Ilija Uzelac

The Potential of Dual Camera Systems for Multimodal Imaging of Cardiac Electrophysiology and Metabolism

Fluorescence imaging has become a common modality in cardiac electrodynamics. A single fluorescent parameter is typically measured. Given the growing emphasis on simultaneous imaging of more than one cardiac variable, we present an analysis of the potential of dual camera imaging, using as an example our straightforward dual camera system that allows simultaneous measurement of two dynamic quantities from the same region of the heart. The advantages of our system over others include an optional software camera calibration routine that eliminates the need for precise camera alignment. The system allows for rapid setup, dichroic image separation, dual-rate imaging, and high spatial resolution, and it is generally applicable to any two-camera measurement. This type of imaging system offers the potential for recording simultaneously not only transmembrane potential and intracellular calcium, two frequently measured quantities, but also other signals more directly related to myocardial metabolism, such as [K+]e, NADH, and reactive oxygen species, leading to the possibility of correlative multimodal cardiac imaging. We provide a compilation of dye and camera information critical to the design of dual camera systems and experiments.

Regional increase of extracellular potassium leads to electrical instability and reentry occurrence through the spatial heterogeneity of APD restitution

The heterogeneities of electrophysiological properties of cardiac tissue are the main factors that control both arrhythmia induction and maintenance. Although the local increase of extracellular potassium ([K(+)](o)) due to coronary occlusion is a well-established metabolic response to acute ischemia, the role of local [K(+)](o) heterogeneity in phase 1a arrhythmias has yet to be determined. In this work, we created local [K(+)](o) heterogeneity and investigated its role in fast pacing response and arrhythmia induction. The left marginal vein of a Langendorff-perfused rabbit heart was cannulated and perfused separately with solutions containing 4, 6, 8, 10, and 12 mM of K(+). The fluorescence dye was utilized to map the voltage distribution. We tested stimulation rates, starting from 400 ms down to 120 ms, with steps of 5-50 ms. We found that local [K(+)](o) heterogeneity causes action potential (AP) alternans, 2:1 conduction block, and wave breaks. The effect of [K(+)](o) heterogeneity on electrical stability and vulnerability to arrhythmia induction was largest during regional perfusion with 10 mM of K(+). We detected three concurrent dynamics: normally propagating activation when excitation waves spread over tissue perfused with normal K(+), alternating 2:2 rhythm near the border of [K(+)](o) heterogeneity, and 2:1 aperiodicity when propagation was within the high [K(+)](o) area. [K(+)](o) elevation changed the AP duration (APD) restitution and shifted the restitution curve toward longer diastolic intervals and shorter APD. We conclude that spatial heterogeneity of the APD restitution, created with regional elevation of [K(+)](o), can lead to AP instability, 2:1 block, and reentry induction.

Mechanism for Amplitude Alternans in Electrocardiograms and the Initiation of Spatiotemporal Chaos.

It is widely believed that one major life-threatening transition to chaotic fibrillation occurs via spiral-wave breakup that is preceded by spatiotemporal dispersion of refractoriness due to alternations in the duration of the cardiac action potential (AP). However, recent clinical and experimental evidence suggests that other characteristics of the AP may contribute to, and perhaps drive, this dangerous dynamical instability. To identify the relative roles of AP characteristics, we performed experiments in rabbit hearts under conditions to minimize AP duration dynamics which unmasked pronounced AP amplitude alternans just before the onset of fibrillation. We used a simplified ionic cell model to derive a return map and a stability condition that elucidates a novel underlying mechanism for AP alternans and spiral breakup. We found that inactivation of the sodium current is key to developing amplitude alternans and is directly connected to conduction block and initiation of arrhythmias. Simulations in 2D where AP amplitude alternation led to turbulence confirm our hypothesis.

