Kinetics of rate-dependent shortening of action potential duration in guinea-pig ventricle; effects of IK1 and IKr blockade.

1. The kinetics of shortening of action potential duration (APD) following an increase in pacing rate, from 2 to 3.3 Hz, was characterized in guinea-pig ventricular preparations. Terikalant (RP62719), an inhibitor of the inwardly rectifying K+ current (IK1), and dofetilide, a specific inhibitor of the rapidly activating delayed-rectifier current (IKr), were applied to determine the effect of inhibition of these ion currents on slow APD shortening. 2. Action potentials were recorded from isolated guinea-pig ventricular myocytes using the perforated-patch patch-clamp technique, and monophasic action potentials were recorded from Langendorff-perfused guinea-pig ventricles using a contact epicardial probe. 3. Under control conditions, after an increase in pacing rate, APD immediately decreased, and then shortened slowly with an exponential time course. In ventricular myocytes, the time constant of this exponential shortening was 28+/-4 s and the amount of slow shortening was 21.9+/-0.9 ms (n=8) for an increase in rate from 2 to 3.3 Hz. Similar values were observed in Langendorff-perfused ventricles. 4. Terikalant dose-dependently increased APD and the increase was enhanced by rapid pacing ('positive' rate-dependence). The drug dose-dependently decreased the time constant of shortening and amount of slow APD shortening. In contrast, dofetilide, an inhibitor of IKr, which shows 'reverse' rate-dependent APD widening, had no significant effect on the time constant or amount of slow shortening. 5. These observations suggest that IK1 plays a role in rate-dependent shortening of APD. The results appear to support the hypothesis that 'reverse' rate-dependent effects of IKr blockers are due to these drugs not affecting the ion current(s) mediating intrinsic rate-dependent slow shortening of APD.

Dofetilide enhances shock-induced extension of refractoriness and lowers defibrillation threshold.

OBJECTIVE: To explore the hypothesis that class III antiarrhythmics reduce defibrillation threshold (DFT) by prolonging postshock refractoriness. ANIMALS AND METHODS: The effect of a new selective potassium channel blocker, dofetilide (DOFlow 2.5 micrograms/kg bolus plus 0.9 microgram/kg/h; DOFmed 10 micrograms/kg bolus plus 3.6 micrograms/kg/h; and DOFhigh 25 micrograms/kg bolus plus 9 micrograms/kg/h), on DFT was compared with that of placebo in anesthetized open chest dogs (n = 6 per group). The effects of dofetilide on refractory period extension (RPE) were assessed by using DFT strength shocks delivered at various stages of repolarization. RESULTS: DFT was significantly decreased in the DOFhigh group, whether expressed as shock peak voltage or energy (P < 0.05 compared with changes in placebo). At baseline, a shock timing of ventricular effective refractory period of 25 ms resulted in RPE of 100 +/- 24 ms, 80 +/- 11 ms, 91 +/- 14 ms and 90 +/- 20 ms in the placebo, DOFlow, DOFmed, and DOFhigh groups, respectively. After infusion, these RPE values were unchanged in the placebo group but tended to increase in the dofetilide treatment groups. DOFhigh significantly increased RPE by 20 +/- 18 ms (P < 0.05 compared with baseline values and changes in placebo). Dofetilide-induced changes in RPE and DFT were significantly correlated when expressed as voltage (r2 = 0.78, P < 0.01), current (r2 = 0.80, P < 0.01) and energy (r2 = 0.53, P < 0.01). CONCLUSIONS: These results show that dofetilide prolonged RPE at a plasma level that reduced DFT, thus providing support for the hypothesis that selective prolongation of refractoriness may synergize with shock-induced RPE to decrease the energy requirements for defibrillation.

Development and evaluation of a fully automated monophasic action potential analysis program.

A fully automated program has been developed for analysing digitised cardiac monophasic action potentials (MAPs) of several animal species. Under diverse conditions, the program's performance is evaluated by comparison with high-resolution manual analysis of action potential duration, MAP detection, baseline and plateau determination. For each variable, the differences between both forms of analysis are categorised according to species (rabbit, dog or baboon), intervention (control, sematilide or LY-190147), rhythm source (sinus or paced), and rates. They are then examined using ANOVA (null hypothesis: no difference between methods for any category). Intra-group differences are significant for baseline selection (p < 0.05), and treatment effects are significant (p < 0.05) for one species (baboon). However, the differences found are negligible when compared with expected measurement errors. Thus the program reliably detects and accurately analyses MAPs, independent of species, drug or pacing. The program's strength is its use of simple heuristic algorithms to identify and profile MAP signals while rejecting spurious transients. These algorithms focus on the procedural aspects of automatic MAP detection and validation, with minimum use of complex mathematics. The program's estimated throughput exceeds 8200 MAPs min-1 on a 50 MHz 1486 DX machine. With available data format conversion packages, signals from almost any data-acquisition source can be analysed.

