Die antioxidativen Enzyme Katalase (KAT), Glutathion-Peroxidase (GPX), Glutathion-Reduktase (GR), Superoxid-Dismutase (SOD) und Glutathion-S-Transferase (GST) sind an der intrazellulären Abwehr von oxidativem Stress beteiligt. Diverse Arbeitsgruppen fanden eine Hochregulation der antioxidativen Enzyme (AOEs) nach Exposition auf Ozon. In der vorliegenden Studie sollte an einem von Schierhorn und Mitarbeitern entwickelten Kulturmodell der nasalen Mukosa des Menschen untersucht werden, ob Aktivitätsänderungen der AOEs nach Ozonexposition in vitro zu verzeichnen sind. Zu diesem Zweck wurde die nasale Mukosa von 67 Patienten, die sich wegen nasaler Atmungsbehinderung einer Conchotomie unterzogen hatten, 24 Stunden bei 37 °C und 5% CO2 kultiviert und parallel unter den selben Bedingungen einer zusätzlichen Ozonkonzentration von 120 ppb ausgesetzt. Tendenzielle Aktivitätsänderungen durch Ozon ließen sich bei der GPX (13.8 auf 17.7 mU/mg Protein, 28% Steigerung) und der SOD (8.4 auf 9.7 U/mg Protein, 15% Steigerung) feststellen. Diese Aktivitätszunahmen wiesen jedoch keine Signifikanz auf. Aktivtätsänderungen bei KAT, GR und GST durch die Ozonexposition wurden nicht gefunden. Alter der Patienten, Geschlecht und Zigarettenrauchen nahmen den Ergebnissen dieser Studie nach keinen Einfluß auf die Regulation der AOEs nach Ozonexposition. Die Deletion der Glutathion-S-Transferase M1, die bei etwa 50% der mitteleuropäischen Bevölkerung zu finden ist, veränderte die Regulation der SOD nach in vitro Ozonexposition. Die GST-defizienten Patienten dieser Studie beantworteten die Ozonexposition mit einer signifikanten Hochregulation der SOD (p / Antioxidant enzymes as catalase (CAT), glutathione peroxidase (GPX), superoxide dismutase (SOD) and glutathione S-transferase (GST) are thought the primary cellular defense mechanism against reactive oxygen species. Ozone, a highly reactive oxidant, is known to cause respiratory symptoms at ambiental doses. A number of studies have shown the mucosa of the respiratory tract to be the first target site of ozone toxicity. Other animal studies demonstrated an upregulation of mucosal antioxidant enzymes after ozone exposition. Concerning to the antioxidant defense mechanisms of the human nasal mucosa no studies are found so far. The purpose of this study was to determine if in vitro ozone exposure of human nasal mucosa results in changes in the activity of CAT, GPX, SOD, GST and glutathione reductase (GR). Nasal mucosa from 67 patients was cultivated in a specially designed in vitro organ culture and exposed to 120 ppb ozone for 24 hours. The results were compared with the histamin release which is known to be upregulated from human nasal mucosa after ozone exposition (60-200 ppb).

Made available in DSpace on 2016-08-10T10:38:42Z (GMT). No. of bitstreams: 1 KLEBER SANTIAGO FREITAS E SILVA.pdf: 1080005 bytes, checksum: 9dfcb821b9cd1574718e2442cabc533b (MD5) Previous issue date: 2013-02-18 / In healthy women, a great number of intra and extracellular controls prevent the attachment and proliferation of ectopic endometrial cells. In endometriosis, abnormalities in those controls can lead to the survival of endometrial cells and consequently their attachment to the peritoneal cavity and disease progression. Endometrial cells with genetic polymorphisms respond to local signals, and they proliferate instead of undergoing apoptosis. The products of these abnormal cells stimulate the invasion of tissues and induce an inflammatory response. The disease has a complex trait and it is related to several factors such as genetic, immunological and environmental. This study examined six polymorphisms presented by six different genes (p53; Estrogen Receptor β; Progesterone receptor; GSTM1; GSTT1; CYP1A1). We obtained the polymorphic genotype frequencies from the same 50 patients for all genes and we analyzed them using the Fisher's Exact Test or G Test. First we analyzed the genes in a group of two and subsequently in a group of three. We found significant association between polymorphisms in six pairs of genes (p53-Erβ with frequency 5.9 times higher in the experimental group; p53-GSTM1, 2.39 times higher; p53-CYP1A1 with 65.5% of the patients with the polymorphism; ERβ-PROGINS 3.0 times higher in the experimental group; GSTM1-PROGINS and GSTT1-CYP1A1 both with 31.25% of the patients with the polymorphism). Positive results were found in 15 