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Biomedical subjects

M Horie

Publications and source records attributed to M Horie.

At least 19 recordsLinked to original sources
PubMed1995

Transcriptional induction of pim-1 protein kinase gene expression by interferon gamma and posttranscriptional effects on costimulation with steel factor.

Steel factor (SLF) synergizes with interferon gamma (IFN gamma) to stimulate proliferation of the human factor-dependent cell line MO7e. We examined the effects of IFN gamma and SLF treatment, alone or in combination, on the expression of a 33-kD cytoplasmic protein serine/threonine kinase designated pim-1 whose expression has been closely associated with proliferation induced by related myeloid cytokines. IFN gamma alone, but not SLF, stimulated expression of pim-1 RNA and protein in MO7e cells; compared with IFN gamma alone, costimulation with IFN gamma and SLF resulted in a twofold to threefold increase in pim-1 message and protein expression, correlating with synergistic effects on cell proliferation. Both IFN gamma and IFN gamma/SLF induced pim-1 mRNA in the absence of de novo protein synthesis. Nuclear run-on assays showed that, although IFN gamma alone increased the rate of pim-1 gene transcription, costimulation with IFN gamma and SLF did not further potentiate this effect; however, the stability of pim-1 message was significantly enhanced in the presence of both cytokines. An IFN gamma-responsive element within the 5' flanking region of the pim-1 gene that could confer IFN gamma responsiveness on a heterologous promoter was identified. This sequence, designated PMGAS, formed a specific complex with Stat (signal transducers and activators of transcription) 1 alpha (the p91 subunit of the transcription factor ISGF3 [interferon-stimulated gene factor 3]) in IFN gamma-treated cell extracts, suggesting that the transcriptional effects of IFN gamma on pim-1 expression may be mediated by Stat 1 alpha.

Base Sequence↗
PubMed1995

Cloning and functional characterization of a novel ATP-sensitive potassium channel ubiquitously expressed in rat tissues, including pancreatic islets, pituitary, skeletal muscle, and heart.

ATP-sensitive K+ (KATP) channels play a crucial role in coupling metabolic energy to the membrane potential of cells. We have isolated a cDNA encoding a novel member (uKATP-1) of the inward rectifier K+ channel family from a rat pancreatic islet cDNA library. Rat uKATP-1 is a 424-amino acid residue protein (M(r) = 47,960). Electrophysiological studies of uKATP-1 expressed in Xenopus laevis oocytes show that uKATP-1 is a weak rectifier and is blocked with Ba2+ ions. Single-channel patch clamp study of clonal human kidney epithelial cells (HEK293) transfected with uKATP-1 cDNA reveals that uKATP-1 closes in response to 1 mM ATP and has a single channel conductance of 70 +/- 2 picosiemens (n = 6), indicating that uKATP-1 is an ATP-sensitive inward rectifier K+ channel. In addition, uKATP-1 is activated by the KATP channel opener, diazoxide. RNA blot analysis shows that uKATP-1 mRNA is expressed ubiquitously in rat tissues, including pancreatic islets, pituitary, skeletal muscle, and heart, suggesting that uKATP-1 may play a physiological role as a link between the metabolic state and membrane K+ permeability of cells in almost every normal tissue. Since uKATP-1 shares only 43-46% amino acid identity with members of previously reported inward rectifier K+ channel subfamilies, including ROMK1, IRK1, GIRK1, and cKATP-1, uKATP-1 is not an isoform of these subfamilies and, therefore, represents a new subfamily of the inward rectifier K+ channel family having two transmembrane segments.

ATP-Binding Cassette Transporters↗
PubMed1995

Alloantigen priming induces a state of unresponsiveness in human umbilical cord blood T cells.

Induction of alloreactivity in human adult and umbilical cord blood T cells was evaluated in mixed leukocyte culture by exposure to an allogeneic lymphoblastoid line that expresses known costimulatory molecules. Initial exposure to alloantigen-presenting cells (allo-APC) induced strong proliferative responses in both adult and cord blood T cells. However, in contrast to adult T cells, cord blood T cells exhibited little proliferation after restimulation with donor APC. Primed cord blood T cells could respond to interleukin 2 (IL-2), but unresponsiveness to alloantigen was not overcome by addition of exogenous IL-2. Unresponsiveness was long-lasting and appeared to be maintained by a combination of induction of anergy and activity of CD8+ suppressor cells. This information may contribute to use of human cord blood as an allogeneic source of transplantable stem cells.

