Caffeine contractures, twitch characteristics and the threshold for Ca(2+)-induced Ca2+ release in skeletal muscle from horses with chronic intermittent rhabdomyolysis.

Muscle from horses with intermittent exercise associated rhabdomyolysis was examined to determine if calcium regulation was abnormal. In vitro studies on semimembranosus muscle fibre bundles showed the time to 50 per cent relaxation of caffeine-induced contractures was shorter and the electrically elicited twitch longer in horses with exercise associated rhabdomyolysis. Substitution of strontium for calcium eliminated the difference in caffeine contracture between the normal and rhabdomyolysis horses. The threshold of calcium-induced calcium release was lower than normal in terminal cisternae-containing fractions of muscle from horses with rhabdomyolysis. Thoroughbreds with rhabdomyolysis had a shorter time to peak twitch tension than standardbreds, and normal thoroughbreds had a shorter caffeine contracture than normal standardbreds. There was no difference in fibre typing between breeds or groups. Either no histological changes or low grade to moderate degenerative myopathy was seen in muscle from horses with rhabdomyolysis. These results suggest horses with intermittent exercise associated rhabdomyolysis have abnormal calcium regulation.

Triglycerides, not phospholipids, are the source of elevated free fatty acids in muscle from patients susceptible to malignant hyperthermia.

In the absence of halothane challenge, incubates of skeletal whole-muscle homogenates from malignant-hyperthermia-susceptible humans and pigs exhibit greater free fatty acid production than controls, which has been attributed to elevated phospholipase A2 activity. The present study examines lipid profiles of human muscle (vastus lateralis) prior to halothane challenge, relating the lipid profiles to the magnitude of halothane contracture response in muscle from the same biopsy. No differences in cholesterol, phospholipids or free fatty acids were observed among preparations exhibiting low, moderate or strong responses to halothane. Two fatty acids associated with triglycerides (palmitoleic and oleic) were significantly (P less than 0.05) lower in muscle demonstrating a strong response to halothane. These results do not support altered phospholipase A2 activity as a defect in malignant hyperthermia, but rather support an enhanced turnover of triglycerides in biopsied skeletal muscle from MH-susceptible humans.

Effects of droperidol, haloperidol and ketamine on halothane, succinylcholine and caffeine contractures: implications for malignant hyperthermia.

The effects of two structurally similar butyrophenones (droperidol and haloperidol) and ketamine were evaluated in an in vitro system to determine their potential for eliciting or exacerbating an episode of malignant hyperthermia. Muscle strips from patients referred for diagnostic testing for malignant hyperthermia and muscle strips from the rat diaphragm were exposed to droperidol, haloperidol, or ketamine prior to challenge with halothane, succinylcholine or caffeine. If any agent augmented the contracture response to the malignant hyperthermia triggering or diagnostic agents, then the agent was considered unsafe for use in malignant hyperthermia susceptible patients. Droperidol 10 mumol/l and ketamine 100 mumol/l did not induce contractures in human or rat skeletal muscle when added alone, nor did they augment halothane, succinylcholine or caffeine contractures. These agents appear to be safe for use in patients susceptible to malignant hyperthermia. In contrast, haloperidol 10 mumol/l augmented the response to succinylcholine about 1.5-fold and may be contraindicated in MH susceptibles.

Effect of phenytoin on the clinical signs and in vitro muscle twitch characteristics in horses with chronic intermittent rhabdomyolysis and myotonia.

In vitro twitch characteristics of the semimembranosus muscle were evaluated in 9 clinically normal horses, in 15 horses with chronic intermittent rhabdomyolysis (CIR) and in 2 horses with myotonia. Effects of phenytoin on in vitro muscle twitch and clinical signs of CIR and myotonia were evaluated in these same horses. Times to 90% relaxation were prolonged in the horses with CIR (mean +/- SEM, 186 +/- 5.9 ms) and in 2 horses with myotonia (197 and 177 ms) compared with those in clinically normal horses (mean +/- SEM, 146 +/- 2.1 ms). Horses with CIR also had significantly (P less than 0.05) longer times to 50% relaxation, compared with clinically normal horses. In the group of horses with CIR, Standardbreds had significantly (P less than 0.05) longer times to 90% and 50% relaxation, compared with Thoroughbreds. Times to 100% peak tension did not differ among the groups. Administration of phenytoin directly into a muscle preparation bath solution had no effect on muscle twitch properties. After the initial muscle biopsy, phenytoin was administered orally for 7 to 10 days to 4 horses with CIR, 2 myotonic horses, and 2 clinically normal horses before repeat biopsy from the same site in the contralateral semimembranosus muscle. Times to 90% relaxation decreased from 197 and 177 ms to 144 and 126 ms, respectively, in the 2 myotonic horses, from a mean of 192 (+/- 9) ms to 170 (+/- 9) ms in the 4 horses with CIR and remained unchanged (154 and 140 ms before vs 155 and 139 ms after treatment) in the 2 clinically normal horses.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of varying concentrations of linoleic acid on alpha-adrenoceptor responses in spontaneously hypertensive rats.

