Ventricular Monophasic Action Potentials Research Articles

Abstract 11651: Machine Learning of Action Potential Shape to Define Refractory Periods in Ischemiccardiomyopathy

Introduction: The effective refractory period (ERP) can characterize arrhythmic risk. To measure it clinically, a pacing protocol with 8-10 beats is required, preventing beat to beat analysis. Monophasic action potentials (MAP) can indicate ERP on every single beat (Fig A), but are often difficult to annotate due to a wandering baseline (Fig B). Hypothesis: We hypothesized that machine learning of ventricular MAPs could help identify instantaneous repolarization, without requiring manual measurement and without measuring time to drop to a specific repolarization voltage. Methods: We analyzed 5706 MAPs from 42 patients (age 65±13 Y) with ischemic cardiomyopathy in pacing. A multilayer perceptron (MLP) was developed to input MAP shapes and output action potential duration at 30, 60, 90% (MAPD 30,60,90) repolarization, indicating ERP. We used 60% of data in training, 20% in validation, 20% in testing. Results were compared to manual measurement. To identify which MAP regions were key to machine learning, we systematically perturbed MAP regions with white noise and assessed the drop in performance. Results: Errors between APD 30,60 and 90 from MLP and manual marking were 9.3(2.0), 5.8 (1.1) and 5.8 (0.7) ms respectively (fig. C). Fig D shows that MAP phases 0 and 3 were used by MLP to predict APD, as seen in the significant increase in error with perturbations in those MAP regions ( P <0.05). Conclusions: Machine learning can directly indicate effective refractory periods by learning the electrogram shape of single beats, without extrastimuli or manual measurement. Explainability studies confirm that machines learn physiological data. This approach can be used to assess beat-to-beat characterization of atrial fibrillation patients.

Reduction of artifacts and noise in small electrogram datasets without manual annotation using transfer machine learning

Abstract Background/Introduction Mapping AF is challenging. Monophasic action potentials (MAPs) show that most of the recorded signals are not representing the atrial activity, and arise from far-field or other artifacts. Thus, reducing noise in electrophysiological signals is essential, yet it can be difficult for cross-talk from other chambers and pacing. Strategies to reduce noise include template matching, averaging, and smoothing, but all of them have major limitations. Furthermore, expert interpretation requires knowledge to discriminate signals from noise, but is subjective. Purpose We hypothesised a) that atrial and ventricular electrograms with varying artifact and noise can be denoised using autoencoder neural networks (NNs) without requiring manual annotation and in a reproducible manner, and b) we could train these NNs on a large available dataset ventricular signals, then apply transfer learning to the original smaller atrial dataset. We applied this approach to MAPs, which have interpretable shapes and would help identifying local from far-field signals helping in diagnosis, mapping and ablation. Methods We first trained with 5706 left and right ventricular MAPs from 42 patients with ischemic cardiomyopathy (age 65±13y; Fig. 1A), with 60% for training, 20% (validation) and 20% (testing). Transfer learning and parameter-tuning were then used to apply this NN to a smaller sample of atrial MAPs (N=641 from 21 patients, 67±5y, 13 women; Fig. 2B, D, F). The autoencoder was used to eliminate pacing artifacts in ventricular MAPs (Fig. 1B, C) and denoise atrial MAPs (Fig. 2C, E, G) by reconstructing key learned features. The accuracy of the reconstruction was evaluated using Pearson Correlation Coefficient (PCC) and a novel similarity coefficient (SC). No manual annotation was required to identify noisy signals. Results The trained NN encoder learned key features of ventricular MAPs and reconstructed these clean signals with a SC=0.91±0.16 and PCC=0.99±0.01 (Fig. 1A). With this training, the NN was able to denoise ventricular MAPs with pacing artifact (Fig. 1B, C). After fine-tuning, the NN learned key signal features (upstroke, triangular descent, terminus) and thus reduced diverse noise without specific training or manual annotation. Namely, it was able to reconstruct atrial MAPs eliminating ventricular noise, high frequency noise and truncated signals (Fig. 2). Conclusions Machine learned encoder-decoders are powerful tools that can learn essential features of atrial and ventricular signals and hence isolate noise. Transfer learning is effective when large datasets are unavailable for training. This approach can separate atrial beats from far-field ventricular beats and other sources of noise. The ability to eliminate a diverse range of noise improves this approach over existing techniques and may have far-reaching applications in electrophysiology, such as mapping and ablation. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): NIH

Artificial intelligence to reduce artifact in cardiac electrophysiological signals

