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Biomedical subjects

T Kurimura

Publications and source records attributed to T Kurimura.

At least 37 records · Page 2Linked to original sources
PubMed1990

A new method for extracting DNA or RNA for polymerase chain reaction.

The use of glass powder suspension for the extraction of RNA or DNA was studied to simplify the procedures of polymerase chain reaction (PCR). Using this procedure, proviral DNA of human T-lymphotropic virus type-1 (HTLV-1) in the blood of an asymptomatic virus carrier and viral RNA of human immunodeficiency virus (HIV) in the blood of an AIDS patient were easily detected by PCR employing glass powder. The use of glass powder is a simple and highly efficient procedure for the extraction of DNA or RNA, and can be applied for routine PCR.

Acquired Immunodeficiency Syndrome↗
PubMed1990

The role of the vimentin intermediate filaments in rat 3Y1 cells elucidated by immunoelectron microscopy and computer-graphic reconstruction.

The three-dimensional arrangement of vimentin intermediate filaments (IF) was studied in 3Y1, rat fibroblastic cell line, to elucidate its biological role in the cell. While actin filaments were observed exclusively in the superficial part of the cell, vimentin IF were found to be abundantly present in the inside of the cell where microtubules were occasionally discovered. By whole-mount immunoelectron microscopy and computer-graphic reconstruction of serial thin sections, it was observed in more detail that vimentin IF are located very close to the nucleus, endoplasmic reticulum, and mitochondria. Vimentin IF were observed to be attached to these organelles laterally or terminally. Thus, we can reasonably assume that vimentin IF are major cytoskeletal structures deep inside the cell and that they play an important role in supporting the location of the organelles. This is the first report which has visualized the three-dimensional relationship between vimentin IF and the organelles of the cell.

Animals↗
PubMed1990

Variations in growth capacity of HIV in peripheral blood mononuclear cell preparations from different individuals.

To investigate whether there are variations in the growth capacity of HIV in peripheral blood mononuclear cell (PMBC) preparations from different individuals, PBMC cultures prepared from 27 healthy donors were infected with the fresh HIV-1 isolates JH/3 or JMH/1. After infection of the PBMC with HIV, the culture fluid was checked for infectivity (TCID50) and the level of p24 antigen. Significant diversity was observed in these values in different individuals. No correlation between the percentage of CD4-positive lymphocytes or the growth ability of PBMC and TCID50 was shown. Whether PBMC was infected with JH/3 or JMH/1 also made no difference. In some cases, however, the culture fluid possessed high (or low) p24 levels despite a low (or high) infectious virus yield. The E-rosette-positive cell fraction was separated from the PBMC preparations which showed the highest and the lowest virus yield, and the yield was determined after infection with JMH/1. The difference in virus yield and the TCID50/p24 ratio between these donors was also observed in these T-cell-enriched cell cultures. The results suggest that variations in the growth capacity of HIV in different PBMC preparations are due to the T-cell population and that the source of the PBMC influences not only the binding of the virions onto the cell surface but also the replication of the virus in the cell.

Adult↗
PubMed1990

Synthesis and processing of a gp28/32 membrane glycoprotein induced by Marek's disease virus serotype 2.

The post-translational events leading from the precursor to the processed forms of a glycoprotein with an Mr of 28K to 32K (gp27/32) of Marek's disease virus (MDV) serotype 2 were examined with pulse-chase experiments and treatment with tunicamycin and monensin. Cell-free translation of infected cell mRNA followed by immunoprecipitation analysis suggested that a polypeptide with a size of 22K is the initial precursor. Experiments with endo-beta-N-acetylglucosaminidase H and endo-beta-N-acetylglucosaminidase F indicated that gp28/32 contains mostly N-linked oligosaccharides of the complex type. These studies showed that 22K, the initial product, is then processed through intermediates to the 28K to 32K form.

Electrophoresis, Polyacrylamide Gel↗
PubMed1990

Disappearance of anti-p17 correlates with successful isolation of human immunodeficiency virus and deterioration in clinical status.

Anti-p17 was examined as a human immunodeficiency virus (HIV) marker predicting the onset of the acquired immunodeficiency syndrome (AIDS). Two different comparisons were done: (1) anti-p17 negativity and successful HIV isolation; and (2) anti-p17 negativity and clinical status, using the Centers for Disease Control classification. Anti-p17 negativity was not only significantly correlated with deterioration in clinical status (P less than 0.01), but also with successful HIV isolation (P less than 0.05). HIV isolation was affected by several drugs, e.g. zidovudine. However, the results of the antibody test were not affected. It is significant that anti-p17 can compensate for the defects of virus isolation.

