FundaÃÃo de Amparo à Pesquisa do Estado do Cearà / O H. pylori infecta atualmente metade da populaÃÃo mundial e à relacionado com o desenvolvimento das afecÃÃes gÃstricas. O Enteroteste à um mÃtodo minimamente invasivo que pode ser usado para detecÃÃo do H. pylori por meio nÃo endoscÃpico. O objetivo deste estudo foi avaliar a sensibilidade e especificidade do Enteroteste frente ao Teste RespiratÃrio e estudar o perfil genÃtico das cepas de H. pylori em indivÃduos residentes na comunidade Parque UniversitÃrio. O suco gÃstrico foi colhido a partir do Enteroteste, e realizada a cultura e a extraÃÃo do DNA do H. pylori. A genotipagem das cepas foi realizada atravÃs da tÃcnica de PCR e os sÃtios de fosforilaÃÃo EPIYA de sequenciamento Participaram do estudo 50 indivÃduos, 43 positivos e 7 negativos para a infecÃÃo pelo H. pylori atravÃs do Teste RespiratÃrio. A cultura e o PCR a partir do Enteroteste apresentaram sensibilidade de 86% e 77%, respectivamente, com especificidade de 100% em ambos os mÃtodos 33 cepas foram genotipadas como vacA positivas, sendo 39,4% vacA s1m1; 15,2% vacA s1m2; 18,2% vacA s2m2 27,2% com perfil vacA s com ausÃncia do alelo m. Foram cagA positivas 66,6% das cepas sendo 54,5% com perfil EPIYA-ABC, 41,0% EPIYA-ABCC e 4,5% EPIYA-AB. O Enteroteste mostrou-se um mÃtodo confiÃvel com boa sensibilidade e especificidade para identificaÃÃo do H. pylori atravÃs do cultivo e da tÃcnica de PCR. A maioria das cepas expressou o alelo vacA s1, com predomÃnio do subtipo s1b e da combinaÃÃo alÃlica s1m1. Mais da metade das cepas estudadas expressaram o gene cagA e a maioria dessas cepas tinham 3 ou 4 sÃtios de fosforilaÃÃo, com perfil EPIYA-ABC ou EPIYA-ABCC. O perfil genÃtico apresentado por essas cepas à o descrito para a AmÃrica do Sul e diferente do padrÃo AsiÃtico, a maioria das cepas circulantes na comunidade tÃm importante potencial patogÃnico. O Enteroteste à um mÃtodo seguro, sensÃvel e especÃfico para detecÃÃo do H. pylori. / The H. pylori infect currently half of the worldâs population and is related to the development of gastric disorders. The Enterotest is a minimally invasive method that can be used for detection of H. pylori through endoscopic not. The objective of this study was to evaluate the sensitivity and specificity of Enterotest opposite the Respiratory Testing and studying the genetic profile of strains of H. pylori individuals resident in the Community College Park. The gastric juice was collected from Enteroteste, and held the culture and the extraction of DNA from H. pylori. The genotyping of strains was carried out by the PCR technique and phosphorylation sites EPIYA sequencing participated in the study 50 individuals, positive and negative 7 43 for infection H. pylori through Respiratory Test. Culture and PCR from Enteroteste showed sensitivity of 86% and 77%, respectively, with 100% specificity in both methods. 33 strains were positive, genotipadas as Cow being 39.4% Cow s1m1; 15.2% Cow s1m2; 18.2% Cow s2m2 27.2% with Cow s profile with absence of allele m. Were positive 66.6% Bran of strains being profiled 54.5% EPIYA-ABC, 41.0% EPIYA-ABCC and 4.5% EPIYA-AB. The Enterotest proved to be a reliable method with great sensitivity and specificity for identification of H. pylori through cultivation and PCR technique. Most strains expressed alelo Cow s1, with a predominance of subtype s1b and allele combination s1m1. More than half of the studied strains expressed gene Shit and most of these strains were 3 or 4, phosphorylation sites profiled EPIYA-ABC or EPIYA-ABCC. The genetic profile presented by these strains is described for South America and nonstandard Asia, most strains circulating in the community have important potential pathogenic. The Enterotest is a safe method, sensitive and specific detection of H. pylori.

