Submitted by admin tede (tede@pucgoias.edu.br) on 2016-11-30T16:52:02Z No. of bitstreams: 1 FÁBIO LEMOS CAMPEDELLI.pdf: 8948383 bytes, checksum: 229016125b12d4f344bb1c157148ddad (MD5) / Made available in DSpace on 2016-11-30T16:52:02Z (GMT). No. of bitstreams: 1 FÁBIO LEMOS CAMPEDELLI.pdf: 8948383 bytes, checksum: 229016125b12d4f344bb1c157148ddad (MD5) Previous issue date: 2016-08-12 / INTRODUCTION: Atherosclerotic disease (AD) with its cardiovascular complications is responsible for 17.5 million deaths a year, according to the World Health Organization (WHO). There is consensus that atherosclerosis involves multiple pathogenic processes initiated by endothelial dysfunction, with inflammation and vascular proliferation determining alterations in the matrix, with consequent formation of the atheromatous plaque and its clinical implications. Risk factors such as hypertension, diabetes mellitus, dyslipidemia and smoking are widely known. Currently genotyping, which is not directly related to these factors, is not accepted to estimate the risk of cardiovascular diseases, but strong evidence indicates several polymorphic genes as factors of risk and progression leading to complications of the disease. Among the genes involved, eNOS (endothelial nitric oxide synthase gene), which is responsible for the production of endothelial nitric oxide (NO, an important arterial vasodilator), when presented in polymorphic variation can determine production malfunction and predisposition to AD. OBJECTIVES: To analyze the G894T polymorphism of eNOS gene in groups of individuals diagnosed with atherosclerosis and in the control group. MATERIALS AND METHODS: 200 blood samples were collected from patients previously diagnosed with DA and 100 from the control group. The genotyping analysis for polymorphism of eNOS gene was determined by PCR. RESULTS: After analysis of polymorphism between the DA and control groups and association with variables such as gender, relation with smoking, smoking history and alcohol consumption, statistical difference were found in the distribution of the case and control groups (p = 0.0378) and in non-smoking patients (p = 0.0263). In the other associations no statistically significant difference was found. CONCLUSION: In the population studied, the frequency of the heterozygous genotype (GT) was much higher than in the other popualations (GG and TT) in both groups (case and control). The GG genotype showed greater susceptibility to AD. The association of GG genotype in nonsmokers also showed greater susceptibility. Gender, alcohol consumption, smoking and smoking history did not influence AD. / Campedelli FL. Polimorfismo do gene eNOS G894T (Glu298Asp) em pacientes sintomáticos para aterosclerose [dissertação]. Goiânia: Pontifícia Universidade Católica, PUC-GO; 2016. INTRODUÇÃO: A doença aterosclerótica (DA) com suas complicações cardiovasculares é responsável por 17,5 milhões de mortes por ano segundo a Organização Mundial de Saúde (OMS). É consenso que a aterosclerose envolve múltiplos processos patogênicos que se iniciam pela disfunção endotelial, com inflamação e proliferação vascular determinando alterações da matriz com consequente formação da placa ateromatosa e suas repercussões clínicas. Fatores de risco como a hipertensão arterial, diabetes mellitus, dislipidemias e tabagismo são amplamente conhecidos. Atualmente a genotipagem, não relacionada diretamente a estes fatores, não é aceita para estimativa de risco das doenças cardiovasculares, porém fortes evidências relacionam diversos genes polimórficos, como fator de risco e evolução para complicações da doença. Dentre os genes envolvidos o eNOS (gene da síntese de óxido nítrico endotelial), responsável pela produção de Óxido nítrico (NO), endotelial (importante vasodilatador arterial), quando se apresenta em variação polimórfica pode determinar mal funcionamento da produção e predispor a DA. OBJETIVOS: Analisar o polimorfismo G894T do gene eNOS nos grupos de indivíduos com diagnóstico de aterosclerose e no grupo controle. MATERIAIS E MÉTODOS: Foram coletados amostras de sangue periférico de 200 pacientes com DA previamente diagnosticados e 100 amostras de grupo controle. A análise de genotipagem para o polimorfismo do gene eNOS foi determinada por PCR. RESULTADOS: Após análise do polimorfismo entre os grupos com DA e grupo controle, e associação com as variáveis gênero, relação com hábito de fumar, carga tabágica e consumo de bebida alcóolica, foram encontrados diferença estatísticas na distribuição dos grupos caso e controle (p=0,0378) e nos pacientes não tabagistas (p=0,0263). Nas demais associações não houve diferença estatisticamente significativa. CONCLUSÃO: Na população estudada evidenciou a frequência do genótipo heterozigoto (GT) muito superior aos demais (GG e TT) em ambos os grupos (caso e controle). O genótipo GG demonstrou maior suscetibilidade à DA. A associação do genótipo GG em não tabagista também apresentou maior suscetibilidade. O gênero, o etilismo, o hábito de fumar e carga tabágica não influenciaram na DA.