Cardiac function and glycogen content after twenty-four-hour preservation with various metabolic substrates.

BACKGROUND: Previous work has suggested that recovery of cardiac function after long preservation periods is improved if a suitable exogenous substrate is provided. However, interpretation of existing data on the benefit of added substrate is complicated by differing preservation media and protocols studied, as well as differing models used to evaluate postpreservation recovery. These experiments were designed to evaluate glucose, pyruvate, aspartate, and glutamate as exogenous substrates for prolonged perfusion preservation of hearts with a rat heart model preserved with crystalloid medium and with function quantified in an isolated working heart preparation. METHODS: Cardiac function (n = 5/group) and tissue glycogen content (n = 5/group) were measured in fresh control rat hearts and four groups of hearts preserved for 24 hours in an extracellular-type cardioplegic medium containing 11 mmol/L glucose and either 20 mmol/L sodium aspartate, 20 mmol/L sodium glutamate, 20 mmol/L sodium pyruvate, or no other substrate. Postpreservation cardiac function was measured in an isolated working rat heart preparation for a 4-hour reperfusion period. Exogenous substrate consumption during preservation and tissue glycogen content at the end of preservation were measured with spectrophotometric assays. RESULTS: All hearts in the aspartate- and glutamate-preserved groups functioned for the full 4-hour period with stroke work that was 50% to 60% of control. Hearts preserved with only glucose substrate had inconsistent recovery: two of five hearts in that group did not recover, whereas three recovered similar to the aspartate and glutamate groups. None of the pyruvate-preserved hearts recovered contractile function. There was no association between postpreservation tissue glycogen content and extent of cardiac function recovery. CONCLUSION: Aspartate or glutamate plus glucose was the best substrate mixture among those tested for long-term perfusion preservation of hearts. These amino acids offer advantages over pyruvate because they produced better recovery of the hearts and because they are chemically more stable than pyruvate. Aspartate or glutamate plus glucose also produced better postpreservation function compared with glucose alone.

Recovery of sheep hearts after perfusion preservation or static storage with crystalloid media.

BACKGROUND: These experiments were designed to evaluate the viability of large hearts after preservation by use of procedures that have shown good results with small animal hearts. Efficacy of novel long-term preservation protocols should be documented with a large animal model before such procedures can be adopted for clinical use. We studied the recovery of sheep hearts that were perfusion-preserved in media containing two different substrate mixtures and hearts stored without perfusion either in University of Wisconsin solution modified to maintain tissue adenosine triphosphate content or in Stanford solution. METHODS: Six groups of sheep hearts were studied: I, fresh nonpreserved controls; II, hearts perfusion-preserved at 11 degrees C for 24 hours by use of an oxygenated extracellular-type medium with pyruvate + glucose substrate; III, hearts preserved as for II but with aspartate + glutamate + glucose substrate; IV, hearts stored without perfusion at 3 degrees C for 24 hours in University of Wisconsin solution containing 2,3-butanedione monoxime 30 mmol/L, CaCl2 1 mmol/L, and fresh reduced glutathione 3 mmol/L; V, hearts stored without perfusion at 3 degrees C for 4 hours in Stanford solution; VI, hearts preserved as for II but without perfusion. Recovery was measured for 6 hours in a Langendorff model, perfused with an erythrocyte + albumin medium. RESULTS: Hearts that were perfusion-preserved with both substrate mixtures and hearts stored in modified University of Wisconsin solution recovered function that was not significantly different from control subjects. Hearts stored in Stanford medium did not recover as well as did groups II, III, and IV. Left ventricular pressure and peak rate of left ventricular relaxation of the Stanford group were lower, and left ventricular enddiastolic pressure was higher, than those values for controls (repeated measures analysis of variance; Dunnett's procedure). The group VI hearts did not recover function at all. CONCLUSION: The results suggest that large hearts preserved with medium containing either aspartate + glutamate + glucose or pyruvate + glucose have comparable recovery after long-term perfusion preservation. Aspartate + glutamate may offer advantages for clinical use because of their lower production of lactate and better chemical stability compared with pyruvate. Static storage in modified University of Wisconsin solution also produced viable hearts with recovery comparable to perfusion-preserved aspartate + glutamate + glucose hearts. Tests of these preservation media and procedures with large transplanted hearts are warranted.

Steroid pretreatment improves graft recovery in a sheep orthotopic heart transplantation model.

