Gordon H. Templeton

Reduced myocardial reflow and increased coronary vascular resistance following prolonged myocardial ischemia in the dog.

Studies were performed to determine whether an alteration in coronary vascular resistance and a reduction in the reflow phenomenon occurred in the blood-perfused, heparinized canine heart after various periods of myocardial ischemia. Regional myocardial blood flow was measured with radioactive microspheres. Proximal left anterior descending coronary artery blood flow was measured with a periarterial flow transducer. Reduced reflow to the ischemic portion of the left ventricle and increased resistance in the left anterior descending coronary artery were present after 120 minutes of myocardial ischemia. The reduction in reflow was specific to the subendocardium of the ischemic area. Saline and isosorbide dinitrate (Isordil) did not prevent the increase in coronary vascular resistance or the significant reduction in reflow to the subendocardial portion of the ischemic area. Hypertonic mannitol given so as to increase serum osmolality 40 mosmoles/kg prevented the increase in coronary vascular resistance and modified the reduction in the reflow phenomenon to the subendocardial portion of the ischemic area. Thus, both an increase in coronary vascular resistance and a significant reduction in reflow to the subendocardial portion of the ischemic area occur in the canine heart after 120 minutes of myocardial ischemia. Moreover, the increase in coronary vascular resistance can be prevented and the reduction in reflow to the subendocardial portion of the ischemic area can be modified by the administration of hypertonic mannitol.

Relationship between Epicardial S-T Segment Changes and Myocardial Metabolism during Acute Coronary Insufficiency

Canine left ventricular wall metabolites (ATP, creatine phosphate, glycogen, glucose, glucose-6-phosphate, and lactate) were assessed in the ischemic region of the wall after 17 minutes of ligation of the left anterior descending coronary artery at its midlevel. Variations in the degree of lactate accumulation were found between different sites in the same ischemic region and between the outer and the inner left ventricular wall at the same site. High levels of lactate were associated with S-T segment elevation in the epicardial electrocardiogram, but no lactate accumulation or only mild lactate accumulation was found at isoelectric sites. Lactate accumulation at isoelectric sites was higher in the outer wall than it was in the inner wall; the opposite tendency was observed at sites of S-T segment elevation. In addition to high lactate levels, sites of S-T segment elevation demonstrated a more pronounced depletion of ATP and creatine phosphate, indicating not only a marked anaerobic glycolysis but also a more pronounced overall anaerobic stress. No relationship was found, however, between the absolute magnitude of S-T segment elevation and the degree of lactate accumulation or ATP depletion at sites of S-T segment elevation within the ischemic region. The data obtained in this study demonstrate that during myocardial ischemia epicardial sites of S-T segment elevation are sites of pronounced subendocardial and epicardial ischemia and of anaerobic metabolism.

Influence of aging on left ventricular hemodynamics and stiffness in beagles.

We studied the influence of aging on the contractile performance, stiffness, and contraction time of the canine left ventricle. Eight young (27 ± 2.5 months, mean ± SE) and seven old (128 ± 20.5 months) beagles were placed on complete cardiopulmonary bypass, the arterial pressure was adjusted to 80 mm Hg, and the heart contracted isovolumically at a paced rate of 120 beats/min. Diastolic pressure-volume curves were established for each unpaced left ventricle at the beginning of each experiment, and the volume at the knee of the curve was used during the subsequent data collection when the heart was paced. Stiffness was measured with a sinusoidal forcing function, which imposed a sinusoidal volume displacement of 1 ml at 20 Hz into a balloon placed in the left ventricle. In each ventricle, stiffness was related linearly to pressure during the cardiac cycle, and was greater for any given pressure in the older beagles. Contraction duration was prolonged in the older dogs. In an additional seven old beagles during right heart bypass, time and duration of contraction were longer than in seven young beagles. Aging of the beagle heart is associated with an increase in left ventricular systolic and diastolic stiffness and prolonged duration of contraction. Circ Res 44: 189-194, 1979

Lanthanum Probe Studies of Cellular Pathophysiology Induced by Hypoxia in Isolated Cardiac Muscle

