A hérnia inguinal é uma doença multifatorial que emerge do orifício de Fruchaud, fechado somente pela fáscia transversal. Nos últimos tempos tem sido demonstrado que desordens dos elementos do tecido conjuntivo, como fibras colágenas e elásticas, estão relacionados com a gênese da hérnia inguinal. Estudos prévios demonstraram alterações estruturais e quantitativas das fibras elásticas com o envelhecimento da fáscia transversal, relacionado ao aparecimento de hérnia inguinal a partir da quinta década da vida. Estudos recentes demonstraram associação entre uma mutação pontual do éxon 20 do gene da elastina, componente amorfo das fibras elásticas, e hérnia inguinal em indivíduos do sexo masculino. A fibrilina-1 é o principal componente microfibrilar das fibras elásticas e está relacionada ao surgimento de síndromes genéticas, como Marfan, Ehlers-Danlos e Williams, que também apresentam indivíduos portadores de hérnias. Nesse sentido, o objetivo do presente estudo foi investigar a presença de polimorfismos no gene da fibrilina-1 (FBN1) em indivíduos portadores de hérnia inguinal. Estudou-se o ácido desoxirribonucléico (DNA) genômico de 60 pacientes com hérnia inguinal e 60 controles. Os exons 4, 13, 24, 25, 26, 27, 31 32, 39, 41, 59 e 65 foram amplificados pela Reação em Cadeia da Polimerase e, posteriormente, foram avaliados os polimorfismos em gel de poliacrilamida. Todos os exons estudados apresentaram alguns indivíduos com padrão de bandeamento diverso. O produto de amplificação destes exons foi então avaliado através de seqüenciamento e confrontado com a base de dados do National Human Genome Research Institute. Alterações de inserção e/ou deleção consistentes foram observadas no éxon 27: 1) inserção de uma base entre os codons 1119 e 1120 (GAT -> AGA); 2) inserção de uma base entre os codons 1116 e 1117 (TGT -> CTG): 3) inserção de uma base no códon 1148 (CCC -> CGC); e no éxon 31: inserção de uma base entre os codons 1282 e 1283 (GAG -> CGA). Não houve significância estatística que indique associação entre o gene FBN1 e hérnia inguinal. Entretanto, como o gene da fibrilina-1 é bastante grande (350 kD, dividido em 65 exons) e, em outras doenças como a Síndrome de Marfan mais de 500 mutações já foram descritas, sem haver exons que possuam uma maior responsabilidade pela Síndrome, é possível que outros exons possam estar relacionados com o acometimento da hérnia inguinal. / The inguinal hernia is a multifactorial disease that emerge from the Fruchaud orifice, closed only by the transversalis fascia. Lately it has been showed that disorders on the connective tissue elements, as the collagen and elastic fibers, are related to the inguinal hernia genesis. Previous studies have showed structural and quantitative changes of the elastic fibers at the transversalis fascia with aging, that may be related to inguinal hernia at the fifth decade of life. Recent studies have demonstrated an association among a punctual mutation on exon 20 of the elastin gene, amorfous component of the elastic fibers, and male individuals with inguinal hernia. The fibrillin-1 is the main microfibrillar component of the elastic fibers and is associated to genetic syndromes as Marfan, Ehlers-Danlos and Williams, that also present inidividuals with hernias. Thus, the objective of this study was to investigate the presence of polymorphisms on the fibrillin-1 gene (FBN1) on individuals who developed inguinal hernia. The desoxirribonucleic acid (DNA) of 60 individuals with inguinal hernia and 60 controls have been studied. The exons 4, 13, 24, 25, 26, 27, 31, 32, 39, 41, 59 and 65 were amplyfied by the Polymerase Chain Reaction and later evaluated the polymorphisms on polyacrilamide gel. The amplification product of these exons were evaluated by DNA sequencing and compared to the National Human Genome Research Institute database. Consistent mutations were observed at exon 27: 1) insertion of a base between codons 1119 and 1120 (GAT -> AGA); 2) insertion of a base between codons 1116 and 1117 (TGT -> CTG); 3) insertion of a base on codon 1148 (CCC -> CGC); and at exon 31: insertion of a base between codons 1282 and 1283 (GAG -> CGA). There was no statistic significance that could indicate the association between FBN1 gene and inguinal hernia. However, FBN1 is a large gene (350 kD, shared in 65 exons) and in other disorders as Marfan Syndrome, more than 500 mutations have already been described, without the existence of prevalent exons that have major responsability about the Syndrome, it is possible that other exons could be related to the happening of inguinal hernia.

