Search PubMed⌕ Search

Biomedical subjects

M Okuno

Publications and source records attributed to M Okuno.

At least 109 records · Page 6Linked to original sources
PubMed1995

Role of SPan-1 antigen in adhesion of human colon cancer cells to vascular endothelium.

Recently E-selectin (ELAM-1, endothelial leukocyte adhesion molecule-1) was shown to recognize not only sialyl Lewis X but also sialyl Lewis A, and these carbohydrate antigens may be involved in the process of the adhesion between cancer cells and endothelial cells in cancer metastasis. To investigate the contribution of sialylated carbohydrate antigen, SPan-1, and sialic acid to the adhesion of human colon cancer cells to endothelial cells, adhesion assay using HUVECs (human umbilical vein endothelial cells) was performed. The adhesion was significantly inhibited by pretreatment with anti-E-selectin antibody, indicating that this adhesion was thought to be mediated by E-selectin. When these cancer cells were pretreated with SPan-1 antibody, the adhesion was significantly inhibited in a concentration-dependent manner. The adhesion was also inhibited by pretreatment with neuraminidase. These findings suggest that the SPan-1 antigen plays a significant role in the adhesion of human colon cancer cells to endothelial cells, and sialylation of the terminal structure of carbohydrate antigens is important in this adhesion.

Antigens, Neoplasm↗
PubMed1995

Stimulatory effect of angiotensin II on calcium efflux from cultured bovine adrenal chromaffin cells.

The effect of angiotensin II on Ca2+ mobilization in cultured bovine adrenal chromaffin cells was examined. Angiotensin II (10(-7)M) increased the intracellular free Ca2+ level ([Ca2+]i) to a peak in the presence or absence of extracellular Ca2+, followed by decrease with time. Angiotensin II (10(-9)-10(-6)M) also stimulated 45Ca2+ efflux from cultured bovine adrenal chromaffin cells in a concentration-dependent manner. Its stimulatory effect on 45Ca2+ efflux was inhibited by the angiotensin II antagonist [Sar1, Ile8]-angiotensin II or [Sar1, Val5, Ala8]-angiotensin II. The increase in angiotensin II-stimulated 45Ca2+ efflux was dependent on the extracellular Na+ concentration. Angiotensin II also increased 22Na+ influx into the cells. These results indicate that stimulation of the angiotensin II receptor induces extracellular Na(+)-dependent Ca2+ efflux from cultured bovine adrenal chromaffin cells, probably by acceleration of Na+/Ca2+ exchange.

Adrenal Medulla↗
PubMed1995

Calcium efflux from cultured bovine adrenal chromaffin cells induced by pituitary adenylate cyclase-activating polypeptide (PACAP): possible involvement of an Na+/Ca2+ exchange mechanism.

The effect of pituitary adenylate cyclase-activating polypeptide 1-38 (PACAP1-38) on Ca2+ efflux from cultured bovine adrenal chromaffin cells was examined. PACAP1-38 stimulated the efflux of 45Ca2+ from the cells in a concentration dependent manner (10(-9)-10(-7)M). This effect was inhibited by its potent receptor antagonist PACAP6-38. PACAP1-38 increased the formation of [3H]inositol phosphates and cyclic AMP in the cells. Forskolin, an activator of adenylate cyclase, also stimulated the efflux of 45Ca2+ from the cells. 3-Isobutyl-1-methylxanthine (IBMX), an inhibitor of phosphodiesterase, enhanced PACAP1-38-induced 45Ca2+ efflux from the cells. Phorbol 12-myristate 13-acetate (PMA), an activator of protein kinase C, had no effect on the efflux of 45Ca2+ from the cells. The increases in 45Ca2+ efflux induced by PACAP1-38 and forskolin were reduced by deprivation of extracellular Na+ and the Na+/Ca2+ exchange inhibitor amiloride. In addition, PACAP1-38 stimulated 22Na+ influx into the cells, and this action was inhibited by amiloride. These results suggest that PACAP1-38 stimulates an Na+/Ca2+ exchange mechanism through activation of adenylate cyclase in cultured bovine adrenal chromaffin cells.

1-Methyl-3-isobutylxanthine↗
PubMed1995

Lysophosphatidic acid as a stimulator of Na(+)-dependent Ca2+ efflux from adrenal chromaffin cells.