situations when genes were analyzed in a group of three; the most significant result corresponding to the polymorphisms of the genes p53, Erβ e GSTM1 with 20% of the patients carrying these polymorphisms; PROGINS, Erβ e GSTM1 with 18% and p53, Erβ e PROGINS with 12%. The results support the idea that the presence of polymorphisms in more than one endometriosis-related gene can lead to the onset of the disease and its progression. Studies should aim at these genes in order to understand the relationship among them more clearly and the possibility of developing new diagnostic techniques based on molecular markers of these genes. / Controles intra e extracelulares impedem a implantação e a proliferação de células endometriais ectópicas nas mulheres saudáveis. Anormalidades em quaisquer desses controles levam à sobrevida dessas células, implantação e a consequente progressão da Endometriose. Células endometriais com polimorfismos genéticos respondem a sinais locais e se proliferam não sofrendo apoptose. Os produtos dessas células anormais estimulam a invasão de tecidos e induzem respostas inflamatórias. A doença é complexa e relacionada a fatores como o genético, o imunológico e o ambiental. Este trabalho analisou seis polimorfismos de seis diferentes genes (p53; Receptor β de estrógeno; Receptor de progesterona; GSTM1; GSTT1; CYP1A1). As frequências dos genótipos polimórficos foram obtidas das mesmas 50 pacientes para todos os genes e analisadas pelo Teste Exato de Fisher ou Teste G. Os genes foram analisados dois a dois e posteriormente três a três. Resultados significativos foram encontrados para seis pares de genes (p53- REβ com frequência de polimorfismo 5,9 vezes maior no grupo endometriose; p53-GSTM1 com frequência 2,39 vezes maior; p53-CYP1A1 com 65,5% das pacientes com endometriose apresentando os polimorfismos; REβ-PROGINS com frequência 3,0 vezes maior; GSTM1-PROGINS e GSTT1-CYP1A1, ambos com 31,25% das pacientes do grupo endometriose apresentando os polimorfismos). Em 15 situações quando os genes foram analisados três a três o p foi menor que 0,05. Os polimorfismos de maior frequência foram dos genes p53, REβ e GSTM1 com 20% das pacientes com endometriose apresentando esses polimorfismos; PROGINS, REβ e GSTM1 com 18% e p53, REβ e PROGINS com 12%. Esses resultados corroboram a ideia de que a presença de polimorfismos em mais de um gene relacionado à endometriose pode levar ao aparecimento e deselvolvimento da doença. Estudos devem ser direcionados a esses genes na tentativa de compreender mellhor a relação entre eles e o possível desenvolvimento de novas técnicas de diagnóstico baseada nos marcadores moleculares desses mesmos genes.

Syftet med studien var att studera effekterna av några miljöfaktorer och genetiskt predisponerande faktorer för reumatoid artrit (RA). RA är en inflammatorisk sjukdom där immunsystemet bryter ner kroppsegen vävnad. En signifikant ökad risk för RA påvisades vid exponering för spannmålsdamm och rökning. Generna som studerades var GSTM1, GSTT1 och Fc-gamma-RII. Generna i sig var ingen predisponerande faktor för RA, men rökning visade sig vara en signifikant högre riskfaktor för de som hade en deletion av GSTM1 eller Fc-gamma-RII genen samt för de som inte hade deletion av GSTT1 genen. Exponering för spannmålsdamm visade sig vara en signifikant större riskfaktor för RA för de som inte hade deletion av GSTM1, GSTT1 eller Fc-gamma-RII genen. Vidare visade individer som inte hade en deletion av Fc-gamma-RII genen en ökad risk för RA vid exponering för stendamm. / The aim of this study was to evaluate the effects of some environmental and genetic predisposing factors of Rheumatoid Arthritis (RA). RA is an inflammatory disease where the immune system decomposes body tissue. A significant increased risk of RA was shown when exposed to grain and smoking. The genes that were studied were GSTM1, GSTT1 and Fcgamma- RII. The genes themselves were no predisposing factors of RA, but smoking showed to be a significant greater riskfactor for RA for individuals having a deletion of the GSTM1 or the Fc-gamma-RII gene, or not having a deletion of the GSTT1 gene. Exposure to grain showed to be a significant greater riskfactor for RA for individuals not having a deletion of the GSTM1, GSTT1 or the Fc-gamma-RII gene. Also individuals not having a deletion of the Fc-gamma-RII gene showed an increased risk for RA when exposed to stonedust.

