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F H Ma

Publications and source records attributed to F H Ma.

13 recordsLinked to original sources
PubMed2002

Partial outlet obstruction of the rat bladder induces a stimulatory response on proliferation of the bladder smooth muscle cells.

Outlet obstruction of the rat bladder induces hypertrophy/hyperplasia characterized by increases in bladder mass, smooth muscle content, and collagen deposition. In order to understand the mechanism of the outlet obstruction-induced hypertrophy and hyperplasia, we first determined the temporal pattern of changes in bladder mass after inducing the outlet obstruction. Histological analysis revealed that the smooth muscle cells with hypertrophy and hyperplasia, fibroblasts and connective tissue were increased in a time-dependent manner, corresponding to the temporal pattern observed in the changes in bladder mass, although the phase of changes in these tissue components was somewhat different. In order to further determine whether any proliferation-stimulatory factors were released from the bladder with obstruction in correspondence with increased bladder mass, soluble fractions were prepared from the bladders with outlet obstruction for 3-30 weeks, and their effects on proliferation of smooth muscle cells were examined. The soluble fractions prepared from the bladders at 3 to 14 weeks after obstruction slightly but significantly facilitated the proliferation of cultured smooth muscle cells, while the soluble fractions released after 20 weeks rather suppressed the proliferation. These results suggest that the initial increase in bladder mass might be in part due to the facilitated proliferation of smooth muscle cells of the bladder body induced by growth factors released into the soluble fractions, and that hypertrophy might then play a role in the increased bladder mass at later phases.

Animals↗
PubMed2001

Release of endothelial nitric oxide in coronary arteries by celiprolol, a beta(1)-adrenoceptor antagonist: possible clinical relevance.

Mechanisms underlying celiprolol-induced vasodilatation were analyzed in isolated porcine coronary arteries. Celiprolol induced dose-related relaxation of the artery rings with endothelium, an effect which was suppressed by N(G)-nitro-L-arginine methylester (L-NAME), nitric oxide (NO) scavenger, guanylate cyclase inhibitor, endothelium denudation, and removal of Ca(2+). L-NAME contracted, and superoxide dismutase relaxed, the arteries only when the endothelium was preserved. Neither superoxide dismutase nor beta-adrenoceptor antagonists changed celiprolol-induced relaxations. Celiprolol increased the cyclic GMP content in the tissue. The release of NO from endothelium, estimated by the extracellular production of cyclic GMP in arteries incubated in medium containing guanylate cyclase and GTP, was augmented by celiprolol, and L-NAME abolished this action of celiprolol. It is concluded that celiprolol elicits relaxation by acting on sites other than beta-adrenoceptors in the endothelium and by releasing NO, which activates soluble guanylate cyclase in smooth muscle and produces cyclic GMP. Scavenging of superoxide anions from the endothelium does not seem to account for the induced relaxation.

Adrenergic beta-1 Receptor Antagonists↗
PubMed2001

Identification of muscarinic receptor subtypes of cultured smooth muscle cells and tissue of human bladder body.

BACKGROUND: Muscarinic receptor subtypes of cultured smooth muscle cells from the human bladder body were investigated by the receptor binding assay method. The result was compared with that obtained from the human bladder body tissue to confirm whether the receptor subtypes of the cells are not changed after several passages of cell culture. METHODS: Inhibitory effects of various muscarinic antagonists on the binding of [3H]-N-methylscopolamine ([3H]-NMS) to membrane preparations obtained from cultured smooth muscle cells from the fourth subculture of the human bladder body were compared with those prepared from the human bladder body tissue and cells expressing human muscarinic receptor subtypes. RESULTS: Binding-inhibition constants (pKi) for atropine, pirenzepine, methoctramine, 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP), oxybutynin and propiverine obtained from membrane preparations of cultured smooth muscle cells were 8.91, 6.35, 8.24, 8.53, 7.29 and 5.61, respectively. pKi values of these muscarinic receptor antagonists against the membrane preparation of human bladder body tissue were 9.08, 6.66, 8.05, 8.79, 7.53 and 6.04, respectively. pKi values of cultured smooth muscle cells and tissue from human bladder body were correlated closely with those of insect cells expressing the cloned human M2 receptor subtype. CONCLUSION: The binding affinities for various muscarinic receptor antagonists of cultured human smooth muscle cells were maintained through the fourth subculture and it was suggested that the M2 receptor subtype is predominantly expressed in cultured smooth muscle cells of human bladder body as well as in tissue of the human bladder body.

