Takao Shibayama

Prolonged Fecal Shedding of Hepatitis E Virus (HEV) during Sporadic Acute Hepatitis E: Evaluation of Infectivity of HEV in Fecal Specimens in a Cell Culture System

ABSTRACT To investigate the duration of fecal shedding and changing loads of hepatitis E virus (HEV) in feces and serum from patients with acute HEV infection, HEV RNA was quantitated in periodic serum and fecal specimens obtained from 11 patients with sporadic acute hepatitis E. All 11 patients had detectable HEV RNA in serum at admission, with the highest viral load being 1.9 × 103 to 1.7 × 107 copies/ml, and HEV viremia lasted until days 17 to 48 (mean, 28.3) after the onset of hepatitis. Even at the initial examination on days 10 to 29 (mean, 17.6), the HEV load in fecal supernatant was less than 5.7 × 104 copies/ml for 10 of the 11 patients, while for the remaining patient (patient 1) it was markedly high, 2.0 × 107 copies/ml on day 22. In addition, although HEV RNA in fecal supernatant continued to be positive until days 14 to 33 (mean, 22.4) for patients 2 to 11, that for patient 1 was detectable even on day 121. HEVs in fecal specimens obtained on days 22, 24, 26, 28, and 30, but not day 121, from patient 1 grew efficiently in PLC/PRF/5 cells, reaching the highest titer of up to 107 copies/ml in culture medium on day 50 postinoculation. The HEV genome recovered from patient 1 had 29 unique nucleotides that were not seen in any of the 25 reported HEV isolates of the same genotype over the entire genome, with six amino acid substitutions in the ORF1 protein.

Inverse relationship between the titre of TT virus DNA and the CD4 cell count in patients infected with HIV.

ObjectivesTo investigate the prevalence and relative titre of TT virus (TTV) DNA, and to examine the relationship between the extent of TTV viraemia and the immune status among 144 patients with HIV infection; 178 age- and sex-matched healthy individuals were also studied. MethodsTTV DNA was detected quantitatively by two distinct polymerase chain reaction (PCR) methods [untranslated region (UTR) and N22]. UTR PCR detects all TTV genotypes, and N22 PCR can primarily detect four major TTV genotypes (1–4). ResultsUsing UTR PCR and N22 PCR, respectively, TTV DNA was detected significantly more frequently in HIV-infected patients than in controls (99 versus 91%, P < 0.001; 56 versus 27%, P < 0.0001), and the relative titre (10N/ml) was significantly higher in HIV-infected patients [4.5 ± 1.2 (mean ± SD) versus 3.1 ± 0.9, P < 0.0001; 2.6 ± 1.5 versus 1.5 ± 0.9, P < 0.0001]. Age, sex, co-infection with hepatitis B or C virus, and risk factors for HIV transmission did not appear to be significant factors associated with the titre of TTV viraemia. However, the titre of TTV DNA was significantly higher in HIV-infected patients with AIDS (P < 0.0001), those with low CD4 T cell count (P < 0.0001), or those with high HIV viral loads (P = 0.0047). ConclusionTTV is highly prevalent and high-titred in HIV-infected patients. The TTV viral load may reflect the degree of immune status of these immunocompromised hosts.

Localization of hepatitis A virus in marmoset liver tissue during the acute phase of experimental infection

SummaryElectron microscopic and virological studies of marmoset liver tissue with acute infection of hepatitis A virus (HAV), especially in the earlier stages of infection, were carried out to characterize the maturation process of HAV. Four marmosets were inoculated intravenously with HAV suspension and sacrificed 1 week, 2 weeks, 3 weeks and 4 weeks after inoculation respectively. Hepatitis A antigen (HAAg) in 10% liver homogenates of marmosets was examined by radioimmunoassay and a large amount of HAAg was detected in the liver homogenate of two marmosets sacrificed 2 weeks and 3 weeks after inoculation respectively. The histodiagnosis of the marmoset sacrificed 2 weeks after HAV inoculation was normal. However, many clusters of virus-like particles about 27 nm in diameter, in both “solid” and “empty” forms were found, mainly in vesicles of Kupffer cells by electron microscopy. In the animal that developed mild hepatitis 3 weeks after inoculation HAV-like particles were found in vesicles of hepatocytes by electron microscopy. By immune electron microscopy using peroxidase-conjugated anti-hepatitis A antibody, HAAg was detected on the particles present within the cytoplasmic vesicles of Kupffer cells or hepatocytes and on the surrounding membrane of the vesicles which contained HAV-like particles.

Infection with GB virus C in the patients with primary hepatocellular carcinoma in Japan

Abstract RNA of recently reported, putative non-A to E hepatitis virus designated GB virus C (GBV-C) was determined by reverse-transcription polymerase chain reaction in sera from 231 Japanese patients with primary hepatocellular carcinoma. GBV-C RNA was detected in 21 patients (9% of the total), including two of the 23 patients (8%) with markers of hepatitis B virus infection, 11 of the 114 patients (10%) with markers of hepatitis C virus infection, seven of the 86 patients (8%) with markers of both hepatitis B and C virus infections, and one of the eight patients (13%) without such markers. These results indicate that the contribution of GBV-C infection to the development of hepatocellular carcinoma would be small either by itself or in cooperation with hepatitis B and C viruses.

Risk Factors of Hepatocellular Carcinoma in Chronic Hepatitis C and Cirrhosis: Special Reference to Laparoscopic Findings

Abstract: It remains unclear whether the hepatitis C virus genotype is associated with the severity and outcome of HCV‐related liver disease. The aim of this study was to determine whether hepatitis C virus genotype influenced the risk of developing hepatocelMar carcinoma. Two hundred and sixty nine patients who had chronic hepatitis C and cirrhosis without hepatocellular carcinoma were studied. The stage and activity of hepatitis were determined at laparoscopy and patients were followed up until the development of hepatocellular carcinoma or for a maximum of 16 years. Hepatitis C virus genotypes were determined by a genotyping enzyme‐linked immunosorbent assay. A cross‐sectional study revealed that the prevalence of hepatitis C virus genotype 1 increased and that of genotype 2 decreased with the progression of liver disease (pc 0.01). A follow‐up study using the Kaplan‐Meier method showed that hepatocellular carcinoma occurred more frequently in patients with hepatitis C virus genotype 1 (p<0.01), patients with a more advanced disease stage (p<0.01), and patients with reddish markings (p<0.05). Cox multivariate proportional hazards analysis confirmed that these three risk factors were independent. Hepatocellular carcinoma developed more frequently in patients with hepatitis C virus genotype 1 and pre‐cirrhosis (stage 3 chronic hepatitis with nodules) or liver cirrhosis, in whom hepatitis showed continued activity and progression. (Dig Endosc 1999; 11: 24–31)

FORMATION AND CLINICAL SIGNIFICANCE OF REDDISH MARKINGS ON THE LIVER SURFACE : ACTIVITY OF CHRONIC LIVER DISEASE

Background:  The aim of this study was to clarify the clinical significance of reddish markings appearing on the surface of the liver.