Shinichi Tozuka

Mutations in the core promoter region of hepatitis B virus in patients with chronic hepatitis B

The core promoter region of hepatitis B virus genomes regulates transcription of the precore and pregenomic mRNAs encoding hepatitis B e antigen (HBeAg) and core antigen that contain target epitopes for cytotoxic T lymphocytes. The prevalence and clinical significance of mutations in this region were investigated. DNA was extracted from six asymptomatic carriers positive for HBeAg, eight asymptomatic carriers positive for an anti‐HBe antibody, and 24 patients with chronic liver disease. The core promoter and precore regions of hepatitis B virus genomes were amplified by polymerase chain reaction, and predominant sequences were determined by direct sequencing. Mutations were found in none of the HBeAg‐positive asymptomatic carriers but in all of the anti‐HBe‐positive asymptomatic carriers and the patients with chronic liver disease. Especially, A to T mutations at nucleotide 1762 and G to A mutations at nucleotide 1764 were found in five anti‐HBe‐positive asymptomatic carriers, and 22 patients with chronic liver disease. These two mutation hot spots were located within binding sites of the nuclear factors, and nucleotide 1762 was also involved in the A, T rich sequence that is located 28 base pairs upstream of the precore mRNA initiation site. Serum HBeAg and DNA polymerase levels were significantly lower in patients with these mutations than those without these mutations, and five individuals with these mutations were positive for anti‐HBe despite the absence of the precore stop codon mutation. These mutants may be selected by host immune response to HBeAg and/or core antigen.

Effects of additional administration of colchicine in ursodeoxycholic acid-treated patients with primary biliary cirrhosis: a prospective randomized study

BACKGROUND/AIMS Although ursodeoxycholic acid is effective for the treatment of primary biliary cirrhosis, some patients do not respond to this treatment. The objective of the present study was to investigate the effects of additional administration of colchicine in ursodeoxycholic acid-treated patients with primary biliary cirrhosis. METHODS Twenty-two patients with primary biliary cirrhosis treated with ursodeoxycholic acid (600 mg/day) for 30 months were randomly assigned to two groups: group 1, colchicine (1 mg/day) and ursodeoxycholic acid (n = 10); group 2, ursodeoxycholic acid alone (n = 12). RESULTS In group 1, there were significant decreases in mean serum levels of alkaline phosphatase, total bilirubin, gamma-glutamyltranspeptidase, alanine aminotransferase, aspartate aminotransferase, and IgM, and these changes were more remarkable in those who responded poorly to ursodeoxycholic acid. In contrast, there were no significant changes in those values in group 2. CONCLUSIONS Additional administration of colchicine to ursodeoxycholic acid may be beneficial for patients with primary biliary cirrhosis, especially those who respond poorly to ursodeoxycholic acid. It is necessary, however, to further confirm the efficacy of colchicine by examining histological changes and following the patients for longer periods.

Prostaglandin E-producing hepatocellular carcinoma with hypercalcemia

An autopsy case of prostaglandin E‐producing hepatocellular carcinoma with hypercalcemia is presented in this article. A 72‐year‐old man showed high serum calcium levels (14.2 to 17.3 mg/100 ml) and hypophosphatemia. The plasma level of immunoreactive parathyroid hormone was below the normal range. Administration of oral indomethacin 50 mg daily was effective in decreasing the serum calcium concentration. However, this effect lasted only 5 days, after which it returned to pretreatment levels. The patient died in a hypercalcemic coma. By an autopsy, hepatocellular carcinoma was found in the right lobe of the liver. However, no obvious bone metastases nor abnormalities in the parathyroid glands were detected. The immunoreactive prostaglandin E level assayed in the neoplastic tissue (2278 ng/g) was significantly high when compared with level in the nonneoplastic liver tissue (194 ng/g). The production of prostaglandin E by the tumor itself appears to be the most likely mechanism for the hypercalcemia in this patient.

Severe chronic active hepatitis induced by UFTR containing tegafur and uracil.

SummaryA 77-year-old female patient developed severe hepatic injury after the administration of UFTR, which contains tegafur and uracil, for postoperative chemotherapy of colon cancer. Liver damage was recognized 10 months after its administration. Serum markers for viral hepatitis and various autoantibodies were negative. The wedged biopsied liver specimen revealed advanced chronic active hepatitis with periportal confluent necrosis, marked intralobular spotty necrosis, and significant proliferation of pseudo-bile ductules. Although the cessation of the drug and conservative therapies improved hepatic function, an accidental readministration of UFTR caused her severe hepatic damage again. These findings suggest that liver injury in the present case was caused by UFTR. Histological findings were unique. Although tegafur is known to worsen hepatic function when given to patients with liver cirrhosis, UFTR may also cause severe hepatic injury in those without preexisting liver disease.