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-01-13T12:38:13Z No. of bitstreams: 2 Tese - Kênia Alves Pereira Lacerda - 2016.pdf: 1930787 bytes, checksum: d2d35012cd70eaa30ae8e03b674c7055 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-01-16T11:01:34Z (GMT) No. of bitstreams: 2 Tese - Kênia Alves Pereira Lacerda - 2016.pdf: 1930787 bytes, checksum: d2d35012cd70eaa30ae8e03b674c7055 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-01-16T11:01:34Z (GMT). No. of bitstreams: 2 Tese - Kênia Alves Pereira Lacerda - 2016.pdf: 1930787 bytes, checksum: d2d35012cd70eaa30ae8e03b674c7055 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2016-12-06 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / Vitiligo is a skin disease that affects about 1% of the population worldwide. This disease does not cause physical incapacity, but due to its disfiguring appearance, the carriers can present psychological disturbances and impairment in the quality of life. The pathogenesis has not yet been clarified. Recently, evidence has suggested that some gene polymorphisms are associated with risk of developing vitiligo. This study aimed to investigate the Glu298Asp polymorphism of the endothelial nitric oxide synthase gene (eNOS) in a group of patients with vitiligo and to compare the frequencies of the genotypes with a control group from the center-west region of Brazil, as well as assess the quality of life and the perception of stress In individuals with vitiligo and to compare the perception of stress with non-carriers. First study included 51 individuals with vitiligo and 50 controls. The whole blood samples were obtained by collection of peripheral venous blood, then DNA extraction was performed. The genotyping of the Glu298Asp polymorphism was performed by PCR-RFLP analysis. The research on the association of the polymorphism of the eNOS Glu298Asp gene with vitiligo did not present a significant difference in the distribution of genotypes and in the distribution of allelic frequencies between the carriers and the control group. However, in the analysis of the association between the genotypes and the variables location of vitiligo and skin color in the carriers, a higher proportion of TT genotypes was observed in individuals with 100% unisegmentar vitiligo and GT in patients with vitiligo located 68.3% (p = 0.028 ). Also, in the dominant model, a significantly higher proportion of GG genotype was observed in individuals of black color 34.5%, while a higher frequency of moderate-color individuals with GG + TT genotype was observed 54.5% (p = 0.035 ). In the second study, the sample consisted of 102 participants divided into two groups, with vitiligo and without vitiligo. We used the Dermatology Life Quality Index questionnaires and the Perceived Stress Scale. The mean quality of life score for patients with vitiligo was 4.7 ± 5.8, demonstrating a mild impairment in the patients. There was a significant association between quality of life and black skin color (β = 5.64, p <0.001), pattern of involvement of exposed vitiligo (β = 5.22, p <0.001) and perceived stress (β = 0,22 p = 0.033). The perceived stress assessed between the groups presented a mean of 20.7 ± 6.0 and 17.8 ± 7.0 for case and control, respectively. Vitiligo patients presented a high perception of stress when compared to the control group (β = 3.12, p = 0.022). The results pointed out a high perception of stress in vitiligo patients, evidencing that the fact of having vitiligo increases the level of stress. It is known that several factors, such as the genetic profile and ethnicity of the analyzed population, can influence the complex susceptibility to the disease, therefore, more studies are needed to elucidate the role of eNOS gene polymorphism in the pathogenesis of vitiligo. / O vitiligo é uma doença da pele que afeta cerca de 1% da população em todo o mundo. Esta doença não causa incapacidade física, mas devido à sua aparência desfigurante, os portadores