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Biomedical subjects

T Koda

Publications and source records attributed to T Koda.

At least 55 records · Page 3Linked to original sources
PubMed1995

Pyrolysis-gas chromatography of carbonate apatites used for sintering.

Gas chromatography was employed to quasi-continuously determine the amount of carbon dioxide that evolved from carbonate apatite specimens during sintering. Assuming that the carbonate in the specimens decomposed to carbon dioxide on a mole-for-mole basis, the determination of the carbon dioxide evolved allowed for the determination of the amount of carbonate that remained in the specimens during different stages of sintering. Previously, this measurement could be carried out only after sintering was completed. Comparison of data obtained from specimens compacted isostatically at 600 MPa for sintering with powder specimens indicated that the amount of carbonate remaining in the sintered apatite mass strongly depended on heating rates, heating temperatures, and holding-time intervals.

Apatites↗
PubMed1995

Comparison of the type-2 insulin-like growth factor receptor in normal osteoblasts and osteosarcoma-derived osteoblast-like cells.

Insulin-like growth factor-II is known to stimulate the proliferation and differentiation of osteoblasts in part through activation of the type-2 insulin-like growth factor receptor. The present study examined the type-2 insulin-like growth factor receptors of three normal osteoblast-like cells and three osteosarcoma-derived osteoblast-like cells (OGA, SU, and IMAI) from humans. [125I]insulin-like growth factor-II was used for the binding studies. All of the cell types had high affinity binding sites for insulin-like growth factor-II (dissociation constants [Kd] < or = 1 nM). The concentration of these sites was 10 to 24-fold higher in normal osteoblasts than in the osteosarcoma cells studied. Unlabeled insulin-like growth factor-II inhibited the binding of [125I]insulin-like growth factor-II to the cells in a dose-dependent manner; however, unlabeled insulin-like growth factor-I and insulin were less effective. Covalent crosslinking of insulin-like growth factor-II binding sites gave molecular mass estimates of M(r) 250,000 in human osteoblast cells, 250,000 and 130,000 in OGA cells, 240,000 in SU cells, and 250,000 and 130,000 in IMAI cells. Unlabeled insulin-like growth factor-II inhibited all affinity labeling. In Northern blot analysis, the type-2 insulin-like growth factor receptor mRNA of normal osteoblasts was seen in greater abundance than it was in osteosarcoma cells. These results indicate that the numbers of type-2 insulin-like growth factor receptors differ between normal and transformed osteoblasts and that the differential expression of the receptor may be due to the differentiation of osteoblasts.

Adolescent↗
PubMed1995

17 beta-estradiol increases the receptor number and modulates the action of 1,25-dihydroxyvitamin D3 in human osteosarcoma-derived osteoblast-like cells.

It is well known that 17 beta-estradiol (E2) and 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) have important roles in bone metabolism. This study was undertaken to examine E2 regulation of 1,25(OH)2D3 receptor (VDR) expression and the biological action of 1,25(OH)2D3 in human osteoblast-like cells. When human osteosarcoma-derived osteoblast-like cells were treated with varying concentrations of E2, the VDR levels increased by up to 100% in a dose-dependent manner. VDR levels significantly increased at 10 nM E2 and this increase plateaued at 100 nM E2. E2-dependent increase of VDR was time dependent, plateauing at 24 hours and was maintained for at least 48 hours in human osteoblast-like cells. Scatchard analysis showed that E2 increased the number of VDR (12.3 +/- 0.4 versus 26.5 +/- 0.3 fmol/mg protein; mean +/- SE of three independent experiments) rather than the Kd (0.15 +/- 0.02 versus 0.16 +/- 0.01 nM; mean +/- SE of three independent experiments). Tamoxifen (50 nM), a specific competitor with E2, completely abolished the E2-induced increase of VDR. The levels of VDR mRNA (4.5 kb) from the cells increased in a dose-dependent manner after E2 treatment. With regard to the biological effects, E2 increased by 10-25% the inhibitory effect of 1,25(OH)2D3 on cell growth. However, E2 did not increase the stimulation of alkaline phosphatase activity by 1,25(OH)2D3. The present study suggests that E2 modulates the biological action of 1,25(OH)2D3 through VDR levels in bone cells.

Alkaline Phosphatase↗
PubMed1994

An upstream NF-Y-binding site is required for transcriptional activation from the hst promoter in F9 embryonal carcinoma cells.

Expression of hst (k-FGF, FGF-4), a member of the fibroblast growth factor gene family, is restricted to early stages of developing embryos and to embryonal carcinoma cells. In F9, which is a prototype of embryonal carcinoma cells expressing hst, the expression of hst gene is positively regulated by a downstream octamer motif that functions as an enhancer. We have investigated, by chloramphenicol acetyltransferase (CAT) reporter fusion gene analysis in F9, the cis-acting regulatory element within the hst promoter region that interacts with this enhancer. Electrophoretic mobility shift assay and methylation interference analysis showed that the hst promoter contains, in a segment termed Y, the sequence 5'-CTGATTGGCA-3', which closely resembles the consensus binding motif for the CCAAT-binding factor NF-Y. Deletions or mutations in this element substantially reduced expression of hst-CAT constructs. The nuclear factor binding to the Y segment of the hst promoter was indistinguishable from NF-Y, as inferred from interactions with specific anti-NF-Y monoclonal and polyclonal antibodies. We conclude that the expression of the hst gene in F9 is positively regulated by the coordinated interaction between an NF-Y-binding site and an octamer motif.

Animals↗
PubMed1994

Regulatory sequences required for hst-1 expression in embryonal carcinoma cells.

