Gotaro Toda

AN EPIDEMIC OF INFANTILE PAPULAR ACRODERMATITIS (GIANOTTI'S DISEASE) IN JAPAN ASSOCIATED WITH HEPATITIS-B SURFACE ANTIGEN SUBTYPE ayw

An epidemic of infantile papular acrodermatitis (I.P.A.) (Gianottis disease) occurred in Matsuyama City, in south-east Japan in 1974-75. Patients ages ranged from less than one year to eight years. Hepatitis-B surface antigen (HBsAg) was detected by an immune adherence haemagglutination method in the serum samples of 48 of the 54 patients tested. HBsAg subtypes were determined by a haemagglutination-inhibition method. ayw antigens were identified in 42 patients and adr antigens in 3; it was not possible to determine subtypes in the remaining 3 patients because antigen titres were too low. Since subtype ayw and I.P.A. are extremely rare in Japan, the association of the disease with HBsAg subtype ayw is regarded as being most significant.

A multi-center double-blind controlled trial of ursodeoxycholic acid for primary biliary cirrhosis.

SummaryA multi-center double-blind controlled trial of ursodeoxycholic acid (UDCA) for treatment of primary biliary cirrhosis (PBC) was carried out. Twenty two and 23 patients were treated with 600mg/day UDCA and placebo, respectively, for 24 weeks. In UDCA - treated patients, fall of serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and gamma glutamyltranspeptidase activities started within 4 weeks after start of the trial and continued throughout the trial period. The serum IgM level fell in 7 UDCA-treated patients examined but not in 10 placebo-treated patients examined. Serum bilirubin concentration showed no significant change at the end of the study in either of UDCA- and placebotreated group of patients. There was no significant difference between these two groups with respect to the frequency of improvement of pruritus. In UDCA-treated patients, serum bile acid composition changed markedly, though its concentation showed no significant change. The percentage of total bile acid which ursodeoxycholic acid took up increased, whereas those which cholic acid, chenodeoxycholic acid and deoxycholic acid took up were decreased.

Poliovirus infection of established human blood cell lines: Relationship between the differentiation stage and susceptibility or cell killing

The replication of type 1 poliovirus in 13 established human blood cell lines differing in the differentiation stage and cell lineage was investigated. Three T (CCRF-CEM, CCRF-HSB-2, and Molt-3) and three B (Raji, CCRF-SB, and RPMI 8226) cell lines showed no cytopathic effects (CPE) or virus production. CPE associated with virus production were detected in the other seven cell lines: HL-60, ML-1, and KG-1 (granulocytic lineage), U-937 and THP-1 (monocytic lineage), K-562 (erythroid lineage), and Molt-4 (T cell lineage). These susceptible cell lines greatly differed in the speed at which the CPE progressed. The progression of CPE was faster in relatively well-differentiated cell lines such as HL-60 and U-937, independently of the multiplicity of infection, than in less differentiated cell lines such as K-562, KG-1, and THP-1. Thus, for the same lineage, the speed at which CPE progressed became proportionally higher with subsequent differentiation stages. In the K-562 cell culture, CPE were not observed until at least 5 days postinfection (p.i.), while more than 80% of HL-60 cells were killed within 3 days p.i. There were no significant differences between infected HL-60 and K-562 cells in the efficiency of infection determined at 8 hr p.i. by the indirect immunofluorescent technique, the rate of virus growth, or the amount of viral capsid protein synthesized. This indicated that there were similar viral replication cycles in the two cell lines. These observations suggest that the killing function of the virus is expressed more slowly in K-562 cells than in HL-60 cells.

Uneven distribution of enzymatic alterations on the liver cell surface in experimental extrahepatic cholestasis of rat.

Abstract The effect of bile duct ligation on enzyme activities in the subfractions of the rat liver plasma membrane was investigated. Two subfractions were isolated from the rat liver plasma membrane by homogenization and subsequent centrifugation in a discontinuous sucrose gradient. The light subfraction contained fragments of the bile canalicular surface and the heavy fraction contained fragments of the sinusoidal and lateral surfaces of the hepatocyte. Bile duct ligation decreased NaK ATPase and Mg-ATPase activity in the light subfraction, whereas it had no significant effect on these enzyme activities in the heavy fraction. Leucyl-β-naphthylamidase activity was reduced and alkaline phosphatase activity was increased in both subfractions. These facts suggested that ligation of the bile duct led to loss of the secretory polarity of the liver cell. The in vitro effects of some bile acids on the membranebound enzymes in the light subfraction were investigated, and a possible involvement of chenodeoxycholic acid in the alteration of enzyme activities in the bile canalicular membrane was suggested.

Naturally occurring anti-interferon-α2a antibodies in patients with acute viral hepatitis

The occurrence of antibodies against recombinant human interferon‐α2a (IFN‐α2a) in patients with acute viral hepatitis (AVH) was examined by ELISA. Naturally occurring IgG anti‐IFN‐α2a were found in 50% of patients with type A, 50% of those with type B and in 8.3% of those with non‐A, non‐B AVH. The corresponding frequencies of IgM antibodies were 80%, 30% and 33.3%, respectively. IgM anti‐IFN‐α2a were found more frequently in patients with AVH type A than in normal control subjects (P < 0.01). Anti‐IFN‐α2a were detectable at the highest frequency 3 weeks after acute onset and then became negative. An absorption experiment revealed that IgM anti‐IFN‐α2a did not cross‐react with recombinant human IFN‐α2b. Immunoblotting analysis confirmed the binding of antibodies to IFN‐α2a. Sera positive for IgG and/or IgM anti‐IFN‐α2a were unable to neutralize IFN‐α2a. The appearance of anti‐IFN‐α2a was not correlated with disease severity. There was no evidence to suggest that anti‐IFN‐α2a impaired the elimination of hepatitis virus. This is the first study to demonstrate the occurrence of anti‐IFN‐α2a in patients with AVH. Detection of anti‐IFN‐α2a may be useful for clarifying any underlying immune events in various diseases.