podem apresentar distúrbios psicológicos e comprometimento na qualidade de vida. A patogênese ainda não foi esclarecida. Recentemente, evidências sugeriram que alguns polimorfismos de genes estão associados com risco de desenvolvimento do vitiligo. Este estudo objetivou investigar polimorfismo Glu298Asp do gene óxido nítrico sintase endotelial (eNOS) em um grupo de pacientes com vitiligo e comparar as frequências dos genótipos com um grupo controle da região centro oeste do Brasil, também avaliar a qualidade de vida e a percepção de estresse em indivíduos portadores de vitiligo e comparar a percepção de estresse com os não portadores. Primeiro estudo, incluiu 51 indivíduos com vitiligo e 50 pessoas controles. As amostras de sangue total foram obtidas por coleta de sangue venoso periférico, em seguida, foi realizado a extração do DNA. A genotipagem do polimorfismo Glu298Asp foi realizada por análise de PCR-RFLP. A investigação sobre a associação do polimorfismo do gene eNOS Glu298Asp com vitiligo, não apresentou diferença significativa na distribuição dos genótipos e na distribuição das frequências alélicas entre os portadores e grupo controle. Porém, na análise da associação entre os genótipos e as variáveis localização do vitiligo e cor da pele nos portadores, verificou maior proporção de genótipos TT em indivíduos com vitiligo unisegmentar 100% e GT em portadores com vitiligo localizado 68,3% (p = 0,028). Ainda, no modelo dominante, verificou-se uma proporção significativamente maior de genótipo GG em indivíduos da cor negra 34,5%, enquanto se verificou maior frequência de indivíduos da cor morena moderada com genótipo GG+TT 54,5% (p = 0,035). No segundo estudo, a amostra foi composta por 102 participantes divididos em dois grupos, com vitiligo e sem vitiligo. Utilizou-se os questionários Dermatology Life Quality Index e a Perceived Stress Scale. O escore médio de qualidade de vida para os portadores de vitiligo foi de 4,7 ± 5,8, demonstrando um comprometimento leve nos portadores. Verificou-se associação significativa entre qualidade de vida e cor da pele negra (β = 5,64; p < 0,001), padrão de envolvimento do vitiligo exposto (β = 5,22; p < 0,001) e estresse percebido (β = 0,22; p = 0,033). O estresse percebido avaliado entre os grupos apresentou média de 20,7±6,0 e 17,8±7,0 para caso e controle, respectivamente. Portadores do vitiligo apresentaram uma percepção elevada de estresse quando comparado ao grupo controle (β = 3,12; p = 0,022). Os resultados, apontaram uma percepção elevada de estresse nos portadores de vitiligo, evidenciando que o fato de possuir vitiligo aumenta o nível de estresse. Sabe-se que vários fatores, tais como o perfil genético e etnia da população analisada, podem influenciar no complexo susceptibilidade à doença, portanto, mais estudos são necessários para elucidar o papel do polimorfismo do gene da eNOS na patogênese do vitiligo.

Coronary artery disease (CAD), Hypertension (Ht) and Type 2 Diabetes Mellitus (T2DM) are all global health problems. This is particularly evident amongst South Asian population groups. The conventional risk factors do not fully explain the higher prevalence of these diseases among South Asians. The endothelial Nitric Oxide Synthase (eNOS) gene is responsible for the production of Nitric Oxide (NO), which may contribute to the physiology of all three disease states. Endothelial dysfunction (which is characterised by a reduction in basal NO) has been shown to be present in, or prior to all three diseases. Numerous variations exist within the eNOS gene, of these variations three have been shown to have a possible functional effect. The first is the Glu298Asp polymorphism within the exon region of the gene, resulting in an amino acid substitution of Glutamate (Glu) to Aspartate (Asp). The second, known as the T-786C polymorphism, is a thymine to cytosine mutation at position -786 in the promoter region. Finally a VNTR polymorphism in Intron 4 causes either a 4 27bp repeat or a 5 27bp repeat. It is hypothesised that these variations could have an effect on the ability of eNOS to produce NO and thus may increase the risk or contribute to the development of the diseases. Previous studies on these variants have shown conflicting results and further studies are warranted to understand and confirm the role of eNOS gene polymorphisms in cardio-metabolic diseases. There is very limited research into the distributions of these genetic variants and their interaction in diseases processes in North Indian populations. Objectives: 1. To analyse through a case control study three different polymorphisms of the eNOS gene for possible association with Coronary Artery Disease (CAD), Hypertension (Ht) and Type 2 Diabetes Mellitus (T2DM) in North Indian population groups. 