Based on previous studies, the increasing prevalence of hypertension may be associated with factors such as obesity, dietary salt and fat intake. This study examined the common biochemical and anthropometric markers that are associated with blood pressure elevation, increasing metabolic and haemodynamic derangement in subjects in Hong Kong, and related those phenotypic markers to some genetic polymorphisms relevant to hypertension. / Five hundred and thirty nine Hong Kong Chinese subjects were examined. They were aged from 20 to 60 years, and were hypertensive or normotensive siblings from families with a hypertensive proband, and normotensive controls without a family history of hypertension. The interrelationships between pathophysiological changes and various neurohormones considered relevant to the development of hypertension were investigated. Fasting blood and 24 hour urine samples were collected. Plasma insulin, plasma leptin, plasma renin activity (PRA), serum angiotensin converting enzyme (ACE) activity, aldosterone, 24 hour urine noradrenaline, adrenaline, dopamine and kallikrein were measured. A robust assay for the measurement of urine free cortisol and cortisone and 6beta-hydroxycortisol by an LC-MS/MS method was developed and validated. The ratio between urine free cortisol and cortisone was used as an estimate of the activity of 11beta-hydroxysteroid dehydrogenase type II (11betaHSD2) for cortisol metabolism. These parameters were related to polymorphisms in three genes, the angiotensinogen (AGT) gene M23 5T, the dopamine D1 receptor (DD1R) gene A-48G and the dopamine D2 receptor (DD2R) gene Taq1 A polymorphisms. Analysis of variance was employed for the parameters in the three groups of subjects and for an age-matched sibling pair analysis (using 1 normotensive and 1 hypertensive sibling from each family). Comparisons between parameters were also made after dividing the whole population into 3 groups according to the tertiles of blood pressure. (1) Central (higher waist to hip ratio and waist circumferences) and general (greater body mass index and weight) obesity were found in both hypertensive patients and the normotensive siblings compared to the control subjects. These obesity indices showed strong positive relationships with increased insulin resistance and blood pressure. The obesity indices were also independently associated with systolic and diastolic blood pressure, with central obesity showing the stronger associations. (2) Hypertensives had more adverse lipid profiles, insulin resistance and higher fasting plasma glucose levels. This suggested that the blood pressure elevation in the hypertensives may be mediated through obesity and insulin resistance. (3) Both the hypertensive and normotensive members of sibling pairs had lower noradrenaline and cortisol excretion and higher activity of 11betaHSD2 compared to the normotensive controls. The result showed positive relationships between noradrenaline and increased obesity, insulin resistance and blood pressure, while the relationship between adrenaline and blood pressure was inversed. (4) Lower plasma ACE activity and aldosterone were found in the hypertensives and their siblings than in the normotensive controls. There was a reduction in PRA across the blood pressure tertiles as blood pressure increased. In addition to the higher 11betaHSD2 activity, a negative relationship between aldosterone and blood pressure in hypertensive siblings was observed. These findings may indicate the protective mechanism of these systems in this population. In subjects with the different polymorphisms of AGT M235T, there were no differences in the PRA, serum ACE activity or aldosterone, but lower urine cortisol and kallikrien were found in relation to increasing numbers of the T allele. There was a weak association between the AGT M235T polymorphism and hypertension. (5) Despite the strong correlation of dopamine excretion between hypertensive and normotensive siblings within families, lower dopamine levels were found in the normotensive siblings. A consistent positive relationship was found between urine dopamine and sodium excretion, which supports the concept of the natriuretic effect of dopamine. There was no phenotypic difference found in any of the biochemical parameters in relation to the DDIR A-48G and DD2R Taq1 A polymorphisms, but there were weak associations with blood pressure and these polymorphisms in the sibling study. / The normotensive siblings had metabolic abnormalities similar to but less severe than the hypertensive probands, which suggests that the genetic effects and interacting effect of shared lifestyle and environmental factors with their hypertensive family member may be influential on the healthy siblings. Adaptive changes were seen in some of the blood pressure regulating systems in both the hypertensive probands and the normotensive siblings. The major factors predisposing to the hypertension in these subjects appeared to be obesity and insulin resistance and the adaptive changes were insufficient to compensate for these in the hypertensive subjects. / Chu Ten Wah Tanya. / "March 2006." / Adviser: Brian Tom Linson. / Source: Dissertation Abstracts International, Volume: 68-03, Section: B, page: 1547. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (p. 304-341). