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Biomedical subjects

M Kishihara

Publications and source records attributed to M Kishihara.

At least 19 recordsLinked to original sources
PubMed1988

Effect of glucocorticoid administration on intestinal, renal, and cerebellar calbindin-D28K in chicks.

A radioimmunoassay for chick intestinal calcium-binding protein (calbindin-D28K, CaBP-28K) has been developed in our laboratory with a detection limit of 0.3 ng/ml. The values for CaBP-28K in vitamin D-deficient (-D) chicks ranged from a high value for the cerebellum (21,400 +/- 580 ng/mg protein) to a scarcely detectable level in the liver (19.6 +/- 2.2 ng/mg protein). After administration of vitamin D (vitamin D3 500 IU p.o. for 7 days) (+D), the levels of CaBP-28K increased in the duodenum (52,300 +/- 5,100 ng/mg protein), ileum (45,200 +/- 740 ng/mg protein), cerebellum (22,000 +/- 470 ng/mg protein), colon (15,200 +/- 330 ng/mg protein), and kidney (13,460 +/- 540 ng/mg protein). However, the increment in the level of CaBP-28K in each tissue after vitamin D administration was different; levels of CaBP-28K in the duodenum, ileum, and colon increased dramatically more than 200 times after vitamin D administration, whereas that in the kidney showed only a 2.5-fold increase and was unaltered in the cerebellum.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

Identification of parathyroid hormone messenger ribonucleic acid in an apparently nonfunctioning parathyroid carcinoma transformed from a parathyroid carcinoma with hyperparathyroidism.

mRNA coding for pre-pro-PTH, a precursor of PTH, was sought in an apparently nonfunctioning parathyroid carcinoma that had transformed from one that was previously functioning. Total poly(A+) RNA was prepared by phenol-chloroform-isoamyl alcohol extraction and oligo-dT-cellulose affinity chromatography from the tumor tissue and bovine parathyroid glands. In the rabbit reticulocyte lysate cell-free translation system, total poly(A+) RNA from the tumor as well as that from bovine parathyroid glands directed the translation of a product which was specifically precipitated by an anti-PTH serum and which migrated at the same position as pre-pro-PTH on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. These results indicated the presence of mRNA coding for pre-pro-PTH (PTH mRNA) in an apparently nonfunctioning parathyroid carcinoma, suggesting that PTH synthesis is not always absent in parathyroid carcinomas which are not accompanied by hyperparathyroidism.

Animals↗
PubMed1985

Effects of calmodulin antagonists and cytochalasins on proliferation and differentiation of human promyelocytic leukemia cell line HL-60.

To identify the possible roles of Ca2+-related proteins, calmodulin and microfilaments in leukemic cells, we tested the effect of calmodulin antagonists and cytochalasins on proliferation and differentiation of human promyelocytic leukemic HL-60 cells. The growth of HL-60 was inhibited by N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide, N-(4-aminobutyl)-5-chloro-2-naphthalenesulfonamide, and trifluoperazine dihydrochloride. In contrast, the 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]-induced differentiation of HL-60, as judged by plasma-membrane antigenic changes detected by monoclonal antibodies (OKM1, OKT9), nitroblue tetrazolium reduction, and induction of phagocytotic capacity, was not inhibited by N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide or N-(4-aminobutyl)-5-chloro-2-naphthalenesulfonamide, although phagocytosis was depressed by N-(6-amino-hexyl)-5-chloro-1-naphthalenesulfonamide or N-(4-aminobutyl)-5-chloro-2-naphthalenesulfonamide. Trifluoperazine dihydrochloride also failed to inhibit the antigenic change induced by 1,25-(OH)2D3. Cytochalasins B and D, microfilament-disrupting agents, inhibited the cytoplasmic division and the growth of HL-60 but did not inhibit the 1,25-(OH)2D3-induced differentiation. These findings suggest that the calmodulin- and microfilament-dependent process may be involved in the proliferation of HL-60, but not in the differentiation induced by 1,25-(OH)2D3.

