Studies on anti-LP-1 and anti-Tamm-Horsfall glycoprotein in chronic liver disease using ADCC assay against antigen-coated target cells.

To study autoantibodies against liver cell surface membrane clinically, anti-LP-1 and anti-Tamm-Horsfall glycoprotein (THGP) were determined in the sera of patients with various liver diseases. They were detected by ADCC assay using antigen-coated cells as the target. A high incidence of anti-LP-1 was seen in chronic hepatitis (CH), liver cirrhosis (LC), primary hepatic cancer with cirrhosis (PHC), and primary biliary cirrhosis. The incidence of anti-THGP was also high in CH, LC, and PHC. Both anti-LP-1 and anti-THGP were detected in 2 of 3 patients with lupoid hepatitis. The patients studied here had no obvious evidence of renal tubular acidosis or pyelonephritis. Serum alanine transaminase activity, serum gamma-globulin content, and the presence of rheumatoid factors were not associated significantly with the presence of anti-LP-1 or anti-THGP in chronic liver disease. In 7 cases of CH tested serially during their clinical course, anti-LP-1 and/or anti-THGP tended to appear during acute exacerbations. The demonstration of anti-LP-1 and anti-THGP suggested that their appearance was related to the development of chronic liver disease.</p

Metabolic fates of isovaleric acid and isovalthine in rats

1. Isovaleric acid-1-C14, -4-C14, or C14-CaC03 with or without non-isotopic isovaleric acid was orally administered to rats and the incorporation of these isotopes into liver cholesterol, fatty acid, or urinary isovalthine was examined. 2. Isopropyl group of isovaleric acid was more efficiently utilized for cholesterol synthesis than carboxyl group, and also for cholesterol synthesis than for fatty acid. These results indicate that isovaleric acid is cleaved into two fragments before it is utilized for cholesterol synthesis. 3. Carbon dioxide was used for the synthesis of liver cholesterol and of liver fatty acid. Isovaleric acid seems to enhance the incorporation of carbon dioxide into cholesterol. 4. All the experimental rats received isotopic or non-isotopic isovaleric acid excreted isovalthine, but no radioactivity was found in it. Thus, isovaleric acid residue of urinary isovalthine molecule is not derived from isovaleric acid administered, and carbon dioxide is not the carbon source of urinary isovalthine. 5. Suspicious metabolism of isovaleric acid or of carbon dioxide was discussed. 6. Isotopic isovalthine which was synthesized from (± ) &#945;-bromoisovaleric acid-4-C14 is administered to rat and it was found that the isotope did not incorporate into cholesterol or fatty acid of liver and of brain. About 15% of isotopic isovalthine was recovered in urine up to the next day after injection. The large part of isovalthine was missing.</p

Detection of negative strand RNA of hepatitis C virus in infected liver and serum.

The negative strand RNA of hepatitis C virus, supposed to be a replicative intermediate of the virus appears to indicate viral replication. In this study, we detected the negative strand RNA by using reverse transcription-polymerase chain reaction with RNase A digestion to degrade the remaining positive strand genomic sequence of the virus after complementary DNA (cDNA) synthesis. In vitro transcribed positive-stranded mutant RNA was not detected by this method. Sample sera and liver tissues of 16 patients with chronic hepatitis C virus infection (liver fibrosis, 1; chronic hepatitis, 13; liver cirrhosis, 2) were analysed for negative strand RNA of hepatitis C virus. The negative strand RNA sequence was detected in 15 (93%) of 16 liver tissues and in 11 (78%) of 14 sera. The study demonstrated that negative strand RNA of hepatitis C virus in serum and liver tissue could be specifically detected.</p

SEISMIC RETROFITTING METHOD FOR STEEL STRUCTURES BY KNEE BRACES JOINTED BY HIGH HARDNESS VISES

The authors have developed a new method of seismic retrofit for steel structures by adding knee braces which is jointed to the existing structures by vises with screw bolts made of high hardness metal. The slipping strength of the connection had been studied through axial tensile test of steel coupons jointed by the vises. The features of the connection by the vise are easy setup and providing the strength as much as by the normal bolts joint. An experimental study had been conducted for the specimens of steel frames retrofitted by knee braces. The knee brace was jointed to the steel frames by the vises. Two types of failure modes were investigated. One was slipping behavior at the connection and the other was bucking behavior at the knee braces. The strengths of the specimens were estimated by the simple calculations for the two types of failure modes

EFFECTS OF PLAYING BADMINTON ON BONE PROPERTIES USING CALCANEAL QUANTITATIVE ULTRASOUND: A PRELIMINARY STUDY

Purpose: This study was designed to investigate the effects of playing badminton on calcaneal bone properties. Methods: Eleven sedentary collegiate women were recruited. They played badminton for 75 min, 2 days per week, for 10 weeks. The right calcaneus was assessed to measure speed of sound and broadband ultrasound attenuation using a quantitative ultrasound device. A stiffness index was derived from both the speed of sound and broadband ultrasound attenuation. Results: After the training period, broadband ultrasound attenuation and stiffness index did not change significantly, whereas speed of sound significantly increased, Conclusion: The results indicate that playing badminton influences calcaneal bone properties in a positive manner.  Article visualizations

Genetic variation of putative core gene in hepatitis C virus.