Abstract Background/Introduction Signals in Electrophysiology cases are often noisy despite laboratory shielding and filtering, and current noise-reduction methods are suboptimal. Template matching can identify a “nearest type” of electrogram, but libraries of signal shapes may be unavailable. Beat averaging can reduce noise but obscures beat-to-beat variations and is not optimal to analyze dynamically changing signals, such as when moving a catheter in the heart. Smoothing reduces noise yet blurs high frequency components. Purpose We set out to test if machine learned autoencoders could reduce noise in single beats without requiring massive training data or beat libraries. Specifically, we hypothesised that noisy electrograms in small datasets of atrial signals could be de-noised using an encoder-decoder neural network (NN) using transfer learning of machines trained to recognize key features in larger datasets of related signals. Methods We applied NN to monophasic action potentials (MAPs), because they have visually verifiable shapes. The NN was first trained to reconstruct 5706 left and right ventricular MAPs in 42 patients (67±13y; Fig. 1A). Transfer learning was then used to apply the NN to a much smaller dataset of 641 atrial MAPs in 21 patients (67±5y, 13 women; Fig. 1B, D, F). Results NN reconstructed atrial MAPs with a Pearson correlation of 0.87±0.11. After fine-tuning, NN reconstruction accuracy improved dramatically (Pearson 0.99±0.01; p<0.001). In Fig. 1B–G the NN learned key MAP features (upstroke, triangular descent, terminus) and thus could eliminate ventricular artifact and electrical circuit noise without specific training or manual annotation. Conclusion Machine learned autoencoders are a novel and powerful approach to de-noise electrophysiological signals in a dynamic, beat-to-beat fashion. The ability to learn fundamental signal features from models trained in large datasets, and apply them via transfer learning to small datasets in different heart chambers may have wide ranging applications for automated signal annotation, mapping and ablation. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): NIH

Defining refractoriness in single atrial beats using autoencoder neural networks

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): NIH Background Mapping atrial fibrillation (AF) is complicated by signals which may be local or far-field, but which cannot currently be separated. This could be clarified by a knowledge of atrial refractory periods, yet these are difficult to define from monophasic action potentials (MAP) in patients. We hypothesized that transfer learning using an autoencoder neural network (ANN), first trained with less-noisy ventricular signals, can be applied to de-noise and classify atrial MAPs. Methods We first developed an ANN to encode MAPs in 5706 ventricular MAPs from N=42 patients (age 65±13y) during pacing (fig1. A-B). This created a latent feature space. We now tuned the ANN to classify atrial MAPs in a different cohort of patients with AF. We used a statistical loss function based on mathematical optimization to evaluate the accuracy of final representations of the MAP and classify the different signals. Results The autoencoder ANN reconstructed ventricular MAPs with an average similarity of 0.85 (range 0-1) (an example is shown in fig 1.B). We tested on 3000 atrial MAPs in AF patients (N=21; 67±5y, 13 women). Atrial MAPs were accurately represented (fig 1.E-F) with similarity indices that were higher than those obtained by a panel of 3 experts. Fig. 1 shows the reconstruction of different signals: ventricular MAP (fig 1.A-B), ventricular MAP with pacing artifact (fig. 1.C-D), atrial MAP (transfer learning is assumed in here; fig 1.E-F) and noise or signals with morphologies of no interest (fig 1.G-H). Fig. 2 shows the classification of signals according to the similarity metric that allows distinguishing among the different types of signals without manual annotation (p<0.05 between groups). Conclusion Atrial refractory periods can be defined in single beats in AF patients using autoencoder neural networks and transfer learning. This approach can separate atrial beats from far-field ventricular beats and other sources of noise. Future work can study if this approach can be used to improve AF mapping or define novel physiological phenotypes.

Noise reduction in electrophysiological signals using transfer machine learning

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): NIH Background/Introduction Reducing electrophysiological signal noise is essential for diagnosis, mapping and ablation, yet most approaches are suboptimal. Template matching requires libraries of known signal types, that are difficult to obtain. Beat averaging can reduce noise, yet cannot be applied to single beats and obscures beat-to-beat variations. Beat smoothing can lose critical and subtle signal features. We set out to use neural networks (NN) based on encoder-decoders, which are able to extract key signal features and hence reconstruct them without noise and artifact. Purpose We hypothesised that electrograms with varying sources of artifact can be denoised using autoencoder neural networks. We further hypothesised that this could be achieved in a small data set by developing the method in a larger dataset of related signals, then using transfer learning. We tested this approach for atrial monophasic action potentials (MAPs) that have verifiable shapes. Methods The NN was first trained with 5706 left and right ventricular MAPs from 42 patients with ischemic cardiomyopathy (age 65±13y; fig 1.A): 60% for training, 20% (validation) and 20% (testing). Transfer learning and parameter-tuning were then used to apply this NN to a smaller sample of atrial MAPs (N=641, 21 patients, 67±5y, 13 women; fig D,F,H). Results The autoencoder was able to learn key features of MAPs, and hence reconstruct them without artifacts. NN learned ventricular MAPs with similarity coefficient 0.91±0.16, Pearson correlation 0.99± 0.01 (fig A) and learned key features (upstroke, triangular descent, terminus) to reduce noise (fig B-C). Applying this trained NN to atrial MAPs, the approach automatically eliminated ventricular artifact (fig E), high frequency noise (fig G), truncation (fig I), saturation and other artifacts. After fine-tuning, the NN reconstructed atrial MAPs with Pearson correlation = 0.99±0.01 (p<0.001). Conclusions Machine learned encoder-decoders are powerful tools that can automatically eliminate diverse types of noise in single beats by learning essential signal features. Transfer learning makes this possible without large datasets for training, even from signals in a different cardiac chamber. This approach may have far-reaching applications for mapping and ablation.

