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Biomedical subjects

J Morikawa

Publications and source records attributed to J Morikawa.

At least 37 records · Page 2Linked to original sources
PubMed1997

Dopamine stimulation of cardiac beta-adrenoceptors: the involvement of sympathetic amine transporters and the effect of SKF38393.

1. Mechanisms underlying beta-adrenoceptor stimulation by dopamine were examined on guinea-pig Langendorff-perfused hearts and isolated cells from the right atrium, by using the chronotropic effects and the enhancement of L-type Ca2+ current (ICa,L) in the presence of prazosin as indicators of beta-adrenoceptor stimulation. Dopamine-induced overflow of noradrenaline (NA) concentrations was measured by high-performance liquid chromatography. 2. Dopamine caused positive chronotropic effects with an EC50 of 2.5 microM and induced NA overflow with a similar EC50 (1.3 microM). The chronotropic effect of dopamine was abolished by bisoprolol (1 microM). 3. The effects of dopamine were maintained during prolonged application, whereas the effects of tyramine faded with time. Dopamine (3 microM) restored the chronotropic effects and the NA release suppressed by pretreatment with tyramine, suggesting a de novo synthesis of NA during the exposure to dopamine. 4. Dopamine (3 microM)-induced NA release was not affected by tetrodotoxin, omega-conotoxin, rauwolscine, ICI118551 or sulpiride, but was inhibited by desipramine, a NA uptake inhibitor (IC50 approximately 1 microM). It was also not affected by GBR12909 and bupropion, dopamine uptake inhibitors in the central nervous system. 5. SKF38393, a D1 receptor partial agonist, potently inhibited the 3 microM dopamine-induced release of NA (IC50 approximately 0.1 microM). D1 receptors are not involved in the DA-induced release of NA, since SCH23390 (3 microM), a potent D1 antagonist, inhibited the NA release only slightly, and dihydrexidine (1 microM) and chloro-APB (1 microM), full D1 agonists, caused no significant NA release. 6. SKF38393 inhibited tyramine-induced overflow of NA, and potentiated the field stimulation-induced NA release. SKF38393 and desipramine retarded the decay of the stimulation-induced tachycardia in a similar manner. These results indicate that SKF38393 is a potent monoamine transport inhibitor and a useful tool for the functional evaluation of indirectly-acting sympathomimetic agonists in the heart. In the presence of SKF38393 (10 microM), dopamine at 1 microM showed no chronotropic effect. 7. Voltage clamp experiments with isolated atrial cells revealed that dopamine is a weak partial agonist. The EC50 for ICa,L stimulation by dopamine was high (13 microM). As a result, dopamine at 1 microM did not affect ICa,L. Bisoprolol abolished the stimulation of ICa,L by dopamine (30 microM), and dihydrexidine (1 microM) did not affect ICa,L. 8. It was concluded that the cardiac effects of dopamine at clinically relevant concentrations (< 1 microM) result almost exclusively from the indirect effect of beta adrenoceptor stimulation, involving the release of NA from sympathetic nerve terminals. The roles of the direct stimulation of beta adrenoceptors by dopamine at these concentrations and the stimulation of postjunctional D1 receptors seem negligible. The desipramine- and SKF38393-sensitive monoamine transporter mediates the release of NA.

2,3,4,5-Tetrahydro-7,8-dihydroxy-1-phenyl-1H-3-ben↗
PubMed1997

Biochemical and immunochemical properties of modified human neuron-specific enolase.

