Marialva Tereza Ferreira de Araújo

Prevalência de genótipos e de mutantes pré-core A-1896 do vírus da hepatite B e suas implicações na hepatite crônica, em uma população da Amazônia oriental

A infeccao pelo virus da hepatite B apresenta amplo espectro de manifestacoes clinicas. Objetivando conhecer os genotipos do HBV mais prevalentes e determinar a ocorrencia da mutacao pre-core A-1896, em uma populacao da Amazonia oriental, correlacionando com o diagnostico clinico, foram selecionados 51 pacientes portadores cronicos de HBsAg e HBV-DNA positivos e divididos em tres grupos: grupo A (n=14, pacientes assintomaticos); grupo B (n=20, sintomaticos HBeAg positivos) e grupo C (n=17, sintomaticos HBeAg negativos), sendo usado o sequenciador automatico ABI modelo 377 para identificacao de genotipos e mutantes pre-core. Os resultados evidenciaram o genotipo A como o mais prevalente, 81,8%, 89,5% e 93,7%, nos grupos A, B e C, respectivamente. A mutacao pre-core A-1896 foi encontrada em 11,5% (3/26), sendo todos assintomaticos. Concluiu-se que na populacao estudada o genotipo A foi o mais prevalente e houve baixa ocorrencia do mutante pre-core A-1896, ambos nao se constituindo fatores agravantes da doenca hepatica.

Infecções pelos vírus das hepatites B e C e o carcinoma hepatocelular na Amazônia oriental

Com o objetivo de contribuir para um melhor conhecimento do envolvimento das infeccoes pelos virus das hepatites B e C, na etioepidemiologia do CHC na Amazonia Oriental, estudou-se 36 pacientes em Belem-PA. Foram avaliados marcadores sorologicos e a pesquisa do HBV-DNA e HCV-RNA pela reacao em cadeia da polimerase. Observou-se etilismo em 33,3% e cirrose em 83,3%. Marcadores sorologicos das infeccoes pelo HBV e HCV foram encontrados respectivamente em 88,9% e 8,3%. O HBsAg foi encontrado em 58,3%; anti-HBc em 86%; anti-HBe em 85,7; HBeAg em 9,5%; anti-HBc IgM em 57,1%. O HBV-DNA foi detectado em 37,7% e em 65% dos HBsAg positivos; o HCV-RNA em 8,5% e em 100% dos anti-HCV positivos. AFP esteve alterada em 88,9% e acima de 400ng/ml em 75% dos casos. Conclui-se que a infeccao pelo HBV parece ter importância na etiologia do CHC e ressalta-se a importância de implementar programas de vacinacao e deteccao precoce do tumor.

Lesão intestinal após isquemia-reperfusão: estudo comparativo usando sal tetrazólico (MTT) e histologia

Varios metodos sao utilizados para avaliar e estimar as lesoes intestinais de isquemia e reperfusao (IR). Assim, o objetivo do presente trabalho e realizar o estudo comparativo dos aspectos colorimetrico e histologico da lesao intestinal apos IR. Para tal, foram utilizados 30 ratos Wistar, machos, pesando entre 310 a 410g, distribuidos em 3 grupos: Grupo Controle (GC), Grupo Isquemia e Reperfusao-1 (GIR-1) e Grupo Isquemia e Reperfusao-3 (GIR-3), com 10 animais cada. Nos grupos GIR-1 e GIR-3 foi realizada isquemia intestinal, por meio de falsa ligadura da arteria mesenterica anterior, durante 30 minutos e apos esta a perfusao sanguinea foi restaurada. Estes animais foram submetidos a eutanasia apos 1 e 3 dias de reperfusao, respectivamente, sendo colhido material para realizacao dos estudos colorimetrico, usando o Methyl Thiazolyl Blue (MTT) e histologico pela hematoxilina e eosina. Os resultados obtidos demonstraram uma menor proporcao de celulas viaveis e um maior grau de lesao da tunica mucosa nos animais do grupo GIR-3 em relacao ao controle (p<0,05). Desta forma os autores concluem que o estudo colorimetrico, usando o MTT, mostrou-se tao eficaz e confiavel quanto o estudo histologico na avaliacao das repercussoes intestinais produzidas pela IR deste orgao.

Association of killer cell immunoglobulin-like receptor polymorphisms with chronic hepatitis C and responses to therapy in Brazil

