Shahriar Iravanian

Cardiomyocyte Cultures With Controlled Macroscopic Anisotropy. A Model for Functional Electrophysiological Studies of Cardiac Muscle

Abstract— Structural and functional cardiac anisotropy varies with the development, location, and pathophysiology in the heart. The goal of this study was to design a cell culture model system in which the degree, change in fiber direction, and discontinuity of anisotropy can be controlled over centimeter-size length scales. Neonatal rat ventricular myocytes were cultured on fibronectin on 20-mm diameter circular cover slips. Structure-function relationships were assessed using immunostaining and optical mapping. Cell culture on microabraded cover slips yielded cell elongation and coalignment in the direction of abrasion, and uniform, macroscopically continuous, elliptical propagation with point stimulation. Coarser microabrasion (wider and deeper abrasion grooves) increased longitudinal (23.5 to 37.2 cm/s;r =0.66) and decreased transverse conduction velocity (18.1 to 9.2 cm/s;r =−0.84), which resulted in increased longitudinal-to-transverse velocity anisotropy ratios (1.3 to 3.7, n=61). A thin transition zone between adjacent uniformly anisotropic areas with 45° or 90° difference in fiber orientation acted as a secondary source during 2× threshold field stimulus. Cell culture on cover slips micropatterned with 12- or 25-&mgr;m wide fibronectin lines and previously coated with decreasing concentrations of background fibronectin yielded transition from continuous to discontinuous anisotropic architecture with longitudinally oriented intercellular clefts, decreased transverse velocity (16.9 to 2.6 cm/s;r =−0.95), increased velocity anisotropy ratios (1.6 to 5.6, n=70), and decreased longitudinal velocity (36.4 to 14.6 cm/s;r =−0.85) for anisotropy ratios >3.5. Cultures of cardiac myocytes with controlled degree, uniformity and continuity of structural, and functional anisotropy may enable systematic 2-dimensional in vitro studies of macroscopic structure-related mechanisms of reentrant arrhythmias. The full text of this article is available at http://www.circresaha.org.

The renin-angiotensin-aldosterone system (RAAS) and cardiac arrhythmias.

The role of the renin-angiotensin-aldosterone system (RAAS) in many cardiovascular disorders, including hypertension, cardiac hypertrophy, and atherosclerosis, is well established, whereas its relationship with cardiac arrhythmias is a new area of investigation. Atrial fibrillation and malignant ventricular tachyarrhythmias, especially in the setting of cardiac hypertrophy or failure, seem to be examples of RAAS-related arrhythmias because treatment with RAAS modulators, including angiotensin-converting enzyme inhibitors, angiotensin receptor blockers, and mineralocorticoid receptor blockers, reduces the incidence of these arrhythmias. RAAS has a multitude of electrophysiological effects and can potentially cause arrhythmia through a variety of mechanisms. We review new experimental results that suggest that RAAS has proarrhythmic effects on membrane and sarcoplasmic reticulum ion channels and that increased oxidative stress is likely contributing to the increased arrhythmic incidence. A summary of ongoing clinical trials that will address the clinical usefulness of RAAS modulators for prevention or treatment of arrhythmias is presented.

Inhibition of c-Src tyrosine kinase prevents angiotensin II-mediated connexin-43 remodeling and sudden cardiac death.

OBJECTIVES The aim of this study was to test whether c-Src tyrosine kinase mediates connexin-43 (Cx43) reduction and sudden cardiac death in a transgenic mouse model of cardiac-restricted overexpression of angiotensin-converting enzyme (ACE8/8 mice). BACKGROUND Renin-angiotensin system activation is associated with an increased risk for arrhythmia and sudden cardiac death, but the mechanism is not well understood. The up-regulation of c-Src by angiotensin II may result in the reduction of Cx43, which impairs gap junction function and provides a substrate for arrhythmia. METHODS Wild-type and ACE8/8 mice with and without treatment with the c-Src inhibitor 1-(1,1-dimethylethyl)-1-(4-methylphenyl)-1H-pyrazolo[3,4-d]pyrimidin-4-amine (PP1) were studied. Telemetry monitoring, in vivo electrophysiologic studies, Western blot analyses for total and phosphorylated c-Src and Cx43, immunohistochemistry staining for Cx43, and functional assessment of Cx43 with fluorescent dye diffusion were performed. RESULTS The majority of the arrhythmic deaths resulted from ventricular tachycardia degenerating to ventricular fibrillation (83%). Levels of total and phosphorylated c-Src were increased and Cx43 reduced in ACE8/8 mice. PP1 reduced total and phosphorylated c-Src levels, increased Cx43 level by 2.1-fold (p < 0.005), increased Cx43 at the gap junctions (immunostaining), improved gap junctional communication (dye spread), and reduced ventricular tachycardia inducibility and sudden cardiac death. The survival rate increased from 11% to 86% with 4 weeks of PP1 treatment (p < 0.005). Treatment with an inactive analog did not change survival or Cx43 levels. CONCLUSIONS Renin-angiotensin system activation is associated with c-Src up-regulation, Cx43 loss, reduced myocyte coupling, and arrhythmic sudden death, which can be prevented by c-Src inhibition. This suggests that an increase in c-Src activity may help mediate renin-angiotensin system-induced arrhythmias and that c-Src inhibitors might exert antiarrhythmic activity.

