Submitted by (ana.araujo@ufrpe.br) on 2017-02-17T13:09:47Z No. of bitstreams: 1 Joao de Andrade Dutra Filho.pdf: 921509 bytes, checksum: f7aac66eb6fa6e2fbd3d500ad245707f (MD5) / Made available in DSpace on 2017-02-17T13:09:47Z (GMT). No. of bitstreams: 1 Joao de Andrade Dutra Filho.pdf: 921509 bytes, checksum: f7aac66eb6fa6e2fbd3d500ad245707f (MD5) Previous issue date: 2010-02-23 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The work had as aimed: (1) selecting progeny of sugarcane performancebased agronomic, index selection and genetic dissimilarity. (2) assess the genetic diversity in progenies of sugarcane by means of multivariate techniques based on eight characters agribusiness. (3) assess the genetic divergence in genotypes of sugarcane using molecular markers and microsatellites. The experiment was conducted in two stages. The first was held in the sugarcane zone of the North Coast of Pernambuco in the agricultural area of Usina Santa Teresa, Engenho Terra Rica, municipality of Goiana, with geographic coordinates (07º33’ S e 35º00’ W) and altitude of 13 m during the agricultural year 2006 to 2008 in red-yellow podzolic sandy texture. The experimental design was a randomized block design with five replications. Six progenies were evaluated, consisting of 200 individuals each, three standards considered, arising from clonal proliferation of commercial varieties RB943365, RB867515 and RB863129 and three of those self-pollinated varieties. For effective control of fertilization were used bells of TNT, measuring 0.50 cm in radius and 1.20 m long sealed. Each plot consisted of 5 rows of 8 m, spaced 1.20 m with 8 seedlings per line with 1 m between plants, thus totaling 40 seedlings per plot. The variables analyzed were: pol tons per hectare (PTH), sugarcane tons per hectere (STH), fiber (FB), correted pol% (CPP), purity (PTY), brix (BX),reducing sugar (RS), total retrievable sugar (TRS). Carried out the analysis of variance and estimation of genetic parameters, the standard Euclidean distance and the distance of Rogers were used as measures of dissimilarity matrices which were correlated by the Mantel test. The Mahalanobis distance was used to quantify the genetic divergence. Were used the method of hierarchical links averages (UPGMA) method and the optimization procedure. The second stage was performed at the Laboratory of Molecular Genetics, Department of Biology, Federal University of Lavras (UFLA) in Minas Gerais. For molecular analysis were selected 23 genotypes of the families surveyed in the first stage of the experiment. And three commercial varieties RB943365, RB867515 and RB863129 and 20 randomly selected families originating from selfing of commercial varieties(stratified mass selection in families). The genetic divergence was estimated based on the polymorphism generated by RAPD (Random Amplified Polymorphic DNA) and SSR (Simple Sequence Repeats). The arithmetic complement of the Simple Matching coefficient and Jaccard coefficient were used as measures of dissimilarity whose headquarters were correlated by the Mantel test. We applied the optimization method of Tocher and the hierarchical method of links averages (UPGMA). The methodology allowed the identification of progeny of higher genetic diversity in plant breeding by providing a safer choice of crosses to be made. / Objetivou-se com este trabalho: (1) selecionar progênies de cana-de-açúcar com base no desempenho agroindustrial, índices de seleção e dissimilaridade genética. (2) avaliar a divergência genética em progênies de cana-de-açúcar, através de técnicas multivariadas, com base em oito caracteres agroindustriais. (3) avaliar a divergência genética em genótipos de cana-de-açúcar utilizando marcadores moleculares RAPD e Microssatélites. O experimento foi conduzido em duas etapas. A primeira foi realizada na zona canavieira do Litoral Norte de Pernambuco na área agrícola da Usina Santa Tereza, Engenho Terra Rica, município de Goiana, com coordenadas geográficas (07º33’ S e 35º00’ W) e altitude de 13 m, durante o ano agrícola 2006 a 2008 em argissolo vermelho-amarelo de textura arenosa. O delineamento experimental utilizado foi o de blocos ao acaso com cinco repetições. Foram avaliadas seis progênies, constituídas de 200 indivíduos cada, sendo três consideradas padrões, oriundas de multiplicação clonal das variedades comerciais RB943365, RB867515 e RB863129 e três de autofecundação dessas mesmas variedades. Para o controle efetivo da autofecundação utilizaram-se campânulas de TNT, medindo 0,50 cm de raio e 1,20 m de comprimento