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T Kono

Publications and source records attributed to T Kono.

At least 253 records · Page 14Linked to original sources
PubMed1990

The human interleukin-2 receptor beta-chain gene: genomic organization, promoter analysis and chromosomal assignment.

The chromosomal gene for the human interleukin-2 receptor beta-chain (IL-2R beta) was isolated and characterized. The entire IL-2R beta gene is composed of ten exons spanning about 24.3 kilobases, in which the protein is encoded by the exons 2-10. The cysteine rich extracellular region which displays a significant evolutionary resemblance to other cytokine receptors, as well as growth hormone and prolactin receptors, is encoded primarily by exons 3 and 4, whereas the membrane proximal, cysteine poor domain showing a homology with type III modules of fibronectin is encoded by exon 7. Sequence analysis of the 5'-flanking region revealed the presence of potential binding sites for transcription factors such as Octamer binding factors, AP-1, AP-2 as well as the 'GC-clusters'. At least five potential cap sites were identified by S1 mapping analysis. The 850 bp DNA sequence of the 5'-flanking region exhibited constitutive promoter activity when it was linked upstream of the HSV-tk reporter gene and then transfected into YT cells, a human leukemic cell line. By applying the RFLP linkage analysis, the IL-2R beta gene has been assigned to chromosome 22q12-13.

Base Sequence↗
PubMed1990

Eclosion hormone of the silkworm Bombyx mori. Expression in Escherichia coli and location of disulfide bonds.

A gene encoding eclosion hormone (EH) from the silkworm, Bombyx mori was chemically synthesized, inserted into a secretion vector and expressed in Escherichia coli, leading to the production of biologically active EH. Sequence analysis of cystine-containing peptides in a thermolysin digest of this EH established the locations of 3 disulfide bonds in the molecule. Evidence was also obtained that the 6 residues at the NH2-terminal are dispensable but 4 residues at the COOH-terminal play an important role in EH activity.

Amino Acid Sequence↗
PubMed1990

Stimulation of the insulin-sensitive cAMP phosphodiesterase by an ATP-dependent soluble factor from insulin-treated rat adipocytes.

The insulin-sensitive cAMP phosphodiesterase (PDE) in the microsomal fraction (Fraction P-2) from basal (-insulin) rat adipocytes was stimulated upon incubation with 2 mM ATP plus the soluble fraction from insulin-treated adipocytes (Fraction S-2+). Fraction S-2+ was prepared in the presence of p-nitrophenylphosphate, sodium vanadate, and EGTA. The ATP-dependent stimulation of PDE was routinely 60-70%. The unknown factor in Fraction S-2 was water-soluble, heat-labile, excluded by Sephadex G-50, mostly retained by Sephadex G-100, and not inhibited with 1 microgram/ml heparin, 3 mM CaCl2, or 30 mM NaF. The soluble factor may be a mediator of insulin action on PDE, possibly a protein kinase.

3',5'-Cyclic-AMP Phosphodiesterases↗
PubMed1990

Cell cycle analysis of human dermal fibroblasts cultured on or in hydrated type I collagen lattices.

The proliferation and cell cycle phase composition of human dermal fibroblasts cultured on or in type I collagen lattices (reconstituted dermis model) were examined. On collagen lattices, as compared with conventional cultures on plastic dishes, the proliferation of human dermal fibroblasts was suppressed, being arrested at about one-half the saturation density after 10 days of culture. In collagen lattices, proliferation was further suppressed, being nearly arrested within 4-7 days of culture. Cells were analyzed for cell cycle phases by two-color flow cytometry using DNA staining and S phase cell staining with FITC-conjugated antibromodeoxyuridine antibody. After 5 days of culture, the number of S phase cells on collagen lattices was 49.3% of that on plastic dishes, with an increase in G0G1 phase cells of 79.8%. In collagen lattices, the number of S phase cells was very small (4.3% of all cells), and most of the cells accumulated in G0G1 phase. These findings suggest that the cell cycle of fibroblasts is arrested at G0G1 phase by their interaction with collagen. On the basis of these results, the reconstituted dermis model using collagen lattice is considered to be analogous to the dermis in vivo with respect to cell growth and cell cycle phase composition.

Cell Cycle↗
PubMed1990

Enhanced melanogenesis of murine melanoma cells cultured on or in collagen gel.

To elucidate the interaction between melanoma and its matrix, we cultured B16 murine melanoma cells on and in type I collagen gel and evaluated specified functions of melanoma cells; tyrosinase activity and melanin-synthesizing capacity. Proliferation of cells cultured in these environments was markedly suppressed compared with that of cells cultured conventionally on plastic. On the other hand, the tyrosinase activity of cells cultured in or on collagen gel was two to three times higher than that of cells cultured on the plastics, while their melanin production was approximately double that achieved during conventional culture of cells. In conclusion, collagen gel influenced the growth and cell-specific functions of the melanoma cell. The culture system using collagen gel as substrate may be useful for the investigation of the interaction between melanoma and its matrix.

