relacionado às mulheres com ausência de metástase à distância. Os polimorfismos dos genes GSTM1 e GSTT1 não apresentam associação com o desenvolvimento do câncer de mama.

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-04-04T11:49:33Z No. of bitstreams: 2 Dissertação - Nícolas Gustavo Matias de Oliveira - 2018.pdf: 1671663 bytes, checksum: 77209b17924ba8b6327e4442c9878a74 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-04-04T13:24:34Z (GMT) No. of bitstreams: 2 Dissertação - Nícolas Gustavo Matias de Oliveira - 2018.pdf: 1671663 bytes, checksum: 77209b17924ba8b6327e4442c9878a74 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-04-04T13:24:34Z (GMT). No. of bitstreams: 2 Dissertação - Nícolas Gustavo Matias de Oliveira - 2018.pdf: 1671663 bytes, checksum: 77209b17924ba8b6327e4442c9878a74 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-03-01 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Schizophrenia (EQZ) is a chronic mental illness that affects about 1% of the world population and is characterized by behavioral domains such as positive symptoms characterized by hallucinations and delusions and negative symptoms that involve apathy, anhedonia and social blunting. The cause of EQZ remains unknown, but it is known to be a disease whose etiology involves environmental and genetic factors. Several studies seek to identify factors that clarify the etiology of the disease. The agent that causes toxoplasmosis, Toxoplasma gondii, seems to be related to the development and progression of the disease, evidenced by studies that argue that T. gondii infection may be a triggering factor for psychosis in some individuals, since the parasite has a certain tropism by the central nervous system. Furthermore, studies have reported parasite infection as a factor related to genotoxicity, including toxoplasma infection in an animal model. Based on the foregoing, the present study aimed to evaluate the seropositivity to T. gondii (IgM or IgG) and the avidity of IgG in a group of individuals diagnosed with EQZ in the city of Goiânia and to evaluate the occurrence of genotoxicity in these, of genotoxicity with exposure to xenobiotics such as tobacco, alcohol, especially drugs, and / or T. gondii infection. Seropositivity was observed above 70% among individuals diagnosed with EQZ in relation to the controls, with avidity above 50%, indicating that individuals had contact with T. gondii at some time prior to the survey. Despite the genotoxic damage found, there was no significant difference in genotoxic damage among the infected individuals evaluated. The present study did not demonstrate that T. gondii may be a contributing factor to the occurrence of DNA damage, and the use of antipsychotic drugs, tobacco and alcohol, despite being a risk factor for genotoxicity, did not demonstrate an influence significant difference in the present study. However, it is necessary to carry out other analyzes and investigate parameters such as the time of drug use and exposure to other xenobiotics. In order to know better the life habits of individuals, it may help to provide more information about this relationship between T. gondii and EQZ. / A esquizofrenia (EQZ) é uma doença mental crônica que afeta cerca de 1% da população mundial e se caracteriza por domínios comportamentais, como sintomas positivos, caracterizados por alucinações e delírios e sintomas negativos, que envolvem apatia, anedonia e embotamento social. A causa da EQZ ainda permanece desconhecida, mas sabe-se que se trata de uma doença que cuja etiologia envolve fatores ambientais e genéticos. Vários estudos buscam identificar fatores que esclareçam a etiologia da doença. O agente causador da toxoplasmose, Toxoplasma gondii, parece ter relação com o desenvolvimento e progressão da doença, evidenciado por estudos que defendem que a infecção por T. gondii pode ser um fator desencadeante de psicoses em alguns indivíduos, já que o parasito apresenta um certo tropismo pelo sistema nervoso central. Ainda, estudos tem reportado a infecção por parasitos como fator relacionado a genotoxicidade, incluindo a infecção pelo toxoplasma em um modelo animal. Com base no exposto, o presente trabalho teve por objetivo avaliar a soropositividade para T. gondii (IgM ou IgG) e a avidez da IgG em um grupo de indivíduos diagnosticados com EQZ do município de Goiânia e avaliar a ocorrência de genotoxicidade nestes, procurando relação da genotoxicidade com a exposição a xenobióticos como tabaco, álcool, especialmente medicamentos, e/ou com a infecção pelo T. gondii. Observou- se soropositividade acima de 70% entre os indivíduos diagnosticados com EQZ em relação aos controles, com avidez acima de 50%, indicando que os indivíduos tiveram contato com o T. gondii em algum momento anterior a pesquisa. Apesar do dano genotóxico encontrado, não houve diferença significativa no dano genotóxico entre os indivíduos infectados avaliados. O presente estudo não demonstrou que T. gondii pode ser um fator contribuinte para a ocorrência de danos ao DNA, e o uso de fármacos antipsicóticos, o fumo e o álcool, apesar de serem um fator de risco para a genotoxicidade, não demonstraram uma influência significativa no presente estudo. Entretanto, há a necessidade de realizar outras análises e investigar parâmetros como o tempo de uso dos fármacos, e de exposição a outros xenobióticos, enfim, o melhor conhecimento dos hábitos de vida dos indivíduos pode ajudar a trazer mais informações acerca dessa relação entre T. gondii e a EQZ.