Adult↗
PubMed1995

A fast transient outward current in cultured cells from human pulmonary artery smooth muscle.

A voltage-dependent transient outward current (I(to)) was observed in cells cultured from human pulmonary artery smooth muscle when K+, but not Cs+, was the dominant cation in the pipette solution. In 30% of cells investigated using the Cs+ pipette solution, a tetrodotoxin-sensitive inward current (I(in)) dependent on extracellular Na+ was evoked from depolarizations positive to -30 mV.Iin showed voltage-dependent inactivation with a membrane potential at 50% of the evoked current (V50%) of -75.53 +/- 0.81 mV and slope factor potential (Vs) of -10.73 +/- 0.01 mV. In the presence of 1 microM tetrodotoxin, I(to) was rapidly evoked by depolarizations positive to -40 mV and decayed with a single exponential time course (tau = 9.9 +/- 1.1 ms, pulse potential = +50 mV). I(to) also showed voltage-dependent inactivation with a V50% of -70.9 +/- 2.63 mV and Vs of -7.7 +/- 0.03 mV. I(to) was inhibited concentration dependently by the K+ channel blocker, 4-aminopyridine (4-AP), with a concentration of 4-AP at which I(to) was reduced to 50% of control of 36.5 +/- 2.8 microM. These cells possess voltage-dependent currents characteristic of the K(+)-selective fast transient outward and Na(+)-selective inward currents of smooth muscle cells.

4-Aminopyridine↗
PubMed1995

Glibenclamide, an ATP-sensitive K+ channel blocker, inhibits cardiac cAMP-activated Cl- conductance.

Stimulation of the beta-adrenoceptor activates a time-independent Cl- conductance that is known to be regulated via phosphorylation by cAMP-dependent protein kinase in guinea pig ventricular myocytes. Since epithelial cystic fibrosis transmembrane conductance regulator Cl- channels are known to be sensitive to an antidiabetic sulfonylurea, glibenclamide, we tested whether the drug modulates cardiac cAMP-activated Cl- conductance. Bath application of isoproterenol (1 mumol/L, n = 11) or forskolin (1 mumol/L, n = 17) or the intracellular application of cAMP (1 mmol/L, n = 9) activated whole-cell Cl- currents recorded from single myocytes at 36 degrees C. External glibenclamide (> or = 10 mumol/L, n = 26) inhibited the Cl- current induced by either of the stimulants in a concentration-dependent manner. The half-maximal inhibition concentration (IC50) of glibenclamide and the Hill coefficient were 24.5 to 37.9 mumol/L and 1.6 to 2.2, respectively. During current-clamp experiments, forskolin was found to shorten the action potential significantly (250 +/- 45 to 201 +/- 52 milliseconds, P < .05) in 7 of 11 cells tested. Glibenclamide antagonized the forskolin-induced shortening (to 243 +/- 54 milliseconds, n = 7, P < .05). Intracellular administration of sodium orthovanadate (0.5 to approximately 1 mmol/L, n = 6) brought about persistent activation of Cl- current after brief bath application of forskolin. This Cl- current was not affected by H-89 (100 mumol/L, n = 3), a specific inhibitor of cAMP-dependent protein kinase, and was suppressed by glibenclamide similarly, with an IC50 of 29.7 mumol/L.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1995

Lungworm, Filaroides osleri, infection in a dog in Japan.

A 7-month-old male Pomeranian had severe dyspnea for 2 weeks. The lateral bronchogram showed a stenosis of the trachea. Inspite of supportive therapy including supplemental oxygen, the dog died 5 days later. Six pedunculated nodules were recognized in the mucosal surface of the trachea at necropsy. The tracheal nodules were histopathologically granuloma characterized by many coiled parasites containing a little collagen fibers, lymphocytes, plasma cells and macrophages. Female parasites had a lot of embryonated eggs in the uterus. Immature worms were observed in the dilated lymph vessels of bronchial and bronchiolar wall in the lungs. The worms were identified as Filaroides osleri based on the parasitological examinations.

Animals↗
PubMed1995

The combination of Steel factor and GM-CSF blocks apoptosis induced by retinoic acid and upregulates AP-1 in a human growth factor-dependent cell line.

