Dipti P. Rath

Sodium Pentobarbital Versus α-Chloralose Anesthesia Experimental Production of Substantially Different Slopes in the Transmural CP/ATP Ratios Within the Left Ventricle of the Canine Myocardium

BACKGROUND Transmural analyses of the creatine phosphate (CP)/ATP ratio in various lamina of the canine myocardium have previously revealed significant variations in the CP/ATP ratio, with the subendocardial layer displaying a decreased ratio relative to the subepicardial layer. Without exception, these results were obtained under sodium pentobarbital anesthesia. These findings have been interpreted to imply that the normal endocardium may be operating in the oxygen-limited domain or that there are transmurally varying set points for the regulation of oxidative phosphorylation. METHODS AND RESULTS In this work, we examine the effect of the anesthetic regimen on the transmural CP/ATP ratio within the left ventricular wall of the canine myocardium using spatially localized 31P-nuclear magnetic resonance (NMR) and an open-chest model. Two anesthetics were compared, alpha-chloralose and sodium pentobarbital. Under sodium pentobarbital, the CP/ATP ratio ranged from 1.92 +/- 0.06 to 2.51 +/- 0.08 from endocardium to epicardium, resulting in a transmural slope in the CP/ATP ratio of 0.149 +/- 0.047 (n = 22). Under alpha-chloralose, CP/ATP ratios ranged from 2.18 +/- 0.05 to 2.32 +/- 0.06, with a transmural slope of 0.035 +/- 0.018 (n = 38). Thus, the transmural slope in CP/ATP ratio was nearly four times greater with sodium pentobarbital than with alpha-chloralose, and the difference in these slopes was statistically significant (P = .029). No difference was observed in average CP/ATP obtained from the entire wall with either anesthetic. CONCLUSIONS These results demonstrate that the transmural trend in CP/ATP ratio previously reported in the myocardium is likely to be a direct reflection of the sodium pentobarbital anesthetic regimen, not truly reflecting the trend in the normal unanesthetized animal. Moreover, since the transmural variation in CP/ATP ratio was greatly reduced with alpha-chloralose, it appears unlikely that the endocardium in the normal unanesthetized heart is operating in the oxygen-limited domain. These results also point to the importance of the anesthetic regimen in biochemical analysis, indicate the necessity of increased caution in directly translating results obtained under anesthesia, and demonstrate the unique power of in vivo NMR to extract such subtle biochemical information.

Flow Requirements in Ventricular Fibrillation: An In Vivo Nuclear Magnetic Resonance Analysis of the Left Ventricular High-Energy Phosphate Pool☆☆☆★★★

STUDY OBJECTIVE We sought to determine whether flow rates of approximately 60% of normal values are sufficient to preserve the left ventricular myocardial high-energy phosphate pool during ventricular fibrillation (VF). METHODS Mixed-breed swine (weight 22. 4+/-2.5 kg) were anesthetized with alpha-chloralose, placed in a state of VF, and perfused with extracorporeal circulation at a target flow of 50 mL.kg(-1).min(-1). In vivo whole-wall (average of left ventricular wall) and spatially localized phosphorous-31 nuclear magnetic resonance (NMR) spectra were acquired at baseline and during VF. RESULTS Mean flow during VF was 58+/-20 mL.kg(-1). min(-1) (+/-SD; 95% confidence interval, 44 to 71) or about 60% of baseline cardiac output (n=13). Whole-wall adenosine triphosphate (ATP) decreased during perfused VF (P <.05), whereas creatine phosphate (CP) remained unchanged from baseline. With spatially localized NMR, the ratios of CP/ATP were similar at baseline in all layers (endocardium --> epicardium) of the left ventricular wall. However, during perfused VF, subepicardial CP/ATP ratios increased by 14% to 40% compared with baseline values, whereas subendocardial CP/ATP ratios remained unchanged (1% to 3% increase). An additional 4 animals perfused at 72+/-10 mL.kg(-1).min(-1) (+/-SD; 95% confidence interval, 56 to 92) during VF had preservation of CP and ATP levels. CONCLUSION Flow levels equivalent to 60% of baseline cardiac output were insufficient to maintain normal high-energy phosphate levels in the in vivo fibrillating myocardium. At this level of flow, myocardial high-energy phosphate loss is nonhomogeneous within the left ventricular wall.

Left ventricular myocardial adenosine triphosphate changes during reperfusion of ventricular fibrillation: the influence of flow and epinephrine.