Acquired Immunodeficiency Syndrome↗
PubMed1990

Heterogeneity of Hanganutziu-Deicher antigen glycoproteins in different species animal sera.

A heterophilic Hanganutziu-Deicher (HD) antigen is present in many animal sera except human and chicken sera. To visualize the antigenic molecules, nine species animal and human sera were analyzed by SDS-PAGE, followed by Western blotting and immunostaining with avain anti-N-glycolylneuraminyl-lactosyl ceramide antibody which recognizes the terminal N-glycolylneuraminic acid moiety of glycoconjugates as an epitope of the HD antigen. Several HD antigen-active glycoprotein bands were detected in the sera of fetal calf, calf, horse, goat, monkey, rabbit, guinea pig, rat and mouse, except for human serum. The HD antigenic proteins showed heterogeneities in their molecular weights and were not identical with any major band visualized with silver-staining, indicating that they are minor components of serum proteins in each animal. Neuraminidase treatment destroyed the antigenicity of all proteins, confirming that N-glycolylneuraminic acid (NeuGc) at the non-reducing terminal of carbohydrate chains is the antigenic epitope in serum glycoprotein molecules as already confirmed in glycosphingolipid (GSL) antigens. The finding of HD-antigenic glycoproteins in animal sera suggests that they also stimulate HD antibody production in patients who received animal antiserum for therapeutic aim.

Animals↗
PubMed1990

Budding process and maturation of human immunodeficiency virus examined by means of pre- and post-embedding immunocolloidal gold electron microscopy.

The techniques of pre- and post-embedding immunocolloidal gold electron microscopy were tested for their usefulness for analyzing the morphogenesis of human immunodeficiency virus (HIV) strain LAV employing U-937 cells persistently infected with the virus (U-937/LAV). By both techniques the following results were obtained. (1) Budding process was restricted to a localized area at the membrane adjacent to the Golgi apparatus. (2) The distribution of viral envelope antigens was restricted to the electron-dense crescent structures on the cell surface. (3) Virions released from the cells could be classified mainly into 2 categories: virions with doughnut-shaped cores or with conical cores at the center of the particles. And p24 proteins localized on the thick electron-dense cores of both types. These findings support the idea that pre- and post-embedding immunogold electron microscopy is very useful for studying the morphogenesis of viruses.

HIV↗
PubMed1989

Nucleotide sequences of gag and env genes of a Japanese isolate of HIV-1 and their expression in bacteria.

A DNA clone of HIV-1, JH3, was isolated from a Japanese patient with hemophilia and the gag and env genes were sequenced. The nucleotide and deduced amino acid sequences were similar to those reported and showed high divergence in the env gene, particularly in the extracellular domain of the env. The genetic variation of JH3 isolated from a Mongolian was within the range of those of isolates from whites and blacks. The gag and env polypeptides were efficiently expressed in E. coli as fusion proteins with beta-galactosidase, and the products were shown to be useful as diagnostic reagents.

Amino Acid Sequence↗
PubMed1989

[HBV carriers and HDV superinfection--studies on the cases of HBsAg clearance].

We studied the annual clearance rates of hepatitis B surface antigen (HBsAg) and the annual seroconversion rates of HBsAg (HBs seroconversion rates), and the correlation between HBsAg clearance and hepatitis delta virus (HDV) superinfection in hepatitis B virus (HBV) carriers in Japan. Out of 1,029 HBV carriers followed for more than 36 months, 56 cases were cleared of HBsAg from the sera, and 24 of these cases developed hepatitis B surface antibody. The annual clearance rate of HBsAg was 0.94% and the annual HBs seroconversion rate was 0.27%. These rates increased with aging, especially above 30 years of age. Antibody to HDV was detected in three cases with increased serum alanine aminotransferase activity preceding HBsAg clearance. These data indicate that HDV superinfection may play a role in induction of the HBsAg clearance in HBV carriers in Japan.

Adult↗
PubMed1989

[Inhibitory effect of inositol hexasulfate and inositol hexaphosphoric acid (phytic acid) on the proliferation of the human immunodeficiency virus (HIV) in vitro].

