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Biomedical subjects

H Kanaide

Publications and source records attributed to H Kanaide.

At least 145 records · Page 8Linked to original sources
PubMed1989

Increased fibrinopeptide A during anginal attacks in patients with variant angina.

It is not known whether coronary vasospasm is associated with coronary thrombosis. In this study, plasma levels of fibrinopeptide A during anginal attacks in 24 patients with variant angina were examined. A hyperventilation test was used to induce angina. Hyperventilation induced angina and ST segment elevation (AST: 0.32 +/- 0.14 mV, p less than 0.01) in eight patients with variant angina. Fibrinopeptide A increased from 0.75 +/- 0.27 at control to 7.8 +/- 4.4 ng/ml (p less than 0.01) during anginal attacks in these eight patients. In addition, four patients had spontaneous attacks of angina; they also had elevated levels of fibrinopeptide A during attacks (from 2.0 +/- 1.2 at control to 21.9 +/- 18.0 ng/ml [p less than 0.01] during attacks). Hyperventilation did not induce either angina or ST segment elevation in 12 of the patients with variant angina. Fibrinopeptide A levels did not change with hyperventilation in these patients. To determine whether elevated plasma levels of fibrinopeptide A were associated with angina, the plasma levels of fibrinopeptide A were examined during exercise-induced angina in seven additional patients with stable effort angina. They all developed angina with treadmill exercise; however, plasma fibrinopeptide A did not change. Therefore, only the patients with variant angina demonstrated elevated levels of fibrinopeptide A during anginal attacks. These findings suggest that coronary vasospasm associated with myocardial ischemia may induce stasis of blood, resulting in fibrinogen-fibrin conversion in the coronary vessels.

Angina Pectoris, Variant↗
PubMed1989

Effects of okadaic acid on cytosolic calcium concentrations and on contractions of the porcine coronary artery.

1. We investigated the effects of okadaic acid (OA), a phosphatase inhibitor derived from a 38-carbon fatty acid and isolated from the black sponge, genus Halichondria, on cytosolic Ca2+ concentration ([Ca2+]i) and tension developed in porcine coronary arterial strips loaded with fura-2. 2. Both in the presence (1.25 mM) and absence of extracellular Ca2+, OA (over 10(-6) M) induced a concentration-dependent, slow and progressive increase in tension. Calcium removal had no effect on the maximum level of tension, time between application of the drug and the onset of tension, or the time required to reach the maximum tension. However, there was a slight concentration-dependent increase in [Ca2+]i, only in the presence of extracellular Ca2+. 3. At a lower concentration that did not cause contraction or increase [Ca2+]i, OA (10(-6) M) inhibited tension development but not the Ca2+ transient on readmission of Ca2+ in 118 mM K+-depolarizing solution. OA inhibited the maximum levels of the developed tension, without affecting the KD value (598 +/- 204 nM for control vs 678 +/- 464 nM after OA treatment) or the Hill coefficient (1.78 +/- 0.10 for control vs 1.98 +/- 0.47 for OA treatment). 4. It is concluded that high concentrations of OA induce a contraction independent of extracellular Ca2+ and without any changes in [Ca2+]i. Lower concentrations of OA inhibit the Ca2+-dependent contractions. The lack of effect on KD values suggests that the [Ca2+]i-sensitivity of the contractile apparatus is not affected by this inhibition of contraction.

Animals↗
PubMed1989

Low-voltage-activated calcium current in rat aorta smooth muscle cells in primary culture.

