Recurrence of hepatitis C virus after treatment with pegylated interferon and direct acting antivirals in Punjab Pakistan / Recorrência do vírus da hepatite C após tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão

Although increased response rates concomitant in hepatitis C virus but relapse after treatment is threatened. Therefore, it is terrible requirement to evaluate the response of Pegylated interferon and direct acting antivirals in Punjab Pakistan. The study was conducted to find the rate of recurrence of HCV infection after treatment with Pegylated Interferon and Direct Acting Antivirals in Punjab Pakistan. This study was conducted at Department of Pathology, Nawaz Sharif Medical College Gujrat, while treatment effects monitored in different Government and Private Hospitals of Punjab, Pakistan. Total 973 patients who administered the recommended dose and divided in two groups (i) Interferon based therapy (ii) direct acting antivirals (DAAs).Other parameters like ALT and viral load studied. The rate of recurrence was higher in female infected with genotype 2b and in male with mixed genotype 3a/2b after six month of antiviral therapy. Genotype 3a showed significant response to therapy after three month. 32 among 374 (8.5%) were positive after 24 weeks of treatment with interferon, 29 (7.7%) patients have same genotype while 3 patients were re-infected with different HCV strains. With DAAs, only 27 (4.8%) patients were positive among 558 after 2 weeks and one patient re-infected with different genotype. Early and sustained virological response noted in DAAs. ALT and viral load decreased faster with DAAs that not achieved after 4 weeks with pegylated interferon. Sustained virological response appears in DAAs and recurrence rate is high in interferon therapy compared to DAAs. Therefore, reinfection has implications for correct treatment efficiency and to select strategies for retreatment cases.

Embora aumentem as taxas de resposta concomitantes no vírus da hepatite C (HCV), há risco de recidiva após o tratamento. Portanto, é um requisito terrível avaliar a resposta do interferon peguilado e antivirais de ação direta em Punjab, Paquistão. O estudo foi conduzido para encontrar a taxa de recorrência da infecção por HCV após o tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão. Este estudo foi conduzido no Departamento de Patologia Nawaz Sharif Medical College Gujrat, enquanto os efeitos do tratamento foram monitorados em diferentes hospitais públicos e privados de Punjab, Paquistão. Total de 973 pacientes que administraram a dose recomendada foram divididos em dois grupos: (i) Terapia baseada em interferon, (ii) antivirais de ação direta (DAAs). Outros parâmetros como ALT e carga viral foram estudados. A taxa de recorrência foi maior em mulheres infectadas com o genótipo 2b e em homens com genótipo misto 3a / 2b após seis meses de terapia antiviral. O genótipo 3a mostrou resposta significativa à terapia após três meses. 32 entre 374 (8,5%) foram positivos após 24 semanas de tratamento com interferon, 29 (7,7%) pacientes têm o mesmo genótipo, enquanto 3 pacientes foram reinfectados com diferentes cepas de HCV. Com DAAs, apenas 27 (4,8%) pacientes foram positivos entre 558 após duas semanas e um paciente reinfectado com genótipo diferente. Resposta virológica precoce e sustentada observada em DAAs. ALT e carga viral diminuíram mais rapidamente com DAAs, que não alcançou após 4 semanas com interferon peguilado. A resposta virológica sustentada aparece em DAAs, e a taxa de recorrência é alta na terapia com interferon em comparação com DAAs. Portanto, a reinfecção tem implicações para a eficiência do tratamento correto e para selecionar estratégias para casos de retratamento.

Recurrence of hepatitis C virus after treatment with pegylated interferon and direct acting antivirals in Punjab Pakistan

Abstract Although increased response rates concomitant in hepatitis C virus but relapse after treatment is threatened. Therefore, it is terrible requirement to evaluate the response of Pegylated interferon and direct acting antivirals in Punjab Pakistan. The study was conducted to find the rate of recurrence of HCV infection after treatment with Pegylated Interferon and Direct Acting Antivirals in Punjab Pakistan. This study was conducted at Department of Pathology, Nawaz Sharif Medical College Gujrat, while treatment effects monitored in different Government and Private Hospitals of Punjab, Pakistan. Total 973 patients who administered the recommended dose and divided in two groups (i) Interferon based therapy (ii) direct acting antivirals (DAAs).Other parameters like ALT and viral load studied. The rate of recurrence was higher in female infected with genotype 2b and in male with mixed genotype 3a/2b after six month of antiviral therapy. Genotype 3a showed significant response to therapy after three month. 32 among 374 (8.5%) were positive after 24 weeks of treatment with interferon, 29 (7.7%) patients have same genotype while 3 patients were re-infected with different HCV strains. With DAAs, only 27 (4.8%) patients were positive among 558 after 2 weeks and one patient re-infected with different genotype. Early and sustained virological response noted in DAAs. ALT and viral load decreased faster with DAAs that not achieved after 4 weeks with pegylated interferon. Sustained virological response appears in DAAs and recurrence rate is high in interferon therapy compared to DAAs. Therefore, reinfection has implications for correct treatment efficiency and to select strategies for retreatment cases.

