PubMed Health⌕ Search

Biomedical subjects

T Takasaki

Publications and source records attributed to T Takasaki.

At least 19 recordsLinked to original sources
PubMed1997

Electron microscopic study of human immunodeficiency virus type 1 (HIV-1) core structure: two RNA strands in the core of mature and budding particles.

The basic structure of human immunodeficiency virus type 1 (HIV-1) has been investigated morphologically; however, the internal structure of HIV-1 core is not well understood. We studied the internal structures by transmission electron microscopy. We modified the method for electron staining of ultrathin sections and processed electron microscopic photographs using a computer. We confirmed that a mature HIV-1 particle had two copies of RNA strands in a cone-shaped core. These two RNA strands formed a coiling structure and interwound each other, and were already present in the late budding stage.

HIV-1↗
PubMed1996

Human antibodies responsible for binding inhibition and polymerization inhibition of human immunodeficiency virus type 1 reverse transcriptase.

Using a solid-phase non-radioisotopic (non-RI) reverse transcriptase (RT) assay, antibodies inhibiting human immunodeficiency virus type 1 (HIV-1) RT activity (RTI antibody) were investigated for their ability to inhibit binding of RT to a template-primer and DNA polymerization. The RTI antibody inhibited the binding of RT to the template-primer (BI antibody), and directly reacted with the RT-template-primer complex and inhibited enzymatic activity (PI antibody). The RTI antibody interfered with formation of the RT-template-primer complex suggesting that it recognized the antigenic site involved in template-primer binding of RT molecules. Since deoxynucleotide triphosphates (dNTPs) blocked inhibition of the RT activity by the PI antibody, the antigenic site recognized by the PI antibody may be closely related to the dNTP binding site. The seropositivities of the BI and PI antibodies were 84.6% and 91.2%, respectively, in HIV-1-infected individuals; healthy individuals, HTLV-I-positive individuals, autoimmune disease patients and leukemia patients were all seronegative. No significant correlation of residual RT activities was observed when BI and PI antibodies were compared (r = 0.688). It is possible that the epitopes recognized by the BI antibody differs from those recognized by the PI antibody. The assays described are able to detect BI and PI antibodies in the sera of HIV-1-infected individuals.

Animals↗
PubMed1996

Inhibitory effects of selenium, vitamin A and butylated hydroxytoluene on growth of human maxillary cancer cells in vitro.

The effects of vitamin A, selenium, and butylated hydroxytoluene (BHT) on the growth of a human maxillary cancer cell line were examined in monolayer cell cultures. The colony-forming assay showed a 50% reduction in the survival rate of the cell line at a concentration of 3.6 micrograms/ml of selenium, 28 micrograms/ml of vitamin A, and 74 micrograms/ml of BHT. Flow cytometric analysis with both FITC-labeled bromodeoxyuridine monoclonal antibody and propidium iodide demonstrated an increase of the S-phase fraction in the presence of selenium, an increase of the G0/G1-phase fraction in the presence of vitamin A, and an increase of the G2-M-phase fraction 1 day followed by an increase of G0/G1-phase fraction from the 3rd to 7th day when BHT was added. These results suggest that the mechanisms of inhibition of DNA synthesis by these compounds are different.

Antibodies, Monoclonal↗
PubMed1996

Recurrent laryngeal papillomatosis developing into laryngeal carcinoma with human papilloma virus (HPV) type 18: a case report.

We report a case of recurrent laryngeal papillomatosis which developed into laryngeal squamous cell carcinoma 11 years after the first diagnosis. Interestingly, we could identify HPV type 18 DNA in the carcinoma tissue using the polymerase chain reaction (PCR). Other known risk factors of irradiation, smoking, exposure to chemical agents, or a hereditary tendency to malignant tumours were not present in this case. Our finding suggests that HPV type 18 is another aetiological agent for laryngeal carcinoma.

Carcinoma, Squamous Cell↗
PubMed1996

Retrospective survey of chronic Q fever in Japan by using PCR to detect Coxiella burnetii DNA in paraffin-embedded clinical samples.

