Relation between markers for viral hepatitis and clinical features of Japanese patients with hepatocellular carcinoma: possible role of alcohol in promoting carcinogenesis.

To assess the relationship between hepatitis virus markers and the clinical features of hepatocellular carcinoma (HCC), we measured markers for hepatitis B virus (HBV) and hepatitis C virus (HCV) in 88 Japanese patients with HCC. Twelve (14%) patients were HBsAg-positive and 67 (76%) were anti-HCV-positive (both c100-3 and c11/c7). HCV-RNA was detected in 8 (38%) of the 21 anti-HCV-negative patients by PCR, so that 75 patients (85%) were infected with HCV. Of the HBsAg-negative patients infected with HCV with no history of blood transfusion, the mean age of the alcoholics (consumption > 80 g ethanol daily for at least 10 years) was lower than that of the non-alcoholics (60 years vs. 65 years, P < 0.05). Among the HBsAg-negative and anti-HCV (or HCV-RNA)-positive patients with a history of blood transfusion, the mean interval between the time of blood transfusion and the diagnosis of HCC in the alcoholics was shorter (21 years) than that in the nonalcoholics (27 years), but the difference was not statistically significant. We conclude that infection by both HCV and HBV may play a role in the development of HCC, and that alcohol consumption may promote carcinogenesis.

Aspartate aminotransferase-linked immunoglobulin complexes in serum of a patient with primary biliary cirrhosis.

A 51-year-old woman who had been treated for primary biliary cirrhosis (PBC) was admitted to our hospital for evaluation of unexplained, isolated, persistently increased aspartate aminotransferase (AST) activity. Results of laboratory tests on admission showed: AST 171 KU, alanine aminotransferase 28 KU, and anti-mitochondrial titer 1/1280. Results of hepatitis B surface antigen (HBs Ag) and hepatitis C virus antibody (HCV Ab; C100-3) assays were negative. Histology of a liver biopsy specimen was compatible with a diagnosis of PBC (stage III of Scheuer's classification). The molecular size of serum AST was estimated to be more than 500,000 by high-performance size-exclusion liquid chromatography. Electrophoretic analysis showed an abnormal band of AST between supernatant AST (sAST) and mitochondrial AST (mAST), which band was characteristic of AST-immunoglobulin complexes (AST-Ig). Ouchterlony double-diffusion and immunoprecipitation tests identified the immunoglobulin component as IgM. The presence of AST-Ig appeared to be responsible for the elevated serum AST.

Development of pseudolymphoma of liver following interferon-alpha therapy for chronic hepatitis B.

A 42-year-old woman with biopsy-proven chronic hepatitis B, who had been treated with human leukocyte-derived interferon-alpha (huLe-IFN alpha) therapy for two months was found to have liver tumors on routine abdominal ultrasonography examination. She underwent laparotomy, and partial hepatectomy was performed under the clinical diagnosis of hepatocellular carcinoma. The lesions were diagnosed histologically as pseudolymphoma based on the massive infiltration of small mature lymphocytes and the presence of hyperplastic lymph follicles with germinal centers. Immunohistochemistry revealed polyclonal origin of the involved lymphocytes. The possible association between IFN alpha treatment and chronic hepatitis B with the development of pseudolymphoma is discussed.

Discrimination of liver cirrhosis from chronic hepatitis by analysis of serum cholinesterase isozymes using affinity electrophoresis with concanavalin A or wheat germ agglutinin.

Isozymic alteration of serum cholinesterase (ChE) was investigated in patients with chronic liver diseases using affinity electrophoresis with concanavalin A (Con A) or wheat germ agglutinin (WGA). On Con A-containing agarose gel electrophoresis, three bands with enzyme activity (named bands I to III, from the anodic side to the cathodic) were observed in sera of normal controls. Disappearance of band II was observed in 50% (15/30) of cirrhotic patients, but only one of 20 patients with chronic hepatitis lacked band II of the serum ChE isozymes. Meanwhile, WGA-containing agarose gel electrophoresis revealed that normal controls had four ChE isozymes (named bands I to IV from the anodic side to the cathodic). These four isozymes were also observed in patients with chronic hepatitis. However approximately 67% (20/30) of cirrhotic patients lacked band II of ChE isozymes. When these two affinity electrophoreses were used in combination, 22 (73%) of 30 cirrhotic patients had isozymic alteration of their serum ChE on either Con A-containing or WGA-containing agarose gel electrophoresis, or both. Thus, affinity electrophoreses with Con A and WGA seemed to be useful methods in differentiating liver cirrhosis from chronic hepatitis.

[Statistical analyses of clinical observations made on outpatients visiting the Department of Oral Diagnosis, Meikai University School of Dentistry Hospital, for the past 16 years].

Statistical analyses of clinical observations made on outpatients visiting the Department of Oral Diagnosis, Meikai University School of Dentistry Hospital from June, 1970, to August 1986 were carried out. The total number of new outpatients during this period was 73,708. The results were as follows: 1) The largest number of patients was found in 1978. 2) There was a tendency for the number of new patients to increase in March, June, and August of every year. The smallest number of patients came in December. 3) The greatest number of new patients visiting the hospital was in the 0-9 age category, followed by those in the 20-29 and 30-39 age categories. 4) The greatest chief complaint of patients visiting the hospital was pain, followed by masticatory disturbance and swelling. 5) The number of outpatients coming to the hospital for cosmetic and temporomandibular joint problems has been increasing on a yearly basis over the past 16 years.

Serum pseudouridine as a biochemical marker in patients with hepatocellular carcinoma.

