Closed-chest Pigs Research Articles

Open- vs. closed-chest pig models of donation after circulatory death

Open- vs. closed-chest pig models of donation after circulatory death

Real-time electrophysiological characterization of acute and chronic radiofrequency ablation lesions

Abstract Background Assessment of lesion formation remains one of the most important goals to guide radiofrequency (RF) procedures in the cardiac electrophysiology laboratory. Cardiac navigation systems use specific algorithms to predict the effective lesion size, but these metrics are based only on ablation parameters and ignore local electric tissue characteristics. [1] Recent studies show that local multiparametric impedance is influenced by the intrinsic structural characteristics of the tissue. [2] Purpose This study aimed to assess the ability of local multiparametric impedance to characterize acute and chronic RF ablation lesions in the right atrium of pigs. Methods Four anesthetized closed-chest pigs were submitted to two interventions. The first intervention aimed at creating RF ablation lesions (30W, 60s, 55°) in the right atrium using an electrocatheter connected to a conventional cardiac navigation system, while mapping the local multiparametric impedance (LMI), the generator impedance (GI) and the bipolar voltage (BiV). All RF ablation lesions were performed with a stable contact force. Four weeks later, the second intervention aimed at mapping again the previous ablation sites. After the second intervention animals were euthanized and the hearts were removed and processed to identify the presence of fibrosis in the previously ablated lesions (Figure). Results We performed 19 ablations (5±2 per animal) with an average contact force of 14.2±4.6g. Four weeks later, 14/19 (74%) ablation lesions were identified as fibrotic points in the explanted heart (Figure 1). These showed decreased acute LMI and BiV values that persisted low after 4 weeks (Table 1). The remaining 5/19 ablations that did not present fibrosis had lower baseline LMI and bipolar values that resulted in lower LMI and BiV drops (Table 1). The absolute drop in LMI between effective and non-effective lesions was around 50%, while GI drop was only 21% (LMI drop @51KHz: From −3.0±1.4° to −1.5±0.3°, T-TEST p<0.05; GI drop: From 12.7±27.6Ω to 10.5±21.3Ω, T-TEST p=0.89). Conclusion The local baseline electrical properties of atrial myocardium are directly related to the effectiveness of RF ablation lesions. RF ablation sites that resulted into persistent fibrosis points had larger LMI and BiV values, with higher absolute drops than non-effective lesions. The use of LMI in clinical practice could improve the outcome of the procedures of arrhythmia ablation in the electrophysiology lab. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Supported by grants from ISCI-MINECO (FIS PI21/00392), FEDER, CIBERCV (CB16/11/00276)

Graded lower body negative pressure induces intraventricular negative pressures and incremental diastolic suction: a pressure-volume study in a porcine model.

Lower body negative pressure (LBNP) is a tool to study compensatory mechanisms to central hypovolemia for decades. However, the underlying hemodynamic mechanisms were mostly assessed noninvasively and remain unclear. We hypothesized that incremental LBNP reduces diastolic filling and thereby affects left ventricular (LV) diastolic suction (DS). Here, we investigated the impact of graded LBNP at three different levels of seal as well as during β-adrenergic stimulation by invasive pressure-volume (PV) analysis. Eight Landrace pigs were instrumented closed-chest for PV assessment. LBNP was applied at three consecutive locations: I) cranial, 10 cm below xiphoid process; II) medial, half-way between cranial and caudal; III) caudal, at the iliac spine. Level III was repeated under dobutamine infusion. At each level, baseline measurements were followed by application of incremental LBNP of -15, -30, and -45 mmHg. LBNP induced varying degrees of preload-dependent hemodynamic changes, with cranial LBNP inducing more pronounced effects than caudal. According to the Frank-Starling mechanism, graded LBNP progressively reduced LV stroke volume (LV SV) following a decrease in LV end-diastolic volume. Negative intraventricular minimal pressures were observed during dobutamine-infusion as well as higher levels of LBNP. Of note, incremental LV negative pressures were accompanied by increasing DS volumes, derived by extrapolating the volume at zero transmural pressure, the so-called equilibrium volume (V0), related to LV SV. In conclusion, graded preload reduction via LBNP shifts the PV loop to smaller volumes and end-systolic volume below V0, which induces negative LV pressures and increases LV suction. Accordingly, LBNP-induced central hypovolemia is associated with increased DS.NEW & NOTEWORTHY This study examined the effects of incremental lower body negative pressure (LBNP) from -15 to -45 mmHg on hemodynamic regulation using invasive pressure-volume assessment in closed-chest pigs. Graded preload reduction via LBNP induces negative left ventricular (LV) pressures while increasing LV suction and thus allowing the ventricle to eject below the equilibrium volume at the end of systole. Accordingly, LBNP-induced central hypovolemia is associated with increased diastolic suction.