totalmente fechadas. Cada parcela experimental foi constituída por 5 linhas de 8 m, espaçadas de 1,20 m com 8 seedlings por linha com 1 m entre plantas, totalizando assim 40 seedlings por parcela. As variáveis analisadas foram: Toneladas de pol por hectare (TPH), toneladas de cana por hectare (TCH), fibra (FIB), pol corrigida (PCC), pureza (PZA), teor de sólidos solúveis (BRIX), açúcares redutores (AR), açúcares totais recuperáveis (ATR). Realizou-se a análise de variância e estimativa de parâmetros genéticos, a distância euclidiana média padronizada e a distância de Rogers serviram para estimar a dissimilaridade entre progênies cujas matrizes foram correlacionadas pelo teste de Mantel. A distância generalizada de Mahalanobis foi utilizada para quantificar a divergência genética.Foram utilizados o método hierárquico de ligações médias (UPGMA) e o método de otimização de Tocher. A segunda etapa foi realizada no Laboratório de Genética Molecular do Departamento de Biologia da Universidade Federal de Lavras (UFLA) em Minas Gerais. Para a análise molecular foram selecionados 23 genótipos das famílias avaliadas na primeira etapa do experimento, sendo três variedades comerciais RB943365, RB867515 e RB863129 e 20 selecionados ao acaso das famílias oriundas da autofecundação dessas variedades comerciais (seleção massal estratificada em famílias). A divergência genética foi estimada com base no polimorfismo gerado por meio de marcadores RAPD (Random Amplified Polymorphic DNA) e SSR (Simple Sequence Repeats). O complemento aritmético do Coeficiente de Coincidência Simples e do Coeficiente de Jaccard foram utilizados como medidas de dissimilaridade, cujas matrizes foram correlacionadas pelo teste de Mantel. Foram aplicados o método de otimização de Tocher e o método hierárquico das ligações médias (UPGMA). A metodologia aplicada permitiu a identificação de progênies de maior divergência genética proporcionando aos fitomelhoristas maior segurança na escolha dos cruzamentos a serem realizados.

Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / A anemia falciforme (AF) à uma hemoglobinopatia hereditÃria autossÃmica causada por uma mutaÃÃo pontual no gene da beta globina gerando uma hemoglobina anormal denominada de hemoglobina S (HbS), em homozigose. A doenÃa se caracteriza por apresentar uma variabilidade do quadro clÃnico, que se deve à mÃltiplos fatores, dentre eles a concentraÃÃo de hemoglobina fetal (HbF), os haplÃtipos do gene da beta globina e os polimorfismos do gene BCL11A, entre outros. A avaliaÃÃo dos moduladores genÃticos na AF tem sido desenvolvida com a finalidade de melhorar o entendimento da sua fisiopatologia e direcionar a abordagem terapÃutica objetivando sua individualizaÃÃo. A pesquisa se propÃs a determinar a modulaÃÃo genÃtica dos polimorfismos do gene BCL11A (rs4671393, rs7557939 e rs1186868) sobre o perfil inflamatÃrio, hemolÃtico, no estresse oxidativo e nas concentraÃÃes das HbF, HbS nos pacientes portadores de AF, em estado estacionÃrio. O estudo foi do tipo transversal e analÃtico com 42 pacientes adultos, em acompanhamento ambulatorial no Hospital UniversitÃrio Walter CantÃdio (HUWC), com diagnÃstico molecular e haplÃtipos do gene da beta globina S previamente realizados. Os pacientes estavam em uso de HidroxiurÃia (HU), em mÃdia, 20mg/kg de peso corporal. Amostras biolÃgicas de sangue perifÃrico foram obtidas para a realizaÃÃo dos exames laboratoriais: as dosagens das citocinas prà inflamatÃrias IL-6, IL-17, TNF-alpha e das antiinflamatÃrias IL-10 e TGF-beta, por Elisa; contagem de reticulÃcitos por metodologia manual, dosagem de metemoglobina (MetHb) e lactato desidrogenase (LDH), por espectrofotometria; do nitrito (NOx), malonaldeÃdo (MDA) sÃricos, as enzimas antioxidantes eritrocitÃrias, catalase (CAT) e da glutationa peroxidase (GPx) por kits e espectrofotometria. Os polimorfismos genÃticos do gene BCL11A nas regiÃes, rs4671393, rs7557939 e rs1186868 foram determinados por Real Time PCR. As dosagens da HbF e HbS foram realizadas por HPLC (High Performance Liquid Chromatography). Os dados idade, sexo e eventos clÃnicos foram obtidos dos prontuÃrios. Toda a anÃlise estatÃstica foi realizada usando o software livre R, na versÃo 3.1.2. Para anÃlise da frequÃncia do sexo e dos genÃtipos, por regiÃo e das associaÃÃes entre o tipo de haplÃtipo e dos eventos clÃnicos com as regiÃes do BCL11A, foram usados os testes de Qui-quadrado e o exato de Fisher. Realizou-se o teste paramÃtrico de ANOVA (obtido sob suposiÃÃes distribucionais), bem como o teste nÃo-paramÃtrico de Kruskal-Wallis para a anÃlise