Failure to maintain a low ADP concentration impairs diastolic function in hypertrophied rat hearts.

BACKGROUND: Mechanisms in addition to diastolic calcium overload may contribute to diastolic dysfunction in hypertrophied hearts. In this study, we tested the hypothesis that failure to maintain a low ADP concentration in hypertrophied hearts contributes to diastolic dysfunction by inhibiting the rate of cross-bridge cycling. METHODS AND RESULTS: By perfusing isolated rat hearts with pyruvate and 2-deoxyglucose (2DG), we were able to perturb [ADP] with minimal changes in [ATP] and [inorganic phosphate] or the contribution of glycolytic ATP to ATP synthesis. The effects of 2DG were compared in aortic-banded (LVH, n=5) and sham-operated (control, n=5) rat hearts. 31P NMR spectroscopy was used to measure the concentrations of phosphorus-containing compounds. We found a threefold increase of left ventricular end-diastolic pressure (LVEDP) in LVH during 2DG perfusion, and this increase was concomitant with a threefold increase in intracellular free [ADP]. The [ADP] in the control hearts was maintained <40 micromol/L, and no change in LVEDP was observed. A linear relationship between increases in [ADP] and LVEDP was found (r2=.66, P=.001). Furthermore, the capacity of the creatine kinase reaction, a major mechanism for maintaining a low [ADP], was decreased in LVH (P=.0001). CONCLUSIONS: Increased [ADP] contributes to diastolic dysfunction in LVH, possibly due to slowed cross-bridge cycling. Decreased capacity of the creatine kinase reaction to rephosphorylate ADP is a likely contributing mechanism to the failure to maintain a low [ADP] in LVH.

NMR of laser-polarized 129Xe in blood foam.

Laser-polarized 129Xe dissolved in a foam preparation of fresh human blood was investigated. The NMR signal of 129Xe dissolved in blood was enhanced by creating a foam in which the dissolved 129Xe exchanged with a large reservoir of gaseous laser-polarized 129Xe. The dissolved 129Xe T1 in this system was found to be significantly shorter in oxygenated blood than in deoxygenated blood. The T1 of 129Xe dissolved in oxygenated blood foam was found to be approximately 21 (+/-5) s, and in deoxygenated blood foam to be greater than 40 s. To understand the oxygenation trend, T1 measurements were also made on plasma and hemoglobin foam preparations. The measurement technique using a foam gas-liquid exchange interface may also be useful for studying foam coarsening and other liquid physical properties.

Creatine kinase system in failing and nonfailing human myocardium.

BACKGROUND: The creatine kinase (CK) reaction is important for rapid resynthesis of ATP when the heart increases its work. Studies defining the CK system in human failing and nonfailing myocardium are limited and in conflict. To resolve this conflict, we measured the activities of CK and its isoenzymes and the contents of creatine and CK-B in homogenates of human myocardium. METHODS AND RESULTS: Myocardium was sampled from 23 subjects who underwent heart transplant, 36 subjects maintained in an intensive care unit before heart harvesting, 13 accident victims, and 2 patients undergoing heart surgery. Since the characteristics of myocardium of potential organ donors differed from those of myocardium of accident victims, data are presented for three groups: failing, donor, and control. CK activity was 7.7 +/- 1.9 and 6.0 +/- 1.4 IU/mg protein in left (LV) and right (RV) ventricles of failing, 9.4 +/- 2.5 and 10.7 +/- 2 IU/mg protein in LV and RV of donor, and 11.6 +/- 2.4 IU/mg protein in LV of control hearts. CK-MM and the mitochondrial isoenzyme activities were lower in failing and donor LV, and CK-MB activity and CK-B content were higher in failing and donor hearts. Creatine contents were 64 +/- 25 and 56 +/- 18.6 nmol/mg protein in LV and RV of failing, 96 +/- 30 and 110 +/- 24 nmol/mg protein in LV and RV of donor, and 131 +/- 28 nmol/mg protein in LV of control hearts. CONCLUSIONS: In failing and nonfailing donor human myocardium, there is a combined decrease of CK activity and creatine that may impair the ability to deliver ATP to energy-consuming systems.

Determinants of tissue delivery for 129Xe magnetic resonance in humans.

