A enzima Paraoxonase-1 (PON1) possui atividades paraoxonase, arilestearase e lactonase, entre outras. É a mais estuda da família das PONs que é composta pela PON1, PON2 e PON3. Sugere-se, que todas atuam inibindo o processo de peroxidação lipídica de moléculas como a lipoproteína de baixa densidade (LDL) e alta densidade (HDL), caracterizando assim um possível papel anti-aterogênico. O gene da PON1 apresenta dois sítios polimórficos, com a troca de uma Gln192Arg (Q/R) e Met55Leu, que estão associados com diferenças na atividade e concentrações séricas da enzima. Por sua vez, indivíduos soropositivos para o HIV-1 apresentam alterações do metabolismo lipídico, que poderiam estar associados a alterações na atividade da PON1 e a terapia antirretroviral (TARV) com inibidores de protease (IP). O objetivo do estudo foi determinar as atividades séricas da PON1 e da arilestearase (ARE), e as freqüências alélicas dos polimorfismos genéticos da PON1 192QR e 55LM, e ainda, avaliar a correlação destes parâmetros com as alterações lipídicas em indivíduos soropositivos para o HIV-1 tratados com IP. No período de Setembro de 2009 até Junho de 2012, 174 indivíduos soropositivos e 46 soronegativos para o HIV-1 foram estudados. Foi realizada a genotipagem dos polimorfismos da PON1 192QR e 55LM através de PCR-RFLP. A atividade sérica da PON1/ARE foi avaliada por espectrofotometria empregando-se como substratos o paraoxon e o fenilacetato, respectivamente. O RNA-HIV-1 foi quantificado pelo método NASBA, e os linfócitos T-CD4+ e T-CD8+ por citometria de fluxo. Os níveis séricos de colesterol total, HDL, LDL, triglicérides (TG), ApoA1 e ApoB100 foram determinados e os anticorpos IgG anti-oxLDL por ELISA. A atividade sérica da PON1 foi inferior nos grupos de soropositivos, p<0,05, porém, a atividade ARE não apresentou diferenças entre os grupos estudados, p>0,05. Ambas as atividades não apresentaram relação com os genótipos PON1 192QR e 55LM, e estes genótipos apresentaram uma freqüência alélica semelhante ao grupo de soronegativos. Os níveis séricos de TG foram superiores nos grupos de soropositivos com TARV, p<0,05, enquanto o grupo tratado com IP apresentou níveis séricos de HDL e Apo-A1 inferiores aos demais grupos, p<0,05. Níveis séricos de Apo-B100, IgG anti-oxLDL, e o índice de risco aterogênico foram superiores no grupo tratado com IP, p<0,05. Concluí-se, que indivíduos soropositivos para o HIV-1 apresentaram alterações no metabolismo lipídico, principalmente nos tratados com IP, que adicionalmente apresentaram um maior índice de risco aterogênico e maiores níveis de anticorpos IgG anti-oxLDL. Estas alterações não apresentaram relação com os polimorfismos PON1 192QR e 55LM da PON1, e demonstraram que a atividade da enzima PON-1 esta diminuída em indivíduos soropositivos para o HIV-1 / The enzyme Paraoxonase-1 (PON1) has paraoxonase (PON), arylesterase (ARE) and lactonase activities, among others. It is the most studied member of PON family which is composed of PON1, PON2 and PON3. It is suggested that all members acts by inhibiting the peroxidation of lipid molecules as the low-density lipoprotein (LDL) and high-density lipoprotein (HDL), characterizing a potential anti-atherogenic effect. The PON1 gene has two mainly polymorphic sites, with an exchange of Gln192Arg (Q/R) and Met55Leu (L/M), which are associated with differences in activity and serum concentrations of the enzyme. In turn, seropositive individuals for HIV-1 show changes in lipid metabolism, which could be associated with changes in the activity of PON1 and highly active antiretroviral therapy (HAART) with protease inhibitors (PI). The aim of this study was to determinate the serum PON and ARE activities of PON1, the allele frequencies of PON1 192QR and PON1 55LM genetic polymorphisms and evaluate the correlation between these parameters and lipid abnormalities in seropositive patients for HIV-1 treated with IP. In the period from September 2009 until June 2012, 174 seropositive individuals and 46 soronegative individuals for HIV-1 were studied. We performed PON1 192QR and 55LM genotyping by PCR-RFLP. Serum activities PON and ARE of PON1 were evaluated by spectrophotometry using paraoxon and phenylacetate, respectively, as substrates. The HIV-1-RNA was quantified by the NASBA method, and lymphocytes T-CD4+ and T-CD8+, by flow cytometry. Serum