125I-Labeled recombinant human neuron-specific enolase (R-NSE) was inadequate for RIA as a labeled antigen. The binding activity of labeled R-NSE to the antibody was markedly decreased. To supplement this defect and facilitate purification, we constructed two R-NSE derivatives, Y-NSE (one tyrosine residue was added at the N-terminal of R-NSE) and Y-NSE.H6 (six histidine residues were further added at the C-terminal of Y-NSE). The biochemical and immunochemical characteristics of these R-NSE derivatives were essentially the same to those of R-NSE. These derivatives were useful not only as standards for enzyme immunoassay (EIA), but also as labeled antigens for RIA. These results clearly indicate that the reactivity of these modified NSEs to anti-NSE antibody is almost equivalent to that of human brain gammagamma-enolase (B-NSE), and that even if the modified NSEs are labeled, they retain their binding affinities to antibodies in contrast to R-NSE.

Base Sequence↗
PubMed1997

Muscarinic inhibition of basal L-type calcium current in pacemaker cells from the rabbit atrioventricular node.

Effects of acetylcholine (ACh) on the L-type calcium current were examined in isolated atrioventricular nodal cells that exhibited spontaneous contractions. ACh (0.1 to 10 microM) inhibited basal calcium current dose-dependently. This inhibition was eliminated by dialysis with 8Br cAMP or cAMP-dependent kinase inhibitory peptide. Both extracellular N-ethylmaleimide 50 microM and intracellular GDPssS 0.2 mM abolished the ACh effect. Dialysis with cGMP or NG-monomethyl-L-arginine did not significantly affect ACh inhibition of basal calcium current. Similarly, cGMP-dependent protein kinase inhibitor KT5823 (1 microM) and the type II phosphodiesterase inhibitor erythro-9-(2-hydroxy-3-nonyl) adenine (30 microM) did not attenuate the ACh effect. Therefore, ACh inhibits the basal calcium current in the atrioventricular node mainly by suppressing cAMP synthesis through the inhibitory GTP-binding protein.

Acetylcholine↗
PubMed1996

Modulation of L-type Ca current by denopamine, a nonparenteral partial beta 1 stimulant, in rabbit ventricular cells.

The effects of denopamine, a nonparenteral partial beta agonist which is used clinically in Japan, on the L-type Ca2+ current (ICa) were examined in rabbit ventricular cells. Denopamine stimulated basal ICa with a maximum response of +33.2% and a concentration for half-maximal response (EC50) of 0.039 microM. The maximum response of ICa was only a quarter of that induced by isoprenaline (ISO), while 10 microM denopamine elicited 70-75% of the maximum inotropic response in the papillary muscle preparations. The denopamine stimulation of ICa was abolished by selective beta 1 antagonists (atenolol or bisoprolol). Pretreatment with forskolin or dialysis with cAMP also abolished the stimulation. Denopamine, in turn, inhibited ISO-stimulated ICa. This inhibition was not affected by pretreatment with pertussis toxin or prazosin. The presence of denopamine at various concentrations caused a rightward shift in the concentration/response curve for ISO stimulation of ICa. The Schild plot for this effect had a slope of 0.99 and Kp of 0.20 microM. In the presence of guanosine-5'-O-(3-thiotriphosphate) (GTP gamma S) (0.5 mM) in the pipette, denopamine (10 microM) stimulated the ICa to 86 +/- 5% of the maximum response induced by ISO. These findings indicate that denopamine modulates ICa exclusively through the beta 1 adrenoceptor-adenylate cyclase pathway, that the stimulatory GTP-binding protein regulates the agonistic potency of denopamine, and that the signal from the beta 1 adrenoceptors is amplified between ICa and the tension development, which would contribute to the spare capacity of beta adrenoceptors.

Adenylyl Cyclases↗
PubMed1996

Purification of pepsinogens from human urine and electrophoretic analysis by caseogram print.

Pepsinogen (PG) A and C were purified from human urine, and analyzed by a highly sensitive detection method, "caseogram print". Purification was achieved by a series of conventional chromatographies and FPLC. A relatively large amount (13.2 mg) of PGA was purified from about 20 liters of urine. Purified PGA was separated by a Mono-Q column into each of its isozymogens. The elution order (PGA-5, 4+3, 2) corresponded to the order of electrophoretic migration. Although the concentration of urinary PGC was very low, a trace amount was purified and visualized by electrophoresis. The urinary and mucosal PGCs migrated at the same position, and urinary PGC was detected as two isozymogens similarly to mucosal PGC, suggesting that urinary and mucosal PGCs may be essentially identical.