The effect of lysophosphatidic acid (LPA) on Ca2+ efflux from cultured bovine adrenal chromaffin cells was examined. LPA stimulated the efflux of 45Ca2+ from the cells in a concentration dependent manner (10(-7)-10(-5)M). Other lysophospholipids (lysophosphatidylcholine, lysophosphatidylethanolamine, lysophosphatidylinositol and lysophosphatidylserine), phosphatidic acid, monoolein and oleic acid had not significant effect on 45Ca2+ efflux from the cells. The LPA-stimulated 45Ca2+ efflux was dependent on the extracellular Na+ concentration and was inhibited by amiloride, inhibitor of Na+/Ca2+ exchange. LPA increased 22Na+ uptake into the cells and this effect was also sensitive to amiloride. These results indicate that LPA stimulates extracellular Na(+)-dependent 45Ca2+ efflux from cultured bovine adrenal chromaffin cells, probably through its stimulatory effect on membrane Na+/Ca2+ exchange.

Animals↗
PubMed1995

Regulations by cyclic nucleotides and phorbol ester of calcium efflux from cultured bovine adrenal chromaffin cells.

The effects of cyclic nucleotides and phorbol ester on Ca2+ efflux from cultured bovine adrenal chromaffin cells were examined. Dibutyryl cyclic AMP (DB-cAMP), forskolin (an activator of adenylate cyclase), dibutyryl cyclic GMP (DB-cGMP) and nitroprusside (an activator of guanylate cyclase) all stimulated 45Ca2+ efflux from the cells preloaded with 45Ca2+. These agents did not increase the intracellular free Ca2+ ([Ca2+]i) level. On the contrary, phorbol 12-myristate 13-acetate (PMA; an activator of protein kinase C) did not affect the efflux of 45Ca2+, but inhibited the increase in 45Ca2+ efflux caused by DB-cAMP, forskolin, DB-cGMP or nitroprusside. The 45Ca2+ effluxes stimulated by cyclic nucleotides, forskolin and nitroprusside were inhibited by deprivation of extracellular Na+ ([Na+]o). These results suggest that both cAMP- and cGMP-dependent protein kinases are involved in the stimulatory mechanism of [Na+]o dependent Ca2+ efflux, probably through acceleration of [Na+]o/[Ca2+]i exchange and that protein kinase C plays an inhibitory role in this mechanism.

Animals↗
PubMed1995

D2 receptors in the ventrolateral striatum are involved in feeding behavior in rats.

To study the role of dopamine D1 and D2 receptors in the ventrolateral striatum in feeding behavior, a D1 receptor agonist (CY 208-243), a D1 receptor antagonist (SCH 23390), a D2 receptor agonist (quinpirole), and a D2 receptor antagonist [(-)-sulpiride] were perfused via a microdialysis probe into the ventrolateral striatum of rats fasted for 22 h. Then the rats were allowed to feed freely for 6 h. Sulpiride perfusion at a high concentration suppressed food and water intake significantly, whereas dopamine release and the levels of DOPAC and HVA were increased at all concentrations. In contrast, quinpirole perfusion at a high concentration increased food intake by 41%. Dopamine release and the levels of DOPAC and HVA were decreased at all concentrations. On the other hand, neither CY 208-243 nor SCH 23390 changed food intake or dopamine release, but both drugs decreased water intake. These results suggest that D2 receptors in the ventrolateral striatum have a more important role than D1 receptors in the feeding behavior of rats.

3,4-Dihydroxyphenylacetic Acid↗
PubMed1995

Activation of mammalian sperm motility by regulation of microtubule sliding via cyclic adenosine 5'-monophosphate-dependent phosphorylation.

Bicarbonate was found to be essential for activating live mouse sperm motility. The activated sperm flagella exhibited high beat frequency, high swimming velocity, and large principal and reverse bends. To gain further insight into the bicarbonate-triggered activation mechanism, the microtubule sliding characteristics of the activated versus the nonactivated sperm flagella were compared by use of demembranated sperm. We found that the effects of bicarbonate on live sperm were identical with the effects of cAMP on demembranated sperm both in microtubule sliding velocity and in sliding disintegration pattern. Furthermore, autoradiography revealed that the activation of mouse sperm motility was associated with cAMP-dependent phosphorylation of a 65-kDa flagellar protein. The results demonstrated that bicarbonate-triggered activation of mouse sperm motility was closely coupled with the regulation of microtubule sliding via cAMP-dependent phosphorylation.