RESUMO Embora aumentem as taxas de resposta concomitantes no vírus da hepatite C (HCV), há risco de recidiva após o tratamento. Portanto, é um requisito terrível avaliar a resposta do interferon peguilado e antivirais de ação direta em Punjab, Paquistão. O estudo foi conduzido para encontrar a taxa de recorrência da infecção por HCV após o tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão. Este estudo foi conduzido no Departamento de Patologia Nawaz Sharif Medical College Gujrat, enquanto os efeitos do tratamento foram monitorados em diferentes hospitais públicos e privados de Punjab, Paquistão. Total de 973 pacientes que administraram a dose recomendada foram divididos em dois grupos: (i) Terapia baseada em interferon, (ii) antivirais de ação direta (DAAs). Outros parâmetros como ALT e carga viral foram estudados. A taxa de recorrência foi maior em mulheres infectadas com o genótipo 2b e em homens com genótipo misto 3a / 2b após seis meses de terapia antiviral. O genótipo 3a mostrou resposta significativa à terapia após três meses. 32 entre 374 (8,5%) foram positivos após 24 semanas de tratamento com interferon, 29 (7,7%) pacientes têm o mesmo genótipo, enquanto 3 pacientes foram reinfectados com diferentes cepas de HCV. Com DAAs, apenas 27 (4,8%) pacientes foram positivos entre 558 após duas semanas e um paciente reinfectado com genótipo diferente. Resposta virológica precoce e sustentada observada em DAAs. ALT e carga viral diminuíram mais rapidamente com DAAs, que não alcançou após 4 semanas com interferon peguilado. A resposta virológica sustentada aparece em DAAs, e a taxa de recorrência é alta na terapia com interferon em comparação com DAAs. Portanto, a reinfecção tem implicações para a eficiência do tratamento correto e para selecionar estratégias para casos de retratamento.

Hepatitis C virus genotype diversity in Ningxia Hui autonomous region, Northwestern China / Diversidade genética do vírus da hepatite C na região autônoma de Ningxia Hui

Hepatitis C virus (HCV) genotypes vary greatly in different regions. The aim of this study is to investigate the distribution of HCV genotypes in HCV infected patients, in Ningxia Hui Autonomous Region. Nucleic acid extraction and amplification were performed with test kits on 153 HCV infected patients serum samples. The HCV viral load was measured using reverse transcriptase PCR (RT-PCR) and HCV genotypes were determined. Among the 153 HCV-infected patients, 56 had genotype (GT)1b (36.60%), 45 had GT2a (29.40%), 23 had GT3a (15.00%), 14 had GT3b (9.20%),13 had GT6a (8.50%), 1 had GT1g (0.70%), 1 had GT6xa (0.70%). In GT1b, 21.40% were female and 78.60% were male; in GT2a, 42.20% were female and 57.80% were male;Males were most prevalent in genotypes 1b(39.30%), while female were most prevalent in genotype 2a(46.30%). Rare GT1g and GT6xa were also detected in males. The 41-50 year age group had the highest HCV prevalence of 32.00%. HCV GT1b is the predominant HCV genotype in Ningxia Hui Autonomous Region.