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Introdução e Objetivos: A maior prevalência de doença celíaca (DC) em indivíduos com diabetes mellitus tipo I (DM1) já é reconhecida. Ambas as doenças tem causa autoimune, em que os genes HLA classe 2 representam o principal fator genético de risco. Porém, existe uma considerável parcela da população que não manifesta tais doenças e são portadores desses genes. Estudo de associação genômica (GWAS) identificaram polimorfismos de susceptibilidade às duas doenças em genes diferentes do sistema HLA, que poderão auxiliar na compreensão da causa e das suas variabilidades clínicas. Os objetivos desse estudo foram avaliar as frequências dos polimorfismos HLA e não HLA em pessoas como DM1 e com DC e relacionar esses dados com a ocorrência de sintomas gastrointestinais, com a idade do diagnóstico da DM1 e com história alimentar. Métodos: Delineamento transversal, com avaliações retrospectivas e prospectivas, em pessoas com DM1 com e sem DC. Foram realizadas entrevista e revisão de prontuário dos pessoas, seguido de coleta de sangue ou saliva. A pesquisa dos genes RGS1, IL2-IL21, BACH2, TLR7/TLR8 e IL18RAP foi realizada por PCR Real-Time. Os alelos DQA1* 0501 e DQB1* 0201 para DQ2.5 e o alelo DQB1*0302 para DQ8 foram identificados a partir da técnica de genotipagem de HLA Tag-single-nuleotide polymorphism (Tag SNP). Resultados: As frequências alélicas e genotípicas entre 273 pessoas com DM1 sem DC e 39 pessoas com DM1 e DC não apresentaram diferença significativa. A presença de sintoma gastrointestinal foi mais frequente nos portadores dos polimorfismos dos genes RGS1 e IL18RAP. O tempo de aleitamento materno, a idade de introdução do glúten e a idade do diagnóstico da DM1 foram semelhantes entre os grupos. A comparação dos cinco polimorfismos com a combinação dos haplótipos para DQ2.5 e DQ8 não apresentou diferença significativa. Nos 312 indivíduos, com DM1 com e sem DC e nos 66 indivíduos portadores de DC sem DM1 foi identificado alelos DQ2.5 e ou DQ8 em 97% dos casos, enquanto que nos indivíduos com DC sem DM1 identificou-se em 76% dos casos. DQ2.5 foi mais frequente entre pessoascom DC e DQ8 foi mais frequentes entre pessoas com DM1. Conclusões: A presença dos polimorfismos dos genes estudados não modificou a chance do indivíduo com DM1 ter ou não DC. Houve associação dos genes RGS1 e IL18RAP com sintomas gastrointestinais. A pesquisa dos alelos DQ2.5 e DQ8, pela técnica Tag-SNP, permitiu determinar um alto valor preditivo negativo no diagnóstico de DC na população com DM1 e com DC, semelhante ao descrito na literatura com a técnica convencional. / Introduction and Objectives: The higher prevalence of celiac disease (CD) in individuals with diabetes mellitus type I (T1D) is already recognized. Both diseases have autoimmune cause, where HLA genes class 2 represent the major genetic risk factor. However, there is a considerable portion of the population that does not manifest such diseases and are carriers of these genes. Genome-wide association studies (GWAS) have identified susceptibility polymorphisms to both diseases in different genes of the HLA system that may assist in understanding the etiology and in its clinical variabilities. The objectives of this study were to evaluate the frequencies of HLA and non-HLA polymorphisms in patients with T1D and CD, related to the occurrence of gastrointestinal symptoms, the age of diagnosis of T1D and food history. Methods: Mixed design with retrospective and prospective evaluations in patients with T1D with and without DC. They were conducted interview and review of medical records of patients, followed by collecting blood or saliva. The search for genes RGS1, IL21-IL2, BACH2, TLR7 / TLR8 and IL18RAP was performed by Real-Time PCR. The alleles DQA1 * 0501 and DQB1 * 0201 for DQ2.5 and DQB1 * 0302 for DQ8 were identified from the Tag-single-nucleotide polymorphism (tag SNP) genotyping HLA technique Results: The allelic and genotypic frequencies between 273 T1D patients without CD and 39 patients with T1D and CD showed no significant difference. The presence of gastrointestinal symptoms were more frequent in patients with polymorphisms of genes RGS1 and IL18RAP. The duration of breastfeeding, the age of introduction of gluten and the age of diagnosis of T1D were similar between the groups. The comparison of the five polymorphisms with the combination of haplotypes for DQ2.5 and DQ8 showed no significant difference. In 312 individuals with DM1 with and without CD and 66 individuals with CD without T1D was identified alleles DQ2.5 and/or DQ8 in 97% of cases, whereas in individuals with CD without T1D was identified in 76% of cases . DQ2.5 was more frequent among patients with CD and DQ8 was more frequent among patients with T1D Conclusions: The presence of polymorphisms of genes studied did not modify the chance of T1D whether or not DC. There was an association of RGS1 and IL18RAP genes with gastrointestinal symptoms. The survey of DQ2.5 and DQ8 alleles by Tag-SNP technique allowed determining a high negative predictive value in the diagnosis of CD in the population of patients with T1D and DC, similar to that described in the literature with the conventional technique.

Aim. The human organic cation transporter subtype 2 (OCT2) is highly expressed in the renal tubular epithelium and can play an important role in the renal clearance of many drugs and drug-drug interactions occurring in the kidney. The purposes of this study are: (1) to investigate the genetic polymorphisms of OCT2 in Chinese population; (2) to evaluate the potential genotype-phenotype relationship involving OCT2 polymorphism in human subjects; and (3) to identify the SNPs in proximal promoter/enhancer region and assess their functional significance. / Conclusion. The present study demonstrated the existence of genetic polymorphisms of OCT2 gene in the Chinese population and for the first time showed that a ncSNP 808G>T is associated with a reduced renal transport function and can significantly impact the magnitude of drug interactions. Our study also for the first time found that a promoter polymorphism (-1283 T>C) is associated with an altered promoter activity in vitro, but no such relationship was observed with this SNP in the in vivo metformin study. Thus, it was the ncSNP 808G>T but not the -1283T>C in promoter that was associated with variations in the metformin renal clearance. (Abstract shortened by UMI.) / Method. One hundred and twelve Hong Kong Chinese subjects were recruited and their DNA samples were obtained. / Results. A total of 13 SNPs were identified in the coding and surrounding non-coding regions of OCT2, with minor allele frequencies (MAF) ranged from 4.5% to 24.7%. From these SNP data sets, 28 haplotypes were inferred with 4 being the common ones (frequencies ranged from 5.4% to 50.4%). Only one non-synonymous coding region SNP (ncSNP), 808G>T in the exon 4, was observed among all the identified SNPs. Significant differences were observed in the renal clearance of metformin in subjects with different mutation status for this variant. The mean renal tubular clearance (CLt) values of metformin were 8.54 +/- 1.86, 7.72 +/- 0.64, and 6 36 +/- 0.98 ml/kg/min for subjects with GG (n = 6), GT (n = 5) and TT (n = 4) genotypes respectively (P = .043. 