Shin-ichiro Sato

Primary hepatic lymphoma associated with primary biliary cirrhosis

We report a case of primary hepatic lymphoma in a 55-yr-old female patient with primary biliary cirrhosis and Sjögrens syndrome. On July 1994, a tumor measuring 11 mm in diameter was detected in the right lobe of the liver by abdominal ultrasonography. A needle biopsy specimen showed the lesion to contain small- and medium-sized lymphoid cells without obvious atypia, and a provisional diagnosis of pseudolymphoma was made. About 2 yr later, the tumor increased to 15 mm in diameter, necessitating a second needle biopsy. Histological and genetic examinations confirmed non-Hodgkins lymphoma of diffuse, mixed small and large cell, B-cell type. However, the size of the tumor remained almost stable (16 mm in diameter) over a period of 7 months after diagnosis, without any treatment for lymphoma, indicating a low grade malignancy. We document hepatic lymphoma as an additional complication of primary biliary cirrhosis.

Immunoreactivity to monoclonal antibody, Hep Par 1, in human hepatocellular carcinomas according to histopathological grade and histological pattern.

We evaluated the immunoreactivity to monoclonal antibody for hepatocyte (Hep Par 1) and determined the cellular distribution of the antigen in hepatocellular carcinoma (HCC), based on the histopathological grade and histological pattern. The pathological material included 100 areas selected at random in 61 tissue sections from 12 autopsy livers with HCC. Immunoreactivity was classified into four categories; strongly positive (>90% of the area showed positive staining), moderately positive (5-90% of the area), weakly positive (<5% of the area), and negative (completely negative). All 19 (100%) well-differentiated, trabecular type HCC areas were strongly positive for Hep Par 1. Among 11 well-differentiated, pseudoglandular type HCC areas, 2 (18%) were strongly positive, 5 (46%) were moderately positive, 2 (18%) were weakly positive, and 2 (18%) were negative. Among 36 moderately differentiated, trabecular type HCC areas, 6 (17%) were strongly positive, 17 (47%) were moderately positive, 9 (25%) were weakly positive, and 4 (11%) were negative. None of the four moderately differentiated, pseudoglandular type HCC areas were strongly positive, 3 (75%) were moderately positive, 0 was weakly positive, and 1 (25%) was negative. Among 25 poorly differentiated, compact or trabecular type HCC areas, 15 (60%) were weakly positive and 10 (40%) were negative. All 5 (100%) undifferentiated HCC areas were negative for Hep Par 1. Our results indicate that immunoreactivity to Hep Par 1 in HCC decreases with reduced differentiation of the tumor, suggesting that Hep Par 1 monoclonal antibody is useful as a marker for the diagnosis and differentiation of HCCs.

A novel prostaglandin E receptor subtype agonist, 0N0-4819, attenuates acute experimental liver injury in rats

We evaluated the efficacy of ONO-4819, a newly developed agonist of a prostaglandin receptor subtype (EP4), on experimental model of acute liver injury in rats. Acute liver injury was induced by simultaneous intraperitoneal (i.p.) administration of D-galactosamine (GalN, 1 g/kg body weight) and lipopolysaccharide (LPS, 100 mg/kg body weight). The rats received a single intraperitoneal injection of ONO-4819 (0.2 mg/kg body weight) or physiological saline immediately after GalN/LPS administration. Submassive hepatic necrosis with marked elevation of serum total bilirubin, serum aspartate aminotransferase and serum alanine aminotransferase levels developed 24 h after GalN/LPS administration. The administration of ONO-4819 significantly inhibited the development of submassive hepatic necrosis and inhibited the elevation in levels of biochemical markers that indicate liver function. In addition, the apoptotic index of hepatocytes assessed by the TUNEL method was significantly lower in rats treated with ONO-4819 than in the control. Although serum levels of tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma) and interleukin-8 (IL-8) were markedly elevated after GalN/LPS administration, ONO-4819 significantly inhibited the elevation of those of TNF-alpha and IFN-gamma but not that of IL-8. The beneficial effect of ONO-4819 for acute liver injury was similar at doses of 0.1, 0.05 and 0.01 mg/kg body weight. These results suggest that the EP4 agonist, ONO-4819, may have a protective effect against experimental liver injury in rats through the suppression of inflammatory cytokines.