Soroprevalence for Hepatitis C virus is reported as 2.12% in Northern Brazil, with about 50% of the patients exhibiting a sustained virological response (SVR). Aiming to associate polymorphisms in Killer Cell Immunoglobulin-like Receptors (KIR) with chronic hepatitis C and therapy responses we investigated 125 chronic patients and 345 controls. Additionally, 48 ancestry markers were genotyped to control for population stratification. The frequency of the KIR2DL2 and KIR2DL2+HLA-CAsp80 gene and ligand was higher in chronic infected patients than in controls (p < 0.0009, OR = 3.4; p = 0.001, OR = 3.45). In fact, KIR2DL3 is a weaker inhibitor of NK activity than KIR2DL2, which could explain the association of KIR2DL2 with chronic infection. Moreover, KIR2DS2 and KIR2DS2+HLA-CAsp80 (p < 0.0001, OR = 2.51; p = 0.0084, OR = 2.62) and KIR2DS3 (p < 0.0001; OR = 2.57) were associated with chronic infection, independently from KIR2DL2. No differences in ancestry composition were observed between control and patients, even with respect to therapy response groups. The allelic profile KIR2DL2/KIR2DS2/KIR2DS3 was associated with the chronic hepatitis C (p < 0.0001; OR = 3). Furthermore, the patients also showed a higher mean number of activating genes and a lower frequency of the homozygous AA profile, which is likely secondary to the association with non-AA and/or activating genes. In addition, the KIR2DS5 allele was associated with SVR (p = 0.0261; OR = 0.184).The ancestry analysis of samples ruled out any effects of population substructuring and did not evidence interethnic differences in therapy response, as suggested in previous studies.

Expressão dos antígenos ABH e Lewis na gastrite crônica e alterações pré- neoplásica da mucosa gástrica

RESUMO – Racional - A aderencia do Helicobacter pylori a mucosa gastrica humana e pre-requisito para sua colonizacao e o desenvolvimento da gastrite cronica. Os antigenos de grupos sanguineos, presentes no muco gastrico, sao descritos como provaveis receptores da bacteria neste epitelio. A expressao alterada destes antigenos esta associada ao desenvolvimento do câncer gastrico. Objetivos - Verificar a ocorrencia do Helicobacter pylori e a distribuicao da expressao dos antigenos ABH e Lewis correlacionada com as alteracoes histopatologicas de pacientes com gastrite cronica. Pacientes e Metodos – Analisaram-se 63 amostras de sangue, saliva e biopsias gastricas de pacientes com gastrite cronica atraves das tecnicas dot-blot-ELISA, imunoperoxidase indireta e coloracoes do Gram modificado e hematoxilina-eosina. Resultados - Nao foram encontradas associacoes significativas entre a presenca da bacteria e os fenotipos de grupos sanguineos ABH, Lewis e Secretor. Na maioria dos pacientes, a expressao dos antigenos ABH e Lewis, estava restrita principalmente ao epitelio foveolar da mucosa gastrica, concordando com a expressao ao nivel salivar. A expressao inapropriada desses antigenos ocorria sempre na infeccao pelo Helicobacter pylori e/ou alteracoes pre-neoplasicas da mucosa gastrica. Em areas com metaplasia intestinal foi observada a reducao da reatividade para os antigenos H e Leb, e principalmente o aumento de Lea. Conclusao - Alteracoes no padrao de glicosilacao destes antigenos refletem diferentes estagios de diferenciacao celular e sao marcadores potenciais na avaliacao diagnostica e prognostica das patologias gastricas. DESCRITORES – Helicobacter pylori. Infeccoes por helicobacter. Gastrite. Grupos sanguineos.

SNP rs8099917 in Gene IL28B Might Be Associated with Risk of Chronic Infection by HCV but Not with Response to Treatment

Aim. The aim of this study was to characterize the genetic profile of patients with chronic hepatitis C virus (HCV) infection relative to polymorphisms rs12979860 and rs8099917 in gene IL28B and the association of those polymorphisms with the response to treatment with pegylated interferon and ribavirin, performed at a reference center in Brazilian Amazonia. Methods. A total of 75 individuals with chronic hepatitis C and 98 healthy individuals from both genders over 18 years old were assessed. DNA samples were collected from leukocytes and subjected to real-time polymerase chain reaction to genotype polymorphisms rs12979860 and rs8099917. Results. Analysis of the allelic and genotypic frequencies of the investigated polymorphisms showed that both groups were in Hardy-Weinberg equilibrium; polymorphism rs12979860 exhibited no significant difference between the groups. For polymorphism rs8099917, allele T was significantly less frequent (P = 0.0195) among the patients (63.3%) than the controls (75.5%), and the patients were 1.7 times as likely to exhibit allele G. No difference in response to treatment was associated with SNP patterns. Conclusion. The results suggest a possible association of SNP rs8099917 with higher odds of chronic HCV infection but do not indicate a putative influence of the investigated SNPs on the sustained virologic response.

Intrahepatic mRNA Expression of FAS, FASL, and FOXP3 Genes Is Associated with the Pathophysiology of Chronic HCV Infection

This study aimed to evaluate the relative mRNA expression of Fas receptor (FAS), Fas ligand (FASL), and forkhead box protein 3 (FOXP3) in liver biopsy specimens obtained from patients with viral and non-viral chronic hepatitis and correlate their expression with the fibrosis stage. A total of 51 liver biopsy specimens obtained from HBV (n = 6), HCV (n = 28), and non-viral hepatic disease (NVHD) (n = 9) patients and from individuals with normal liver histology (n = 8) (control—CT) were analyzed. Quantifications of the target genes were assessed using qPCR, and liver biopsies according to the METAVIR classification. The mRNA expression levels of FAS and FASL were lower in the CT group compared to the groups of patients. The increase in the mRNA expression of FAS and FASL was correlated with higher levels of inflammation and disease progression, followed by a decline in tissues with cirrhosis, and it was also associated with increased levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST). Higher mRNA expression of FOXP3 was observed in the HCV and NVHD groups, with the peak observed among patients with cirrhosis. The increased FOXP3 mRNA expression was positively correlated with increased FAS and FASL mRNA expression and the AST and ALT levels in all patients. Conclusions: These results suggest that regardless of the cause, the course of chronic liver disease may be modulated by the analyzed genes and correlated with an increase in regulatory T cells during the liver damage followed by hepatocyte destruction by Fas/FasL system and subsequent non specific lymphocytic infiltrate accumulation.