Chromatography, Gel↗
PubMed1996

Modification of human neuron-specific enolase for application to radioimmunoassay.

A recombinant human neuron-specific enolase (R-NSE), isolated from Escherichia coli, could not be used in an RIA system because of instability upon labeling. To apply R-NSE to RIA and to simplify the purification procedure, the N- and C-terminals of R-NSE were modified by tyrosine- and histidine-tagging, respectively. SY-NSE, containing one additional tyrosine residue, was obtained from both soluble and insoluble fractions. More derivatives tagged by two or four tyrosine residues were expressed, but only in the insoluble fraction. SY-NSE and SY-NSE.H6 (containing six histidine residues at C-terminal of SY-NSE) purified from the soluble fraction were applicable to the RIA system, indicating that the addition of a tyrosine residue at the terminal is effective if the antigen is unstable during labeling.

Base Sequence↗
PubMed1996

Delineation of premature P waves on four-dimensional electrocardiography, a new display of electrical forces by computer techniques.

This study investigated the feasibility of four-dimensional electrocardiography (4-D ECG), a new display in which the vector loop was rotated and scanned along a timed axis to overcome the shortcomings of vectorcardiography (VCG). The subjects consisted of 38 patients with premature atrial complexes and 30 controls. The orthogonal Frank electrocardiograms were rotated three-dimensionally according to the right-hand rectangular coordinate system and scanned along a timed axis. The P wave delineation score, signifying good agreement with the intraobserver and interobserver variability, was significantly higher in 4-D ECG than those in the orthogonal leads or those on the transverse and frontal projections (P < 0.001). The authors measured the premature P loop areas as viewed from 361 directions. P loop areas were best delineated when viewed from cranial directions of 42.6 +/- 34.0 degrees and from rightward directions of 11.3 +/- 30.7 degrees. Adequate cranial rotation followed by scanning along a timed axis will maximally delineate premature atrial signals and provide comprehensive visualization of electrical forces.

Atrial Premature Complexes↗
PubMed1995

Ethanol inhibition of Ca2+ and Na+ currents in the guinea-pig heart.

The effects of ethanol on L-type Ca2+ and fast Na+ currents (ICa and INa, respectively) were examined using the whole-cell patch-clamp experiments on guinea-pig ventricular cells. At a clinically relevant concentration of 24 mM, ethanol slightly but significantly shortened the action potential duration, and reduced the ICa by 7 +/- 4% (mean +/- S.D.). This concentration of ethanol did not affect INa, but a lethal concentration of ethanol (80 mM) significantly inhibited INa by 13 +/- 5%. The voltage dependence of INa activation was not affected by ethanol, whereas the inhibitions of ICa by 80 mM ethanol and INa by 240 mM were both accompanied by a several mV shift in the channel availability curve toward more negative potentials, suggesting that the channels in the inactivated state are more susceptible to ethanol. The ICa inhibition by ethanol at clinically relevant concentrations could contribute to a negative inotropic effect, action potential shortening and development of arrhythmias, while the pathophysiological significance of ethanol inhibition of INa seems less important.

Action Potentials↗
PubMed1995

Electrophysiological effects of changrolin, an anti-arrhythmic agent derived from Dichroa febrifuga, on guinea-pig and rabbit heart cells.

