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Biomedical subjects

N Shimokawa

Publications and source records attributed to N Shimokawa.

11 recordsLinked to original sources
PubMed1992

Characterization of bone protein components with polyacrylamide gel electrophoresis: effects of zinc and hormones in tissue culture.

An attempt was made to clarify the molecular characterization of zinc-induced bone protein synthesis in tissue culture. Calvaria were removed from weanling rat (3-week-old male) and cultured for periods up to 48 hr in Dulbecco's Modified Eagle Medium (high Glucose, 4500 mg/dl) supplemented with antibiotics and bovine serum albumin. When calvaria cultured in the presence of 10(-5) to 10(-4) M zinc were pulsed with [3H] leucine, zinc caused a significant increase in the incorporation of [3H] leucine into the acid-insoluble residues of bone tissue. The soluble fraction obtained from cultured bone was analyzed with SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The major components in the fraction obtained from control bone were 68 killo-dalton (kDa) and 45 kDa proteins. These components were clearly increased by the presence of zinc (10(-4) M). The effect of zinc was completely abolished by the coexistence of 10(-6) M cycloheximide. Meanwhile, 10(-9) M estrogen or 10(-8) M insulin, which can stimulate bone formation, did not enhance the effect of zinc to increase bone 68 and 45 kDa proteins. The present findings suggest that zinc increases many bone protein components, especially 68 and 45 kDa proteins.

Animals↗
PubMed1992

Calcium administration stimulates the expression of calcium-binding protein regucalcin mRNA in rat liver.

The distribution and expression of mRNA encoding the Ca(2+)-binding protein regucalcin in rats were investigated by Northern blot analyses. Liver regucalcin cDNA (0.6 kb) was used as a probe. The analyses of total RNAs extracted from various tissues of rat indicated that regucalcin mRNA was mainly present in liver but only slightly in kidney with a size of 1.8 kb. The expression level decreased with increasing age (3, 10 and 25 weeks). A single intraperitoneal administration of calcium chloride (15 mg Ca/100 g body weight) induced a remarkable increase in regucalcin mRNA in liver; the level was about 200% of control at 30 min after the administration. Subsequently, the expression level began to decrease with time and was about 40% of control level at 120 min after the administration. The increase in regucalcin mRNA levels at 30 min after calcium administration was dose-dependent. These observations show that the expression of regucalcin mRNA is specific in liver of various tissues, and that it is regulated by Ca2+ administration. Regucalcin may have a role as regulatory protein for calcium homeostasis in liver cells.

Animals↗
PubMed1992

Indirect effects of progesterone on the synthesis and secretion of prolactin in mammotroph-enriched cells.

To elucidate the role of steroid hormones in the synthesis and secretion of prolactin (PRL), a mammotroph-enriched cell preparation was made by centrifugal elutriation of trypsin-dispersed anterior pituitary cells from adult male rats. The mammotrophs were enriched by about 2-fold in comparison with those in the initial cell suspension. On the other hand, the ratio of enrichment of gonadotrophs, thyrotrophs and somatotrophs was less than onetenth. The separated mammotrophs were incubated in media containing 17 beta-estradiol and/or progesterone. 17 beta-estradiol increased the levels of PRL in the medium and the cells of 300% and 140%, respectively, and the content of messenger RNA (mRNA) to 193%, whereas progesterone had no such effects. Furthermore, progesterone had no influence on the 17 beta-estradiol-induced increase in the levels of PRL and the contents of PRL mRNA. These results suggest that 17 beta-estradiol regulates PRL synthesis and secretion in mammotrophs, whereas progesterone does not.

Animals↗
PubMed1992

Inhibitory effect of beta-alanyl-L-histidinato zinc on bone resorption in tissue culture.

The inhibitory effect of beta-alanyl-L-histidinato zinc (AHZ) on bone resorption in tissue culture was investigated. Calvaria were removed from weanling rats (3-week-old male) and cultured for periods up to 48 h in Dulbecco's modified Eagle medium (high glucose, 4.5%) supplemented with antibiotics and bovine serum albumin. The experimental cultures contained 10(-7) to 10(-4) mol/l AHZ. The bone-resorbing factors, parathyroid hormone (1-34) (PTH; 10(-7) mol/l), prostaglandin E2 (10(-5) mol/l), interleukin-1 alpha (IL1 alpha; 50 U/ml), and lipopolysaccharide (10 micrograms/ml), caused a significant decrease in bone calcium content. The decreases in bone calcium content induced by bone-resorbing factors were completely inhibited by the coexistence of AHZ (10(-6) to 10(-4) mol/l). Also, AHZ (10(-5) mol/l) completely inhibited the PTH (10(-7) mol/l) or IL1 alpha (50 U/ml)-induced increase in medium glucose consumption and lactic acid production by bone tissue. Furthermore, AHZ (10(-5) mol/l) fairly blocked both PTH (10(-7) mol/l)-increased acid phosphatase and decreased alkaline phosphatase activities of bone tissue. The inhibitory effect of AHZ (10(-5) mol/l) on PTH (10(-7) mol/l)-stimulated bone resorption was clearly prevented by the presence of 10(-4) mol/l dipicolinate, a chelator of zinc. However, zinc sulfate (10(-7) to 10(-4) mol/l) did not inhibit the PTH (10(-7) mol/l)-stimulated bone resorption in tissue culture. These findings indicate that AHZ had a direct inhibitory effect on bone resorption in vitro, and the AHZ effect was found in the chemical form of zinc-chelated dipeptide.

Acid Phosphatase↗
PubMed1991

Oral administration of calcium hydroxide stimulates bone metabolism in the femoral diaphysis of rats with skeletal unloading.