<p>Glaucoma is one of the leading causes of visual impairment in the world. In glaucoma, the patient’s peripheral vision is lost due to progressive and irreversible deterioration of the retinal ganglion cells and atrophy of the optic nerve. The effect on the visual field is gradual and painless, and the progression so slow, that the patient may not notice until a substantial part of the visual field is lost. If left untreated, glaucoma can lead to blindness.</p><p>In this thesis, genes associated to glaucoma have been analysed in Swedish patients with primary open angle and exfoliative glaucoma. The genes studied were <i>MYOC</i>, <i>oculomedin,</i> <i>GSTM1</i> and <i>OPTN</i>.</p><p>The coding sequence of <i>MYOC</i> was analysed and mutations were found in 1% of the primary open angle glaucoma patients. Additionally, a predisposing variant was found in 1% of the patients as well as in 0.5% of the controls. No disease-associated variation was found in the exfoliative glaucoma cases. Mutations were also found in two families affected by glaucoma. The coding sequence of <i>oculomedin</i> was analysed, but none of the variants found were classified as disease causing in either patient group. <i>GSTM1</i> was analysed for its presence in the patients. No association could be found for either hetero- or homozygous deletions. The coding sequence and haplotype distribution of <i>OPTN</i> was analysed. None of the variants found were classified as disease causing and none of the haplotypes were associated to the disease in either patient group.</p><p>There are just a few per cent of the Swedish primary open angle glaucoma patients with genetic variation associated to disease, in the genes analysed in this study. No association to exfoliative glaucoma was found. This indicates heterogeneity in the genetics of glaucoma when different subtypes and different populations are compared. Likely, there are genes still to be identified.</p>

Glaucoma is one of the leading causes of visual impairment in the world. In glaucoma, the patient’s peripheral vision is lost due to progressive and irreversible deterioration of the retinal ganglion cells and atrophy of the optic nerve. The effect on the visual field is gradual and painless, and the progression so slow, that the patient may not notice until a substantial part of the visual field is lost. If left untreated, glaucoma can lead to blindness. In this thesis, genes associated to glaucoma have been analysed in Swedish patients with primary open angle and exfoliative glaucoma. The genes studied were MYOC, oculomedin, GSTM1 and OPTN. The coding sequence of MYOC was analysed and mutations were found in 1% of the primary open angle glaucoma patients. Additionally, a predisposing variant was found in 1% of the patients as well as in 0.5% of the controls. No disease-associated variation was found in the exfoliative glaucoma cases. Mutations were also found in two families affected by glaucoma. The coding sequence of oculomedin was analysed, but none of the variants found were classified as disease causing in either patient group. GSTM1 was analysed for its presence in the patients. No association could be found for either hetero- or homozygous deletions. The coding sequence and haplotype distribution of OPTN was analysed. None of the variants found were classified as disease causing and none of the haplotypes were associated to the disease in either patient group. There are just a few per cent of the Swedish primary open angle glaucoma patients with genetic variation associated to disease, in the genes analysed in this study. No association to exfoliative glaucoma was found. This indicates heterogeneity in the genetics of glaucoma when different subtypes and different populations are compared. Likely, there are genes still to be identified.

The aetiology of Parkinson's disease (PD) is still unclear. Research findings suggest that both environmental and genetic factors may contribute to its development. The interactions between genes and the environment might exist and play a key role. Cigarette smoking was found to be one of the few factors exhibiting a protective effect. If chemical compounds found in cigarette smoke influence PD risk, the difference in the ability of certain individuals in metabolising these substances might alter their susceptibility to the risk of developing PD. Many metabolic enzyme genes exhibit polymorphic traits with alteration of gene function. These might be associated with an altered susceptibility of individuals to PD. Few studies have examined the hypothesis that metabolic enzyme gene polymorphisms might modulate the effect of smoking on PD risk. However, it is crucial to consider these potential interactions when we try to elucidate the aetiology of PD. Even if each factor only contributes a slight variation and influences a small portion of the whole population, non-linear and unpredictable interactions may account for a high proportion of the aetiological fraction. Previous studies have not been strictly designed to examine the interactions between smoking and metabolic enzyme genetic polymorphisms. These studies have not been able to elucidate the extent of the interaction. Therefore, this PhD project attempted to examine whether genetic factors, operating in the phase one and phase two metabolic pathways, interact with smoking to influence the development of PD. This is the first genetic epidemiological study of PD specifically addressing this issue. The research aids in further understanding the aetiology of PD and may be useful for identifying people at higher risk. A case-only design was chosen for this project for two reasons: first, PD is a relatively rare