Alterations of myocardial dynamic stiffness implicating abnormal crossbridge function in human mitral regurgitation heart failure.

Mitral regurgitation (MR) causes ventricular dilation, a blunted myocardial force-frequency relation, and increased crossbridge force-time integral (FTI). The mechanism of FTI increase was investigated using sinusoidal length perturbation analysis to compare crossbridge function in skinned left ventricular (LV) epicardial muscle strips from 5 MR and 5 nonfailing (NF) control hearts. Myocardial dynamic stiffness was modeled as 3 parallel viscoelastic processes. Two processes characterize intermediate crossbridge cycle transitions, B (work producing) and C (work absorbing) with Q(10)s of 4 to 5. No significant differences in moduli or kinetic constants of these processes were observed between MR and NF. The third process, A, characterizes a nonenzymatic (Q(10)=0.9) work-absorbing viscoelasticity, whose modulus increases sigmoidally with [Ca(2+)]. Effects of temperature, crossbridge inhibition, or variation in [MgATP] support associating the calcium-dependent portion of A with the structural "backbone" of the myosin crossbridge. Extension of the conventional sinusoidal length perturbation analysis allowed using the A modulus to index the lifetime of the prerigor, AMADP crossbridge. This index was 75% greater in MR than in NF (P=0.02), suggesting a mechanism for the previously observed increase in crossbridge FTI. Notably, the A-process modulus was inversely correlated (r(2)=0.84, P=0.03) with in vivo LV ejection fraction in MR patients. The longer prerigor dwell time in MR may be clinically relevant not only for its potential role as a compensatory mechanism (increased economy of tension maintenance and increased resistance to ventricular dilation) but also for a potentially deleterious effect (reduced elastance and ejection fraction).

Prevention by fructose-1,6-bisphosphate of cardiac oxidative damage induced in mice by subchronic doxorubicin treatment.

An experimental model of mild, subchronic doxorubicin cardiotoxicity in mice was investigated by monitoring changes of biochemical parameters related to cell response against oxidative stress in both liver and heart. A specific increase of the lactate dehydrogenase isoenzyme typical of the heart was observed for doxorubicin-treated mice. Lipid peroxidation, as evaluated by malondialdehyde determination, and catalase activity were greatly increased in heart and unaffected in liver. On the other hand, these changes can be considered as indicative of early heart damage induced by doxorubicin. Glutathione, glutathione peroxidase, and 6-phosphogluconate dehydrogenase values were not significantly altered by the treatment and glucose-6-phosphate dehydrogenase increased in both liver and heart. Administration of fructose-1,6-bisphosphate strongly reduced the increase of plasma lactate dehydrogenase, heart lipid peroxidation, and heart catalase while no effect on the diagnostically irrelevant increase of glucose-6-phosphate dehydrogenase was observed. The inhibitory effect on the onset of biochemical modification typical of early subchronic doxorubicin cardiotoxicity may be related to stimulation of ATP synthesis by fructose-1,6-bisphosphate and is therapeutically promising in view of the lack of toxicity of fructose-1,6-bisphosphate as a drug.

Reticuloendothelial system activated by fructose-1,6-diphosphate in mice.

Fructose-1,6-diphosphate (FDP) exerts a marked activity on reticuloendothelial system (RES) functions, increasing in mice the clearance of colloidal carbon. ATP depletion occurring during phagocytic activity is concentrated by FDP.

The inhomogeneity and appropriateness of the myocardial response to stress.