This study was undertaken to evaluate directly the relationship between evolution of irreversible myocardial injury induced by hypoxia in an isolated papillary muscle preparation and the development of pathophysiological alterations related to severely impaired membrane function. An ionic lanthanum probe technique was employed as a cytochemical marker to monitor the progression of cellular injury, and data from this cytologic technique were correlated with ultrastructure and measurements of contractile parameters in a total of 67 muscles subjected to control conditions or to graded intervals of hypoxia with or without reoxygenation. Marked depression of developed tension and rate of tension development occurred after 30 min of hypoxia. Contractile function showed significant recovery with reoxygenation after 1 h and 15 min of hypoxia but remained depressed when reoxygenation was provided after 2 or 3 h of hypoxia. Examination by transmission and analytical electron microscopy (energy dispersive X-ray microanalysis) revealed lanthanum deposition only in extracellular regions of control muscles and muscles subjected to 30 min of hypoxia. After hypoxic intervals of over 1 h, abnormal intracytoplasmic and intramitochondrial localization of lanthanum were detected. After 1 h and 15 min of hypoxia, abnormal intracellular lanthanum accumulation was associated with only minimal ultrastructural evidence of injury; muscle provided reoxygenation after 1 h and 15 min of hypoxia showed improved ultrastructure and did not exhibit intracellular lanthanum deposits upon exposure to lanthanum during the reoxygenation period. After 2 to 3 h of hypoxia, abnormal intracellular lanthanum accumulation was associated with ultrastructural evidence of severe muscle injury which persisted after reoxygenation. Thus, the data support the conclusion that cellular and membrane alterations responsible for abnormal intracellular lanthanum deposition precede the development of irreversible injury but evolve at a transitional stage in the progression from reversible to irreversible injury induced by hypoxia in isolated feline papillary muscles.

Influence of Temperature on the Mechanical Properties of Cardiac Muscle

The influence of temperature on contractile performance, total myocardial stiffness, and the elastic and viscous components of stiffness was studied in intact hearts (dog) and isolated cardiac muscle (cat). In 12 dogs on whole-heart bypass, stiffness (ΔP/ΔV) was determined by inducing sinusoidal 0.5-ml volume changes in paced, isovolumically contracting left ventricles and measuring the resultant pressure changes (ΔP). In eight cat papillary muscles contracting isometrically at Lmax, stiffness (ΔT/ΔL) was similarly determined from the changes in tension and length during sinusoidal length changes of 0.25% L max. Total stiffness was linearly related to ventricular pressure or muscular tension during both contraction and rest. As the temperature rose from 33° to 40°C in vivo and from 22° to 40°C in vitro, the slopes of the stiffness-pressure and stiffness-tension relationships and the intercept of the latter declined; therefore, stiffness is inversely related to temperature. Since both preparations responded as linear second-order systems, stiffness could be separated into its elastic and viscous components. The moduli of both components varied in the same direction as total stiffness during temperature changes; however, the viscous component was more sensitive to the influence of temperature.

Influence of acute myocardial depression on left ventricular stiffness and its elastic and viscous components.

The influence of acute myocardial depression on ventricular stiffness and on its elastic and viscous components was studied in 19 dogs. After the animals were placed on cardiopulmonary bypass, stiffness was measured by sinusoidally injecting volume changes of 0.5 ml (deltaV) at 22 Hz into paced, isovolumically (deltaP) of the sinusoidal pressure response. Stiffness was linearly related to pressure (P) throughout the cardiac cycle, so that deltaP/delta V = alpha P + beta, where alpha and beta are constants. Myocardial depression was induced in one of three different ways: by coronary artery ligation, by administration of propranolol (Inderal), or by administration of pentobarbital. All three interventions caused significant increases in the slope, alpha, of the stiffness-pressure relationship, while the intercept, beta, remained unchanged. Release of the coronary occlusion or administration of acetylstrophantidin partially reversed depression and the change in alpha; Approximation of the mechanical nature of the left ventricle in terms of a linear second-order mechanical system permitted the division of stiffness into its elastic and viscous components. Like total stiffness, both the elastic and the viscous components were linearly related to ventricular pressure. Elastic stiffness was not changed, but the slope of the line relating viscous stiffness to pressure was significantly increased during ischemic depression, indicating that a change in viscosity was primarily responsible for the increase in total ventricular stiffness.