1-way ANOVA). After a 6-day cimetidine treatment, a mean decrease of 50.7%, 34.6% and 18.9% in metformin CLt was observed in the GG, GT and TT genotype groups (P =.013, P =.002 and P = .065 respectively compared to metformin alone). The decrease of CLt was significantly lower in the TT genotype group than that in the GG group (P = .027). Five SNPs were identified and 5 haplotypes inferred (frequencies ranged from 2.7% to 38.4%) in promoter/enhancer area. One haplotype, characterized by the presence of -1283 T>C, was associated with a significantly lower luciferase activity in vitro (26.7% decrease in comparison to wild-type, P = .016), but not with metformin CLt in Chinese Subjects. / Wang, Zhijun. / "Aug 2007." / Adviser: Moses Chow. / Source: Dissertation Abstracts International, Volume: 69-02, Section: B, page: 0966. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (p. 148-168). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstracts in English and Chinese. / School code: 1307.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2016-06-21T19:17:07Z No. of bitstreams: 1 tairinezaralopes_dissert.pdf: 1511634 bytes, checksum: 2203122e0b39cf1687115059c3e19447 (MD5) / Made available in DSpace on 2016-06-21T19:17:07Z (GMT). No. of bitstreams: 1 tairinezaralopes_dissert.pdf: 1511634 bytes, checksum: 2203122e0b39cf1687115059c3e19447 (MD5) Previous issue date: 2015-10-29 / Introduction: Thyroid cancer is the most common malignancy of the endocrine system and has been presenting continuous increase in the last years. Studies suggest that folate deficiency in the body decrease DNA repair, resulting in malignant cells changes that alter expression of genes, and may induce several kinds of cancer development. Polymorphisms in genes involved in folate pathway have been investigated as risk factors for susceptibility to cancer, among them MTHFR, MTR, RFC1 and CßS. Objectives: To investigate association of polymorphisms in the MTHFR (C677T), MTR (A2756G), RFC1 (A80G) and CßS (844ins68) genes in risk thyroid cancer in a case-control study; to evaluate the association of polymorphisms with gender, age, alcohol and tobacco consumption, body-mass index in thyroid cancer development; and to evaluated the association between polymorphisms and clinical-histopathological parameters. Methods: This study included 462 individuals (151 patients with thyroid cancer and 311 controls). The peripheral blood was collected and genomic DNA was extracted. The MTHFR (C677T), MTR (A2756G) and RFC1 (A80G) were evaluated by PCR-RFLP and CßS (844ins68) by conventional PCR without enzymatic digestion. For statistical analysis chi-square and multiple logistic regression were used. Results: The results showed that MTHFR C677T (OR=2.87, 95% CI=1.50-5.48, p< 0.01, codominant model), (OR=1.76, 95% CI=1.18-2.64, p< 0.01, dominant model), (OR=2.37, 95% CI=1.28-4.39, p< 0.01, recessive model) and RFC1 A80G (OR: 1.55; 95% CI: 1.02-2.38; p=0.04, recessive model) were associated with thyroid cancer. The alcohol (OR=1.56, 95% CI=1.36-1.89, p< 0.01) and tobacco consumption (OR=1.97, 95% CI=1.28-3.04, p< 0.01) were statistically significant, being associated with increased risk. The MTR A2756G is associated with tumor extension (OR=2.69, 95% CI=1.27-5.71, p< 0.01) and aggressiveness (OR= 4.51, 95% CI=1.67-12.1, p< 0.01). Conclusions: The MTHFR (C677T) and RFC1 (A80G) polymorphisms were involved in risk for thyroid cancer. Additionally, alcohol and tobacco consumption increase risk for disease development. / Introdução: O câncer de tireoide é a neoplasia maligna mais comum do sistema endócrino e vem apresentando contínuo aumento nos últimos anos. Estudos sugerem que a deficiência de folato no organismo diminui a reparação do DNA, resultando em alterações celulares malignas que modulam a expressão gênica, podendo levar ao desenvolvimento de vários tipos de câncer. Polimorfismos em genes envolvidos na via do folato têm sido investigados como fatores de risco para suscetibilidade ao câncer, entre eles, polimorfismos nos genes MTHFR, MTR, RFC1 e CßS. Objetivos: Investigar a associação dos polimorfismos nos genes MTHFR (C677T), MTR (A2756G), RFC1 (A80G) e CßS (844ins68) no risco de câncer de tireoide em um estudo caso-controle; Avaliar a associação dos polimorfismos com o gênero, idade, consumo de álcool e tabaco, índice de massa corpórea (IMC) no desenvolvimento do câncer de tireoide; Avaliar a associação entre os polimorfismos e os parâmetros clínico-histopatológicos do câncer de tireoide. Casuística e Método: Este estudo incluiu 462 indivíduos (151 pacientes com câncer de tireoide e 311 indivíduos controles). Foi coletado sangue periférico e extraído o DNA genômico. Os polimorfismos MTHFR (C677T), MTR (A2756G) e RFC1 (A80G) foram avaliados por meio da PCR-RFLP e o polimorfismo CßS (844ins68) foi analisado por PCR convencional sem corte enzimático. Para análise estatística utilizou-se o teste do qui-quadrado e regressão logística múltipla. Resultados: Os resultados mostraram que os polimorfismos MTHFR C677T (OR=2.87, 95% IC=1.50-5.48, p< 0.01, modelo codominante), (OR=1.76, 95% IC=1.18-2.64, p< 0.01, modelo dominante), (OR=2.37, 95% IC=1.28-4.39, p< 0.01, modelo recessivo) e RFC1 A80G (OR: 1.55; 95% IC: 1.02-2.38; p=0.04, modelo recessivo) estão associados ao câncer de tireoide. O consumo de álcool (OR=1.56, 95% IC=1.36-1.89, p< 0.01) e tabaco (OR=1.97, 95% IC=1.28-3.04, p< 0.01) foram estatisticamente significantes, sendo associados ao aumento do risco. O polimorfismo MTR A2756G está associado à extensão do tumor (OR=2.69, 95% IC=1.27-5.71, p< 0.01) e à agressividade (OR= 4.51, 95% IC=1.67-12.1, p< 0.01). Conclusões: Os polimorfismos MTHFR (C677T) e RFC1 (A80G) estão envolvidos no risco de câncer de tireoide. Adicionalmente, o consumo de álcool e tabaco aumenta o risco de desenvolvimento da doença.