Electromechanical vortex filaments during cardiac fibrillation

The self-organized dynamics of vortex-like rotating waves, which are also known as scroll waves, are the basis of the formation of complex spatiotemporal patterns in many excitable chemical and biological systems. In the heart, filament-like phase singularities that are associated with three-dimensional scroll waves are considered to be the organizing centres of life-threatening cardiac arrhythmias. The mechanisms that underlie the onset, maintenance and control of electromechanical turbulence in the heart are inherently three-dimensional phenomena. However, it has not previously been possible to visualize the three-dimensional spatiotemporal dynamics of scroll waves inside cardiac tissues. Here we show that three-dimensional mechanical scroll waves and filament-like phase singularities can be observed deep inside the contracting heart wall using high-resolution four-dimensional ultrasound-based strain imaging. We found that mechanical phase singularities co-exist with electrical phase singularities during cardiac fibrillation. We investigated the dynamics of electrical and mechanical phase singularities by simultaneously measuring the membrane potential, intracellular calcium concentration and mechanical contractions of the heart. We show that cardiac fibrillation can be characterized using the three-dimensional spatiotemporal dynamics of mechanical phase singularities, which arise inside the fibrillating contracting ventricular wall. We demonstrate that electrical and mechanical phase singularities show complex interactions and we characterize their dynamics in terms of trajectories, topological charge and lifetime. We anticipate that our findings will provide novel perspectives for non-invasive diagnostic imaging and therapeutic applications.

Simultaneous Quantification of Spatially Discordant Alternans in Voltage and Intracellular Calcium in Langendorff-Perfused Rabbit Hearts and Inconsistencies with Models of Cardiac Action Potentials and Ca Transients

Rationale: Discordant alternans, a phenomenon in which the action potential duration (APDs) and/or intracellular calcium transient durations (CaDs) in different spatial regions of cardiac tissue are out of phase, present a dynamical instability for complex spatial dispersion that can be associated with long-QT syndrome (LQTS) and the initiation of reentrant arrhythmias. Because the use of numerical simulations to investigate arrhythmic effects, such as acquired LQTS by drugs is beginning to be studied by the FDA, it is crucial to validate mathematical models that may be used during this process. Objective: In this study, we characterized with high spatio-temporal resolution the development of discordant alternans patterns in transmembrane voltage (Vm) and intracellular calcium concentration ([Cai]+2) as a function of pacing period in rabbit hearts. Then we compared the dynamics to that of the latest state-of-the-art model for ventricular action potentials and calcium transients to better understand the underlying mechanisms of discordant alternans and compared the experimental data to the mathematical models representing Vm and [Cai]+2 dynamics. Methods and Results: We performed simultaneous dual optical mapping imaging of Vm and [Cai]+2 in Langendorff-perfused rabbit hearts with higher spatial resolutions compared with previous studies. The rabbit hearts developed discordant alternans through decreased pacing period protocols and we quantified the presence of multiple nodal points along the direction of wave propagation, both in APD and CaD, and compared these findings with results from theoretical models. In experiments, the nodal lines of CaD alternans have a steeper slope than those of APD alternans, but not as steep as predicted by numerical simulations in rabbit models. We further quantified several additional discrepancies between models and experiments. Conclusions: Alternans in CaD have nodal lines that are about an order of magnitude steeper compared to those of APD alternans. Current action potential models lack the necessary coupling between voltage and calcium compared to experiments and fail to reproduce some key dynamics such as, voltage amplitude alternans, smooth development of calcium alternans in time, conduction velocity and the steepness of the nodal lines of APD and CaD.

Cardiac arrhythmia recognition with robust discrete wavelet-based and geometrical feature extraction via classifiers of SVM and MLP-BP and PNN neural networks