Objective: To determine whether epinephrine in combination with high flow worsens left ventricular (LV) myocardial high‐energy phosphate stores during reperfusion of ischemic ventricular fibrillation (VF). Design: Blinded, prospective block randomized, placebo controlled study. Setting: University medical center research laboratory. Subjects: A total of 22 mixed breed swine weighing 22.0 ± 3.3 kg (SD). Interventions: Open‐chest swine, anesthetized with α‐chloralose, underwent 10 mins of nonperfused VF followed by reperfusion with cardiopulmonary bypass for 90 mins and then defibrillation. Animals were block randomized to four groups for reperfusion: Group 1 (n = 5), high flow (100 mL/kg/min) and epinephrine (2.5 μg/kg/min); Group 2 (n = 5), high flow and placebo; Group 3 (n = 6), low flow (30 mL/kg/min) and epinephrine; and Group 4 (n = 6), low flow and placebo. Measurements and Main Results: In vivo LV creatine phosphate (CP) and adenosine triphosphate (ATP) were determined using whole wall and spatially localized 31P NMR spectroscopy at 4.7 Tesla. During perfusion of the fibrillating myocardium, epinephrine significantly increased aortic pressure (p < .05) and improved defibrillation rates (p < .01). ATP levels during reperfusion were significantly decreased within all groups compared with baseline. There were no differences in ATP levels between groups. High flow, independent of epinephrine, was associated with increased preservation of ATP (p < .05), increased CP/ATP ratios (p < .02) in all layers of the LV wall, and decreased aortic and cardiac vein lactates (p < .001). Conclusions: Epinephrine, in combination with flow higher than standard cardiopulmonary resuscitation flows, increased perfusion pressure and defibrillation rates, but did not significantly alter myocardial ATP during VF reperfusion in the in vivo heart. Reperfusion flow, independent of epinephrine, is a critical determinant of myocardial ATP preservation.

Myocardial metabolic changes during reperfusion of ventricular fibrillation : a 31P nuclear magnetic resonance study in swine

OBJECTIVE Myocardial metabolic requirements during reperfusion of ventricular fibrillation are poorly understood. The objective of this study was to determine if controlled reperfusion after a clinically relevant global ischemia period of 10 mins was sufficient to prevent or reverse myocardial ischemia as indicated by changes in myocardial high energy phosphates, myocardial intracellular pH, and great cardiac vein lactate. DESIGN Prospective laboratory study with controlled reperfusion. SETTING Research laboratory at a university medical center. SUBJECTS Five swine weighing 19 +/- 3 kg. INTERVENTIONS Ten minutes of nonperfused ventricular fibrillation followed by reperfusion with cardiopulmonary bypass (flow 30 mL/kg/min) for 50 mins. MEASUREMENTS AND MAIN RESULTS Myocardial adenosine triphosphate (ATP), phosphocreatine, and intracellular pH were determined using in vivo 31P nuclear magnetic resonance. Myocardial blood flow, measured by 15-mu radiolabeled microspheres, was significantly increased above baseline during reperfusion. Phosphocreatine was depleted during the 10 mins of nonperfused ventricular fibrillation, but recovered to 122 +/- 18% of baseline with reperfusion and was 112 +/- 18% at 60 mins (p < .005). ATP concentrations decreased to 51 +/- 16% of baseline after 10 mins of nonperfused ventricular fibrillation, improved to 67 +/- 9% of baseline with early reperfusion, and were 65 +/- 9% of baseline at 60 mins (p < .02). Myocardial intracellular pH improved from 6.11 +/- 0.18 after 10 mins of nonperfused ventricular fibrillation, to 6.89 +/- 0.20 with early reperfusion, and then decreased to 6.85 +/- 0.35 at 60 mins ventricular fibrillation (p < .001). Despite myocardial blood flows higher than baseline during the reperfusion period, great cardiac vein/aortic lactate gradient increased over the reperfusion period. CONCLUSION Prolonged reperfusion with supranormal myocardial blood flow does not restore normal myocardial aerobic metabolism in the fibrillating myocardium after a 10-min nonperfused ventricular fibrillation period.

Spatial localization with modified Fourier series windows. Application to the transmural 13C-nuclear magnetic resonance analysis of the in vivo myocardium.

RATIONALE AND OBJECTIVES A modified Fourier Series Window (FSW) method is introduced that provides a simple, reasonably accurate, solution to spatial localization for layers in nuclear magnetic resonance (NMR) spectroscopy. This method was developed because signal originating from spins immediately adjacent to the coil plane can leak into standard FSW localized spectra obtained from more distal layers. METHODS The B1 profile of the surface coil and a priori sample knowledge were used to generate modified FSW coefficients that largely compensate for contamination from proximal layers. These modified coefficients were used to acquire spatially localized spectra from a three-layered phantom containing inorganic phosphate, phosphocreatine, and pyrophosphate. Spatially localized spectra also were obtained from the open-chest canine myocardium. RESULTS The modified Fourier Series Window approach was validated using a quality assurance phantom. This method was then applied to the transmural analysis of 13C-containing metabolites in the in vivo canine myocardium during 3-(13)C sodium pyruvate infusion (n = 10). In vivo NMR spectra were characterized by resonances corresponding to the C2, C3, and C4 carbons of glutamic acid and to endogenous triacyglycerides and surface lipids. A transmural trend in metabolites could be observed under these conditions. This trend, however, was likely to result from the interference of surface lipids in the epicardial layer. CONCLUSIONS The authors demonstrate that the modified Fourier Series Window method can be applied in situations where signal-to-noise considerations limit the applicability of more sophisticated spatial localization methods. In addition, the authors report a slight gradient in the endogenous triacylglyceride resonance, which is likely to originate from the presence of surface lipids.