1. Electrical and pharmacological properties of the low-voltage-activated Ca2+ current (ICa, LVA) in rat aorta smooth muscle cells (SMC) in primary culture were examined, particularly in comparison with the high-voltage-activated Ca2+ current (ICa, HVA). Both types of Ca2+ currents were recorded in external solution containing 20 mM-Ca2+, using the whole-cell voltage-clamp technique. 2. ICa, LVA was evoked by step depolarizations to potentials more positive than -60 mV from a holding potential of -100 mV, and reached a peak in the current-voltage (I-V) relationship around -30 mV. ICa, HVA was activated at -20 mV, and reached a peak at +20 mV. 3. The intracellular dialysis of 5 mM-F- irreversibly suppressed ICa, HVA, with time, while it has little effect on the ICa, LVA. The ICa, LVA could be separated from the ICa, HVA by either selecting the holding and test potential levels or by perfusing intracellularly with F-. 4. The ratio of peak amplitude of Ba2+, Sr2+ and Ca2+ currents in the respective I-V relationship was 1.6:1.2:1.0 for high-voltage-activated Ca2+ channels and was 1.0:1.4:1.0 for low-voltage-activated ones. 5. The inactivation phase of ICa, HVA was fitted by a sum of double-exponential functions, the time constants of which were larger when the current was carried by Ba2+ than by Ca2+. The inactivation time course of ICa, LVA was fitted by a single-exponential function, and the time constant was practically the same when the current was carried by Ba2+ or by Ca2+. Activation and inactivation processes of ICa, LVA were potential-dependent. 6. The steady-state inactivation curve of ICa, LVA was fitted by the Boltzmann equation, having a mid-potential of -80 mV and a slope factor of 5.0. The recovery time course from steady-state inactivation was fitted by a sum of two exponential functions. The time constants of the faster phase were 230 and 380 ms, and those of slower phase were 2.8 and 1.8 s at the repolarization potentials of -120 and -100 mV, respectively. 7. The amplitude of ICa, LVA depended on the external Ca2+ concentration ([Ca2+]o), approaching saturation at 95 mM [Ca2+]o. 8. Various polyvalent cations blocked both types of Ca2+ current reversibly in the order (IC50 in M): La3+ (8 x 10(-8)) greater than Cd2+ (6 x 10(-6)) greater than Ni2+ (1 x 10(-5)) greater than Zn2+ (2 x 10(-5)) for ICa, HVA, and La3+ (6 x 10(-7)) greater than Zn2+ (3 x 10(-5)) greater than Cd2+ (4 x 10(-4)) greater than Ni2+ (6 x 10(-4)) for ICa, LVA.(ABSTRACT TRUNCATED AT 400 WORDS)

4-Aminopyridine↗
PubMed1989

Histamine-induced calcium transients in vascular smooth muscle cells: effects of verapamil and diltiazem.

We investigated the effects of verapamil and diltiazem on histamine-induced Ca2+ transients in vascular smooth muscle. 1) With the use of quin2 microfluorometry, cytosolic Ca2+ concentrations were directly measured in cultured vascular smooth muscle cells of the rat aorta. In the presence of extracellular Ca2+, histamine induced an elevation of cytosolic Ca2+ concentrations of a peak and plateau type. The peak component is due to a release of Ca2+ from cellular store sites, and the plateau component depends on extracellular Ca2+. Verapamil and diltiazem inhibited the plateau component, and the 50% inhibitive concentration (IC50) of verapamil and diltiazem for 10 microM histamine was 0.09 and 0.18 microM, respectively. Only at high concentrations did verapamil (IC50 = 8.7 microM) and diltiazem (IC50 = 95.7 microM) inhibit the Ca2+ release from the cellular store sites, as induced by 10 microM histamine. 2) Histamine, verapamil, and diltiazem competed with [3H]mepyramine for binding to the porcine aortic membranes, the order of potency being verapamil (Ki = 7.1 microM) greater than histamine (Ki = 18 microM) greater than diltiazem (Ki = 114 microM). From these results, we conclude that verapamil and diltiazem strongly inhibit the histamine-mediated, extracellular Ca2+-dependent intracellular [Ca2+] increase. In addition, verapamil and diltiazem seem to inhibit the release of Ca2+ from intracellular store sites, only at high concentrations, and probably by competing with histamine for binding to the H1-receptor. The inhibitory effects of Ca2+ antagonists on the histamine-induced contraction or spasm of vascular smooth muscle may well relate to these mechanisms.

Aminoquinolines↗
PubMed1989

Effects of nicorandil on cytosolic calcium concentrations in quin2-loaded rat aortic vascular smooth muscle cells in primary culture.

We made use of quin2 microfluorometry to observe the effects of nicorandil (2-nicotinamidoethyl nitrate) on cytosolic Ca++ concentrations [( Ca++]i) in rat aortic vascular smooth muscle cells in primary culture. Regardless of whether cells were at rest, in a state of Ca++-depletion or at K+-depolarization, nicorandil rapidly and dose-dependently decreased [Ca++]i to the lower steady-state level. Nicorandil dose-dependently inhibited norepinephrine-induced Ca++ transients in physiological salt solution containing 1 mM Ca++. Nicorandil accelerated the reduction of [Ca++]i observed when the cells were exposed to Ca++-free solution. When the cells were treated with nicorandil in Ca++-free solution, Ca++ transients induced by the first application of caffeine were little affected, but those induced by subsequent repetitive caffeine applications were reduced strongly and progressively. In contrast, pretreatment with nicorandil markedly inhibited Ca++ transients induced by the first application of norepinephrine, in Ca++-free solution. These effects of nicorandil on [Ca++]i and Ca++ transients were similar to those seen with nitroglycerin. The denitrated compound of nicorandil, N-(2-hydroxyethyl)nicotinamide, had no such effect. Thus, it is apparently the nitrate moiety of the chemical structure by which nicorandil actively and strongly reduces [Ca++]i in vascular smooth muscle cells. The reduction of [Ca++]i by nicorandil may result in a decrease in Ca++ in the norepinephrine-sensitive store; hence, the reduction of [Ca++]i elevation by norepinephrine.