Effects of post-treatment electroacupuncture on ventricular monophasic action potential and cardiac function in a rat model of ischemia/reperfusion injury.

To determine the effects of post-treatment electroacupuncture (EA) on the electrophysiological properties of ventricular muscle in rats with ischemia/reperfusion (IR) injury. Male Sprague-Dawley (SD) rats were randomly assigned into sham-operated (SH), IR and IR + EA groups (n = 8 each). The IR model was generated by ligation of the left anterior descending (LAD) coronary artery for 30 min. After establishing the IR model, EA was administered at PC6 for 30 min while opening the coronary artery and allowing reperfusion for 30 min. Heart rate (HR), mean arterial pressure and monophasic action potential (MAP) of cardiac muscle in the outer membrane of the antetheca of the left ventricle before coronary artery ligation (T0), after coronary artery ligation for 30 min (T1) and after reperfusion for 30 min (T2) were recorded. At the same time, ventricular electrophysiological parameters including ventricular effective refractory period (ERP), conduction velocity (CV) and ventricular fibrillation threshold (VFT) were measured. Then, the cardiac function and the levels of creatine kinase-muscle/brain (CK-MB) and cardiac troponin I (cTnI) were monitored. Based on these data, monophasic action potential amplitude (MAPA), the maximum depolarization velocity (Vmax) and the MAP durations at 50% and 90% repolarization (MAPD50 and MAPD90) were calculated to determine the incidence of arrhythmia during reperfusion. Compared with the SH group, the IR group showed an obviously decreased HR as well as reduced mean arterial pressure, Vmax, CV, ERP and MAPA. All indices of cardiac function except left ventricular end-diastolic pressure (LVEDP) decreased (i.e. ventricular systolic pressure (LVSP), left ventricular ejection fraction (LVEF), fractional shortening (FS) and rate of the ventricular pressure rise/drop (±dp/dt)). Furthermore, the MAPD50 and MAPD90 were prolonged, and the levels of CK-MB and cTnI increased (p < 0.05). In comparison to the IR group, HR and the mean arterial pressure were increased. All indices of cardiac function except LVEDP increased (LVSP, LVEF, FS and ±dp/dt). Vmax, CV, ERP and MAPA were also increased in the IR + EA group. However, MAPD50 and MAPD90 were distinctly shortened, and the levels of CK-MB and cTnI decreased (p < 0.05). There were no statistically significant differences in VFT between the three groups (p > 0.05). EA post-treatment can relieve prolongation of repolarization and slowed depolarization of ventricular muscle during IR, thus decreasing the rate of incidence of reperfusion arrhythmia.

Prolonged QT intervals in mice with cardiomyocyte-specific deficiency of the molecular clock.

Cardiac arrhythmias and sudden deaths have diurnal rhythms in humans. The underlying mechanisms are unknown. Mice with cardiomyocyte-specific disruption of the molecular clock genes have lower heart rate than control. Because changes in the QT interval on the electrocardiogram is a clinically used marker of risk of arrhythmias, we sought to test if the biological rhythms of QT intervals are dependent on heart rate and if this dependency is changed when the molecular clock is disrupted. We implanted radio transmitters in male mice with cardiomyocyte-specific Bmal1 knockout (CBK) and in control mice and recorded 24-h ECGs under diurnal and circadian conditions. We obtained left ventricular monophasic action potentials during pacing in hearts ex vivo. Both RR and QT intervals were longer in conscious CBK than control mice (RR: 117±7 vs 110±9ms, P<.05; and QT: 53±4 vs 48±2ms, P<.05). The prolonged QT interval was independent of the slow heart rate in CBK mice. The QT interval exhibited diurnal and circadian rhythms in both CBK and control mice. The action potential duration was longer in CBK than in control mice, indicating slower repolarization. Action potential alternans occurred at lower pacing rate in hearts from CBK than control mice (12±3 vs 16±2Hz, respectively, P<.05). The bradycardic CBK mice have prolonged ventricular repolarization independent of the heart rate. Diurnal and circadian rhythms in repolarization are preserved in CBK mice and are not a consequence of the 24-h rhythm in heart rate. Arrhythmia vulnerability appears to be increased when the cardiac clock is disrupted.