We used PCR to detect Coxiella burnetii DNA in paraffin-embedded tissues obtained from patients with chronic endocarditis in which the etiological agent had been unknown. On the basis of the published nucleotide sequence of the C. burnetii htpB gene, primers were chosen to produce an amplified fragment of 285 bp. A total of 60 samples from 56 patients were tested for the presence of C. burnetii DNA. Five samples from four patients were found to be positive. All of the amplified DNA fragments possessed a TthHB8I restriction site, as predicted from the published sequence of C. burnetii. In one of the four positive patients, rickettsia-like particles were found in sections of tissue stained by Gimenez's method. This is the first report of chronic Q fever in Japan.

Base Sequence↗
PubMed1996

[Clinical usefulness of trans-brachial angiography for abdominal diseases].

Trans-brachial angiography for abdominal disease has several advantages for the patient. For example, the patient can walk immediately following angiography and can return home or eat in the sitting position after examination. We performed trans-brachial angiography on 451 cases of abdominal disease from June 1992 to March 1995. The purpose of performing angiography in 215 of these cases was examination only, while that in the remaining 236 cases was embolization of liver tumor. Four hundred and five cases were used for the study of safety and accuracy, and the remaining 46 cases as well as another 51 cases of trans-femoral angiography were used for the study of technical difficulty. Based on these experience, we have concluded that trans-brachial angiography for abdominal disease is highly useful, safe and accurate, and can provide significant advantages for patients requiring abdominal angiography.

Adult↗
PubMed1995

Inhibitory effects of selenium, vitamin A and butylated hydroxytoluene on in vitro growth of human tongue cancer cells.

The effects of vitamin A, selenium and butylated hydroxytoluene (BHT) on the growth of a human tongue cancer cell line were examined in monolayer cell cultures. A colony-forming assay showed a 50% reduction in the survival rate of the cell line at a concentration of 2.6 micrograms/ml selenium, 60 micrograms/ml vitamin A and/or 38 micrograms/ml BHT. Relatively low concentrations of selenium markedly inhibited glucose consumption. Flow cytometric analysis with both fluorescein-isothiocyanate-labelled bromodeoxyuridine monoclonal antibody and propidium iodide demonstrated an increase in S-phase fractions 1 day after the addition of selenium, increased G0/G1 phase fractions in the presence of vitamin A and increased G2-M phase fractions when BHT was added. These results suggest that these compounds inhibit DNA synthesis of in vitro human tongue cancer cells by different mechanisms.

Butylated Hydroxytoluene↗
PubMed1995

Conversion of cathepsin E to enzymatic unstable form in gastric cancer cells.

A new acid proteinase in human gastric cancer, named medium moving proteinase (Med.P), was found also in a gastric cancer transplanted into nude mouse. However, Med.P disappeared when the samples prepared from gastric cancer tissues were left for over 4 weeks at -80 degrees C, whereas the activity of cathepsin E (CE) increased. When these samples were reduced by dithiothreitol (DTT), Med.P appeared again and the CE activity decreased. These phenomena, revealed by electrophoretic analyses, indicated that Med.P is a monomeric form of CE (mono-CE). At weakly alkaline pH and after heating, mono-CE appeared to be more unstable than CE. These results indicated that CE assume an enzymatically unstable monomeric form in cancer cells.

Animals↗
PubMed1994

Expression of p53 protein in benign epithelial hyperplasia, atypical ductal hyperplasia, non-invasive and invasive mammary carcinoma: an immunohistochemical study.