da associaÃÃo dos genÃtipos do gene BCL11A com a idade, os nÃveis de HbS, HbF, perfil inflamatÃrio, hemolÃtico e do estresse oxidativo. Foi considerado significante ao nÃvel de 5%. A maioria dos pacientes (57,14%) era do sexo feminino. A idade dos pacientes incluÃdos foi de 18 a 65 anos, com valor mÃdio e mediano de 35,1 e 33 anos, respectivamente. Somente a rs7557939 do BCL11A, o genÃtipo A/G foi o mais prevalente e a prevalÃncia do genÃtipo A/G foi maior nas mulheres , enquanto nos homens a prevalÃncia maior foi do genÃtipo A/A. No entanto, a rs1186868 do BCL11A, a maioria (56,52%) das mulheres apresentaram o genÃtipo C/T e a metade dos homens apresentaram o genÃtipo T/T. Nenhuma regiÃo do gene BCL11A apresentou associaÃÃo significativa com os haplÃtipos do gene da beta globina S. Em relaÃÃo a moduÃÃo do gene BCL11A com os nÃveis de HbS e HbF, verificou-se que na rs1186868 houve resultado significativo do genÃtipo mutante T/T, que apresentou maiores nÃveis de HbS e menores nÃveis de HbF. Na rs7557939 houve uma diminuiÃÃo significante de HbF no alelo mutante A/A, porÃm, nÃo houve relaÃÃo com a HbS. NÃo houve associaÃÃo entre os SNPs, nas trÃs regiÃes estudadas, com relaÃÃo ao nÃmero mÃdio/mediano dos moduladores inflamatÃrios, marcadores de hemÃlise, do estresse oxidativo e dos eventos clÃnicos, ao nÃvel de 5%.Os achados reforÃam a hipÃtese da moduÃÃo genÃtica dos polimorfismos do gene BCL11A em relaÃÃo aos nÃveis de HbF, onde os alelos selvagens, nas regiÃes rs7557939 e rs1186868 apresentaram um carÃter protetor no prognÃstico em decorÃncia de terem apresentado aumento dos nÃveis de HbF, nos pacientes com AF do estudo.

Submitted by Erika Demachki (erikademachki@gmail.com) on 2015-02-24T16:18:57Z No. of bitstreams: 2 Dissertação - Silene Jacinto da Silva - 2012.pdf: 1244943 bytes, checksum: 169827dfbf3ff42a25f37a6dbdd252f2 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Erika Demachki (erikademachki@gmail.com) on 2015-02-24T16:21:26Z (GMT) No. of bitstreams: 2 Dissertação - Silene Jacinto da Silva - 2012.pdf: 1244943 bytes, checksum: 169827dfbf3ff42a25f37a6dbdd252f2 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2015-02-24T16:21:26Z (GMT). No. of bitstreams: 2 Dissertação - Silene Jacinto da Silva - 2012.pdf: 1244943 bytes, checksum: 169827dfbf3ff42a25f37a6dbdd252f2 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2012-04-23 / The Deletion/Insertion polymorphisms of the angiotensin converting enzyme genes and the A1166C of the angiotensin II receptor AT1R were analyzed in a cohort of 90 patients, among 30 carriers of chronic heart failure, aging from 30 to 86, in which 66, 6% were males. The control group was based on 60 cardiopathic patients without heart failure matched by age and gender. The heart failure was attributed to the etiologies: chagas cardiomyopathy (46,7%), idiopathic dilated cardiomyopathy and others (23,3%), hypertensive cardiomyopathy (20%) and ischemic cardiomyopathy (10%). In order to determine the genotypes the PCR - RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) technique was applied. The genotypes distribution was analyzed, as well as the allele’s frequency and the possible polymorphisms associations with different clinical variations and heart failure carrier’s evolution during 12 months. For the analysis of descriptive and inferential statistical were used the t test, chi-square (χ2) test, Kaplan - Meier and ANOVA. The distribution of the genotypes D/I of the genes ACE and the polymorphisms A1166C of the angiotensin II was similar between the two groups (p = 0,23 e p= 0,12). The evaluation of the polymorphisms studies presented a lack of association with the clinical variations analyzed and the evolution of the heart failure carriers during 12 months. / Os polimorfismos de Deleção/Inserção do gene da enzima conversora da angiotensina e A1166C do receptor AT1R da angiotensina II foram estudados em uma coorte com 90 pacientes, sendo 30 portadores de insuficiência cardíaca crônica, com idades variando entre 30 e 86 anos, dos quais 66,6% eram do sexo masculino. O grupo controle foi constituído por 60 pacientes cardiopatas, porém sem insuficiência cardíaca, pareados por idade e sexo. A insuficiência cardíaca foi atribuída às seguintes etiologias: cardiomiopatia chagásica (46,7%), cardiomiopatia dilatada idiopática e outras (23,3%), cardiomiopatia hipertensiva (20%) e cardiomiopatia isquêmica (10%). A determinação dos genótipos foi realizada pelas técnicas PCR - RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism). Foram avaliadas as distribuições dos genótipos, as frequências alélicas, as prováveis associações dos polimorfismos com diferentes variáveis clínicas e a evolução dos portadores de insuficiência cardíaca no seguimento de 12 meses. Para a análise da estatística descritiva e inferencial utilizou-se o teste t, teste Qui-quadrado (χ2), kaplan - Meier e ANOVA. A distribuição dos genótipos D/I do gene ECA e do polimorfismo A1166C foi semelhante entre os dois grupos (p=0,23 e p=0,12). A avaliação dos polimorfismos estudados apontou para a ausência de associação com as variáveis clínicas analisadas e com a evolução dos portadores de insuficiência cardíaca durante o seguimento de 12 meses.