levels of total cholesterol, HDL, LDL, triglycerides (TG), apoA1 and ApoB100 were determined. IgG anti-oxLDL antibodies were quantified by ELISA. The serum PON1 activity was lower in the seropositive group, p<0.05, however, ARE activity did not differ between groups, p>0.05. Both activities had no relation with the PON1 192QR and PON1 55LM genotype, and these individuals showed an allele frequency similar to the seronegative group. Serum levels of TG were higher in groups of HIV-positive with HAART, p<0.05, while the IP-treated group showed serum levels of HDL and ApoA1 lower than other groups, p <0.05. Serum levels of ApoB100, IgG anti-oxLDL antibodies, and atherogenic risk indices were higher in the group treated with PI, p<0.05. It was concluded that individuals HIV-1-infected showed changes in lipid metabolism, especially in those treated with IP, which additionally showed a higher rate of atherogenic risk and higher levels of IgG anti-oxLDL antibodies. These changes did not correlated with PON1 192QR and 55LM polymorphisms and demonstrated that the activity of PON1enzyme is decreased in individuals seropositive for HIV-1

Stroke is the third leading cause of death. Atherosclerosis in the carotid arteries is a risk factor for ischemic stroke. Oxidized low density lipoprotein (LDL) plays a central role in the progression of atherosclerosis. Human paraoxonase 1 (PON1), a high-density lipoprotein (HDL) associated serum esterase/lactonase, protects HDL and LDL from oxidative modifications. Thus, PON1 is protective against the development of atherosclerosis. PON1 gene has two functional coding region (192Q/R and 55L/M) and one promoter region (&amp / #8722 / 107T/C) polymorphism that affect the catalytic efficiency and levels of the enzyme, respectively. In this study, the aim was to determine the importance of PON1 genetic polymorphisms and activity as risk factors for ischemic stroke. The study population was comprised of 172 unrelated adult Caucasian patients with acute hemispheric ischemic stroke and 105 symptom-free controls. Serum and total blood samples were obtained from G&uuml / lhane Military Medical Academy Hospital Neurology Department, Ankara. Hypertension and diabetes were 2 times more common and HDL-C was significantly lower among patients compared to controls. Logistic regression analysis revealed hypertension and smoking to be significant predictors of stroke. Serum PON1 activities towards three substrates, paraoxon (paraoxonase activity / PON), phenyl acetate (arylesterase activity / ARE) and diazoxon (diazoxonase activity), which were measured by spectrophotometric methods, were found to be lower in stroke patients compared to controls. PON and PON/ARE were negatively associated with ischemic stroke by use of logistic regression analysis. PON/ARE was 1.26 times protective against stroke. The frequencies of the risky alleles 192R, 55L and &amp / #8722 / 107T were increased in the patient group. Frequency of the 55L allele of PON1 was significantly increased among patients (0.690) compared to controls (0.628 / P=0.003). Logistic regression analysis revealed PON1 55LL genotype to be associated with a 1.8-fold increase in the risk of ischemic stroke versus control status. Prevalence of triple combined haplotype QRLMTC was significantly lower in stroke patients (4.1%) when compared to controls (11.4% / P=0.019). The combined heterozygote haplotype had around 7 times increased protective effect against stroke in the overall population and 10 times protective effect in the elderly population. The low expressor genotype &amp / #8722 / 107TT was associated with almost 2 times increased risk for stroke in elderly. 192R allele of PON1 represented 1.554 times increased risk for ischemic stroke in hypertensives relative to normotensives. Furthermore, the risk of hypertensive individuals having ischemic stroke was highest in the 192RR group (Odds Ratio / OR=7), followed by 192QR heterozygotes (OR=2.18), and the risk decreased to insignificant levels in 192QQ individuals. 192R allele constituted a 1.55 times increased risk in diabetics. 55L allele was associated with a 1.66 times increased risk of stroke in hypertensives and a 2.6 times increased risk for stroke in diabetics relative to non-diabetics. PON1 &amp / #8722 / 107T allele also represented a 1.35 times risk for stroke in hypertensives.