Measures of repolarization variability predict ventricular arrhythmogenesis in heptanol-treated Langendorff-perfused mouse hearts.

BackgroundTime-domain and non-linear methods can be used to quantify beat-to-beat repolarization variability but whether measures of repolarization variability can predict ventricular arrhythmogenesis in mice have never been explored.MethodsLeft ventricular monophasic action potentials (MAPs) were recorded during constant right ventricular 8 ​Hz pacing in Langendorff-perfused mouse hearts, in the presence or absence of the gap junction and sodium channel inhibitor heptanol (0.1, 0.5, 1 or 2 ​mM).ResultsUnder control conditions, mean action potential duration (APD) was 39.4 ​± ​8.1 ​ms. Standard deviation (SD) of APDs was 0.3 ​± ​0.2 ​ms, coefficient of variation was 0.9 ​± ​0.8% and the root mean square (RMS) of successive differences in APDs was 0.15 ​± ​0.14 ​ms. Poincaré plots of APDn+1 against APDn revealed ellipsoid morphologies with a SD along the line-of-identity (SD2) to SD perpendicular to the line-of-identity (SD1) ratio of 4.6 ​± ​2.1. Approximate and sample entropy were 0.61 ​± ​0.12 and 0.76 ​± ​0.26, respectively. Detrended fluctuation analysis revealed short- and long-term fluctuation slopes of 1.49 ​± ​0.27 and 0.81 ​± ​0.36, respectively. Heptanol at 2 ​mM induced ventricular tachycardia in five out of six hearts. None of the above parameters were altered by heptanol during which reproducible electrical activity was observed (KW-ANOVA, P ​> ​0.05). Contrastingly, SD2/SD1 decreased to 2.03 ​± ​0.41, approximate and sample entropy increased to 0.82 ​± ​0.12 and 1.45 ​± ​0.34, and short-term fluctuation slope decreased to 0.82 ​± ​0.19 during the 20-s period preceding spontaneous ventricular tachy-arrhythmias (n ​= ​6, KW-ANOVA, P ​< ​0.05).ConclusionMeasures of repolarization variability, such as SD2/SD1, entropy, and fluctuation slope are altered preceding the occurrence of ventricular arrhythmogenesis in mouse hearts. Changes in these variables may allow detection of impending arrhythmias for early intervention.

Machine Learned Cellular Phenotypes in Cardiomyopathy Predict Sudden Death.

Susceptibility to VT/VF (ventricular tachycardia/fibrillation) is difficult to predict in patients with ischemic cardiomyopathy either by clinical tools or by attempting to translate cellular mechanisms to the bedside. To develop computational phenotypes of patients with ischemic cardiomyopathy, by training then interpreting machine learning of ventricular monophasic action potentials (MAPs) to reveal phenotypes that predict long-term outcomes. We recorded 5706 ventricular MAPs in 42 patients with coronary artery disease and left ventricular ejection fraction ≤40% during steady-state pacing. Patients were randomly allocated to independent training and testing cohorts in a 70:30 ratio, repeated K=10-fold. Support vector machines and convolutional neural networks were trained to 2 end points: (1) sustained VT/VF or (2) mortality at 3 years. Support vector machines provided superior classification. For patient-level predictions, we computed personalized MAP scores as the proportion of MAP beats predicting each end point. Patient-level predictions in independent test cohorts yielded c-statistics of 0.90 for sustained VT/VF (95% CI, 0.76-1.00) and 0.91 for mortality (95% CI, 0.83-1.00) and were the most significant multivariate predictors. Interpreting trained support vector machine revealed MAP morphologies that, using in silico modeling, revealed higher L-type calcium current or sodium-calcium exchanger as predominant phenotypes for VT/VF. Machine learning of action potential recordings in patients revealed novel phenotypes for long-term outcomes in ischemic cardiomyopathy. Such computational phenotypes provide an approach which may reveal cellular mechanisms for clinical outcomes and could be applied to other conditions.

Quantification of Beat-To-Beat Variability of Action Potential Durations in Langendorff-Perfused Mouse Hearts.