Characterization of Rose Bengal binding to sinusoidal and bile canalicular plasma membrane from rat liver

The binding of Rose bengal, a model organic anion, to sinusoidal and bile canalicular membrane fractions isolated from rat liver was compared. The fluorescence change of Rose bengal after being bound to liver plasma membranes was utilized for measuring the binding. The dissociation constants (Kd = 0.1-0.12 microM) and the binding capacities (n = 11-15 nmol/mg protein) for Rose bengal are comparable between the two membrane fractions, although the n value for sinusoidal membrane is somewhat larger than that for bile canalicular membrane. The Rose bengal binding to both membrane fractions was inhibited by various organic anions at relatively low concentrations, i.e., the half-inhibition concentrations (IC50) for Indocyanine green, sulfobromophthalein, Bromophenol blue and 1-anilino-8-naphthalene sulfonate were 0.1, 100, 1.5-2.5 and 100 microM, respectively, while taurocholate did not inhibit the Rose bengal binding to either membrane fraction at these low concentration ranges. The type of inhibition of sulfobromophthalein and Indocyanine green for Rose bengal binding is different between the two membrane domains. That is, in sinusoidal and bile canalicular membrane fractions, these organic anions exhibit mixed-type and competitive-type inhibition, respectively. It was suggested that the fluorescence method using Rose bengal may provide a simple method for detecting the specific organic anion binding protein(s) in the liver plasma membrane.

Antibodies against sulphatide in sera from patients with autoimmune rheumatic diseases

We tested sera of patients with various autoimmune rheumatic diseases for the presence of antibodies against sulphatide (an acidic glycosphingolipid), identified as a target antigen for antibodies against the liver cell membrane. Thirty‐five percent (7/20) of patients with lupus in the active stage possessed anti‐sulphatide antibodies, whereas 10% (2/20) of those in the inactive stage and 20% (4/20) of those in the stationary stage possessed such antibodies. Moreover, 10%. (3/29) of patients with other autoimmune rheumatic diseases also possessed anti‐sulphatide antibodies. The level of anti‐sulphatide antibodies was significantly correlated with the levels of anti‐double‐stranded (ds) DNA antibodies (r = 0.634, P <0.001) and dextran sulphate‐binding IgG (r = 0.407, P < 0.001). The serum levels of antibodies against sulphatide were correlated with a history of seizures or psychosis in patients with autoimmune rheumatic diseases. Gels coupled with polyanionic dextran sulphate, monoanionic sulphanilic acid and DNA were shown effectively to adsorb anti‐sulphatide antibodies in the sera of patients with active systemie lupus erythematosus (SLE) and autoimmune chronic active hepatitis (AI‐CAH). These results suggest that the observed reactivity with sulphatide is due to the presence of antibodies capable of reacting with various anionic molecules in the sera of patients with autoimmune rheumatic diseases as well as those with AI‐CAH.

Down-regulation of prostaglandin E2 receptors in regenerating rat liver and its physiological significance.

The properties of prostaglandin (PG) E2 receptors in regenerating liver were studied using rat hepatocytes in primary culture. The control cells possessed stereo-specific PGE2 receptors with Bmax and Kd values, at 4 degrees C, of 526 fmol/mg protein and 6.5 nM respectively. In cells from regenerating liver after 70% hepatectomy, Bmax was reduced to 42-43% that of the controls; Kd did not change. Administration of indomethacin before surgery prevented Bmax reduction. These results indicate that PGE2, produced during the regeneration process, evoked cellular events and regulated the density of its receptors.

Abnormal high density lipoprotein of primary biliary cirrhosis analyzed by high performance liquid chromatography

The lipoprotein profile of patients with primary biliary cirrhosis (PBC) has been assessed by high performance liquid chromatography (HPLC). Without exception, all subjects with PBC had an abnormal high density lipoprotein (HDL), which was larger than normal HDL. The elution time from an HPLC column of the abnormal HDL separated it clearly from normal HDL. The HDL of subjects with asymptomatic and symptomatic PBC could also be distinguished. The HDL of asymptomatic subjects was eluted between that of normal and symptomatic subjects. The lipid and protein composition of abnormal HDL was characterized by an increase in phospholipid and a decrease in protein, resulting in an increase in the ratio of phospholipid/protein. This ratio was higher in symptomatic than in asymptomatic PBC. Incubation of dimyristoyl phosphatidylcholine liposomes with normal serum resulted in the formation of an altered high density lipoprotein. These findings suggest that an overload of phospholipid may play a role in the formation of the abnormal HDL in primary biliary cirrhosis.

Effects of teleocidin on the morphology and c-myc expression of hepatoma cells which are not inhibited by protein kinase antagonists

PLC/PRF/5 hepatoma cells cultured with a tumor promoter teleocidin showed polygonal cellular appearance with many vacuole-like structures, and reduced both c-myc mRNA level and growth rate. These teleocidin effects were partly mimicked by sodium butyrate but not by a protein kinase C stimulant 1-oleoyl-2-acetylglycerol(OAG). Protein kinase C inhibitor 1-(5-isoquinolinyl-sulfonyl)-2-methyl-piperazine(H7), calmodulin-dependent protein kinase antagonist N-(6-aminohexyl)-5-chloro-1-naphthalene sulfonamide(W7) and topoisomerase II inhibitor novobiocin failed to inhibit the effects of teleocidin. These results may suggest the presence of still unknown biochemical pathways which mediate the actions of teleocidin.