Hepatitis C virus RNA and hepatitis C virus antibody in the serum of patients with abnormal liver function.

In order to elucidate the relation between hepatitis C virus (HCV) RNA and antibody to HCV (anti-HCV) in serum, we examined samples of serum collected from 228 HBsAg-negative patients, with abnormal alanine aminotransferase (ALT) values, for HCV-RNA by nested polymerase chain reaction (PCR) assay and for anti-HCV using C100 protein as the antigen. HCV-RNA was detected in 99 (92.5%) of 107 anti-HCV-IgG-positive samples, regardless of ELISA optical density cut-off value (ELISA ratio), and in 34 (28.1%) of 121 anti-HCV-IgG-negative samples in which the frequency of the presence of HCV-RNA became higher in proportion to the ELISA ratio. Among 42 discordant cases (34 anti-HCV-IgG-negative, RNA-positive cases and eight anti-HCV-IgG-positive, RNA-negative cases), 10 were positive for anti-HCV-IgM (8/34 and 2/8, respectively) irrespective of clinical status. These findings suggest that in patients with abnormal ALT values, even if they are anti-HCV-IgG negative, HCV infection cannot be excluded. Furthermore, PCR assay for detecting HCV-RNA may be more suitable for identifying patients with infectious virus than is detection of anti-HCV-IgG. Detection of anti-HCV-IgM may also be useful.

Prevalence of blood-borne viruses among intravenous drug users and alcoholics in Hiroshima, Japan.

We investigated the prevalence of human immunodeficiency viruses-1 and 2 (HIV-1 and HIV-2), human T-lymphotropic virus type I and II, hepatitis B virus (HBV), hepatitis C virus (HCV) and hepatitis D virus among intravenous drug users (IVDU) in Hiroshima, Japan, where little is known about their present levels. From June to December 1993, serum samples were collected from 47 IVDU and 98 alcoholics in Hiroshima, Japan, and examined for markers of virus infection. The prevalence of antibody to HCV (anti-HCV) and/or HCV-RNA was significantly higher in IVDU than alcoholics (74.5% vs 20.4%, 44.7% vs 10.2% respectively, P < 0.001). In contrast, the prevalence of antibody to hepatitis B surface antigen and/or core antigen (anti-HBs and/or anti-HBc) showed no significant difference between the 2 groups (57.4% vs 66.3%). HIV-1 infection was found in one (2.1%) IVDU and genome analysis indicated that it was subtype B according to Myers' classification. Thus, an extremely low level of HIV infection and a high level of HCV infection was found in IVDU. Careful follow-up of this group is thought to be needed to minimize an outbreak of HIV-1 infection in Japan.

A long-term follow-up of five patients with hepatitis delta virus in Japan.

From 1973 to 1989 five patients with hepatitis delta virus having anti-hepatitis delta antibodies continuously in the serum for more than 5 years were identified among 1019 hepatitis B virus carriers who were being followed-up for more than 3 years (mean 8.9 years). Of the five patients with antibodies, three had a history of blood transfusion, in two cases the transfusion was massive, and one patient had been addicted to narcotics given intravenously 35 years before. In the remaining patient, the route of superinfection could not be determined. Hepatitis delta antigen was detected in hepatocyte nuclei of one of the three patients in whom liver biopsies were performed and there was chronic persistent hepatitis detected by an indirect immunoperoxidase technique. During the follow-up, hepatocellular carcinoma developed in one case but the clinical prognosis was favourable in the remaining four cases.

Persistence of hepatitis B e antigen for a long period after the clearance of hepatitis B surface antigen.

Four-hundred and seventy-eight hepatitis B virus (HBV) carriers were followed for more than 60 months. In thirty-six cases the hepatitis B surface antigen (HBsAg) was cleared from serum during the observation period. In one case hepatitis B e antigen (HBeAg) was detectable for more than 66 months even after the appearance of antibody against hepatitis B surface antigen (anti-HBs). HBeAg was detected not only by radioimmunoassay (RIA) and enzyme-linked immunosorbent assay (ELISA), but also by reversed passive haemagglutination (RPHA) method which has been believed to be far less sensitive than RIA or ELISA tests. HBV DNA was demonstrated in the serum even after the appearance of anti-HBs. Therefore, it is suggested that HBV (Dane particles) remains or reappears intermittently in the serum after seroconversion of HBsAg antigenaemia to anti-HBs complexes.

[Detection of hepatitis delta virus (HDV) markers in HBV carriers].

To investigate the prevalence and clinical features of hepatitis delta virus (HDV) superinfection in hepatitis B virus (HBV) carriers, antibody to hepatitis delta antigen (anti-HD) was determined in the sera of 328 HBV carriers in Japan. 1) Of the 328 HBV carriers, six (1.8%) were seropositive for anti-HD by enzyme linked immunosorbent assay. IgM-antibody to hepatitis delta antigen was detected in 2/6 patients with a high anti-HDV titer. None of the patients was positive for hepatitis dealt antigen. 2) HBV carriers with chronic liver disease had a greater frequency of seropositivity of anti-HD than asymptomatic HBV carriers. These data indicate that HDV superinfection may be an etiologic agent of chronic liver disease in HBV carriers.

[Studies on hepatitis C virus infection in haemodialysis patients].