BACKGROUND: Previous reports provide conflicting evidence concerning effects of steroids on recovery of cardiac function during procedures involving cardiopulmonary bypass. This study was designed to test the hypothesis that pretreatment of animals with steroids before heart transplantation improves graft hemodynamic function. METHODS: Four groups of sheep were studied: CON, nonsteroid-treated nontransplanted controls (n = 8); CON-S, steroid-treated nontransplanted controls (n = 5); TX, nonsteroid-treated transplanted animals (n = 5); and TX-S, steroid-treated transplanted animals (n = 5). Steroid-treated animals were given methylprednisolone 30 mg/kg immediately before surgery. Procedures for harvest and orthotopic transplantation were similar to those used clinically. Contractile function, left ventricular diameter, and cardiac output of control and transplanted hearts were measured for 6 hours. A 2 x 2 factorial repeated measures analysis of variance was used to determine statistical significance (p < 0.05). RESULTS: Steroid pretreatment produced significantly higher function in controls and transplanted animals compared with nonsteroid-treated animals. On average over 6 hours, significant steroid effects were observed for left ventricular peak systolic pressure, mm Hg (CON, 85 +/- 2; CON-S, 98 +/- 3; TX, 74 +/- 3; TX-S, 91 +/- 2); global stroke work, mJoule x cm(-2) (CON, 4.69 +/- 0.21; CON-S, 5.88 +/- 0.32; TX, 2.27 +/- 0.17; TX-S, 4.23 +/- 0.16); and peak rate of pressure relaxation (-dP/dt(max)), mm Hg/msec (CON, 1.23 +/- 0.05; CON-S, 1.44 +/- 0.08; TX, 0.60 +/- 0.03; TX-S, 2.04 +/- 0.13). Steroid pretreatment produced more stable recovery for transplanted animals. All five TX-S animals could be removed from inotropic support and had stable function for 6 hours. In contrast, 1 of 5 TX animals could not be removed from inotropic support, and 1 of 5 TX hearts failed 3 hours after transplant. Arterial blood PO2 values were significantly higher in steroid-treated animals than in nonsteroid treated animals. Blood systemic lactate, which was elevated after transplantation, returned to control level by 6 hours in the steroid-treated group but not in the nonsteroid-treated group. CONCLUSION: Steroid pretreatment of heart donors and recipients improved systolic and diastolic function and hemodynamic stability after transplantation. In addition, steroid pretreatment improved pulmonary gas exchange of control and transplanted animals.

Incomplete recovery of working heart function after twenty-four-hour preservation with a modified University of Wisconsin solution.

BACKGROUND AND METHODS: This study was designed to determine the function of isolated rabbit hearts after static preservation with modified University of Wisconsin solution for 24 hours. Commercially available University of Wisconsin solution, modified with CaCl2 1 mmol/L and 2,3-butanedione monoxime 30 mmol/L, was used as the preservative. After flushing the coronary vasculature with medium, hearts were submersion stored at 1 degree C to 4 degrees C. After preservation, isolated heart function at 37 degrees C was quantified for 30 minutes in a non-ejecting mode and for 4 hours ejecting at a physiologic workload. Fresh control hearts (n = 5) and University of Wisconsin solution-preserved hearts (n = 6) were studied. RESULTS: Nonworking (non-ejecting) left ventricular function of the two groups did not differ, except for peak rate of left ventricular pressure development which was higher for the University of Wisconsin solution hearts than for controls. When the hearts were subjected to a physiologic workload, however, left ventricular function of the two groups differed significantly. Three of the six University of Wisconsin solution hearts failed before the 4-hour perfusion end point, whereas all five control hearts maintained stable working function for the full 4 hours. The University of Wisconsin solution hearts, while in the ejecting mode, exhibited significantly impaired function. Mean values were as follows (p < 0.05): left ventricular systolic pressure (in millimeters of mercury), control 105 +/- 1, University of Wisconsin solution 86 +/- 4; peak rate of left ventricular pressure development (in millimeters of mercury per millisecond), control 3.33 +/- 0.11, University of Wisconsin solution 2.39 +/- 0.24; cardiac output (in milliliters per minute per gram), control 400 +/- 25, University of Wisconsin solution 288 +/- 26; stroke work (in milliJoules per gram), control 20.1 +/- 1.3, University of Wisconsin solution 11.9 +/- 1.1; left ventricular end-diastolic pressure (in millimeters of mercury), control 5.4 +/- 0.3, University of Wisconsin solution 10.2 +/- 1.3; peak aortic flow rate (in milliliters per minute), control 946 +/- 9, University of Wisconsin solution 659 +/- 44; millimoles of lactate produced in 30 min/Joule stroke work, control 0.50 +/- 0.06, University of Wisconsin solution 6.99 +/- 0.37. CONCLUSIONS: These results indicate that (1) hypothermic storage in this modified University of Wisconsin solution does not preserve hearts sufficiently to support a physiologic workload for an extended period and (2) assessment of post-preservation function with a non-ejecting heart model does not accurately predict the ability of the preserved heart to support a physiologic workload.

