kDNA gene signatures of Trypanosoma cruzi in blood and oesophageal mucosa from chronic chagasic patients

Trypanosoma cruzi presents a high degree of intraspecific variability, with possible implications for the pathogenesis of Chagas disease. The aim of this study was to evaluate T cruzi kDNA minicircle gene signatures using the low-stringency single-specific-primer PCR technique in both peripheral blood and oesophageal. mucosa from chronic chagasic patients, with or without megaesophagus, atone or in combination with cardiopathy and megacolon. It was not possible to identify a uniform pattern of shared bands between blood and oesophageal mucosa samples from individuals with the same clinical. form or mixed forms, suggesting multiple T. cruzi infections with differential tissue tropism. Thus, the results indicate that there is an intense intraspecific variability in the hypervariable regions of T cruzi kDNA, which has so far made it impossible to correlate the genetic profile of this structure with the clinical manifestations of Chagas disease. (C) 2008 Royal. Society of Tropical Medicine and Hygiene. Published by Elsevier Ltd. AIL rights reserved

Polymorphisms of DNA repair genes XRCC1 and XRCC3, interaction with environmental exposure and risk of chronic gastritis and grastic cancer

Aim: To evaluate the association between polymorphisms XRCC1 Arg194Trp and Arg399Gln and XRCC3 Thr241Met and the risk for chronic gastritis and gastric cancer, in a Southeastern Brazilian population. Methods: Genotyping by PCR-RFLP was carried out on 202 patients with chronic gastritis (CG) and 160 patients with gastric cancer (GC), matched to 202 (C1) and 150 (C2) controls, respectively. Results: No differences were observed among the studied groups with regard to the genotype distribution of XRCC1 codons 194 and 399 and of XRCC3 codon 241. However, the combined analyses of the three variant alleles (194Trp, 399Gln and 241Met) showed an increased risk for chronic gastritis when compared to the GC group. Moreover, an interaction between the polymorphic alleles and demographic and environmental factors was observed in the CG and GC groups. XRCC1 194Trp was associated with smoking in the CG group, while the variant alleles XRCC1 399Gln and XRCC3 241Met were related with gender, smoking, drinking and H pylori infection in the CG and GC groups. Conclusion: Our results showed no evidence of a rela-tionship between the polymorphisms XRCC1 Arg194Trp and Arg399Gln and XRCC3 Thr241Met and the risk of chronic gastritis and gastric cancer in the Brazilian population, but the combined effect of these variants may interact to increase the risk for chronic gastritis, considered a premalignant lesion. Our data also indicate a gene-environment interaction in the susceptibility to chronic gastritis and gastric cancer. © 2005 The WJG Press and Elsevier Inc. All rights reserved

Genetic alterations in benign lesions: Chronic gastritis and gastric ulcer

Aim: To investigate the occurrence of chromosome 3, 7, 8, 9, and 17 aneuploidies, TP53 gene deletion and p53 protein expression in chronic gastritis, atrophic gastritis and gastric ulcer, and their association with H pylori infection. Methods: Gastric biopsies from normal mucosa (NM, n = 10), chronic gastritis (CG, n = 38), atrophic gastritis (CAG, n = 13) and gastric ulcer (GU, n = 21) were studied using fluorescence in situ hybridization (FISH) and immunohistochemical assay. A modified Giemsa staining technique and PCR were used to detect H pylori. An association of the gastric pathologies and aneuploidies with H pylori infection was assessed. Results: Aneuploidies were increasingly found from CG (21%) to CAG (31%) and to GU (62%), involving mainly monosomy and trisomy 7, trisomies 7 and 8, and trisomies 7, 8 and 17, respectively. A significant association was found between H pylori infection and aneuploidies in CAG (P = 0.0143) and GU (P = 0.0498). No TP53 deletion was found in these gastric lesions, but p53-positive immunoreactivity was detected in 45% (5/11) and 12% (2/17) of CG and GU cases, respectively. However, there was no significant association between p53 expression and H pylori infection. Conclusion: The occurrence of aneuploidies in benign lesions evidences chromosomal instability in early stages of gastric carcinogenesis associated with H pylori infection, which may confer proliferative advantage. The increase of p53 protein expression in CG and GU may be due to overproduction of the wild-type protein related to an inflammatory response in mucosa. © 2006 The WJG Press. All rights reserved