The preventive effect of calcium hydroxide on the disorder of bone metabolism caused by skeletal unloading was investigated. Skeletal unloading was designed using the model of hindlimb hang in rats. Skeletal unloading for 7 d caused a significant decrease of inorganic phosphorus concentration in the serum and of alkaline phosphatase activity and deoxyribonucleic acid (DNA) content in the femoral diaphysis of rats. Oral administration of calcium hydroxide (16 and 24 mg Ca/kg) caused a significant increase in serum inorganic phosphorus concentration and femoral-diaphyseal calcium content and alkaline phosphatase activity of rats with skeletal unloading. Bone DNA content was significantly increased by the dose of 24 mg Ca/kg. These results clearly indicate that skeletal unloading-induced disorder of bone metabolism is partly prevented by oral administration of calcium hydroxide. Calcium ingestion may be useful as a therapeutic tool in the disorder of bone metabolism caused by skeletal unloading.

Animals↗
PubMed1990

Porcine growth hormone: molecular cloning of cDNA and expression in bacterial and mammalian cells.

Porcine growth hormone (PGH) precursor cDNAs were cloned from a pituitary cDNA library constructed in lambda gt11 by immunoscreening. One of the three clones characterized contained an entire nucleotide sequence for the 216-amino-acid precursor molecule. The deduced amino-acid sequence of PGH confirmed the sequence previously reported for that of the genomic DNA of PGH except for one base difference in the coding sequence. Expression of the full-length PGH cDNA was achieved in bacteria and mammalian cells. The mammalian cell line, COS-1, produced the GH molecule which processed the signal peptide and had the same molecular weight as standard PGH, in contrast to the higher molecular weight of the bacterial product. Radioimmunoassay of the recombinant PGH produced in COS-1 cells also revealed an inhibition curve similar to that of the standard PGH.

Amino Acid Sequence↗
PubMed1990

Changes in content of prolactin mRNA during the rat estrous cycle.

An attempt was made to clarify the relationship between the synthesis of prolactin (PRL) and its secretion during the rat estrous cycle by examining the changes in PRL messenger RNA (mRNA); PRL secretion and anterior pituitary PRL content. At various stages of the estrous cycle, pituitary and serum levels of PRL and growth hormone (GH) were measured by radioimmunoassay, while their respective mRNA contents were determined by the hybridization method. The content of pituitary PRL mRNA did not increase in parallel with the increment of the serum level of the hormone. Significant increase in PRL mRNA were observed as three peaks; 1st peak at 14:00 on the proestrus (PE) day, 2nd peak at 4:00 and 3rd peak at 22:00 on the estrus (E) day. The GH levels in the pituitary and serum, or the change in PRL mRNA appeared to be unrelated to the rat estrous cycle. From the present results, we speculate that the three peaks of PRL mRNA content might be ascribable to the direct effect of estrogen on transcription, indirect activation of transcription followed by rapid and massive secretion of PRL, and mammotrope proliferation, respectively.

Animals↗
PubMed1990

Expression of biologically active rat prolactin in mammalian COS-1 cells.

Rat prolactin (PRL) cDNA was constructed in mammalian expression vector, pSVL. Transient expression of rat PRL was performed in COS-1 cells by the DEAE-dextran method. The production of recombinant rat PRL started within 48 h from the cells and reached the level of 1.0-1.5 micrograms/ml/5 x 10(5) cells. The molecular size of recombinant rat PRL was the same as that of standard rat PRL (Mw: 23,000), suggesting successful removal of the signal peptide. The radioimmunoassay and isoelectric focusing analysis showed that recombinant rat PRL has almost the same immunological and biochemical characteristics as those of standard rat PRL. As biological tests, receptor-binding activity, Nb 2 node lymphoma cell growth activity, and mammary gland stimulating activity were examined. The radioreceptor assay showed that recombinant rat PRL has binding activity to mammary microsomal membrane similar to that of standard rat PRL. Recombinant rat PRL also stimulated the growth of Nb 2 lymphoma cells as standard rat PRL. Finally it was shown that recombinant rat PRL promotes the synthesis of the secretory materials in the lumen of mouse mammary gland with the same potency as that of standard rat PRL. In conclusion, recombinant rat PRL, which was produced in mammalian cells in the present experiment, has immunological, biochemical and biological characteristics similar to those of standard PRL, and has full bioactivity.

Animals↗
PubMed1979

Studies on analgesic agents. 1.1a Preparation of 1,2-diphenyl-2-(4-substituted 1-piperazinyl)ethanol derivatives and structure-activity relationships.

The preparation and analgesic activity of a series of the title compounds (8-55 and 57) are described. The intermediates, 2-phenyl-2-(1-piperazinyl)acetophenones 5 and 6, were prepared from benzyl phenyl ketones 3 via their bromides 4. On reduction, compounds 5 afforded the titled compounds 8-12, 16, and 26-48. Compounds 13-15 and 17-25 were obtained by alkylation or benzylation of 1.2-diphenyl-2-(1-piperazinyl)ethanols 7 derived from 6 by reduction. The reduction of 5 and 6 with metal hydrides predominantly gave the erythro isomers. The erythro isomers were remarkably more active than their threo isomers. The more active members in this series of compounds were 16 and derivatives 35 and 37-44 of dl-erythro-1-phenyl-2-(substituted phenyl)-2-[4-(p-methoxybenzyl)-1-piperazinyl]ethanol. Compounds 16, 43, and 44 were the most active with a potency of about two to three times that of codeine. Racemates 16 and 38 were resolved into their optical isomers and it was found that (-)-16 and (+)-38 were more potent than their antipodes. Structure-activity relationship are discussed.

Analgesics↗