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2017-08-03T17:28:33Z No. of bitstreams: 1 adrianaadacruzfurini_tese.pdf: 7663546 bytes, checksum: 1b3a248d18a2d722dab05643a42828b2 (MD5) / Made available in DSpace on 2017-08-03T17:28:33Z (GMT). No. of bitstreams: 1 adrianaadacruzfurini_tese.pdf: 7663546 bytes, checksum: 1b3a248d18a2d722dab05643a42828b2 (MD5) Previous issue date: 2016-08-05 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Introduction: Malaria is one of the mayor cause of morbidity and mortality in tropical and subtropical countries. Objectives: To evaluate the influence of genetic ancestry in the distribution of polymorphisms in genes involved in the immune response and antibody levels against proteins expressed in the merozoite stage of Plasmodium vivax. Material and Methods: To evaluated 90 patients with vivax malaria and 51 non-infected patients from Goianésia do Pará, northern Brazil. Nine single nucleotide polymorphisms (SNPs) in the genes: TNFA, IL-10 INFG were genotyped by PCR-ASO or RFLP-PCR. The genetic ancestry for three ethnic groups (African, European and American Indian) were categorized using 48 INDELs. The responses of specific antibodies against the C-terminal proteins (MSP-119) MSP-1, BPD and AMA-1 of P. vivax were determined by ELISA. Results: There were no differences in ancestry proportions in most SNPs investigated only for TNF-308A allele and European ancestry. No significant association was observed between the allele and genotype frequencies of the SNPs between the groups investigated. There was no significant difference in the levels of IgG antibodies to the studied polymorphisms. Conclusions: These results indicated that the polymorphisms in the TNFA, INFG e IL10 genes can not influence the anti-merozoites immune response of P. vivax. We discussed the immunogenetic profile involved in the humoral immune response in malaria vivax in an endemic area of the Brazilian Amazon. / Introdução: A malária é uma das maiores causas de morbidade e mortalidade em países tropicais e subtropicais. Objetivos: Avaliar a influência da ancestralidade genética na distribuição de polimorfismos em genes envolvidos na resposta imune e os níveis de anticorpos contra proteínas expressas no estágio de merozoíto do Plasmodium vivax. Material e Métodos: Foram avaliados 90 indivíduos com malária vivax e 51 não infectados de Goianésia do Pará, região Norte do Brasil. Nove polimorfismos de nucleotídeo único (SNPs) distribuídos nos genes: TNFA, INFG e IL10 foram genotipados por PCR-ASO ou PCR-RFLP. A ancestralidade genômica para os três grupos étnicos (africana, europeia e ameríndia) foi categorizada com a utilização de 48 INDELs. As respostas de anticorpos específicos contra as proteínas C-terminal (MSP-119) da MSP-1, da DBP e da AMA-1 do P. vivax foram determinadas por ELISA. Resultados: Não houveram diferenças nas proporções de ancestralidade na maioria dos SNPs investigados, apenas para o alelo TNF-308A e a ancestralidade europeia. Nenhuma associação significativa foi observada entre as frequências alélicas e genotípicas dos SNPs entre os grupos investigados. Não foi encontrada diferença significativa nos níveis de anticorpos IgG em relação aos polimorfismos estudados. Conclusões: Esses resultados ressaltam que os polimorfismos nos genes TNFA, INFG e IL10 não influenciam na resposta imune anti-merozoítos do P. vivax. Discutimos o perfil imunogenético envolvido na resposta imune humoral na malária vivax em região endêmica da Amazônia brasileira

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2017-09-29T18:21:41Z No. of bitstreams: 1 camilaravazzigauch_dissert.pdf: 1221932 bytes, checksum: 8184414238eac76d3bb5fced2b86cc83 (MD5) / Made available in DSpace on 2017-09-29T18:21:41Z (GMT). No. of bitstreams: 1 camilaravazzigauch_dissert.pdf: 1221932 bytes, checksum: 8184414238eac76d3bb5fced2b86cc83 (MD5) Previous issue date: 2015-12-03 / Introduction: The HLA (Human Leukocyte Antigen) molecules are proteins encoded by genes highly polymorphic and are involved in the immune response process. Polymorphisms of HLA genes differ between populations, both in frequency and in the presence or absence of specific alleles and haplotypes. Considering that the distribution of organs for transplant depends on HLA matching between donor and recipient, the knowledge and determination of the HLA polymorphism are of great importance in the process of allocation of organs for transplantation. Moreover, the knowledge of the HLA diversity is an important tool for studies of the origin of populations. Objectives: This study aimed to characterize the allele and haplotype frequencies of HLA-A, -B, and -DRB1 in a cohort of renal transplant candidates populations in the region of São José do Rio Preto (State of São Paulo), to compare the allele frequencies between Caucasian and Black in that region, as well as to compare these frequencies with different Brazilian populations reported. Materials and Methods: The HLA-A, -B, and -DRB1 allele and haplotypes frequencies were analyzed in a sample of 2.624 individuals and classified according to the ethnic group (2.347 Caucasians and 277 