Cells, Cultured↗
PubMed2000

Determination and bioimaging method for nitric oxide in biological specimens by diaminofluorescein fluorometry.

A simple and sensitive assay and a cellular bioimaging method for nitric oxide (NO) were developed using a novel diaminofluorescein DAF-FM and its diacetate. DAF-FM is converted via an NO-specific mechanism to an intensely fluorescent triazole derivative. For the measurement of NO, the triazole derivative of DAF-FM was determined by reversed-phase high-performance liquid chromatography with fluorescence detection. In the presence of 1 microM DAF-FM, the concentrations of NOR-1, an NO donor, in the range of 2-200 nM were linearly related to the fluorescence intensity. This sensitive NO assay enabled us to detect the spontaneous and substance P-induced NO release from isolated porcine coronary arteries, both of which were dependent entirely on the NO synthase activity in vascular endothelial cells. We also obtained fluorescence images of cultured smooth muscle cells of the rat urinary bladder after loading with DAF-FM diacetate. In the cells pretreated with cytokines, the fluorescence intensity increased with time after DAF-FM loading. This increase in the fluorescence intensity was blocked by prior treatment of the muscle cells with an NO synthase inhibitor, N(G)-nitro-l-arginine methyl ester. Therefore, the present novel diaminofluorescein fluorometry should be useful not only for sensitive NO assay, but also for NO imaging in a variety of biological specimens.

Animals↗
PubMed1998

Inhibition of depolarization-induced nitric oxide synthase activation by NS-7, a phenylpyrimidine derivative, in primary neuronal culture.

Neuronal nitric oxide synthase (NOS) is considered to be involved in the pathogenesis of ischemic brain damage. In the present study, the effect of a novel neuroprotective phenylpyrimidine derivative, 4-(4-fluorophenyl)-2-methyl-6-(5-piperidinopentyloxy)pyrimidine hydrochloride (NS-7), on depolarization-stimulated NOS activity was examined in cultured neurons of mouse cerebral cortex. Various depolarizing stimuli such as veratridine, KCl, and N-methyl-D-aspartate increased the NOS activity determined by cyclic GMP formation. NS-7 concentration-dependently inhibited both the veratridine- and KCl-induced NOS activation with IC50 values of 9.3 and 9.6 microM, respectively. The reversal of KCl-evoked NOS activity by NS-7 was also observed under blockade of both ionotropic glutamate receptors and the Na+ channel with MK-801, 6-cyano-7-nitroquinoxaline-2,3-dione, and tetrodotoxin. In contrast, NS-7, even at 100 microM, did not affect N-methyl-D-aspartate-stimulated NOS activity, nor did it have any influence on NOS activity determined in the soluble fraction of rat hippocampus. Because NS-7 has already been shown to block both Na+ and Ca2+ channels, the present findings suggest that this compound inhibits depolarization-induced NOS activation by reducing Ca2+ influx through blockade of Na+ and Ca2+ channels in primary neuronal culture.

Animals↗
PubMed1997

A novel cognition enhancer NS-105 modulates adenylate cyclase activity through metabotropic glutamate receptors in primary neuronal culture.

The effect of (+)-5-oxo-D-prolinepiperidinamide monohydrate (NS-105), a novel cognition enhancer, on adenylate cyclase activity was investigated in cultured neurons of the mouse cerebral cortex. NS-105 (10(-7) and 10(-6) M) inhibited forskolin-stimulated cyclic AMP formation, an action that was dependent on pertussis toxin-sensitive G proteins. Conversely, in pertussis toxin-pretreated neurons, NS-105 (10(-7)-10(-5) M) significantly enhanced the forskolin-stimulated cyclic AMP formation, and this action was completely reversed by cholera toxin. A metabotropic glutamate receptor agonist (1S, 3R)-1-aminocyclopentane-1,3-dicarboxylic acid (1S, 3R-ACPD) produced similar bi-directional actions on the cyclic AMP formation. Both of these inhibitory and facilitatory actions of NS-105 and 1S, 3R-ACPD were blocked by L(+)-2-amino-3-phosphopropinoic acid (L-AP3). NS-105 (10(-6) M) and 1S, 3R-ACPD (10(-4) M) significantly enhanced isoproterenol- and adenosine-stimulated cyclic AMP formation. The enhancement of such Gs-coupled receptor agonists-stimulated cyclic AMP formation was also produced by quisqualate but not by L(+)-2-amino-4-phosphonobutanoate (L-AP4). The phosphoinositides hydrolysis was enhanced by 1S, 3R-ACPD (10(-4) M) but not by NS-105 (10(-6) M), however, 1S, 3R-ACPD-induced increase in phosphoinositides turnover was attenuated by NS-105. These findings suggest that NS-105 stimulates metabotropic glutamate receptor subclasses that are coupled both negatively and positively to adenylate cyclase, but it acts as an antagonist at the receptor subclasses that are linked to phosphoinositides hydrolysis.