Os genótipos do vírus da hepatite C (HCV) variam muito de acordo com a região. O objetivo deste estudo é investigar a distribuição do genótipo do vírus da hepatite C em pessoas infectadas pelo vírus na região autônoma de Ningxia Hui. A extração de ácido nucleico e a expansão de amostras séricas em 153 pacientes infectados com HCV foram realizadas utilizando kits de ensaio. A capacidade do vírus HCV foi medida pela reação em cadeia da polimerase retrotranscrição (RT-PCR) e o genótipo HCV foi determinado. Os genótipos (Gt) tiveram a seguinte distribuição entre os 153 casos de infecção HCV: 56 Gt 1-B (36, 60%), 45 Gt 2-A (29, 40%), 23 Gt 3-A (15, 00%), 14 Gt 3-B (9, 20%), 13 Gt 6-A (8, 50%), 1 Gt 1-G (0, 70%) e 1 Gt 6-Xa (0, 70%). Já o sexo dos indivíduos teve a seguinte distribuição: Gt 1-B, 21, 40% mulheres e 78, 60% homens; Gt 2-A, 42, 20% mulheres e 57, 80% homens. A presença de homens é mais comum no genótipo 1-B (39, 30%), enquanto as mulheres ocorrem mais comumente no genótipo 2-A (46, 30%). Gt 1-G e Gt 6-Xa raros também foram detectados em homens. A taxa de infecção com HCV para grupos etários de 41 a 50 anos é a mais alta, com 32,00%. HCV Gt 1-B é o genótipo dominante do HCV na região autônoma de Ningxia Hui.

Recurrence of hepatitis C virus after treatment with pegylated interferon and direct acting antivirals in Punjab Pakistan / Recorrência do vírus da hepatite C após tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão

Although increased response rates concomitant in hepatitis C virus but relapse after treatment is threatened. Therefore, it is terrible requirement to evaluate the response of Pegylated interferon and direct acting antivirals in Punjab Pakistan. The study was conducted to find the rate of recurrence of HCV infection after treatment with Pegylated Interferon and Direct Acting Antivirals in Punjab Pakistan. This study was conducted at Department of Pathology, Nawaz Sharif Medical College Gujrat, while treatment effects monitored in different Government and Private Hospitals of Punjab, Pakistan. Total 973 patients who administered the recommended dose and divided in two groups (i) Interferon based therapy (ii) direct acting antivirals (DAAs).Other parameters like ALT and viral load studied. The rate of recurrence was higher in female infected with genotype 2b and in male with mixed genotype 3a/2b after six month of antiviral therapy. Genotype 3a showed significant response to therapy after three month. 32 among 374 (8.5%) were positive after 24 weeks of treatment with interferon, 29 (7.7%) patients have same genotype while 3 patients were re-infected with different HCV strains. With DAAs, only 27 (4.8%) patients were positive among 558 after 2 weeks and one patient re-infected with different genotype. Early and sustained virological response noted in DAAs. ALT and viral load decreased faster with DAAs that not achieved after 4 weeks with pegylated interferon. Sustained virological response appears in DAAs and recurrence rate is high in interferon therapy compared to DAAs. Therefore, reinfection has implications for correct treatment efficiency and to select strategies for retreatment cases.(AU)

Embora aumentem as taxas de resposta concomitantes no vírus da hepatite C (HCV), há risco de recidiva após o tratamento. Portanto, é um requisito terrível avaliar a resposta do interferon peguilado e antivirais de ação direta em Punjab, Paquistão. O estudo foi conduzido para encontrar a taxa de recorrência da infecção por HCV após o tratamento com interferon peguilado e antivirais de ação direta em Punjab, Paquistão. Este estudo foi conduzido no Departamento de Patologia Nawaz Sharif Medical College Gujrat, enquanto os efeitos do tratamento foram monitorados em diferentes hospitais públicos e privados de Punjab, Paquistão. Total de 973 pacientes que administraram a dose recomendada foram divididos em dois grupos: (i) Terapia baseada em interferon, (ii) antivirais de ação direta (DAAs). Outros parâmetros como ALT e carga viral foram estudados. A taxa de recorrência foi maior em mulheres infectadas com o genótipo 2b e em homens com genótipo misto 3a / 2b após seis meses de terapia antiviral. O genótipo 3a mostrou resposta significativa à terapia após três meses. 32 entre 374 (8,5%) foram positivos após 24 semanas de tratamento com interferon, 29 (7,7%) pacientes têm o mesmo genótipo, enquanto 3 pacientes foram reinfectados com diferentes cepas de HCV. Com DAAs, apenas 27 (4,8%) pacientes foram positivos entre 558 após duas semanas e um paciente reinfectado com genótipo diferente. Resposta virológica precoce e sustentada observada em DAAs. ALT e carga viral diminuíram mais rapidamente com DAAs, que não alcançou após 4 semanas com interferon peguilado. A resposta virológica sustentada aparece em DAAs, e a taxa de recorrência é alta na terapia com interferon em comparação com DAAs. Portanto, a reinfecção tem implicações para a eficiência do tratamento correto e para selecionar estratégias para casos de retratamento.(AU)

Virucidal activity of oriental hornet Vespa orientalis venom against hepatitis C virus