disease and the case-only design is much more efficient at detecting gene-environment interactions; second, the PD cases for the project were recruited over the past few years and represent a prevalence series, for which an appropriate comparison group for the cases is difficult to identify and recruit. In a case-only study, only cases are used to investigate the multiplicative effects of the exposures and susceptible genotypes of interest, while non-case subjects (traditionally controls) are solely used to test the independence between the exposure and the susceptible genotype. Therefore, this approach avoids the challenges of control selection, a major limitation inherent in the case-control approach. This thesis comprised of three independent studies: the first study investigated the interactions between genetic polymorphisms of GSTM1, P1, T1 and Z1 and smoking in PD; the second study examined the interactions between genetic polymorphisms of CYP2E1 and smoking in PD; and the third study examined the interactions between genetic polymorphisms of CYP2D6 and smoking in PD. The first two studies recruited 400 white Caucasian PD cases from both hospital wards and private neurology clinics (230 men and 170 women). The third study further included 142 white Caucasian PD cases newly recruited from the same sources (542 in total, 321 men, and 221 women). The mean age of cases was 67 years with the average onset age at 60 years. GSTM1, GSTP1, GSTT1, GSTZ1 AND CYP2E1 genotyping processes were performed using protocols previously published with minor modification, whereas CYP2D6 genotyping methods were mainly developed by me with assistance from associate supervisor Dr. George Mellick. Reliability and validity of the PCR and RFLP methods were assessed through re-conducting the genotype assays using at least a 10% sample of our DNA samples. The results for all re-assessments were 100% concordant. Crude bivariate analyses were adjusted for potential confounding effects of the variables, including age at onset, gender, family history of PD and pesticide exposures. Among our unaffected, aged subjects (mean age: 63.9 years, sd: 11.4 years), the genotype frequencies at each locus were similar to those reported in other Caucasian populations. The first study showed that the proportion of carriers of the GSTP1-114Val allele (mutant) increased with increasing smoking dose from 0 to > 30 pack-years. Homozygotes of the 114Ala allele (wild-type) decreased with increasing smoking dose (trend test: p=0.02). This trend existed both in male and female cases. This dose-effect relationship was most significant in the group of cases with late-onset PD (i.e., age at onset > 55 years) with the ORicase-only values of 1.88 (95%CI: 0.65-5.48) and 2.63 (95%CI: 1.07-6.49) for > 0-10 and > 10 pack-years, respectively. No similar trend was found among our unaffected, aged subjects (p=0.42). Haplotype analyses revealed significant differences for GSTP1 haplotypes between smoking and non-smoking PD cases (ORicase-only for *C haplotype=2.00 (95%CI: 1.11-3.60), p=0.03). In this case, smoking-exposed PD cases were more likely to posses the *C haplotype defined by A to G base-pair transition at nucleotide +313 and C to T base-pair transition at nucleotide +341 (at amino acid level, valine at both positions 105 and 114). The second study found no difference in CYP2E1 genotype frequencies between PD cases who ever smoked compared to those who never smoked (odds ratio for interaction (ORi) = 1.00 (95% CI: 0.39-2.51, p=0.99)). No CYP2E1 gene-smoking interactions were detected in relation to age at onset of PD. The third study found that among cases without regular pesticide exposures, CYP2D6 PMs who smoked more than 5 pack-years had a later mean age at disease onset (68.6 years) than those with extensive metaboliser phenotypes (EMs) (61.1 years, p=0.02) and non-smokers (60.5 years, p=0.01). Analysis of aged subjects without PD confirmed that neither smoking status nor CYP2D6 PM status was associated with age itself. Our data suggest: 1. smoking exposure is independent of GSTM1, P1, T1, Z1 and CYP2E1 genotypes; 2. smoking may be, to some extent, associated with CYP2D6 genotypes; 3. there are no multiplicative interactive effects linking smoking and GSTM1, T1, Z1 or CYP2E1 genotypes with the risk for PD; 4. there is a multiplicative interactive effect between smoking and GSTP1 haplotype - particularly for genotypes carrying the 114Val allele; and 5. there is a multiplicative interactive effect between smoking and CYP2D6 PMs - particularly for people who ever smoked cigarettes more than 5 pack-years. In general, this thesis provides a model for exploring the gene-smoking interactions in PD. Further studies need to consider the recruitment of a large number of population-based and randomly-selected samples and to pay more attention to measurement of environmental exposures. Further studies also need to examine simultaneously the impact of smoking, pesticide exposures and other potential risk factors on PD. These studies will build evidence for interactions contributing to this common neurological movement disorder.