1. The electrophysiological effects of changrolin (CRL), a Chinese anti-arrhythmic drug derived from a traditional antimalarial plant, were examined using the whole-cell patch-clamp method on single cells isolated from guinea-pig and rabbit hearts. 2. At a clinically relevant concentration of 50 mumol/L changrolin inhibited ICa by 19.3 +/- 6.0% and 17.3 +/- 2.6% in guinea-pig and rabbit ventricular cells, respectively. The voltage-dependent channel availability curve was not affected. The CRL effect was enhanced to a small extent during a repetitive stimulation at 2 Hz. 3. INa was resistant to CRL and the channel availability curve was also unaffected. A small use-dependent inhibition was observed only when the INa was elicited at 5 Hz in the presence of 300 mumol/L CRL. 4. At 50 mumol/L, CRL did not affect the time-independent inward rectifier and the delayed rectifier K+ currents (IK1 and IK, respectively), but inhibited the transient outward current (ITO) by 17.7 +/- 2.4%. Changrolin significantly shortened the action potential duration in both guinea-pig and rabbit ventricular cells. 5. In conclusion, CRL inhibits ICa and ITO but has little effect on INa.

Action Potentials↗
PubMed1995

Decreased sensitivity to beta-adrenergic stimulation of the ventricular cells isolated from the spontaneously hypertensive rat heart.

1. The stimulatory effects of isoproterenol on the L-type Ca2+ current (ICa) were compared between the control (WKY) and hypertensive (SHR) rat heart cells, using the patch-clamp method. 2. The current density and the shape of the current-voltage relationship for ICa were not different between the two groups. However, the maximal percentage increase in response to isoproterenol was smaller in SHR (+91% in SHR and +81% in WKY), and the ED50 was significantly higher in SHR (0.081 mu mol/L in SHR and 0.020 mu mol/L in WKY). IBMX, a potent phosphodiesterase inhibitor, significantly increased the isoproterenol-stimulated ICa in SHR, but not in WKY. These results suggest an impaired cAMP production in SHR heart cells.

1-Methyl-3-isobutylxanthine↗
PubMed1995

Angiotensin II inhibition of L-type Ca2+ current in sinoatrial node cells of rabbits.

The actions of angiotensin II (ANG II) were examined in the spontaneously active cells isolated from the rabbit sinoatrial node, using the nystatin-permeabilized, whole cell, patch-clamp method. At 30 nM, ANG II significantly lowered the spontaneous firing rate of the action potentials from 212 +/- 21 to 172 +/- 32 beats/min, with a concomitant reduction in the action potential amplitude. The voltage-clamp experiments showed that ANG II inhibited the L-type Ca2+ current (ICa) with a dissociation constant (Kd) of approximately 4 nM and a maximal inhibition of 30%. The inhibition was blocked by an AT1-receptor antagonist CV11974. Acetylcholine (ACh) at 10 microM reduced the ICa by 42 +/- 12%, and ANG II did not cause any further inhibition in the presence of ACh. At 100 nM, ANG II reduced the ICa by only 12% in the presence of 2 microM isoproterenol, and a similar inhibition was observed with 0.1 microM ACh. ANG II did not affect the dibutyryl adenosine 3',5'-cyclic monophosphate-stimulated ICa. Protein kinase C activator 12-O-tetra-decanoylphorbol-13-acetate did not mimic ANG II in the effects on ICa, and preincubation of the cells with calphostin C, a protein kinase C inhibitor, did not attenuate the ANG II effect. ANG II exerts a negative chronotropic effect in the pacemaker cells as its direct action through a pathway involving adenosine 3',5'-cyclic monophosphate-dependent protein kinase.

Acetylcholine↗
PubMed1995

Conversion of cathepsin E to enzymatic unstable form in gastric cancer cells.

A new acid proteinase in human gastric cancer, named medium moving proteinase (Med.P), was found also in a gastric cancer transplanted into nude mouse. However, Med.P disappeared when the samples prepared from gastric cancer tissues were left for over 4 weeks at -80 degrees C, whereas the activity of cathepsin E (CE) increased. When these samples were reduced by dithiothreitol (DTT), Med.P appeared again and the CE activity decreased. These phenomena, revealed by electrophoretic analyses, indicated that Med.P is a monomeric form of CE (mono-CE). At weakly alkaline pH and after heating, mono-CE appeared to be more unstable than CE. These results indicated that CE assume an enzymatically unstable monomeric form in cancer cells.