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O cÃncer gÃstrico apresenta, mundialmente, uma elevada taxa de mortalidade, com alta incidÃncia no Brasil, sendo a infecÃÃo com Helicobacter pylori um fator de risco bem estabelecido. Dependendo da presenÃa de genes de virulÃncia como cagA, cagE, vacA e virB11, H. pylori pode causar respostas inflamatÃrias diferenciadas, apresentando grande quantidade de Ãxido nÃtrico (ON) gerado principalmente por iNOS. Quantidade elevada de ON resulta em acÃmulo de espÃcies reativas do oxigÃnio cuja instabilidade causa danos oxidativos no DNA. A integridade genÃmica à garantida por enzimas de reparo importantes como: APE-1, OGG-1, e PARP-1. Polimorfismos genÃticos que modifiquem a atividade dessas enzimas podem influenciar a capacidade de reparo e, portanto, a susceptibilidade do hospedeiro ao desenvolvimento do cÃncer gÃstrico associado à H. pylori. Assim o objetivo deste estudo foi avaliar a associaÃÃo dos polimorfismos C150T em iNOS, T2197G em APE-1, C1245G em OGG-1 e A40676G em PARP-1 com o genÃtipo de H. pylori em 109 amostra de pacientes diagnosticados com adenocarcinomas gÃstricos atendidos em hospitais de Fortaleza, CearÃ. A identificaÃÃo dos polimorfismos foi feita por PCR-RFLP e a detecÃÃo e genotipagem de H. pylori foram feitas por PCR. Os polimorfismos estudados apresentaram as seguintes freqÃÃncias: iNOS - 78% CC, 21,1% CT e 0,9% TT; PARP-1- 69,7% AA, 26,6% AG e 3,7% GG, para OGG-1 56% CC, 39,4% CG, e 4,6% GG e para APE-1 38,5%TT, 47,7%TG e 13,8% GG. Salienta-se a baixa freqÃÃncia do genÃtipo polimÃrfico (TT) de iNOS e alta frequÃncia do heterozigoto (TG) de APE. Os alelos variantes de iNOS e de PARP-1 foram correlacionadas com indivÃduos ≤55 anos, sugerindo que estes polimorfismos estariam associados ao desenvolvimento precoce da neoplasia. Os tumores intestinais localizados na regiÃo nÃo-antro correlacionaram-se com o genÃtipo OGG-1 CG; enquanto que os difusos, localizados no corpo com o genÃtipo AA de PARP-1. H. pylori foi detectada em 92,6% dos casos. Os genes cagA, cagE e virB11 foram detectados em 65,3%, 50,4% e 60,3% dos casos, respectivamente e vacAs1m1 detectado em 72,2%. Os casos foram agrupados considerando os alelos de vacA e a integridade da ilha de patogenicidade cag, sendo os grupos A1 e A2, composto por cepas mais patogÃnicas, o qual foi observado em 33,6% e 13,8% dos pacientes, respectivamente. Na anÃlise individual de cada enzima, observou-se que os indivÃduos portadores dos alelos variantes de APE-1 (TG+GG) estavam infectados com cepas pouco patogÃnicas (p=0,0422). Essas cepas pouco patogÃnicas tambÃm foram associadas aos pacientes portadores do genÃtipo selvagem (AA) de PARP-1 (p=0,0396). Esses dados foram confirmados quando os pacientes infectados por cepas mais virulentas foram comparadas aos infectados por cepas menos virulentas (p=0,046). Analisando apenas o grupo A1 observou-se tambÃm uma correlaÃÃo de APE-1 (TG) com OGG-1(CC). Quando os genÃtipos foram combinados considerando somente as enzimas de reparo estudadas ou duas a duas, verificou-se que parte dos pacientes infectados com o genÃtipo selvagem de PARP-1 eram portadores do alelo variante para pelo menos uma das enzimas e que parte dos pacientes infectados com cepas menos patogÃnicas possuÃam o alelo polimÃrfico de APE-1, independente do genÃtipo da enzima de reparo associada. Somados, esses dados indicam a relevÃncia do polimorfismo da APE-1 no desenvolvimento do cÃncer gÃstrico em indivÃduos infectados com cepas menos virulentas e corroboram com a importÃncia do genÃtipo bacteriano, uma vez que, de maneira geral, indivÃduos com genÃtipo selvagem para as enzimas de reparo estudadas desenvolveram cÃncer gÃstrico quando infectados por cepas virulentas. / Gastric cancer is the most deadly malignant neoplasia worldwide, with high incidence in Brazil and Helicobacter pylori infection is a well-established risk factor. Depending on the presence of