Made available in DSpace on 2014-06-11T19:32:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-08-28Bitstream added on 2014-06-13T20:03:26Z : No. of bitstreams: 1 ondei_ls_dr_sjrp_parcial.pdf: 292639 bytes, checksum: 9c76afbfba65412651952af8454cb31d (MD5) Bitstreams deleted on 2015-01-16T10:37:50Z: ondei_ls_dr_sjrp_parcial.pdf,Bitstream added on 2015-01-16T10:38:34Z : No. of bitstreams: 1 000603676.pdf: 889558 bytes, checksum: df390d92da0411515e31635f99e1d76d (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Ministério da Saúde / Na talassemia beta, o acúmulo das cadeias alfa livres, bem como a liberação do grupo heme e do ferro durante o processo hemolítico, ocasionam aumento de danos oxidativos que podem resultar em lipoperoxidação de membranas celulares, desnaturação de proteínas e oxidação da hemoglobina. Na deficiência de glicose- 6-fosfato desidrogenase (G6PD), esse aumento é decorrente da diminuição da produção de nicotinamida adenina dinucleotídeo fosfato reduzido (NADPH) que pode resultar em hemólise intravascular. Diante da possibilidade de estresse oxidativo nos portadores de beta talassemia heterozigota e nos indivíduos com deficiência de G6PD, neste trabalho avaliou-se a expressão fenotípica das afecções genéticas por meio da identificação das mutações e análise de marcadores para estresse oxidativo. Para o estabelecimento dos grupos controle e com deficiência de G6PD foram avaliadas 544 amostras de sangue periférico de indivíduos da região Noroeste do Estado de São Paulo, sendo 426 doadores de sangue e 118 indivíduos de uma instituição de ensino superior. Para a composição do grupo com talassemia beta heterozigota foram avaliadas 46 amostras de sangue de indivíduos com diagnóstico clínico de talassemia beta da cidade de São Carlos/SP. Foram realizados métodos de triagem e confirmatórios para a identificação da talassemia beta heterozigota e da deficiência de G6PD, e dosagens bioquímicas para quantificação das espécies reativas ao ácido tiobarbitúrico (TBARS), utilizado como marcador de estresse oxidativo, e para a determinação da capacidade antioxidante em equivalência ao Trolox (TEAC). Os polimorfismos da glutationa S-transferase (GST) GSTM1 e GSTT1 foram avaliados por PCR multiplex o de GSTP1 por PCR/RFLP. No grupo com talassemia beta heterozigota foram encontradas 18 (39%) amostras com a mutação CD39; 22 (48%) com a mutação... / In beta thalassemia, the excess of unpaired alpha chains, as well as the heme group and iron released during the hemolytic process increase the oxidative damage. In G6PD deficiency, this increase is caused by a reduced production of NADPH that results in an intravascular hemolysis. Thus, facing the oxidative stress possibility in beta thalassemia carriers and G6PD deficiency individuals, it was aimed to evaluate the fenotypic expression of this genetic disorders through the mutation identification, as well as the oxidative stress marker analysis. We used 544 peripheral blood samples of individuals from São Paulo’s northwestern to control group and to G6PD deficiency group establishment. For beta thalassemia heterozygote group were evaluated 48 blood samples of São Carlos/SP city. Tests were carried out aiming the screening and confirmation of beta thalassemia and G6PD deficiency, as well as the analysis of lipid peroxidation products measured as thiobarbituric acid reactive species (TBARS) and Trolox equivalent antioxidant capacity (TEAC). Were determined the frequencies of GSTM1, GSTT1 and GSTP1 polymorphisms. The analysis with beta thalassemia carriers allowed to establish in the study group a frequency of 39% for CD39 mutation, 48% for IVS-I-110 mutation and 2% for IVS-I-6 mutation. For G6PD deficiency was founded a frequency of 3.86%. The beta thalassemic group evaluation showed an increase of TBARS and TEAC values, when compared to the control group. There was a tendency to increase lipid peroxidation in beta0 CD39 mutants compared to beta+ IVS-I-110 mutants, because there is more free chains amount in beta0 thalassemia than beta+ thalassemia. In the G6PD deficiency analysis was found a lower G6PD activity in men than in women, but there was no interference of gender in the TBARS and TEAC assays results. The comparison between the control group and the G6PD deficiency group... (Complete abstract click electronic access below)

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2014-10-30T18:14:19Z No. of bitstreams: 2 Dissertação - Denise da Silva Pinheiro.pdf: 1388647 bytes, checksum: e26020f5d13976c2a4920327b71bc5c4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-10-31T09:40:46Z (GMT) No. of bitstreams: 2 Dissertação - Denise da Silva Pinheiro.pdf: 1388647 bytes, checksum: e26020f5d13976c2a4920327b71bc5c4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-10-31T09:40:46Z (GMT). No. of bitstreams: 2 Dissertação - Denise da Silva Pinheiro.pdf: 1388647 bytes, checksum: e26020f5d13976c2a4920327b71bc5c4 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-08-30 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / In the pathogenesis of type 2 diabetes mellitus (DM2), it is observed that the increased oxidative stress may contribute to decreased insulin production and destruction of pancreatic β cells. Individuals who have no activity of GSTM1 and GSTT1 isoforms may have an increased susceptibility to damage caused by reactive species to pancreatic β cells, since these cells express low levels of antioxidant enzymes. This case-control study aimed to analyze the genotypic profiles of the deletion polymorphism of GSTM1 and GSTT1 genes by molecular assays (conventional PCR and qPCR) to associate these polymorphisms with DM2 risk, considering that studies with this approach have not been conducted in Brazil. Data of clinical, laboratorial and demographic variables of 120 patients and 147 controls were obtained through interviews and information from medical charts (patients) or results of recent clinical and laboratory exams (controls). It was found that diabetic patients had a higher frequency of GSTT1-null genotype (29.2%) than non-diabetic subjects (12.2%), and those with the risk genotype have an increased predisposition to diabetes from approximately 3.2 times (p = 0.0004). However, there was no association of GSTM1-null with