Posttransplantation function of hearts preserved with fluorochemical emulsion.

This study was designed to determine whether the novel perfluoroperhydrophenanthrene-egg yolk phospholipid emulsion, APE-LM, was an effective oxygen carrier for long-term hypothermic heart preservation. We postulated that hearts preserved with APE-LM would be well oxygenated during 24-hour preservation and that reperfusion of such hearts with blood would not produce functional or metabolic evidence of myocardial ischemia. Four groups of rabbit hearts were studied (n = 7 per group): fresh controls: nonpreserved, nontransplanted hearts; surgical controls: fresh hearts transplanted heterotopically for 75 minutes before explant and study for 4 hours as isolated working hearts perfused at 37 degrees C; crystalloid-preserved: hearts preserved with crystalloid medium, followed by transplantation and isolated heart perfusion; APE-LM-preserved: hearts treated as those in the crystalloid-preserved group, but preservation was with medium containing APE-LM emulsion (10 ml/dl). Preservation was with continuous coronary perfusion at 18 mm Hg pressure, 12 degrees C, and oxygen tension 838 +/- 11 mm Hg. During preservation, APE-LM hearts had significantly higher pyruvate consumption, and correspondingly higher oxygen consumption, than that of crystalloid hearts. No significant differences were found among fresh controls, surgical controls, and APE-LM-preserved hearts with respect to contractile or output function, oxygen consumption and efficiency indexes, or lactate production during in vitro perfusion. Left ventricular peak systolic pressure and peak rate of pressure development were significantly lower for crystalloid-preserved hearts than for fresh and surgical controls. Left ventricular end-diastolic pressure of crystalloid-preserved hearts was higher than that of the other three groups. The data indicate that rabbit hearts in this model were well preserved with APE-LM and that this emulsion produced better recovery of function than did crystalloid preservation, possibly as a consequence of the high oxygen delivery by the fluorocarbon during preservation.

Long-term heart preservation by intermittent perfusion with crystalloid medium.

This study was undertaken to determine whether hearts preserved with intermittent coronary perfusion would recover physiologic function after a prolonged period of hypothermic preservation. Intermittent perfusion is commonly used for cardioplegia, but its efficacy in long-term heart preservation has not yet been demonstrated. Five groups of isolated rat hearts were studied (n = 7 per group): (1) fresh nonpreserved control hearts; (2) hearts preserved with continuous low-pressure perfusion via the aorta; (3) hearts preserved with cycles of 5 minutes of perfusion followed by 25 minutes of nonperfusion; (4) hearts preserved with cycles of 10 minutes of perfusion followed by 25 minutes of nonperfusion; (5) hearts preserved with submersion storage without perfusion. An oxygenated extracellular-type crystalloid medium (oxygen tension = 820 +/- 5 mm Hg) was used as a preservation medium; preservation was for 12 hours. During preservation, the coronary resistance of the intermittent perfusion-preserved hearts increased significantly, and these hearts produced significantly more excess lactate than did hearts in the other two preservation groups. The submersion-stored hearts exhibited no postpreservation ventricular function in an isolated perfused working rat heart system. The poststorage function of the other four groups, which was quantified during a 4-hour, 37 degrees C perfusion period at constant heart rate, indicated that there were no significant group differences with respect to output or energetics (coronary flow, aortic output, cardiac output, myocardial oxygen consumption, and external work efficiency). The intermittent perfusion-preserved hearts had significantly lower postpreservation contractile function (left ventricular systolic pressure, peak rates of left ventricular pressure development and relaxation, peak aortic flow rate, stroke work, and peak power) and higher left ventricular end-diastolic pressure compared with the control group. Although hearts preserved with intermittent perfusion had a loss of contractile function and decreased compliance compared with fresh hearts, after preservation they had better function than did hearts preserved with submersion storage and the same function as hearts preserved with continuous perfusion.