The monosaccharide substances inositol hexasulfate (IHS) and inositol hexaphosphoric acid (Phytic acid, IHP) were investigated for their antiviral effect on the human immunodeficiency virus (HIV) in vitro. In MT-4 cells IHS completely inhibited the cytopathic effect of HIV and the HIV specific antigen expression at a concentration of 1.67 mg/ml. IHP moderately inhibited both of HIV effects as mentioned above.

Antiviral Agents↗
PubMed1989

[Occurrences of human papilloma virus DNA in cervical carcinomas].

Out of 16 cases involving a cervical carcinoma that were investigated by Southern blot hybridization, found were human papilloma virus (HPV) types 16 and 18 DNA sequences in 8 (50%) and in one (6.3%), respectively. Six out of the 8 HPV 16-positive specimens were from squamous cell carcinomas, one was from an adenocarcinoma, and the remaining specimen was from an argyrophil small cell carcinoma. In 7 out of 9 HPV-positive specimens, the viral sequences were integrated in the tumor cell genome, whereas in the remaining two they were not integrated and remained circular and/or oligomeric in form.

Adenocarcinoma↗
PubMed1988

Influence of hepatitis delta virus superinfection on the clearance of hepatitis B virus (HBV) markers in HBV carriers in Japan.

To elucidate the influence of hepatitis delta virus (HDV) superinfection on the clearance of hepatitis B virus (HBV) associated antigens in HBV carriers, we examined for antibody to hepatitis delta antigen (anti-HD) serial sera collected from 1,029 HBV carriers in Kure, Japan. Of the 242 HBV carriers with hepatitis B e antigen (HBeAg), 28 became seropositive for anti-HD, of whom 18 (64.3%) cleared HBeAg; 214 did not become seropositive for anti-HD, of whom 70 (32.7%) cleared HBeAg. Thus, HBeAg clearance was observed in a significantly higher proportion of HDV-superinfected carriers as compared with carriers without HDV infection (P less than 0.005). In the 56 HBV carriers who cleared hepatitis B surface antigen (HBsAg), anti-HD was detected in three cases with increased serum alanine aminotransferase activity preceding HBsAg clearance. The duration of anti-HD seropositive state was less than 5 years, and the titer of anti-HD was relatively low in every case. These data suggest that the HDV infection rate in Japan is higher than previously reported, that HDV superinfection can be one of the factors that induce the HBeAg clearance and HBsAg clearance in HBV carriers, and also that the most likely outcome of HDV superinfection in HBV carriers in Japan may be acute self-limited infection.

Adult↗
PubMed1988

Inhibition of growth of HIV by human natural interferon in vitro.

The effect of human natural interferons (IFN) alpha, beta, and gamma on the replication of human immunodeficiency virus (HIV) strain LAV in T cell lines TALL-1 and CCRF-CEM and peripheral blood lymphocytes (PBL) was studied. The growth of TALL-1 was moderately sensitive to these IFN, whereas that of CCRF-CEM was resistant to them. The progeny virus yield of LAV in TALL-1 at the time of its peak was reduced to 10% of the control level at IFN-alpha, beta, and gamma concentrations of 3, 11, and 23 IU/ml, respectively. These concentrations of IFN-alpha, beta, and gamma did not affect the cell growth. In CCRF-CEM, IFN-alpha, beta, and gamma at the concentration of 50, 60, and 76 IU/ml reduced the progeny virus to 10% of the control level. The virus growth in PBL was more sensitive to IFN than that in CCRF-CEM. The progeny virus yield was reduced to 10% of the control level by IFN-alpha and beta concentrations of 5 IU/ml and less than 5 IU/ml, respectively.

Cell Division↗
PubMed1988

Replication of human immunodeficiency virus in two T-cell lines and their application as antigens for immunofluorescence.

Two T-cell lines, TALL-1 and CCRF-CEM, were infected with human immunodeficiency virus (HIV), strain LAV, to explore the time course of the appearance of various virus specific antigens, and to establish an antibody assay system by indirect immunofluorescence (IF). These cells were infected with LAV at two different input multiplicity of infection (MOI). Antigens were tested by Western blot analysis (WB) and IF. Antigens for WB were extracted from the infected cells at various times after infection, but pooled sera of American HIV carriers could not recognize gp41 or gp160. Antigen expression was highest in CCRF-CEM, but, as the antigen for IF, TALL-1 infected at the MOI of 8.0 was the most suitable 7 days after infection, because it includes a fairly large number of uninfected cells, which served as the internal control.

Cell Line↗