DM2 susceptibility. The analysis of the influence of GSTT1 deletion on clinical and biochemical changes in the case group showed that the risk genotype may contribute to the development of DM2 complications related to dyslipidemia, due to the association of GSTT1-null with significantly higher levels of triglycerides (p = 0.0242) and VLDL-cholesterol levels (p = 0.0252) compared to patients without the risk genotype. Additionally, GSTM1-null was associated with elevated levels of fasting glucose, glycated hemoglobin and blood pressure. We emphasized a necessity for applying log-linear analysis in order to explore an interaction between these polymorphisms properly. These results suggest that GSTT1 polymorphism may play an important role in the pathogenesis of DM2 in Brazilian population. Then, this gene could be added to a set of genetic markers to identify individuals at increased risk for developing DM2. Although there was no association of GSTM1 deletion polymorphism with susceptibility to DM2, it was verified the influence of this polymorphism on important clinical parameters related to glycemia and blood pressure levels. This finding suggests that GSTM1-null, GSTT1-null as well, may contribute to the clinical course of diabetic patients. / Na patogênese do diabetes mellitus tipo 2 (DM2), observa-se que o estresse oxidativo aumentado pode contribuir na diminuição da produção de insulina e destruição das células β pancreáticas. Indivíduos que apresentam ausência de atividade das isoformas GSTM1 e GSTT1 podem apresentar uma susceptibilidade aumentada aos danos causados por espécies reativas às células β pancreáticas, uma vez que estas células expressam baixos níveis de enzimas antioxidantes. O presente estudo caso-controle visou analisar os perfis genotípicos do polimorfismo de deleção dos genes GSTM1 e GSTT1 por ensaios moleculares (PCR convencional e qPCR) para associar tais polimorfismos com o risco ao DM2, considerando que ainda não foram realizados estudos com este enfoque no Brasil. Dados clínicolaboratoriais e demográficos de 120 pacientes e 147 controles foram obtidos por meio de entrevistas e consulta a prontuários (pacientes) e a resultados de exames recentes (controles). Foi verificado que os pacientes diabéticos apresentaram uma frequência mais elevada de genótipo GSTT1-nulo (29,2%) do que indivíduos não-diabéticos (12,2%), e que aqueles que apresentam o genótipo de risco possuem uma predisposição aumentada a diabetes de aproximadamente 3,2 vezes (p = 0,0004). No entanto, não houve associação de GSTM1-nulo com a susceptibilidade ao DM2. A análise da influência da deleção de GSTT1 sobre alterações bioquímicas e clínicas no grupo caso demonstrou que o genótipo de risco pode contribuir para o desenvolvimento de complicações do DM2 relacionadas à dislipidemia, em função da associação de GSTT1-nulo com níveis significativamente mais elevados de triglicérides (p = 0,0242) e VLDL-colesterol (p = 0,0252) em comparação aos pacientes sem o genótipo de risco. Adicionalmente, GSTM1-nulo teve associação com níveis elevados de glicemia de jejum, hemoglobina glicada e pressão sanguínea. Foi enfatizada a necessidade de aplicação da análise log-linear para investigar a interação entre os polimorfismos apropriadamente. Os resultados obtidos sugerem que o polimorfismo de deleção de GSTT1 pode desempenhar um importante papel na patogênese do DM2 na população brasileira. Portanto, este gene poderia ser adicionado a um painel de marcadores genéticos para identificação de indivíduos com alto risco ao desenvolvimento do DM2. Embora não tenha ocorrido associação do polimorfismo de deleção de GSTM1 com a susceptibilidade ao DM2, foi verificada a influência deste polimorfismo sobre importantes parâmetros relacionados ao controle glicêmico e pressórico. Estes achados sugerem que GSTM1-nulo, assim como GSTT1-nulo, podem contribuir para a evolução clínica dos pacientes diabéticos.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-07-13T20:37:59Z No. of bitstreams: 2 Dissertação - Rayane Mendes de Lima - 2016.pdf: 1517541 bytes, checksum: 21e56d88c3ecbe596bbba9be9f3ea87e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-07-14T13:04:34Z (GMT) No. of bitstreams: 2 Dissertação - Rayane Mendes de Lima - 2016.pdf: 1517541 bytes, checksum: 21e56d88c3ecbe596bbba9be9f3ea87e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-07-14T13:04:34Z (GMT). No. of bitstreams: 2 Dissertação - Rayane Mendes de Lima - 2016.pdf: 1517541 bytes, checksum: 21e56d88c3ecbe596bbba9be9f3ea87e (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-03-02 / Diabetic nephropathy is the leading cause of end-stage renal disease (ESRD) in developed countries and in the literature shows as oxidative stress possibly contributes to the development of the diseases. Glutathione S-transferases (GSTs) are a family of multifunctional enzymes that play an important role in the cellular detoxification and excretion of numerous substances and can also work as one of the antioxidants. The genetic polymorphism of deletion in GSTT1 and GSTM1 gene, when homozygous, show lack of activity of these isoforms, known as null genotype. Looking for a possible relationship between diabetic nephropathy and polymorphisms mentioned above, this study was made for the case-control and genotyping using real-time PCR (qPCR) and melting curve. Clinical and laboratorial data of 65 patients (diagnosed with diabetic nephropathy and were on hemodialysis) and 90 controls were collected through interviews and consultation with medical records (patients) and the results of recent surveys (controls). It was found that in the group if there is a risk associated with deletion polymorphism, where the GSTT1-null