The concentration of serum pseudouridine, a degradation product of transfer ribonucleic acid, was determined by high-performance liquid chromatography in patients with hepatocellular carcinoma, liver cirrhosis, other benign hepatobiliary diseases, and healthy controls. The serum pseudouridine concentration in patients with hepatocellular carcinoma was significantly higher than that in patients with cirrhosis or the controls. Twenty-seven (51.9%) of 52 patients with hepatocellular carcinoma had serum pseudouridine concentrations higher than the mean value for healthy controls plus 2 SD. Fourteen of the 36 patients who had serum alpha-fetoprotein levels below 400 ng/ml, had elevated serum pseudouridine concentration. In total, 36 of the 52 patients (69.2%) could be detected by combination of these two markers. Two patients who had developed hepatocellular carcinoma during the course of cirrhosis and were continuously negative for alpha-fetoprotein, had higher levels of the pseudouridine concentration when hepatocellular carcinoma occurred. Furthermore, 4 of the 7 patients who had a very small cancer and were negative for alpha-fetoprotein, had elevated serum pseudouridine concentration. These results indicate that serum pseudouridine is a useful biochemical marker and that serum pseudouridine and alpha-fetoprotein in combination are considered to serve as complementary markers, for the diagnosis of hepatocellular carcinoma.

Isocholic acid formation from 7 alpha,12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid with human liver enzyme.

The formation of isocholic acid from 7 alpha, 12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid by human liver preparations was examined in vitro. Liver preparations were incubated with 7 alpha, 12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid at pH 7.4 in a phosphate buffer containing NADPH or NADH. The products formed were analyzed by gas chromatography and gas chromatography/mass spectrometry. Results showed that 7 alpha,12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid was reduced mainly to isocholic acid and to cholic acid in a smaller amount in the presence of NADPH, while it was reduced only to cholic acid in the presence of NADH. The reducing enzyme participating in the formation of isocholic acid was localized largely in the cytosol and had more specificity to the unconjugated form as substrate than to the conjugated forms. 3-Keto bile acid analogues, 3-keto-5 beta-cholanoic and 7 alpha-hydroxy-3-keto-5 beta-cholanoic acids were not reduced to the corresponding iso-bile acids by the cytosol in the same conditions used in the isocholic acid formation and the activity of the enzyme catalyzing the reduction of 7 alpha,12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid to isocholic acid was not inhibited by the addition of 3-keto-5 beta-cholanoic acid or 7 alpha-hydroxy-3-keto-5 beta-cholanoic acid to the reaction mixture. Furthermore, on column chromatography of Affi-Gel Blue, the peak of the enzyme catalyzing the reduction of 7 alpha,12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid to isocholic acid was clearly distinguished from that of the enzyme catalyzing the reduction of 3-keto-5 beta-cholanoic acid to isolithocholic acid and that of alcohol dehydrogenase. These results indicate that this enzyme catalyzing the reduction of 7 alpha,12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid to isocholic acid is different from the enzyme(s) catalyzing the reduction 3-keto-5 beta-cholanoic and 7 alpha-hydroxy-3-keto-5 beta-cholanoic acids to the corresponding iso-bile acids and from alcohol dehydrogenase, and has a stereospecific character for 7 alpha,12 alpha-dihydroxy-3-keto-5 beta-cholanoic acid.

[The effect of lithium carbonate against leukopenia during systemic chemotherapy in patients with small cell carcinoma of the lung].

To investigate whether lithium carbonate ameliorates the leukopenia and infectious complication that accompany systemic chemotherapy, we studied 19 patients with small cell carcinoma of the lung receiving combination chemotherapy. Eight patients received systemic chemotherapy and lithium carbonate and 11 patients received systemic chemotherapy alone. The mean leukocyte count nadir during chemotherapy was significantly higher in the patients of the lithium group than in the patients of the control group (p less than 0.05). Percentage of infectious complication related to leukopenia was lower in the lithium group than in the control group, although there was no significant difference between these two groups. There was almost no significant side effect except for liver dysfunction in one patient. We therefore believe that lithium carbonate is an effective and safe drug against leukopenia during cytotoxic chemotherapy.

Acute and subacute toxicity of 2,6-dimethyl-3,5-dimethoxycarbonyl-4-(o-difluoromethoxyphenyl)-1 ,4-dihydropyridine (PP-1466).

The acute and subacute oral toxicity of 2,6-dimethyl-3,5-dimethoxycarbonyl-4-(o-difluoromethoxyphenyl)-1,4 -dihydropyridine (PP-1466) was investigated in several animal species in comparison with nifedipine and nicardipine. A clear species difference in LD50 values was found in acute toxicity of PP-1466, and rabbits were the most sensitive between animal species used, then dogs, mice and rats in order. Prominent acute circulatory failure and associated secondary changes were noticed in toxic signs and autopsy findings. PP-1466 as well as nifedipine was apparently less toxic than nicardipine. In the subacute toxicity studies in rats, deaths occurred only in the 2000 mg/kg/d treated groups of both sexes of PP-1466 and nifedipine. Major changes in various observations and examinations were focussed on the cardiovascular system and liver. On the cardiovascular system, it was revealed as congestion and hemorrhage in the various organs and tissues on autopsy finding in dead rats during the test period. A dose-dependent increase in heart weight was observed in rats sacrificed at the termination of the test period. On the liver, it was revealed as a dose-dependent increase in liver weight, changes in liver lipid levels, changes in several serum biochemistry parameters, such as GOT, GPT and ALP (alkaline phosphatase) activities and lipid levels measured at the termination of the test period. These changes were toxicologically mild and functional except the autopsy findings in dead rats. Female rats were slightly more sensitive than males, and PP-1466 was slightly less toxic than nifedipine on subacute oral toxicity in rats.