MICA and KIR: Immunogenetic Factors Influencing Left Ventricular Systolic Dysfunction and Digestive Clinical Form of Chronic Chagas Disease.

Tissue damage observed in the clinical forms of chronic symptomatic Chagas disease seems to have a close relationship with the intensity of the inflammatory process. The objective of this study was to investigate whether the MICA (MHC class I-related chain A) and KIR (killer cell immunoglobulin-like receptors) polymorphisms are associated with the cardiac and digestive clinical forms of chronic Chagas disease. Possible influence of these genes polymorphisms on the left ventricular systolic dysfunction (LVSD) in patients with chronic Chagas heart disease was also evaluated. This study enrolled 185 patients with positive serology for Trypanosoma cruzi classified according to the clinical form of the disease: cardiac (n=107) and digestive (n=78). Subsequently, patients with the cardiac form of the disease were sub-classified as with LVSD (n=52) and without LVSD (n=55). A control group was formed of 110 healthy individuals. Genotyping was performed by polymerase chain reaction-sequence specific oligonucleotide probes (PCR-SSOP). Statistical analyzes were carried out using the Chi-square test and odds ratio with 95% confidence interval was also calculated to evaluate the risk association. MICA-129 allele with high affinity for the NKG2D receptor was associated to the LVSD in patients with CCHD. The haplotype MICA*008~HLA-C*06 and the KIR2DS2-/KIR2DL2-/KIR2DL3+/C1+ combination were associated to the digestive clinical form of the disease. Our data showed that the MICA and KIR polymorphisms may exert a role in the LVSD of cardiac patients, and in digestive form of Chagas disease.

Plasma concentrations of CCL3 and CCL4 in the cardiac and digestive clinical forms of chronic Chagas disease.

The aim of this study was to investigate the plasma levels of the CCL3 and CCL4 chemokines in patients with the cardiac and digestive clinical forms of chronic Chagas disease and in cardiac patients with and without left ventricular systolic dysfunction (LVSD). Plasma samples from 75 patients were evaluated by enzyme-linked immunosorbent assay (ELISA) to confirm infection by T. cruzi. Plasma levels of the CCL3 and CCL4 chemokines were measured using Milliplex® MAP assay (Millipore). There were no significant differences in the levels of CCL3 and CCL4 between patients with the digestive and cardiac clinical forms of Chagas disease. Moreover, no significant differences were found between patients without LVSD and those with LVSD. Higher CCL3 and CCL4 plasma levels were found in patients with LVSD compared to those with the digestive form of the disease. The CCL3 and CCL4 chemokines might not be involved in differential susceptibility to the digestive and cardiac clinical forms of chronic Chagas disease, and it seems they do not influence the development of LVSD.

ABO, Secretor and Lewis histo-blood group systems influence the digestive form of Chagas disease.

Chagas disease, caused by Trypanosoma cruzi, can affect the heart, esophagus and colon. The reasons that some patients develop different clinical forms or remain asymptomatic are unclear. It is believed that tissue immunogenetic markers influence the tropism of T. cruzi for different organs. ABO, Secretor and Lewis histo-blood group systems express a variety of tissue carbohydrate antigens that influence the susceptibility or resistance to diseases. This study aimed to examine the association of ABO, secretor and Lewis histo-blood systems with the clinical forms of Chagas disease. We enrolled 339 consecutive adult patients with chronic Chagas disease regardless of gender (cardiomyopathy: n=154; megaesophagus: n=119; megacolon: n=66). The control group was composed by 488 healthy blood donors. IgG anti-T. cruzi antibodies were detected by ELISA. ABO and Lewis phenotypes were defined by standard hemagglutination tests. Secretor (FUT2) and Lewis (FUT3) genotypes, determined by Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), were used to infer the correct histo-blood group antigens expressed in the gastrointestinal tract. The proportions between groups were compared using the χ2 test with Yates correction and Fisher's exact test and the Odds Ratio (OR) and 95% Confidence Interval (95% CI) were calculated. An alpha error of 5% was considered significant with p-values <0.05 being corrected for multiple comparisons (pc). No statistically significant differences were found for the ABO (X2: 2.635; p-value=0.451), Secretor (X2: 0.056; p-value=0.812) or Lewis (X2: 2.092; p-value=0.351) histo-blood group phenotypes between patients and controls. However, B plus AB Secretor phenotypes were prevalent in pooled data from megaesophagus and megacolon patients (OR: 5.381; 95% CI: 1.230-23.529; p-value=0.011; pc=0.022) in comparison to A plus O Secretor phenotypes. The tissue antigen variability resulting from the combined action of ABO and Secretor histo-blood systems is associated with the digestive forms of Chagas disease.

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.

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, alone 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.

[Cross-cultural adaptation and validation for Portuguese (Brazil) of health related quality of life instruments specific for gastroesophageal reflux disease]. / Tradução e validação para a língua portuguesa (Brasil) de instrumentos específicos para avaliação de qualidade de vida na doença do refluxo gastroesofágico.