PO-710-06 ASSESSMENT OF LESION DURABILITY PRODUCED BY PULSED ELECTRIC FIELD ABLATION: EVALUATION IN A SWINE ATRIAL LINEAR LESION MODEL

Pulsed electric field (PEF) ablation may cause durable or transient injury to cardiac myocytes. To examine acute and chronic integrity of the line of conduction block produced by PEF ablation in a swine heart model. A 7F catheter with a 3.5 mm ablation electrode (TactiCath, Abbott) was connected to a PEF generator (CENTAURI, Galaxy Medical) and positioned in RA and LA under EnSite guidance in 4 closed chest pigs. Using the lowest energy output settings of the PEF system (19 A, 1.4 ms), biphasic PEF current was delivered between the ablation electrode and a skin patch. A total of 10 linear ablations were performed in the SVC (n=3), inter-caval region (n=4) and right superior pulmonary vein (n=3) using point-by-point ablation (average CF: 15 g) with different inter-lesion distances (ILD): 12-15 mm (n=1), 6-8 mm (n=5) and 4 mm (n=4). Conduction block/isolation was assessed acutely (4 pigs) and 14 days (3 pigs) after index procedures. Complete elimination of electrograms on the ablation catheter was typically observed after the first PEF application. Complete line of block/isolation was obtained acutely in all 10 lines, confirmed by EnSite activation and voltage maps. At 14 days, recovery of conduction was observed in 4/5 (80%) lines with 6-8 mm ILD and 0/4 lines with 4 mm ILD (p<0.05). Gaps in linear lesions were confirmed by gross pathology in 4 lines with reconnection using TTC staining. At the lowest PEF dose, acute conduction block can be consistently achieved with large ILD. While all 4 mm ILD lesions remained durable, recovery of conduction at 14 days was often observed for ILD≥ 6 mm, suggesting durable lesion sizes between 4-5 mm surrounded by transiently stunned myocardium.

PO-710-03 TEMPORAL AND SPATIAL EVOLUTION OF REVERSIBLE AND IRREVERSIBLE VENTRICULAR LESION BOUNDARIES PRODUCED BY PULSED ELECTRIC FIELD ABLATION

During pulsed electric field (PEF) ablation, electric pulses induce reversible (R) or irreversible (IR) injury on cardiac myocytes. Using triphenyl tetrazolium chloride (TTC) staining, we have reported that acute PEF ablation lesions show a dark central zone (DCZ, contraction band necrosis and hemorrhage) surrounded by pale boundary (PB, no mitochondrial activity, consistent with IR zone) and a hyperstained rim (HR, indicating enhanced mitochondrial activity) beyond the PB, indicating R zone. To assess histologically the spatial extent and temporal evolution of PEF ablation lesions in a swine model. A 7Fr catheter with a 3.5 mm ablation electrode (TactiCath, Abbott) was connected to a PEF generator (CENTAURI, Galaxy Medical) and positioned in RV and LV under EnSite guidance in 4 closed chest pigs. Biphasic PEF current (19 A, total duration 1.4 ms) was delivered between the ablation electrode and a skin patch at 50 ventricular sites at different target timepoints of <30 min, 1hr, 2hrs, 4hrs and 14 days after ablation. TTC was infused prior to animal sacrifice. Fig. The HR was initially greatest (<30 min) and gradually decreased over time while PB increased up to 4hr after ablation. The DCZ became evident only after 1hr and increased up to 4hr after ablation. At 14 days, ablation lesions show slightly smaller PB with no or minimal DCZ and HR. Histology showed fibrosis within the PB at 14 days, consistent with IR zone. TTC staining showed dynamic temporal and spatial evolution of the DCZ, PB and HR. The large initial HR zone indicates IR lesions are surrounded by relatively large R zone even 4hrs after ablation, suggesting the mechanism of transient conduction block during PEF ablation.

Externally-induced shear waves in the right ventricular free wall throughout the cardiac cycle