To clarify whether p53 protein expression is involved in multistep carcinogenesis or the progression of mammary ductal carcinoma, we investigated p53 protein expression in 83 invasive ductal carcinomas (IDC), 10 IDC with a predominant intraductal component, 13 non-invasive ductal carcinoma (NIDC), 16 atypical ductal hyperplasia (ADH) and 39 benign epithelial hyperplasia (EH), using immunohistochemistry. Expression of p53 protein was detected in 24 (28.9%) cases of IDC, 5 (50%) cases of IDC with a predominant intraductal component and 1 (7.6%) case of NIDC. No expression was observed in either ADH or EH. In IDC, including cases with a predominant intraductal component, p53 protein expression was associated with a higher histological grade (P < 0.0001) or mitotic index (P < 0.0005). Although overexpression of c-erbB-2 protein has also shown a similar association with these prognostic indicators, expression of p53 protein correlated regardless of the status of c-erbB-2 overexpression. Completely coordinated expression of p53 protein was seen in both intraductal and invasive components. The intraductal component in IDC including cases with a predominant intraductal component which expresses p53 protein had significantly higher histological grade (P < 0.0005) or more comedo-subtypes (P < 0.0001). These results suggested that p53 protein expression occurs at a stage of NIDC with high histological grade or in comedo-subtypes. Its expression is maintained throughout invasion.

Breast↗
PubMed1994

A high degree of homology exists between the protein encoded by SLG and the S receptor domain encoded by SRK in self-incompatible Brassica campestris L.

The sporophytic self-incompatibility system in Brassica campestris is controlled by multiple alleles of a single locus, S. We isolated and characterized cDNA clones that correspond to genes for S-glycoprotein (SLG) and S-receptor kinase (SRK) from an S9-homozygous strain. An SRK9 fragment was amplified by the polymerase chain reaction (PCR) by use of primers that corresponded to regions of the gene for the extracellular receptor domain (S-domain) and the kinase domain. The fragment was used as a probe to isolate an SRK9 cDNA clone from the cDNA library of stigmas of an S9-homozygote. An SLG9 cDNA clone was also isolated from the library by use of SLG8 cDNA as a probe. Restriction fragment length polymorphism (RFLP) linkage analysis of the F2 plants from an S8S9 hybrid demonstrated that the SRK and SLG genes were tightly linked to one another and were also linked to S-genotypes, as determined by pollination tests. The transcripts of SRK9 and SLG9 were detected in stigmas, but not in anthers or leaves. The cytoplasmic kinase domain encoded by SRK9 contained amino acid sequences that are conserved in serine/threonine kinases. The nucleotide sequence encoding the S-domain of SRK9 was 98.4% homologous to that of SLG9 at the nucleotide level. This observation suggests the existence of a mechanism for maintaining a high degree of similarity between SLG and the region that encodes the S-domain of SRK.

Base Sequence↗
PubMed1994

Electrophoretic analysis of a gastric cancer-associated acid proteinase using a highly sensitive detection system.

A highly sensitive detection system for acid proteinase separated on polyacrylamide gel was established. This system consisted of two-dimensional electrophoresis, combined with isoelectric focusing and polyacrylamide gel electrophoresis, and casein clotting (caseogram). Human urine, serum and gastric tissues obtained from normal individuals and gastric cancer patients were analyzed using this system. The previous electrophoretic method was not sufficiently sensitive to detected small amounts of pepsinogen (PG) C in normal urine. However, the new rapid and sensitive method clearly revealed its presence. In gastric tissue containing cancer cells, an additional proteinase, which was not present in normal tissue, was detected and named medium moving proteinase (MMP). MMP resembled PGs in alkaline stability rather than the non-PG proteinase, slow moving proteinase (SMP).

Adult↗
PubMed1993

[Human herpesvirus-6 antibodies in idiopathic facial nerve palsy and sudden deafness].

Human herpesvirus-6 (HHV-6) is the causative agent for exanthem subitum. This study investigated the relationship between idiopathic facial nerve palsy (Bell's palsy), sudden deafness and HHV-6 infection. Both Bell's palsy and sudden deafness are syndromes which causes are unknown. Both of them are suspected viral infection as causative agents. Paired sera from 22 patients of Bell's palsy and 39 patients of sudden deafness were examined for reactivity to HHV-6 by the indirect immunofluorescence test. On a case of Bell's palsy and two cases of sudden deafness each of the HHV-6 antibody titers was increased.

Adult↗
PubMed1993

[A stick type filter unit for prevention of biohazard in a research laboratory].