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.

Tradução e validação para a língua portuguesa (Brasil) de instrumentos específicos para avaliação de qualidade de vida na doença do refluxo gastroesofágico / Cross-cultural adaptation and validation for Portuguese (Brazil) of health related quality of life instruments specific for gastroesophageal reflux disease

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, 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 sintomßtico 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...

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...

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.

Polymorphisms of DNA repair genes XRCC1 and XRCC3, interaction with environmental exposure and risk of chronic gastritis and gastric 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 relationship 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.

Aneuploidies, deletion, and overexpression of TP53 gene in intestinal metaplasia of patients without gastric cancer.

Gastric carcinogenesis is attributable to interacting environmental and genetic factors, through a sequence of events including intestinal metaplasia. Using a fluorescence in situ hybridization technique, we investigated the occurrence of aneuploidies of chromosomes 3, 7, 8, 9, and 17, TP53 gene deletion, and expression of p53 in 21 intestinal metaplasia (IM) samples from cancer-free patients and in 20 gastric adenocarcinoma samples. Aneuploidies were found in 71% (15/21) of the IM samples. Trisomy of chromosomes 7 and 9 occurred mainly in complete-type IM; in the incomplete type, trisomy of chromosomes 7 and 8 were more commonly found. The TP53 gene deletion was observed in 60% (3/5) of the IM cases, and immunohistochemistry revealed p53 overexpression in 12% (2/17) of the analyzed IM cases. All gastric adenocarcinoma cases presented higher frequencies of trisomy or tetrasomy of chromosomes 3, 7, 8, 9, and 17. The TP53 deletion was found in all three of the gastric adenocarcinoma analyzed for it, and immunohistochemistry detected overexpression of protein p53 in 80% (12/15) of the analyzed cases. Our study revealed for the first time the presence of aneuploidies of chromosomes 7, 8, 9, and 17 and of TP53 gene deletion and overexpression in IM samples from cancer-free patients. These results suggest that IM and gastric adenocarcinoma may share the same genetic alterations.

Cytogenetic alterations in chagasic achalasia compared to esophageal carcinoma.

Patients with chagasic achalasia (megaesophagus) are liable to have an additional 1.7-20% possibility of developing esophageal squamous cell carcinoma (ESCC). We applied a fluorescence in situ hybridization technique in 20 such patients and found aneuploidies of chromosomes 7, 11, and 17 in 60% (12 of 20 specimens) and deletion of the TP53 gene in 54.5% (6 of 11 specimens; it was only possible to obtain data by FISH technique from 11 of the 20 achalasia patients). The main aneuploidies detected were chromosome 7 monosomy or trisomy (35%) in mid-third megaesophagus cases, and chromosome 17 monosomy or trisomy (25%) in distal-third cases. TP53 gene deletion was more frequent in mid-third (62.5%) than in distal-third megaesophagus cases (40%). In chagasic megaesophagus, no amplification of the cyclin D1 gene (CCND1) was observed. Comparing chagasic megaesophagus to ESCC, we found a higher frequency of aneuploidies in all 10 tumors. The main alterations were trisomy or tetrasomy of chromosomes 17 (90%), 11 (70%), and 7 (70%). Amplification of CCND1 was evidenced as a cluster in 70% of the tumors (22-99% of nuclei), while TP53 gene deletion occurred in 100%. To our knowledge, this is the first cytogenetic analysis of chagasic megaesophagus to show that aneuploidies of chromosomes 7, 11, and 17, and TP53 gene deletion might be related to increased risk for malignancy.

GSTT1, GSTM1 and CYP2E1 genetic polymorphisms in gastric cancer and chronic gastritis in a Brazilian population.

AIM: To test the hypothesis that, in the Southeastern Brazilian population, the GSTT1, GSTM1 and CYP2E1 polymorphisms and putative risk factors are associated with an increased risk for gastric cancer. METHODS: We conducted a study on 100 cases of gastric cancer (GC), 100 cases of chronic gastritis (CG), and 150 controls (C). Deletion of the GSTT1 and GSTM1 genes was assessed by multiplex PCR. CYP2E1/PstI genotyping was performed using a PCR-RFLP assay. RESULTS: No relationship between GSTT1/GSTM1 deletion and the c1/c2 genotype of CYP2E1 was observed among the three groups. However, a significant difference between CG and C was observed, due to a greater number of GSTT1/GSTM1 positive genotypes in the CG group. The GSTT1 null genotype occurred more frequently in Negroid subjects, and the GSTM1 null genotype in Caucasians, while the GSTM1 positive genotype was observed mainly in individuals with chronic gastritis infected with H pylori. CONCLUSION: Our findings indicate that there is no obvious relationship between the GSTT1, GSTM1 and CYP2E1 polymorphisms and gastric cancer.