Calcitriol↗
PubMed1984

Phenotypic differentiation-linked growth inhibition in human leukemia cells by active vitamin D3 analogues.

Active vitamin D3 induced phenotypic differentiation of the human promyelocytic leukemia cell line, HL-60, cultured in serum-free medium. Upon exposure to 10(-10)-10(-7) M 1,25-(OH)2D3, 1,24S-(OH)2D3 or 1,24R(OH)2D3, monocyte-granulocyte-associated plasma membrane antigens of HL-60 cells detected by monoclonal antibodies, OKMI, 63D3 and Mo2, quantitated by fluorescence-activated cell sorter analysis, were increased time- and dose-dependently. After expose to 1,25-(OH)2D3, promotion of this antigenic expression was detected within 16 h, and the induction of differentiation continued until 96 h. The number of cells bearing transferrin receptors recognized by the monoclonal antibody, OKT9, and its density on the surface of HL-60 cells were decreased symmetrically. These effects appeared in parallel with the inhibition of cell growth, poly(ADP-ribose) content and de novo DNA-RNA synthesis. These findings indicate that 1,24S-(OH)2D3 stimulates differentiating induction of HL-60 like 1,25-(OH)2D3 in vitro and that the decrease of transferrin receptor is apparently correlated with the inhibition of proliferation.

Antibodies, Monoclonal↗
PubMed1984

Low molecular weight thyroglobulin leading to a goiter in a 12-year-old girl.

We characterized the abnormal thyroglobulin (TG) in the thyroid and serum of a 12-yr-old girl with a large sporadic multinodular goiter first noted at age 4 yr. She developed normally and had no clinical evidence of hypothyroidism. However, her serum T4 was less than 1.0 microgram/dl, T3 was 125 ng/dl, and TSH was 155 microU/ml. Serum PBI was 9.7 micrograms/dl, and more than 90% was not extractable with butanol. The 24-h radioactive iodine uptake was 55%, not dischargeable by perchlorate. Hormone formation was tested by the administration of 131I before surgery. [131I]T4 and [131I]T3, but not 131I-labeled iodotyrosines, were present in the thyroidal venous blood. Hydrolysis of 10,000 X g supernatants from three randomly obtained samples of the goiter revealed 66-77% of the 131I as iodotyrosines, 2-4% as iodothyronines, and 10-12% as undigestable material; the MIT to DIT ratio ranged from 3.1-8.7, and the T4 to T3 ratio ranged from 2.3-8.3. The TG level was 2.5 mg/g in the goiter and 9.4 micrograms/ml in the serum. The RIA displacement curves for the goiter and serum TG levels were both identical to the curve produced by normal human TG. The iodine contents of goiter and serum TG were 0.49% and 0.47% (wt/wt), respectively. The T4 to T3 ratio was lower in the goiter (approximately 5) than in the serum iodoprotein (approximately 45), whereas the calculation of the T4 to T3 ratio in the thyroidal secretion was less than 1. The goiter and serum TG bound normally to Concanavalin A, indicating that they contained carbohydrate. When either serum- or goiter-soluble proteins were gel-filtered (Bio-Gel A-5m), TG immunoreactivity and stable iodine elution profiles were the same, suggesting that no significant amounts of other iodoproteins were present in the thyroid or circulation. Both serum and goiter TG elution volumes corresponded to mol wt of approximately 9 X 10(4). A sedimentation rate of 10-11 S was found for both goiter and serum TG. An abnormally low mol wt of 8.5-9.0 X 10(4) was determined by sodium dodecyl sulfate-electrophoresis, in good agreement with the estimates from gel filtration studies. A single band was present on sodium dodecyl sulfate-electrophoresis regardless of whether the TG was reduced before the analysis. Thus, it is very unlikely that the low molecular weight was due to partial hydrolysis.(ABSTRACT TRUNCATED AT 400 WORDS)

Child↗
PubMed1982

Paradoxical anti-leukemic effects of plant-derived tumor promoters on a human thymic lymphoblast cell line.