New medical treatments, compounds that affect human health, nutritional supplements, and other substances, are introduced to society every day. The accurate determination of the potential toxicity from these substances is of critical importance to our society. Goals of the modern toxicologist not only involve the determination of the toxic potential of new substances but also: the elucidation of mechanisms; improving existing assays; and developing new assays to study toxicity. This thesis addresses these goals in two topics fundamental to toxicology. Re-evaluating the expression of dose and susceptibility of cells in culture The exposure of cells in culture to drugs, xenobiotics, and other compounds is one of the first tools used to determine the potential for toxicity. Problems can arise when results of these experiments are translated to next-level toxicity experiments (e.g. animals and humans). I hypothesized that "dose" in cell culture can be improved by designing and reporting experiments based on dose in moles per cell. When experiments were compared on an extracellular concentration basis, a large apparent variability in toxicity was observed. However, if these same exposures were expressed as moles per cell, all experiments yielded the same toxicity. In addition to the evaluation of mole per cell, I investigated the susceptibility of various cells to 1,4-benzoquinone. I hypothesized that upon exposure to toxins that bind covalently, larger cells would require more molecules per cell of toxin versus a smaller cell to achieve identical toxicities. I found a linear correlation between cell volume(pL) and ED50 (mole per cell where 50 % cell viability is lost), supporting my hypothesis. This work could improve current cell culture protocols and allow for better and less expensive determination of toxicities. Heritability of the red blood cell storage lesion Blood transfusions are an integral part of modern medicine with 5 million people receiving blood each year in the United States. There is growing evidence that red blood cells (RBCs) stored for longer periods are less therapeutically beneficial and could even be harmful to patients. This phenomenon of diminished RBC function with increased time in storage is called the storage lesion. However, there is great variation between different donors in the severity of the storage lesion in their donated RBCs. I hypothesized that part of this variability in the RBC storage lesion is determined by heritable genetic differences. To test this hypothesis, a study using mono- and di-zygotic twins was performed to determine the heritability of adenosine triphosphate (ATP), glutathione (GSH), glutathione disulfide (GSSG) and hemolysis in stored blood. Major discoveries in this study include: GSH, GSSG, and the half-cell reduction potential (Ehc) are heritable (57 %, 51 %, and 70 %, respectively) in non-stored RBCs. In addition, ATP was found to be heritable in two different storage solutions (62 % in AS-3, 71 % in CP2D); as well as GSH, GSSG, Ehc and hemolysis (59 %, 48 %, 64 %, and 53 %, respectively). These discoveries could eventually be used to develop new genetic tests that would predict the rate of deterioration in stored blood quality on an individual basis.