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): 1. NWO STW/Dutch Heart foundation 2. NWO Vidi TTW Background The right ventricular (RV) size and function have prognostic value, not only in right heart conditions (pulmonary hypertension, congenital heart disease), but also in left ventricular (LV) disease states. There is currently no noninvasive method for measuring the elastic properties of the RV walls. Increased RV diastolic stiffness however, as determined invasively, is linked to adverse prognosis in pulmonary arterial hypertension. In the LV, new high frame rate (HFR) echocardiography applications have been used in order to determine the propagation speed of naturally-occurring or externally-induced shear waves (SW) in the septum, which is related to myocardial stiffness. Purpose We show for the first time that SWs in the RV free wall (RVFW) can be externally induced and imaged transthoracically with ultrasound. Methods SW imaging was performed using a linear array with a programmable research ultrasound system, aimed at the parasternal long-axis view of a 5-weeks-old Yorkshire-Landrace pig (Fig. 1a). SWs were induced by a high intensity focused beam (f0 = 4.5 MHz) on the RVFW (Fig. 1a). This push beam generates a downwards force at the focus, which induces SWs that propagate left and right from the focus along the RVFW, visible on tissue Doppler images (TDI) shown in Figure 1b-c. SW propagation was imaged (f0 = 5.2 MHz) using HFR ultrasound (Fig. 1b, frame rate = 3 kHz). Three acquisitions of 1 second were performed, where 14 SWs were sequentially induced during each acquisition. ECG signal was captured simultaneously and synchronized offline. SW speeds were estimated using a custom, semi-automatic pipeline that includes TDI calculation, and SW speed estimation along a manually traced spline on the RVFW. This was repeated two times to include variability due to the manual processes. Up to four SW speed estimates were made after each push beam. SW speed estimation was repeated for all push pulses throughout the cardiac cycle, and the mean and standard deviation of the estimates were plotted (Fig. 2). Results At least 85% of the waves were tracked successfully for all acquisitions. Diastole and systole were identified using the ECG signal (Fig. 2a). The average SW speed was 0.6 ± 0.1 m/s at end-diastole (Fig. 2b). The measured speeds ranged from 0.5 ± 0.1 m/s during diastole to 1.9 ± 0.3 m/s during systole. The changes in SW speeds correspond to the expected variation in muscle stiffness during the heart cycle as the RV relaxes and contracts. Conclusions We demonstrate for the first time the induction and tracking of shear waves in the RVFW of a closed-chest pig. The possibility to noninvasively quantify RV wall stiffness opens a large field of translational research, with direct applications in pulmonary hypertension, congenital heart disease and heart failure in general. Pathological increase in stiffness should be further investigated in longitudinal case/control studies. Abstract Figure. Fig1: Induction and tracking of RV SW Abstract Figure. Fig2: SW speeds during cardiac cycle

B-PO02-133 HISTOLOGICAL CHARACTERIZATION OF REVERSIBLE AND IRREVERSIBLE VENTRICULAR LESION BOUNDARIES PRODUCED BY PULSED FIELD ABLATION

During pulsed field ablation (PFA), electric pulses induce reversible (R) or irreversible (IR) injury on cardiac myocytes. To perform histological examination of acute ventricular lesions produced by PFA, determining R and IR injury boundaries in a swine heart model. A 7F catheter with a 3.5mm ablation electrode (TactiCath, Abbott) was connected to a pulsed electric field (PEF) generator (CENTAURI, Galaxy Medical) and positioned in RV and LV under EnSite guidance in 2 closed chest pigs. Biphasic PEF current was delivered between the ablation electrode and a skin patch at 13 RV sites (28Amp, total pulse width of 1.4ms, 4 pulses) and 19 LV sites (35Amp, 1.6ms, 7 pulses). Two hours after ablation, triphenyl tetrazolium chloride (TTC) was administered. Pigs were sacrificed and hearts were excised and fixed in formalin. Hearts were sectioned and stained with hematoxylin and eosin (H&E) and Masson trichrome. Cytochrome c oxidase (COX) staining was also performed to examine mitochondrial activity to delineate R and IR lesion boundaries. Fig. Ablation lesions were well demarcated with TTC staining, showing a dark central zone surrounded by pale boundaries (PB). Histology showed destruction of myocyte architecture within the PB. A hyperstained (dark red) rim beyond the PB indicates R zone. COX staining showed no or low mitochondrial activity within the PB, consistent with IR ablated region. Enhanced activity of COX staining extended to unaffected normal myocardium, consistent with R zone. Acute ventricular lesions produced by PEF ablation show clear demarcation by TTC staining. COX staining suggests that IR lesions are surrounded by R zone.

