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Biomedical subjects

O Chisaka

Publications and source records attributed to O Chisaka.

27 records · Page 2Linked to original sources
PubMed1989

Detection of hepatitis B virus X gene protein and antibody in type B chronic liver disease.

The genome of the hepatitis B virus contains a sequence (X gene) whose role is unclear. The almost complete region of the hepatitis B virus X gene was expressed in Escherichia coli, with the resulting protein being approximately 17 kilodaltons in molecular weight. Sera from 139 subjects were analyzed by Western blot analysis. Of the hepatitis B surface antigen-positive patients, anti-X was not found in 4 patients with acute hepatitis and in 12 healthy carriers, but was present in 41% (21/51) of the patients with chronic hepatitis, 63% (15/24) of those with liver cirrhosis, and 46% (12/26) of those with hepatocellular carcinoma. The expression of the X product in the liver tissues (43 hepatitis B surface antigen-positive patients) was investigated using an indirect immunohistochemical method. The X protein was observed in 64% (21/33) of the patients with chronic hepatitis and 50% (5/10) of those with liver cirrhosis, and was found when the serum was negative for anti-X. Hepatitis B core antigen was frequently expressed together with the X protein in the liver. The conclusions reached were that the frequency of anti-X increases with the length of chronic hepatitis B virus infection, that anti-X may suppress the expression of the X protein in the liver, and that the X protein may be related to hepatitis B virus replication.

Blotting, Western↗
PubMed1989

An in vitro system for screening anti-hepatitis B virus drugs.

A human hepatoblastoma cell line (HB 611) that continuously synthesizes hepatitis B viral (HBV) DNA was grown in the presence of various inhibitors of DNA synthesis, and the DNA from the cells was analyzed by the Southern blotting method to examine selective inhibition of the viral DNA synthesis. Among those that showed selective inhibition, and interferons alpha and beta, acyclovir, and dideoxy cytidine were effective. This system should be useful for screening new antiviral agents against HBV.

Acyclovir↗
PubMed1989

Expression and replication of hepatitis B virus genome in transgenic mice.

We produced transgenic mice by microinjecting a partial tandem duplication of the complete hepatitis B virus (HBV) genome into fertilized eggs of C57BL/6 mice. One of eight transgenic mice was a high producer for HBV surface antigen (HBsAg) and HBV e antigen (HBeAg) in the serum. The HBV genomes were transmitted to the next generation and these F1 mice also produced HBsAg and HBeAg. mRNAs of 3.5, 2.1, and 0.8 kilobases were detected in the livers and the kidneys of these mice. In addition, a 0.8-kilobase RNA was detected in the testis. Single-stranded and partially double-stranded HBV DNAs were shown to be produced in the cytoplasm of the liver and kidneys. These HBV DNAs were associated with the core particles, indistinguishable from nucleocapsid produced in an infected human liver. Viral genome DNA was detected in the serum. These results demonstrate that the HBV genome integrated into the mouse chromosome acted as a template for viral gene expression, allowing viral replication. Thus, these transgenic mice should be useful for detailed studies of the replication and expression of HBV and for pathological studies of hepatitis, including the development of hepatocellular carcinoma.

Animals↗
PubMed1987

Purification of hepatitis B virus gene X product synthesized in Escherichia coli and its detection in a human hepatoblastoma cell line producing hepatitis B virus.

A fused gene containing 94% of the hepatitis B virus (HBV) open reading frame X was expressed in Escherichia coli, and its 17-kDa product was purified by ion-exchange chromatography. Antibody elicited against the X-gene product reacted with materials proximal to the nuclear membrane of a human hepatoblastoma cell line producing HBV particles. No such reaction was observed with the same cell line that did not produce HBV particles.

Carcinoma, Hepatocellular↗
PubMed1986

Cloning a defined region of DNA using a limited action of DNA polymerase: application to dissection of hepatitis B virus surface antigen gene.

Using dodecadeoxynucleotides as primers for DNA synthesis and 3'-o-chlorophenyl-phosphorylated dodecadeoxynucleotides as "stoppers" for chain elongation, pre-defined regions of a gene previously cloned in M13 single-stranded (ss) DNA phage were converted into double-stranded (ds) DNA utilizing the action of the Klenow fragment of Escherichia coli DNA polymerase I (PolIk). The resulting ds DNA was freed from the ss region by S1 nuclease treatment. This method can be used to obtain DNA fragments of any size with pre-defined 5' and 3' ends. About 15% of the input ss DNA template molecules are converted into ds DNA fragments. This technique was used to synthesize several DNA fragments from different portions of the hepatitis B virus surface antigen (HBsAg) gene. The products were then ligated into a yeast plasmid vector that carries the E. coli lacZ gene which is located downstream from the yeast acid-phosphatase promotor. Using this system, several fragments of HBsAg were produced in the form of beta-galactosidase fused protein.

Bacterial Proteins↗
PubMed1985

Expression of a synthetic human growth hormone gene in yeast.

A synthetic human growth hormone (hGH) gene was efficiently expressed under the control of the repressible acid phosphatase promoter in yeast (Saccharomyces cerevisiae). More than 10(6) molecules of hormone were formed per cell despite the fact that the gene was constructed with codon preference for Escherichia coli.

Alkaline Phosphatase↗
PubMed1984

Expression of poliovirus complementary DNA coding for viral antigenic determinants in Escherichia coli.

DNA sequences coding for the immunogenic capsid protein VP1 and/or VP3 of poliovirus strain LSc-2ab (Sabin 1) were prepared by digesting the cloned complementary DNA with restriction endonuclease PstI. The DNA fragments were inserted into the unique PstI site of Escherichia coli plasmid vectors pBR322, pKT 280 and/or pKT 287 that lay in the region expressed under control of the penicillinase promoter system. In the expression vectors, poliovirus sequences were designed to be read in phase and therefore to be expressed as fusion proteins with the bacterial peptides. In addition, the Escherichia coli tryptophan operon promoter-operator system was inserted upstream of the penicillinase system to obtain stronger expression of the poliovirus sequences. Escherichia coli transformed with these plasmids appeared to produce the antigenic polypeptides, which were detected by immunoprecipitation with antibodies to capsid proteins VP1 and/or VP3 followed by SDS-polyacrylamide gel electrophoresis.

Amino Acid Sequence↗
PubMedDate not supplied

Heat shock-induced homeotic transformations of the axial skeleton and associated shifts of Hox gene expression domains in mouse embryos.

Pregnant ICR mice were immersed in water at 42 degrees C for up to 15 min on Day 8.5 of gestation (plug day = Day 0), and their term fetuses were double stained with alcian blue and alizarin red S for skeletal examination. Heat exposure for 15 min induced homeotic vertebral transformations in more than one-third of the living fetuses, in which the morphologic identity of vertebrae (T6-S1) was shifted anteriorly by one or two segmental levels. The frequency of fetuses with vertebral transformations and the degree of the shift of vertebral identity were dependent on the length of heat exposure. The expression domains of Hoxa-7, Hoxc-8, and Hoxc-9 genes as examined by whole mount in situ hybridization were found to be shifted anteriorly in heated embryos. The heat-induced shifts of Hox gene expression domains were consistent with the observed vertebral transformations and suggested correlation or colinearity with the clustered organization of the Hox genes. The result of the present study indicates that a brief heat shock at a critical stage of differentiation can interfere with the normal establishment of Hox codes and subsequently, perturb the specification of vertebral identity.

Animals↗