Magnetic resonance imaging using laser-polarized 129Xe is a new technique first demonstrated by Albert et. al. (Nature 370, 1994) who obtained a 129Xe image of an excised mouse lung. This paper describes the factors influencing the accumulation of inhaled, polarized 129Xe in human tissue. The resulting model predicts the 129Xe magnetization in different tissues as a function of the time from the start of inhalation, the tissue perfusion rate and partition coefficient for xenon, and the relevant T1 decay times. The relaxation times of 129Xe in biological tissues are not yet known precisely. Substitution of estimated values for these parameters results in an expected signal-to-noise ratio (SNR) from polarized 129Xe MR in the brain of approximately 2% of the equivalent SNR from proton MR.

Decreased energy reserve in an animal model of dilated cardiomyopathy. Relationship to contractile performance.

An animal model was used to test the hypothesis that in heart failure the decrease in the ability to resynthesize ATP through the creatine kinase (CK) reaction (which we call energy reserve) contributes to the inability of the heart to maintain its normal function and contractile reserve. One-week-old turkey poults were fed furazolidone for 14 days to induce dilated cardiomyopathy. Isolated Langendorff-perfused hearts from these myopathic animals showed a 73% decrease in baseline isovolumic contractile performance. Neither increasing [Ca2+]o nor electrical pacing rate increased isovolumic contractile performance. Measured by 31P nuclear magnetic resonance magnetization transfer and chemical assay, ATP concentration was decreased by 23%, phosphocreatine concentration by 42%, CK enzyme activity by 34%, and the pseudo first-order rate constant for the CK reaction by 50%. Measured CK reaction velocity decreased by 71%. The reduced ability to increase cardiac performance in response to increasing [Ca2+]o in hearts with lower CK reaction velocity was reproduced in part by feeding a separate group of turkey poults beta-guanidino-propionic acid to specifically reduce CK reaction velocity by decreasing guanidino substrate concentration. These hearts had normal baseline performance but blunted contractile reserve. These observations provide further support for the hypothesis that a decrease in energy reserve via the CK system contributes to reduced cardiac function in the failing heart.

Depletion of energy reserve via the creatine kinase reaction during the evolution of heart failure in cardiomyopathic hamsters.

To study the contribution of myocardial energy reserve to the deterioration of cardiac function during the development of heart failure, we defined energy reserve via the creatine kinase (CK) reaction and the isovolumic contractile performance in hearts of cardiomyopathic hamsters at the ages of 1.5, 4, 17, 30 and 43 weeks and in age-matched normal hamsters. Energy reserve via the CK reaction was estimated by the product of total CK activity and the content of total creatine in the heart. Isovolumic contractile performance was measured as rate pressure product (RPP, 10(3) mmHg/min) in isolated hearts. Contractile reserve was assessed as the increase of RPP elicited by high calcium stimulation. Compared to the controls, decreases in total CK activity and content of total creatine were observed in hearts of 17-, 30- and 43-week-old cardiomyopathic hamsters. These changes were not observed in the skeletal muscle. Although the decrease of baseline RPP first occurred at the age of 30 weeks (11.5 +/- 0.9 v 20.5 +/- 0.8, P < 0.05), the contractile reserve was already reduced at the age of 17 weeks (9.9 +/- 1.3 v 23.6 +/- 1.9, P < 0.05). A linear relationship was found between the energy reserve via creatine kinase reaction and the contractile reserve of the heart (r2 = 0.85). Furthermore, concomitant decreases in the CK reaction velocity and the contractile reserve were observed in cardiomyopathic hearts, suggesting that depletion of energy reserve may contribute to the development of heart failure.

Enalapril treatment increases cardiac performance and energy reserve via the creatine kinase reaction in myocardium of Syrian myopathic hamsters with advanced heart failure.