Importance of substrate enhancement for long-term heart preservation.

These experiments tested the hypothesis that addition of pyruvate to a preservation medium would improve postpreservation cardiac function as quantified in an isolated working heart model after heterotopic transplantation. Four groups of rabbit hearts were studied (n = 5 per group): fresh controls, fresh hearts perfused as isolated working hearts; surgical controls, fresh hearts transplanted heterotopically and reperfused with blood for 75 minutes before being studied as isolated hearts; preserved without pyruvate, hearts perfusion-preserved with oxygenated extracellular-type crystalloid medium; preserved with pyruvate, same same as the group without pyruvate, but medium contained 20 mmol/L pyruvate. After preservation, the hearts in the two preserved groups were transplanted and studied as isolated hearts. Total ex vivo time for the preserved hearts was 24.5 +/- 0.2 hours. During preservation, glucose was not consumed by hearts in either preserved group. Pyruvate was used by the hearts to which it was provided (22.9 +/- 2.7 mumol pyruvate x hour-1 x gm dry weight-1). Performance of transplanted surgical control hearts was not significantly different from that of fresh controls. Function of the pyruvate-preserved hearts was similar to that of the fresh and surgical controls except that left ventricular peak systolic pressure and peak rate of pressure development were lower and that left ventricular end-diastolic pressure was higher for the pyruvate-preserved hearts. The hearts preserved without pyruvate had significantly lower compliance and function compared to the other three groups, which was evident in all indexes of contractility and output. We conclude that hearts preserved with pyruvate-containing crystalloid medium had better postpreservation, posttransplantation function than hearts preserved without pyruvate, although there was slight loss of compliance and decreased contractile function in the pyruvate-preserved hearts compared to controls.

Fluorochemical emulsion APE-LM substantially improves cardiac preservation.

We determined the efficacy of a novel fluorochemical emulsion for long-term hypothermic preservation of hearts. Rat hearts were preserved for 12 h at 12 degrees C with use of continuous low-pressure coronary perfusion with one of three oxygenated media (n = 6 hearts/groups): an "extracellular" crystalloid solution; APE-LM, a novel fluorochemical emulsion of perfluoroperhydrophenanthrene in egg yolk phospholipid; and FC-43, the Fluosol-43 (Oxypherol) fluorochemical emulsion of perfluorotributylamine in Pluronic F68. The emulsion media contained the same components as the crystalloid medium. All three media contained 0.5% albumin. An isolated working heart perfusion system was used to quantify the function of preserved hearts and controls (fresh hearts, n = 6). The APE-LM-preserved hearts were not significantly different from control hearts in contractile function, output, and energetics during a 4-h 37 degrees C reperfusion period. The control and APE-LM-preserved hearts had significantly better performance than crystalloid- and FC-43-preserved hearts. All preserved hearts gained fluid during preservation. The edema of APE-LM-preserved hearts, but not that of the other two preserved groups, was reversed during 37 degrees C reperfusion. These data provide the first evidence that a unique fluorochemical emulsion improves long-term preservation of cardiac tissue and produces significantly better recovery of cardiac function after preservation. This salutary effect was specifically associated with APE-LM emulsion and may result from its high O2 capacity, its biologically compatible emulsifier, and its superior physical properties, which include very small emulsion particle size (0.1-0.15 micron), low viscosity, and minimal toxicity.

Differences in high-energy phosphate catabolism between the rat and the dog in a heart preservation model.

Concentrations of ATP and creatine phosphate were measured in rat and dog hearts preserved either by cold storage (procedure A) or by continuous hypothermic perfusion (procedure B). In procedure A (3 dogs, 4 rats) the hearts were normothermically excised without cardioplegia and were stored in 0.9% NaCl at 0.5 degrees C; in procedure B (6 dogs, 21 rats) hypothermic cardioplegic arrest was performed, and then the hearts were retrogradely perfused through the aorta for 24 hours with use of an oxygenated Bretschneider cardioplegic solution at 2 degrees to 4 degrees C. Whole rat hearts were frozen using Wollenberger clamps at desired times during the preservation period; transmural needle biopsy specimens were sampled from dog hearts. In control nonpreserved hearts, the ATP and creatine phosphate were as follows (mean +/- SD): 26.7 +/- 4.1 and 27.1 +/- 10.3 mumol/gm dry weight, respectively (dog hearts), and 23.1 +/- 2.1 and 34.2 +/- 12.1 mumol/gm dry weight, respectively (rat hearts). With procedure A, ATP decreased by 36% in dog hearts and by 64% in rat hearts during the first hour of storage. By 24 hours, only 6% of the ATP remained in the dog hearts and 1% in the rat hearts. Creatine phosphate decreased by 85% (dog hearts) and by 93% (rat hearts) during the first hour of storage. The ATP and creatine phosphate values observed in rat hearts after 1 hour of procedure A preservation were significantly lower than in dog hearts (p less than 0.05). With procedure B, cardioplegic arrest by itself did not alter high-energy phosphate concentrations in dog or rat hearts.(ABSTRACT TRUNCATED AT 250 WORDS)