Adenylate Cyclase Toxin↗
PubMed1995

Comparison of fluid volumes with whole bowel irrigation in a simulated overdose of ibuprofen.

OBJECTIVE: To determine the effect of 2 different fluid volumes of polyethylene glycol whole bowel irrigation (WBI) solution on absorption of an ingested toxin in a simulated overdose model. DESIGN: Prospective, randomized, crossover trial. SETTING: Clinical research unit. PARTICIPANTS: Nine adult men. INTERVENTIONS: On 2 separate days, volunteers ingested approximately 75 mg/kg of ibuprofen. In treatment 1, 30 minutes after ingestion of ibuprofen, a 3-L WBI at 2 L/h was begun. This procedure was repeated in treatment 2 with an 8-L. WBI administered at 2 L/h. Fourteen timed serum samples were collected prior to and after drug ingestion for a 24-hour period and analyzed for ibuprofen concentration. The peak serum concentration, time to peak concentration, total area under the serum concentration-time curve (AUC), clearance, and volume of distribution were compared. RESULTS: The mean +/- SD AUCs did not differ between the 3-L (1185.3 +/- 216.9 mg.h/L) and 8-L (1153.5 +/- 251.5 mg.h/L) treatments (p = 0.710). Time to peak serum concentration, peak serum concentration, clearance, and volume of distribution were comparable for the 2 treatments (p > 0.05). CONCLUSIONS: These data indicated that a total WBI volume of 3 L would be expected to perform as well as 8 L administered at the same rate. We recommend that further research define the optimal dose of WBI in acute ingestion of toxins.

Adult↗
PubMed1992

Association of functional alteration in intracellular signal transduction systems with the occurrence of up-regulation of muscarinic receptors in primary cultured neurons.

1. Alterations in intracellular biosignaling systems associated with the up-regulated muscarinic receptor were investigated using primary cultured neurons exposured to 10 nM atropine for 5 days. 2. In neurons treated with atropine, the response of PI turnover to muscarinic stimulation decreased in comparison with that in non-treated neurons. 3. The GTP gamma S-stimulated PI turnover also reduced in neurons possessing up-regulated muscarinic receptor, although phospholipase C activity was not different in these two types of neurons. 4. In addition, the long-term exposure (5 days) to atropine induced the increase in GTPase activity and [3H]GppNHp binding. 5. These results suggest that the decreased response of PI turnover to muscarinic stimulation may be attributed to the functional deterioration of G-protein itself and/or coupling between G-protein and phospholipase C. 6. The present results also strongly suggest that the accentuation of the function of G protein may occur in association with the up-regulation of muscarinic receptor.

Animals↗
PubMed1991

Ontogeny of beta-adrenergic receptor-mediated cyclic AMP generating system in primary cultured neurons.

The developmental changes in the beta-adrenergic receptor/cyclic AMP generating system were examined using mouse cerebral cortical neurons in primary culture. During neuronal growth in vitro, the number of binding sites for [3H]dihydroalprenolol (DHA) showed a tendency to increase (Bmax), while the affinity (Kd) for [3H]DHA did not show any noticeable changes. Basal and isoproterenol-stimulated adenylate cyclase activities as well as the activation of adenylate cyclase by 5'-guanylylimidodiphosphate (GppNHp), NaF and forskolin showed progressive and parallel increases during neuronal growth on a polylysine-coated surface. The treatment of primary cultured neurons with islet-activating protein (IAP), one of the pertussis toxins, attenuated the inhibitory effect of carbachol, a muscarinic agonist, on isoproterenol-induced activation of adenylate cyclase activity. These results indicate that primary cultured neurons possess a cyclic AMP generating system coupled with beta-adrenergic and muscarinic receptors, which is regulated via stimulatory and inhibitory GTP-binding proteins, respectively. The results described above also suggest that the beta-adrenergic receptor, stimulatory and inhibitory types of GTP-binding proteins and adenylate cyclase may develop in a parallel fashion during neuronal growth on a polylysine-coated surface.