A novel algorithm for cardiac biosignal filtering based on filtered residue method

In this paper, a new algorithm is presented for the filtering (de-noising) of cardiac bioelectrical signals. The primary target of this algorithm is the class of cardiac action potentials recorded using voltage-sensitive dyes, although the method is also applied to electrocardiographic signals High periodicity is one of the main features of cardiac biosignals. The proposed algorithm exploits this feature in filtering signals with a minimum amount of distortion. The basic idea is to use signal averaging in time to rind the stationary portion of the signal. The residue is found by subtracting the signal average from the corresponding points of the input. After passing through a low-pass filter, the filtered residue (FR) is added back to the signal average to reconstruct the output. The practical implementation of the filter residue algorithm is discussed. Stretching and shrinking operations are the basis for the conversion of quasi-periodic signals into periodic signals, which can then be subjected to the FR algorithm. Various examples are presented, and error estimation is performed to guide the selection of optimal parameters for the algorithm. The ability of the algorithm to reconstruct the variation among beats is demonstrated, and its limitations are discussed.

Control of Action Potential Duration Alternans in Canine Cardiac Ventricular Tissue

Cardiac electrical alternans, characterized by a beat-to-beat alternation in action potential waveform, is a naturally occurring phenomenon, which can occur at sufficiently fast pacing rates. Its presence has been putatively linked to the onset of cardiac reentry, which is a precursor to ventricular fibrillation. Previous studies have shown that closed-loop alternans control techniques that apply a succession of externally administered cycle perturbations at a single site provide limited spatially-extended alternans elimination in sufficiently large cardiac substrates. However, detailed experimental investigations into the spatial dynamics of alternans control have been restricted to Purkinje fiber studies. A complete understanding of alternans control in the more clinically relevant ventricular tissue is needed. In this paper, we study the spatial dynamics of alternans and alternans control in arterially perfused canine right ventricular preparations using an optical mapping system capable of high-resolution fluorescence imaging. Specifically, we quantify the spatial efficacy of alternans control along 2.5 cm of tissue, focusing on differences in spatial control between different subregions of tissue. We demonstrate effective control of spatially-extended alternans up to 2.0 cm, with control efficacy attenuating as a function of distance. Our results provide a basis for future investigations into electrode-based control interventions of alternans in cardiac tissue.

Oxidative Stress and the Pathogenesis of Atrial Fibrillation

There is growing evidence that oxidative stress is involved in the pathogenesis of atrial fibrillation. Many known triggers of oxidative stress, such as age, diabetes, smoking, inflammation, and renin angiotensin system activation are linked with an increased risk of the arrhythmia. Blockers of angiotensin II signaling and other drugs with anti-oxidant properties can reduce the incidence of atrial fibrillation. Now, studies in animal models and human tissue have shown di- rectly that atrial fibrillation is associated with increased atrial oxidative stress. We review the evidence for a role of oxida- tive stress in causing atrial fibrillation and propose a unifying hypothesis that multiple triggers elicit oxidative stress which acts to enhance the risk of atrial fibrillation through ion channel dysregulation.