Adenosine Triphosphate↗
PubMed1995

Pituitary adenylate cyclase-activating polypeptide stimulates the synthesis of dopamine in cultured bovine adrenal chromaffin cells.

The effect of pituitary adenylate cyclase-activating polypeptide1-38 (PACAP1-38) on the synthesis of dopamine in cultured bovine adrenal chromaffin cells was examined. PACAP1-38 stimulated [14C]dopamine synthesis from [14C]tyrosine, in a concentration-dependent manner, causing maximal stimulation at 10(-7)M. This stimulatory action of PACAP1-38 was not significantly inhibited by staurosporine (an inhibitor of protein kinase C) or in the cells in which protein kinase C was down-regulated by prolonged exposure to TPA (an activator of protein kinase C), whereas it was partially attenuated in Ca(2+)-free medium. PACAP1-38 increased the formation of [3H] inositol phosphates, [Ca2+]i, 45Ca2+ uptake and cAMP level. The peptide also stimulated the phosphorylation of tyrosine hydroxylase, the enzyme catalyzing the rate-limiting step in dopamine synthesis. Dopamine synthesis and tyrosine hydroxylase phosphorylation stimulated by the maximal effective concentration of dibutyryl cAMP or high K+, which activates Ca2+ uptake, were further enhanced by PACAP1-38. These results indicated that PACAP1-38 may stimulate the activities of cAMP- and calcium-dependent protein kinases in cultured bovine adrenal chromaffin cells, resulting in increase in the synthesis of dopamine probably by stimulation of phosphorylation of tyrosine hydroxylase.

Animals↗
PubMed1995

Effects of branched-chain amino acid infusion on protein metabolism in rats with acute hepatic failure.

Branched-chain amino acids (BCAA) are known to improve hepatic encephalopathy as well as protein malnutrition in cirrhosis. However, such effects in acute hepatic failure (AHF) remain to be elucidated. The current study was conducted to investigate whether BCAA improves protein metabolism in AHF. AHF was induced in male Donryu rats weighing approximately 230 g by giving 60 mg/kg lipopolysaccaride intravenously and 800 mg/kg D-galactosamine hydrochloride intraperitoneally. From 18 hours after injection, AHF rats and control rats were given one of the following five solutions intravenously for 6 hours: 1) saline, 2) 10% glucose, 3) standard 10% amino acid formula with total nitrogen content of 12.2 g/L and BCAA/aromatic amino acid molar ratio of 37.05, 4) BCAA-enriched solution with nitrogen content of 21.9 g/L and the ratio of 148.2, or 5) an active placebo against BCAA-enriched solution with nitrogen content of 21.9 g/L and the ratio of 37.05. In parallel, each group was given a continuous infusion of 14C-leucine. After the plasma radioactivity of 14C-leucine and the expired 14CO2 level reached a plateau, protein turnover was analyzed according to the kinetic model proposed previously by Waterlow. When compared with the control, rates of total protein turnover (total flux), oxidation, and breakdown all increased significantly in AHF. Infusion of standard 10% amino acid formula, BCAA-enriched solution or the placebo in AHF increased total flux and oxidation significantly as compared with the effect of saline or 10% glucose. Although saline, 10% glucose, standard 10% amino acid formula, and the placebo had no effect on synthesis rate, it was increased significantly with BCAA-enriched solution.(ABSTRACT TRUNCATED AT 250 WORDS)

Acute Disease↗
PubMed1995

[The changes in nuclear DNA content in acquisition of a high capacity for metastasis of human cell lines].

Quantification of nuclear DNA content in cancer is considered a prognostic factor, and several investigators have demonstrated that aneuploid tumors are more aggressive than diploid tumors. In this report, we investigated the relationship between metastatic capacity and nuclear DNA content using gastric and colon cancer cell lines. We established a highly peritoneal metastatic cell line (2M-D3) from a gastric cancer cell line (2M) and a highly liver metastatic cell line (LM-H3) from colon cancer cell line (LM) in our laboratory. Both the highly metastatic cell lines have more capacity for growth and metastasis, but lower nuclear DNA content and chromosome numbers than these parental cell lines. The results indicated that metastatic capacity was inversely proportional to nuclear DNA content. We cannot provide a satisfactory explanation for this inverse correlation between metastatic capacity and nuclear DNA content, but it is supposed that each cancer cell may have suitable and stable nuclear DNA content in metastasis.

Animals↗
PubMed1995

Regulation of adipocyte gene expression by retinoic acid and hormones: effects on the gene encoding cellular retinol-binding protein.