Aminoquinolines↗
PubMed1989

Selective blockade of alpha 1-adrenoceptors of porcine vascular smooth muscle cells by bunazosin.

The characteristics of bunazosin binding to alpha-adrenoceptors in the porcine aortic membranes were investigated using [3H]prazosin and [3H]yohimbine binding assays to identify alpha 1- and alpha 2-adrenoceptors, respectively. The extent of the inhibition (Ki values) of [3H]prazosin binding to alpha 1-adrenoceptors induced by bunazosin was 0.29 nmol/l and about the same as that induced by prazosin (Ki = 0.10 nmol/l). The Ki value of bunazosin inhibition of [3H]yohimbine binding to alpha 2-adrenoceptors was 350 nmol/l. There was over a 1000-fold difference in Ki value for bunazosin between alpha 1-and alpha 2-adrenoceptors. Thus, bunazosin is a highly alpha 1 selective agent in vascular smooth muscle.

Adrenergic alpha-Antagonists↗
PubMed1988

Does endothelin mobilize calcium from intracellular store sites in rat aortic vascular smooth muscle cells in primary culture?

In the presence of endothelin, there was a rapid increase in the 45Ca++ efflux from primary cultured rat vascular smooth muscle cells, both in physiological salt solution and in calcium free medium containing 2 mM EGTA. The 45Ca++ influx was not affected. The endothelin-induced, transient increase in cytosolic calcium concentration is probably mainly due to release of calcium from the intracellular store in vascular smooth muscle cells.

Animals↗
PubMed1988

Ca-activated K channel in cultured smooth muscle cells of rat aortic media.

Electrical and pharmacological properties of the single Ca-activated K channel in cultured smooth muscle cells (SMC) of the rat aorta were studied with the patch-clamp technique. The Ca-activated K channel had a slope conductance (gamma K) of 135 +/- 2 pS (mean +/- SE; n = 5) in symmetrical 142 mM K solutions. The reversal potentials show a 56-mV change for a 10-fold change in the external K concentration. Probability of the channel opening increased when the intracellular Ca concentration ([Ca]i) was increased over 10(-7) M or the membrane was depolarized. The channel was blocked by either external tetraethylammonium (TEA, 10-30 mM) or by internal Ba (1-5 mM). Channel activities were characterized by burst-like openings. Open-time histogram was fitted with a single exponential (tau = 1.3 ms at +10 mV and 10(-7) M [Ca]i), whereas the closed-time histogram was fitted with two exponentials (tau 1 = 0.7 ms and tau 2 = 111 ms). The permeability ratio for monovalent cations calculated with the Goldman-Hodgkin-Katz equation was K:Rb:Na = 1:0.7: less than 0.01. We conclude from these observations that the Ca-activated K channel in cultured SMC of the rat aorta is characterized by a middle size gamma K, activation by [Ca]i increase and depolarization, relatively low sensitivity to TEA, and high selectivity for K ions.

Animals↗
PubMed1988

Cyclic AMP modulates Ca-activated K channel in cultured smooth muscle cells of rat aortas.

Effects of adenosine 3',5'-cyclic monophosphate (cAMP) on single Ca-activated K current (IK(Ca)) in cultured smooth muscle cells of the rat aorta were investigated with the patch-clamp technique. In cell-attached patch configurations, extracellular application of isoproterenol (10(-5) M) increased the Ca-activated K currents. The increase in the currents was due to an increase in the probability of channel openings (Po). Neither unit conductance nor the maximum number of the channel in the patch was affected by the drug. The effects were inhibited by adding propranolol (10(-6) M). The extracellular application of forskolin (10(-5) M) or dibutyryl cAMP (10(-4) M) mimicked the effects of isoproterenol. In inside-out patch configurations, activated cAMP-dependent protein kinase (A kinase) in the bathing solution increased the sensitivity of the Ca-activated K channels to intracellular free calcium concentration ([Ca]i) and enhanced Po. Kinetic analyses of the IK(Ca) showed that cAMP-dependent phosphorylation of the Ca-activated K channels significantly reduced the mean closed time between bursting openings. We conclude from these observations that the Ca-activated K channels in aortic cells may increase Po through cAMP-dependent phosphorylation.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1988

Quin2 microfluorometry and effects of verapamil and diltiazem on calcium release from rat aorta smooth muscle cells in primary culture.