BACKGROUND: Converting enzyme inhibitor treatment of congestive heart failure slows progression to failure and reduces mortality rate. It is known whether these benefits are due solely to improved hemodynamics or also to improved myocyte energetics. This study examines the effect of enalapril treatment on both isovolumic contractile performance and its biochemical correlate, flux through the creatine kinase (CK) system, in an animal model of severely failing myocardium. METHODS AND RESULTS: Seven-month-old Syrian cardiomyopathic (TO-2 strain) and normal golden Syrian (FIB strain) hamsters were each randomly assigned to one of three groups supplied daily with either no, low (25 mg/kg body wt), or high (100 mg/kg body wt) doses of enalapril for 12 to 14 weeks. At 10 months of age, all substrates and products and flux through the CK reaction were measured in isolated perfused hearts by 31P magnetization transfer and chemical assay. Compared with normal hamsters, the myopathic hamsters exhibited significantly lower body weights and higher biventricular heart weights, which were partially reversed by drug treatment. The Langendorff-perfused hearts showed decreased isovolumic contractile performance with identical load conditions. This was partially reversed by drug treatment. In the failing hearts, the following substrate and product concentrations and enzyme activities were decreased compared with nonfailing hearts but were unchanged by drug treatment: ATP (-28%), phosphocreatine (-48%), free creatine (-64%), ADP (-51%), and CK (-34%, primarily MM isoenzyme). Flux through the CK reaction for the untreated cardiomyopathic hamster hearts was decreased by 67%, and this decrease was almost completely reversed by enalapril treatment. The increased CK flux is due to an increase in the rate constant for the reaction, since substrate concentrations are unchanged, and is not predicted by the rate equation. In enalapril-treated failing hearts, phosphoryl transfer via the CK reaction increased with contractile performance. This was not observed in the nonfailing hearts, in which energy reserve is adequate to support changes in contractile performance. CONCLUSIONS: Decreased flux through CK reaction leads to decreased capacity for ATP synthesis and may contribute to decreased contractile performance in cardiomyopathic hamster hearts. Enalapril treatment results in increased phosphoryl transfer through the CK reaction in failing myocardium, and this increase is coupled to improved cardiac performance. Decreased CK flux in failing hearts is due to a combination of decreased Vmax and lower guanidino pool; this mechanism fails to explain changes in CK flux in enalapril-treated failing hearts.

Phosphocreatine T1 measurements with and without exchange in the heart.

The intrinsic phosphocreatine (PCr) T1 values measured by time-dependent magnetization transfer in isolated perfused rat, hamster, and turkey hearts were indistinguishable. The value of 3.5 +/- 0.3 s for the rat heart is similar to values measured by other magnetization transfer methods. Irreversibly inhibiting the phosphoryl exchange between PCr and ATP in the rat heart using iodoacetamide changed the apparent T1 values of the two exchanging species when measured by inversion recovery: The apparent T1 of PCr increased from 1.92 +/- 0.06 s to 3.55 +/- 0.06 s, in excellent agreement with the intrinsic T1 measured by magnetization transfer. The apparent T1 of [gamma-P]ATP decreased from 0.92 +/- 0.07 s to 0.44 +/- 0.03 s. The value for the T1 of [gamma-P]ATP in hearts with inhibited phosphoryl exchange was similar to T1 values for [alpha-P]ATP and [beta-P]ATP, which remained unchanged. This illustrates that apparent T1 values for PCr and [gamma-P]ATP measured by inversion recovery in the presence of exchange are average T1 values in between the intrinsic values. The large differences between the intrinsic T1 measured by magnetization transfer and the T1 measured by inversion recovery makes the use of the appropriate value in different applications quantitatively important.

Changes in ventricular creatine-kinase with progression and regression of cardiac hypertrophy in hypertensive rats.

We studied the distribution of four isoenzymes of creatine-kinase (MM, MB, BB and the mitochondrial) in 13 renovascular hypertensive rats and five age-matched controls. After 6 weeks of hypertension, seven rats were treated with captopril (2 mg/kg daily) for 4 weeks and the other six were left untreated for 4 weeks. After the rats were killed an adaptive increase in MB and BB was found at the expense of MM in the hypertensive rats compared with the controls. The captopril-treated rats displayed persistently higher levels of both MB and BB than the controls. Therefore, the regression in cardiac hypertrophy that is achieved by captopril is accompanied by persistence of the isoenzymic pattern that leads to better use of energy-rich phosphates.

Ventricular myosin and creatine-kinase isoenzymes in hypertensive rats treated with captopril.