Background: Beat-to-beat variability in action potential duration (APD) is an intrinsic property of cardiac tissue and is altered in pro-arrhythmic states. However, it has never been examined in mice.Methods: Left atrial or ventricular monophasic action potentials (MAPs) were recorded from Langendorff-perfused mouse hearts during regular 8 Hz pacing. Time-domain, frequency-domain and non-linear analyses were used to quantify APD variability.Results: Mean atrial APD (90% repolarization) was 23.5 ± 6.3 ms and standard deviation (SD) was 0.9 ± 0.5 ms (n = 6 hearts). Coefficient of variation (CoV) was 4.0 ± 1.9% and root mean square (RMS) of successive differences in APDs was 0.3 ± 0.2 ms. The peaks for low- and high-frequency were 0.7 ± 0.5 and 2.7 ± 0.9 Hz, respectively, with percentage powers of 39.0 ± 20.5 and 59.3 ± 22.9%. Poincaré plots of APDn+1 against APDn revealed ellipsoid shapes. The ratio of the SD along the line-of-identity (SD2) to the SD perpendicular to the line-of-identity (SD1) was 8.28 ± 4.78. Approximate and sample entropy were 0.57 ± 0.12 and 0.57 ± 0.15, respectively. Detrended fluctuation analysis revealed short- and long-term fluctuation slopes of 1.80 ± 0.15 and 0.85 ± 0.29, respectively. When compared to atrial APDs, ventricular APDs were longer (ANOVA, P < 0.05), showed lower mean SD and CoV but similar RMS of successive differences in APDs and showed lower SD2 (P < 0.05). No difference in the remaining parameters was observed.Conclusion: Beat-to-beat variability in APD is observed in mouse hearts during regular pacing. Atrial MAPs showed greater degree of variability than ventricular MAPs. Non-linear techniques offer further insights on short-term and long-term variability and signal complexity.

Measurement variability of right atrial and ventricular monophasic action potential and refractory period measurements in the standing non-sedated horse

BackgroundIn human and veterinary medicine, monophasic action potential (MAP) analysis and determination of local refractory periods by contact electrode technique gives valuable information about local cardiac electrophysiological properties. It is used to investigate dysrhythmias and the impact of drugs on the myocardium. Precise measurement of total MAP duration is difficult, therefore the MAP duration is usually determined at a repolarization level of 90% (APD90). Until now, no studies are published about the feasibility of this technique in the standing non-sedated horse. In 6 healthy Warmblood horses, on two different days, an 8F quadripolar contact catheter was passed through a jugular introducer sheath and placed under ultrasound guidance at the level of the intervenous tubercle or right atrial free wall (RA), and in the right ventricular apex (RV) to record the MAP. The MAP amplitude and APD90 were measured at a resting sinus rhythm (heart rate of 30-42 bpm) and at pacing cycle lengths (PCL) of 1000 and 600 ms. The effective refractory period (ERP) was determined at PCL of 1000 and 600 ms.ResultsThe overall mean (±SD) APD90 (rest), APD90 (1000) and APD90 (600) were 263 ± 39 ms, 262 ± 41 ms, 236 ± 47 ms for the RA and 467 ± 23 ms, 412 ± 38 ms, 322 ± 29 ms for the RV. The mean ERP1000 and ERP600 were 273 ± 24 ms and 256 ± 22 ms for the RA and 386 ± 40 ms and 293 ± 30 ms for the RV. The measurement variability for the amplitude, APD90 and ERP measurements in the RA ranged between 36 and 44, 9–22 and 7–8%, respectively. The measurement variability for the amplitude, APD90 and ERP measurements in the RV ranged between 49 and 66, 6–7 and 10–12%, respectively.ConclusionsRA and RV MAP duration and ERP can be obtained by a contact electrode in standing non-sedated horses. The measurement variability varies with catheter location.

Pharmacokinetics and electrophysiological effects of sotalol hydrochloride in horses.

Arrhythmias in horses may require long-term anti-arrhythmic therapy. Unfortunately, oral anti-arrhythmic drugs for use in horses are currently scarce. In human patients and small animals, sotalol, a β-blocker with class III anti-arrhythmic properties, is often used for long-term treatment. To determine the pharmacokinetics of sotalol at multiple oral dosages in unfasted horses, as well as the effects on electro- and echocardiographic measurements, right atrial and ventricular monophasic action potential (MAP) and effective refractory period (ERP). Placebo controlled, double-blinded experiment. Six healthy, unfasted Warmblood horses were given either 0, 2, 3 or 4 mg/kg bodyweight (bwt) sotalol orally (PO) twice daily (bid) for 9 days in a randomised cross-over design. Echocardiography and surface electrocardiography were performed and plasma concentrations of sotalol and right atrial and right ventricular MAPs and ERPs were determined at steady-state conditions. Statistical analysis was performed using a repeated measures univariate analysis with post hoc Bonferroni corrections. Calculated mean steady-state plasma concentrations determined by nonlinear mixed-effect modelling were 287 (range 234-339), 409 (359-458) and 543 (439-646) ng/mL for 2, 3 and 4 mg/kg bwt sotalol PO bid respectively. Sotalol significantly increased the QT interval and ERPs, but, despite increasing plasma concentrations, higher dosages did not result in a progressive increase in QT interval or ERPs. Echocardiographic and other electrocardiographic measurements did not change significantly. MAP durations at 90% repolarisation were not significantly different during sotalol treatment. Besides transient local sweating, no side effects were noted. Study size and ad libitum feeding of hay. Sotalol at a dose of 2, 3 and 4 mg/kg bwt PO bid increases the QT interval and ERP and might be a useful drug for long-term anti-arrhythmic therapy in horses.