Adenylate Cyclase Toxin↗
PubMed1991

Pharmacological characteristics of choline transport system in mouse cerebral cortical neurons in primary culture.

The characteristics of [3H]choline transport with high affinity were investigated using primary cultured neurons obtained from the mouse cerebral cortex. [3H]Choline uptake was saturable as a function of extracellular [3H]choline concentration. Analysis by Lineweaver-Burk plot revealed that [3H]choline was transported into neurons by a high affinity transport system with a Km value of 19.8 +/- 0.8 microM and Vmax value of 0.334 +/- 0.022 nmol/mg protein/min. This high affinity transport of [3H]choline was significantly inhibited by the withdrawal of sodium from the incubation medium, incubation at low temperature (4 degrees C) and addition of metabolic inhibitors such as monoiodoacetate. These results indicate that the high affinity [3H]choline uptake in primary cultured neurons is sodium- and energy-dependent. Hemicholinium-3 also showed a competitive inhibition on the [3H]choline transport. Depolarization by high K+ induced an enhancement of the [3H]choline uptake in the presence of Ca2+. The crude synaptosomal fraction obtained from primary cultured neurons possessed approximately forty-fold higher synthesizing activity of [3H]acetylcholine from [3H]choline than that found in the homogenate preparation of cultured neurons. The present results strongly suggest that the primary cultured neurons used in this study possess a sodium- and energy-dependent high-affinity choline uptake system as well as a synthesizing system for acetylcholine. Possible usefulness of these neurons for investigating neuronal uptake of choline and its functional role in the biosynthesis of acetylcholine are also suggested.

Acetylcholine↗
PubMed1990

Alterations in receptor-coupled second messenger systems at up-regulated muscarinic receptors: analysis using primary cultured neurons.

The effect of a long-term exposure (5 days) to atropine on muscarinic acetylcholine receptors and receptor-coupled second messenger systems was investigated using mouse cerebral cortical neurons in primary culture. The long-term exposure of neurons to atropine (10 nM) induced increases in both the Bmax and Kd values of [3H]quinuclidinyl benzilate (QNB) binding to muscarinic acetylcholine receptors. Alterations in muscarinic receptor-coupled second messenger systems, such as phosphoinositide (PI) hydrolysis and cyclic GMP (cGMP) formation following a long-term exposure to atropine, were also examined. Carbachol-stimulated PI hydrolysis was found to be decreased by the exposure to atropine in spite of the increase of muscarinic receptors. In addition, a long-term exposure to atropine had no effect on carbachol-stimulated cGMP formation as well as on the rightward shift of the carbachol competition curve of [3H]QNB binding in the presence of GTP. These results suggest that the up-regulation in muscarinic cholinergic receptors induced by long-term exposure to atropine may involve not only the increase in number of muscarinic receptors but also the decreased responsiveness in muscarinic receptor-coupled second messenger systems.

Animals↗
PubMedDate not supplied

Low-volume whole bowel irrigation and salicylate absorption: a comparison with ipecac-charcoal.

STUDY OBJECTIVE: To evaluate two methods of gastrointestinal decontamination, low-volume whole bowel irrigation (WBI) and activated charcoal, for their ability to prevent absorption of salicylate. DESIGN: Randomized, two-phase crossover study. SETTING: A clinical research unit in a university-based teaching hospital. PATIENTS: Six healthy, volunteer men. INTERVENTIONS: Subjects were assigned to receive 3000 ml WBI or syrup of ipecac 30 ml followed by activated charcoal 50 g in sorbitol, and were crossed over to the other treatment phase after 1 week. All treatments began 30 minutes after ingestion of 3.25 g aspirin. Urine was collected over 24 hours for analysis of total urinary excretion of salicylate. Serial blood samples were collected for salicylate determination and were subjected to pharmacokinetic analysis. MEASUREMENTS AND MAIN RESULTS: Mean +/- SD recovery of salicylate were WBI 48.6 +/- 5.4% and ipecac-charcoal 37.0 +/- 2.6% from urine (p < 0.01). CONCLUSION: Ipecac-charcoal produced a significantly lower salicylate absorption (peak concentration, AUC) than WBI (p < 0.01) and thus was superior to low-volume WBI.

Adult↗