genotype (p = 0,0230) causes an increased risk of about 2,9 times in developing the disease (diabetic nephropathy) compared to carriers of the genotype GSTT1-present. There was no association of GSTM1 (p = 0.3860) with susceptibility to disease in this population. Analysis of the influence of the deletion of GSTT1 and GSTM1 about the biochemical and clinical changes in the group case did not result in a significant association in any of the clinical variables analyzed. These results suggest that the GSTM1 deletion polymorphism may be associated with risk of developing the disease, but not with the biochemical changes that were analyzed. Further studies may clarify the relationship of this polymorphism with diabetic nephropathy and help in the treatment of this disease. / A nefropatia diabética é a principal causa de Estágio Final de Doença Renal (ESRD) em países desenvolvidos e evidências tem apontado o estresse oxidativo como unificador de várias vias de dano celulares em condições de hiperglicemia, que resultaria no desenvolvimento e complicações da doença. As glutationa-S-transferases (GSTs) são uma família de enzimas multifuncionais que desempenham um papel importante na desintoxicação celular e eliminação de numerosas substâncias e também pode funcionar como um dos antioxidantes. O polimorfismo genético de deleção nos genes GSTT1 e GSTM1, quando em homozigose, apresentam ausência de atividade dessas isoformas, conhecido como genótipo nulo. Procurando estabelecer uma possível relação entre nefropatia diabética e os polimorfismos acima mencionados, foi feito um estudo por caso-controle e a genotipagem por meio de PCR em tempo real (qPCR) e curva de melting. Dados clínico-laboratoriais de 65 pacientes (diagnosticados com nefropatia diabética e que estavam em hemodiálise) e 90 controles foram coletados, por meio de entrevistas e consulta a prontuários (pacientes) e a resultados de exames recentes (controles). Foi verificado que no grupo caso existe um risco associado ao polimorfismo de deleção, onde o genótipo GSTT1-nulo (p=0,0230) provoca um risco aumentado de aproximadamente 2,9 vezes em desenvolver a doença (nefropatia diabética) em relação aos portadores do genótipo GSTT1-presente. Não houve associação de GSTM1 (p=0,3860) com a susceptibilidade a doença na população estudada. A análise da influência da deleção de GSTT1 e GSTM1 sobre as alterações bioquímicas e clínicas no grupo caso não resultou em uma associação significativa de nenhuma das variáveis clínicas analisadas. Estes resultados sugerem que o polimorfismo de deleção de GSTT1 pode estar associado ao risco de desenvolvimento da doença, mas não com as alterações bioquímicas que foram analisadas. Novos estudos poderão esclarecer a relação desse polimorfismo com a nefropatia diabética e auxiliar no tratamento desta doença.

Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2016-09-12T20:26:04Z No. of bitstreams: 2 Dissertação - Raimundo Lima da Silva Junior - 2008.pdf: 5070455 bytes, checksum: 3393ae35ddba4af1c14d43ce110ef279 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-09-15T12:45:04Z (GMT) No. of bitstreams: 2 Dissertação - Raimundo Lima da Silva Junior - 2008.pdf: 5070455 bytes, checksum: 3393ae35ddba4af1c14d43ce110ef279 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2016-09-15T12:45:04Z (GMT). No. of bitstreams: 2 Dissertação - Raimundo Lima da Silva Junior - 2008.pdf: 5070455 bytes, checksum: 3393ae35ddba4af1c14d43ce110ef279 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2008-05-30 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The larynx is a tubular organ of the aerodigestive tract whose main function is the production of voice, in addition to tasks related to the protection of the lower airways and swallowing food. The squamous cell carcinoma presents itself as a multifactorial disease and epigenetic being influenced by environmental factors, behavioural and relate to the individual. Several polymorphic genes that encode enzymes involved in the biotransformation of the carcinogens have been associated to the development of cancer, especially in squamous cell carcinoma of head and neck. The CYP1A1m, GSTM1 and GSTT1 genes which catalyzes the oxidation of PAHs in phenolic and epoxides products. The purpose of this study was to evaluate the epidemiological data of patients diagnosed with squamous cell carcinoma of the larynx from the Cancer Registry of Population basis of Goiânia and from Association of Cancer Combat in Goiânia, analyzing data from 63 medical records followed in a period of 6 year; establish the allelic profiles of the gene CYP1A1m, GSTM1 and GSTT1 in casecontrol groups investigating the degrees of suscetibility of patients diagnosed with squamous cell carcinoma (SCC) of the larynx, according to smoking and alcoholism habits. In total 112 samples were evaluated, 63 from patients with SCC of the larynx and 49 patients in the control group. The CEC of the larynx was about 3,5 times more frequent in men than in women. The rate of survival on general was approximately 66% to 6 years. The smoking and ethilic habits associated have a significantly lower survival. No significant differences were found in the staging analysis and treatment of patients. Were found allelic frequencies of CYP1A1m1 of 52,4% (34/62) and 63,3% (31/49) for the wild allele T and 44,4% (28/62) and 34,7% (18/49) for the mutant allele C. There were no statistically significant associations between allelic variants and genotypic CYP1A1m1 with squamous cell carcinoma of the larynx. Positive genotypic frequencies were observed for the genes GSTM1 and GSTT1 of 41.3% (26/63) and 54.0% (34/63) to the patients with laryngeal cancer, and 49,0% (24/49) and 