Yoshitane Kosaka

Fulminant hepatitis B: Induction by hepatitis B virus mutants defective in the precore region and incapable of encoding E antigen

Clones of hepatitis B virus were propagated from 10 cases of fulminant hepatitis B after amplification by polymerase chain reaction and their nucleotide sequences of the precore region were determined. All 113 clones from 9 cases had a point mutation from guanine to adenine at nucleotide 83 in the precore region, which converted codon 28 for tryptophan (TGG) to a stop codon (TAG) and prohibited the synthesis and secretion of hepatitis B e antigen. Precore-region defects were not detected in any of 23 clones from the remaining 1 case. By contrast, precore-region defects were not found in any of 180 clones from 8 cases of acute hepatitis B without hepatic failure serving as controls. The source of infection was traceable in 3 cases. The same precore-region defect, along with the sequence identity of 435 nucleotides, was observed in clones from the case of a baby and his grandmother, who carried the virus and was implicated in the transmission, and also in clones from two pediatricians and the carrier patients they attended. These findings support the hypothesis that precore-defective mutants have stronger activity to induce fulminant hepatitis than nondefective viruses.

Infection with GB virus C (GBV-C) in patients with fulminant hepatitis

BACKGROUND/METHODS There appear to be hepatitis viruses other than hepatitis A, B, C, D and E. One of these has been proposed with a designation of GB virus C. Sera from 44 patients with fulminant hepatitis were tested for RNA of GB virus C by reverse-transcription polymerase chain reaction with nested primers deduced from the putative non-structural 3 (helicase) region. RESULTS RNA of GB virus C was detected in three (20%) of 15 patients with hepatitis B virus infection and three (12%) of 25 patients without markers of hepatitis A-E virus infection. Overall, GB virus C RNA was detected in six (14%) of the 44 patients with fulminant hepatitis, at a frequency significantly higher (p < 0.001) than that in three (0.9%) of 326 blood donors matched for age with the patients. CONCLUSIONS These results indicate a role of GB virus C in inducing fulminant hepatitis either by itself or in concert with the other hepatitis viruses.

Evaluation of des-γ-carboxy prothrombin as a marker protein of hepatocellular carcinoma

We measured des‐γ‐carboxyglutamic acid prothrombin (protein induced by vitamin K absence or antagonist‐Factor II: [PIVKA‐II]) in plasmas of normal subjects, patients with thrombotic disease, those with hepatic disease including hepatocellular carcinoma, and those with carcinoma of other tissues, and compared the results with results of blood coagulation tests used for the examination of hepatic function. In addition, in the patients with hepatic disease, PIVKA‐II and α‐fetoprotein (AFP) levels were compared. The PIVKA‐II level was frequently high in patients with thrombotic disease given warfarin therapy and those with hepatocellular carcinoma. However, in patients with thrombotic disease who were not given warfarin therapy, no significant correlation was seen between the PIVKA‐II value and the results of the thrombotest or hepaplastin test, suggesting no association between the PIVKA‐II level and the degree of impairment of hepatic function. In 70 patients with hepatocellular carcinoma, the percentage of patients positive for PIVKA‐II (⩾0.1 μg/ml) and those positive for AFP (⩾20 ng/ml) were similar (77% and 74%, respectively). Pearsons correlation of coefficient between the PIVKA‐II value and the AFP value in the 70 patients was 0.463. However, false‐positive rates in patients with hepatic disease other than hepatocellular carcinoma were lower for PIVKA‐II. Combined assessment of PIVKA‐II and AFP increased positive rates and allowed exclusion of false‐positive patients. The plasma PIVKA‐II level is suggested to be useful as an indicator of warfarin control in patients with thrombotic disease, as a marker of hepatocellular carcinoma, and is particularly of value when assessed in combination with AFP. Cancer 64:2546–2551, 1989.

Herbal medicine ‘Sho‐saiko‐to’ induces tumour necrosis factor‐α and granulocyte colony‐stimulating factor in vitro in peripheral blood mononuclear cells of patients with hepatocellular carcinoma

‘Sho‐saiko‐to’ (TJ‐9) is a Japanese herbal medicine that is commonly administered to patients with chronic viral liver disease in order to improve their overall physical condition and to prevent the development of liver cancer. The present in vitro study demonstrated that, by adding TJ‐9 to cell cultures, there were dose‐dependent increases in production levels of tumour necrosis factor‐α (TNF‐α) and granulocyte colony‐stimulating factor (G‐CSF) in peripheral mononuclear cells of patients with hepatocellular carcinoma accompanied by liver cirrhosis. Increases in the production of TNF‐α and G‐CSF in control cell cultures exposed to different herbal medicines were low, and this indicates the specificity of the responce increases in production of these cytokines to TJ‐9. TNF‐α and G‐CSF are known to play important roles in the biological defence mechanism. Administration of TJ‐9 may, therefore, be beneficial for patients afflicted with intractable liver diseases because it could mildly induce these cytokines.

Analysis of serum cytokine levels in primary biliary cirrhosis patients and healthy adults

By using commercially available ELISA kits, serum IL‐6 and TNF‐α levels in healthy adults, and the levels of various cytokines in patients with primary biliary cirrhosis or chronic viral liver diseases, were investigated. IL‐6 levels of healthy subjects were distributed in a wide range, and the distribution pattern was similar to those of the patients. TNF‐α levels tended to be low in females in their 30s, but there were no abnormalities in the patients. Characteristic findings, in the primary biliary cirrhosis patients, were an increase of IFN‐γ and IL‐2 levels, and a decrease of GM‐CSF levels (P<0.05). IL‐8 levels were higher in the patients than in the healthy subjects (P<0.05), and the increase was remarkable in chronic viral liver disease patients. We believe that measurement of serum cytokine levels as a clinical immunological test is highly useful. Further development of simpler, more rapid, and more sensitive analysis methods is desired. J. Clin. Lab. Anal. 12:77–82, 1998.