We investigated the effects of the Ca2+ antagonists diltiazem and verapamil on release of Ca2+ from intracellular store sites of rat aorta vascular smooth muscle cells in primary culture. Using the microfluorometry of Ca2+-indicator dye quin2, relative changes in cytosolic Ca2+ concentration could be measured. In the presence of 1 mM extracellular Ca2+, both diltiazem (IC50, 0.31 microM) and verapamil (IC50, 0.47 microM) dose-dependently inhibited elevations in the cytosolic Ca2+, as induced by depolarization of the plasma membrane with high extracellular K+. In the absence of extracellular Ca2+, caffeine and high extracellular K+ induced transient and dose-dependent elevations of the cytosolic Ca2+, and these elevations were not inhibited by either diltiazem or verapamil. Norepinephrine also induced a transient and dose-dependent elevation of cytosolic Ca2+ in the absence of extracellular Ca2+. However, this elevation was inhibited by verapamil and diltiazem (when the norepinephrine concentration was 10(-5) M, IC50 for verapamil and diltiazem was 4.0 and 24.9 microM, respectively). Thus, while verapamil and diltiazem may have no direct effect on the release of Ca2+ from the depolarization- and the caffeine-sensitive intracellular Ca2+ storage sites, the agents do seem to inhibit the adrenoceptor-mediated Ca2+ release mechanism in vascular smooth muscle cells.

Aminoquinolines↗
PubMed1987

8-Bromoguanosine 3':5'-cyclic monophosphate decreases intracellular free calcium concentrations in cultured vascular smooth muscle cells from rat aorta.

The effects of 8-bromoguanosine 3':5'-cyclic monophosphate (8-Br cGMP) on intracellular free calcium concentrations ([Ca2+]i) in cultured rat aortic vascular smooth muscle cells (VSMCs) loaded with fura-2 were recorded microfluorometrically. Irrespective of whether VSMCs were at rest (in 5 mM K+ PSS), under Ca2+ depletion (in Ca2+-free medium for 10 min) and K+ depolarization (in high K+ PSS), [Ca2+]i was actively reduced and reached a new and lower steady-state level with the application of 8-Br cGMP. This may be the first and direct evidence that cGMP, a putative mediator of various vasodilators, actively reduces [Ca2+]i in VSMCs.

Animals↗
PubMed1987

The norepinephrine-sensitive Ca2+-storage site differs from the caffeine-sensitive site in vascular smooth muscle of the rat aorta.

Using microfluorometry of quin 2, a Ca2+-sensitive dye, we characterized the release and uptake of Ca2+ by the norepinephrine-sensitive Ca2+-storage site and the caffeine-sensitive one. The norepinephrine-sensitive Ca2+-storage site was readily depleted in Ca2+-free medium and almost completely replenished by loading with 1.0 mM Ca2+ solution for 3 min, whereas the caffeine-sensitive site was scarcely affected. Furthermore, norepinephrine has little effect on the caffeine-sensitive Ca2+-storage site in Ca2+-free medium, and vice versa. We conclude that the location and mechanisms of release and uptake of Ca2+ of these two Ca2+-storage sites differ in the case of rat aortic vascular smooth muscle cells in primary culture.

Aminoquinolines↗
PubMed1987

Serotonin-induced cytosolic free calcium transients in cultured vascular smooth muscle cells.

Serotonin induced a transient elevation in the levels of cytosolic calcium in cultured rat vascular smooth muscle cells. Ketanserin, a selective antagonist of serotonin 2 receptors, dose-dependently inhibited the elevation of cytosolic calcium induced by serotonin, and ultimately unmasked a serotonin-induced decrease in the levels of cytosolic calcium. These observations show that serotonin has direct and dual effects, that is, it increases and decreases cytosolic free calcium concentrations in vascular smooth muscle cells, in culture. Knowledge of such events is important because serotonergic inhibitors may prove to be useful drugs for treating clinical hypertension and vasospastic disorders.

Aminoquinolines↗
PubMed1987

Adenosine decreases intracellular free calcium concentrations in cultured vascular smooth muscle cells from rat aorta.