The hst-1 gene, which is implicated in mammalian embryonic development and morphological transformation of NIH3T3 cells, is expressed in undifferentiated F9 cells, but not in differentiated F9 and other well-differentiated cells, such as PYS-2, NIH3T3 and HeLa cells. An octamer element present in the 3' untranslated region acts as an enhancer. Although Oct3 is down-regulated when F9 cells are differentiated, transient expression of Oct3 did not enhance the hst-1 promoter activity in HeLa, NIH3T3 or PYS-2 cells. Thus, the role of Oct3 on hst-1 expression remains elusive, and an additional transcription factor which interacts may regulate hst-1 transcription in association with Oct1, Oct3 or both.

3T3 Cells↗
PubMed1994

Hepatitis C virus genotypes, reactivity to recombinant immunoblot assay 2 antigens and liver disease.

To clarify the relationship between hepatitis C virus (HCV) genotypes and liver disease, we typed HCV genomes in the sera of 151 blood donors, 180 patients with type C chronic liver disease (CLD), and 30 haemophiliacs residing in Hiroshima, Japan. All of the subjects were positive for anti-HCV and HCV-RNA, and were examined for seroreactivity to HCV-specific antigens. The HCV genotypes were determined by polymerase chain reaction (PCR) with type-specific primers deduced from the putative core region of the HCV genome. Significantly more (P < 0.001) type III HCV was found in the samples from the CLD patients (80%) than in those from the blood donors (55%). Significantly more (P < 0.001) type III HCV was found in the samples from the blood donors (29.1%) than in those from the CLD patients (11.7%). There was no significant difference in the distribution of the HCV types among the patients with chronic active hepatitis, liver cirrhosis, and hepatocellular carcinoma. A four-antigen recombinant immunoblot assay (RIBA-2) assay was used to compare the serum samples for their reactivity to a range of structural and nonstructural peptides specific for HCV (5-1-1, C100-3, C33c, and C22-3). The frequency of seropositivity to 5-1-1 and C100-3 was significantly higher (P < 0.001) in type II HCV-infected blood donors than in type III HCV-infected donors (68.2% and 65.9% vs. 4.5% and 22.7%, respectively). Among the type III HCV-infected individuals, the CLD patients had a significantly higher (P < 0.01) frequency of seropositivity to 5-1-1 than the blood donors (33.3% vs. 4.5%).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

[Diagnosis of hepatocellular carcinoma using chicken anti-monoclonal antibody--as tumor marker on the heterophilic hanganutziu-deicher antigen].

The Hanganutziu-Deicher (HD) antigen is a heterophilic antigen with N-glycolylneuraminic acid as the terminal carbohydrate, and has attracted much attention as one of tumor-associated antigens. This antigen is absent from normal tissues of human and chicken but expressed in human neoplasms. Extensive research on HD antigen has not be made yet to show the intimate correlation with human neoplasm, but it is attracting an attention as a diagnostic marker for the definition of cancer. We made two different chicken monoclonal antibodies against HD antigen as a pathological tool for hepatocellular carcinoma (HCC). The hybrid cell, which produced antibodies, were prepared by means of fusion technology with HD3 antigen-immunized chicken spleen cells and HAT-sensitive chicken B cell line termed R27H4. The two antibodies had a different characteristic, one recognized only glycolipid, and the other recognized glycoprotein as well as glycolipid. Glycolipid type HD antigen and glycoprotein type HD antigen were identified by using two chicken anti-HD monoclonal antibodies. Glycolipid type HD antigen was detected by flow cytometry in 4 out of 24 HCC samples (87.5%). In our study, increased serum levels of IgG type and/or IgM type HD antibodies were observed in 21 out of 24 patients with HCC (87.5%). The presence of these antibodies is attributable to the expression of the HD antigen in cells of HCC. Eleven out of 24 patients with HCC were negative for AFP and PIVKA-II, and 10 out of these 11 patients (90.9%) were positive for anti-HD antibodies. We consider that patients with HCC should be screened for HD antigen and antibody for an early diagnosis of hepatocellular carcinoma; the same as for AFP and PIVKA-II both of which are tumor-associated antigens that have been associated with for hepatocellular carcinoma.

Animals↗
PubMed1993

Molecular cloning of a cDNA encoding a novel small GTP-binding protein.

A cDNA encoding a small GTP-binding protein, S10, was cloned from Jurkat cells. The deduced amino acid sequence of S10 had the structural features characteristic to this family of proteins with highest homology to rab subfamily. Northern blot analysis revealed that this gene is expressed only in lymphoid cell lines and a histiocytic leukemia, U937. Hence, it should have a specialized function in cells derived from the hematopoietic stem cell.

Amino Acid Sequence↗
PubMed1993

Heterogeneity of HLA-G genes identified by polymerase chain reaction/single strand conformational polymorphism (PCR/SSCP).

A genomic HLA-G clone named 7.0E was isolated from a Japanese placenta. The deduced amino acid sequence of the 7.0E was identical to two HLA-G genomic clones and two cDNA clones previously described. The DNA sequences of alpha 1 and alpha 2 domains of the HLA-G gene from 5 cell lines also encoded the same amino acids. However, a 14 bp insertion, ATTTGTTCATGCCT, was present in the 3' untranslated region of 7.0E compared with the originally described HLA-G clone (HLA 6.0). Polymerase chain reaction (PCR)/single strand conformational polymorphism (SSCP) analysis of exon 8 allowed the HLA-G gene to be classified into two alternative types, G6.0 and 7.0 E, those correlated to the absence or the presence of the 14 bp stretch. Each group had minor sequence variant(s), and the alleles of the 7.0E-type were more heterogeneous than those of the G6.0-type. The 14 bp deletion is present only in the G6.0-type of HLA-G alleles among HLA class I genes. Thus it was suggested that G6.0 alleles were generated after diversification of the HLA-G.

Alleles↗