Absence of Correlation between IL-28B Gene Polymorphisms and the Clinical Presentation of Chronic Hepatitis B in an Amazon Brazilian Population

Objective. The present study investigated the prevalence of the IL-28B polymorphisms rs12979860 and rs8099917 in chronic hepatitis B patients from a case study in Eastern Amazonia. Methods. In total, 65 chronically infected HBV patients and 97 healthy subjects who were anti-HBc and anti-HBs positive (control group) were evaluated between May 2011 and December 2012. The groups of patients were designated as inactive carriers, chronic hepatitis without cirrhosis, and chronic hepatitis with cirrhosis based on clinical, pathological, biochemical, hematological, and virological variables. The patients were genotyped using quantitative real-time PCR. Results. The frequencies of the rs12979860 polymorphism were similar between the infected group (32.3% CC, 41.5% CT, and 26.2 TT) and the control population (35% CC, 47.4% CT, and 17.6% TT), and the frequencies of the rs8099917 polymorphism (7.7% GG, 35.4% GT, and 56.9% TT versus 7.2% GG, 35.1% GT, and 57.7% TT) were also similar in both groups. The associations between the rs12979860 and rs8099917 polymorphisms and the clinical manifestations were not statistically significant. Conclusion. In conclusion, these polymorphisms had a similar distribution between infected and control groups, indicating that they were not associated with susceptibility and the clinical evolution of hepatitis B in the examined population.

NGF and P75NTR Gene Expression Is Associated with the Hepatic Fibrosis Stage Due to Viral and Non-Viral Causes

This study evaluated the relative mRNA expression levels of nerve growth factor (NGF) and the p75 neurothrophin receptor (p75NTR) in different histological stages of human liver disease. Fifty-one liver biopsy specimens obtained from patients with hepatitis B virus (n = 6), hepatitis C virus (n = 28), and non-viral hepatitis – (n = 9) and standard histological liver (n = 8) as controls (CT) were subjected to qPCR and histopathological exams. Our data revealed a significant difference in the NGF expression levels between the three patient groups and the Control group. p75NTR expression levels in the HCV and NVH groups were higher than those observed in the HBV and Control groups. In cases of liver cirrhosis, higher p75NTR mRNA expression was observed, whereas NGF was expressed at higher levels in patients with hepatic fibrosis. NGF expression was lower in the F1 liver fibrosis stage, and p75NTR receptor expression continuously and proportionately increased compared to the increase in the degree of fibrosis and was significantly higher in livers in fibrosis stages 3 and 4. The hepatic levels of NGF and p75NTR were decreased and increased, respectively, relative to the stage of inflammatory activity. A positive correlation between p75NTR and NGF gene expression was observed in livers with mild to moderate fibrosis, though not in cases of severe fibrosis and cirrhosis. Conclusion Our results demonstrate that the course of chronic liver disease can be regulated by NGF and p75NTR, which function by decreasing or inhibiting hepatocyte regeneration and proliferation.

Programa de hepatopatias do Hospital da Fundação Santa Casa de Misericórdia do Pará: infra-estrutura e epidemiologia clínica em 10 anos de atendimento

O Programa de Hepatopatias do Hospital da Fundacao Santa Casa de Misericordia do Para surgiu pela necessidade de prestar assistencia a hepatopatas na regiao amazonica priorizando assistencia qualificada, identificacao das etiologias, seguimento clinico, e tratamento direcionado. Este trabalho visa descrever dados relativos a epidemiologia clinica, fatores etiologicos e analise histopatologica. Dos 1469 pacientes avaliados, atraves de exames clinicos, laboratoriais, endoscopicos e de imagem e/ou histopatologico, foram considerados hepatopatas cronicos 935 (63,6%). Nesta casuistica, a media de idade foi 50 anos, 666 (71,2%) do sexo masculino e maior procedencia de Belem. Os agentes etiologicos mais prevalentes foram alcoolismo (53,7%) e hepatites virais (39,1%). Biopsia hepatica realizada em 403/935 (43,1%), demonstrou hepatite cronica (34%) e cirrose (34%) na maioria das amostras. Conclui-se, portanto, que a doenca hepatica cronica na regiao e mais prevalente no sexo masculino, sendo o alcoolismo a principal etiologia e mais da metade dos casos se encontravam em fase avancada no momento do diagnostico.