Background Hepatitis C virus (HCV) infection is a major worldwide health problem that can cause liver fibrosis and hepatocellular carcinoma (HCC). The clinical treatment of HCV infection mainly relies on the use of direct-acting antivirals (DAAs) that are usually expensive and have side effects. Therefore, achieving the discovery of more successful agents is always urgent. In this context, antiviral compounds that inhibit viral infections and disease progression with important therapeutic activities have been identified in animal venoms including arthropod toxins. This indicates that arthropod venoms represent a good natural source of promising candidates for new antivirals. Methods The antiviral activity of the wasp venom (WV), isolated from the Oriental hornet (Vespa orientalis), was assessed using cell culture technique with human hepatocellular carcinoma-derived cell line (Huh7it-1) and the recombinant strain of HCV genotype 2a (JFH1). Results The results revealed that WV inhibited HCV infectivity with 50% inhibitory concentration (IC50) of 10 ng/mL, while the 50% cytotoxic concentration (CC50) was 11,000 ng/mL. Time of addition experiment showed that the WV blocked HCV attachment/entry to the cells probably through virucidal effect. On the other hand, the venom showed no inhibitory effect on HCV replication. Conclusion WV can inhibit the entry stage of HCV infection at non-cytotoxic concentrations. Therefore, it could be considered a potential candidate for characterization of natural anti-HCV agents targeting the entry step.(AU)

Virucidal activity of oriental hornet Vespa orientalis venom against hepatitis C virus

Abstract Background Hepatitis C virus (HCV) infection is a major worldwide health problem that can cause liver fibrosis and hepatocellular carcinoma (HCC). The clinical treatment of HCV infection mainly relies on the use of direct-acting antivirals (DAAs) that are usually expensive and have side effects. Therefore, achieving the discovery of more successful agents is always urgent. In this context, antiviral compounds that inhibit viral infections and disease progression with important therapeutic activities have been identified in animal venoms including arthropod toxins. This indicates that arthropod venoms represent a good natural source of promising candidates for new antivirals. Methods The antiviral activity of the wasp venom (WV), isolated from the Oriental hornet (Vespa orientalis), was assessed using cell culture technique with human hepatocellular carcinoma-derived cell line (Huh7it-1) and the recombinant strain of HCV genotype 2a (JFH1). Results The results revealed that WV inhibited HCV infectivity with 50% inhibitory concentration (IC50) of 10 ng/mL, while the 50% cytotoxic concentration (CC50) was 11,000 ng/mL. Time of addition experiment showed that the WV blocked HCV attachment/entry to the cells probably through virucidal effect. On the other hand, the venom showed no inhibitory effect on HCV replication. Conclusion WV can inhibit the entry stage of HCV infection at non-cytotoxic concentrations. Therefore, it could be considered a potential candidate for characterization of natural anti-HCV agents targeting the entry step.

Aplicações de Metagenômica Viral em Medicina Veterinária: Análise de Viromas

As novas abordagens metagenômicas virais permitem a detecção imparcial de uma ampla gama de agentes infecciosos de maneira independente da cultura, facilitando o diagnóstico e, consequentemente, o monitoramento de doenças, o que está intimamente atrelado ao conceito One World, One Health. Além disso, abrem possibilidades para análises genéticas comparativas e enriquecimento de bancos de dados genômicos. O gênero Hepacivirus (família Flaviviridae) é um exemplo expressivo de gênero viral que cresceu rapidamente com o advento da metagenômica. Desde 1996, quando o gênero Hepacivirus foi criado, o vírus da hepatite C (HCV) era a única espécie conhecida. No entanto, os hepacivírus (HVs) têm sido detectados em diversos animais domésticos e selvagens, incluindo, equinos, cães, roedores, morcegos, bovinos, tubarões, entre outros. O objetivo geral desta tese foi aprofundar o conhecimento acerca da ecologia viral em diferentes hospedeiros, através da metagenômica na ciência veterinária, destacando diferentes subáreas em que a mesma pode ser empregada. O Capitulo 1 aborda a detecção de um hepacivírus bovino (HNV) através da plataforma de sequenciamento de alto desempenho, Illumina MiSeq. As análises revelaram uma sequência com alta divergência nucleotídica quando comparada aos demais HNVs conhecidos mundialmente. Além disso, baseado na classificação proposta na literatura, trata-se de um provável novo genótipo. Esse estudo, intitulado Highly Divergent Cattle Hepacivirus N in Southern Brazil foi publicado em 2019, na revista científica Archives of Virology. O Capítulo 2 trata-se de dois estudos de metagenômica realizados para análise de viroma. O primeiro intitulado "New Polyomavirus in Nutria, Myocastor coypus Polyomavirus 1" no qual obtivemos o sequenciamento de genoma completo de uma nova espécie de PyVs em ratão-do-banhado, pertencente ao gênero Alphapolyomavirus, que foi publicado na revista científica Archives of Virology. No segundo estudo, "Liver Virome of Healthy Pigs Reveals Diverse Small ssDNA Viral Genomes", foi analisado o viroma de figados de suínos de um abatedouro. Nesse estudo detectamos, pela primeira vez no Brasil, a presença de alguns vírus como Porcine Circovirus 1 e Porcine Parvovirus 6 e 7, além da ausência de vírus potencialmente zoonóticos. A pesquisa foi publicada na revista científica Infection, Genetics and Evolution em 2020. Concluindo, a metagenômica viral foi aplicada com sucesso na investigação de três importantes estudos para a ciência veterinária. Os resultados contribuíram com a expansão do conhecimento na área, através da descrição e caracterização de novos e conhecidos vírus, além de enriquecer os bancos de dados genômicos, provendo informação para futuras pesquisas