La present tesi avalua els efectes de certs factors ambientals sobre la paraoxonasa1 (PON1), enzim antioxidant, possiblement protector enfront les malalties cardiovasculars, a través de dos estudis d'intervenció i un de transversal. En primer lloc, el tractament amb simvastatina dels pacients amb hipercolesterolèmia familiar, que presentaven una activitat paraoxonasa baixa, s'associava a un increment de l'activitat fins a valors similars als d'individus normolipèmics, independentment dels polimorfismes PON1-55 o PON1-192. En segon lloc, l'entrenament físic s'associava a un augment de l'activitat paraoxonasa en els individus QQ i una disminució de la mateixa en els portadors de l'al·lel R pel polimorfisme PON1-192. L'increment de l'activitat paraoxonasa immediatament després de l'exercici físic agut era seguit per una disminució subseqüent de l'activitat. La recuperació dels nivells basals d'activitat paraoxonasa a les 24h de l'exercici físic agut es donava en els individus QQ independentment del seu estat d'entrenament, i en els individus portadors de l'al·lel R només quan estan entrenats. En tercer lloc, el consum elevat d'àcid oleic comportava un augment de la concentració de c-HDL i de l'activitat paraoxonasa en els homes portadors dels genotips QR i RR del polimorfisme PON1-192, respectivament.Paraules claus: paraoxonasa, PON1, genotips, simvastatina, hipercolesterolèmia familiar, interacció gen-dieta, lipoproteïna d'alta densitat (HDL), exercici físic agut, entrenament físic, estrès oxidatiu, àcid oleic, oli d'oliva, peròxids lipídics, malaltia cardiovascular. / The present thesis evaluates some environmental factor effects on paraoxonase1 (PON1), an possibly protective against cardiovascular disease antioxidant enzyme, through two intervention studies and a cross-sectional one. First, treatment with simvastatin of the familial hypercholesterolemic patients, which had low paraoxonase activity, was associated with an increase in the activity to values similar to the normolipemic ones, regardless of the PON1-55 or PON1-192 polymorphisms. Second, Regular exercise was associated with an increase in PON1 activity in QQ subjects and with a decrease in R carriers. Increased PON1 activity immediately after a bout of exercise was subsequently followed by a decrease of activity. The recovery of the basal PON1 activity levels at 24 h was found in QQ subjects regardless of their training status and in trained R carriers, but not in untrained R carriers. Third, high oleic acid intake was associated with increased HDL cholesterol and PON1 activity levels only in men who were QR and RR of the PON1-192 polymorphism, respectively.