Attenuation of vasopressin-mediated coronary constriction and myocardial depression in the hypoxic heart.

To investigate the ability of arginine vasopressin (AVP) to compete with metabolic vasodilatory factors in the coronary circulation, we examined the coronary vascular and myocardial effects of AVP in isolated working rat hearts during normoxic and hypoxic perfusion. In normoxic hearts, AVP treatment (777 +/- 67 pg/ml) reduced coronary flow by 38.4 +/- 2.6%. Myocardial function was also significantly decreased by AVP whereas efficiency significantly increased. In contrast, the same dose of AVP administered to hypoxic hearts resulted in substantially smaller effects on coronary flow (-11.5 +/- 2.8%), myocardial function, and efficiency. In hearts treated first with AVP and then with hypoxia, the greater degree of coronary vasodilation compared with that observed in hearts treated with hypoxia alone also indicated an antagonizing effect of hypoxia on AVP-mediated coronary constriction. It was also noted that the hypoxia treatment alone resulted in reductions of O2 supply and consumption identical to those produced by AVP treatment during normoxia. However, hypoxia was associated with a significantly greater effect on myocardial function and, in contrast to the effect of AVP, a marked reduction in efficiency. The rate of lactate release was greater during hypoxia alone (2.07 +/- 0.08 mumol/min) than with AVP treatment during normoxia (0.76 +/- 0.05 mumol/min). These results indicate that the effect of AVP on the coronary vessels, as well as its effect on the myocardium, is significantly attenuated during hypoxia. In addition, AVP-constricted vessels appear to retain considerable vasodilatory reserve despite evidence of ischemic conditions. Thus, although the effects of AVP resemble ischemia, the increased efficiency and the relatively small effect of AVP on contractile function, as well as the preserved vasodilatory reserve, suggest otherwise. A physiological explanation for these observations is proposed wherein the constricting effects of AVP modulate the effects of autoregulatory factors such that blood flow requirements are minimized while allowing preservation of adequate blood flow for vital tissue function.

Tolerance to ethanol and cross-tolerance to pentobarbital by isolated hearts from chronic alcoholic rats.

The effects of ethanol and pentobarbital on the function of isolated working hearts from control and ethanol-fed male Long-Evans rats were studied. Hearts from ethanol-fed animals (38% of calories; 10-12 months) exhibited functional tolerance to the cardiodepressive effects of 18-71 mM ethanol in the perfusing medium. Left ventricular systolic pressure, peak relaxation rate, isovolumic pressure indexes, peak aortic flow rate, cardiac output, and stroke work of the control hearts were depressed to a greater extent than were those indexes of the alcoholic rat hearts during exposure to ethanol in vitro. Differences in the functional responses of controls and alcoholics were not the result of differences in energetics; myocardial O2 consumption, O2 supply-to-utilization ratio, and external work efficiency changed similarly in both groups of hearts during perfusion with ethanol. Cross-tolerance of the alcoholic rat hearts to the in vitro cardiodepressive effects of pentobarbital was also apparent. Peak rate of left ventricular pressure development, peak aortic flow rate, isovolumic pressure indexes, and cardiac output of the control rat hearts were significantly lower than those indexes of the alcoholic rat hearts during perfusion with 0.5 mM pentobarbital. In addition, four of the seven control hearts could not be paced during pentobarbital perfusion using 3V electrical pulses; the pacing voltage had to be raised and right ventricular pacing used to maintain stable function of those hearts at a 321 beats/min pacing rate. Myocardial O2 consumption and O2 supply-to-utilization ratios of control and alcoholic rat hearts were similar, but external work efficiency was slightly higher in the alcoholics than in controls during pentobarbital perfusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Albumin improves stability and longevity of perfluorochemical-perfused hearts.