2. To statistically evaluate descriptive statistics including; age, gender, smoking, dietary behaviours and lipid parameters for possible influence on disease and potential interaction with genetic polymorphisms. 3. To evaluate linkage disequilibrium between the three eNOS variants and carryout haplotype analysis to work out haplotype risk in different diseases. 4. To analyse through a case control study the deletion variant of the Angiotensin-converting enzyme (ACE) gene for possible association with Coronary Artery Disease (CAD), Hypertension (Ht) and Type 2 Diabetes Mellitus (T2DM) in North Indian population groups. 5. To determine a possible interactive effect of the eNOS polymorphisms with the ACE polymorphism. Subjects and Methods: The Glu298Asp and Intron 4 variants were genotyped using a PCR-RFLP technique, the T-786C variant was genotyped using a real time-PCR technique. The ACE deletion variant was also genotyped using a standard PCR technique. The genotyping was undertaken in a total of 457 CAD patients and 220 matched controls from Lucknow, Uttar Pradesh in North India, 319 T2DM patients and 307 matched controls from Punjab, North India and 210 Ht and 162 matched controls, also from Punjab, North India. Results: CAD: The Glu298Asp was significantly associated with CAD among smokers (TT+GT vs. GG OR=2.84 (CI: 1.61-5.0), p<0.001). The Intron 4 variant was also significantly associated with CAD in a smoking dependent manner (4aa+4ab vs. 4bb OR=0.56 (CI: 0.33-0.96). The T-786C variant showed no overall influence on CAD risk. There was also evidence for both synergistic and haplotypic effects of the eNOS gene on CAD status (haplotype G-C-4b OR=4.76 (CI: 1.43-15.78), p<0.001). The ACE genetic variant was confirmed to be a strong independent risk factor for CAD under a dominant model (OR=2.18 (CI: 1.46-3.25), p<0.001). There was no evidence for an interactive effect between the ACE deletion and any of the three eNOS variants incorporated in the current study. Ht: The Glu298Asp variant was not shown to increase Ht risk, with a reduced risk association found under a recessive model (OR=0.316 (CI:0.089-1.116)), p=0.061). The T-786C variant s role in disease remained unclear with the findings showing a non significant increased risk. The Intron 4 variant was also shown to increase Ht risk, in a non significant manner. Sufficiently powered studies would be required to clarify these possible associations. The combined analysis, using logistic regression and haplotype analysis revealed no significant associations, but there was a possible protective effect of the T-C-4b haplotype (OR=0.46 (CI: 0.21-1.01), p=0.054). The ACE gene variant was confirmed to be a strong independent risk factor for Ht under a recessive model (OR=1.81 (CI: 1.20-2.74), p=0.01). Again there was no evidence for an interactive effect between the ACE deletion and any of the three eNOS variants in hypertension. T2DM: The Glu298Asp variant was found to be associated with T2DM under a dominant model, the protective effect remained significant following adjustment for conventional risk factors and other gene variants (OR=0.407 (CI: 0.231-0.717), p=0.002). The T-786C variant showed no overall influence on T2DM risk. The Intron 4 variant also found no overall influence. Haplotype analysis found the T-T-4b was found to be significantly protective for T2DM (OR=0.41 (CI: 0.26-0.65), p=0.0002). Finally the ACE gene variant was confirmed to be a risk factor for T2DM under a dominant model (OR=2.62 (CI: 1.51-4.54), p=0.001). Overall Conclusions: To conclude, this study successfully identified the frequency of three eNOS gene variants and the ACE deletion variant in three complex diseases within north Indian populations. There is a clear role of the eNOS gene in all three diseases and consequently the genetic variants have susceptible/protective associations. The association with disease was found to be present at an individual level, in association with risk factors and at a haplotypic level. These findings warrant further studies to confirm and untangle the genetics of complex diseases and genetic risk profiles calculations which will contribute to the field of medical genomics/personalised medicare and interventions among North Indian populations.