Nitrochemicals are currently widely used as solvents, drugs, biocides, fuels and explosives and are consequently widely distributed in the environment. The reductive nitrite elimination from explosive compounds is catalyzed by two FMN-dependent, xenobiotic reductases (XenA or XenB). These genes for these regiospecific enzymes were cloned from Pseudomonas putida and P. fluorescens I-C respectively and isolated from the soil of a contaminated World War II munitions manufacturing plant. These enzymes enable the microbes to fulfill their nitrogen requirements from nitroglycerin by catalyzing the regiospecific, NADPH dependent, reductive denitration of nitroglycerin with differing selectivities. The two enzymes also transform a number of additional nitrocompounds in vitro, e.g. TNT and metronidazole, a leading drug in the treatment of Helicobacter pylori, a causative agent of human ulcers. Single crystals were obtained for XenA and XenB and complete X-ray diffraction datasets have been collected and analyzed to better understand these characteristics. The 1.6 Å resolution structure of XenA reveals a dimer of β/α)₈-TIM barrels, but the 2.3 Å resolution structure for XenB is a monomer. The (β/α)₈-TIM barrel protein fold is the most common fold in the PDB. However, the XenA structure exhibits a unique, C-terminal domain-swapped topology. Thus a portion of each active site is comprised of residues from the neighboring monomer. To probe the reaction cycle, crystal structures of ligand complexes and the reduced enzyme have been refined. For example, our structure of the XenA-metronidazole complex shows that ligands bind parallel to the FMN si-face. Our 1.5 Å resolution structure for reduced XenA reveals an FMN isoalloxazine ring with an angle of ~165° along the N5-N10 axis. We have also generated models of the reduced enzyme-nitroglycerin complexes by molecular dynamics. The results with both XenA and XenB reveal differences in enzyme-ligand hydrogen bonding. These differences correlate remarkably well with the regiospecific differences observed for nitrite elimination from nitroglycerin and reduction of TNT by the two enzymes.

Thesis (Ph. D.)--University of Helsinki, 2000. / Includes bibliographical references (p. 35-43). Also available in electronic format via Internet.

Malaria is a disease of major public health importance, killing approximately one million people and causing about 250 million cases of fever annually. It mostly affects children under the age of five and pregnant women in many developing countries, making it a prominent issue in international health and maternal and child health. The most aggressive form of malaria is caused by the parasite Plasmodium falciparum which is responsible for 80% of infections and 90% of deaths from malaria, and is most prevalent in sub-Saharan Africa. Public Health interventions include the implementation of prevention programs, health education, and chemotherapy. The latter has experienced multiple problems in the past years whereby resistance of the parasite to the available drugs has emerged, rendering the majority of them ineffective. Furthermore, the high cost of those drugs represents a major obstacle to their dispensation in areas of the world where the affected people are often the less fortunate. The enzyme Cytochrome b5 Reductase (cb5r) and its electron acceptor Cytochrome b5 (cb5) play a role in fatty acid elongation, cholesterol biosynthesis, and cytochrome P450-mediated detoxification of xenobiotics. Therefore, these proteins are suitable as potential novel drug targets for malaria. These two proteins have been thoroughly studied in mammals but have to be characterized in microorganisms such as fungi and parasites, including Plasmodium falciparum. It is important to note that plant cb5r has been identified as a novel herbicidal target. Considering the close phylogenetic relationship between plant cb5r and Plasmodium falciparum cb5r, we conclude that these plant inhibitors may also serve as promising candidates for a new class of antimalarial drugs against the parasite. In this project, we want to obtain the biochemical and enzymatic characterization of cb5r and cb5 in order to establish whether these two proteins represent potential novel drug targets in Plasmodium falciparum malaria. This initial work may lead to the development of novel drugs which will consequently affect the field of public health with respect to drug delivery, drug resistance, and drug chemotherapy.