The influence of hypertonic mannitol on regional myocardial blood flow during acute and chronic myocardial ischemia in anesthetized and awake intact dogs.

The influence of hypertonic mannitol on regional myocardial blood flow and ventricular performance was studied during acute myocardial ischemia in awake, unsedated and in anesthesized dogs and after myocardial infarction in awake unsedated dogs. Regional myocardial blood flow was measured with radioactive microspheres. Generalized increases in regional myocardial blood flow occurred after mannitol in all of the different animal models studied. The increases in coronary blood flow after mannitol were just as impressive in the nonischemic regions as in the ischemic portion of the left ventricle in all of the different models that were examined in this study. Improvement in regional myocardial blood flow to the ischemic area of the left ventricle after mannitol was associated with a reduction in ST segment elevation during acute myocardial ischemia in anesthetized dogs. The increases in regional myocardial flow after mannitol were also associated with increases in contractility, but the increases in flow appeared to be more impressive than the changes in contractility. The data obtained demonstrate that mannitol increases regional coronary blood flow to both ischemic and nonischemic myocardium in both anesthetized and awake, unsedated, intact dogs with acute and chronic myocardial ischemia and that mannitol reduces ST segment elevation during acute myocardial ischemia in anesthetized dogs. Thus the results suggest that under these circumstances the increases in regional myocardial blood flow after mannitol are of physiological importance in reducing the extent of myocardial injury. Since coronary blood flow increased to nonischemic regions the increases in regional myocardial flow demonstrated in this study after mannitol cannot be entirely explained by the mechanism of reduction in ischemic cell swelling.

Influence of Calcium on the Inotropic Actions of Hyperosmotic Agents, Norepinephrine, Paired Electrical Stimulation, and Treppe

To analyze the interaction of calcium ion concentration with hypertonic agents and with other inotropic interventions, isolated right ventricular cat papillary muscles were studied under isometric conditions in Krebs-Ringer bicarbonate solution. Extracellular calcium concentrations were varied between 2.5 and 11.0 mM. Maximal inotropic effects occurred between 5 and 8.0 mM calcium and further elevation to 11.0 mM was without additional influence. The effect of hyperosmotic sucrose and mannitol on papillary muscle performance was compared with that of 10(-6) M norepinephrine at calcium concentrations of 2.5 and 10.0 mM and with paired electrical stimulation in 10.0 mM calcium. Both norepinephrine and the hyperosmotic agents produced significant increases in developed tension and in the maximal rate of tension rise (dT/dt) in Krebs-Ringer in 2.5 and 4.0 mM calcium. In 10 mM calcium norepinephrine increased developed tension and dT/dt, but sucrose and mannitol caused no change or small reductions in both. Paired electrical stimulation, like hyperosmolality, caused no increase in dT/dt in 10 mM calcium. The presence of a potent pharmacological inhibitor of systolic calcium transfer across the cell membrane (D600, 10(-6) M) reduced developed tension and dT/dt by 76+/-2.7 and 74+/-2.0%, respectively, and prevented and in fact reversed the expected increase in dT/dt associated with an increase in rate of stimulation (treppe). However, hypertonic mannitol and paired pacing persisted in causing marked increases in developed tension and dT/dt even in the presence of D600, suggesting that their inotropic effects are not dependent on increased intracellular transfer of calcium during systole through cell membrane channels in which D600 acts as a competitive inhibitor. The results of these studies suggest that apparent functional saturation of intracellular calcium receptor sites eliminates any additional inotropic effect of hyperosmolality or paired pacing. The data are compatible with the hypothesis that the inotropic effects of hyperosmolality and of paired pacing result from an increase in calcium concentration at the myofilaments during contraction. The increase induced by hyperosmolality might occur because of an increase in the total amount of calcium released into the cytosol with each action potential and/or as a passive consequence of cellular dehydration. Norepinephrine has the capacity to increase contractility even when intracellular calcium receptor sites appear to be functionally saturated, suggesting that it may act at least in part by a mechanism that is independent of changes in net intracellular calcium concentration.