Our laboratory has reported that adipose tissue and adipocytes are importantly involved in retinoid storage and metabolism. To gain further insight, we examined factors that may regulate CRBP mRNA levels in primary cultures of rat epididymal and murine BFC-1 beta adipocytes. Northern blot analysis revealed that retinoic acid is a potent inducer of CRBP mRNA, causing a 7.5-fold rise in mRNA levels in primary adipocytes and a 9.5-fold rise in BFC-1 beta adipocytes. This induction of CRBP mRNA was dose-dependent at retinoic acid concentrations ranging between 10(-8) and 10(-5) M. Retinoic acid induction of CRBP mRNA levels showed a short lag period (6 h) and reached a maximal level of induction by 12 h in BFC-1 beta adipocytes and by 48 h in primary epididymal adipocytes. Nuclear run-on transcription assays of retinoic acid-induced BFC-1 beta adipocytes indicated that the rate of CRBP gene transcription is enhanced 3.6- to 4.3-fold by retinoic acid. In contrast, dexamethasone markedly down-regulated CRBP expression in a dose-dependent manner at concentrations ranging between 10(-9) and 10(-6) M. CRBP mRNA levels in primary and BFC-1 beta adipocytes declined, respectively, by 90% and 80% when adipocytes were exposed to 10(-6) M dexamethasone for 24 h. Studies of mRNA half-life indicated that dexamethasone acts to lessen CRBP expression through the specific destabilization of CRBP mRNA. Treatment of both primary and BFC-1 beta adipocytes with triiodothyronine alone had no effect on CRBP mRNA levels; however, when adipocytes were treated with a mixture of triiodothyronine and retinoic acid, the induction of CRBP mRNA levels by retinoic acid was reduced. In summary, these studies indicate that CRBP gene expression is regulated by retinoic acid, dexamethasone, and triiodothyronine; thus suggesting that retinol uptake, intracellular transport, and metabolism are dynamically regulated in adipocytes.

Adipocytes↗
PubMed1995

Increased expression of sialyl Lewis A and sialyl Lewis X in liver metastases of human colorectal carcinoma.

Sialyl Lewis A (SLA) and sialyl Lewis X (SLX) have been shown to be specific ligands for endothelial leukocyte adhesion molecule-1 (ELAM-1), and may be involved in the process of adhesion between cancer cells and endothelium. We used immunohistochemical methods to study the expression of SLA, SLX and CEA in both primary tumors and matched metastatic liver lesions of colorectal carcinomas. Specimens from primary tumors and matched liver metastases from 24 patients with colorectal carcinomas were studied immunohistochemically. The degree of expression of CEA in liver metastases was similar to that in primary tumors, but SLA and SLX were expressed on a larger proportion of tumor cells in liver metastases than in primary tumors. Our findings suggest that colorectal carcinoma cells expressing SLA and/or SLX form metastatic liver tumors. They also suggest that expression of SLA and SLX in primary of colorectal carcinoma can be used as a prognostic indicator of metastasis.

Adult↗
PubMed1995

[Carotenoids].

Explore the source record for details and available documents.

Adult↗
PubMed1994

Mechanism of histamine-induced calcium efflux from cultured bovine adrenal chromaffin cells: possible involvement of an Na+/Ca2+ exchange mechanism.

The effect of stimulation of the histamine receptor on Ca2+ mobilization in cultured bovine adrenal chromaffin cells was examined. Histamine (10(-5) M) increased the intracellular free Ca2+ ([Ca2+]i) to a peak in the presence or absence of extracellular Ca2+, followed by decrease with time. Histamine (10(-8)-10(-5) M) also stimulated 45Ca2+ efflux from cultured bovine adrenal chromaffin cells in a concentration dependent manner. Its stimulatory effect on 45Ca2+ efflux was inhibited by the specific histamine H1 receptor antagonist mepyramine. The increase in histamine-stimulated 45Ca2+ efflux was inhibited by deprivation of extracellular Na+ and by the Na+/Ca2+ exchange inhibitor amiloride. In addition, histamine stimulated 22Na+ influx into the cells, and this action was inhibited by amiloride. These results suggest that stimulation of the histamine H1 receptor regulates Na+/Ca2+ exchange in cultured bovine adrenal chromaffin cells.

Adrenal Medulla↗
PubMed1994

A new cancer-associated antigen defined by a monoclonal antibody against a synthetic carbohydrate chain.