Myocardial hypertrophy, with high morbidity and mortality, is a natural outcome of hypertensive heart disease. The increase in myocardial mass is associated with a cellular and subcellular reorganization of the myocytes. The following study uses rapid myothermal techniques to assess the contribution of the major intracellular changes to the adaptive hypertrophic process in various heart models. Pressure overload and thyrotoxic hypertrophy were produced in the rabbit. In the rat, hypertrophy was produced by constricting the renal artery (Goldblatt hypertensive rat) or by using the spontaneously hypertensive rat strain. Atrophy was produced by administration of propylthiouracil in the drinking water. The V1/V3 myosin isoenzyme ratio was decreased in the pressure overload, Goldblatt, and propylthiouracil animals. This was associated with a decrease in total activity-related heat, initial heat, and tension-dependent heat per tension time integral. The tension-independent heat was decreased in the pressure overload, while the time to peak tension was increased. The economy of the metabolic recovery process was unchanged in the pressure overload and Goldblatt preparations. In the propylthiouracil preparation the recovery processes became uneconomical. The spontaneously hypertensive rat exhibited mild cardiac hypertrophy but in all other respects the heart was unchanged from the normal animals. The thyrotoxic hearts had a high V1/V3 myosin isoenzyme ratio, which was associated with a high total activity-related heat, initial heat, and tension-dependent heat per tension time integral. The tension-independent heat was reduced in the thyrotoxic preparations. The appropriateness of each of the intracellular changes is evaluated in terms of the demands made on the heart.

Dynamic calcium requirements for activation of rabbit papillary muscle calculated from tension-independent heat.

The heat generated by right ventricular papillary muscles of rabbits was measured after adenosine triphosphate (ATP) splitting by the contractile proteins was chemically inhibited. This tension-independent heat (TIH) (1 mJ/g wet weight) was used to calculate the total calcium (Ca) cycled in a muscle twitch by assuming that 87% of TIH was due to Ca2+ transport by the sarcoplasmic reticulum with a coupling ratio of 2 Ca2+/ATP split; the enthalpy of creatine phosphate hydrolysis buffering ATP was taken as -34 KJ/mol. The estimated Ca turnover per muscle twitch at 21 degrees C, 0.2 Hz pacing rate, and 2.5 mM Ca in the Krebs solution was approximately equal to 50 nmol/g wet weight. There was a tight positive correlation between TIH and mechanical activation during steady-state measurements but no correlation during the sharp increase in mechanical activation (treppe) when stimulation was resumed after a rest period. It is suggested that while total Ca cycling remains unchanged during the initial period of tension treppe, the free Ca2+ transient and mechanical activation increase sharply due to resaturation of high affinity Ca2+ buffers, other than troponin C, depleted of Ca2+ during the rest period.

Increase of intraerythrocytic fructose-1,6-diphosphate after incubation of whole human blood with fructose-1,6-diphosphate.

The incubation of whole blood with fructose-1,6-diphosphate (FDP) entails a statistically significant increase of intraerythrocytic FDP together with a decrease of blood glucose. The increase is not significant when equimolar amounts of fructose plus twice molar phosphate are used. The effect of FDP is decreased in the presence of an excess of oxygen. FDP added to the whole blood is removed from plasma by the activity of plasma enzymes and by the presence of blood cells as well. No specific interaction of FDP with plasma proteins seems to occur and the effects of FDP addition last longer than is compatible with the presence of FDP in the plasma.

Functional consequences of altered cardiac myosin isoenzymes.

This is a review of work dealing with the contribution of myocardial myosin isoenzymes to the performance of the heart muscle. Isoenzyme composition is altered in rabbit hearts by banding the pulmonary artery (increase in %V3) and thyroid hormone injection (increase in V1). Both treatments result in myocardial hypertrophy. The performance of the hearts rich in V3 or V1 myosin isoenzyme is assessed by simultaneous analysis of the myothermal and mechanical output of papillary muscles isolated from the right ventricle. Initial and tension dependent heart per tension time integral are decreased in the pressure overload and increased in the thyrotoxic hypertrophied hearts. Thus, the economy of isometric contraction is high in the hearts with the V3 myosin and low in the hearts with the V1 myosin. This change in performance is explained in terms of alterations in the cycling frequency and tension time integral of the myosin cross-bridge. In the pressure overload hearts the cross-bridge cycling frequency is decreased, while the tension time integral is increased. Conversely, in the thyrotoxic preparations the cycling frequency is increased and the tension time integral is decreased.

Macrophage lysozyme stimulation by fructose-1-6-diphosphate.