B-PO03-131 EFFECTS OF CONTACT FORCE ON LESION SIZE DURING PULSED FIELD ABLATION

Effects of contact force (CF) on lesion formation during pulsed field ablation (PFA) have not been validated. To determine the relationship between average CF and lesion size during PFA using a swine beating heart model. A 7F catheter with a 3.5mm saline irrigated (2ml/min) ablation electrode and CF sensor (TactiCath SE, Abbott) was connected to a pulsed electric field (PEF) generator system (CENTAURI, Galaxy Medical). The catheter was positioned in RV and LV under EnSite guidance in 2 closed chest pigs (66kg). During atrial pacing at 90 bpm, biphasic PEF current was delivered between the ablation electrode and a skin patch at 13 separate sites in RV (28Amp, total pulse width of 1.4ms, 4 pulses synchronized with QRS) and 19 separate sites in LV (35Amp, 1.6ms, 7 pulses) at average CF: 1) low (4-12 g, median 8 g); 2) moderate (16-30 g, median 20 g); or 3) high (33-55 g, median 40 g). Ablation was also performed without electrode contact (2 mm away from the endocardium, confirmed by intracardiac echocardiography, n=5). Pigs were sacrificed at 2 hours after ablation and lesion size was measured using triphenyl tetrazolium chloride (TTC) staining. (Fig.) Ablation lesions were clearly identified with TTC staining, showing the dark central zone surrounded by pale boundaries. At constant PEF current and pulse duration, lesion depth increased significantly with increasing contact force. There were no detectable lesions resulting from ablation without electrode contact. At same PEF dose intensity, lesion depth increases significantly with increasing contact force. Electrode-tissue contact is required for effective lesion formation with PEF ablation.

Experimental parameters and infarct size in closed chest pig LAD ischemia reperfusion models; lessons learned

BackgroundPreclinical models that resemble the clinical setting as closely as possible are essential in translating promising therapies for the treatment of acute myocardial infarction. Closed chest pig left anterior descending coronary artery (LAD) ischemia reperfusion (I/R) models are valuable and clinically relevant. Knowledge on the influence of experimental design on infarct size (IS) in these models is a prerequisite for suitable models. To this end, we investigated the impact of several experimental features (occlusion and follow-up time and influence of area at risk (AAR)) on IS.MethodsA total of fifty-one female Landrace pigs were subjected to closed chest LAD balloon occlusion and evaluated in three substudies with varying protocols. To assess the relationship between time of occlusion and the IS, 18 pigs were subjected to 60-, 75- and 90 min of occlusion and terminated after 24 h of follow-up. Influence of prolonged follow-up on IS was studied in 18 pigs after 75 min of occlusion that were terminated at 1, 3 and 7 days. The relation between AAR and IS was studied in 28 pigs after 60 min of occlusion and 24 h of follow-up. The relation between VF, number of shocks and IS was studied in the same 28 pigs after 60 min of occlusion.ResultsIncreasing occlusion time resulted in an increased IS as a ratio of the AAR (IS/AAR). This ranged from 53 ± 23% after 60 min of occlusion to 88 ± 2.2% after 90 min (P = 0.01). Increasing follow-up, from 1 to 3 or 7 days after 75 min of occlusion did not effect IS/AAR. Increasing AAR led to a larger IS/AAR (r2 = 0.34, P = 0.002), earlier VF (r2 = 0.32, P = 0.027) and a higher number of shocks (r2 = 0.29, P = 0.004) in pigs subjected to 60 min of occlusion.ConclusionsThese experiments describe the association of occlusion time, follow-up duration, AAR and VF with IS in closed chest pig LAD I/R models. These results have important implications for future I/R studies in pigs and can serve as a guideline for the selection of appropriate parameters and the optimal experimental design.

The protective effects of phosphodiesterase-5 inhibitor, sildenafil on post-resuscitation cardiac dysfunction of cardiac arrest: by regulating the miR-155-5p and miR-145-5p

BackgroundMiRNA-155 and miRNA-145 have been demonstrated to function as a key regulator in the development of the cardiovascular system. Recent experimental and clinical studies have indicated the cardioprotective role of sildenafil during ischemia/reperfusion (I/R) injury. This study was designed to investigate if administration of sildenafil will attenuate post-resuscitation myocardial dysfunction by regulating miRNA-155 and miR-145 expressions.MethodsThirty-two male pigs (weighing 30 ± 2 kg) were randomly divided into 4 groups, sildenafil group (n = 8), sildenafil +NG-nitro-l-arginine methyl ester (L-NAME) (20 mg/kg L) group (n = 8), saline (SA group, n = 8); and sham operation group (sham group, n = 8). Eight minutes of untreated VF was followed by defibrillation in anesthetized, closed-chest pigs. Hemodynamic status and blood samples were obtained at 0 min, 0.5, 1, 2, 4 and 6 h after return of spontaneous circulation (ROSC), and the hearts were removed and analyzed under electron microscopy, quantitative real-time polymerase chain reaction and ultra structural analysis were performed to evaluate myocardial injury.ResultsCompared with the sildenafil + L-NAME and saline groups, the sildenafil group had better outcomes in terms of hemodynamic and oxygen metabolism parameters as well as 24-h survival rate, and attenuated myocardial injury; In this study, CA pigs showed evidently increased levels of miR-155-5p and miR-145-5p, while the sildenafil treatment decreased the levels of miR-155-5p and miR-145-5p in CA pigs. In addition, the levels of eNOS was decreased in CA pigs, validating sildenafil attenuating post-resuscitation myocardial dysfunction by regulating miRNA-155 and miR-145 expressions.ConclusionsSildenafil group had better outcomes in terms of hemodynamic and oxygen metabolism parameters as well as 24-h survival rate, inhibited the increases in the miR-155-5p and miR-145-5p levels and attenuated myocardial injury in a porcine model of CA and resuscitation.