virulence genes such as cagA, cagE, vacA and virB11, H. pylori can cause differentiated inflammatory responses, with large amounts of nitric oxide (NO) generated primarily by iNOS. High amount of NO resulting in accumulation of reactive oxygen species can cause DNA oxidative damage. The genomic integrity is guaranteed by important repair enzymes as: APE-1, OGG-1 and PARP-1. Genetic polymorphisms that modify the activity of these enzymes may influence the ability to repair and therefore the host susceptibility to the development of gastric cancer H.pylori associated. Therefore, the goal of this study was to evaluate the association of the C150T polymorphism in iNOS, T2197G in APE-1, C1245G in OGG -1 and A40676G in PARP-1 with H.pylori genotype in 109 cases of patients with gastric adenocarcinoma from hospitals in Fortaleza, CearÃ. The identification of polymorphisms was performed by PCR-RFLP and the detection and genotyping of H.pylori were performed by PCR. The studied polymorphisms showed the following frequencies: iNOS 78% CC, 21.1% CT and 0.9% TT; PARP-1 69.7% AA 26.6% AG and 3.7% GG to OGG -1 56% CC, 39.4% CG and 4.6% GG and APE-1 38.5% TT, 47.7% TG and 13.8% GG. Valuable to note the low frequency of the homozygous polymorphic (TT) of iNOS and the high frequency of heterozygous (TG) from APE-1. The variant alleles of iNOS and PARP-1 were correlated with subjects ≤ 55 years, suggesting that these polymorphisms were associated with early development of the neoplasia. Intestinal tumors located in the non-antrum were correlated with heterozygous genotype of OGG-1 (CG), while diffuse, located on the body with the AA genotype of PARP-1. H. pylori was detected in 92.6% of cases. The genes cagA, cagE and virB11 were detected in 65.3%, 50.4% and 60.3% of cases respectively and vacAs1m1 was detected in 72.2%. The cases were also grouped considering the alleles of vacA and the integrity of the cag-pathogenicity island. Thus, the groups A1 and A2, consist of more pathogenic strains, were observed in 33.6% and 13.8% of patients, respectively. In the individual analysis of each enzyme, we observed that individuals carrying the variant alleles of APE-1 (TG+GG) were infected with low pathogenic strains (p=0.0422). These low pathogenic strains were also associated with patients carrying the wild genotype (AA) of PARP-1 (p=0.0396). These data were confirmed when patients infected with more virulent strains were compared to those infected with less virulent strains (p = 0.046). Analyzing only the group A1, it was also observed a correlation of APE-1 (TG) with OGG-1 (CC). When genotypes were combined by considering only the repair enzymes studied or two by two, it was found that most patients infected with the wild-type of PARP-1 were carriers of the variant allele for at least one of the enzymes and that most patients infected with less pathogenic strains possess a polymorphic allele of APE-1, independent of the genotype associated with the repair enzyme. Taken together, these data indicate the relevance of the APE-1 polymorphism in the development of gastric cancer in individuals infected with less virulent strains and corroborate the importance of the bacterial genotype, since; in general, individuals with wild-type for enzymes repair studied developed gastric cancer when infected with virulent strains.

Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2014-09-19T12:13:58Z No. of bitstreams: 2 Dissertacao Lucas C G Pereira.pdf: 869540 bytes, checksum: 9c40d119515c83b8b52606c52be6703d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-09-19T12:56:29Z (GMT) No. of bitstreams: 2 Dissertacao Lucas C G Pereira.pdf: 869540 bytes, checksum: 9c40d119515c83b8b52606c52be6703d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-09-19T12:56:29Z (GMT). No. of bitstreams: 2 Dissertacao Lucas C G Pereira.pdf: 869540 bytes, checksum: 9c40d119515c83b8b52606c52be6703d (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2013-05-02 / In recent years, the evolution of health expenditures and specifically drugs, has worried governments. Among the various specialties, oncology is among those dealing with the greatest difficulties in the management of drug therapy. It is known that patients treated with various drugs have variability of response and susceptibility to drug toxicity. In present work, we study the role of Multiple Drug Resistance gene (MDR1) polymorphisms C1236T and C3435T frequencies and response to treatment with imatinib mesylate in 96 patients with chronic myeloid leukemia (CML). A total of 96 patients with CML were