Blacks). The HLA class I (A, B) and class II (DRB1) specificities were determined by Complement-Dependent Microlymphocytotoxic (CDC) and Polymerase Chain Reaction/Sequence Specific Priming (PCR-SSP) methods, respectively. Results: All loci studied were in Hardy–Weinberg Equilibrium (p>0.05). Twenty-one HLA-A, 34 HLA-B and 13 HLA-DRB1 allelic groups were identified. The most frequent alleles for each locus were HLA-A*02, HLA-B*35, and HLA-DRB1*11. The most frequent haplotypes found were A*01 B*08 DRB1*03 among Caucasians and A*29 B*44 DRB1*07 among Blacks. Conclusions: The most common alleles for each locus among the renal transplant candidates were A*02, B*35 and DRB1*11. The most common haplotype was A*01 B*08 DRB1*03. The same haplotype was the most frequent in Caucasoid sample while the haplotype A*29 B*44 DRB1*07 was the most common in the Blacks sample. / Introdução: As moléculas HLA (Human Leucocyte Antigens) são proteínas codificadas por genes altamente polimórficos e estão envolvidas no processo de resposta imunológica. Os polimorfismos dos genes HLA diferem entre as populações, tanto na frequência como na presença ou ausência de alelos e haplotipos específicos, Considerando-se que a distribuição de órgãos para transplante depende da compatibilidade HLA entre doador e receptor, o conhecimento e determinação do polimorfismo HLA são de grande importância no processo de alocação de órgãos para transplantes, além de ser uma importante ferramenta em estudos populacionais. Objetivos: 1) Determinar as frequências alélicas para os locus HLA-A, -B e -DRB1 em uma amostra de candidatos a transplante renal no Brasil. 2)Determinar os haplótipos HLA mais freqüentes nessa amostra. 3) Comparar as diferenças de frequências alélicas e haplotípicas entre os grupos de caucasóides e negros da população analisada. Materiais e Métodos: As frequências alélicas e haplotípicas para os locus HLA-A, -B e -DRB1 foram analisadas em uma amostra de 2.624 candidatos a transplante renal e classificadas de acordo com o grupo étnico (2.347 Caucasóides e 277 Negros). As especificidades HLA de classe I (AB) e de classe II (DR) foram determinadas de acordo com a técnica Microlinfocitotóxica Dependente de Complemento (CDC) e Polymerase Chain Reaction - Sequence-specific Primers (PCR-SSP), respectivamente. Resultados: Considerando a amostra total, todos os loci estudados estavam em equilíbrio de Hardy-Weinberg (p>0,05). Foram identificados 21 grupos de alelos para o locus HLA-A, 34 para HLA-B e 13 para HLA-DRB1. Os alelos mais freqüentes para cada locus foram HLA-A*02, HLA-B*35 e HLA-DRB1*11. O haplótipo mais freqüente foi A*01 B*08 DRB1*03 entre a amostra de Caucasóides e A*29 B*44 DRB1*07 entre a amostra de Negros. Conclusões: Os alelos HLA mais freqüentes na população de candidatos a transplante renal foram HLA-A*02, HLA-B*35 e HLA-DRB*11. O haplótipo mais comum foi A*01 B*08 DRB1*03. Esse mesmo haplótipo foi o mais frequente na amostra de Caucasóide da população analisada enquanto que, A*29 B*44 DRB1*07 foi o mais comum na amostra de Negros.

Submitted by Fabíola Silva (fabiola.silva@famerp.br) on 2018-02-06T18:02:49Z No. of bitstreams: 1 analiceandreolidasilva_dissert.pdf: 2107238 bytes, checksum: 4f05811d9d83884b9b7412042055b250 (MD5) / Made available in DSpace on 2018-02-06T18:02:50Z (GMT). No. of bitstreams: 1 analiceandreolidasilva_dissert.pdf: 2107238 bytes, checksum: 4f05811d9d83884b9b7412042055b250 (MD5) Previous issue date: 2017-02-03 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Introduction: Down syndrome (DS) is a genetic condition characterized by the presence of three copies of chromosome 21. In most cases, the extra chromosome 21 is of maternal origin due to errors in chromosomal segregation during meiosis. Advanced maternal age at conception is the main risk factor for DS. However, studies suggest that alterations in genes involved in folate metabolism, such as MTHFR (Methylenetetrahydrofolate redutase), may increase the risk for DS regardless maternal age, due to DNA hypomethylation and, consequently, abnormal chromosomal segregation. MTHFR expression is regulated by microRNAs (miRNAs), and polymorphisms in miRNAs may also modify this metabolism with consequences on chromosome disjunction. Objective: To assess whether the presence of MTHFR rs4846048 and rs4846049 and precursor hsa-mir-149rs2292832 polymorphisms is associated with maternal risk for the occurrence of offspring with DS. Casuistic and Methods: In the present study, 167 mothers of individuals with free trisomy 21, and 296 women without previous history of abortion, mothers of individuals without syndrome and without malformation were examined. Polymorphisms were evaluated by real-time polymerase chain reaction (PCR) using the TaqMan® SNP Genotyping Assays (AppliedBiosystems®) commercial assays. Multiple logistic regression analyzes were performed for the polymorphisms in the dominant and recessive models using the Minitab v. 16.0 program. Genotypic combination analysis was performed using the Fisher exact test in the dominant model. Analysis of the haplotypes of the MTHFR polymorphisms rs4846048 and rs4846049 was performed using the Haploview v. 5.0 program. Maternal age equal or greater than to 35 years old as a risk factor for DS was analyzed using binary logistic regression. Results: Maternal age equal to 35 years old or greater was considered a risk factor for offspring with DS (P <0.0001; OR = 9.38; 95% CI = 5.70-15.45). The maternal polymorphisms rs4846048 and rs4846049 of the MTHFR gene were not associated with the occurrence of offspring with DS, regardless of maternal age. Analyzes of combined genotypes of the MTHFR rs4846048, MTHFR rs4846049 and hsa-mir-149 rs2292832 polymorphisms, as well as the MTHFR gene haplotypes, also showed no association with maternal risk for DS. An increased risk for the occurrence of offspring with DS was observed for women under 35 