Deleterious acute and chronic effects of bradycardic right ventricular apex pacing: consequences for arrhythmic outcome.

In the chronic complete atrioventricular (AV) block dog (CAVB) model, both bradycardia and altered ventricular activation due to the uncontrolled idioventricular rhythm contribute to ventricular remodeling and the enhanced susceptibility to Torsade de Pointes (TdP) arrhythmias. We investigated the effect of permanent bradycardic right ventricular apex (RVA) pacing on mechanical and electrical remodeling and TdP. In 23 anesthetized dogs, serial experiments were performed at sinus rhythm (SR), acutely after AV block (AAVB) and 3weeks of remodeling CAVB at a fixed pacing rate of 60/min. ECG, and left (LV) and right ventricular (RV) monophasic action potentials durations (MAPD) were recorded; activation time (AT) and activation recovery interval (ARI) were determined from ten distinct LV electrograms; interventricular mechanical delay (IVMD) and time-to-peak strain (TTP) of the LV septal and lateral wall (ΔTTP: lateral wall minus septal wall) were obtained echocardiographically. Dofetilide (25μg/kg/5min) was infused to study TdP inducibility. In baseline AAVB, in comparison to SR, RVA bradypacing acutely increased QT interval, LV, and RVMAPD. Echocardiographic IVMD and ΔTTP were initially increased, which was partially corrected after 3weeks of RVA pacing (IVMD: 22±13 vs. 42±11 vs. 31±6ms; ΔTTP: -2±47 vs. -114±38 vs. -36±22ms). QT interval (362±23 vs. 373±29ms), LVMAPD (245±18 vs. 253±22ms), RVMAPD (226±26 vs. 238±31ms), and mean LV-ARI (268±5 vs. 267±6ms) were not significantly changed after 3weeks of RVA pacing. During AAVB, dofetilide increased mean LV-ARI (381±11ms) with largest increases in the later activated basal areas (slope AT-ARI: +0.96). In contrast with acute RVA pacing, 3week pacing increased TdP inducibility (0/13 vs. 11/21) and mean LV-ARI (484±18ms), while the slope of AT-ARI responded differently on dofetilide (-2.37), with larger APD increases in the early region. The latter was supported at the molecular level: reduced RNA expressions of three repolarization-related ion channel genes in early (KCNQ1, KCNH2, and KCNJ2) versus two in late regions (KNCQ1 and KCNJ2). In conclusion, bradycardic RVA pacing acutely induced LV intra- and interventricular mechanical dyssynchrony, which was partially reversed after 3weeks of pacing (remodeling). The latter occurred without apparent baseline electrical effects. However, dofetilide clearly unmasked (region-specific) arrhythmic consequences of remodeling.

Abstract 19945: Acute Alcohol Intoxication and Hemorrhagic Shock Lead to Microvascular Hyperpermeability and Cardiovascular Arrhythmias

Background: Binge drinking combined with trauma is a major cause of mortality in US. It is known that acute alcohol intoxication (AAI) causes deleterious electrical changes in the heart, including involving QTc dispersion. We also recently demonstrated that acute alcohol intoxication (AAI) increases microvascular permeability. In the current study we investigated the consequences of combined hemorrhagic shock and resuscitation (HSR) and AAI on both microvascular permeability and cardiac electrical activity. Hypothesis and Methods: We hypothesized that AAI combined with HSR causes microvascular leakage and electrical dysfunction in the heart. We used an established rat model of AAI/HSR to test the microvascular and cardiac effects. Results: We identified that combined AAI/HSR significantly (p&lt;0.0007) prolongs the QTc interval (ms) compared to control water fed, sham rats (190.8±7.1 vs. 138.4±4.4, p&lt;0.05). Ex vivo acquired left ventricular monophasic action potential (AP) data revealed increased AP duration in the AAI/HSR group at 20 (9.7±0.6 vs. 5.5±0.2; p&lt;0.0004 ), 50 (16±1.1 vs. 8.8±0.6; p&lt;0.001), and 70% (24.7±2.4 vs. 19.5±0.5; p&lt;0.03) repolarization level relative to water/sham controls. Microvascular hyperpermeability was apparent in the mesenteric microcirculation in the AAI/HSR group compared to AAI/sham, water/HSR, and water/sham controls (IOI of 1061.7±208 HSR-alcohol vs. 698.4±311 HSR-water, p&lt;0.05; vs. 209.5±42 sham-alcohol, p&lt;0.001; and vs. 62.3±9 sham-water, p&lt;0.001). Conclusions: We report for the first time the cardiovascular hallmarks of AAI/HSR associated injury, which include prolonged QTc and AP durations and elevated microvascular permeability.