treated according to the Brazilian National Cancer Institute (INCA) guidelines and the blood samples were collected for genotyping. Genomic DNA was extracted and C1236T and C3435T polymorphisms genotyping was performed by the polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP), which detects a variation in length of a DNA fragment generated (370pb and 340pb) by a specific endonuclease in a specific site of the genome (HaeIII and MboI). Of the 96 CML samples, 31 samples were homozygous (CC), 13 homozygous (TT) and 52 heterozygous (CT) for exon 12 (1236). For the exon 26 (3435), 35 were homozygous (CC), 12 homozygous (TT) and 49 heterozygotes (CT). All frequencies for both polymorphisms were in Hardy-Weinberg equilibrium (p = 0.229 and q = 0.414). We found percentage association between polymorphisms and their distribution in different populations, and the response to treatment both cytogenetic and molecular difference was not statistically significant (p <0.05) when compared to age and sex presented response by patients and also no statistical difference (p <0,050). We conclude that the observed allele frequency for exons 1236 were 59.4% for C and 40.6% for T and the frequencies for the exon 3435 were 62.0% for C and 38.0% for T. That the relationship between the frequencies of polymorphisms of MDR1 in populations of different geographic locations, can provide tools that help in choosing a more appropriate and effective treatment of CML. / Neste estudo, o papel dos polimorfismos C1236T e C3435T do gene de Resistência a Múltiplas Drogas (MDR1) foram investigados em relação à frequência e a resposta ao tratamento com imatinibe em pacientes com leucemia mielóide crônica (LMC). Um total de 96 pacientes com LMC foram tratados de acordo com as diretrizes do Instituto Nacional do Câncer (INCA) e amostras de sangue foram coletadas para genotipagem do gene MDR (Resistência à Multiplas Drogas). O DNA genômico foi extraído e a genotipagem dos polimorfismos C1236T e C3435T foi realizada por meio da reação em cadeia da polimerase com fragmentos de restrição (PCR-RFLP), que detectou uma variação no comprimento de um fragmento de DNA gerado (370pb e 340pb) por uma endonuclease específica em um sítio específico do genoma (HaeIII e Mbol). Analisando as 96 amostras de pacientes para o polimorfismo no éxon 12 (1236) com LMC, 31 amostras apresentaram homozigose (CC), 13 homozigose (TT) e 52 heterozigose (CT). Para o estudo do polimorfismo no éxon 26 (3435), 35 foram homozigotas (CC), 12 homozigotas (TT) e 49 heterozigotas (CT). Todas as frequências para ambos os polimorfismos apresentaram-se em equilíbrio de Hardy-Weinberg (p = 0,229 e q = 0,414). Foi encontrada associação do percentual dos polimorfismos estudados em relação à distribuição dos mesmos em grupos de diferentes localizações geográficas, e sobre a resposta ao tratamento tanto citogenética e molecular, não houve diferença estatísticamente significante (p<0,05), quando foi comparado à idade e ao gênero apresentados pelos pacientes e a resposta também não houve diferença estatística (p<0,05). Conclui-se que as frequências alélicas observadas para o éxon 1236 foram de 59,4% para C e 40,6% para T e as frequências para o éxon 3435 foram de 62,0% para C e 38,0% para T e que a relação entre as frequências de polimorfismos de MDR1 nas populações de diferentes localizações geográficas, pode fornecer ferramentas que auxiliem na escolha de um tratamento mais adequado e eficaz da LMC.

Este trabalho investiga a influência de polimorfismos na região promotora do gene das metaloproteases 1, 3 e 8 na fisiopatogenia da insuficiência primária do tendão tibial posterior. A amostra de 150 pacientes selecionados é dividida em grupo-teste: 50 pacientes com diagnóstico clínico e anatomopatológico de tendinopatia do tendão tibial posterior e grupo-controle: 100 pacientes com tendão tibial posterior íntegro. O DNA dos voluntários é obtido a partir de células epiteliais da mucosa bucal mediante a extração com acetato de amônia. As técnicas de PCR e RFLP são utilizadas para análise dos genótipos. A análise estatística dos resultados é realizada pelo teste do qui-quadrado com nível de significância de 5%. Os resultados mostram que os polimorfismos -1607 da MMP-1 e -799 da MMP-8 estão relacionados com risco maior para tendinopatia primária do tendão tibial posterior, enquanto o polimorfismo -1612 da MMP-3 parece não influenciar essa tendinopatia / The aim of this study was to investigate the influence of polymorphisms in the promoter region of the gene of metalloproteinases 1, 3 and 8 in