years old at the time of conception and carriers of the TT genotype of hsa-mir-149 rs2292832 polymorphism (OR = 2.02, 95% CI = 1.06 - 3.83). Conclusion: There is no evidence of association between maternal polymorphisms MTHFR rs4846049 and rs4846048 and risk for the occurrence of SD offspring. However, an increased maternal risk for DS is observed in women with maternal age under 35 years old and carriers of the TT genotype of the hsa-mir-149 rs2292832 polymorphism. / Introdução: A síndrome de Down (SD) é uma condição genética caracterizada pela presença de três cópias do cromossomo 21. Na maioria dos casos, o cromossomo 21 extra é de origem materna e é originado devido a erros na segregação cromossômica durante a meiose. A idade materna avançada no momento da concepção representa o principal fator de risco para SD. Entretanto, estudos sugerem que alterações em genes envolvidos no metabolismo do folato, como o MTHFR (Metilenotetrahidrofolato redutase), podem aumentar o risco materno para prole com SD, independente da idade, por resultarem em hipometilação do DNA e, consequentemente, em segregação cromossômica anormal. A expressão de MTHFR é mediada por microRNAs (miRNAs), assim polimorfismos em miRNAs também podem alterar esse metabolismo com consequências na disjunção cromossômica. Objetivo: Avaliar se a presença dos polimorfismos do gene MTHFR rs4846048 e rs4846049 e do pré-miRNA hsa-mir-149 rs2292832 está associada com o risco materno para a ocorrência de prole com SD. Casuística e métodos: Foram avaliadas 167 mães de indivíduos com trissomia livre do cromossomo 21 e 296 mulheres sem história de aborto prévio, mães de indivíduos sem a síndrome e sem malformação. Os polimorfismos foram avaliados pela técnica de discriminação alélica por reação em cadeia da polimerase (PCR) em tempo real utilizando-se os ensaios comerciais TaqMan® SNP Genotyping Assays (AppliedBiosystems®). As análises de regressão logística múltipla foram realizadas para os polimorfismos nos modelos dominante e recessivo utilizando o programa Minitab v. 16.0. A análise de combinação genotípica foi realizada utilizando o teste exato de Fisher, no modelo dominante. A análise dos haplótipos dos polimorfismos MTHFR rs4846048 e rs4846049 foi realizada utilizando o programa Haploview v. 5.0. A idade materna maior ou igual a 35 anos como fator de risco para a SD foi analisada por meio de regressão logística binária. Resultados: A idade materna igual ou maior que 35 anos foi considerada um fator de risco para prole com SD (P<0,0001; OR=9,38; IC 95%=5,70-15,45). Os polimorfismos rs4846048 e rs4846049 do gene MTHFR não foram associados com a ocorrência de prole com SD, independente da idade materna. As análises de genótipos combinados dos polimorfismos MTHFR rs4846048, MTHFR rs4846049 e hsa-mir-149 rs2292832, bem como dos haplótipos do gene MTHFR, também não evidenciaram associação com risco materno para SD. Um risco aumentado para a ocorrência de prole com SD foi observado para mulheres com idade abaixo de 35 anos no momento da concepção, portadoras do genótipo TT do polimorfismo hsa-mir-149 rs2292832 (OR = 2,02; IC 95% = 1,06 - 3,83). Conclusão: Na casuística avaliada não há evidências de associação entre os polimorfismos maternos MTHFR rs4846049 e rs4846048 e risco para a ocorrência de prole SD. Entretanto, um risco materno aumentado para SD é observado em mulheres com idade materna abaixo de 35 anos portadoras do genótipo materno TT do polimorfismo hsa-mir-149 rs2292832.

Submitted by Suzana Dias (suzana.dias@famerp.br) on 2018-11-09T18:03:41Z No. of bitstreams: 1 ChristianeAyo_tese.pdf: 8833199 bytes, checksum: e3a470634a8f41b17966d4b370464f0c (MD5) / Made available in DSpace on 2018-11-09T18:03:41Z (GMT). No. of bitstreams: 1 ChristianeAyo_tese.pdf: 8833199 bytes, checksum: e3a470634a8f41b17966d4b370464f0c (MD5) Previous issue date: 2017-06-30 / Fundação de Amparo à Pesquisa do Estado de São Paulo - FAPESP / Ocular toxoplasmosis, characterized by an intraocular inflammation, is the most common clinical manifestation of toxoplasmosis, the infectious disease caused by the protozoan Toxoplasma gondii. The lesions can affect the macula and other layers of the retina and the choroid, resulting in retinochoroiditis with different degrees of ocular involvement. Chagas disease is resulting from infection by the protozoan Trypanosoma cruzi. After 20 years of infection, about 30% develop chronic Chagas heart disease, which is clinically manifested by malignant ventricular arrhythmia, thromboembolism, sudden cardiac death, and chronic heart failure. Ten per cent of Chagas patients present the digestive form of the disease, characterized mainly by dilatations of the esophagus and/or colon, due to the denervation process. The progression of the infection, as well as the development of the different clinical forms and different degrees of severity may be related to genetic characteristics of the pathogen and the host. Among the factors related to the host, the immunological response is of special interest with genetic markers playing an important modulating role in this context as they may contribute to the pathogenesis or resistance in the clinical course of these infections. Objective: The present study aimed to verify the hypothesis that KIR genes, their HLA ligands and MICA gene polymorphisms are associated with ocular toxoplasmosis (OT) and the different clinical forms of Chagas disease. Patients and Methods: This study included 297 patients with toxoplasmosis, 148 clinically diagnosed with OT and 149 without OT. Moreover, 267 patients with Chagas disease were enrolled: 78 clinically diagnosed with the digestive form of the disease, 107 with the cardiac form and 82 with the mixed form. The ELISA technique was used to confirm infection by T. gondii and T. cruzi. Polymorphisms of the KIR and MICA genes were identified by PCR-SSOP and nested PCR. Continuous variables