Three different diterpen polyols and the teleocidin have antileukemic effects on the human thymic leukemia cell line HPB-ALL by inducing phenotypic differentiation. We tested TPA (phorbol esters), mezerein (daphnane), milliamin (ingenol ester) and teleocidin B (teleocidins) as representative of the chemical structure of each principle and found that these agents have the similar biological activity of inducing phenotypic differentiation in HPB-ALL cells. Moreover, competitive binding of [3H]PDB to cell-surface receptors was significantly inhibited by these agents; n-butyrate and phorbol per se did not inhibit the binding of [3H]PDB. Despite differences in the structure of the principles, these agents may have similarly acting side-chain structures.

Alkaloids↗
PubMed1982

Effect of various drugs on the binding of thyrotrophin to thyroid plasma membranes.

Effects of enzyme inhibitors and membrane-active drugs on the binding of 125I-labelled thyroid-stimulating hormone (TSH) to human thyroid membranes and membrane adenylate cyclase (AC) activity were studied. FOY, a synthetic polyvalent proteolytic enzyme inhibitor, Trasylol, alpha- and beta-adrenergic blocking agents, tranquilizers, anti-histamines and polyene antibiotics enchanced TSH binding in a dose-dependent manner, whereas selective enzyme inhibitors and adrenergic stimulating agents had no effect. Both propranolol and FOY inhibited basal and TSH stimulated AC activity of thyroid membranes. FOY, as well as propranolol was found to have protective effects on hypotonic erythrocyte lysis. These results suggest that propranolol and FOY increased TSH binding by the same mechanism, probably the so-called membrane-stabilizing effects. Although the detailed mechanisms underlying the increased TSH binding by these drugs remain unknown, they may change the membrane structure, thereby enhancing the TSH receptor affinity.

Adenylyl Cyclases↗
PubMed1981

Differences between circulating and tissue thyroglobulin in rats.

The mechanism whereby thyroglobulin (TG) reaches the circulation can involve either the release of newly synthesized TG or the release of colloid-stored TG from the thyroid gland. To distinguish between these possibilities, we have compared the properties of circulating and glandular TG in normal and thyroidectomized thyroid tumor-bearing rats. Circulating TG had the properties of poorly iodinated molecules; it was more susceptible to dissociation into subunits and had a lower density, the latter determined by equilibrium centrifugation in concentrated RbCl. The density of circulating TG was the same as that of glandular TG from propylthiouracil-treated rats, suggesting that circulating TG was nearly or completely devoid of iodine. Circulating TG bound to Concanavalin A-Sepharose and had a normal MCR, indicating that mannose was present and galactose was not in terminal positions, both properties of glandular TG. Since previous studies suggest that these properties cannot arise from differential clearance of TG molecules in the periphery, these data suggest that the TG in the circulation may arise from the direct release of poorly iodinated newly synthesized TG from the thyroid.

Animals↗
PubMed1981

Interaction between thyrotropin (TSH) binding inhibitor immunoglobulins (TBII) and soluble TSH receptors in fat cells.

To clarify whether TSH binding inhibitor immunoglobulins (TBII) are antibodies to the membrane site associated with the TSH receptor itself or its neighboring sites, the interactions of TSH and TBII with soluble TSH receptor were investigated with a TSH radioreceptor assay using labeled highly purified bovine TSH (bTSH) and Triton extracts from guinea pig crude fat cell membranes (800-10,000 x g fraction). Treatment of the crude fat cell membranes with 0.5% (vol/vol) Triton X-100 resulted in solubilization of membrane proteins with recoveries of 25-30%, whereas increasing the concentration of Triton X-100 in the assay medium caused a decrease of [125I]bTSH binding to the solubilized membranes. Thus, the solubilized membranes were found to retain the capacity to bind [125I]bTSH below the Triton X-100 concentration of 0.4% in the assay medium. Incubation of the solubilized membranes with [125I]bTSH for 24 h at 4 C led to a steady state of specific binding, while incubation at 37 C resulted in more rapid but less specific binding, with a shorter duration of the steady state. Maximum binding occurred within the physiological pH range. Both TSH and Graves' immunoglobulins (Igs) specifically inhibited [125I]bTSH binding to the solubilized membranes, as demonstrated by polyethylene glycol separation of the [125I]bTSH-solubilized membrane complex from unbound [125I]-bTSH. Scatchard analysis of [125I]bTSh displacement curves for both TSH and Graves' Igs indicated a single class of binding site for each, with an affinity constant of 1.8 x 10(9) M-1. In addition, Igs capable of inhibiting [125I]bTSH binding to the solubilized membranes were detected predominantly in the serum of patients with Graves' disease. These results strongly suggest that TBII are antibodies directed to the TSh receptor itself without strict organ and species specificity.

