Tetsuzo Koda

Risk of liver cirrhosis and hepatocellular carcinoma in subjects with hepatitis B and delta virus infection: A study from Kure, Japan

To investigate the effect of hepatitis delta virus (HDV) superinfection on the long‐term outcome of Japanese subjects with chronic hepatitis B virus (HBV) infection, we examined the presence of antibodies to hepatitis delta antigen (anti‐HD) in serial serum samples collected from 1127 subjects with chronic HBV infection. The subjects were followed for at least 36 months (mean: 121.3 months) between 1973 and 1991. Among 69 cases where anti‐HD was detected, eight (12%) developed liver cirrhosis (LC) and six (9%) developed hepatocellular carcinoma (HCC). However, among 1058 cases without anti‐HD, there were 43 patients (4%) who developed LC and 29 (3%) who developed HCC. The prevalence of LC and HCC was significantly higher among the cases with anti‐HD than those without anti‐HD. The proportion of LC and HCC per 1000 person years was 10.46 and 7.84, respectively among cases with anti‐HD, and 4.05 and 2.73 among those without anti‐HD, respectively. The overall relative risk of LC and HCC was 2.58 and 2.87, respectively; 95% confidence interval (CI): LC, 1.14–5.13; HCC, 1.03–6.23. These results indicate that in the Kure district in Japan, where HDV infection of persons infected with HBV is about 6%, such superinfection increases the risk of LC and HCC.

Application of tyramide signal amplification for detection of N-glycolylneuraminic acid in human hepatocellular carcinoma.

AbstractBackground. N-Acetylneuraminic acid and N-glycolylneuraminic acid (NeuGc) are the most common sialic acids in mammals, and NeuGc has attracted attention as a tumor-associated antigen. Methods. In frozen liver sections from patients with hepatocellular carcinoma, glycolipid-type NeuGc was detected on the surface of liver cancer cells in 9 of 17 samples (52.9%) by immunostaining, using two chicken monoclonal antibodies against NeuGc and the tyramide signal amplification method. When conventional immunostaining without amplification was used, all 17 specimens tested were negative. Results. Increased serum levels of anti-NeuGc IgG and/or IgM were observed in 13 of the17 patients with hepatocellular carcinoma (76.5%). The presence of these antibodies was mostly attributed to the expression of NeuGc on hepatocellular carcinoma cells. Of the subjects with small HCCs (diameter 3 cm or less), 6 of 10 were positive for serum anti-NeuGc antibodies; however, 1 of these was negative for both serum Α-fetoprotein (AFP) and for prothrombin–induced vitamin K antagonist II (PIVKA-II). There was no correlation between serum AFP- or PIVKA-II, levels and the presence of NeuGc or anti-NeuGc IgG and/or IgM. Conclusion. The tyramide signal amplification method is useful for the immunohistochemical detection of low-level NeuGc expression by hepatocellular carcinoma cells. We therefore consider that measurement of serum levels of anti-NeuGc antibodies is clinically meaningful and that anti-NeuGc antibody may be a useful screening test, in combination with AFP and PIVKA-II, for the early diagnosis of hepatocellular carcinoma.

Detection of the Hanganutziu-Deicher antigen in patients with hepatocellular carcinoma

Abstract The heterophilic Hanganutziu-Deicher (H-D) antigen, is a tumor-associated antigen of humans and chickens which is not detectable in normal individuals. H-D antibodies have been found in the sera of patients with hepatocellular carcinoma (HCC) and various underlying liver diseases. However, the H-D antigen has not yet been detected in liver tissue. HCC cells obtained from 15 patients were examined for the presence of the H-D antigen using two chicken anti-H-D monoclonal antibodies by flow cytometry (FCM). The H-D antigen (glycolipid type) was detected in three of the 15 HCC samples. Serum IgG and/or IgM type H-D antibodies were increased in 12 of the patients (80%). Thirteen patients were positive for either HBsAg or anti-HCV where the other two were negative for both. This suggests that hepatitis virus is involved in the expression of H-D antigen. These results indicate that increased levels of those antibodies are attributable to the expression of H-D antigen in HCC.

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.

Hepatitis δ virus infection in different time periods in Japan

To investigate the prevalence of hepatitis δ virus (HDV) infection in different time periods between 1973 and 1988, antibody to hepatitis δ antigen (anti‐HD) was tested for in sera collected from 1088 cases with acute or chronic hepatitis B virus (HBV) infection treated at Kure National Hospital. Between 1979 and 1983, anti‐HD was first detected in 16% (four of 25 cases) of patients with acute hepatitis B, in 6.8% (11 of 161 cases) of asymptomatic HBV carriers and 26% (51 of 196 cases) of those with chronic liver disease. Except for this time period anti‐HD was hardly detected. These findings indicate that sporadic HDV infections existed in this area between 1979 and 1983.

Abnormal prothrombin : evaluation as a tumour marker and localization in tissues of patients with hepatocellular carcinoma

Abstract In this study, the diagnostic significance of PIVKA‐II concentrations in various liver diseases was evaluated, and the use of PIVKA‐II as a tumour marker for hepatocellular carcinoma (HCC) was discussed. Also, the location of abnormal prothrombin (PIVKA‐II) production in HCC by indirect immunoperoxidase staining was examined. There was a good correlation between plasma and serum PIVKA‐II concentrations, indicating that serum samples are adequate for PIVKA‐II measurements. Fifty‐four of 97 (55.7%) patients with HCC, one of 10 (10%) patients with metastatic liver cancer and two of 47 (4.3%) patients with liver cirrhosis had positive serum PIVKA‐II concentrations. Positive serum PIVKA‐II concentrations were found more frequently in patients with HCC than in any other liver disease (P < 0.01). Of the 97 patients with HCC, 54 (55.7%) were PIVKA‐II positive, 76.3% had serum concentrations of either PIVKA‐II or α‐fetoprotein, indicating the usefulness of both tumour markers in the diagnosis of HCC.

A simplified method for detecting macro-amylasemia by measuring serum amylase activity at different reaction temperatures

SummaryAmylase activity in serum and urine, and isoamylase, were measured in 300 patients with abdominal pain to detect cases of macroamylasemia. Of these patients, 9 had hyperamylasemia and 2 were diagnosed as cases of macroamylasemia on the basis of their amylase/creatinine clearance ratio, the gel filtration pattern of their amylase on a dextran column, and results of immunological analysis.Amylase activity in macroamylasemia is reported to show an anomalous response to increase in reactiontemperature. In this report, measurements of the temperature-activity relationships of serum amylase confirmed that the ratio of serum amylase activity at 50°C to that at 25°C (AMY-50°C/AMY-25°C ratio) in patients with macroamylasemia was higher than that in normal subjects or patients with pancreatitis. Moreover, when macromolecular amylase in the sera of patients with macroamylasemia was separated from amylase of normal molecular weight by dextran gel chromatography, it showed a significantly higher AMY-50°C/AMY-25°C ratio than the latter. Measurement of this AMY-50°C/AMY-25° ratio seems to be a convenient and useful method for differential diagnosis of hyperamylasemia.