Abstract 14577: Kvβ1.1 Binds Pyridine Nucleotides and Alters Cardiac Action Potential

Introduction: Kvβ1 subunits demonstrate a unique ability to bind pyridine nucleotides and alter Kv channel function. Pyridine nucleotide levels are altered in cardiovascular diseases resulting in higher NADH levels. Therefore coupling of the Kvβ1 subunit with Kv channels may be important in how the repolarization is modulated in presence of NADH. Hypothesis: The Kvβ1 subunits are highly expressed in the heart and bind to multiple Kv channels (Kv4.x and Kv1.x) and regulate Kv channel activity. Therefore, we evaluated pyridine nucleotide changes in cardiovascular disease in the presence and absence of Kvβ1 and how they modulate Kv channel activity. Methods: Wild type (WT) and Kvβ1 knockout (KO) mice hearts (n=7 each group) using the ex-vivo Langendorff system were exposed to both acute and chronic alterations of lactate to induce an increase in NADH levels. Cardiac monophasic action potentials (MAPs) were measured before and 10 min after perfusion of the lactate, for chronic alterations mice were injected with isoprenaline to induce cardiac hypertrophy and demonstrated significant NADH increase. In addition we performed pull-down assays to identify binding partners of Kvβ1.1 in the heart. Results: We observed that, lactate exposure (10mmole/L) ( p &lt;0.05) prolonged left ventricular MAP duration in WT (9.4±0.7 vs. 15.6±1.2* at APD50) however KO mice demonstrated no significant difference (10.9±0.5 vs. 11.7±1.1). NADH levels in lactate exposed hearts demonstrated a 3 fold increase compared with control Krebs buffer. Lactate prolonged MAP durations in WT hearts, which was reversal by adding pyruvate (20mmole/L) to its control (pre-lactate; 8±0.5 vs. 13.6±0.6* vs. 8.4±0.6). The chronic model, MAP duration significantly ( p &lt;0.01) prolonged in the WT (9.7±0.7 vs. 15.3±1.1*) mouse hearts after isoprenaline infusion but not KO mouse (12.4±1.8 vs. 14.4±1). Immunoprecipitation experiments indicate that Kvβ1.1 interacts with Kv4.2 and Kv1.5 from mouse heart. Conclusions: Results obtained from this study suggested that the Kvβ1.1 subunit has the ability to sense redox changes including NAD/NADH alterations and modulate MAP through Kv activity. This study highlights the physiological role Kvβ1.1 plays in cardiac repolarization physiology and cardiac disease.

GW26-e2943 Effect of high extracellular calcium on left ventricular monophasic action potentials in Kunming mice

That the calcium ions across the membrane is an important part of the formation of action potential in myocardial cells, and the purpose of this experiment is to observe the effect of high calcium on the left ventricular monophasic action potential(MAP) in open chest model of Kunming mice. 10

The effect of stimulation frequency on the transmural ventricular monophasic action potential in yellowfin tuna Thunnus albacares

Monophasic action potentials (MAPs) were recorded from the spongy and compact layers of the yellowfin tuna Thunnus albacares ventricle as stimulation frequency was increased. MAP duration decreased with increase in stimulation frequency in both the spongy and compact myocardial layers, but no significant difference in MAP duration was observed between the layers.