were compared using the unpaired t test and the Chi-square test with Yates correction or the Fisher's exact test were used compare the other results. Results: In relation to the toxoplasmosis, MICA genotypes and alleles did not differ between patients with and without OT, or between patients with the primary or recurrent manifestations of the disease. KIR3DS1 gene was positively associated with the development of OT. KIR activating genes along with their HLA ligands (KIR3DS1+/Bw4-80Ile+, KIR2DS1+/C2+ and KIR3DS1+/Bw4-80Ile+) were associated with susceptibility to OT and both its primary and recurrent clinical manifestations. The inhibitory pairs - KIR2DL3/2DL3-C1/C1 and KIR2DL3/2DL3-C1 - were associated with resistance to OT and its clinical manifestations, whereas the combination KIR3DS1-/KIR3DL1+/Bw4-80Ile+ was a protection factor for the development of OT and, in particular, against recurrent manifestations. As for Chagas disease, The MICA-129met allele was associated with the development of left ventricular systolic dysfunction (LVSD) in patients with chronic chagasic cardiomyopathy, while the MICA-129val allele was associated with a protection of developing LVSD. In particular, the MICA-129 met/met homozygous haplotype was associated with the development of severe LVSD and the MICA-129 val/val homozygous genotype protected against this condition. It was also possible to demonstrate that the haplotype MICA*008~HLA-C*06 and the combination between KIR genes and their HLA ligands - KIR2DS2-/KIR2DL2-/KIR2DL3+/C1+ - were associated with susceptibility for the digestive clinical form of Chagas disease. Conclusions: Our results demonstrate that KIR genes may influence both OT and the clinical digestive form of Chagas disease, whereas the MICA gene may influence the clinical forms of Chagas disease, but not the development of OT. / A toxoplasmose ocular, caracterizada por uma inflamação intraocular, é a manifestação clínica mais comum da toxoplasmose, doença infecciosa causada pelo protozoário Toxoplasma gondii. As lesões podem afetar a mácula, as diversas camadas da retina e a coróide resultando em retinocoroidite com diferentes graus de comprometimento ocular. A doença de Chagas é resultante da infecção pelo protozoário Trypanosoma cruzi. Após 20 anos de infecção, cerca de 30% dos pacientes desenvolvem a cardiopatia chagásica crônica, que se manifesta clinicamente por arritimia ventricular malígna, tromboembolismo, morte súbita cardíaca e insuficiência cardíaca crônica. Dez por cento dos pacientes apresentam a forma digestiva, caracterizada principalmente por dilatações do esôfago e/ou cólon devido ao processo de denervação. A progressão de uma infecção, assim como o desenvolvimento de diferentes formas clínicas e diferentes graus de gravidade, podem estar relacionadas com as características genéticas do patógeno e do hospedeiro. Dentre os fatores relacionados ao hospedeiro, a resposta imunológica desperta um interesse especial e os marcadores genéticos exercem importante papel modulador neste contexto, pois podem contribuir para patogênese ou resistência do curso clínico dessas infecções. Objetivo: O presente estudo teve como objetivo verificar a hipótese de que os genes KIR, seus ligantes HLA e o polimorfismo do gene MICA estão associados à toxoplasmose ocular (TO) e às diferentes formas clínicas da doença de Chagas. Casuística e métodos: Foram incluídos no estudo 297 pacientes com toxoplasmose, 148 clinicamente classificados com TO e 149 sem TO. Também participaram deste estudo 267 pacientes com doença de Chagas, 78 clinicamente classificados com a forma digestiva da doença, 107 com a forma cardíaca, 82 com a forma mista. O teste de ELISA foi realizado para confirmar as infecções por T. gondii e T. cruzi. Os polimorfismos destes genes foram identificados por PCR-SSOP e PCR nested. As variáveis contínuas foram comparadas utilizando o teste t não pareado. Para comparação dos demais resultados foram realizados o Teste Qui-quadrado com correção de Yates ou o Teste Exato de Fisher. Resultados: Em relação à toxoplasmose, alelos e genótipos MICA não diferiram entre os pacientes com e sem TO, nem entre os pacientes com a manifestação primária ou recorrente da doença. O gene KIR3DS1 foi associado positivamente com o desenvolvimento da TO. Genes KIR ativadores juntamente com os seus ligantes HLA (KIR3DS1+/Bw4-80Ile+ e KIR2DS1+/C2+ + KIR3DS1+/Bw4-80Ile+) foram associados com suscetibilidade à TO e às suas manifestações clínicas primária e recorrente. Os pares inibidores - KIR2DL3/2DL3-C1/C1 e KIR2DL3/2DL3-C1 – foram associados com a resistência à TO e suas manifestações clínicas, enquanto que a combinação KIR3DS1-/KIR3DL1+/Bw4-80Ile+ foi associada como fator de proteção para o desenvolvimento da TO e, em particular, para a manifestação recorrente. Quanto à doença de Chagas, o alelo MICA-129met foi associado como fator de risco para o desenvolvimento da disfunção sistólica ventricular esquerda (DSVE) em pacientes com cardiopatia chagásica crônica, enquanto que o alelo MICA-129val foi associado com a proteção ao desenvolvimento da DSVE. Em especial o haplótipo homozigoto MICA-129 met/met foi associado com o desenvolvimento da DSVE grave e o genótipo homozigoto MICA-129 val/val foi associado com a proteção desta condição. Também foi possível demonstrar que o haplótipo MICA*008~HLA-C*06 e a combinação entre KIR e seus ligantes HLA - KIR2DS2-/KIR2DL2-/KIR2DL3+/C1+ - foram associados como fatores de suscetibilidade à forma clínica digestiva da doença de Chagas. Conclusões: Nossos resultados demonstram que os genes KIR podem exercer influência tanto na toxoplasmose ocular quanto na forma clínica digestiva da doença de Chagas, enquanto que MICA pode exercer influência nas formas clínicas da doença de Chagas, mas não no desenvolvimento da toxoplasmose ocular.

Diss. (sammanfattning) Göteborg : Göteborgs universitet, 2009. / Härtill 5 uppsatser.