Slow recovery of conduction velocity from use dependent inhibition induced by quinidine in guinea pig ventricular myocardium.

STUDY OBJECTIVE: The aim was to investigate whether the use dependent effects of antiarrhythmic drugs on the Na+ current could be applied to explain their effects on impulse conduction. DESIGN: Trains of rapid stimuli were applied to guinea pig papillary muscles via an electrode in the presence of quinidine (20 and 60 mumol.litre-1), and the conduction velocity was determined from the time difference between two signals of the maximal rate of rise (dV/dtmax) of the action potentials at two separate sites. The relationship of the time constants of the onset and recovery from the use dependent inhibition induced by quinidine was determined for the dV/dtmax and the conduction velocity. EXPERIMENTAL MATERIAL: Six male Hartley guinea pigs weighing 200 to 300 g were killed by a blow to the head and the papillary muscles were rapidly excised from the right ventricles. The preparations were superfused with Tyrode solution. MEASUREMENTS AND MAIN RESULTS: The rate of onset of the use dependent inhibition of conduction velocity and that of the square of conduction velocity were both faster than the simultaneously measured rate of onset of dV/dtmax inhibition induced by 20 mumol.litre-1 quinidine at high frequency stimulation. The relation between the rates of onset of the use dependent inhibition of conduction velocity (and the square of conduction velocity) and dV/dtmax became weak with low frequency stimulation and in the presence of 60 mumol.litre-1 quinidine. However, the recovery of conduction velocity (and the square of conduction velocity) from quinidine induced use dependent blockade, as measured by the extrastimulation method, appeared to be slower than the recovery of dV/dtmax. These results may be explained by a transient change in intracellular and intercellular conditions, such as an increase in internal resistance. CONCLUSIONS: The onset and recovery of the use dependent inhibition of conduction by antiarrhythmic drug may not always parallel the changes of the dV/dtmax of action potential in multicellular muscle preparations.

Chronotropic effect of nizofenone fumarate in rabbit sino-atrial node in vitro.

1. The effects of nizofenone fumarate were studied on the membrane potentials and currents of rabbit sino-atrial node preparations by means of the double-microelectrode voltage clamp method. 2. In spontaneously firing pacemaker cells, nizofenone (above 1 microM) decreased the heart rate. Above 3 microM, nizofenone reduced the maximum upstroke velocity, the amplitude of the action potential and the slope of the phase 4 depolarization, and prolonged the action potential duration at 50% repolarization. 3. Under voltage clamp conditions, nizofenone decreased the slow inward current and the time-dependent potassium outward current in a dose-dependent manner. 4. These findings suggest that nizofenone exerts an inhibitory action on the automaticity of sinoatrial node preparations via effects on both inward and outward currents.

Mechanism of inhibition of the sodium current by bepridil in guinea-pig isolated ventricular cells.

1. Effects of bepridil, a sodium-, calcium-, and potassium-antagonistic agent, on the Na+ current were studied by the whole cell voltage clamp technique (tip resistance = 0.5 MOhm, [Na]i and [Na]o 10 mmol l-1 at 20 degrees C). 2. Bepridil produced tonic block (Kdrest = 295.44 mumol l-1, Kdi = 1.41 mumol l-1; n = 4). 3. Bepridil (100 mumol l-1) shifted the inactivation curve in the hyperpolarization direction by 13.4 +/- 2.7 mV (n = 4) without change in the slope factor. 4. In the presence of 50 mumol l-1 bepridil, bepridil showed use-dependent block at 2 Hz, whereas changes in pulse duration did not significantly effect this use-dependent block (81% +/- 2% at 10 ms, 84% +/- 3% at 30 ms, 86% +/- 3% at 100 ms; n = 4). 5. After removal of fast inactivation of the Na+ current by 3 mmol l-1 tosylchloramide sodium, bepridil (50 mumol l-1) still showed use-dependent block which was independent of the holding potential. 6. The recovery time constant from the bepridil-induced use-dependent block was 0.48 s at holding potential of -100 mV and 0.51 s at holding potential of -140 mV. 7. These results indicate that bepridil could bind to the receptor in the sodium channel through the hydrophobic and the hydrophilic pathway and leave the receptor through the hydrophobic pathway in the lipid bilayer. The binding and dissociation kinetics of this drug were shown to be fast, and the accumulation of the drug in the sodium channel appeared to be small. Bepridil is presumed to be safe in terms of adverse effects that result from drug-accumulation in the sodium channel.