Animals↗
PubMed1995

Does dopamine act on myocardial cells?

We examined the electrophysiological effects of dopamine on the single myocardial cells isolated from the rat and rabbit heart. Dopamine at a concentration of 1 or 10 microM did not affect the L-type Ca2+ current (ICa) or the transient outward current (ITO) in rat ventricular, rabbit atrial, ventricular, and sinoatrial node cells. It did not induce any detectable change in the action potential configuration of the rabbit ventricular cells either. We conclude that dopamine does not directly act on myocardial cells at least in terms of the electrophysiological properties.

Action Potentials↗
PubMed1994

Modulation of sodium current by lactate in guinea pig ventricular myocytes.

OBJECTIVE: The aim was to elucidate whether or not lactate modifies the fast sodium current (INa) in cardiac cells. METHODS: A tight seal whole cell clamp technique was used to record the action potentials and INa in single ventricular cells from the guinea pig heart. RESULTS: In voltage clamp experiments, superfusion with 20 mM lactate shifted both the normalised conductance (gNa)-voltage relationship and the channel availability curve toward hyperpolarisation by approximately 4 mV, but did not affect the maximum conductance (gNa,max). In the test solution containing only CaCl2 as the main divalent component, 20 mM lactate reduced the ionised calcium concentration from 1.02 to 0.84 mM. When the calcium concentration was kept constant by the addition of CaCl2 into the lactate containing solution the lactate effect was nullified. However, a change in the calcium concentration from 1.0 to 0.84 mM without lactate induced a 4 mV negative shift of the channel availability curve. Current clamp experiments in Tyrode solution showed that 20 mM lactate shifted the threshold for the action potential upstroke by 2.5-3 mV, in accordance with the voltage clamp experiments. CONCLUSIONS: Lactate modifies INa of ventricular myocytes by shifting its kinetics toward hyperpolarisation. This shift seems to be caused exclusively by a decrease in the ionised divalent cation concentrations and a resultant change in the negative surface charge of the sarcolemma.

Action Potentials↗
PubMed1994

A case of lipoprotein glomerulopathy successfully treated with probucol.

A 54-year-old woman presented proteinuria and severe hyperlipoproteinemia, She had type III hyperlipoproteinemia with the phenotype E2/2. Renal biopsy specimen revealed lipoprotein thrombi within the glomerular capillary lumina without tubulointerstitial lesion and deposition of IgA in the mesangium and peripheral capillary loops, indicating lipoprotein glomerulopathy with IgA nephropathy. Treatment with probucol resulted in the improvement of both hyperlipoproteinemia and proteinuria with the disappearance of the lipoprotein thrombi in the glomerular capillary without any change in the mesangial IgA deposition. The data suggest that probucol may be effective against the early stage of lipoprotein glomerulopathy with less lipoprotein thrombi and without tubulointerstitial damage.

Female↗
PubMed1994

Electrophoretic analysis of a gastric cancer-associated acid proteinase using a highly sensitive detection system.

A highly sensitive detection system for acid proteinase separated on polyacrylamide gel was established. This system consisted of two-dimensional electrophoresis, combined with isoelectric focusing and polyacrylamide gel electrophoresis, and casein clotting (caseogram). Human urine, serum and gastric tissues obtained from normal individuals and gastric cancer patients were analyzed using this system. The previous electrophoretic method was not sufficiently sensitive to detected small amounts of pepsinogen (PG) C in normal urine. However, the new rapid and sensitive method clearly revealed its presence. In gastric tissue containing cancer cells, an additional proteinase, which was not present in normal tissue, was detected and named medium moving proteinase (MMP). MMP resembled PGs in alkaline stability rather than the non-PG proteinase, slow moving proteinase (SMP).

Adult↗