The effects of hematopoietic growth factors were examined on the cellular action of retinoic acid (RA) using the human factor-dependent cell line, MO7e. Treatment of cells with Steel factor (SLF) plus granulocyte-macrophage colony-stimulating factor (GM-CSF) synergistically stimulated cell proliferation compared to that with each factor alone. This synergism was even greater in the presence of RA than in its absence. Treatment of cells with RA resulted in apoptotic cell death associated with internucleosomal DNA fragmentation in the presence of either SLF or GM-CSF. RA-induced apoptosis and DNA fragmentation were completely blocked by treating cells with SLF plus GM-CSF. Northern analysis showed that the inhibition of RA effects on MO7e cells by SLF plus GM-CSF treatment occurred without modulation of expression of RA receptor-alpha (RAR-alpha) gene. Furthermore, a higher amount of AP-1 complex was detected by electrophoretic mobility shift assays in a nuclear extract prepared from cells treated with SLF plus GM-CSF compared to those treated with each factor alone, while the level of RAR-complex remained similar in cells treated with SLF and/or GM-CSF. These data suggest an interaction in signaling pathways among different types of receptors that might be associated with the AP-1 complex.

Apoptosis↗
PubMed1994

Inhibition of the cardiac protein kinase A-dependent chloride conductance by endothelin-1.

Endothelin-1 is a peptide hormone constitutively secreted by vascular and endocardial endothelial cells. Secretion of endothelin-1 is increased under certain pathophysiological conditions, including coronary vasospasm, cardiac ischaemia and myocardial infarction. We have examined the effect of endothelin-1 on the protein kinase A (PKA)-dependent chloride current in voltage-clamped guinea pig ventricular myocytes. This conductance, induced by catecholamines through beta-adrenergic receptors, counteracts the simultaneously increased L-type calcium current by shortening the action potential duration. We report here that endothelin-1, acting through ETA (endothelin-1-selective) receptors, inhibited the current through a pertussis toxin-sensitive mechanism, analogous to muscarinic receptors, by reducing the intracellular cyclic AMP concentration. This effect of endothelin-1 should help protect the ventricle against potentially arrhythmogenic shortening of the action potential during ischaemia when the circulating levels of catecholamines are increased.

Animals↗
PubMed1994

Determination of mirosamicin in animal tissues by high-performance liquid chromatography.

A simple and rapid method using high-performance liquid chromatography (HPLC) for the determination of mirosamicin in animal tissues has been developed. The drug was extracted with 0.3% metaphosphoric acid-methanol (7:3, v/v), and the extracts were cleaned on a Bond Elut SCX (500 mg) cartridge. The HPLC separation was performed on a Puresil 5C18 column (150 x 4.6 mm I.D.) with 0.05 M phosphate buffer (pH 2.5)-acetonitrile (70:30) as the mobile phase at a flow-rate of 0.5 ml/min; the drug was detected at 230 nm with 0.04 AUFS. The calibration graph was linear from 5 to 100 ng. The recoveries of microsamicin from various animal tissues fortified at 1.0 microgram/g were 83.7-88.6% with a relative standard deviation (R.S.D.) of 2.0-5.7%. The detection limit was 0.05 microgram/g.

Animals↗
PubMed1994

Determination of urinary excretion of histamine and 1-methylhistamine by liquid chromatography.

Urinary histamine (Him) and 1-methylhistamine (MH) were determined by liquid chromatography (LC) using on-column derivatization coupled with a column-switching technique. An intact urine sample without any purification was applicable to the LC system because all steps for purification and fluorescence derivatization were fully automated. It was observed that the concentrations of Him and MH increased after hydrolysis of the urine, suggesting the presence of conjugated Him and MH. The level of total/free Him in urine was significantly higher in cancer patients than in normal subjects. Further, a significant correlation between Him and MH was observed in the hydrolysed urine of both normal subjects and cancer patients.

Adolescent↗
PubMed1994

Simultaneous determination of benofloxacin, danofloxacin, enrofloxacin and ofloxacin in chicken tissue by high-performance liquid chromatography.

A simple, rapid and reliable high-performance liquid chromatographic (HPLC) method for the simultaneous determination of residual fluoroquinolones (benofloxacin, danofloxacin, enrofloxacin and ofloxacin) in chicken has been developed. The drugs were extracted with 0.2% metaphosphoric acid-acetonitrile (7:3, v/v), followed by a Bond Elut C18 clean-up procedure. The HPLC separation was carried out on a Wakosil II 5C18-HG column (150 x 4.6 mm I.D.) with 0.05 M phosphate buffer (pH 2.4)-acetonitrile (80:20, v/v) containing 2.5 mM 1-heptanesulfonic acid as the mobile phase. A fluorescence detector was used at an excitation wavelength of 295 nm and an emission wavelength of 455 nm. The calibration graphs were linear from 0.1 to 10 ng for danofloxacin and from 1 to 100 ng for benofloxacin, enrofloxacin and ofloxacin. The recoveries of the drugs from tissues fortified at a level of 0.2 microgram/g were 81.1-89.6%, and the detection limits were 0.01 microgram/g for ofloxacin, danofloxacin and enrofloxacin and 0.02 microgram/kg for benofloxacin. The time needed per sample was less than 60 min.