The effect of TYB-3823, a new antiarrhythmic drug, on sodium current in isolated cardiac cells.

1. Sodium current (INa) blockade by TYB-3823, a newly synthesized antiarrhythmic agent, was investigated in isolated single ventricular myocytes by use of the whole cell patch-clamp technique. 2. TYB-3823 blocked INa under steady-state conditions (Kd,rest = 500 microM, Kd,i = 4.9 microM), findings consistent with a shift in the steady state INa availability curve to more negative potentials. 3. TYB-3823 produced use-dependent block at 2 Hz in conjunction with increase in pulse duration (5-300 ms), that was markedly enhanced at less negative holding potentials. 4. The time course of the onset of block was accelerated and the degree of use-dependent block was decreased at more negative holding potential. The time course of the onset of block was accentuated with enhancing block at more positive holding potentials. 5. The time course of recovery from use-dependent block was accelerated at more negative holding potentials but was accentuated at more positive holding potentials. 6. These results suggest that both tonic block and use-dependent block of sodium channels in cardiac tissue might result from an interaction of TYB-3832 with sodium channels mainly in the inactivated channel states and the kinetics of the interaction between drug and receptor may be modulated by the inactivation gate.

Effects of aprindine on conduction velocity and Vmax in guinea-pig papillary muscles.

One of the effects of antiarrhythmic drugs is the reduction of conduction velocity. Cable theory predicts that there is a nonlinear relationship between conduction velocity and upstroke velocity (Vmax) of action potential. By using conventional microelectrode techniques, aprindine-induced reduction of Vmax of action potential and conduction velocity in guinea-pig papillary muscles were measured. Aprindine-produced, use-dependent, and concentration-dependent changes in conduction velocity and the decline of square of conduction velocity was well fit by a single exponential. Time constants for square of conduction velocity were comparable to simultaneously measured time constants for effects of Vmax. At a concentration of 1 to 10 microM aprindine, onset changes between Vmax and conduction velocity had a log-linear relationship in a predicted fashion. Whereas, in the recovery process from aprindine-induced depression, slow recovery time course of conduction velocity was observed. In conclusion, in the presence of aprindine, only onset block of conduction velocity can be analyzed quantitatively in the relationship to observation on Vmax in vitro. These results suggested that in the presence of aprindine, the recovery of internal conductance may be slower than that of Vmax.

Amitriptyline inhibits the G protein and K+ channel in the cloned thyroid cell line.

We have reported that thyroid K+ channel is activated by extracellular application of the thyroid-stimulating hormone (TSH) using single channel recording method performed on cloned normal rat thyroid cell (FRTL-5) membrane. Treatment of dibutyryladenosine cyclic monophosphate (Bt2 cAMP) also activated the TSH-dependent K+ channel. These findings indicate that the thyroid K+ channel is activated through the TSH-adenosine cyclic monophosphate (cAMP)-protein kinase A system. We examined the effects of amitriptyline on TSH-guanosine triphosphate binding protein (G protein)-adenylate cyclase-cAMP-K+ channel system in the cloned normal rat thyroid cell line FRTL-5. Amitriptyline inhibited the cAMP production induced by TSH. Amitriptyline also inhibited the cAMP production induced by cholera toxin, indicating that amitriptyline inhibited the thyroid G protein. Amitriptyline had no effect on TSH-receptor binding and cAMP production by forskolin (adenylate cyclase stimulator). Amitriptyline inhibited the K+ channel activation by cAMP, indicating that the suppressing mechanism is not the inhibition of TSH receptor or G protein but the direct suppression of K+ channel. It was concluded that amitriptyline inhibited the thyroid G protein and K+ channel.

Evaluation of negative inotropic and antiarrhythmic effects of class 1 antiarrhythmic drugs.