To examine the prevalence of hepatitis C virus (HCV) in haemodialysis patients without blood transfusion in Hiroshima Prefecture, antibody to HCV (anti-HCV) was studied by the Ortho ELISA Kit in sera from 393 consecutive haemodialysis patients and in sera from 510 age and sex matched healthy members of the general population (control). An additional confirmatory test was done by a recombinant immunoblot assay. 1) Anti-HCV was detected in 70 of the 393 dialysis patients and 3 of the 510 healthy controls (17.8% vs 0.6%, p less than 0.01). Prevalence of anti-HCV in haemodialysis patients sera was increased by the volume of blood transfusion, and even in dialysis patients who had no blood transfusion, the frequency of anti-HCV positivity (9.2%) was greater than the healthy controls (p less than 0.01). Thus, the major route of HCV transmission in haemodialysis patients without blood transfusion may be via the haemodialysis treatment. 2) The prevalence of anti-HCV increased significantly with the ALT level and abnormal ALT activity of the anti-HCV positive group were significantly greater than that of the negative group. Thus, it is suggested that HCV infection may be an etiologic factor of liver dysfunction in haemodialysis patients.

[HBV carriers and HDV superinfection--studies on the cases of HBsAg clearance].

We studied the annual clearance rates of hepatitis B surface antigen (HBsAg) and the annual seroconversion rates of HBsAg (HBs seroconversion rates), and the correlation between HBsAg clearance and hepatitis delta virus (HDV) superinfection in hepatitis B virus (HBV) carriers in Japan. Out of 1,029 HBV carriers followed for more than 36 months, 56 cases were cleared of HBsAg from the sera, and 24 of these cases developed hepatitis B surface antibody. The annual clearance rate of HBsAg was 0.94% and the annual HBs seroconversion rate was 0.27%. These rates increased with aging, especially above 30 years of age. Antibody to HDV was detected in three cases with increased serum alanine aminotransferase activity preceding HBsAg clearance. These data indicate that HDV superinfection may play a role in induction of the HBsAg clearance in HBV carriers in Japan.

[Combination therapy with ofloxacin and chloramphenicol against typhoid fever].

Recently we experienced two male typhoid patients who required additional treatment with CP sodium succinate (CP succinate) to OFLX therapy. Although both of the cases were administered 900 mg/day of OFLX orally for three and four days, respectively, the clinical efficacies were not sufficient. We added 2.0 g/day of CP succinate intravenously for six days, although a lasting high fever returned to normal level in a few days. The administration of OFLX was continued up to 14 days. The daily doses of OFLX during the co-medication with CP succinate were 600 mg in one case and 900 mg in the other case. In both cases no recurrence was observed clinically and bacteriologically for six months after their discharge. No adverse reaction was observed throughout the therapy.

[Studies on the clinical and bacteriological effects of ofloxacin on the typhoid patients and carriers].

Fundamental and clinical studies of OFLX were performed against the patients with typhoid fever and typhoid carriers. 1) Clinical and bacteriological effects: Eight patients with typhoid fever and 3 typhoid carriers were treated with OFLX. Daily doses of the agent were 900 mg in 5 adult patients, 600 mg in a child patient and 3 adult carriers. In one case of the remaining 2 adult patients, daily doses of the agent changed from 800 mg to 1200 mg and from 900 mg to 1200 mg in the other one. The duration of the treatment was 9, 14 or 21 days. Clinical efficacies of OFLX against the patients proved 4 cases were "excellent", 3 cases were "good" and one case was "poor". The eradication of Salmonella typhi recognized in all cases containing 3 carriers with the exception of the "poor" case. Adverse reactions were observed transiently in 3 patients, such a slight decrease of RBC count, decrease of granulocyte count and elevation of GPT value respectively. 2) Antimicrobial activity: MICs of OFLX against 40 strains of S. typhi were 0.05 micrograms/ml and 0.1 micrograms/ml. The MICs of NFLX, CPFX and T-3262 were almost the same as that of OFLX, and those of ENX, NY-198 and NA were higher than that of OFLX. The peaks of MIC of CP and ABPC, first choice drug against typhoid fever, were 1.56 micrograms/ml and 0.38 micrograms/ml respectively. 3) Serum concentration; Serum concentrations of OFLX were serially measured on 5 patients through the day. The concentrations of the drug were distributed from 0.82 micrograms/ml to 6.34 micrograms/ml at 6.30 a.m. and from 2.52 micrograms/ml to 11.2 micrograms/ml at 9:00 p.m. Those of the day time showed considerable individual differences.

[Antitumor effects of cisplatin, cyclophosphamide and interferon-gamma (gamma-IFN) against argyrophil small cell carcinoma of the uterine cervix heterotransplanted into nude mice].

We investigated antitumor effect of cis-diammine-dichloroplatinum (cisplatin or CDDP), cyclophosphamide (CY) and interferon-gamma (gamma-IFN) against argyrophil small cell carcinoma (ASCC) of the uterine cervix using heterotransplanted ASCC tumor (YIK-1) into nude mice, containing HPV 16 DNA in a multicopy integrated form. No tumor growth retardation was observed in the nude mice which received the single administration of CDDP 2 mg/kg, CY 10 mg/kg or gamma-IFN 1 x 10(7) U/kg. However, the combined administration of CDDP and CY, or CDDP, CY and gamma-IFN markedly inhibited the tumor growth. Moreover, with comparison about relative tumor volume between these two groups, the combination of CDDP, CY and gamma-IFN was more effective than that of CDDP and CY. These data suggest that the combination chemotherapy of CDDP, CY and/or gamma-IFN was effective for the suppression of tumor growth in argyrophil small cell carcinoma of the uterine cervix. Further study on the antitumor potentiation of various anti-cancer agent and the combination of them against ASCC is needed.