A male patient with severe acute hepatitis who was domestically infected with a genotype H hepatitis B virus in Iwate, Japan

Although all eight genotypes of hepatitis B virus (HBV) strains are circulating in Japan, no cases of acute hepatitis with foreign HBV strains of genotype H have thus far been reported in Japan. Here, we report a 35-year-old Japanese patient with severe acute hepatitis who was domestically infected with genotype H HBV. On admission, he had a high HBV load of 1.0 × 109 copies/ml, elevated levels of total bilirubin (7.0 mg/dl) and alanine aminotransferase (3606 IU/l), and reduced prothrombin activity of 39.0%. The HB-JAIW05 isolate obtained in the present study was composed of 3215 nucleotides and had the highest similarity of 99.7% with the reported genotype H HBV isolate recovered from a Japanese blood donor. The HB-JAIW05 isolate had neither precore (A1896) nor core promoter (T1762/A1764) mutations. However, upon comparison with the consensus sequence of ten reported HBV isolates of the same genotype, the HB-JAIW05 isolate had 17 nucleotide substitutions including five missense mutations in the P gene, which may be related to vigorous replication of HBV in this case. He had no history of traveling abroad, but had had extramarital sexual contact with two Japanese women living in Iwate, Japan, 2 weeks and 2 months before the disease onset, respectively. Our results suggest that rare HBV genotypes such as H may be spreading in Japan via sexual contact. Further molecular epidemiological studies on HBV to clarify the exact changing profiles of de novo HBV infection in Japan in relation to genotype and genomic variability are warranted.

Bile canaliculi-like lumina in fibrolamellar carcinoma of the liver: A light- and electron-microscopic study and three-dimensional examination of serial sections

Flbrolamellar carclnoma (FLC) Is a varlant of hepatocellular carcinoma characterlzed by dlstlnct pathologicel features. The presence of Intracellular lumlna resembling blle canalicull was previously reported in tumor cells of FLC on electron microscopy. Using light microscopy, we describe the presence of intracellular lumina In FLC, which was resected from a 15‐year‐old Japanese girl, as round structures lined with a brush‐like border. These lumina occasionally contained blle. Light microscopic examination of 1 μm thick serial sections of Eponembedded tissue samples showed that the lumina were located in the intracellular space without any connection to the Intercellular space. However, we also detected a small number of lumlna that were lined by microville which were present between adjacent tumor cells. Results suggest that the presence of the Intracellular Iumlna in tumor cells probably represents a common histopathologic feature of FLC.

Beneficial effect of cyclosporin A on acute hepatic injury induced by galactosamine and lipopolysaccharide in rats

Cyclosporin A (CsA), a potent immunosuppressive agent, is used clinically to prevent allograft rejection after organ transplantation. Although recent experimental studies show that CsA prevents severe hepatic injury associated with tumor necrosis factor-alpha (TNF-alpha), the efficacy of CsA has not been confirmed. In the present study, we evaluated whether CsA protects against the severe hepatic injury induced by simultaneous administration of D-galactosamine (GaIN, 200 mg/kg) and lipopolysaccharide (LPS, 10 µg/kg) into the portal vein. The method of CsA (10 mg/kg) was divided into four groups; i.e. preinjection (24 and 2 h before administration of GaIN and LPS), concomitant single-injection, multiple-interval injection (0, 3, 6, 9 and 15 h after administration of GaIN and LPS) and no treatment. In the group receiving multiple-interval injections of CsA, liver function parameters (total bilirubin, asparate aminotransferase and alanine aminotransferase levels in sera) were significantly improved and the development of massive hepatic necrosis was significantly prevented, but no improvement of liver function parameters or histological changes was shown either the pre- or concomitant single-injection groups. The serum levels of TNF-alpha and interferon-gamma showed no differences between the no treatment and multiple-interval injection groups. However, the serum level of interleukin 8 and neutrophil infiltration into the liver tissue after 24 h GaIN and LPS administration were significantly lower than those of the control group. The apoptotic index of liver tissue using the TUNEL method was not significantly different. Our data suggest that CsA protects against acute hepatic injury, if it is administered at the appropriate time during the course of hepatic injury.