Using an intracellularly trapped dye, quin 2, effects of adenosine on intracellular free calcium concentrations ([Ca2+]i) were recorded, microfluorometrically, using rat aortic medial vascular smooth muscle cells (VSMCs) in primary culture. Regardless of whether cells were at rest (in 5 mM K+), at K+-depolarization (in 55 mM K+) or at Ca2+ depletion (in Ca2+-free media), adenosine induced a rapid reduction of [Ca2+]i, following which there was a gradual increase to pre-exposure levels, in cells at rest and in the case of Ca2+ depletion. Only when the cells were depolarized (55 mM K+) did adenosine induce a new steady [Ca2+]i level, lower than the pre-exposure value. These findings indicate that decrease in [Ca2+]i by adenosine is one possible mechanism involved in the adenosine-mediated vasodilatation, and that adenosine decreases [Ca2+]i by direct extrusion, by sequestration, or by inhibiting the influx of Ca2+ into VSMCs.

Adenosine↗
PubMed1987

Ketanserin blocks alpha 1-adrenoceptors of porcine vascular smooth muscle cells.

The effect of ketanserin on alpha-adrenoceptors was studied in membrane preparations of the porcine aorta using [3H]prazosin and [3H]yohimbine binding assays to identify alpha 1- and alpha 2-adrenoceptors. Ketanserin bound to alpha-adrenoceptors and the Ki value of ketanserin for alpha 1-adrenoceptors was 8.3 nM, a value practically equal to that of phentolamine (Ki = 7.2 nM). The Ki value of ketanserin for alpha 2-adrenoceptors was 3.3 microM. Thus, at the doses prescribed clinically, ketanserin blocks alpha 1- but not alpha 2-adrenoceptors of porcine vascular smooth muscle.

Adrenergic alpha-Antagonists↗
PubMed1987

Transmural anoxic wave front and regional dysfunction during early ischemia.

The relative time courses of early changes in myocardial metabolism and function during anoxia, global ischemia, and regional ischemia were compared in isolated rat hearts. Transmural anoxic wave front was determined with NADH fluorescence photography, and oxygen saturation of myoglobin and dynamic systolic wall thickening were measured with spectrophotometry of light transmitted through the left ventricular free wall. In all three treatments, anoxic wave front first appeared in the subendocardium and reached the epicardial half of the myocardium in 10 s, when oxygen saturation of myoglobin decreased by 50% and tissue ATP and creatine phosphate remained at aerobic levels. During this period, systolic wall thickening decreased gradually in anoxia and global ischemia, whereas a marked decrease in systolic wall thickening and appearance of dyskinesia (wall thinning) occurred in regional ischemia. Thus the early extension of anoxic wave front and metabolic changes are similar with all three treatments, and dyskinesia, observed only in case of regional ischemia, occurs when the inner half is ischemic or anoxic.

Animals↗
PubMed1987

Characteristics of adrenoceptors and [3H]nitrendipine receptors of porcine vascular smooth muscle: differences between coronary artery and aorta.

Characteristics of the bindings of [3H](-)dihydroalprenolol, [125I](-)iodocyanopindolol, [3H]prazosin, [3H]yohimbine, and [3H]nitrendipine to porcine coronary membranes were investigated and the results compared with studies of porcine aortic membranes. In the equilibrium binding study carried out in sarcolemma-enriched fractions, there were no major differences in the Kd values of these radioligands between coronary artery and aorta. However, the densities of beta-, alpha 1-, and alpha 2-adrenoceptors and [3H]nitrendipine receptors of coronary artery were 258, 12, 12, and 561 fmol/mg protein, respectively, while those of aorta were 37, 525, 1,000, and 215 fmol/mg protein. beta-Adrenergic agonists competed with [3H](-)dihydroalprenolol binding sites in coronary artery, the order of potency being (-)isoproterenol greater than (-)norepinephrine greater than (-)epinephrine greater than (+)isoproterenol. In case of aorta, the order was (-)isoproterenol greater than (-)epinephrine greater than (-)norepinephrine. The competition by (+/-)bisoprolol (beta 1-selective antagonist) and ICI 118,551 (beta 2-selective antagonist) for [125I](-)iodocyanopindolol binding sites in coronary artery resulted in nonlinear Hofstee plots (beta 1:beta 2 = 90%:10%). In case of aorta, linear Hofstee plots were obtained. From these results, we conclude that coronary beta-receptors in pigs are predominantly of beta 1-type, while those of aorta are of beta 2-type; regarding the relative population of adrenoceptors, coronary artery is beta-dominant (beta/alpha = 11), while aorta is alpha-dominant (beta/alpha = 0.02); compared with alpha-adrenoceptors, coronary artery has a greater number of [3H]nitrendipine binding sites (nitrendipine/alpha-adrenoceptor = 23) than aorta (nitrendipine/alpha-adrenoceptor = 0.14).

Animals↗