Animals↗
PubMed1994

Reduced sensitivity of dihydroxyacetone on ATP-sensitive K+ channels of pancreatic beta cells in GK rats.

In the GK (Goto-Kakizaki) rat, a genetic model of non-insulin-dependent diabetes mellitus, glucose-induced insulin secretion is selectively impaired. In addition, it has been suggested by previous studies that impaired glucose metabolism in beta cells of the GK rat results in insufficient closure of ATP-sensitive K+ channels (KATP channels) and a consequent decrease in depolarization, leading to a decreased insulin release. We have recently reported that the site of disturbed glucose metabolism is probably located in the early stages of glycolysis or in the glycerol phosphate shuttle. In the present study, in order to identify the impaired metabolic step in diabetic beta cells, we have investigated insulin secretory capacity by stimulation with dihydroxyacetone (DHA), which is known to be directly converted to DHA-phosphate and to preferentially enter the glycerol phosphate shuttle. In addition, using the patch-clamp technique, we also have studied the sensitivity of DHA on the KATP channels of beta cells in GK rats. The insulin secretion in response to 5 mmol/l DHA with 2.8 mmol/l glucose was impaired, and DHA sensitivity of the KATP channels was reduced in beta cells of GK rats. From these results, we suggest that the intracellular site responsible for impaired glucose metabolism in pancreatic beta cells of GK rats is located in the glycerol phosphate shuttle.

Adenosine Triphosphate↗
PubMed1994

Increased calcium-channel currents of pancreatic beta cells in neonatally streptozocin-induced diabetic rats.

Using a whole-cell patch-clamp technique, voltage-dependent Ca(2+)-channel activities were found to be increased in cultured single beta cells isolated from neonatally streptozocin-induced diabetic rats (NSZ rats). The current-voltage relationship and inactivation time course of Ba2+ currents via L-type Ca2+ channels were indistinguishable between NSZ and control rats. However, the current density observed in NSZ rats was significantly greater than that in control rats. Ba2+ currents via T-type Ca2+ channels were also found to be enhanced in NSZ beta cells. The insulin-secretory capacity of cultured pancreatic islets in response to a depolarizing stimulus (20 mmol/L arginine or 30 mmol/L KCl) in the presence of 11.1 mmol/L glucose was augmented in NSZ rats, whereas that in response to 11.1 and 16.7 mmol/L glucose alone was significantly reduced. It is concluded that the impaired insulinotropic action of glucose in beta cells in NSZ rats is not due to reduced activity of voltage-dependent Ca2+ channels. The fact that insulin secretion induced by a depolarizing stimulus was enhanced in NSZ rats may be related to the augmented activity of the voltage-dependent calcium current found in NSZ beta cells.

Animals↗
PubMed1994

Nitric oxide opens ATP-sensitive K+ channels through suppression of phosphofructokinase activity and inhibits glucose-induced insulin release in pancreatic beta cells.

Nitric oxide (NO) is known to be a potent messenger in the intracellular signal transduction system in many tissues. In pancreatic beta cells, NO has been reported to be formed from L-arginine through NO synthase. To elucidate the effect of NO on insulin secretion and to investigate the intracellular mechanism of its effect, we have used sodium nitroprusside (SNP) as a NO donor. SNP inhibited glucose-induced insulin secretion in a dose-dependent manner, and its effect was reversed by hemoglobin, a known NO scavenger. However, glyceraldehyde-induced insulin secretion was not affected by SNP. Since the closure of ATP-sensitive K+ channels (KATP channel) has been established as a key step in glucose-induced insulin secretion, we have directly assessed the effect of SNP on KATP channel activity using the patch clamp technique. The KATP channel activity reduced by glucose was found to be reversibly activated by the addition of SNP, and this activation was able to be similarly reproduced by applying S-Nitroso-N-acetyl-DL-penicillamine (SNAP), another NO generator. Furthermore, these activating effects were completely eliminated by hemoglobin, in accordance with the reversibility in inhibition of glucose-induced insulin release. However, SNP could not affect the KATP channel suppression by ATP applied to the inside of the plasma membrane. The activation of the KATP channel by NO, therefore, seems to be due to the decreased ATP production attributable to impairment of glucose metabolism in beta cells. Since SNP exhibited no effect on glyceraldehyde-induced KATP channel inhibition, NO may disturb a glycolytic step before glyceraldehyde-3-phosphate. The KATP channel activation by 2-deoxyglucose through presumable ATP consumption due to its phosphorylation by glucokinase was, however, not affected even in the presence of SNP. But in the permeabilized beta cells made by exposure to a low concentration (0.02 U/ml) of streptolysin O (open cell-attached configuration), SNP reopens KATP channels which have been eliminated by fructose-6-phosphate, while this effect was not observed in the KATP channels inhibited by fructose-1,6-bisphosphate. On the other hand, in rat ventricular myocyte KATP channels were not activated by SNP even under a low concentration of glucose. From these observations, the inhibition of phosphofructokinase activity is probably the site responsible for the impairment of glucose metabolism induced by NO in pancreatic beta cells. NO, therefore, seems to be a factor in the deterioration of glucose-induced insulin secretion from pancreatic beta cells through a unique intracellular mechanism.