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-16T14:43:20Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabriela_de_Lemos_Uliano.pdf: 1271320 bytes, checksum: e96f33052e7efd3711ebdbc9ea9e1880 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-16T20:24:24Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabriela_de_Lemos_Uliano.pdf: 1271320 bytes, checksum: e96f33052e7efd3711ebdbc9ea9e1880 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-16T20:24:38Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabriela_de_Lemos_Uliano.pdf: 1271320 bytes, checksum: e96f33052e7efd3711ebdbc9ea9e1880 (MD5) / Made available in DSpace on 2018-05-16T20:24:38Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabriela_de_Lemos_Uliano.pdf: 1271320 bytes, checksum: e96f33052e7efd3711ebdbc9ea9e1880 (MD5) Previous issue date: 2018-02-16 / Sem bolsa / A infância é um período vulnerável para o início das lesões endoteliais, as quais podem ser agravadas pela exposição a fatores nutricionais de risco. Por outro lado, está comprovado que uma elevada concentração de lipoproteína de alta densidade (HDL) protege contra o surgimento destas lesões, refletindo em menor risco de desenvolvimento da doença cardiovascular (DCV). A principal enzima antioxidante e antiinflamatória que contribui com a propriedade antiaterogênica atribuída ao HDL é a paraoxonase 1 (PON1). Sua atividade sérica pode ser influenciada por diversos parâmetros incluindo polimorfismos genéticos, fatores nutricionais, estilo de vida, idade, fármacos, estado de saúde e estresse oxidativo. O objetivo deste estudo foi investigar a atividade da PON1 e sua associação a polimorfismos genéticos, fatores bioquímicos e nutricionais em crianças de 5 a 7 anos de idade atendidas em ambulatórios de pediatria de duas cidades do sul do Rio Grande do Sul. Foi realizado um estudo transversal no período de Janeiro de 2014 a Fevereiro de 2015. O consumo alimentar foi avaliado pelo questionário do SISVAN. Medidas de peso, altura e circunferência da cintura da criança foram utilizadas para avaliar o estado nutricional. Uma amostra de sangue foi coletada e utilizada para avaliação do perfil lipídico e da atividade arilesterase da PON1, medida utilizando fenilacetato como substrato. A amostra biológica também serviu para a determinação do polimorfismo PON1 C(-107)T, realizada pela amplificação por reação em cadeia da polimerase (PCR) seguida de digestão com enzima de restrição. Os dados coletados foram analisados no programa Stata versão 12, assumindo-se um nível de significância de 5%. A mediana da atividade arilesterase da PON1 foi de 89,4kU/L, semelhante entre meninos e meninas. A análise do polimorfismo revelou que o genótipo mais frequente foi o heterozigoto CT (52,1%). O polimorfismo se mostrou em equilíbrio de Hardy-Weinberg. O genótipo CC mostrou valores mais elevados de atividade enzimática do que os apresentados pelo genótipo TT (p<0,01). Níveis desejáveis de HDL relacionaram-se a um incremento significativo da enzima na população total. As concentrações desta lipoproteína foram significativamente maiores apenas nas crianças eutróficas portadoras do genótipo CC em comparação às portadoras do alelo T. Este é o primeiro estudo investigando o consumo alimentar como possível determinante dos níveis enzimáticos da PON1 em crianças. Foi verificado que aquelas que consumiram salada crua regularmente mostraram maiores níveis (p<0,01) de atividade enzimática. A associação entre o genótipo -107CC, consumo regular de salada crua, maior atividade da PON1 e concentração de HDL-C em crianças eutróficas sugere que os fatores genéticos e suas interações com o ambiente estão relacionados a um perfil protetor contra o desenvolvimento de DCV. / Childhood is a vulnerable period for the start of endothelial lesions, which may be aggravated by exposure to nutritional risk factors. On the other hand, it is proven that an elevated concentration of high density lipoprotein (HDL) protects against the emergence of these lesions, resulting in a lower risk of cardiovascular disease (CVD) development. The main antioxidant and anti-inflammatory enzyme that contributes to the antiatherogenic property attributed to HDL is paraoxonase 1 (PON1). Its serum activity may be influenced by several parameters including genetic polymorphisms, nutritional factors, lifestyle, age, drugs, health status and oxidative stress. The aim of this study was to investigate the serum PON1 activity and its association to genetic polymorphisms, biochemical and nutritional factors in children aged 5-7 years, attending pediatric clinics in two cities in southern Rio Grande do Sul. A cross-sectional study was conducted from January 2014 to February 2015. Dietary intake was assessed by SISVAN questionnaire. Weight, height and waist circumference of the child were used to evaluate nutritional status. A blood sample was collected and used to assess lipid profile and PON1 arylesterase activity, measured using phenylacetate as substrate. The biological sample was also used to determine the polymorphism PON1 C(-107)T, genotyped by PCR amplification followed by restriction digestion. Data were analyzed in Stata program version 12, assuming a 5% significance level. The median PON1 arylesterase activity was 89,4kU/L, similar between boys and girls. The polymorphism analysis revealed that the most common genotype was the heterozygous CT (52.1%). The polymorphism was in Hardy-Weinberg equilibrium. The CC genotype showed higher levels of enzyme activity than those presented by TT genotype (p<0.01). Desirable levels of HDL were related to a significant increase of the enzyme in the entire population. These lipoprotein concentrations were significantly higher only in normal weight children carrying the CC genotype compared to those carrying the allele T. This is the first study investigating food consumption as a potential determinant of PON1 enzymatic levels in children. It was verified that those who regularly consumed raw vegetables showed higher levels (p<0.01) of enzyme activity. The association between -107CC genotype, regular consumption of raw vegetables, higher PON1 activity and HDL-C concentration in normal weight children suggests that genetic factors and their interactions with the environment are related to a protective profile against the development of CVD.