Distinguishing small lymph vessels in the portal tracts of human liver from portal veins by immunohistochemistry for α smooth muscle actin

Abstract Differentiation between hepatic lymph vessels and portal vein branches is difficult to determine by light microscopy. We tested the usefulness of immunohistochemical staining for a smooth muscle actin of human liver tissue in differentiating lymph vessels from small portal vein branches. A gelatin solution containing barium sulfate as a marker was infused into the lymph vessels on the surface of autopsy livers or into portal vein branches. Immunohistochemical staining for α smooth muscle actin was performed on formalin-fixed, paraffin-embedded tissue sections. We evaluated the reaction in walls of vessels identified as lymph vessels or portal vein branches by using barium-gelatin. A total of 174 lymph vessels and 165 portal vein branches was evaluated in three autopsy livers. The walls of the lymph vessels, measuring from 6.5–92.3 μm in diameter, were negative for a smooth muscle actin in all specimens, while those of portal vein branches, measuring from 15.1–698.9 μm, appeared circumscribed or more than half positive. Using immunohistochemistry for α smooth muscle actin, lymph vessels less than 100 μm in diameter can be distinguished from portal vein branches in the portal tracts of human liver by the presence or absence of reaction to a smooth muscle actin.

Immunohistochemical study of tumor necrosis factor-alpha in acute liver injury induced by Propionibacterium acnes and lipopolysaccharide in rats

Abstract The effect of intravenous injection of Propionibacterium acnes (P. acnes) and lipopolysaccharide (LPS) on the distribution of tumor necrosis factor-α (TNF-α) in different organs have not previously been investigated. Immunohistochemistry and histological examination were employed in evaluating the distribution of TNF-α in the liver, spleen, lungs and bone marrow in rats injected intravenously with P. acnes followed by LPS 7 days later. Granulomas containing ED1-positive macrophages were observed in the liver 7 days after P. acnes injection. Subsequent LPS injection resulted in proliferation of ED1-positive macrophages in the sinusoids and coagulation necrosis of hepatocytes after 6 h. TNF-α was detected in ED2-positive macrophages (Kupffer cells) 1 day after P. acnes injection and in macrophages constituting the granulomas 7 days later, but prior to LPS injection. TNF-α was also detected in ED1-positive macrophages in the spleen, predominantly in the marginal zone. When granulomas were formed 7 days after P. acnes injection, TNF-α was observed in macrophages of the granulomas. TNF-α was also detected in macrophages of the granulomas found in the lung 1 day after P. acnes injection. No macrophages expressing TNF-α were found in the granulomas of bone marrow. The highest expression was in the liver at any time interval and in macrophages constituting granulomas. Our results suggest that the high expression of TNF-α in the liver results in selective hepatic necrosis. The expression of TNF-α in macrophages of the liver after P. acnes injection and the subsequent development of hepatic necrosis after LPS injection suggest that P. acnes acts as an inducer of TNF-α production in macrophages while LPS acts as a trigger for the release of TNF-α from macrophages.

Endoscope Disinfection with Acid Electrolyzed Water

Reports of endoscope‐mediated infectious diseases have been increasing. In order to investigate the usefulness of acid electrolyzed water for endoscope disinfection, a test endoscope was disinfected with acid electrolyzed water, and bacteria were examined before and after disinfection. The rates of detection of general bacteria on the surface of the endoscope were 100% (33/33 cases) immediately after its removal, 73% (24/33 cases) after washing the endoscope, and 0% (0/33 cases) after disinfection with acid electrolyzed water. The rates of detection for the forceps channel were 100% (33/33 cases) immediately after endoscope removal, 100% (33/33 cases) after washing, and 0% (0/33 cases) after disinfection with acid electrolyzed water. Helicobacter pylori (H. pylori) was detected at the tip of the endoscope in 3 of the 33 cases immediately after the endoscope was removed. The bacteria were observed in 3 cases after washing as well, but disappeared after disinfection with acid electrolyzed water. The bactericidal effect of acid electrolyzed water on H. pylori was also investigated by culturing a condensed solution of a standard strain of H. pylori with acid electrolyzed water. Growth was inhibited within 30 seconds after the addition of acid electrolyzed water. Endoscope disinfection with acid electrolyzed water should be considered useful.