Class III antiarrhythmic effects of LY-190147 on defibrillation threshold.

Defibrillation strength shocks delivered within an action potential (AP) delay repolarization. Shock-induced AP duration extension (APDE) may prolong refractoriness and terminate or prevent reinitiation of reentry, favoring defibrillation. This study examined LY-190147 (LY) effects on defibrillation threshold (DFT) in 11 dogs. Ventricular effective refractory period (VERP) and epicardial monophasic AP duration at 75% repolarization (APD75) were recorded at 300-, 400-, 500-, and 600-ms pacing cycle length (CL). APDE was measured as the time to 50% repolarization after a DFT strength shock delivered at 50, 25, and 0 ms before or 25 ms after VERP during pacing at 300 ms CL in 4 of the dogs. We made all recordings before drug administration and after infusions of 0.03, 0.3, and 3.0 mg/kg LY, using 1.5-h dosing intervals. LY lowered DFT in a saturating dose-response manner whether expressed as shock peak voltage (V) or energy. LY decreased DFT-V from 357 +/- 77 V before drug to 331 +/- 60 V (-6 +/- 12%), 290 +/- 43 V (-17 +/- 13%, p < 0.001), and 312 +/- 45 V (-11 +/- 12%, p < 0.05) at 0.03, 0.3, and 3.0 mg/kg, respectively. Similarly, LY treatment decreased defibrillation energy requirements from 6.9 +/- 2.7 J before drug by 7 +/- 25%, 26 +/- 24%, and 12 +/- 25% at the same doses. At 300-600 ms CL, LY prolonged APD75 by an average of 10 +/- 8% at 0.03 mg/kg, 17 +/- 6% at 0.3 mg/kg, and 24 +/- 9% at 3 mg/kg. At these CL, LY prolonged VERP by an average of 4 +/- 6% at 0.03 mg/kg, 15 +/- 10% at 0.3 mg/kg, and 11 +/- 9% at 3 mg/kg. APDE was increased from 62 +/- 9 ms before to 68 +/- 14, 80 +/- 16 (p < 0.001) and 72 +/- 13 ms (p < 0.05) at 0.03, 0.3, and 3.0 mg/kg LY, respectively. Therefore, LY prolonged VERP and APDE and affected DFT in the same saturating dose-response manner. LY may facilitate defibrillation by increasing the duration of postshock refractoriness.

Effects of optical enantiomers CK-4000(S) and CK-4001(R) on defibrillation and enhancement of shock-induced extension of action potential duration.

INTRODUCTION: Class III antiarrhythmics have been reported to lower defibrillation threshold (DFT); however, the mechanism(s) of this effect is unknown. Recent evidence suggests that DFT strength DC shocks may terminate reentrant arrhythmias through prolongation of action potential duration and refractory periods. Since Class III antiarrhythmic drugs prolong repolarization, we examined the hypothesis that these drugs enhance shock-induced action potential duration extension (APDE), which might contribute to lowering of DFT. METHODS AND RESULTS: In order to investigate the specificity of drug effects on action potential repolarization following a shock, an optical enantiomer with mixed beta-blocking and Class III effects (CK-4000) and its enantiomer with "pure" Class III antiarrhythmic effects (CK-4001) were compared against placebo in a canine defibrillation model (n = 8 per group). At the 3 mg/kg dose, the enantiomers nonstereoselectively lowered DFT voltage by 19 +/- 15% (CK-4000, P < 0.05 compared to placebo) and 25 +/- 12% (CK-4001, P < 0.05 compared to placebo), indicating that Class III antiarrhythmic actions alone were sufficient for this effect. Similarly, CK-4000 and CK-4001 at 3 mg/kg enhanced APDE (P < 0.01 compared to placebo) by 20 +/- 11% and 24 +/- 17%, respectively. APDE prolongation significantly correlated with reduction in DFT voltage for both CK-4000 (r = -0.55, P < 0.03) and CK4001 (r = -0.63, P < 0.01). At 3 mg/kg, the enantiomers stereoselectively prolonged action potential duration (APD75) by an average of 37 +/- 14% (CK-4000, P < 0.001) and 23 +/- 14% (CK-4001, P < 0.001), and ventricular effective refractory period (VERP) by 38 +/- 15% (CK-4000, P < 0.01) and 27 +/- 13% (CK-4001, P < 0.05). Prolongations of APD75 and VERP did not correlate with reductions of DFT in individual dogs. CONCLUSIONS: These results show that Class III antiarrhythmics and DFT strength shocks additively delay repolarization, which suggests that drug enhancement of APDE may contribute to their effects on DFT.