High‐Rate Pacing Reduces Variability of Repolarization and Prevents Repolarization‐Dependent Arrhythmias in Dogs With Chronic AV Block

high-rate pacing may have an inhibitory effect on the initiation of Torsade de Pointes arrhythmias (TdP). However, permanent pacing is only indicated in high-risk patients. We performed a proof of concept study into automatic overdrive pacing for prevention of drug-induced TdP, using short-term variability of repolarization (STV) as a feedback parameter of arrhythmic risk. the minimal signal sampling frequency required for measuring STV was determined through computer simulation. Arrhythmogenic response to dofetilide (25 microg/kg/5 minutes) was tested at two different paced heart rates (60-65 bpm vs 100-110 bpm) in 7 dogs with chronic atrioventricular block, while recording right and left ventricular (LV) monophasic action potential (MAP) and LV electrogram (EGM). Simulations showed a sampling frequency of 500 Hz is sufficient to capture relevant STV values. High-rate pacing prevented dofetilide-induced TdP seen at the low rate (low: 6/7 vs high: 1/7). At the low rate, STV from LV MAP duration increased before occurrence of spontaneous, ectopic activity and TdP (1.7 ± 0.6-3.0 ± 1.8 ms, P < 0.05), but at the high-rate STV did not change significantly (0.9 ± 0.2-1.5 ± 1.4 ms, NS). Regression analysis showed a close relation between STV calculated from LV MAP and from LV EGM (R(2) = 0.71). high-rate pacing increases repolarization reserve in dogs with chronic atrioventricular block, preventing dofetilide-induced TdP. Changes in repolarization reserve are reflected in values of STV.

Robust anti‐arrhythmic efficacy of verapamil and flunarizine against dofetilide‐induced TdP arrhythmias is based upon a shared and a different mode of action

The high predisposition to Torsade de Pointes (TdP) in dogs with chronic AV-block (CAVB) is well documented. The anti-arrhythmic efficacy and mode of action of Ca(2+) channel antagonists, flunarizine and verapamil against TdP were investigated. Mongrel dogs with CAVB were selected based on the inducibility of TdP with dofetilide. The effects of flunarizine and verapamil were assessed after TdP and in different experiments to prevent dofetilide-induced TdP. Electrocardiogram and ventricular monophasic action potentials were recorded. Electrophysiological parameters and short-term variability of repolarization (STV) were determined. In vitro, flunarizine and verapamil were added to determine their effect on (i) dofetilide-induced early after depolarizations (EADs) in canine ventricular myocytes (VM); (ii) diastolic Ca(2+) sparks in RyR2(R4496+/+) mouse myocytes; and (iii) peak and late I(Na) in SCN5A-HEK 293 cells. Dofetilide increased STV prior to TdP and in VM prior to EADs. Both flunarizine and verapamil completely suppressed TdP and reversed STV to baseline values. Complete prevention of TdP was achieved with both drugs, accompanied by the prevention of an increase in STV. Suppression of EADs was confirmed after flunarizine. Only flunarizine blocked late I(Na). Ca(2+) sparks were reduced with verapamil. Robust anti-arrhythmic efficacy was seen with both Ca(2+) channel antagonists. Their divergent electrophysiological actions may be related to different additional effects of the two drugs.

Dose-Related Shortening of Ventricular Tachycardia Cycle Length After Administration of the KATP Channel Opener Bimakalim in a 4-Day-Old Chronic Infarct Anesthetized Pig Model

Potassium channel openers are known to act on potassium ATP-dependent channels in cardiac tissue. Such agents may exacerbate acceleration of acute ischemia-induced ventricular repolarization and aggravate arrhythmias. To test whether activation of K( ATP) channels during the healing period of myocardial infarction (MI) can still influence the electrophysiologic properties and the type of inducible arrhythmias, we investigated the effects of bimakalim (BIM) on sustained ventricular tachycardia (VT) 4 days after ligation of the left anterior descending (LAD) coronary artery in pigs. Programmed stimulation was performed to elicit VT prior to and after intravenous (IV) BIM. Combination monophasic action potential (MAP)/PACING catheters were used to enable simultaneous ventricular MAP recording and pacing. Ventricular effective refractory period (ERP) and MAP duration determined at 50% and 90% repolarization were measured prior to and after BIM. After completion of baseline measurements, BIM was consecutively given at 0.5, 1, and 3 mg/kg bolus followed by 0.025, 0.05, and 0.1 mg/kg per minute maintenance infusion, respectively. From a total of 23 pigs subjected to LAD ligation, 4 animals succumbed to infarction and the remaining 19 animals were studied by programmed stimulation. Only animals that exhibited reproducible and hemodynamically stable monomorphic VTs during control stimulation were selected for evaluation (n = 14). After the first, second, and third dose of BIM, the mean VT rate was increased by 6%, 14% (P <. 01), and 47% (P < .001) compared to control values, respectively. Ventricular ERP and repolarization were significantly shortened only by the second and third dose of BIM. Of 14 pigs receiving the highest BIM dosage, 3 revealed polymorphic VTs degenerating into ventricular fibrillation (VF). Our data suggest that high BIM doses may lead to faster and more aggressive pacing-induced reentrant VTs after subacute MI. This is consistent with the drug-induced acceleration of ventricular repolarization with shortening of MAP duration and refractoriness.