Nitric oxide levels are not changed in saliva of patients infected with hepatitis C virus

O objetivo desta investigacão foi avaliar os níveis de metabólitos do óxido nítrico na saliva de pacientes anti-vírus da hepatite C positivos na tentativa de correlacionar os níveis desses metabólitos com a presenca do VHC na saliva. Foram estudados 39 pacientes anti-VHC positivos (9 com enzimas hepáticas normais, 16 com hepatite crônica e 14 com cirrose hepática) e em 13 controles saudáveis, sem sinais ou sintomas de doenca hepática.O RNA do VHC foi identificado no soro e na saliva através de técnica de RT-PCR e os níveis de óxido nítrico foram avaliados pela quantificacão dos seus metabólitos estáveis, nitratos e nitritos. Os resultados demonstraram que os níveis de nitrito na saliva não diferiram significativamente no grupo anti-VHC positivo em relacão ao grupo controle, nem entre os grupo com presenca ou ausência do RNA do VHC na saliva. Os níveis de nitrito foram mais elevados no grupo com cirrose hepática do que nos grupos controle e anti-VHC positivos, sem cirrose hepática, mas as diferencas não foram estatisticamente significativas. A não observacão de níveis elevados de nitrito na saliva dos pacientes anti-VHC positivos é uma indicacão indireta de que a sialoadenite não deve ser freqüente nesses pacientes ou, se existe, é de intensidade não suficiente para modificar os níves de óxido nítrico na secrecão salivar.

Detection of hepatitis C virus RNA in saliva samples from patients with seric anti-HCV antibodies.

We examined the frequency of HCV-RNA in saliva samples from anti-HCV positive patients. Both plasma and saliva samples from 39 HCV patients (13 with normal liver enzymes, 19 with abnormal liver enzymes and 13 with cirrhosis) were investigated. Stimulated saliva and fresh plasma were centrifuged (900 x g,10 min) and stored at -70 degrees C, after the addition of guanidine isothiocyanate RNA extraction buffer. HCV-RNA was detected by RT-nested-PCR (amplification of HCV-cDNA for two rounds, using HCV primers 939/209 and 940/211). HCV genotyping was carried out by RFLP (using Mva I and Hinf 1 or Hae III and Rsa I restriction enzymes). Thirty-two out of 39 (82%; 95% CI=70-94%) anti-HCV-positive patients had HCV-RNA in plasma samples. Eight out of 39 (20.5%; 95% CI=6.6-34.4%) had HCV-RNA in the saliva. The HCV genotype in saliva samples from these patients matched the genotype found for plasma HCV-RNA. No significant correlation between the presence of HCV and either age, gender, HCV genotype or any risk factor for HCV infection was found. The observed prevalence (20.5% of anti HCV positive patients or 25% of the patients with HCV-RNA in plasma) was lower than that previously reported from other countries. The low frequency of HCV-RNA in saliva samples observed in our study may be due to the use of cell-free saliva. Other authors reporting higher frequencies of HCV-RNA in saliva used whole saliva, without centrifugation.

Detection of hepatitis C virus RNA in saliva samples from patients with seric anti-HCV antibodies

We examined the frequency of HCV-RNA in saliva samples from anti-HCV positive patients. Both plasma and saliva samples from 39 HCV patients (13 with normal liver enzymes, 19 with abnormal liver enzymes and 13 with cirrhosis) were investigated. Stimulated saliva and fresh plasma were centrifuged (900 x g,10 min) and stored at -70°C, after the addition of guanidine isothiocyanate RNA extraction buffer. HCV-RNA was detected by RT- nested-PCR (amplification of HCV-cDNA for two rounds, using HCV primers 939/209 and 940/211). HCV genotyping was carried out by RFLP (using Mva I and Hinf 1 or Hae III and Rsa I restriction enzymes). Thirty-two out of 39 (82 percent; 95 percent CI=70-94 percent) anti-HCV-positive patients had HCV-RNA in plasma samples. Eight out of 39 (20.5 percent; 95 percent CI=6.6-34.4 percent) had HCV-RNA in the saliva. The HCV genotype in saliva samples from these patients matched the genotype found for plasma HCV-RNA. No significant correlation between the presence of HCV and either age, gender, HCV genotype or any risk factor for HCV infection was found. The observed prevalence (20.5 percent of anti HCV positive patients or 25 percent of the patients with HCV-RNA in plasma) was lower than that previously reported from other countries. The low frequency of HCV-RNA in saliva samples observed in our study may be due to the use of cell-free saliva. Other authors reporting higher frequencies of HCV-RNA in saliva used whole saliva, without centrifugation.

Nitric oxide levels are not changed in saliva of patients infected with hepatitis C virus.

The aim of this investigation was to determine nitric oxide metabolite levels in saliva samples from hepatitis C virus-positive patients in an attempt to test the hypothesis if increased levels of nitric oxide metabolites correlates with the presence of HCV-RNA in saliva. Saliva of 39 HCV-positive patients and 13 HCV-negative patients, without clinical or laboratorial evidence of liver disease were tested for nitric oxide metabolites. HCV-RNA was detected in serum and saliva by a RT-PCR method and nitric oxide level was determined by evaluation of its stable degradation products, nitrate and nitrite. No differences were found between the concentration of nitrite in saliva from HCV patients and controls, in despite of the presence or not of HCV RNA in saliva. Patients with HCV and cirrhosis had higher concentrations of nitrite but not significantly different from the control group or the groups of anti-HCV patients without cirrhosis. Increased levels of nitrite were not detected in anti-HCV positive patients, an indirect indication that chronic sialoadenitis are infrequent in these patients or occurs with low intensity not sufficient to increase nitric oxide metabolite levels in saliva.