Introduction: An ECG signal has important information that can help for reflecting cardiac activity of a patient and medical diagnosis. Consistent or periodical heart rhythm disorders can result cardiac arrhythmias so classification algorithm for recognizing arrhythmias with satisfactory accuracy is necessary. Aims: In this study, a robust wavelet based algorithm for detection and delineation of events in ECG signal is applied and then a new synthesis of MLP-BP and PNN neural networks for heart arrhythmia classification was described Methods: As a matter of fact any changes in the morphology of an ECG due to the arrhythmia are observed in time and frequency analysis so multi resolution analysis is applied for feature detection. First, noise and artifact is rejected by a discrete wavelet transform (DWT) and multi lead ECG is obtained Then QRS complexes of signal is extracted and the signal is decomposed so corresponding DWT scales are segmented Next curve length and high order moment order based feature extraction are calculated for each excerpted segment and elements of feature vector for regulating the parameters of classifiers are obtained After generation of feature source and segmentation, Multi-Layer Perceptron-Back Propagation (MLP-BP) neural networks, Probabilistic Neural Network (PNN) and support vector machine (SVM) were designed and tuned and their results were compared Results: The proposed algorithm was tested to all 48 record of the MIT-BIH arrhythmia database and also the proposed topology of classifiers and its related parameters is optimized by searching of best value of parameters. The average value of accuracy of each classifier over all records of MIT-BIH for arrhythmias recognition is Acc=97.42, Acc=98. 24 and Acc=97. 42 for S VM, MLP and PNN classifiers respectively and also obtained results were compared with similar peer-reviewed studies in this subject.

A novel method for arterial blood pressure pulse detection based on a new coupling strategy and discrete wavelet transform

In this study, a new method for detection of arterial blood pressure pulses (ABP) is presented. The algorithm employs discrete wavelet transform (DWT) decomposition to extract ABP waveform features. In the proposed method, two strategies are used. In the first strategy, the algorithm uses only the DWT coefficients of ABP. The second strategy which is introduced for the first time in this paper, the coupled DWT coefficients of ABP and electrocardiogram (ECG) are used. When DWT coefficients of ECG and ABP are coupled the detection of ABP pulses is easier and more accurate in noisy parts of the signals. Furthermore, this coupling strategy is useful for the detection of ECG R-peaks since the ABP pulses make the ECG R-peaks detectable when ECG is noisy. To meet this end, adaptive thresholding and different DWT functions were employed and fitted because of different morphologies of ABP and ECG signals. The ABP-ECG delay time is measured in different recordings and set for 254 milliseconds. For evaluation, 170 recordings of the multimodal training set of PhysioNet/Computing in Cardiology Challenge 2014 which contained both ECG and ABP waveforms were used. The first and second strategy obtained average accuracy of 87.56% and 88.53%, respectively.

Robust framework for quantitative analysis of optical mapping signals without filtering

Experimental studies with in-vitro isolated hearts using optical mapping techniques have had a significant impact on our understanding of cardiac electrophysiology. The trans-membrane voltage Vm, and intracellular free calcium concentration [Cai]+2 signals obtained from optical mapping experiments can often be corrupted with noise. This is mostly due to the small light intensities and very short exposure times when high speed cameras are used at frames rates of 500-1000 fps. In addition, for small preparations or recordings of small areas, the noise floor levels are even greater and can be comparable to the amplitude of the signal (S/N ≈1). In general strong spatial and temporal filtering is necessary to remove the noise at the expenses of signal degradation and loss of critical information especially at high frequencies that are of a particular interest. In this paper we present and analyze an oversampling image processing technique where due to the cycling processes in cardiac activity during steady state dynamics we are able to stack (sum up) the images recorded at specific equidistant time intervals. The stacking process reduces the noise effectively as the square root of the number of stacked images used. We show that no spatial or temporal filtering is needed to obtain useful data with the stacking technique that allows us to resolve information on a time scale only limited with a sampling rate.