The effects of 6 class 1 antiarrhythmic drugs (aprindine, cibenzoline, disopyramide, lidocaine, pirmenol, and quinidine) on myocardial action potential, its maximal upstroke velocity (Vmax) and isometric contractile force were evaluated by electrophysiological techniques. All the class 1 antiarrhythmic drugs examined had dose-dependent negative inotropic effects. The inhibitory effects on contractile force (Fc) was compared with the inhibitory effects on sodium channels under fast response. This ratio was indicated by IC50 Vmax/IC50Fc, and when evaluated and arranged in order of descending magnitude, these were the results: pirmenol, disopyramide, lidocaine, quinidine, cibenzoline and aprindine. The negative inotropic effects and the effects on action potential duration induced by these antiarrhythmic drugs were independent of each other; it had been found that classification of the drugs according to their effects on action potential duration did not provide sufficient information about negative inotropic effects. Class 1 antiarrhythmic drugs can be divided into 3 groups depending on the regression pattern of myocardial contractile force and the Vmax of action potentials under fast response and slow response. Drugs, whose inhibitory effects on sodium channels, are the main cause of negative inotropic effects. Drugs, whose inhibitory effects on calcium channels, are the main cause of negative inotropic effects. Drugs for which it is difficult to determine whether sodium channel or calcium channel blockade contributes more to their negative inotropic effects.

Actions of flunarizine dihydrochloride on the electrical activity of isolated guinea-pig papillary muscle and rabbit sino-atrial node.

Effects of flunarizine on electrophysiological properties of isolated guinea-pig papillary muscles and rabbit sino-atrial nodes were examined using conventional microelectrode and double-microelectrode voltage clamp methods. Although flunarizine did not affect the maximum rate of rise of the action potential (Vmax) in fast response fibers, the compound depressed the slow action potentials of K+-depolarized papillary muscle (at above 10 microM) and reduced the automaticity of sino-atrial node (at above 3 microM). In voltage clamp experiments, flunarizine selectively suppressed the slow inward current of sino-atrial node specimens. It is concluded that relatively high concentrations of flunarizine directly modify the electrical activity of isolated mammalian myocardium via a suppression of Ca2+ influx through the cell membrane.

Comparison of Ca2+ channel inhibitory effects of cibenzoline with verapamil on guinea-pig heart.

1. Cibenzoline, a class 1 antiarrhythmic drug, was studied on the effects upon the myocardial slow inward Ca2+ current (ICa) and the contractile force, and was compared with verapamil. 2. In voltage-clamp experiments with guinea-pig ventricular myocytes, cibenzoline caused a concentration-related inhibition of ICa (IC50 = 30 microM), whereas verapamil exerts a stronger effect (IC50 = 0.6 microM). 3. Cibenzoline also produced a concentration-related decrease in contractile force of isolated papillary muscle preparation (IC50 = 30 microM), while IC50 of verapamil was 0.8 microM. 4. Although the myocardial Ca2+ channel blocking effect of cibenzoline is about 1/50 compared to verapamil, cibenzoline possesses an inhibitory action on Ca2+ channel as well as on Na+ channel.

Quinidine enhances intracellular Ca2+ accumulation during rapid stimulation.

1. The quinidine-induced modification of intracellular Ca2+ concentration ([Ca2+]i) was studied in guinea-pig myocardium using fura-2. Quinidine reduced the systolic fluorescence signal level for [Ca2+]i and enhanced the end-diastolic signal level during a stimulation train. 2. The diastolic decay of [Ca2+]i fitted 2 exponential curves. Quinidine distorted the stimulation frequency-dependent acceleration of rapid [Ca2+]i decay, and prolonged the mean time constant of rapid decay after 2 Hz stimulation, from 154.4 to 205.3 msec (20 microM), and to 259.7 msec (60 microM quinidine). The time constant of slow recovery from [Ca2+]i accumulation after the stimulation train was not affected by stimulation frequency, or by quinidine, or caffeine. 3. These results suggest that quinidine modulates [Ca2+]i via a balance between the slowing of rapid [Ca2+]i decay and the reduction of the systolic [Ca2+]i. This effect may contribute to the anti-arrhythmic and pro-arrhythmic effects exerted by quinidine in some conditions.

[Clinical incidence of primary cardiac tumors].