Genetic variation of hepatitis C virus was assessed. We prepared RNA fractions from 21 patients' sera which were positive for hepatitis C virus RNA, synthesized their cDNAs, and amplified fragments, 406 base pairs, encoding a putative core protein, by polymerase chain reaction. One of them, N 15, was cloned and sequenced. N 15 showed 92.4% homology at the nucleotide level and 97.0% homology at the amino acid level compared with HC-J 1 which is the first isolated clone in Japan and similar to that isolated in USA. By restriction fragment length polymorphisms analysis, 14 out of 21 patients (66.7%) showed the same pattern as N 15. No patients showed the pattern of HC-J 1. We could not find a correlation between the genetic variation and clinical features of hepatitis C virus infection. These results indicate that the region, which encodes the core protein and is believed to be relatively conserved in hepatitis C virus genome, has several variations at the nucleotide level, and the major part of hepatitis C virus in Okayama district is different from HC-J 1 and the USA clone.</p

Development of syngeneic monoclonal anti-idiotype antibodies to mouse monoclonal anti-asialoglycoprotein receptor antibody.

Anti-idiotype antibodies (Ab2) play an important role in the homeostasis of immune responses and are related to the development and the disease activity of certain autoimmune diseases. The asialoglycoprotein receptor (ASGPR) is considered one of the target antigens in the pathogenesis of autoimmune chronic active hepatitis (AIH). We previously developed a mouse monoclonal antibody (clone 8D7) which recognizes rat and human ASGPR. In this study, to help investigate the anti-ASGPR antibody-anti-idiotype antibody network in patients with AIH, we developed a syngeneic mouse monoclonal Ab2 to the 8D7 anti-ASGPR antibody (Ab1). One clone, designated as 3C8, tested positive for specific reactivity to 8D7-Ab1 and did not bind to other irrelevant immunoglobulins. By competitive inhibition assays, the binding of 8D7-Ab1 to liver membrane extracts, i.e., the crude antigen preparation, was inhibited by 3C8-Ab2 in a dose-dependent manner, and the binding of 8D7-Ab1 to 3C8-Ab2 was inhibited by the liver membrane extracts. In the immunohistochemical analysis, 3C8-Ab2 blocked the specific staining of sinusoidal margins of rat hepatocytes by 8D7-Ab1. These results suggest that 3C8 anti-idiotype antibody recognizes the specific idiotypic determinants within the antigen-binding site of 8D7-Ab1.</p

Hepatitis C virus quasispecies in cancerous and noncancerous hepatic lesions: the core protein-encoding region.

We have shown that highly proofreading DNA polymerase is required for the polymerase chain reaction in the genetic analysis of hepatitis C virus (HCV). To clarify the status of HCV quasispecies in hepatic tissue using proofreading DNA polymerase, we performed a genetic analysis of the HCV core protein-encoding region in cancerous and noncancerous lesions derived from 4 patients with hepatocellular carcinoma. In contrast to the previously published data, we observed neither deletions nor stop codons in the analyzed region and no significant difference in the complexity of HCV quasispecies between cancerous and noncancerous lesions. This result suggests that the HCV core gene is never structurally defective in hepatic tissues, including cancerous lesions. However, in 3 of the patients, the consensus HCV species differed between cancerous and noncancerous lesions, suggesting that the predominant replicating HCV species differs between these 2 types of lesions. Moreover, during the course of the study, we obtained several interesting variants possessing a substitution at codon 9 of the core gene, whose substitution has been shown to induce the production of the F protein synthesized by a - 2/+1 ribosomal frameshift.</p

Quantitation of Hepatitis C Virus RNA in Liver Tissue as a Predictive Marker of the Response to Interferon Therapy in Chronic Hepatitis C

Recently, factors predicting the response to interferon (IFN) therapy against hepatitis C virus (HCV) have received much attention. To evaluate the usefulness of the quantitation of intrahepatic HCV RNA as a predictive marker of the response to IFN therapy, we compared the amount of intrahepatic HCV RNA with serum levels in 16 patients. Eleven patients who had 10(10) copies/g or more of intrahepatic HCV RNA had increased level of serum alanine aminotransferase (ALT) after IFN therapy, while 4 of 5 patients who had less than 10(10) copies/g of intrahepatic HCV RNA achieved sustained normalization of serum ALT level and were designated as complete responders. Four complete responders possessed significantly less HCV RNA in the liver parenchyma than partial and nonresponders (P = 0.010, Mann-Whitney U-test), but the amount of HCV RNA in the serum was not significantly different between those groups. In conclusion, the results suggest that the quantitation of intrahepatic HCV RNA is a better indicator of the response to IFN therapy than serum HCV RNA.</p