The effect of increased intake of linoleic acid on the alpha-adrenergic system was assessed by safflower oil supplementation to spontaneously hypertensive rats. Linoleic acid-enriched intake at 5%, 15% and 30% by weight of total food intake for 12 wk was associated with a reduction in resting arterial blood pressure, while heart rate and heart to body weight ratios were similar to control group values. A dose-response analysis to norepinephrine bitartrate administered intravenously indicated a significant reduction in the vascular reactivity to this alpha-adrenergic agonist in all groups given linoleic acid. Direct assessment of alpha-adrenoceptor number (Bmax) and affinity (KD) in cardiac sarcolemma with [3H]-prazosin indicated that receptor binding properties were not affected by linoleic acid intake. Our results suggest that short-term linoleic acid supplementation in the established hypertensive state may lower blood pressure through effects upon alpha-adrenergic reactivity in vascular tissue, without associated effects in cardiac tissue.

Development of an animal model for investigating disparate myocardial effects of obesity and hypertension.

An experimental model for investigating the disparate effects of obesity and hypertension on the heart was developed by ligation of the aorta of male Sprague-Dawley rats made obese through ad libitum feeding. Experimental obesity was associated with an increased body fat and cardiac muscle mass, yet a normotensive systemic arterial pressure. Aortic ligation produced an elevated mean arterial pressure and resting heart rate, whereas body weight was similar to that of normotensive lean control rats. Obesity and hypertension together were associated with a significantly increased percent body fat, mean arterial pressure, and left ventricular mass compared with lean controls, whereas pressure and left ventricular weight were greater than those observed in rats with only obesity or hypertension. Cardiac adaptations corrected for body weight indicated that left ventricular weight increased as a function of body weight and body fat, but hypertension produced left ventricular adaptations independent of these variables. These initial studies indicate an additional contribution of hypertension to the left ventricular adaptations of obesity, and this model could therefore be used in future investigations concerning the cardiovascular effects of the simultaneous occurrence of these separate diseases.

Antilipolytic activity of viprostol, a transdermally active antihypertensive PGE2 analog.

Viprostol, a novel prostaglandin E2 congener, was assessed for in vitro antilipolytic activity in the spontaneously obese rat. In isolated epididymal adipocytes, viprostol exhibited a dose-dependent inhibition of catecholamine-stimulated lipolysis at concentrations ranging from 10 microM to 1 mM, but was ineffective at lower concentrations. Additionally, viprostol exhibited approximately 50% of the antilipolytic activity of naturally-occurring PGE1 and PGE2 at similar concentrations, but was as potent as PGF2 alpha. At 10 microM, viprostol inhibited maximum catecholamine-stimulated lipolysis by approximately 35% of the total, hormone-stimulated glycerol release. The results of these experiments indicate that viprostol exhibits antilipolytic activity in vitro, but is less potent than the naturally-occurring PGE's to which it is most closely related structurally.

Contracture induction by snake venom cardiotoxin in skeletal muscle from humans and rats.

Contracture responses to cardiotoxin (CTX) from Naja naja kaouthia venom were investigated in rat and human skeletal muscle of similar fiber type distribution to determine species differences in mechanism of action. Rat diaphragm strips and human vastus lateralis preparations were directly stimulated in a tissue bath. The calcium dependence of toxin action, synergism between CTX and phospholipase A2 (PLA2) activity and roles of Na+ + K+-ATPase activity and the sarcoplasmic reticulum Ca2+ stores in contracture induction were examined. The threshold of contracture to CTX was decreased in human and rat muscle when Sr2+ was substituted for Ca2+ in the bathing medium. In rat, but not in human muscle the threshold of contracture to CTX was decreased in a medium in which Ca2+ had been omitted. The decrease in contracture threshold may relate to toxin binding. The maximum height of contracture for preparations from humans, but not for those from rats was considerably depressed in a medium in which Ca2+ had been omitted. Exogenously added bee venom PLA2 acts synergistically with CTX in skeletal muscle in a manner similar to that in erythrocytes. Ouabain (100 microM) did not elicit contractures in any of the media tested nor affect CTX-induced contractures in Sr2+-containing medium. Dantrolene antagonized CTX-induced contractures, suggesting a role for Ca2+ derived from the sarcoplasmic reticulum in CTX action. The species difference in CTX action may reflect differences in the relative contribution of Ca2+ from the sarcolemma and sarcoplasmic reticulum to the contracture.