FDP produces an increase of serum lysozyme concentration which may be related to stimulation of the phagocytic activity. Mice macrophages in vitro produce extracellular and intracellular LSZ (lysozyme) and FDP (fructose-1-6-diphosphate) increases this production. Also in vivo FDP stimulates the macrophages intracellular lysozyme production. The toxic activity in vitro and the protection in vivo against Staphylococcus pyogenes after FDP administration can also be related to macrophage stimulation.

Human heart failure: determinants of ventricular dysfunction.

Thin muscle strips were obtained from non-failing (NF) and failing (dilated cardiomyopathy (DCM)) hearts, using a new harvesting and dissection technique. The strips were used to carry out a myothermal and mechanical analysis so that contractile and excitation coupling phenomena in the NF and failing (DCM-F) preparations can be compared. Peak isometric force and rate of relaxation in DCM-F were reduced 46% (p < 0.02) while time to peak tension was increased 14% (p < 0.03). Initial, tension dependent, tension independent and the rate of tension independent heat liberation were reduced 62-70% in DCM-F (p < 0.03). The crossbridge force-time integral (FTIXBr) was calculated from these measurements and was shown to increase 40% while the amount and rate of calcium cycled per beat was reduced 70%. As a result of these changes in the contractile and excitation-contraction coupling systems in DCM-F, the force-frequency relationship was significantly blunted while the power output was markedly reduced. These fundamental alterations account for the substantial ventricular dysfunction found in the dilated cardiomyopathic failing heart.

Work production and work absorption in muscle strips from vertebrate cardiac and insect flight muscle fibers.

Stretch activation, which underlies the ability of all striated muscles to do oscillatory work, is a prominent feature of both insect flight and vertebrate cardiac muscle. We have examined and compared work-producing and work-absorbing processes in skinned fibers of Drosophila flight muscle, mouse papillary muscle, and human ventricular strips. Using small amplitude sinusoidal length perturbation analysis, we distinguished viscoelastic properties attributable to crossbridge processes from those attributable to other structures of the sarcomere. Work-producing and work-absorbing processes were identified in Ca(2+)-activated fibers by deconvolving complex stiffness data. An 'active' work-producing process ("B"), attributed to crossbridge action, was identified, as were two work-absorbing processes, one attributable to crossbridge action ("C") and the other primarily to viscoelastic properties of parallel passive structures ("A"). At maximal Ca(2+)-activation (pCa 5, 27 degrees C), maximum net power output (processes A, B and C combined) occurs at a frequency of: 1.3 +/- 0.1 Hz for human, 10.9 +/- 2.2 Hz for mouse, and 226 +/- 9 Hz for fly, comparable to the resting heart rate of the human (1 Hz, 37 degrees C) and mouse (10 Hz, 37 degrees C) and to the wing beat frequency of the fruit fly (200 Hz, 22 degrees C). Process B maximal work production per myosin head is 7-11 x 10(-21) J per perturbation cycle, equivalent to approximately 2 kT of energy. Process C maximal work absorption is about the same magnitude. The equivalence suggests the possibility that a thermal ratchet type mechanism operates during small amplitude length perturbations. We speculate that there may be a survival advantage in having a mechanical energy dissipater (i.e., the C process) at work in muscles if they can be injuriously stretched by the system in which they operate.

The dependence of the latency relaxation on sarcomere length and other characteristics of isolated muscle fibres.