51,0% (25/49) for the control group, respectively. For the assessment of genotype GSTM1 (null) and GSTT1 (null), the patients if the case group had 58,7% (37/63) and 46,0% (29/63) of genotypes for GSTM1 null and GSTT1, while the patients in the control group showed 51,0% (25/49) and 40,8% (20/49). There was no statistically significance in associations of genotypic variants GSTM1 (null) and GSTT1 (null) with the larynx squamous cell carcinoma. In this context, an experimental study was accomplished in the intention of evaluating the function of the genes Cyp and Gst in the metabolism of the Cyclophosphamide. This study addressed the Cyclophosphamide toxicological evaluation and analysis of the polymorphism of Cyp2a29 and Gstp1 BALB/c genes in mice, correlating them with the toxicity of drugs through the assessment of organs commonly affected. It was determinated the DL50 of the 474mg/kg. Subsequently, another 40 mice were used on the assessment of the genetic polymorphism of genes Cyp2a29 and Gstp1. As for toxicity, cyclophosphamide caused reduction in body weight at different doses and kidneys, spleen, liver, heart and lungs. The animals that received a dose of 500mg/Kg died in 24 hours, three animals with a dose of 250mg/Kg died the end of one week. The evaluation of toxicity showed changes of varying degrees of congestion observed in the spleen, liver and heart. There were not found statistically significant differences between the frequencies of allelic and genotypic to Cyp2a29 and Gstp1 in groups of animals studied. This observation should be probably the characteristic line BALB/c are isogenic. Studies that make the use se of tests of genetic suscetibility, epidemic evaluation, combined to the evaluation anatomopathological they allow the study of the interaction genetic atmosphere and a possible standardization of various chemotherapy protocols for treatment. / A laringe é um órgão tubular do trato aerodigestivo cuja principal função é a produção da voz, além de funções relacionadas com a proteção das vias aéreas inferiores e deglutição alimentar. O carcinoma de células escamosas (CEC) de laringe apresenta-se como uma doença multifatorial e epigenética sendo influenciada por fatores ambientais, comportamentais e inerentes ao indivíduo. Vários genes polimórficos que codificam enzimas envolvidas na biotransformação de carcinógenos têm sido associados ao desenvolvimento de câncer, sobretudo no carcinoma espinocelular de cabeça e pescoço. Os genes CYP1A1m1, GSTM1 e GSTT1 são fundamentais no processo de detoxificação de Poli - Hidrocarbonetos Aromáticos (PHA) e de diversas moléculas endógenas, facilitando o seu metabolismo e excreção. Os objetivos do presente estudo foram de avaliar os dados epidemiológicos de pacientes diagnosticados com CEC da laringe do Registro de Câncer de Base Populacional de Goiânia (RCBP) da Associação de Combate ao Câncer em Goiás (ACCG), analisando os dados de 63 prontuários acompanhados num período de 6 anos; estabelecer os perfis alélicos dos genes CYP1A1m1, GSTM1 e GSTT1 em grupos caso-controle investigando os graus de suscetibilidade dos pacientes diagnosticados com CEC de laringe, segundo hábitos tabagistas e etilistas. Ao total foram avaliadas 112 amostras, sendo 63 provenientes de pacientes com CEC da laringe e 49 de pacientes do grupo controle. O CEC da laringe se mostrou cerca de 3,5 vezes mais freqüente em homens que em mulheres. A taxa de sobrevida relativa geral foi de aproximadamente 66% para 6 anos. Os hábitos tabagista e etilista concomitantes apresentam uma sobrevida consideravelmente menor. Foram encontradas freqüências alélicas para CYP1A1m1 de 52,4% (34/62) e 63,3% (31/49) para o alelo selvagem T e 44,4% (28/62) e 34,7% (18/49) para o alelo mutante C. Não foram encontradas associações estatisticamente significativas das variantes alélicas e genotípicas CYP1A1m1 para o CEC da laringe. Quanto aos genes GSTM1 e GSTT1 foram observadas freqüências genotípicas positivas para os genes GSTM1 e GSTT1 de 41,3% (26/63) e 54,0% (34/63) para os pacientes com carcinomas laríngeos, e de 49,0% (24/49) e 51,0% (25/49) para o grupo controle, respectivamente. Para a avaliação do genótipo GSTM1 (nulo) e GSTT1 (nulo), os pacientes do grupo caso apresentaram 58,7% (37/63) e 46,0% (29/63) de genótipos nulos para GSTM1 e GSTT1, enquanto que os pacientes do grupo controle apresentaram 51,0% (25/49) e 40,8% (20/49). Não foram encontradas associações estatisticamente significativas das variantes genotípicas GSTM1 (nulo) e GSTT1 (nulo) para o CEC da laringe. Neste contexto, foi realizado um estudo experimental no intuito de avaliar o papel dos genes Cyp e Gst no metabolismo da ciclofosfamida. Tal estudo abordou avaliações toxicológicas da Ciclofosfamida e a análise do polimorfismo dos genes Cyp2a29 e Gstp1 em camundongos BALB/c e correlacionando-os com a toxicidade da droga através da avaliação dos órgãos comumente afetados. A DL50 da droga observada em 40 camundongos foi de 474mg/Kg. Posteriormente, outros 40 camundongos foram utilizados na avaliação do polimorfismo genético dos genes Cyp2a29 e Gstp1. Quanto à toxicidade, a ciclofosfamida ocasionou redução da massa corporal nos rins, baço, fígado, coração e pulmões. A avaliação da toxicidade demonstrou alterações de diferentes graus de congestão observados no baço, fígado e coração. Não foram encontradas diferenças estatisticamente significativas entre as freqüências alélicas e genotípicas de Cyp2a29 e Gstp1 nos grupos dos animais estudados. Estudos que preconizem a utilização de testes toxicológicos combinados à avaliação anatomopatológica e investigação da variabilidade genética permitem a padronização de protocolos de tratamento de diversos quimioterápicos.