Relationship between mechanical dyssynchrony and intra-operative electrical delay times in patients undergoing cardiac resynchronization therapy

BackgroundIt is important to understand the relationship between electrical and mechanical ventricular activation in CRT patients. By measuring local electrical activation at multiple locations within the coronary veins and myocardial contraction at the same locations in the left ventricle, we determined the relationship between electrical and mechanical activation at potential left ventricular pacing locations.MethodsIn this study, mechanical contraction times were computed using high temporal resolution cine cardiovascular magnetic resonance (CMR) data, while electrical activation times were derived from intra-procedural local electrograms.ResultsIn our cohort, there was a strong correlation between electrical and mechanical delay times within each patient (R2 = 0.78 ± 0.23). Additionally, the latest electrically activated location corresponded with the latest mechanically contracting location in 91% of patients.ConclusionsThis study provides initial evidence that our method of obtaining non-invasive mechanical activation patterns accurately reflects the underlying electromechanical substrate of intraventricular dyssynchrony.

Representation of Collective Electrical Behavior of Cardiac Cell Sheets

The electrocardiogram (ECG) is a measure of the collective electrical behavior of the heart based on body surface measurements. With computational models or tissue preparations, various methods have been used to compute the pseudo-ECG (pECG) of bipolar and unipolar leads that can be given clinical interpretation. When spatial maps of transmembrane potential (V(m)) are available, pECG can be derived from a weighted sum of the spatial gradients of V(m). The concept of a lead field can be used to define sensitivity curves for different bipolar and unipolar leads and to determine an effective operating height for the bipolar lead position for a two-dimensional sheet of heart cells. The pseudo-vectorcardiogram (pVCG) is computed from orthogonal bipolar lead voltages, which are derived in this study from optical voltage maps of cultured monolayers of cardiac cells. Rate and propagation direction for paced activity, rotation frequency for reentrant activity, direction of the common pathway for figure-eight reentry, and transitions from paced activity to reentry can all be distinguished using the pVCG. In contrast, the unipolar pECG does not clearly distinguish among many of the different types of electrical activity. We also show that pECG can be rapidly computed by two geometrically weighted sums of V(m), one that is summed over the area of the cell sheet and the other over the perimeter of the cell sheet. Our results are compared with those of an ad hoc difference method used in the past that consists of a simple difference of the sum of transmembrane potentials on one side of a tissue sheet and that of the other.

Control of action potential duration alternans in canine ventricular tissue

Cardiac action potential duration alternans is characterized by a beat-to-beat alternation in action potential waveform. Its presence has been putatively linked to the onset of lethal cardiac arrhythmias. Previous studies, which have been limited to cardiac Purkinje fibers, have shown that closed-loop alternans control techniques, which apply a succession of externally administered cycle perturbations, provide ineffectual spatial alternans elimination. A more complete understanding of alternans control in the more clinically relevant ventricular tissue is needed. Here, we study the spatial dynamics of alternans and alternans control in arterially perfused canine right ventricular preparations using optical mapping. We quantified the spatial efficacy of alternans control across 2.5 cm of tissue, focusing primarily on differences in spatial control within several sub-regions of tissue. Our results provide a basis for future investigations into multi-electrode-based control interventions of alternans in cardiac tissue.

Spatiotemporal organization during ablation of persistent atrial fibrillation

BACKGROUND Targeting complex fractionated atrial electrograms improves the outcome of ablation of persistent atrial fibrillation (AF); however, the mechanism(s) responsible for the generation of complex fractionated atrial electrogram signals and efficacy of ablation is not clear. OBJECTIVE The aim of this study was to gain mechanistic insight into ablation of persistent AF by evaluating the spatiotemporal patterns of atrial organization during ablation. METHODS Intracardiac recordings from 18 ablation procedures were analyzed. Signals recorded by right atrial/coronary sinus catheters were processed. We quantified atrial organization using recurrence maps and recurrence percentage (Rec%) methodology and generated temporally dense time series of cycle lengths and Rec%. RESULTS A total of 162 intra-atrial recordings were categorized into type I (sudden jump in Rec%), type II (gradual increase), and type III (no increase). Type I was the most common form and was seen in 57% ± 4% of the recordings. A typical pattern was the initial appearance of local organization, which then expanded to adjacent channels in discrete jumps until eventually an organized atrial flutter emerged. This pattern is consistent with the atrial organization signature expected from ablation of a single spiral wave with fibrillatory conduction to the rest of atria. CONCLUSION Temporally dense spatiotemporal assessment of atrial organization during the ablation of persistent AF is feasible and provides complementary information to cycle length measurements. Atrial organization starts locally and expands spatially in discrete jumps. The regularization of AF to atrial flutter exhibits characteristics of phase transition in complex systems.