1. The latency relaxation has been examined in single fibres from frog striated muscle with particular attention given to its possible relation to Ca(2+) release during excitation-contraction coupling.2. Latency relaxations were recorded at 19-23 degrees C from massively stimulated (0.2 msec pulses) single fibres using two selected RCA 5734 transducer tubes in a bridge circuit.3. The depth of the latency relaxation has its full value when stimulus strength is between 40 and 400% above twitch threshold. Stronger stimuli reversibly diminish the latency relaxation.4. The variation in depth of latency relaxation with sarcomere length was found similar to that reported previously for multifibre preparations but in single fibres the peak of the curve consists of a plateau between sarcomere lengths of 2.8 mu and 3.2 mu.5. Sucrose hypertonicity increases the depth of the latency relaxation at sarcomere lengths below 2.8 mu but above this length it has either no effect or a depressant effect depending on the degree of hypertonicity.6. The maximal depth of the latency relaxation (measured at 3 mu) averaged 0.23% of the maximal tetanus tension (measured at 2.2 mu) and was strongly correlated (r = 0.87) with the latter in forty-five single fibres.7. The maximal depth of the latency relaxation is not correlated with the number of sarcomeres in series in a fibre.8. The results of this study are shown to fully support and extend Sandow's (1966) hypothesis that the latency relaxation is caused by release of activator Ca(2+) from the sarcoplasmic reticulum.

The response of the heart to stress: a biological view of myocardial adaptation and failure.

The response of the myocardium to persistent stress involves an increase in mass and a restructuring of the cellular and subcellular elements. The experiments described in this article are designed to test the hypothesis that the restructuring of the various systems (contractile, excitation-contraction coupling, recovery, etc.) that occurs in adaptive hypertrophy is a coordinated (matched) process. When the restructuring of the systems in response to stress occurs in an uncoordinated fashion, congestive heart failure results. In addition to controls, three heart models with normal pump performance are used (control, C; pressure overload, P; thyrotoxic, T; and pressure overload plus thyrotoxic, PT4) and one with inadequate pump performance (pressure overload plus thyrotoxic, PT2). In this analysis the contractile and excitation-contraction coupling systems are evaluated. The former is assessed by sensitive myothermal measurement of tension dependent heat (TDH) normalized for the isometric tension time integral (integral of Pdt). The latter is assessed from measurement of the time to peak isometric tension (TPT). The TDH/integral of Pdt (mu cal/g.cm.s) and TPT (ms) for the C, P, T, PT4, and PT2 hearts are 2.4, 1.8, 5.2, 5.1, and 0.1, mu cal/g.cm.s and 627, 816, 352, 484, and 465 ms, respectively. According to the coordination or matching hypothesis, if TDH/integral of Pdt is low, then TPT should be increased, or if TDH/integral of Pdt is high, then TPT should be decreased. Relative to control hearts, matched restructuring of the contractile and excitation-contraction coupling systems occurred for the P, T, and PT4 preparations. In these animals the hypertrophy has been adaptive and the pump performance is adequate.(ABSTRACT TRUNCATED AT 250 WORDS)

Determinants of energy utilization in the activated myocardium.

This is a review of work dealing with the effect of pressure overload and thryotoxic hypertrophy of rabbit hearts on the production of total activity related (TA) and initial (I) heats during isometric contraction. Pressure overload hypertrophy is produced by constricting the pulmonary artery with a spiral monel metal clip. Thyrotoxic hypertrophy is produced by 14 daily i.m. injections of 0.2 mg L-thyroxine per kilogram. Heat output is measured with Hill-type planar vacuum deposited bismuth and antimony thermopiles, and force is measured with a capacitance strain gauge. The pressure overload results in a depressed velocity of unloaded shortening, a depressed rate of isometric force development, and an increased time-to-peak tension. These changes are associated with a decreased myosin ATPase, a heart with no V1 myosin isoenzyme, and an increase in the economy of isometric force development (integral of Pdt/TA, integral of Pdt/I). The thyrotoxic hearts exhibit an increased velocity of shortening and rate of force development, and a decrease in time-to-peak tension. These changes are associated with an increase in myosin ATPase activity, a heart with increase in the V1 isoenzyme composition (88% V1), and a decrease in the economy of isometric force development (integral of Pdt/TA, integral of Pdt/I). The changes in the two types of hypertrophied hearts are interpreted in terms of altered cross-bridge cycling rates and changes in cross-bridge tension time integral as well as excitation contraction coupling phenomena. In the thyrotoxic hearts there is an increase in the economy of the recovery processes. Both types of hypertrophy are considered to be adaptive and involve the coordinated restructuring of the excitation-contraction, contractile, and recovery systems.