Adenosine Triphosphate↗
PubMed1994

Extensive proliferative capacity of single isolated CD34 human cord blood cells in suspension culture.

Nonadherent, low-density T-lymphocyte-depleted (NALT-) CD34 cells from normal human cord blood were assessed in suspension culture for the effects of recombinant cytokines on their proliferation, differentiation, and generation of myeloid progenitor cells. In this cell population, 82% of cells expressed c-kit protein as assessed by in situ hybridization, and their cloning efficiency was 85% when cells were plated at low cell numbers with combinations of growth factors. CD34 cells were sorted as 1, 5, or 10 cell(s) per well and also at 5000 cells per dish to initiate stromal-free suspension cultures in the presence of steel factor (SLF), interleukin (IL)-1 alpha, and IL-3. Forty-eight percent of the wells started with a single CD34 cell were positive for growth after 14 days, and the wells contained greater than 5 x 10(3) cells by 21-28 days. Progenitors were assayed weekly with cultures initiated with 1 or 5000 cells. While the fold expansion of nucleated cells was greater in cultures initiated with one cell per well (> 5000 compared to 791-fold expansion for 5000 cells), the fold expansion of progenitors was greater than 5000 cells were used to initiate cultures. Under optimal conditions, there was, respectively, a 160-, 164-, and 57-fold output of high proliferative potential colony-forming cells, granulocyte-macrophage colony-forming units, and erythroid burst-forming units/granulocyte erythroid macrophage megakaryocyte colony-forming units within 1-3 weeks for cultures initiated with 5000 CD34 cells compared with respective fold increases of 29, 16, and 1, for single-initiated cultures. These results demonstrate the expansion capacity of single CD34 cord blood cells and demonstrate that factors in addition to SLF, IL-1 alpha, and IL-3 are necessary for optimal expansion of progenitors from single isolated CD34 cells.

Antigens, CD↗
PubMed1994

Regulation of CFTR channel gating.

Findings outlined here support a complex model for the regulation of cystic fibrosis transmembrane conductance regulator (CFTR) Cl channel gating that incorporates incremental protein kinase A (PKA) phosphorylation of CFTR at multiple sites which, in turn, differentially control the activity of CFTR's two nucleotide-binding domains (NBDs). The NBDs are functionally distinct: only one can respond to the non-hydrolyzable ATP analogue AMP-PNP, and then only after ATP has acted at the other. Moreover, the nature of the responses to AMP-PNP, and to the inorganic phosphate analogue orthovanadate, argues that ATP hydrolysis normally occurs at both NBDs, at one to initiate channel opening and at the other to initiate closing.

Adenosine Triphosphate↗
PubMed1994

Additional similarity of cardiac cAMP-activated Cl- channels to CFTR Cl- channels.

Isoproterenol, forskolin, or cAMP activated a time-independent Cl- current (ICl.cAMP), which is known to be activated by A-kinase-mediated phosphorylation, in guinea pig ventricular myocytes. External glibenclamide inhibited ICl.cAMP in a concentration-dependent manner with IC50 of around 30 microM. Thus, it is concluded that glibenclamide inhibits cardiac cAMP-activated Cl- channels and that the Cl- channel is, in this respect, similar to epithelial CFTR Cl- channel. Northern analysis actually indicated that CFTR messenger RNA is expressed in the guinea pig ventricle.

Animals↗