It is important to prevent unexpected laboratory infections in a biosafety area. It is recommended that extreme caution should be used when handling needles and syringes to avoid autoinoculation, and that needles and syringes should be used only when there is no alternative. To remove cells and cell debris, a disk-type microfilter unit, which is recommended using a needle, has been used for filtrating supernatant from HIV-1-infected cell cultures. We compared a stick-type microfilter unit, which can be used without a needle, with a disk-type one for their safety and recovery of human immunodeficiency virus particle after filtration. To confirm the right of our comparison, we observed the filter electron microscopically for the virus particles and virus suspensions were carried out and discussed. In conclusion, the stick-type filter should be improved, since the filter unit is inferior to the disk-type filter unit in purification of the virus, but for the safety is superior to the disk-type.

Acquired Immunodeficiency Syndrome↗
PubMed1992

Immunohistochemical studies on oncogene products (EGF-R, c-erbB-2) and growth factors (EGF, TGF-alpha) in human breast cancer: their relationship to oestrogen receptor status, histological grade, mitotic index and nodal status.

In this investigation, 83 human mammary carcinomas were examined for the expression of oestrogen receptor (ER), epidermal growth factor receptor (EGF-R), epidermal growth factor (EGF), transforming growth factor alpha (TGF-alpha), c-erbB-2, histological grade, mitotic index and nodal status, all of which are reportedly prognostically significant factors (Bloom and Richardson 1957; Baak et al. 1985; Wright et al. 1989). ER expression was biochemically recognized in 43.4% of mammary carcinomas, and EGF-R, EGF, TGF-alpha and c-erbB-2 were histochemically recognized in 25.3, 14.5, 27.7 and 18.0% of mammary carcinomas examined respectively, using conventional sections of buffered formalin-fixed, paraffin-embedded tissue and monoclonal or polyclonal antibodies. There were significant relationships between negative ER and positive EGF-R or TGF-alpha; positive EGF-R and TGF-alpha; positive EGF-R and c-erbB-2; and positive c-erbB-2 and TGF-alpha. The single changes which were the negative ER and the positive c-erbB-2 correlated with histological grade and mitotic index. Co-expression of EGF-R and TGF-alpha correlated with positive nodal status. Therefore, the present investigation indicates that the negative ER, single expression of c-erbB-2 and co-expression of EGF-R and TGF-alpha are important markers which contribute indirectly to prognosis, which reconfirms previous findings on the former two while adding the new finding that immunohistochemical demonstration of expression of EGF-R and TGF-alpha may provide useful information for selecting the appropriate treatment.

Biomarkers, Tumor↗
PubMed1992

A non-radioisotopic reverse transcriptase assay using biotin-11-deoxyuridinetriphosphate on primer-immobilized microtiter plates.

We developed a non-radioisotopic (non-RI) reverse transcriptase assay (RTA). The reverse transcriptase (RT) incorporates biotin-11-deoxyuridine-triphosphate (bio-dUTP) using a poly(rA) template hybridized with oligo(dT) primer that is immobilized on the surface of a 96-well microtiter plate. This assay is thus semi-automated by adapting it to an ELISA testing format. The incorporation of bio-dUTP was enhanced by adding cold dTTP to the reaction mixture, optimally in a molar ratio 4:1 (dTTP:bio-dUTP). This non-RI RTA is more sensitive than the conventional RI assay for the detection of purified Rous-associated virus 2 (RAV-2) and of human immunodeficiency virus type 1 (HIV-1) lysate. Because of its simple procedure, higher sensitivity and non-use of RI materials, the assay can be utilized not only for virological studies but also for routine safety screening of biological products for retroviral contamination.

Avian Leukosis Virus↗
PubMed1992

[An effective concentration method for human immunodeficiency virus type 1 (HIV-1)].

We designed an effective virus concentration method to prevent the infection with human immunodeficiency virus type 1 (HIV-1) in laboratories. The absorbent of Minicon concentrators (Amicon Division, M.R. Grace & Co.-Conn.) was changed to chitin, a mucopolysaccharide extracted from the shells of Japanese pink crab. HIV-1 in the supernatant of HIV-1 infected Molt-4 cells was concentrated by Minicon and the new concentrators. The new concentrator showed good concentration rate and equality of concentration speed.

Chitin↗