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-24T13:05:15Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabrielle_Gastar.pdf: 1814034 bytes, checksum: 214de94beffb5c3d14f679611239db4b (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-24T13:56:54Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabrielle_Gastar.pdf: 1814034 bytes, checksum: 214de94beffb5c3d14f679611239db4b (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-24T13:57:01Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabrielle_Gastar.pdf: 1814034 bytes, checksum: 214de94beffb5c3d14f679611239db4b (MD5) / Made available in DSpace on 2018-05-24T13:57:01Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Gabrielle_Gastar.pdf: 1814034 bytes, checksum: 214de94beffb5c3d14f679611239db4b (MD5) Previous issue date: 2017-08-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / OBJETIVO: Estudar as associações entre os polimorfismos genéticos ECA I/D, ACTN3 R577X e PON1 C(-107)T com diabetes e hipertensão em pacientes atendidos pelo Sistema Único de Saúde. MÉTODOS: Foram coletados dados bioquímicos dos prontuários dos pacientes crônicos de três Unidades Básicas de Saúde da Família localizadas na cidade de Rio Grande. Foram aplicados questionários de consumo alimentar, nível de atividade física e socioeconômico. Foi realizada a avaliação nutricional dos pacientes e, em seguida, a coleta de saliva para a extração do DNA genômico para análise dos polimorfismos genéticos. Os polimorfismos foram analisados por PCR simples, multiplex e por polimorfismo de tamanho de fragmento de restrição. RESULTADOS: O grupo caso obteve maiores valores de peso, IMC, circunferência da cintura, percentual de gordura, pressão arterial, colesterol total, triglicerídeos e glicemia em comparação ao grupo controle. Foram encontradas associações do genótipo XX do polimorfismo R577X da ACTN3 com valores mais elevados de glicemia e triglicerídeos. O genótipo TT do polimorfismo C(-107)T da PON1 também obteve níveis maiores de triglicerídeos comparado aos outros 2 genótipos. CONCLUSÃO: O genótipo XX da ACTN3 foi fortemente relacionado com níveis baixos de HDL e altos valores de triglicerídeos, colesterol total e glicemia. Ainda encontramos altos valores de triglicerídeos e LDL no genótipo TT e menores níveis de colesterol total ligados ao genótipo CC da PON1. / OBJECTIVE: To study associations between polymorphisms in the angiotensin converting enzyme (ACE I/D), actinin 3 (ACTN3 R577X) and paraoxonase 1 (PON1 C-(107)T with chronic diseases (diabetes and hypertension) in patients attended by Sistema Único de Saúde (SUS). METHODS: Biochemical data were collected from the charts in the Basic Health Units of the Family located in the same region of the city of Rio Grande. Charts about food consumption, physical activity level and socioeconomic were applied. Nutritional data were evaluated, genomic DNA was extracted from collected saliva samples and used for analysis of genetic polymorphism. RESULTS: The case group had higher values of weight, BMI, waist circumference, body fat percentage, blood pressure, cholesterol, triglycerides and blood glucose in comparison with control group. Associations were found of XX genotype of the polymorphism R577X ACTN3 with higher values of blood glucose, and triglycerides levels. The TT genotype of the C(-107)T PON1 also had higher levels of triglycerides compared to other 2 genotypes. CONCLUSION: The genotype XX of ACTN3 was strong related with low HDL levels and high triglycerides, total cholesterol and glucose levels. Still, we realized high triglycerides and LDL leves in TT genotype of PON1 and lower leves of total cholesterol linked to CC genotype of PON1.