Recently, clinical diagnostic methods for heart disease, especially echocardiography, have remarkably progressed and the incidence of cardiac tumors increased. This study investigated the characteristics of tumors in 115 patients with primary cardiac tumor, diagnosed histologically in 1993-1994, in collaboration with 126 university hospitals in Japan. Histological diagnosis, location, initial clinical manifestations and prognosis of cardiac tumors are reported. Of the 115 patients, 98 (85%) had primary benign tumor. Myxoma was the most common histological group (91 cases). Primary malignant cardiac tumors occurred in 17 patients (15%) of which rhabdomyosarcoma was the most frequent (5 cases). Sixty percent of patients with myxoma were female, a similar ratio to the proportion of female patients with other primary cardiac tumors. Most patients with myxoma had the tumor in the left side of the heart [76 cases (84%) in left side, 12 (13%) in right side, 3 (3%) in both sides]. In contrast, the primary malignant cardiac tumors occurred more in the right side of the heart rather than in the left side (eight in right side, six in left side). Embolization was the characteristic initial clinical manifestation for myxoma. There were no patients with initial manifestation of embolization in the other histological groups. Reflecting the recent progress and spread of clinical diagnostic methods, 17% of all patients were discovered asymptomatically. All patients with myxoma who were asymptomatic underwent operation, and there were no surgical deaths. In contrast, only one patient with primary malignant cardiac tumor could survive longer than a year. In conclusion, the progress and the spread of cardiovascular imaging has contributed to the early diagnosis of primary cardiac tumors. These data demonstrate new clinical and pathological characteristics of primary cardiac tumors seen in recent cardiologic practice in Japan.

Membrane actions of ethanol on rabbit sino-atrial node studied by voltage clamp method.

Electrophysiological effects of ethanol on rabbit sino-atrial node were studied by means of the double-microelectrode voltage clamp method. In spontaneously beating preparations, ethanol (above 1%) decreased the maximum rate of depolarization of the action potential. Furthermore, 3% ethanol produced a significant decrease in the action potential amplitude, the action potential duration at 50% repolarization and the rate of the diastolic depolarization. Of the current systems, voltage clamp studies showed that ethanol reduced the slow inward current, the time-dependent potassium outward current and the hyperpolarization activated inward current. These observations indicate that ethanol does not have an effect on a single current system, but directly modifies the spontaneous discharge of sino-atrial node pacemaker cells due to a reduction of the time-dependent current systems.

Effect of urapidil on the action potentials in the guinea-pig ventricular myocardium.

The electrophysiological effects of urapidil, a new alpha 1-adrenoceptor antagonist, were assessed in the reserpinized guinea-pig ventricular myocardium. Urapidil suppressed the maximal rate of rise (Vmax) of steady-state action potentials elicited by the fast responses at high concentrations independently of blockade of myocardial alpha-adrenoceptors, but not the Vmax of Ca(2+)-dependent slow action potentials of partially depolarized muscles in concentrations tested (up to 1.1 mM). Urapidil at high concentrations prolonged the action potential durations of the fast and slow responses in a manner similar to the quinidine-like antiarrhythmic drugs. These results suggest that the inhibitory effect of urapidil on the slow inward Ca2+ current and the Na+ current is in practice negligible.

Serum levels of lipids and apolipoproteins in angiographically assessed coronary artery disease in Japanese patients.

Serum total cholesterol (T-CHO), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and apolipoproteins (apo A-I, A-II, B, C-II, C-III and E) values were determined in 143 Japanese subjects undergoing coronary angiography. Among the factors measured, T-CHO, TG, apo B and C-III levels were significantly higher in patients with coronary artery disease (CAD) than in those without CAD. The HDL-C/T-CHO ratio was also significantly lower in patients with CAD. Although no parameters show differences between the group without CAD and the group of single vessel disease, T-CHO and apo B were significantly higher in the groups with double and triple vessel disease, and TG and apo C-III were also higher in the group with triple vessel disease compared with the normal group. Furthermore, the HDL-C/T-CHO ratio was significantly lower in the double and triple vessel groups, and HDL-C was lower only in the triple vessel group. The results indicate that changes in these parameters suggest a high likelihood of multiple vessel disease, and that an increase in TG and apo C-III levels is also one of the important indicators for CAD even in Japanese patients.

Effect of trapidil on serum lipid and apolipoprotein levels in patients with ischemic heart disease.

1. Serum total cholesterol (TCH), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein (LDL-C), atherosclerotic index (AI) and apolipoprotein (apo A-I, A-II, B, C-II, C-III and E) levels were investigated in patients with ischemic heart disease before and after medication of trapidil. 2. Twenty-one patients were orally given 100 mg of trapidil, three times daily (300 mg/day). After 8 weeks' administration, serum HDL-C level increased (P < 0.01) and AI decreased (P < 0.02) significantly, whereas TCH, TG and LDL-C levels tended to decrease but not significantly. 3. Among the parameters of apolipoproteins, apo A-I, a main protein of HDL-C, was significantly increased (P < 0.05) by trapidil. 4. These results indicate that trapidil has a beneficial effect on the coronary risk profile as reflected by lipid measurements.