physiopatology of primary posterior tibial tendon insufficiency. The sample of 150 selected patients was divided into test group: 50 patients undergoing surgical procedures and pathological diagnosis of degenerative lesions of the posterior tibial tendon, and control group: 100 patients with posterior tibial tendon intact and no signs of degeneration. The DNA of the volunteers was obtained from oral mucosa epithelial cells, by extraction with ammonium acetate. PCR and RFLP were used for analysis of genotypes. Statistical analysis of results was performed by Chi-squared test with significance level of 5%. The results show that polymorphisms -1607 of MMP-1 and -799 of the MMP-8 are associated with increased risk for primary tendinopathy of the posterior tibial tendon, while the -1612 polymorphism of MMP-3 does not influence this tendinopathy

As variantes genéticas LEP G-2548A, LEP 3\'HVR, D1S200 (LEPR),D18S858 (MC4R) e D2S1788 (POMC)foram avaliadas em 100 indivíduos obesos (GE) e 110 não-obesos (GC) brancos. A genotipagem desses indivíduos foi realizada por PCR e RFLP. As freqüências dos alelos LEP -2548G e LEP 3\'HVR-Classe I no grupo GE foram maiores que no GC P<0,05). O haplótipo LEP G/I foi mais freqüente no grupo GE (P=0,018) e nos indivíduos com obesidade central (P=0,047). As freqüências dos alelos 41/42 da D18S858 e do alelo 17 da D1S200 foram maiores (P<0,05) no grupo GE. Os indivíduos com alelos LEP 3\'HVR-Classe I, 41/42 da D18S858 e 17 da D1S200 apresentaram valores mais altos de índice de massa corporal (IMC) e circunferência abdominal (P<0,05). Em conclusão, as variantes LEP G-2548A, LEP 3\'HVR, D18S858 e D1S200 estão associadas com a obesidade e com variações nos valores de IMC e circunferência abdominal em indivíduos brasileiros brancos. / The genetic variants LEP G-2548A, LEP 3\'HVR, D1S200 (LEPR), D18S858 (MC4R) D2S1788 (POMC) were studied in groups of 100 obese (GE) and 110 non-obese (GC) white individuals. Genotyping were carried out by PCR and RFLP techniques. Alleles frequencies from LEP -2548G and Class I alleles from LEP 3\'HVR were higher in GE group, when compared to GC group (P<0,05). The LEP G/I haplotype was more frequent in GE group (P=0,018) and in individuals with central obesity (P=0,047). Alleles 41/42 from D18S858 and allele 17 from D1S200 were more frequent (P<0,05) in GE group. Individuals carrying LEP 3\'HVR-Class I, alleles 41/42 from D18S858 and allele 17 from have shown elevated body mass index (BMI) and waist circumference values (P<0,05). In conclusion, the LEP G-2548A, LEP 3\' HVR, D1S200, and D18S858 genetic variants were found to be associated to obesity and variations in BMI and waist circumference in Brazilian white individuals.

In this study, the role of two DNA repair genes, X-ray repair cross complementing group 3 (XRCC3) Thr241Met and Rad51 G135C polymorphisms were investigated in the risk of development of childhood ALL in Turkish population among 193 healthy controls and 184 ALL patients, by using PCR-RFLP technique. For XRCC3 Thr241Met polymorphism, the frequencies of both heterozygous and homozygous mutant genotypes were found to be higher in the controls compared to ALL patients (OR: 0.59, p = 0.02 / OR: 0.48, p = 0.02, respectively). In addition, either heterozygous (Thr/Met) or homozygous mutant (Met/Met) genotypes were significantly more common in the controls than the ALL patients (OR: 0.55, p =0.005). In case of Rad51 G135C polymorphism, no significant associations have been found with the risk of childhood ALL. Combination of XRCC3 heterozygote and Rad51 heterozygote genotypes increased the protective effect for risk of childhood ALL. (OR=0.35 / p =0.02). Combination of homozygote mutant genotype of XRCC3 with homozygote wild type genotype of Rad51 gave a highly statistically proved protective effect for the development of disease (OR= 0.36 / p= 0.004). To our knowledge, this is the first study showing the protective role of XRCC3 Thr241Met polymorphism either alone or in combination with Rad51 G135C variant on the risk of development of childhood ALL. In addition, interactions of these polymorphisms with non-genetic risk factors were investigated. Only in terms of paternal exposure, the heterozygote (Thr/Met) genotype for XRCC3 gene in children whose father exposed to cigarette smoke demonstrated a significant risk of 3.0 fold (p=0.05). Moreover, the frequency of Rad51 135C allele was determined for the first time in Turkish population. The frequency of the mutant allele was found to be very similar to that observed in other Caucasian populations.