Heat, mechanics, and myosin ATPase in normal and hypertrophied heart muscle.

In this paper we review our previous work on the myothermic economy of isometric force production in compensated cardiac hypertrophy secondary to pulmonary artery constriction (pressure overload) and/or thyrotoxicosis (volume overload). Hypertrophy-induced changes in isotonic and isometric twitch mechanics are correlated with accompanying changes in actin-activated myosin ATPase and heat liberation. Heat measurements were made with rapid, high-sensitivity thermopiles on right ventricular papillary muscles from normal and hypertrophied rabbit hearts. Total activity-related heat was separated into initial and recovery heat. Initial heat was separated into a tension-dependent component (TDH) relating to cross-bridge activity, and a tension-independent component (TIH) relating to excitation-contraction coupling. There were oppositely directed changes in most parameters studied in pressure overload hypertrophy (P) as compared with thyrotoxic hypertrophy (T). Thus, in P there was depression (30-50% in the rate of isometric force production, mechanical Vmax, TDH and TDH rate, myosin ATPase, TIH, and prolongation in time-to-peak twitch tension, whereas in T all parameters were oppositely changed except for no change in TIH. Thyrotoxicosis following pressure overload reversed the P-induced changes in all parameters. There was a direct, linear relation between in vitro actin-activated myosin ATPase and in vivo TDH. However, TDH per unit twitch tension or tension-time integral varied inversely with ATPase, making force production more economical than normal in P muscles and less economical than normal in T muscles. These cellular changes beneficially equip P hearts for slow, high-pressure, economical pumping the T hearts for fast, high-volume, uneconomical pumping. The differences are similar to those between slow and fast skeletal muscle and between neonatal and adult skeletal muscle. The mechanism of these changes is discussed in terms of an enzyme kinetic scheme of chemomechanical coupling in actomyosin interaction.

Myocardial adaptation to stress from the viewpoint of evolution and development.

Myocardial hypertrophy, which results from the increase in demand placed on the heart, does not involve the simple enlargement of the myocardial cell. Instead, the cells are restructured in a manner that depends on the nature of the stress applied. The cellular reorganization involves the contractile, EC coupling, and recovery systems. In the remodeling of each of these systems, a repertoire of functional adaptation is used which bears a remarkable similarity to the changes seen in the differentiation of species and the development of muscle type. Thus, in pressure overload hypertrophy, where the maintenance of persistent high pressure in a necessity, the contractile system of the muscle is slower and more economical. This is similar to the specialization seen in the tortoise or in slow muscles like the rat soleus. In thyrotoxic hypertrophy, where the demand is to move the blood at high velocity, the muscle is fast and less economical. This is similar to the specialization seen in the frog sartorius or rat EDL. The restructuring involves specific molecular changes that uniquely adapt the muscles for the tasks at hand.

Activation heat and latency relaxation in relation to calcium movement in skeletal and cardiac muscle.

Measurements of activation heat, initial heat, twitch tension, and latency relaxation were made using thin-layer, vacuum-deposited thermopiles and isometric force transducers, respectively. Experiments were performed on frog skeletal muscle fiber bundles and on rabbit right ventricular papillary muscles at 0, 15, ans 21 degrees C in normal and 1.75X to 2.5X mannitol hyperosmotic bathing solutions. In skeletal muscle, activation heat, obtained by stretching to zero overlap, was only slightly affected by 1.75X hyperosmotic solution and consisted of a fast and a slow component. Both components have a refractory period and a relatively refractory period which can be demonstrated by double pulse stimulation. The twitch potentiators Zn2+ and caffeine increase the total activation heat and the magnitude and rate of the fast component. The temporal relation between the latency relaxation and activation heat is demonstrated. The latency relaxation is independent of the number of sarcomeres in series in a muscle. Activation heat and latency relaxation records from heart muscle are obtained in 2.5X hyperosmotic bathing solution. A model of excitation--contraction coupling is presented which indicates that (1) the downstroke of the latency relaxation monitors the functioning of the Ca2+-permeability or debinding mechanism in the terminal cisternae, (2) the fast component of activation heat monitors the amount of Ca2+ bound to troponin C, and (3) the total amplitude of activation heat is a measure of the total quantity of Ca2+ cycled in a twitch.