Animals↗
PubMed1990

Murine interleukin 2 receptor beta chain: dysregulated gene expression in lymphoma line EL-4 caused by a promoter insertion.

The functional, high-affinity interleukin 2 receptor (IL-2R) consists of at least two receptor components, IL-2R alpha (p55) and IL-2R beta (p70-75). The cDNA encoding the murine IL-2R beta has been isolated by using the previously cloned cDNA for human IL-2R beta as a probe. Analysis of the cDNA revealed that the murine IL-2R beta shows a marked homology with the human IL-2R beta and that it is also structurally related to other cytokine receptors such as erythropoietin receptor. The cDNA-directed murine IL-2R beta formed high-affinity IL-2R in conjunction with the endogenous IL-2R alpha in a murine pro-B-cell line and could transduce IL-2-induced growth signal. In mouse lymphoma line EL-4, the IL-2R beta gene was found to be rearranged by the insertion of the long terminal repeat sequence of an intracisternal A particle, giving rise to constitutive expression of the IL-2R beta mRNA.

Amino Acid Sequence↗
PubMed1990

Parallel arrangement, growth inhibition and cell cycle phase analysis of human dermal fibroblasts cultured in collagen lattice.

Human dermal fibroblasts were cultured in a hydrated type I collagen lattice. When collagen fibers were arranged in one direction, fibroblasts were arranged in the same direction. Cell proliferation was markedly suppressed in the collagen lattice as compared with that on plastic, with growth being arrested after day 5. No differences in proliferation were observed between aligned cells and randomly oriented cells. Flow cytometry with DNA staining was performed to analyze each phase of the cell cycle of fibroblasts. Among the 10,000 cell population, S phase cells on day 2 of culture accounted for 43% on plastic but were markedly inhibited to 25% in the lattice. On day 4, S phase cells accounted for 33% on plastic but only for 10% in the lattice. These findings suggest that cell advancement to the S phase is markedly inhibited in the collagen lattice, resulting in accumulation of most of cells in the G0G1 phase. The present study clearly showed that culture in the collagen lattice allowed alignment of fibroblasts with a definite orientation as observed in vivo and produced a status resembling that in vivo in terms of proliferation and cell cycle phase composition.

Adult↗
PubMed1990

Correlation of contractility and proliferative potential with the extent of differentiation in mouse fibroblastic cell lines cultured in collagen lattices.

Four types of fibroblastic cell lines at various stage of differentiation, which had been derived from syngeneic mice, were cultured in collagen lattices (reconstituted dermis model). Lattice contraction, growth in the lattice, and cell morphology were compared. The following cell lines were used: [I] precrisis cells within several subcultures derived from the skin of Balb/c mice, [II] an established normal cell line derived from syngeneic mice (Balb/3T3 clone A31), and [III] two transformed lines (Balb/3T12-3, 3T3-B-SV40) originating from [II]. The cells adopted a bipolar spindle form in the collagen lattice. Lattice contraction was the most marked with cell type [I] followed in order by [II] and [III]. Relative growth in the lattice occurred in the reverse order (III greater than II greater than I). These findings suggested a correlation between lattice contraction and growth in the lattice and also between the extent of differentiation and lattice contraction.

Animals↗
PubMed1990

Proliferative potential of murine melanoma cells cultured in or on collagen gel.

As the first step in developing an in vitro model of melanoma cells infiltrating the dermis, B16 murine melanoma cells were cultured on and in type I collagen gels. Under these conditions, the melanoma cell adopted an elongated or dendritic form. Cell proliferation was suppressed in the culture system using the collagen gel as compared with the conventional monolayer culture on plastic. Microcinematographically, this suppression was found to be due to an extension of the cell cycle time of each individual cell. On the other hand, there were no appreciable differences in proliferation pattern between the cells cultured on type I and IV collagen film and those cultured on plastic. These results suggest that there are interactions between type I collagen in the gel form and melanoma cells, especially with respect to cell growth.

Animals↗
PubMed1990

Effects of human recombinant tumor necrosis factor-alpha (TNF-alpha) on the proliferative potential of human keratinocytes cultured in serum-free medium.

The effects of human recombinant tumor necrosis factor-alpha (TNF-alpha) on human keratinocytes cultured in a serum-free medium were investigated. TNF-alpha markedly suppressed cell growth. The growth-inhibitory effect was reversible and cytostatic at a concentration of 1-5 U/ml, but appeared to be irreversible and cytocidal at 10 U/ml. The growth suppressive effect was more marked when TNF-alpha was added in the late growth phase or preconfluent phase than when it was added in early or mid-growth phases. No effects of TNF-alpha on cell adhesion to the substrate were observed. These results indicate that TNF-alpha is a very potent anti-proliferative agent for human keratinocytes.

Cell Adhesion↗
PubMed1990

Infiltration of melanoma cells into the type I collagen gel.