The viral metagenomic approaches, a culture-independent technique, allow the impartial detection of a wide range of pathogens, improving the diagnosis and, consequently, the monitoring of diseases, which is closely linked to the One World, One Health concept. Furthermore, open possibilities for comparative genetic analysis and genomic databases enrichment. Hepacivirus genus (family Flaviviridae) is an expressive example of a viral genus that grew with the advent of metagenomics. Since 1996, when the genus Hepacivirus was created, the hepatitis C virus (HCV) was the only known species. However, hepaciviruses (HVs) have been detected in several domestic and wild animals, including, horses, dogs, rodents, bats, cattle, sharks, among others. The main objective of this thesis was improving the knowledge of different hosts viral ecology through metagenomics in veterinary science, highlighting different sub-areas in which it can be applied. Chapter 1 addresses bovine hepacivirus (HNV) detection through the high-throughput sequencing (HTS), using Illumina MiSeq. The analyzes revealed a sequence with high nucleotide divergence when compared to HNVs worldwide known. In addition, based on the ICTV classification, it is a putative new genotype. This study, entitled Highly Divergent Cattle Hepacivirus N in Southern Brazil was published in 2019, in the scientific journal Archives of Virology. Chapter 2 is about two metagenomics studies performed for virome analysis. The first one entitled "New Polyomavirus in Nutria, Myocastor coypus Polyomavirus 1" in which we obtained a new PyVs complete genome species of in nutria, belonging to the genus Alphapolyomavirus, which was published in the scientific journal Archives of Virology. In the second research, "Liver Virome of Healthy Pigs Reveals Diverse Small ssDNA Viral Genomes", the swine liver virome of a slaughterhouse was analyzed. In this study, we detected, for the first time in Brazil, the presence of some viruses such as Porcine Circovirus 1 and Porcine Parvovirus 6 and 7, in addition to the absence of potentially zoonotic viruses. In conclusion, viral metagenomics has been successfully applied in important veterinary science studies. The results contributed to the knowledge expansion in the area, through the description and characterization of new and known viruses, in addition to enriching the genomic databases, providing information for future research.

Platelets can be a biological compartment for the Hepatitis C Virus

Although HCV has hepatic tropism, the presence of the virus in extra-hepatic compartments has been well documented. Platelets have been described as carriers of the virus in the circulation and may be a natural reservoir for the virus. However, few studies have been performed to evaluate the levels of HCV RNA in plasma and platelets are equal or differ in some way. Therefore, the aim of this study was to perform a comparative evaluation of the stability of HCV RNA in plasma and isolated platelets. Four aliquots of whole plasma obtained from patients infected with HCV were incubated at 37 °C for 0, 48, 96 and 144 h. After incubation, the plasma and platelet pellet was obtained from each aliquot. Viral RNA in plasma and platelets was quantified by q-PCR. The results showed a decrease in HCV RNA levels in plasma with incubation time. However, platelet HCV RNA levels were stable up to 144 h incubation. The results of this study showed that HCV RNA in platelets, although at lower concentrations than in plasma, is preserved from degradation over time, suggesting that the virus may persist longer in the body when associated with platelets, which could have an impact on the efficiency of antiviral therapy.(AU)