Adipose Tissue↗
PubMed1981

Genetic and clinical studies of serum beta 2-microglobulin levels in haematological malignancies.

Sera from 244 patients with haematological malignancies were examined for beta 2-microglobulin (beta 2m) levels. There were 142 leukaemias, 32 malignant lymphomas, three immunoblastic lymphomas, two pseudolymphomas and 65 multiple myelomas. Culture supernatants from various established cell lines were also tested. The phenotype facilitating beta 2m shedding from the cell surface appeared to be independent of the specific IgG heavy chain allotypes; however, a myeloma group with normal serum beta 2m levels showed a significant association with the specific Gm allotypes. The determination of serum beta 2m levels can provide valuable information on the proliferative stage of the disorders, the effectiveness of chemotherapy, and be a diagnostic aid for blastic crisis in chronic myelocytic leukaemias, and for subtyping lymphoid malignancies.

Beta-Globulins↗
PubMed1980

Immunologic studies on Werner's syndrome.

Five patients with Werner's syndrome were studied to elucidate the relationship between the immune system and the ageing process. Three of five patients had neoplasms such as fibrosarcoma, meningioma and thyroid adenoma but none of the five patients manifested symptoms and signs suggestive of typical autoimmune disorders. Most immunohaematological data were normal; however, two patients exhibited diminished lymphocyte functions such as a significant (P less than 0 . 05) reduction in pokeweed mitogen (PWM) induced B cell differentiation, a slightly decreased response to Con A- or mitomycin-treated allogeneic lymphocytes and naturally occurring serum antibody to T cells. While these immunological changes were quantitatively small, subnormal levels of immune functions were detected. Cultured fibroblasts exhibited a decrease in clonal growth potentials and elongation of newly synthesized DNA, a decreased expression of HLA antigens, and a diminished secretion of beta 2-microglobulin.

Adult↗
PubMed1979

Interaction between thyroid-stimulating immunoglobulins and thyrotropin receptors in fat cell membranes.

In the TSH radioreceptor assay to study the interaction between Graves' immunoglobulins (Ig) and TSH receptors in guinea pig fat cell membranes, Graves' Ig were found to inhibit [125I]TSH binding to fat cell membranes in a dose-dependent manner. Scatchard analysis of [125I]TSH displacement curves by Graves' Ig indicated a single population of the binding sites in fat cell membranes, in contrast to two populations of TSH-binding sites in the membranes. Displacement of [125I]TSH bound to fat cell membranes by both Graves' Ig and unlabeled TSH were time and temperature dependent, with similar dissociation curves, suggesting a specific binding of Graves' Ig to the membrane sites related to the TSH receptor in the fat cells. Such Ig are referred to as fat cell-binding Ig, to be distinguished from the thyroid-stimulating Ig (TSI) detected by TSH radioreceptor assay using human thyroid membranes. Both fat cell-binding Ig and TSI were detected in the sera of a great majority of untreated patients with Graves' disease. A significant correlation was found between both values (r = 0.80; n = 19; P less than 0.001). According to these results, TSI might represent an autoantibody to the membranes associated with the TSH receptor of the target tissues without a strict organ specificity.

Adipose Tissue↗