In the myocardium, myosin and creatine kinase isoforms possess different capacities for using O2 and energy-rich phosphates. We studied electrophoretically the distribution of these isoforms in 19 hypertensive rats (two-kidney, one clip model of hypertension) and in age-matched controls. After 6 weeks of hypertension, seven rats were treated with captopril (2 mg/kg daily) for 4 weeks, six were left hypertensive for another 4 weeks, and the remaining rats were killed under ether anesthesia. In the latter, ventricular mass was significantly higher than in controls; V3 isomyosin was 32.3 +/- 6.8% versus 0%, and both creatine kinase-MB and -BB were increased at the expense of creatine kinase-MM (creatine kinase-MB = 29 +/- 2.8% vs. 14.7 +/- 1.8%, p less than 0.001; creatine kinase-BB = 3.1 +/- 0.6% vs. 1.7 +/- 0.8%, p less than 0.001). After 10 weeks of hypertension, ventricular mass, V3 isomyosin, and both creatine kinase-MB and -BB isoforms were found to be persistently higher than in controls. At the same time, captopril-treated rats showed reduced but not normalized blood pressure levels, normalized ventricular mass, and prevalence of the V1 isomyosin (56.9 +/- 22% vs. 47.9 +/- 23.8% in normotensive controls, p = NS). However, higher levels of creatine kinase-MB and -BB were still found in these rats in comparison with the normotensive controls (creatine kinase-MB = 22.4 +/- 5.4% vs. 15.8 +/- 2.8%, p less than 0.025; creatine kinase-BB = 2.3 +/- 0.1% vs. 1.8 +/- 0.3%, p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

[Usefulness of the 12-lead standard electrocardiogram in assessing myocardial infarct size]. / Utilità dell'elettrocardiogramma standard a 12 derivazioni nella stima delle dimensioni dell'infarto miocardico.

The value of the electrocardiogram in assessing infarct size was studied using serial estimates of serum creatine phosphokinase (CPK) in serum and serial 12-lead electrocardiograms in patients admitted to a coronary care unit with acute myocardial infarction. Sum of the ST segment deviations from the isoelectric line (sigma ST12) and sum of the conventional scores of Q waves amplitude (sigma Q12) were obtained from each electrocardiogram, and then the time-course of these parameters was considered. The correlation between maximum sigma Q12 and CPK-peak in anterior infarcts, was highly significant (r = 0.733; P less than 0.001). Maximum sigma ST12, measured upon admission or immediately thereafter in patients hospitalized within 4 hours from the onset of chest pain, was found to correlate significantly with CPK-peak (r = 0.675, P less than 0.001 for the whole group; r = 0.758, P less than 0.01 for patients with inferior infarcts). Time-course of sigma ST12 and CPK showed 4 different patterns. Among them, type 1 ("rapid necrosis") showed the most significant correlations (maximum sigma ST12 within 4 hours from symptoms versus CPK-peak: r = 0.909, P less than 0.001; maximum sigma Q12 versus CPK-peak in anterior infarcts: r = 0.782, P less than 0,05; and maximum sigma ST12 within 4 hours from the onset of pain versus sigma Q12 in patients with anterior infarcts: r = 0.863, P less than 0,05). There was no correlation between sigma ST12 at any other time and CPK-peak: this observation is in accordance with the presence of a rapid decrease in the mean sigma ST12 after the first 3-4 hours from the beginning of symptoms. This study shows that the analysis of ST segment deviations and of Q waves development in the standard electrocardiogram provides useful information on the size of acute myocardial infarction as reflected by the peak value of serum CPK.

[ST-segment trend in patients with acute myocardial infarction treated with antithrombotic drugs (author's transl)]. / Decorso del tratto ST in pazienti con infarto miocardico acuto trattati con farmaci antitrombotici.

Fifty-five patients were hospitalized in a Coronary Care Unit within the first 24 hours after onset of symptoms of an acute myocardial infarction. The sum of positive and negative ST-segment deflections of their twelve leads electrocardiograms (epsilon ST12) showed a maximum within the 3rd hour from the onset of symptoms. Thereafter, in the first 24 hours, there was a marked reduction in epsilon ST12, with a steep and significant fall within the 7th hour from symptoms. In the following nine days of this study, the patients showed 4 different epsilon ST trends, but there was no significant correlation with CPK curves, or with the kind of therapy the patients underwent. An irregular trend of epsilon ST12 or a secondary late rise (after 36 hours from symptoms) suggest an unfavorable prognosis (1 death in the epsilon ST-3 group, and 2 deaths + 1 ventricular fibrillation in the epsilon ST-4 group). No significant difference results between the patients treated with high doses Heparin plus Acetyl-Salicylic-Acid (A.S.A.), and the patients treated with A.S.A alone, though the first treatment seems to reduce the values of epsilon ST12 more rapidly; perhaps this behaviour is due to the use of antiplatelet drug A.S.A in both groups, and to the relatively small number of patients.