Seroprevalence of Hepatitis B and C markers at the population level in the municipality of Caxias do Sul, southern Brazil

Chronic viral hepatitis are main public health problems worldwide. Data about the seroprevalence to Hepatitis B and C viruses (HBV e HCV) at the population level are scarce on Brazil and especially for the Southern region of the country. The seroprevalence to HBV and HBC antigens was evaluated on a large portion of the population of the municipality of Caxias do Sul (427,858 inhabitants), Brazil. A total of 60,604 individual serum samples collected from 2008 to 2011 were screened for HBV surface antigen (HBsAg) and for antibodies against HBsAg; anti HCV antibodies were measured by ELISA (Enzyme Linked Immunosorbent Assay). Overall, 1.63% of the individuals were positive for HBsAg and 1.43% showed seropositivity to HCV. From the total, 31,749 samples were analyzed for HBsAg and 28,855 for HCV. For HBsAg, 519 samples showed positive (1.63%) while the results for 37 patients (0.12%) remained inconclusive. For the anti-HCV test, 412 individuals (1.43%) showed positive. From the positive samples for HBsAg 216 (50.6%) were from male individuals whereas for anti-HCV the seroprevalence was slightly higher for females 216 (52.4%). The higher prevalence for both hepatitis viruses were found among individuals at the age group of 40 to 59 years and the lower levels of positivity for both HBV and HBC were among children and teenagers.

Seroprevalence of Hepatitis B and C markers at the population level in the municipality of Caxias do Sul, southern Brazil

Chronic viral hepatitis are main public health problems worldwide. Data about the seroprevalence to Hepatitis B and C viruses (HBV e HCV) at the population level are scarce on Brazil and especially for the Southern region of the country. The seroprevalence to HBV and HBC antigens was evaluated on a large portion of the population of the municipality of Caxias do Sul (427,858 inhabitants), Brazil. A total of 60,604 individual serum samples collected from 2008 to 2011 were screened for HBV surface antigen (HBsAg) and for antibodies against HBsAg; anti HCV antibodies were measured by ELISA (Enzyme Linked Immunosorbent Assay). Overall, 1.63% of the individuals were positive for HBsAg and 1.43% showed seropositivity to HCV. From the total, 31,749 samples were analyzed for HBsAg and 28,855 for HCV. For HBsAg, 519 samples showed positive (1.63%) while the results for 37 patients (0.12%) remained inconclusive. For the anti-HCV test, 412 individuals (1.43%) showed positive. From the positive samples for HBsAg 216 (50.6%) were from male individuals whereas for anti-HCV the seroprevalence was slightly higher for females 216 (52.4%). The higher prevalence for both hepatitis viruses were found among individuals at the age group of 40 to 59 years and the lower levels of positivity for both HBV and HBC were among children and teenagers.

Study of anticardiolipin antibody in hepatitis C virus-positive patients

Several antibodies, including anticardiolipin antibodies (ACA), have been detected among chronically infected hepatitis C virus (HCV) patients. The present work aimed at ascertaining the clinical significance of ACA levels among HCV infection associated with two commonly encountered diseases, thrombocytopenia and arteriovenous-shunt malfunction. Six groups were studied, 11 HCV-positive thrombocytopenic patients (group I), 14 HCV-positive non-thrombocytopenic patients (group II) and 15 healthy controls (group III), 11 anti-HCV-positive hemodialysis patients with non-functioning shunt (group IV), 14 anti-HCV-positive hemodialysis patients with patent shunt (group V) (Bain Medical Equipment Co., China) and 15 healthy controls (group VI). Anticardiolipin antibody (ACA) assay was performed on all patients and controls whereas tumor necrosis factor α (TNF-α) assay was carried out on thrombocytopenic patients and controls. Thrombocytopenic groups presented an inverse correlation between IgG ACA levels and both thrombocytopenia and TNF-α levels. During the follow-up period, no other clinical manifestations related to ACA were developed. Hemodialysis groups showed a significant elevation in IgG ACA levels in groups IV and V compared to the controls, with statistically higher levels in group IV than group V. Three group IV patients were hypercholesterolemic. We can conclude that induction of proinflammatory cytokines such as TNF-α by persistent HCV infection may promote the generation of ACA. Complications of HCV, including thrombocytopenia and thrombosis in arteriovenous shunt, are more strongly correlated with IgG ACA than with IgM ACA.(AU)