ANALYSIS OF CYTOCHROME P4501A1 GENETIC POLYMORPHISMS IN PATIENTS WITH ISCHEMIC STROKE Adali, Ayse &Ccedil / inar M.Sc., Department of Biochemistry Supervisor: Prof. Dr. Orhan Adali Co-Supervisor: Dr. Birsen Can Demird&ouml / gen January 2011, 179 pages Stroke is the third leading cause of death worldwide and results in serious disabilities. Cytochrome P450 1A1 gene (CYP1A1) is a highly polymorphic gene encoding its corresponding xenobiotic metabolizing enzyme which is responsible from the metabolism of carcinogenic polycyclic aromatic hydrocarbons (PAHs) that are engaged with the formation of free radicals. Atherosclerosis is a major cause of ischemic stroke and this pathology may be associated with the disruption of vascular homeostasis due to the formation of these chemicals. The main objective of this study was to investigate the coding region (A4889G) and non-coding region (T6235C) polymorphisms of the CYP1A1 gene as a risk factor for ischemic stroke. The study group in Turkish population consisted of 226 unrelated ischemic stroke patients and 113 control subjects. There was no statistically significant difference between the groups with respect to age and gender. Total blood samples were obtained from G&uuml / lhane Military Medical Academy Hospital, Neurology Department, Ankara. In stroke patients, hypertension, diabetes mellitus, smoking and obesity were at least 2 times more common and high density lipoprotein cholesterol (HDL-C) was significantly lower than controls. The frequency of mutant allele 4889G was 0.445 in patients and was nearly the same with controls. The frequency of mutant allele 6235C was 0.151 in patients and was significantly higher in controls (0.226, P=0.015). The risk of diabetic, smoker and obese individuals having ischemic stroke was significantly higher in 4889G allele carriers (AG+GG / Odds ratio / OR= 2.1, 2.4 and 3, respectively). The risk of hypertensive and diabetic individuals having ischemic stroke was higher in 6235TT genotypic people (OR= 3 and 2.2, respectively). On the contrary, the risk of smoker and obese individuals having ischemic stroke was significantly higher in 6235 C allele carriers (OR=5.3 and 3.7, respectively). Logistic regression analysis revealed that hypertension, smoking, levels of low density lipoprotein cholesterol (LDL-C) and HDL-C and 6235C allele were significant predictors of stroke. In this analysis, high level of LDL-C was found to be associated with almost 1.5-fold risk of ischemic stroke. On the other hand, HDL-C and having mutant 6235C allele decreased the risk of ischemic stroke 2.5 and 2-fold, respectively. This is the first study investigating the relation between A4889G polymorphism and stroke risk. Additionally, in Turkish population A4889G and T6235C polymorphisms were analyzed for the first time in terms of its relation to ischemic stroke. The present study demonstrated that the frequency of mutant 4889G allele was nearly the same in stroke patients and control subjects / whereas the frequency of mutant 6235 C allele was higher in control subjects than in stroke patients. Consequently, we decided that carrying mutant 4889 G allele does not constitute a risk for ischemic stroke and carrying mutant 6235C allele may have a protective effect against stroke.

Interleukin-1 (IL-1) is a prototypic multifunctional cytokine. IL-1 family include interleukin-1 a (IL-1 a), interleukin-1b (IL-1 b) and interleukin-1 receptor antagonist (IL-1 Ra). IL-1 b is the archetypeal pleiotropic cytokine which have been produced by many cells and exerting its biological effects on almost all cell types. IL-1 b is the most potent of known agents that are gastric cytoprotective, antiulcer, antisecretory and an inhibitor of gastric emptying. IL-1 Ra competes with IL-1 b for cell surface receptor occupancy. Host genetic factors that affect interleukin-1 (IL-1) have been reported to influence the susceptibility of Caucasians to gastric cancer. Whether Asians have the same genetic susceptibility remains unclear. In this study, the genetic associations of IL-1B and IL-1RN polymorphisms with gastric cancer and duodenal ulcer in Taiwan were evaluated. Genomic DNA from 140 unrelated Taiwanese patients with gastric adenocarcinoma, 94 with duodenal ulcer and 165 ethically matched healthy controls was typed for polymorphisms at positions ¡V31, -511, and +3954 in the IL-1B gene, and the variable number of tandem repeats polymorphisms in intron 2 of the IL-1RN gene. The allele frequencies of IL-1RN 2R in gastric cancer cases were much higher than those in healthy controls (9% vs. 3%, p = 0.781). The allele frequencies of IL-1B ¡V31, IL-1B ¡V511 and IL-1B +3954 did not differ. An increased risk of the development of intestinal type gastric carcinoma was found in IL-1RN 2R carriers with an odds ratio (OR) of 4.06 (95% confidence interval [CI]: 1.68 ¡V 9.79, p-value=0.085). And another increased risk of the development of diffuse type gastric carcinoma was found in IL-1RN 2R carriers with an odds ratio (OR) of 3.15 (95% confidence interval [CI]: 1.16 ¡V 8.56, p-value=0.061). A significant association was found in IL-1RN 2R/4R genotype and the risk of the development of duodenal ulcer, with an odds ratio (OR) of 2.57 (95% CI: 1.03 ¡V 6.38, p = 0.292). No significant relationship was noted in duodenal ulcer patients with IL-1B genotype examed in this study. Additionally, a synergistic interaction between blood type A and IL-1 RN 2R carriers existed in gastric cancer patients (OR= 4.51; 95% CI: 1.20 ¡V 16.88, p-value=0.516). The synergistic interaction was even stronger between blood type O and IL-1 RN 2R carriers of duodenal ulcer patients (OR= 10.3; 95% CI: 2.10 ¡V 50.61, p-value=0.160). In conclusion, the genetic polymorphisms of IL-1RN 2R and blood type A are associated with the development of gastric cancer. The genetic polymorphisms of IL-1RN 2R and blood type O are associated with the development of duodenal ulcer.