Diastolic Field Stimulation: the Role of Shock Duration in Epicardial Activation and Propagation

Detailed knowledge of tissue response to both systolic and diastolic shock is critical for understanding defibrillation. Diastolic field stimulation has been much less studied than systolic stimulation, particularly regarding transient virtual anodes. Here we investigated high-voltage-induced polarization and activation patterns in response to strong diastolic shocks of various durations and of both polarities, and tested the hypothesis that the activation versus shock duration curve contains a local minimum for moderate shock durations, and it grows for short and long durations. We found that 0.1-0.2-ms shocks produced slow and heterogeneous activation. During 0.8-1 ms shocks, the activation was very fast and homogeneous. Further shock extension to 8 ms delayed activation from 1.55 ± 0.27 ms and 1.63 ± 0.21 ms at 0.8 ms shock to 2.32 ± 0.41 ms and 2.37 ± 0.3 ms (N = 7) for normal and opposite polarities, respectively. The traces from hyperpolarized regions during 3-8 ms shocks exhibited four different phases: beginning negative polarization, fast depolarization, slow depolarization, and after-shock increase in upstroke velocity. Thus, the shocks of >3 ms in duration created strong hyperpolarization associated with significant delay (P < 0.05) in activation compared with moderate shocks of 0.8 and 1 ms. This effect appears as a dip in the activation-versus-shock-duration curve.

Electrocardiogram reconstruction from high resolution voltage optical mapping

Electrocardiogram recordings during opucal mapping experiments in heart tissue are commonly used tu monitor the health of the preparation and to obtain dominant frequencies during arrhythmic and defibrillatory studies. However the use of ECG reconstructed from optical mapping is seldom used and to date it has not been strictly validated. In this manuscript we present the first detailed validation and comparison of Optical Mapping ECG, or OM-ECG, with standard ECG recordings by calculating the electrostatic potential in space as a function of the voltage measured optically and describe the different approximations that can be used to obtain unipolar or bipolar ECG recordings. We found that in small/medium hearts, such as rabbits, leads that are aligned apex to base only require activation recording from one surface (anterior or posterior) for the OM-ECG to match the ECG while leads aligned left to right may require both an anterior and posterior optical mapping recording. The discrepancy between leads is due to symmetries in the ventricular activations. In the case of ischemic hearts where activations even-out more, the match between the OM-ECG and standard ECG may require only one surface recording for both left-right and base-apex leads. We believe that this methodology has two main and direct applications in the study of cardiac dynamics. The first is during studies of defibrillation where information after the shock may be crucial in the development of new strategies, OM-ECGs do not suffer the current artifacts of standard ECGs during shocks and can be calculated during the entire activation. We present examples in rabbit ventricles where even low amplitude pacing artifacts are captured by the ECG but do not appear in the OM-ECG. The second use of this technique is for reconstructions of intramural dynamics in larger hearts where differences between the ECG and OM-ECG obtained from anterior and posterior recordings can be used to derive the intramural activation.Electrocardiogram recordings during opucal mapping experiments in heart tissue are commonly used tu monitor the health of the preparation and to obtain dominant frequencies during arrhythmic and defibrillatory studies. However the use of ECG reconstructed from optical mapping is seldom used and to date it has not been strictly validated. In this manuscript we present the first detailed validation and comparison of Optical Mapping ECG, or OM-ECG, with standard ECG recordings by calculating the electrostatic potential in space as a function of the voltage measured optically and describe the different approximations that can be used to obtain unipolar or bipolar ECG recordings. We found that in small/medium hearts, such as rabbits, leads that are aligned apex to base only require activation recording from one surface (anterior or posterior) for the OM-ECG to match the ECG while leads aligned left to right may require both an anterior and posterior optical mapping recording. The discrepancy between leads is due to symmetries in the ventricular activations. In the case of ischemic hearts where activations even-out more, the match between the OM-ECG and standard ECG may require only one surface recording for both left-right and base-apex leads. We believe that this methodology has two main and direct applications in the study of cardiac dynamics. The first is during studies of defibrillation where information after the shock may be crucial in the development of new strategies, OM-ECGs do not suffer the current artifacts of standard ECGs during shocks and can be calculated during the entire activation. We present examples in rabbit ventricles where even low amplitude pacing artifacts are captured by the ECG but do not appear in the OM-ECG. The second use of this technique is for reconstructions of intramural dynamics in larger hearts where differences between the ECG and OM-ECG obtained from anterior and posterior recordings can be used to derive the intramural activation.