Study of anticardiolipin antibody in hepatitis C virus-positive patients

Several antibodies, including anticardiolipin antibodies (ACA), have been detected among chronically infected hepatitis C virus (HCV) patients. The present work aimed at ascertaining the clinical significance of ACA levels among HCV infection associated with two commonly encountered diseases, thrombocytopenia and arteriovenous-shunt malfunction. Six groups were studied, 11 HCV-positive thrombocytopenic patients (group I), 14 HCV-positive non-thrombocytopenic patients (group II) and 15 healthy controls (group III), 11 anti-HCV-positive hemodialysis patients with non-functioning shunt (group IV), 14 anti-HCV-positive hemodialysis patients with patent shunt (group V) (Bain Medical Equipment Co., China) and 15 healthy controls (group VI). Anticardiolipin antibody (ACA) assay was performed on all patients and controls whereas tumor necrosis factor α (TNF-α) assay was carried out on thrombocytopenic patients and controls. Thrombocytopenic groups presented an inverse correlation between IgG ACA levels and both thrombocytopenia and TNF-α levels. During the follow-up period, no other clinical manifestations related to ACA were developed. Hemodialysis groups showed a significant elevation in IgG ACA levels in groups IV and V compared to the controls, with statistically higher levels in group IV than group V. Three group IV patients were hypercholesterolemic. We can conclude that induction of proinflammatory cytokines such as TNF-α by persistent HCV infection may promote the generation of ACA. Complications of HCV, including thrombocytopenia and thrombosis in arteriovenous shunt, are more strongly correlated with IgG ACA than with IgM ACA.(AU)

Toxoplasma gondii em pacientes em tratamento para hepatite C, no sul do Brasil

Toxoplasma gondii é um protozoário oportunista causador de infecções em indivíduos com o sistema imunológico comprometido. Este estudo visa conhecer a soroprevalência para T. gondii em pacientes submetidos a tratamento para hepatite C em Centro de Referência no sul do Rio Grande do Sul, os possíveis efeitos desse tratamento na imunidade e os fatores de risco para a infecção pelo parasito. Segundo o conhecimento dos autores, esse foi o primeiro trabalho com essa categoria de pacientes. Foi realizado um estudo transversal prospectivo, com um total de 57 pacientes, os quais foram acompanhados antes e durante o tratamento, num período de 24 ou 48 semanas, dependendo do genótipo e da resposta virológica. A pesquisa sorológica de anticorpos do tipo IgG foi realizada através da técnica de Imunofluorescência Indireta (IFI) e os anticorpos IgM foram detectados por Eletroquimioluminescência (ECLIA). Os dados epidemiológicos foram obtidos pela aplicação de um questionário e os clínicos através da análise dos prontuários dos pacientes, previamente autorizados. A soroprevalência para T. gondii foi de 75,4%. A soropositividade foi significativamente maior entre os pacientes com hábito de mexer na terra e/ou areia (OR = 14,84) e com falta de conhecimento sobre a infecção (OR =12,98). Embora não tenha ocorrido nenhum caso de reagudização da doença, a elevada soropositividade e o desconhecimento constatados indicam a necessidade de monitoramento dos pacientes em tratamento para hepatite C, uma vez que essa infecção e os fármacos utilizados podem comprometer a resposta imunológica dos pacientes.

Toxoplasma gondii is an opportunistic protozoan which causes infections in individuals with the imune system compromised. This study aims to know the soroprevalence of T. gondii in patients submitted to the hepatitis C treatment in reference center in the south of Rio Grande do Sul, the probable effects of this kind of treatment in the immunity and the risk factors to the infection by this parasite. According to the knowledge from the authors, this was the first study with this patients category. It was done a prospective transversal study, with the total number of 57 patients who were monitored before and during the treatment in 24 or 48 week period, depending on the genotype and the virologic response. The antibody serological research of IgG type was done by the Indirect Immunofluorescence (IFI) technique and the IgM antibodies were detected by Electrochemiluminescense (ECLIA). The epidemiological data were obtained by a questionnaire application and the clinic data by analyzing the patients records, previously permitted. The soroprevalence to T. gondii was 75.4%. The seropositivity was significantly larger among the patients who have the habit to deal with the soil and/or sand (OR = 14,84) and with the lack of knowledge about the infection (OR =12,98). Although it doesnt happened any case of illness reaggravating, the high seropositivity and the lack of knowledge which were noticed, demonstrate the necessity of monitoring patients under the treatment to the hepatitis C, once this infection and the drugs used can compromise the patients immunological response.