Melanoma cells were cultured on type I collagen gel, and the infiltration of of those cells into the gel was observed. B16 murine melanoma cells initially adopted a spherical form on the gel, but they assumed a dendritic form after infiltration into the interior. The degree of infiltration increased very rapidly and was time-dependent. No correlations between the growth rate or melanogenic activity and infiltrative potential were observed. When Syrian hamster and human melanoma cell lines were cultured, the degrees of infiltration varied. This culture system using collagen gel is considered to be a useful in vitro model of tumor cell invasion.

Animals↗
PubMed1990

PUVA increases ornithine decarboxylase gene expression in mouse skin.

The influence of topical PUVA on the activity of ornithine decarboxylase (ODC) and its gene expression was investigated in the skin of hairless mouse. After 1 h of application of 0.3% 8-methoxypsoralen, irradiation of 3 J/cm2 of ultraviolet A (UV-A) was administered. The ODC activity markedly increased and peaked at 24 h following UV-A irradiation. The ODC mRNA level, analyzed in dot-blot analysis, elevated to about 5 times that of control at 24 h after treatment. These results show that the PUVA-induced ODC activity is due in part to an increase in ODC gene expression.

Animals↗
PubMed1990

Contraction phenomenon of type I collagen gel by melanoma cells.

In order to study the interaction between melanoma cells and collagen, B16 murine melanoma cells were embedded and cultured in type I collagen gel. Melanoma cells cultured in the collagen gel became elongated, as compared with those cultured on plastic, and some of them assumed a dendritic form. The gel contracted very slowly but steadily during culturing of melanoma cells, as in the experiment using fibroblasts. On the 20th day of culture the area of the gel accounted for only 32% of that when culture started. This contraction was enhanced by retinoic acid, which is known to induce cell differentiation. The contractility of the gel differed between various lines of melanoma cells. The present observations raise the possibility of interaction between melanoma cells and type I collagen.

Animals↗
PubMed1990

Correlation between ageing and collagen gel contractility of human fibroblasts.

To investigate the influence of ageing on wound healing, we cultured fibroblasts derived from human dermis in type I collagen gel, and evaluated the relationship between gel contractility and ageing. Cells were obtained from children (0-15 years old, Group A), early adulthood (16-40 years old, Group B), mid-adulthood (41-60 years old, Group C), and the elderly (61 or older, Group D). Gel contractility was determined by measuring the diameter on the second day after gel preparation. Within the tenth passage, gel contraction was the most marked in Group A, but did not differ among the other groups. Gel contraction at passages 30-40 did not differ from those within the tenth passage in Groups B, C and D, but it decreased markedly in Group A to a value similar to that in the other groups. These results show that fibroblasts in childhood are more contractile than those in adulthood and are more readily affected by passages (in vitro ageing).

Adolescent↗
PubMed1990

Possible formation of cutaneous amyloid from degenerative collagen fibers. Ultrastructural collagen changes and the immunoreactivity of cutaneous amyloidosis employing anti-type I, III, IV, V collagen antibodies.

The origins of primary cutaneous amyloid have been investigated and several possibilities are proposed. We investigated here several cases by electron microscopy and found in all cases characteristic filamentous changes which were identical with amyloid structures in the collagen bundles. These observations suggested a possible pathway of cutaneous amyloid formation from degenerative collagen fibers. Further studies were performed using immunoelectron microscopical and immunohistochemical methods employing a panel of anti-collagen antibodies to examine the reactivity of amyloid to them. Our results were much more suggestive of the collagen origin of this kind of amyloid.

Amyloid↗
PubMed1990

[Experimental studies of retinal glial cell proliferation on retinal surface].

Our previous reports described the formation processes of epiretinal membranes composed of retinal glial cells in the experimental vitreous hemorrhage. This report presents the pathological changes of retinal glial cells, inner limiting membranes and vitreous body in the early stage of experimental vitreous hemorrhage. Using an operating microscope, 0.3 ml of autologous whole blood was injected through the pars plana into the vitreous cavity of albino rabbits. The eyes were enucleated at 3, 7, 10, 14 and 28 days after injection. Three days after injection, normal vitreous structure disappeared and vitreous fibers condensed. Seven days after injection, red blood cells were found on the retinal surface. The inner limiting membrane, composed of lamina rara and lamina densa, had a normal appearance. Where the red blood cell were present on the retinal surface, the lamina rara of the inner limiting membrane became thin. Fourteen days after injection, where glial cells showed upheaval, lamina rara disappeared and lamina densa became thin. Afterward, glial cells extruded their cytoplasmic processes onto the retinal surface. Twenty-eight days after injection, the inner limiting membranes were normal in appearance except for the area of glial cells penetrating the inner limiting membranes. The results suggested that, in vitreous hemorrhage, glial cell proliferation onto the retinal surface is an active reaction in order to engulf the debris of the red blood cells.

Animals↗