Cross-cultural adaptation and validation for Portuguese (Brazil) of health related quality of life instruments specific for gastroesophageal reflux disease

BACKGROUND: Gastroesophageal reflux disease has been shown patients to alter quality of life and working productivity. Most of the instruments available for this type of assessment come from English or French speaking countries. To use these instruments in Brazil requires a judicious process of translation and validation. AIM: Translating to Portuguese the questionnaires GSAS (Gastroesophageal Reflux Disease Symptom Assessment Scale), GERD-HRQL (Gastroesophageal Reflux Disease - Health Related Quality of Life) and HBQOL (Heartburn Specific Quality of Life Instrument) specific for quality of life assessment in gastroesophageal reflux disease. Testing the psychometric properties of reliability and validity of the referred disease specific instruments. METHODS: One hundred and thirty two gastroesophageal reflux disease patients (mean age 54.9 years and ± SD 13.9) from the Digestive Disease Motility Outpatient Clinic, Federal University of São Paulo, SP, Brazil and the Department of Surgical Gastroenterology São José do Rio Preto School of Medicine, São José do Rio Preto, SP, Brazil, accepted to participate and signed the informed consent form. Forty of these patients took part in the pre-test phase (28 females and 12 males, mean age 55.3 years ± SD 14.7) and the remaining 92 part in the validation phase (64 females and 28 males, mean age 54.7 years and ± SD 13.7). The translation and cultural adaptation processes were carried out accordingly us to the method of Guillemin et al (1993). The validation processes of the disease specific translated questionnaires (GSAS, GERD-HRQL and HBQOL) was performed in relation to a generic (SF-36) and a symptomatic (SQGERD) instrument. RESULTS: Nine words of the GSAS, four of the GERD-HRQL and six of the HBQOL were replaced during the cultural adaptation phase. The GSAS questionnaire was discontinued after this phase because of scoring problems. Therefore reliability and validity were tested only for the two remaining questionnaires. These questionnaires proved to be reproducible for both inter and intra-observer relationships (0.980 and 0.968 values for the GERD-HRQL and varying values of 0.868 to 0.972 for the HBQOL). The HBQOL questionnaire demonstrated high internal consistency (>0.70) for three of the four dimensions tested (physical aspect, pain, sleep). Good correlations levels with the SF-36 and SQGERD questionnaires were demonstrated during the validation phase. CONCLUSIONS: The cross cultural adaptation of the Portuguese (Brazil) versions of the GERD-HRQL and HBQOL instruments proved to be reliable and valid options with low burden level for assessment of quality of life in gastroesophageal reflux disease our country. The HBQOL is the only multidimensional questionnaire for quality of life assessment in gastroesophageal reflux disease currently available in Brazil. The Portuguese (Brazil) version of the GSAS instrument proved inadequate for quality of life assessment in our country.INTRODUÇÃO: Estudos têm demonstrado ser a doença do refluxo gastroesofágico capaz de alterar a qualidade de vida e a produtividade no trabalho dos doentes por ela acometidos. Instrumentos para esse tipo de avaliação são provenientes, em sua maioria, de países de língua inglesa e/ou francesa. A utilização desses instrumentos em nosso meio demanda criterioso processo de tradução e validação. OBJETIVOS: Traduzir para língua portuguesa os questionários GERD-HRQL (Gastroesophageal Reflux Disease - Health Related Quality of Life), HBQOL (Heartburn Specific Quality of Life Instrument) e GSAS (Gastroesophageal Reflux Disease Symptom Assessment Scale) específicos para avaliação de qualidade de vida na doença do refluxo gastroesofágico. Testar suas propriedades psicométricas de confiabilidade e validade. MÉTODOS: Cento e trinta e dois pacientes com doença do refluxo gastroesofágico (idade média 54,9 anos, ± DP 13,9) atendidos no ambulatório de motilidade digestiva da Universidade Federal de São Paulo (UNIFESP), SP, e de gastrocirurgia da Faculdade de Medicina de São José do Rio Preto, SP, aceitaram participar do presente estudo, fornecendo termo de consentimento pós-esclarecimento. Destes, 40 pacientes participaram da fase de pré-teste (28 do sexo feminino e 12 do sexo masculino, com idade média de 55,3 anos, ± DP 14,7) e 92 da fase de validação (64 do sexo feminino e 28 sexo masculino, com idade média 54,7 anos e ± DP 13,7). A tradução e adaptação cultural foi realizada de acordo com o método de GUILLEMIN et al., sendo a validação dos questionários traduzidos (GERD-HRQL, HBQOL e GSAS) realizada em relação aos instrumentos genérico SF-36 e sintomático ESDRGE (SQGERD). RESULTADOS: A adaptação cultural implicou na troca de quatro palavras no GERD-HRQL, seis no HBQOL e nove no GSAS. Posteriormente a esta fase, o questionário GSAS foi abandonado por problemas no cálculo do escore, sendo as propriedades de medidas testadas nos dois questionários remanescentes, esses se mostraram reprodutíveis para uso inter e intra-observador com valores de 0,980 e 0,968, respectivamente, para o GERD-HRQL, e valores que variaram de 0,868 a 0,972, respectivamente, para o HBQOL. O questionário HBQOL demonstrou alta consistência interna (>0,70) para três das quatro dimensões avaliadas (aspecto físico, dor, sono). Os resultados encontrados na fase de validação apresentaram bons níveis de correlação com os questionários SF-36 e ESDRGE (SQGERD). CONCLUSÕES: As versões para a língua portuguesa (Brasil) dos instrumentos GERD-HRQL e HBQOL, adaptadas ao padrão cultural brasileiro, configuram-se em opções válidas, confiáveis, com baixo nível de desgaste do paciente e de fácil aplicação para avaliação de qualidade de vida na DRGE em nosso meio. O instrumento HBQOL é a única opção de avaliação multidimensional de qualidade de vida atualmente disponível para uso no Brasil. A versão em português do instrumento GSAS mostrou-se inadequada para avaliação de qualidade de vida na DRGE em nosso meio.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Federal University of São PauloSão José do Rio Preto School of MedicineUniversidade Federal de São Paulo (UNIFESP) Department of MedicineFAMERP Department of SurgeryUNIFESPUNIFESP, Department of MedicineUNIFESPSciEL