An intact animal model for the assessment of coronary blood flow regulation "Coronary blood flow regulation".

Coronary blood flow adapts to metabolic demand ("metabolic regulation") and remains relatively constant over a range of pressure changes ("autoregulation"). Coronary metabolic regulation and autoregulation are usually studied separately. We developed an intact animal experimental model to explore both regulatory mechanisms of coronary blood flow. Coronary pressure and flow‐velocities were measured in four anesthetized and closed‐chest pigs using an intracoronary Doppler wire. Metabolic regulation was assessed by coronary flow reserve defined as the ratio between the maximally vasodilated and the basal flow, with hyperemia achieved using intracoronary administration of adenosine (90 µg) or bradykinin (10–6 M) as endothelium‐independent and ‐dependent vasodilators respectively. For both vasodilators, we found a healthy coronary flow reserve ≥ 3.0 at baseline, which was maintained at 2.9 ± 0.2 after a 6‐hr period. Autoregulation was assessed by the lower breakpoint of coronary pressure‐flow relationships, with gradual decrease in coronary pressure through the inflation of an intracoronary balloon. We found a lower limit of autoregulation between 42 and 55 mmHg, which was stable during a 6‐hr period. We conclude that this intact animal model is adequate for the study of pharmacological interventions on the coronary circulation in health and disease, and as such suitable for preclinical drug studies.

Cardiac power output accurately reflects external cardiac work over a wide range of inotropic states in pigs

BackgroundCardiac power output (CPO), derived from the product of cardiac output and mean aortic pressure, is an important yet underexploited parameter for hemodynamic monitoring of critically ill patients in the intensive-care unit (ICU). The conductance catheter-derived pressure-volume loop area reflects left ventricular stroke work (LV SW). Dividing LV SW by time, a measure of LV SW min− 1 is obtained sharing the same unit as CPO (W). We aimed to validate CPO as a marker of LV SW min− 1 under various inotropic states.MethodsWe retrospectively analysed data obtained from experimental studies of the hemodynamic impact of mild hypothermia and hyperthermia on acute heart failure. Fifty-nine anaesthetized and mechanically ventilated closed-chest Landrace pigs (68 ± 1 kg) were instrumented with Swan-Ganz and LV pressure-volume catheters. Data were obtained at body temperatures of 33.0 °C, 38.0 °C and 40.5 °C; before and after: resuscitation, myocardial infarction, endotoxemia, sevoflurane-induced myocardial depression and beta-adrenergic stimulation. We plotted LVSW min− 1 against CPO by linear regression analysis, as well as against the following classical indices of LV function and work: LV ejection fraction (LV EF), rate-pressure product (RPP), triple product (TP), LV maximum pressure (LVPmax) and maximal rate of rise of LVP (LV dP/dtmax).ResultsCPO showed the best correlation with LV SW min− 1 (r2 = 0.89; p < 0.05) while LV EF did not correlate at all (r2 = 0.01; p = 0.259). Further parameters correlated moderately with LV SW min− 1 (LVPmaxr2 = 0.47, RPP r2 = 0.67; and TP r2 = 0.54). LV dP/dtmax correlated worst with LV SW min− 1 (r2 = 0.28).ConclusionCPO reflects external cardiac work over a wide range of inotropic states. These data further support the use of CPO to monitor inotropic interventions in the ICU.

Targeting Mitochondrial Fission Using Mdivi-1 in A Clinically Relevant Large Animal Model of Acute Myocardial Infarction: A Pilot Study.