Carbohydrate antigens can be designed by referring to previously defined carbohydrate structures. We have generated a novel monoclonal antibody (MAb) (F1 alpha-75) against an artificially designed antigen (F1 alpha), using organic-synthetic chemistry methods and hybridoma technology. F1 alpha (Gal beta 1-->4GlcNAc beta 1-->6GalNAc alpha 1-->Ser/Thr) belongs to core type 6 of O-linked glycans, which has not been previously reported in human cancers. To produce antibodies against F1 alpha, a glycolipid was synthesized which carries the carbohydrate portion of F1 alpha on a ceramide foundation (Gal beta 1-->4GlcNAc beta 1-->6GalNAc alpha 1-->Cer). The MAbs we obtained (F1 alpha-75, F1 alpha-87) specifically recognized F1 alpha and had only a very weak or no cross-reactivity with other glycolipids similar to F1 alpha. We investigated the expression of F1 alpha in human tissues, including 110 gastric cancers, 73 colon cancers and 42 pancreatic cancers. F1 alpha was found in human cancerous tissues but not in normal adult tissues. The rate of positive staining with F1 alpha-75 was 80.0% for gastric cancer, 52.4% for pancreatic cancer and 38.4% for colon cancer. F1 alpha-75 also reacted with the tissues neighboring gastric and pancreatic tumors but not intensely. Among fetal tissues, F1 alpha-75 reacted with the pyloric glands of the stomach, the centro-acinar cells of the pancreas, the convoluted tubules of the kidney and the terminal bronchioles of the lung.

Antibodies, Monoclonal↗
PubMed1994

Behavior of Japanese tree frogs under microgravity on MIR and in parabolic flight.

Japanese tree frogs (Hyla japonica) were flown to the space station MIR and spent eight days in orbit during December, 1990. Under microgravity, their postures and behaviors were observed and recorded. On the MIR, floating frogs stretched four legs out, bent their bodies backward and expanded their abdomens. Frogs on a surface often bent their neck backward and walked backwards. This behavior was observed on parabolic flights and resembles the retching behavior of sick frogs on land--a possible indicator of motion sickness. Observations on MIR were carried out twice to investigate the frog's adaptation to space. The frequency of failure in landing after a jump decreased in the second observation period. After the frogs returned to earth, readaptation processes were observed. The frogs behaved normally as early as 2.5 hours after landing.

Adaptation, Physiological↗
PubMed1994

Alterations of O-glycan biosynthesis in human colon cancer tissues.

Human colon cancer is associated with antigenic and structural changes in mucin-type carbohydrate chains (O-glycans). To elucidate the control of the biosynthesis of these O-glycans is colon cancer, we have studied glycosyltransferase and sulphotransferase activities involved in the assembly of elongated O-glycan structures. We analysed homogenates prepared from cancer tissue, adjacent normal and distal normal tissue from 20 patients. Several transferase activities showed pronounced changes in cancer tissue. The changes correlate with previous findings of a loss of O-glycans in cancer mucins, but did not always correlate with levels of Tn, sialyl-Tn, T and Lex antigens in homogenates or with the differentiation status and Duke's stages of the cancer tissue or the patient's blood type, sex and age. UDP-GlcNAc: Gal NAc-R beta 3-N-acetylglucosaminyltransferase (where GlcNAc is N-acetyl-D-glucosamine and GalNAc is N-acetyl-D-galactosamine) synthesizing O-glycan core 3, GlcNAc beta 1-3GalNAc-, CMP-sialic acid: GalNAc-peptide alpha 6-sialyltransferase synthesizing the sialyl-Tn antigen and sulphotransferase activities towards O-glycan core 1, Gal beta 1-3GalNAc-, were found to be decreased in cancer. UDP-GlcNAc: Gal beta 1-3GalNAc beta 6-N-acetylglucosaminyltransferase was also decreased in cancer concomitant with a loss of the ability to synthesize the I antigen and core 4, GlcNAc beta 1-6(GlcNAc beta 1-3) GalNAc-, CMP-sialic acid: Gal beta 1-3GalNAc-R alpha 3-sialyltransferase and GDP-fucose: Gal beta-R alpha 2-fucosyltransferase, synthesizing the blood group H determinant, were found to be 4- and 3- to 8-fold increased, respectively, in cancer compared to normal tissue. The data suggest that the biosynthesis of antigens and mucin-bound O-glycan structures in colon cancer is subject to complex control mechanisms.

Adult↗