The partitioning of altered mechanics in hypertrophied heart muscle between the sarcoplasmic reticulum and the contractile apparatus by means of myothermal measurements.

Cardiac hypertrophy in the rabbit, secondary to pulmonary artery stenosis, results in a decrease in unloaded shortening velocity (Vmax) and maximum rate of isometric force development (dP/dtmax), while the peak isometric twitch tension is unchanged and time to peak tension (TPT) is increased. The principle hypothesis used to explain these results involve 1) slowing of myosin cross bridge movement as reflected in depressed myosin ATPase activity and 2) changes in excitation contraction coupling phenomena resulting in changes in intracellular Ca++ movement. Ca++ and actin activated myosin ATPase from the hypertrophied (H) muscles is depressed by 30%. Total initial heat, tension dependent heat and tension independent heat are depressed in H muscles by 57, 56, and 61% respectively. The rate of tension independent heat production in H preparations is depressed by 66%. From these data it is concluded that 61% of the depression in Vmax could be accounted for by the alteration in myosin with the reminder attributable to changes in EC coupling. Increased TPT can be accounted for by the change in rate of Ca++ flux as indicated by the alterated rate of tension independent heat evolution.

Tension-independent heat in rabbit papillary muscle.

1. Heat and force were measured from isometrically contracting (0.2 Hz) rabbit papillary muscles at 21 degrees C during a single contraction-relaxation cycle using antimony-bismuth thermopiles and a capacitance force transducer. 2. Tension-independent heat (TIH) associated with excitation-contraction coupling was isolated from the initial heat by eliminating tension and tension-dependent heat with a Krebs-Ringer solution containing 2,3-butanedione monoxime (BDM) and mannitol. 3. A strategy for testing the validity of this new method for measuring TIH in heart muscle is described and the test confirms that the BDM-hypertonic solution partitioning method properly estimates the magnitude of the TIH component of initial heat. 4. TIH at the time of complete mechanical relaxation is 1.00 +/- 0.17 mJ/g wet weight and the data suggest that calcium cycling is complete by this time. Conversion of TIH to calcium cycled, assuming that 87% of TIH is due to calcium pumping by the sarcoplasmic reticulum, indicates that approximately 52 nmol calcium/g wet weight are required to support a single cycle of mechanical activity (0.2 Hz, 21 degrees C). 5. The length and frequency dependence of excitation-contraction coupling were demonstrated. TIH is reduced by shortening muscle length and by increasing the interval between stimuli. These steady-state data suggest that only a portion (approximately 40%) of TIH is directly related to activation of the contractile apparatus. 6. TIH in the first twitch following a 45 min rest period is significantly reduced by approximately 30%. 7. With subsequent twitches in the positive treppe following the rest period, TIH does not increase as steeply as expected suggesting that tension rise in twitches 1-10 may be modulated by competitive binding of calcium rather than increased calcium delivery.

Heart muscle mechanics.

The goal implicit in the research reviewed above is to describe the contractile behavior of heart muscle in terms of crossbridge and filament behavior. It is necessary to elucidate these details in cardiac muscle because of the distinct biochemical differences between skeletal and cardiac myosin. As is evident in this review, significant advances have been made toward describing unique mechanical properties of cardiac muscle crossbridges. Several major problems now require attention: (a) Activation parameters are labile, making mechanical measurements sensitive to measurement perturbation; (b) significant structural inhomogeneities at the cellular and sarcomere level prevent precise assignment of externally measured force to internal structures (force generators, passive elements) within whole cardiac muscle and individual cells; (c) high resting stiffness and forces of poorly understood origin and properties confound attempts to interpret force measurements and dynamics. The differences between heart and skeletal muscle myosin may provide the means for identifying structural counterparts of the Huxley-Simmons model (33); they may also be useful in evaluating the electrostatic and quantum-mechanical models.