Distribution of Hepatitis C virus (HCV) genotypes in seropositive patients in the state of Alagoas, Brazil

We determined the frequency of hepatitis C virus (HCV) genotypes in anti-HCV seropositive patients in the state of Alagoas, Brazil, by means of nested-reverse transcription-polymerase chain reaction (RT-nested-PCR) followed by restriction fragment length polymorphism (RFLP) of amplified fragments of the 5´NCR. The nested-PCR with genotype-specific primers from the core region was carried out when detection was not possible by the first approach. Detectable HCV-RNA was present in 115 (74.7%) of 154 serum samples. Genotype 1 was the most frequent (77.4%), against 20.9% of genotype 3 and 0.8% of genotype 2. Subtype 1b was predominant (65.2%), followed by subtypes 1a (8.7%), and 3a (6.1%). Coinfection (1a/3a) was detected in 0.8% of the samples. Indeed, there was no significant differences in the prevalence of genotype 1 compared to what has been obtained from anti-HCV seropositive patients from other locations in Brazil. Here we report for the first time the genotype 2 in the state of Alagoas.

A frequência de genótipos do vírus da hepatite C (HCV) em pacientes soropositivos anti-HCV no estado de Alagoas, Brasil, foi determinada através da RT-PCR aninhada da região 5'NCR seguida pela análise do polimorfismo de comprimento dos fragmentos de restrição (RFLP). A RT-PCR aninhada utilizando primers genótipo-específicos da região core foi efetuada quando não foi possível determinar o genótipo pelo primeiro método. Níveis detectáveis de HCV-RNA estavam presentes em 115 (74,7%) das 154 amostras de soro. O genótipo 1 foi o mais freqüente (77,4%), contra 20,9% do genótipo 3 e 0,8% do genótipo 2. O subtipo 1b foi predominante (65,2%), seguido pelos subtipos 1a (8,7%) e 3a (6,1%). Co-infecção (1a/3a) foi detectada em 0,8% das amostras. Não foram encontradas diferenças significativas quanto à prevalência do genótipo 1 em relação ao que tem sido obtido de pacientes soropositivos anti-HCV de outras localidades do Brasil. Este é o primeiro relato da presença do genótipo 2 no estado.

Viral Hepatitis C in a leading Brazilian hospital: epidemiological factors and genotyping

The hepatitis C virus (HCV) is classified into six different genotypes and their distribution is different throughout the world. Epidemiologic studies are important to determine several characteristics of the virus, as well as the disease. This study analysed the prevalence of HCV and its genotypes among patients from a leading hospital in Ceará, which is located in Northeast Brazil. A total of 119 anti-HCV-seropositive patients, each having previously completed a questionnaire about risk behaviours related to HCV infection were tested for HCV infection using a qualitative HCV polymerase chain reaction (PCR) assay and genotyping by restriction fragment length polymorphism (RFLP). The detection was based on amplifying of the non-coding 5' region. Of the 119 patients, 95 showed positive results in the qualitative HCV test. History of surgery was the most reported risk factor, followed by the use of drugs, having tattoos, undergoing haemodialysis and occupational exposure. Genotype 1 was the most prevalent (46.9%), followed by genotype 3 (34.4%) and 2 (8.3%). The genotype distribution was similar for all of the various risk behaviours.

O vírus da hepatite C (VHC) é classificado em seis genótipos diferentes e sua distribuição é diferente em todo o mundo. Os estudos epidemiológicos são importantes para determinar várias características sobre o vírus, bem como da doença. Este estudo analisou a prevalência do VHC e seus genótipos em pacientes atendidos em hospital de referência no Ceará, o qual é localizado no nordeste do Brasil. Um total de 119 pacientes, os quais eram soropositivos anti-VHC, preencheram questionários sobre fatores de risco relacionados à infecção pelo VHC e foram testados quanto à infecção ao VHC usando o teste da reação da polimerase em cadeia (PCR) qualitativo para VHC e genotipagem por "restriction fragment length polymorphism" (RFLP). A detecção foi baseada na amplificação da região não codificante 5'. Dos 119 pacientes, 95 mostraram resultados positivos no teste qualitativo para VHC. A história prévia de cirurgia foi o fator de risco mais relatado, seguido pelo uso de drogas, ter tatuagem, ter sido submetido à hemodiálise e risco ocupacional. O genótipo 1 foi o mais prevalente (46,9%), seguido pelo genótipo 3 (34,4%) e 2 (8,3%). A distribuição dos genótipos foi similar entre os vários fatores de risco analisados.