Background: New treatments are needed to reduce myocardial infarct size (MI) and prevent heart failure (HF) following acute myocardial infarction (AMI), which are the leading causes of death and disability worldwide. Studies in rodent AMI models showed that genetic and pharmacological inhibition of mitochondrial fission, induced by acute ischemia and reperfusion, reduced MI size. Whether targeting mitochondrial fission at the onset of reperfusion is also cardioprotective in a clinically-relevant large animal AMI model remains to be determined. Methods: Adult pigs (30–40 kg) were subjected to closed-chest 90-min left anterior descending artery ischemia followed by 72 h of reperfusion and were randomized to receive an intracoronary bolus of either mdivi-1 (1.2 mg/kg, a small molecule inhibitor of the mitochondrial fission protein, Drp1) or vehicle control, 10-min prior to reperfusion. The left ventricular (LV) size and function were both assessed by transthoracic echocardiography prior to AMI and after 72 h of reperfusion. MI size and the area-at-risk (AAR) were determined using dual staining with Tetrazolium and Evans blue. Heart samples were collected for histological determination of fibrosis and for electron microscopic analysis of mitochondrial morphology. Results: A total of 14 pigs underwent the treatment protocols (eight control and six mdivi-1). Administration of mdivi-1 immediately prior to the onset of reperfusion did not reduce MI size (MI size as % of AAR: Control 49.2 ± 8.6 vs. mdivi-1 50.5 ± 11.4; p = 0.815) or preserve LV systolic function (LV ejection fraction %: Control 67.5 ± 0.4 vs. mdivi-1 59.6 ± 0.6; p = 0.420), when compared to vehicle control. Similarly, there were no differences in mitochondrial morphology or myocardial fibrosis between mdivi-1 and vehicle control groups. Conclusion: Our pilot study has shown that treatment with mdivi-1 (1.2 mg/kg) at the onset of reperfusion did not reduce MI size or preserve LV function in the clinically-relevant closed-chest pig AMI model. A larger study, testing different doses of mdivi-1 or using a more specific Drp1 inhibitor are required to confirm these findings.

How Accurate Is Inverse Electrocardiographic Mapping? A Systematic In Vivo Evaluation.

Inverse electrocardiographic mapping reconstructs cardiac electrical activity from recorded body surface potentials. This noninvasive technique has been used to identify potential ablation targets. Despite this, there has been little systematic evaluation of its reliability. Torso and ventricular epicardial potentials were recorded simultaneously in anesthetized, closed-chest pigs (n=5), during sinus rhythm, epicardial, and endocardial ventricular pacing (70 records in total). Body surface and cardiac electrode positions were determined and registered using magnetic resonance imaging. Epicardial potentials were reconstructed during ventricular activation using experiment-specific magnetic resonance imaging-based thorax models, with homogeneous or inhomogeneous (lungs, skeletal muscle, fat) electrical properties. Coupled finite/boundary element methods and a meshless approach based on the method of fundamental solutions were compared. Inverse mapping underestimated epicardial potentials >2-fold (P<0.0001). Mean correlation coefficients for reconstructed epicardial potential distributions ranged from 0.60±0.08 to 0.64±0.07 across all methods. Epicardial electrograms were recovered with reasonable fidelity at ≈50% of sites (median correlation coefficient, 0.69-0.72), but variation was substantial. General activation spread was reproduced (median correlation coefficient, 0.72-0.78 for activation time maps after spatio-temporal smoothing). Epicardial foci were identified with a median location error ≈16 mm (interquartile range, 9-29 mm). Inverse mapping with meshless method of fundamental solutions was better than with finite/boundary element methods, and the latter were not improved by inclusion of inhomogeneous torso electrical properties. Inverse potential mapping provides useful information on the origin and spread of epicardial activation. However the spatio-temporal variability of recovered electrograms limit resolution and must constrain the accuracy with which arrhythmia circuits can be identified independently using this approach.

In vivo MRI and ex vivo histological assessment of the cardioprotection induced by ischemic preconditioning, postconditioning and remote conditioning in a closed-chest porcine model of reperfused acute myocardial infarction: importance of microvasculature

BackgroundCardioprotective value of ischemic post- (IPostC), remote (RIC) conditioning in acute myocardial infarction (AMI) is unclear in clinical trials. To evaluate cardioprotection, most translational animal studies and clinical trials utilize necrotic tissue referred to the area at risk (AAR) by magnetic resonance imaging (MRI). However, determination of AAR by MRI‚ may not be accurate, since MRI-indices of microvascular damage, i.e., myocardial edema and microvascular obstruction (MVO), may be affected by cardioprotection independently from myocardial necrosis. Therefore, we assessed the effect of IPostC, RIC conditioning and ischemic preconditioning (IPreC; positive control) on myocardial necrosis, edema and MVO in a clinically relevant, closed-chest pig model of AMI.Methods and resultsAcute myocardial infarction was induced by a 90-min balloon occlusion of the left anterior descending coronary artery (LAD) in domestic juvenile female pigs. IPostC (6 × 30 s ischemia/reperfusion after 90-min occlusion) and RIC (4 × 5 min hind limb ischemia/reperfusion during 90-min LAD occlusion) did not reduce myocardial necrosis as assessed by late gadolinium enhancement 3 days after reperfusion and by ex vivo triphenyltetrazolium chloride staining 3 h after reperfusion, however, the positive control, IPreC (3 × 5 min ischemia/reperfusion before 90-min LAD occlusion) did. IPostC and RIC attenuated myocardial edema as measured by cardiac T2-weighted MRI 3 days after reperfusion, however, AAR measured by Evans blue staining was not different among groups, which confirms that myocardial edema is not a measure of AAR, IPostC and IPreC but not RIC decreased MVO.ConclusionWe conclude that IPostC and RIC interventions may protect the coronary microvasculature even without reducing myocardial necrosis.

