BACKGROUND:Precise estimation of the cellular water content presupposes a correct definition of the water fraction in tissue extracellular space. Low molecular weight markers (LMM), such as sulphate ion and sucrose, are widely used to define extracellular space size despite indications that they penetrate the cell. In contrast, inulin, with molecular weight of about 5000, is commonly regarded as a cell impermeable extracellular marker.OBJECTIVES:To compare LMM with inulin as markers in determining extracellular space size.ANIMALS AND METHODS:The size of extracellular space in guinea pig hearts perfused with crystalloid solution (hydrated hearts) was determined morphometrically and by mathematical model analysis of washout kinetics of LMM ((35)SO(4), (14)C-sucrose) or (3)H-inulin.RESULTS:Morphometrically, the sizes of vascular and interstitial spaces in the hydrated hearts were estimated to be 102+/-8 mL/kg wet mass (wm) and 452+/-17 mL/kg wm, respectively. Comparable data were obtained from model simulation of tracer washout: 67 mL/kg wm for vascular space and 439 to 462 mL/kg wm for interstitial space. Tracer penetration into cellular water, as shown by model analysis, was 28% for LMM and, reported here for the first time, 18% for inulin. The observed edema was probably due entirely to fluid accumulation in the interstitial space.CONCLUSION:Intracellular penetration of LMM must be taken into account, especially in modern nuclear magnetic resonance spectroscopic methods of cellular water monitoring in isolated perfused hearts.

Effects of adaptation to hypoxia on the contractility of isolated rat hearts and on their levels of ATP and inorganic phosphate after total ischemia were evaluated. This adaptation failed to render the cardiac energy-supplying system more resistant to postischemic reperfusion and thus did not accelerate the restoration of cardiac contractility after ischemia. The results of adaptation to hypoxia were then compared with those of adaptation to stress, which had been shown to bring about a marked increase in cardiac resistance to postischemic reperfusion. It is concluded that the profound differences noted between the cardioprotective effects of these two forms of adaptation are due to a much greater accumulation of stabilizing proteins from the HSP70 family during adaptation to stress.

Effects of 2,3-butanedione monoxime (BDM), an ATPase inhibitor, on ischemia-reperfusion myocardial injury were examined in isolated working rat hearts perfused in vitro. Following cardiac arrest induced by cardioplegic solution, global ischemia was produced for 30 min. In untreated hearts, reperfusion for 45 min resulted in an incomplete recovery of cardiac pump function. When BDM was added to the cardioplegic solution up to 20 mM, the recovery of cardiac function was significantly improved average by 19%. This BDM effect can, presumably, reduce ATP losses during ischemia and like that improve recovery of cardiac function during reperfusion.

Isolated Langendorff-perfused guinea pig hearts were arrested with a cardioplegic solution containing 10 mM phosphocreatine + 15 mM glutamate (PG group) or not containing them (control group). Total normothermic ischemia lasted 45 min followed by 30 min reperfusion. Mitochondrial respiration in the absence and presence of different concentrations of ADP and creatine was studied in biopsy samples after saponin treatment. The samples were taken before and after ischemia as well as after the reperfusion period. A slightly better relative recovery of developed pressure (RRDP) in PG group was associated with higher mitochondrial acceptor control ratio after reperfusion. When results in both groups were taken together, marked negative correlations between the preischemic mitochondrial indices (particularly, those related to creatine kinase activity) and RRDP were revealed. Relative changes in these indices after ischemia demonstrated tight positive correlations with RRDP. Thus, the hearts having higher functional activity of mitochondrial creatine kinase are more sensitive to ischemia, other conditions being equal.

Isolated perfused guinea pig hearts were arrested by a high K+ cardioplegic solution containing (PG group) or lacking (control group) 10 mM phosphocreatine +15 mM glutamate. Total normothermic ischemia lasted 45 min followed by 30 min reperfusion. Mitochondrial respiration in the absence and presence of different concentrations of ADP and creatine was studied in biopsy samples (6–8 mg) after saponin treatment. The samples were taken before and after ischemia, as well as after the reperfusion period. A slightly better relative recovery of developed pressure (RRDP) in PG group was associated with higher mitochondrial acceptor control ratio after reperfusion (5.74±0.32 vs. 4.54±0.21 in PG and control groups, resp., p<0.01). When the results obtained in both groups were treated together, tight correlations between the pre- or postischemic mitochondrial state and RRDP were revealed. Higher values of RRDP were found for the hearts with lower preischemic values of (low ADP+creatine)-stimulation of mitochondrial respiration (r=−0.57, p<0.01). Relative changes in this mitochondrial parameter during ischemic period were in a good correlation with the RRDP (r=0.82, p<0.001). The data suggest that the study of the mitochondrial function in myocardial biopsy samples before ischemia and reperfusion could provide a useful information for the prognosis of cardiac function recovery.

The rats receiving a cumulative dose of adriamycin, 15-20 mg/kg exhibited delayed weight gain, ascites, and increased relative liver weight. The animals' isolated hearts perfused at a constant rate in the Krebs' solution in a retrograde fashion showed lowered perfusion pressure, indicating reduced coronary vascular tone. At the constant heart rate, the hearts from the adriamycin-treated animals consumed oxygen less at equally increased pressure. In total ischemia, higher cellular lactate and K+ yield occurred than in controls, the severity of acidosis being nearly similar. Prior to ischemia, the pressure produced by the hearts from adriamycin-treated animals was less but, following reperfusion, it more rapidly became normal than that in controls. The results suggest a compensatory increase in myocardial glycolysis during chronic administration of adriamycin.

The response of rat and guinea-pig hearts to ischemia and reperfusion has been studied in identical conditions. Total 15-min ischemia of isolated rat hearts at 36 degrees C induced an almost 3-fold rise in isovolumic left ventricular diastolic pressure as well as a fall in the developed pressure and heart rate. Guinea-pig hearts, in the same conditions, exhibited a more steep fall in heart rate, with no rise in diastolic pressure. With constant heart rate produced by electrical stimulation at 4 Hz, the difference between two groups remained unchanged, while a more rapid fall in developed pressure in guinea-pig hearts coincided with a more profound fall in extracellular pH and almost a 2-fold rise in extracellular K+ activity. Rapid elimination of K+ and H+ at the early stages of reperfusion was followed by fibrillation in the majority of guinea-pig hearts, while no fibrillation was observed in rat hearts.