Hemodynamics of obesity: influence of pattern of adipose tissue cellularity.

Direct quantitation of blood flow with radioactive microspheres in conscious spontaneously obese rats indicated that the development of obesity was associated with an elevated cardiac output and stroke volume, a normotensive blood pressure, and a reduced total peripheral resistance when directly comparing obese rats with their lean counterparts. Obesity was also associated with increased blood flow and decreased regional vascular resistance in a variety of vascular beds, whereas cardiac index and total peripheral resistance per unit of body weight were similar between groups. When corrected for tissue weight, unique alterations in flow and resistance were observed in the adipose tissue. When expressed as resistance per organ, the greatest relative alterations in vascular resistance with the development of obesity also occurred in the adipose tissue. Furthermore, localized adipose tissue expansion through cellular hypertrophy was consistently associated with a different pattern of blood flow and vascular resistance than adipose tissue that expanded through both cellular hypertrophy and hyperplasia, implying an association between depot cellularity and its hemodynamic profile.

Myocardial adaptations to dietary sucrose in spontaneously hypertensive rats.

The effects of dietary sucrose upon specific myocardial and hemodynamic parameters were examined in spontaneously hypertensive rats (SHR). Following a 6 wk program, SHR consuming a supplement of 10% sucrose in the drinking water exhibited increased heart weight, heart mass, left ventricular free wall thickness and increased resting heart rate when compared to the hypertensive control group. Total caloric intake was similar between groups. Cardiac alpha 1-adrenoceptor density and affinity were also altered following sucrose feeding. These data suggest that modest intake of dietary sucrose is associated with cardiovascular adaptations that may further burden a heart already compromised by the presence of systemic hypertension.

Effect of contractile activity on rat skeletal muscle beta-adrenoceptor properties.

The effect of fiber type and endurance exercise training on skeletal muscle beta-adrenoceptor properties were assessed using a direct radioligand binding technique. Six separate muscles, composed of a variety of different fiber types, were examined in treadmill trained and sedentary rats. In trained animals, sarcolemmal preparations from heart and slow twitch soleus muscle exhibited a significantly greater receptor concentration than membranes from white fast twitch glycolytic fibers of the vastus lateralis. No significant changes were observed between trained and sedentary rat muscle beta-adrenoceptor density (beta max, fmole/mg protein) or affinity (Kd, nM) within each muscle type, despite significantly increased myocardial/body weight ratios and skeletal muscle enzyme adaptations associated with the exercise program. These results suggest that muscle beta-adrenoceptor properties may be influenced in part by the motor nerve innervation to that muscle, and are further discussed with respect to a possible relationship between exercise intensity and receptor regulation.

Effects of the antihypertensive prostaglandin analog CL 115,347 on cardiac output distribution in the spontaneously hypertensive rat.

The effects of CL 115,347, a novel prostaglandin E2 (PGE2) congener, on a variety of hemodynamic variables were examined in conscious spontaneously hypertensive rats. Within minutes of topical administration (0.03, 0.3, or 3.0 mg/kg), a dose-dependent decrease in mean arterial blood pressure (MABP) was observed. One hour following drug application, a stable, dose-related hypotensive effect was still apparent, with MABP from 18 to 33 mm Hg lower than control, predose values. Quantitation of blood flow using radioactive microspheres indicated that a significant increase in stroke volume and cardiac output (3.0 mg/kg only), as well as a reduction in total peripheral resistance, was associated with the administration of CL 115,347. Further analysis of regional blood flow implied that vasodilation in specific vascular beds may contribute importantly to the antihypertensive activity of this compound.

Alterations in cardiac composition with weight reduction in the obese rat.

Alterations in myocardial composition associated with obesity and weight reduction have been examined in the spontaneously obese rat. When compared to values obtained from obese animals, body weight reduction was associated with a significant decrease in body fat, heart weight, and absolute left ventricular mass. Compositional analysis indicated that protein, lipid, and water together accounted for approximately 98% of total heart weight, and that while each component decreased with weight reduction, decreased myocardial water content represented the largest contribution to the reduced total heart weight. These data therefore suggest that the cardiomegaly of obesity is contributed to by specific changes in myocardial composition which can be altered through body weight reduction.