We determined the effect of protein and nonprotein oncotic agents on physiological function and substructural integrity of perfluorochemical emulsion-perfused isolated working rabbit hearts. We used four media that contained Fluosol-43 (FL) emulsion and either 3.4% hydroxyethylstarch (HES), 2.9% albumin, 0.8% HES, or neither HES nor albumin (n = 5 hearts/group). All four groups of hearts had stable function for the first 5.5 h of perfusion; the FL plus albumin hearts continued to exhibit stability in most indexes of function until 9.5 h. The FL plus albumin hearts had a longer total period of ejecting function (12.5 +/- 0.5 h) compared with the other groups (mean longevities = 7.4-8.4 h). Functional stability and longevity correlated with maintenance of coronary flow and coronary vascular resistance. The rates of excess fluid accumulation and creatine kinase leakage were lower in the FL plus albumin hearts than in the other groups. We conclude that: 1) albumin maintained function, coronary flow, and myocardial cell integrity of FL-perfused hearts better than did HES; 2) albumin may exert its effect by preserving capillary permeability, thereby reducing the rate of interstitial fluid accumulation and preventing edema-induced vascular compression; and 3) HES had no effect on cardiac function or integrity and was ineffective in preventing interstitial fluid accumulation when it was used in FL-perfused isolated hearts in the absence of protein.

Direct effects of hydralazine on cardiac contractile function, haemodynamics, and myocardial energetics in isolated myocardium.

The direct cardiac effects of hydralazine were studied in isolated working rat heart, isolated cat right ventricular papillary muscle, and isolated rabbit right atrium. The haemodynamics, myocardial energetics, and contractility of isolated hearts were measured at hydralazine concentrations of 0.01, 0.1, 0.5, 1.0, 10 and 100 mumol.litre-1. Coronary flow was significantly increased (greater than or equal to 21%, p less than 0.01) in paced (325 beats.min-1) rat hearts at greater than or equal to 0.5 mumol.litre-1 hydralazine and in spontaneously beating hearts (greater than or equal to 37%; p less than 0.05) at greater than or equal to 1.0 mumol.litre-1 hydralazine. The increases in coronary flow occurred without significant increases in heart rate, contractility (dP/dtmax), or coronary perfusion pressure. Myocardial oxygen consumption was not significantly changed at any hydralazine concentration in spontaneously beating hearts and was unaltered in paced hearts except for a small significant increase (9.8%) at 10 mumol.litre-1. A negative inotropic effect was apparent at 100 mumol.litre-1 hydralazine as indicated by a significant reduction of dP/dtmax (paced and non-paced hearts), peak aortic flow rate (non-paced), and maximum left ventricular pressure (paced). In isolated cat papillary muscles and rabbit right atria, cumulative hydralazine log dose-response curves (0.1-1000 mumol.litre-1) were obtained. A positive inotropic effect that could be abolished by beta adrenergic blockade was produced in papillary muscles only at concentrations greater than or equal to 100 mumol.litre-1.(ABSTRACT TRUNCATED AT 250 WORDS)

A comparison of two anesthetic techniques for the study of rat skeletal muscle high-energy phosphates in vivo using 31P-NMR.

31P-Nuclear magnetic resonance (NMR) spectroscopy was used to evaluate in vivo rat quadriceps ATP, phosphocreatine (PCr), inorganic phosphate (Pi) and tissue pH during anesthesia with ketamine/xylazine (K/X) or isoflurane (IS). A surface coil was used to receive signals from the quadriceps muscle of rats positioned in a wide-bore horizontal magnet. The PCr/beta-ATP ratios determined from the NMR spectra were 4.34 +/- 0.19 (K/X) and 4.40 +/- 0.28 (IS). Tissue pH was 7.09 +/- 0.05 (K/X) and 7.13 +/- 0.07 (IS). Metabolic stability of quadriceps PCr and ATP was demonstrated during both K/X and IS anesthesia, but the K/X-anesthetized animals had longer sleep time, lower food consumption, and lower body weight post-anesthesia than the IS-anesthetized animals. The PCr/beta-ATP ratio in quadriceps of repetitively IS-anesthetized rats did not fluctuate diurnally. In addition, the animals recovered rapidly and continued to gain weight following the multiple brief IS anesthetic procedures. These data indicate that serial in vivo investigations of high-energy-phosphate metabolism in small animals can be accomplished using 31P-NMR spectroscopy and IS anesthesia, which has several advantages over K/X anesthesia for these types of studies.