Hepatocellular carcinoma producing carcinoembryonic antigen

SummaryA 78-year-old female with hepatocellular carcinoma and high serum levels of carcinoembryonic antigen is reported. Using a immunohistochemical technique, CEA was demonstrated within the cytoplasm of the tumor cells.

Improvement of peripheral blood lymphocyte subsets in primary biliary cirrhosis after ursodeoxycholic acid therapy

Immunological abnormalities frequently observed in patients with primary biliary cirrhosis are considered to be related to the pathogenesis of this disease. We performed a prospective trial to evaluate whether immune mechanisms play a role in the effectiveness of ursodeoxycholic acid (UDCA) therapy. Fifteen female patients with primary biliary cirrhosis were followed for 1 year and were then treated with UDCA (600 mg/day) for another year. Laboratory tests, including peripheral blood lymphocyte subsets assessed by dual colour fluorescence analysis using monoclonal antibodies against respective T cell markers, were evaluated at the beginning of the study, at the start of therapy and at the end of therapy. In primary biliary cirrhosis, the proportion of cytotoxic T cells, suppressor inducer T cells and αβ‐receptor bearing T cells were significantly lower than in healthy controls. No significant changes were observed in the proportions during the year before the therapy. These reductions, however, recovered to normal ranges after 1 year of UDCA therapy. These changes were associated with an improvement in the serum levels of aspartate aminotransferase, alkaline phosphatase, γ‐globulin and IgM. The close correlation between the improvement in the imbalance of lymphocyte subsets after the therapy and the clinical status suggests that an immunological process may play a role in the effectiveness of therapy in primary biliary cirrhosis.

Case Report: A Variant Alkaline Phosphatase-Producing Gastric Carcinoma with Super Bone Scan

in English) 4. Uchida T, Shikata T, Shimizu S, Takimoto Y, Iino S, Suzuki H, Oda T, Hirano K, Sugiura M: Gonadotropin and alkaline phosphatase producing occult gastric carcinoma with widespread metastasis of generalized bone. Cancer 48:140 ± 150,

Flow Cytometric Method to Detect Lymphocyte Transformation in Drug-Allergic Hepatic Injury

Flow cytometric methods for the analysis ofincorporated bromodeoxyuridine are extremely rapid andsimple. We investigated whether these methods wereuseful for detecting drugallergic hepatic injury in 18 patients with drug-allergic hepatic injury,18 healthy controls, and 9 nonallergic patientsreceiving drugs. Peripheral blood mononuclear cells werestimulated with drug solutions. Incorporation ofbromodeoxyuridine was detected after labeling with FITC, andS-phase cells were counted by flow cytometry.Percentages of S-phase cells in drug-stimulated cultureminus those in spontaneous cultures were less than 1% in both healthy controls and nonallergic patientsreceiving drugs. Taking 1% as the upper limit, 13patients (72%) were judged as positive. After the invitro addition of interleukin-2, two patients among five who had been judged as negative werejudged as positive. Lymphocyte transformation test byflow cytometry may be useful in the diagnosis ofdrug-allergic hepatic injury.

Case Report: Gastric Adenocarcinoma Producing Neuron-Specific Enolase

Enolase s (2-phospho-D-glyce rate hydro-lyase , EC 4.2.1.11) are glycolytic enzymes that convert 2-phosphoglyc erate to phosphoenolpyruvate . They are dimers composed of three immunological ly distinct subunits a , b , and g (1, 2). The g g and a g -enolase s are expressed preferentially in neurons and neuroendocrine cells (3, 4), and neuron-spe ci® c enolase (NSE) has often been used to refer to the g g -enolase or g -subunit itse lf (5). Tumors with neuroendocrine characte ristics, including small cell carcinoma of the lung and neuroblastoma contain high concentrations of NSE (6, 7). A majority of patients bearing such tumors have high serum NSE leve ls (8 ± 10) . Although neuroendocrine granule s were often demonstrable ultrastructural ly in gastric endocrine tumors (11± 14) and a part of a solid carcinomao of the stomach (15) , gastric adenocarcinoma with high serum NSE leve ls has not been reported. Described in this article is an autopsy case of gastric adenocarcinoma with high serum NSE leve ls. NSE was detected in the carcinoma cells by immunohistoche mistry, which was conside red to result in elevated serum NSE leve ls.