Influence of acute glycaemic level on measures of myocardial infarction in non-diabetic pigs

>Objective. Patients with diabetes are at increased risk of experiencing myocardial infarction. The influence of the prevailing plasma glucose level on infarction and mortality after acute ischaemia is however unknown. The aim was to study the effect of the acute plasma glucose level on the myocardial infarction size in a closed-chest pig model. Design. 38 non-diabetic pigs were randomised to hypoglycaemic (1.8–2.2 mmol/l; n = 15), normoglycaemic (5–7 mmol/l; n = 12) or hyperglycaemic glucose clamping (22–23 mmol/l; n = 11). After 30 min within glucose target myocardial infarction was induced for 30 min followed by reperfusion for 120 min. Hereafter the heart was double-stained to delineate infarction from viable tissue within the area at risk. Results. Mean infarction size was 201 ± 35 mm2 (mean ± SEM) in the hypoglycaemic group, 154 ± 40 mm2 in the normoglycaemic group and 134 ± 40 mm2 in the hyperglycaemic group, with no differences in infarction size, infarct/area at risk ratio or troponin T levels between the groups. There was no difference in incidence of ventricular fibrillation or mortality between the groups. Conclusion. No statistically significant associations were observed between the acute glycaemic level and measures of myocardial infarction, rates of ventricular fibrillation and subsequent premature death in the setting of acute ischaemia and reperfusion.

Continuous monitoring of cardiac function by 3-dimensional accelerometers in a closed-chest pig model.

Cardiac wall motions reflect systolic and diastolic function. We have previously demonstrated the ability of a miniaturized three-axis (3D) accelerometer to monitor left ventricular function in experimental models and in patients. The main aim of this study was to investigate the clinical utility of the method for monitoring the left and right ventricular function during changes in global and regional cardiac function in a postoperative closed-chest situation. In 13 closed-chest pigs, miniaturized 3D accelerometers were placed on the left ventricle in the apical and basal regions and in the basal region of the right ventricle. An epicardial 3D motion vector was calculated from the acceleration signals in each heart region. Peak systolic velocity along this 3D vector (3D V(sys)) was compared with the positive time derivative of the left and right ventricular pressure and with cardiac index during changes in global LV function (unloading, fluid loading, esmolol, dobutamine) and with ultrasound during regional left ventricular dysfunction (3-min occlusion of the left anterior descending coronary artery). Significant and typical changes in accelerometer 3D V(sys) were seen in all heart regions during changes in global cardiac function. 3D V(sys) reflected the left and right ventricular contractility via significant correlations with the positive time derivative of the left and right ventricular pressure, r = 0.86 and r = 0.72, and with cardiac index r = 0.82 and r = 0.73 (all P < 0.001), respectively. The miniaturized accelerometers also detected regional dysfunction, but showed reduced ability to localize ischaemia as the 3D V(sys) in all heart regions showed similar reductions during coronary artery occlusion. Miniaturized 3D accelerometers placed on the heart can assess global and regional function in a closed-chest model. The technique may be used for continuous postoperative monitoring after cardiac surgery.

Further insights into blood pressure induced premature beats: Transient depolarizations are associated with fast myocardial deformation upon pressure decline

An acute increase in blood pressure is associated with the occurrence of premature ventricular complexes (PVCs). We aimed to study the timing of these PVCs with respect to afterload-induced changes in myocardial deformation in a controlled, preclinically relevant, novel closed-chest pig model. An acute left ventricular (LV) afterload challenge was induced by partial balloon inflation in the descending aorta, lasting 5-10 heartbeats (8 pigs; 396 inflations). Balloon inflation enhanced the reflected wave (augmentation index 30% ± 8% vs 59% ± 6%; P < .001), increasing systolic central blood pressure by 35% ± 4%. This challenge resulted in a more abrupt LV pressure decline, which was delayed beyond ventricular repolarization (rate of pressure decline 0.16 ± 0.01 mm Hg/s vs 0.27 ± 0.04 mm Hg/ms; P < .001 and interval T-wave to peak pressure 1 ± 12 ms vs 36 ± 9 ms; P = .008), during which the velocity of myocardial shortening at the basal septum increased abruptly (ie, postsystolic shortening) (peak strain rate -0.6 ± 0.5 s(-1) vs -2.5 ± 0.8 s(-1); P < .001). It is exactly at this time of LV pressure decline, with increased postsystolic shortening, and not at peak pressure, that PVCs occur (22% of inflations). These PVCs preferentially occurred at the basal and apical segments. In the same regions, monophasic action potentials demonstrated the appearance of delayed afterdepolarization-like transient depolarizations as origin of PVCs. An acute blood pressure increase results in a more abrupt LV pressure decline, which is delayed after ventricular repolarization. This has a profound effect on myocardial mechanics with enhanced postsystolic shortening. Coincidence with induced transient depolarizations and PVCs provides support for the mechanoelectrical origin of pressure-induced premature beats.