Chronic alcohol treatment results in disturbed vitamin D metabolism and skeletal abnormalities in rats.

The effect of chronic alcohol consumption on the skeleton was investigated in rats. The treated group received ethanol administered as 38% of caloric intake in a liquid diet (Sustacal) for 10 months. The control rats were pair weighted to the ethanol-treated animals throughout the study; the growth curves of the two groups were the same. The controls were given the same liquid diet except that dextrin:maltose (3:1) was substituted isocalorically for ethanol. Ethanol-treated rats did not differ from the pair-weighted controls in mean serum calcium, phosphorous, or creatinine. In contrast, serum magnesium was reduced (p less than 0.02) in alcohol-treated rats. Ethanol treatment also resulted in changes in the serum concentrations of vitamin D metabolites; serum 25-hydroxyvitamin D3 was increased (p less than 0.001), while serum 1,25-dihydroxyvitamin D3 was decreased (p less than 0.01). Tibial length was reduced in ethanol-treated rats (p less than 0.05) but there was no change in femoral length. Medullary area was increased in tibial diaphyses from alcohol-treated rats compared to weight matched control animals (p less than 0.01), indicating a net increase in resorption. The cross-sectional area of the tibial diaphysis of ethanol-treated rats was the same as the matched controls. Trabecular bone was decreased in the tibial metaphysis of ethanol-treated rats compared to the matched controls (p less than 0.05) indicating a net loss of trabecular bone. Ethanol treatment did not have an effect on the organic weight of the femur but the ash weight was reduced (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

The development of alcohol-induced cardiac dysfunction in the rat.

Hemodynamic, contractile and energetic functions of isolated working hearts from ethanol-fed and control rats were studied to determine the time course of cardiac dysfunction development during long-term consumption of alcohol. Hearts from fasted, 24 hr withdrawn rats were studied after 2, 4, 7 and 11 months consuming ethanol as 38% of daily calories in a nutritionally-adequate liquid diet. After 2 months, the right ventricle of the alcoholic rat hearts was significantly enlarged; left ventricular weight was not significantly different from control and there was little evidence that left ventricular function was compromised. After 4 months and 7 months, there was biventricular cardiomegaly and evidence of reduced left ventricular function in the alcoholics, although altered sensitivity to the positive inotropic agent, dobutamine, was not evident in those hearts. After 11 months, cardiac output, stroke work, and peak power of the alcoholic rat hearts were significantly depressed, the responsiveness of the left ventricle to dobutamine was diminished, and both ventricles were enlarged compared to controls. Cardiac function during early withdrawal was studied in rats that had consumed alcohol for 14-16 months. Hearts from non-fasted rats at 0 hr of withdrawal exhibited diminished responsiveness to dobutamine compared to controls; at 24 hr and 72 hr of withdrawal no differences between alcoholic and control rat heart dobutamine responsiveness were observed. The data indicated that: (a) cardiomegaly, beginning with right ventricular enlargement, was an early indicator of alcohol's cardiac effects in rats; (b) left ventricular enlargement of the alcoholic rat hearts was associated with basal left ventricular dysfunction; and (c) evidence of cardiac subsensitivity to dobutamine, which is characteristic of this alcoholic rat model, depended on the nutritional status and withdrawal state of the rat at the time that the heart was excised for study.

Alcoholic cardiomyopathy in rats: inotropic responses to phenylephrine, glucagon, ouabain, and dobutamine.