Genetic Susceptibility to Cardiac and Digestive Clinical Forms of Chronic Chagas Disease: Involvement of the CCR5 59029 A/G Polymorphism.

The clinical manifestations of chronic Chagas disease include the cardiac form of the disease and the digestive form. Not all the factors that act in the variable clinical course of this disease are known. This study investigated whether the CCR5Δ32 (rs333) and CCR5 59029 A/G (promoter region--rs1799987) polymorphisms of the CCR5 gene are associated with different clinical forms of chronic Chagas disease and with the severity of left ventricular systolic dysfunction in patients with chronic Chagas heart disease (CCHD). The antibodies anti-T. cruzi were identified by ELISA. PCR and PCR-RFLP were used to identify the CCR5Δ32 and CCR5 59029 A/G polymorphisms. The chi-square test was used to compare variables between groups. There was a higher frequency of the AA genotype in patients with CCHD compared with patients with the digestive form of the disease and the control group. The results also showed a high frequency of the AG genotype in patients with the digestive form of the disease compared to the other groups. The results of this study show that the CCR5Δ32 polymorphism does not seem to influence the different clinical manifestations of Chagas disease but there is involvement of the CCR5 59029 A/G polymorphism in susceptibility to the different forms of chronic Chagas disease. Besides, these polymorphisms do not influence left ventricular systolic dysfunction in patients with CCHD