Helicobacter pylori est associée à des pathologies gastro-duodénales sévères mais est également un marqueur bactérien de migrations humaines. Nous avons montré que des populations génétiques distinctes de H. pylori ont accompagné au moins quatre migrations en Asie du sud-est et en Océanie : i) une expansion des ancêtres des austronésiens il y a 5000 ans à partir de Taiwan en Océanie, ii) une migration d’Inde en Asie du sud-est depuis 2000 ans,iii) une migration des ancêtres des locuteurs des langues austro-asiatiques au Vietnam et auCambodge il y a 4000 ans, i) une migration des ancêtres des Thaïs du sud de la Chine vers l’actuelle Thaïlande au début du second millénaire. Ces données confirment la résolution plus élevée de la diversité génétique de H. pylori pour retracer les anciennes migrations humaines par comparaison aux marqueurs génétiques humains traditionnels. Nous avons ensuite investigué les facteurs de virulence de souches isolées de patients présentant des symptômes gastriques au Sénégal et au Cambodge. Au Sénégal, une association significative a été observée entre le cancer gastrique et le gène cagA, deux motifs EPIYA-C et l’allèle vacA s1. De multiples segments EPIYA-C étaient observés moins fréquemment que dans les autres régions du monde, contribuant probablement à la faible incidence du cancer gastrique. Au Cambodge, une introgression fréquente d’allèles cagA et vacA européens dans des souches d’Asie de l’est a été observée. CagA et VacA ayant des effets antagonistes, cette expansion pourrait entraîner la rupture de l’équilibre entre les effets biologiques de ces deux protéines et être responsable de conséquences graves sur l’évolution de la maladie. / Helicobacter pylori is associated with severe gastroduodenal disorders but is also a bacterial genetic marker of human migrations. First, we provide evidence that distinct H. pylori genetic populations accompanied at least four ancient human migrations into Oceania and Southeast Asia: i) an expansion of Austronesian speaking people about 5000 years ago from Taiwan into Oceania, ii) a migration from India into Southeast Asia within the last 2000 years, iii) a migration of Austro-Asiatic speaking people into Vietnam and Cambodia about 4000 years ago, and iv) a migration of the ancestors of the Thai people from Southern China into Thailand during the early second millennium AD. These findings demonstrate that H. pylori genetic diversity has more discriminatory power than traditional human genetic markers for tracing old human migrations. Second, we investigated virulence factors of H. pylori strains isolated from patients with gastric symptoms in Senegal and Cambodia. In Senegal, a significant association was observed between gastric cancer and the cagA gene, two EPIYA-C segments, and the s1 vacA allele in Senegal. Multiple EPIYA-C segments were less frequent than reported in other countries, possibly contributing to the low incidence of gastric cancer. In Cambodia, the frequent introgression of European vacA and cagA alleles into East Asian H. pylori strains was observed. As VacA and CagA have opposing effects, this expansion may disrupt the balancing activities on epithelial cells which might result in severe consequences for individual disease outcome.

Helicobacter pylori est associée à des pathologies gastro-duodénales sévères mais est également un marqueur bactérien de migrations humaines. Nous avons montré que des populations génétiques distinctes de H. pylori ont accompagné au moins quatre migrations en Asie du sud-est et en Océanie : i) une expansion des ancêtres des austronésiens il y a 5000 ans à partir de Taiwan en Océanie, ii) une migration d'Inde en Asie du sud-est depuis 2000 ans,iii) une migration des ancêtres des locuteurs des langues austro-asiatiques au Vietnam et auCambodge il y a 4000 ans, i) une migration des ancêtres des Thaïs du sud de la Chine vers l'actuelle Thaïlande au début du second millénaire. Ces données confirment la résolution plus élevée de la diversité génétique de H. pylori pour retracer les anciennes migrations humaines par comparaison aux marqueurs génétiques humains traditionnels. Nous avons ensuite investigué les facteurs de virulence de souches isolées de patients présentant des symptômes gastriques au Sénégal et au Cambodge. Au Sénégal, une association significative a été observée entre le cancer gastrique et le gène cagA, deux motifs EPIYA-C et l'allèle vacA s1. De multiples segments EPIYA-C étaient observés moins fréquemment que dans les autres régions du monde, contribuant probablement à la faible incidence du cancer gastrique. Au Cambodge, une introgression fréquente d'allèles cagA et vacA européens dans des souches d'Asie de l'est a été observée. CagA et VacA ayant des effets antagonistes, cette expansion pourrait entraîner la rupture de l'équilibre entre les effets biologiques de ces deux protéines et être responsable de conséquences graves sur l'évolution de la maladie.