The inotropic responses of chronic alcoholic and control rat hearts to phenylephrine, glucagon, ouabain, and dobutamine were studied to determine if the reported beta-adrenergic subsensitivity of alcoholic rat hearts was a specific defect. Male Long-Evans rats were maintained on nutritionally-complete liquid diets for 10 to 12 months; alcoholic rats received 38% of their calories from ethanol. Dry heart weight/body weight ratios indicated an average 15% hypertrophy of the alcoholic rat hearts. The function of isolated working hearts from these animals was studied at a constant heart rate and afterload. Ventricular function curves indicated significantly lower basal function of alcoholic rat hearts, as evident from their lower peak left ventricular relaxation rate, lower isovolumic relaxation rate, and lower peak power compared to controls. The alcoholic rat hearts had significantly lower inotropic (stroke work and peak power) responses to phenylephrine, glucagon, and dobutamine compared to controls, whereas the response of the alcoholics to ouabain was not significantly different from that of controls. Oxygen supply-to-utilization ratios decreased similarly in alcoholics and controls during treatment with the inotropic agents, as a result of increases in myocardial oxygen consumption and effects on coronary flow that were similar in both groups of animals. Thus the differences in inotropic responses observed with the alcoholic rat hearts were not primarily the result of compromised oxygen supply. Rather, the decreased stroke work response of the alcoholic hearts which occurred despite an increase in oxygen consumption suggested that the alcoholic rat hearts did not utilize oxygen as efficiently as did control hearts to perform external work. This was reflected in the significant differences between alcoholics and controls in the response of calculated external work efficiency to phenylephrine, glucagon, and dobutamine. Thus, alcohol-induced cardiac hypertrophy was associated with depressed basal left ventricular contractile function and decreased responsiveness to alpha 1-adrenergic, beta 1-adrenergic, and glucagon stimulation, but the responsiveness to ouabain was not significantly affected. These characteristics are similar to those of hearts hypertrophied by other causes.

Prolonged support of working rabbit hearts using Fluosol-43 or erythrocyte media.

We compared the perfluorochemical emulsion Fluosol-43 and an erythrocyte-based solution as support media for ex vivo working rabbit hearts functioning with a physiological workload. Both groups of hearts (n = 5/group) exhibited stable function (left ventricular peak systolic pressure, peak rates of left ventricular pressure rise and relaxation, aortic flow, peak aortic flow rate, stroke work, and peak power) for the first 6 h of perfusion. Coronary flow, coronary venous O2 content, and O2 supply-to-demand ratio declined similarly in both groups during the first 6 h. Both groups of hearts preferentially utilized pyruvate to glucose. The Fluosol-43-perfused hearts had higher heart rate, left ventricular peak systolic pressure, peak rate of left ventricular pressure rise, aortic flow, coronary flow, and myocardial O2 consumption compared with the erythrocyte-perfused hearts. The Fluosol-43 hearts produced more lactate and released more creatine phosphokinase than did the erythrocyte-perfused hearts, but the rates were low and constant throughout perfusion, indicating that the hearts were not progressively ischemic. After the first 6 h, function of the Fluosol-43 hearts declined, resulting in their earlier failure compared with the erythrocyte-perfused hearts. The data indicate that Fluosol-43 had sufficient O2- carrying capacity to support stable function of a rabbit heart at a physiological workload for 6 h, and differences in function and ex vivo longevity of the two groups of hearts suggested that a component or contaminant of Fluosol-43 altered sarcolemmal function and/or that a component needed for membrane integrity was lacking in the Fluosol-43 perfusate.

Direct cardiac effects of vasopressin and their reversal by a vascular antagonist.

We studied the direct cardiac effects of arginine vasopressin (AVP) by use of an isolated working rat heart model perfused with Krebs-Henseleit medium. At a concentration of 878 +/- 15 pg/ml, AVP produced significant (P less than 0.05) decreases in coronary flow (-31 +/- 2%); myocardial O2 consumption (-12 +/- 2%); left ventricular peak systolic pressure (-5 +/- 1%); dP/dtmax (-7 +/- 1%); -dP/dtmax (-6 +/- 3%); peak aortic flow rate (-5 +/- 1%); stroke work (-3 +/- 1%); peak power (-8 +/- 1%); and total output (-3 +/- 1%). Aortic output increased significantly (+7 +/- 1%) as did arteriovenous O2 difference (+108 +/- 14 mmHg); left ventricular end-diastolic pressure (+0.4 +/- 0.1 mmHg); efficiency (+1.5 +/- 0.4%); and rate of lactate release (+1.27 +/- 0.21 nmol/ml perfusate/min). Dose-response relationships were studied at 9 +/- 1, 25 +/- 1, 75 +/- 3, 303 +/- 15, and 817 +/- 42 pg AVP/ml. Significant dose-dependent depression of coronary flow occurred at the three highest AVP concentrations; cardiac function was significantly depressed at the highest dose. The AVP analogue d(CH2)5[Tyr(Me)]AVP (20 ng/ml) completely reversed the cardiac effects attributed to AVP. The data indicate that AVP is a potent direct coronary constrictor that produces myocardial ischemia and decreased contractile function at AVP concentrations that are observed in some pathophysiologic states.(ABSTRACT TRUNCATED AT 250 WORDS)