The effects of phosphodiesterase-5 inhibitor sildenafil against post-resuscitation myocardial and intestinal microcirculatory dysfunction by attenuating apoptosis and regulating microRNAs expression: essential role of nitric oxide syntheses signaling.

BackgroundRecent experimental and clinical studies have indicated the cardioprotective role of sildenafil during ischemia/reperfusion (I/R) injury. Sildenafil has been shown to attenuate postresuscitation myocardial dysfunction in piget models of ventricular fibrillation. This study was designed to investigate if administration of sildenafil will attenuate post-resuscitation myocardial dysfunction by attenuating apoptosis and regulating miRNA expressions, furthermore, ameliorating the severity of post-microcirculatory dysfunction.MethodsTwenty-four male pigs (weighing 30 ± 2 kg) were randomly divided into groups, sildenafil pretreatment (n = 8), saline (n = 8) and sham operation (sham, n = 8). Sildenafil pretreatment consisted of 0.5 mg/kg sildenafil, administered once intraperitoneally 30 min prior to ventricular fibrillation (VF). Eight minutes of untreated VF was followed by defibrillation in anesthetized, closed-chest pigs. Hemodynamic status and blood samples were obtained at 0 min, 0.5, 1, 2, 4 and 6 h after return of spontaneous circulation (ROSC). Surviving pigs were euthanatized at 24 h after ROSC, and hearts were removed for analysis by electron microscopy, western blotting, quantitative real-time polymerase chain reaction (PCR), and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Intestinal microcirculatory blood flow was visualized by a sidestream dark-field imaging device at baseline and 0.5, 1, 2, 4, and 6 h after ROSC.ResultsCompared with the saline group, the sildenafil group had a higher 24-hour survival (7/8 versus 3/8 survivors, p < 0.05) and a better outcome in hemodynamic parameters. The protective effect of sildenafil also correlated with reduced cardiomyocyte apoptosis, as evidenced by reduced TUNEL-positive cells, increased anti-apoptotic Bcl-2/Bax ratio and inhibited caspase-3 activity in myocardium. Additionally, sildenafil treatment inhibited the increases in the microRNA-1 levels and alleviated the decreases in the microRNA-133a levels which negatively regulates pro-apoptotic genes. At 6 h after ROSC, post-resuscitation perfused vessel density and microcirculatory flow index were significantly lower in the saline group than in the sildenafil group.ConclusionsThe major findings of this study are as follows: (1) sildenafil improved post-resuscitation perfusion of the heart, and thus reduced cardiac myocyte apoptosis and improved cardiac function; (2) sildenafil treatment inhibited the increases in the microRNA-1 levels, but alleviated the decreases in the microRNA-133a levels.

Forward Problem of Electrocardiography

The relationship between epicardial and body surface potentials defines the forward problem of electrocardiography. A robust formulation of the forward problem is instrumental to solving the inverse problem, in which epicardial potentials are computed from known body surface potentials. Here, the accuracy of different forward models has been evaluated experimentally. Body surface and epicardial potentials were recorded simultaneously in anesthetized closed-chest pigs (n=5) during sinus rhythm, and epicardial and endocardial ventricular pacing (65 records in total). Body surface potentials were simulated from epicardial recordings using experiment-specific volume conductor models constructed from magnetic resonance imaging. Results for homogeneous (isotropic electric properties) and inhomogeneous (incorporating lungs, anisotropic skeletal muscle, and subcutaneous fat) forward models were compared with measured body surface potentials. Correlation coefficients were 0.85±0.08 across all animals and activation sequences with no significant difference between homogeneous and inhomogeneous solutions (P=0.85). Despite this, there was considerable variance between simulated and measured body surface potential distributions. Differences between the body surface potential extrema predicted with homogeneous forward models were 55% to 78% greater than observed (P<0.05) and attenuation of potentials adjacent to extrema were 10% to 171% greater (P<0.03). The length and orientation of the vector between potential extrema were also significantly different. Inclusion of inhomogeneous electric properties in the forward model reduced, but did not eliminate these differences. These results demonstrate that homogeneous volume conductor models introduce substantial spatial inaccuracies in forward problem solutions. This probably affects the precision of inverse reconstructions of cardiac potentials, in which this assumption is made.