Submitted by Cássio da Cruz Nogueira (cassionogueirakk@gmail.com) on 2017-10-16T16:24:16Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AssociacaoVariantesRegiao.pdf: 1409940 bytes, checksum: 8e34e8ff0f8dd9a4764a50dee1d96ab2 (MD5) / Approved for entry into archive by Irvana Coutinho (irvana@ufpa.br) on 2017-10-18T14:50:30Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AssociacaoVariantesRegiao.pdf: 1409940 bytes, checksum: 8e34e8ff0f8dd9a4764a50dee1d96ab2 (MD5) / Made available in DSpace on 2017-10-18T14:50:30Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_AssociacaoVariantesRegiao.pdf: 1409940 bytes, checksum: 8e34e8ff0f8dd9a4764a50dee1d96ab2 (MD5) Previous issue date: 2012 / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / A citotoxina CagA do Helicobacter pylori, codificada pelo gene cagA, é associada ao aumento da resposta inflamatória do tecido gástrico e a alteração do controle do crescimento e proliferação celular. A ativação desta citotoxina ocorre pela fosforilação em resíduos específicos de tirosina dentro de uma sequência de aminoácidos denominada motif EPIYA, sendo quatro os tipos de motifs descritos na literatura (EPIYA-A,-B,-C e-D). Contudo, o sítio EPIYA-C constitui o local mais comum de fosforilação de proteínas CagA das cepas bacterianas isoladas nos países ocidentais, podendo ainda ser encontrado em repetições. Assim, este estudo teve como objetivo determinar os tipos de motifs EPIYA de CagA presentes em pacientes com gastrite e adenocarcinoma gástrico e sua associação com estas doenças. Foram coletadas amostras de biópsias gástricas de 384 pacientes infectados por H. pylori, dos quais 194 apresentavam gastrite crônica e 190 adenocarcinoma gástrico. As biópsia gástrica foram utilizadas para análise histológica, extração de DNA bacteriano e análise do gene cagA por PCR. Houve predomínio de adenocarcinoma gástrico no sexo masculino, com média de idade de 58 anos. O gene cagA foi mais prevalente nos pacientes com câncer gástrico, apresentando associação com maior grau de inflamação, atividade neutrofílica e desenvolvimento de metaplasia intestinal (OR = 4,31, IC 95% = 2,71-6,87, p <10-3; OR = 3,57, IC 95% = 2,18 – 5,84, p <10-3; OR = 11,11, IC 95% = 5,48 – 22,30, p <10-3; OR = 3,65, IC 95% = 1,50-8,88, p=0,004, respectivamente). O número de repetições do sítio EPIYA C foi significativamente associada com o aumento do risco de carcinoma gástrico (OR = 2,99, IC 95% = 1,53-5,82, p <10-3) e o maior número de motifs EPIYA C também foi associado com metaplasia intestinal (p = 0,02). Neste estudo a infecção por cepas de H. pylori portadoras do gene cagA com mais de um motif EPIYA-C demonstrou associação com o desenvolvimento de metaplasia intestinal e adenocarcinoma gástrico, entretanto, não apresentou associação com a atividade neutrofílica e grau de inflamação. / Helicobacter pylori CagA cytotoxin, encoded by the cagA gene, has been associated with increased inflammatory response in gastric tissue and the change in control of cell growth and proliferation. Activation of this cytotoxin occurs by phosphorylation in specific tyrosine residues within an amino acid sequence termed motif EPIYA, four types of motifs are described in the literature (EPIYA-A,-B-C and D). However, the site EPIYA-C is the most common site of phosphorylation of CagA protein of the bacterial strains isolated in Western countries, may still be found in repetitions. This study aimed to determine the types of motifs EPIYA of CagA present in patients with gastritis and gastric cancer and its association with these diseases. Were collected samples from gastric biopsies of 384 patients infected with H. pylori, of this 194 presented chronic gastritis and 190 had gastric cancer. The gastric biopsy was used for bacterial DNA extraction and analysis of the cagA gene by PCR. The prevalence of gastric cancer occurs in males, mean age 58 years. The cagA gene was more prevalent in patients with gastric cancer, showing association with a higher degree of inflammation, neutrophil activity and development of intestinal metaplasia (OR = 4,31, IC 95% = 2,71-6,87, p <10-3; OR = 3,57, IC 95% = 2,18 – 5,84, p <10-3; OR = 11,11, IC 95% = 5,48 – 22,30, p <10-3; OR = 3,65, IC 95% = 1,50-8,88, p=0,004, respectively). The number of repetitions EPIYA-C site was significantly associated with increased risk of gastric cancer (OR = 2,99, IC 95% = 1,53-5,82, p <10-3). The higher number of motifs EPIYA-C was also associated with intestinal metaplasia (p = 0,02). In this study the infection by strains of H. pylori carriers cagA gene with more than one motif EPIYA-C shown to be associated with the development of intestinal metaplasia and gastric cancer, but without an association to neutrophil activity and degree of inflammation.