Prognostic value of peritoneoscopic findings in cirrhosis of the liver

The prognostic value of peritoneoscopy was examined in 372 patients with liver cirrhosis according to the degree of development and size of regenerating nodules, the development of reddish markings, the development of small lymphatic vesicles, the presence or absence of patchy markings, the size of the right and left hepatic lobes, and the degree of splenomegaly. The cumulative survival rate was compared with these peritoneoscopic parameters. The usefulness of peritoneoscopic and histological findings in the prognosis of liver cirrhosis was evaluated using the proportional hazard model of Cox. Significant differences were observed in the cumulative survival rate with respect to the degree of development of regenerating nodules, the size of the right hepatic lobe, the formation of small lymphatic vesicles, and the degree of splenomegaly. Analysis using Coxs proportional hazard model indicated that peritoneoscopic findings are of greater clinical use than histological findings in determining the prognosis of liver cirrhosis.

Therapeutic Effects of Tiopronin on Chronic Hepatitis: A Double-Blind Clinical Study

The efficacy of tiopronin (2-mercaptopropionyl glycine) 600 mg daily in the treatment of chronic active or chronic persistent hepatitis has been assessed in a double-blind controlled clinical trial of 12 weeks involving 165 Japanese patients with histologically proven disease. Treatment with the drug was associated with a significant improvement in abnormalities of serum transaminase and gamma glutamyl transpeptidase, with reversion towards baseline values on stopping the drug. Improvement was independent of the histological classification of the disease, or HBsAg status. The drug was well tolerated with few side-effects. The results of this short-term study indicate that tiopronin may be of value in the treatment of chronic hepatitis.

Fulminant hepatic failure caused by ecarazine hydrochloride (a hydralazine derivative)

The cause of fulminant hepatic failure is reported to be unknown in more than half the cases in Japan. We recently reviewed 23 cases of fulminant hepatic failure that had been treated at our hospital. The cause of disease had been regarded as unknown before this study. It was found that seven of these patients had been under ecarazine hydrochloride therapy when they developed fulminant hepatic failure. We examined the reasons why fulminant hepatic failure in these seven patients had not been previously attributed to ecarazine, and found that it could be explained by the following factors: (1) the time from the start of ecarazine therapy to the onset of hepatic failure was long; (2) in all cases, hepatic failure developed more than 10 days after the clinical recognition of hepatitis; and (3) characteristic signs of drug‐induced hepatic failure such as a skin rash and positive lymphocytes stimulation test with the drug were absent in all cases. Fulminant hepatic failure in these cases could be characterized by: (1) rapid decrease in serum alanine transaminase (ALT) level after discontinuation of ecarazine, (2) prolonged jaundice despite discontinuation of ecarazine, (3) high incidence of anti‐nuclear antibody (ANA) (57%), and (4) histological findings of extensive hepatocellular necrosis ranging from bridging necrosis to massive necrosis. Of the seven patients, four died of fulminant hepatic failure. These four patients had received high doses of ecarazine hydrochloride for prolonged periods. Our data suggest that there may be many cases in which the cause of fulminant hepatic failure or acute hepatitis was not previously determined that can be attributed to long‐ term drug therapy for chronic diseases.

Effects of the Japanese herbal medicine ‘Sho-saiko-to’ as a cytokine inducer

The herbal medicine, Sho-saiko-to (TJ-9), has been widely prescribed to chronic viral liver disease patients in Japan. This study examined the inductions of such sytokines as interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and granulocyte-colony stimulating factor (G-CSF), on some fractions of peripheral blood mononuclear cells (PBMC) by TJ-9 and each of its seven components. IL-1β, TNF-α, and G-CSF were highly induced by scutellaria root and glycyrrhiza root on monocytes/macrophages. By repeating the same experiments using taxol (an LPS antagonist)-treated substances, authors confirmed that these inductions were not attributable to the presence of quite low LPS in TJ-9 solution, and the cytokine inductions are the specific effect of TJ-9. Because TJ-9s macrobiotic effect in liver cirrhosis patients has been proven statistically in an etiological study, TJ-9 could be a new important therapy in chronic liver diseases.

An effective intradermal hepatitis B vaccination

Small dose intradermal (i.d.) inoculation methods of hepatitis B vaccine have been reported to be effective and economical. We determined the best method to obtain high antibody levels within a short period of time and for the long-term maintenance of these levels. A total of 173 female students were randomly allocated to seven groups: six i.d. inoculation groups, to which 6-12 micrograms was administered in three or four divided doses (Groups A-F), and a control group (Group G) which received three 10 micrograms intramuscular (i.m.) inoculations. Serum hepatitis B antibody levels were quantified in weeks 4 and 8, and months 4, 7, and 12. Positivities in all groups were not significantly different at each measurement time. In month 4, geometric mean antibody levels in the three i.d. groups (10-12 micrograms in three divided doses; 79.1-107.0 IU l-1) were significantly higher than in Group G, which had received two of three i.m. injections (17.6 IU l-1; P < 0.01, P < 0.05). In the group which received four 2 micrograms i.d. inoculations, the level was higher than in Group G in month 7, but lower in month 12. It was concluded that three i.d. inoculations, each of 4 micrograms, may be used to obtain high antibody levels within a short period of time. However, it is recommended that a 10 micrograms i.m. injection in month 6 is applied as a booster. Consequently, we could not present an economic and effective low-dose intradermal inoculation method.