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Results of in vitro contracture tests for the diagnosis of malignant hyperthermia susceptibility in monozygote twins.

BACKGROUND: Malignant hyperthermia susceptibility is a pharmacogenetic disorder in which susceptible individuals may develop a potentially life-threatening hypermetabolism when exposed to certain anaesthetic agents. The most common diagnostic method is the in vitro contracture test (IVCT) of skeletal muscle biopsies. There is a wide variation in the size of contractures between susceptible individuals and the reproducibility of the test in humans has not been evaluated. METHODS: We have performed the IVCT in 4 monozygote pairs of twins, which gave us on opportunity to study the reproducibility. RESULTS: The clinical diagnoses were consistent in all twin pairs, although slight differences in contractures and thresholds were seen. CONCLUSION: In this material the reproducibility of the IVCT was found to be satisfactory.

Child↗

Bladder neck contracture following prostatectomy.

In an 11-year period, 49 patients developed bladder neck contracture after prostatectomy--an incidence of 0.86%. This complication was found to be more common after resection of small fibrous hyperplastic prostates. The best results for treatment of the contracture were obtained after bladder neck incision, which gave a 9% incidence of recurrence; after transurethral resection of the bladder neck contracture the recurrence rate was 46%, and after bladder neck dilatation it rose to 100%.

Aged↗

Two-stage correction of depressed glabella and nasal root scar contracture utilizing subcutaneous tissue advancement flaps and a layered soft tissue/procerus muscle transposition flap.

BACKGROUND: A postoperative nasal root and glabellar forehead deformity comprised both vertical and horizontal elements, with scar contracture and fixation across the left medial nose base, extending into the adjacent superior canthal region. METHODS: A two-stage reconstruction was performed, initially correcting the vertical centrally placed forehead depression by dual subcutaneous flaps advanced centrally and joined. The horizontal nasal root contracture was repaired at a second stage utilizing a procerus muscle transposition flap elevated on the ipsilateral deep contracture side, imbricated in serpentine fashion on itself and sutured into the depression over the scar base. RESULT: Significant improvement followed. CONCLUSION: The careful and precise placement of subcutaneous tissue and muscle can serve to assist in filling certain deep defects and in elevating scar lines.

Aged↗

Effect of external sodium substitution on potassium contractures of mammalian muscles: possible involvement of sarcolemma-bound calcium and Na+-Ca2+ exchange.

Isometric tension of K+ contractures, membrane potential, internal Na+ activity (alpha iNa) and intracellular pH (pHi) have been measured in vitro under conditions which modify sarcolemma-bound calcium and the activity of the Na+-Ca2+ exchange of normal mammalian soleus muscle. In the absence of external Na+ and for a given high external K+, the maximum amplitude of the contracture was increased (Na+ replaced by TEA+) or decreased (Na+ replaced by Li+ or Cs+) compared with that obtained in the presence of Na+. Replacement of external Na+ by another monovalent cation (TEA+) did not induce any change of pHi. However the concomitant decrease of alpha iNa was related to a Na+-Ca2+ exchange across the sarcolemmal membrane. The data suggest that in soleus muscle, a sarcolemmal calcium pool is involved during the development of K+ contractures.

Animals↗

The contracture produced by sodium removal in the non-pregnant rat myometrium.

1. Mechanical responses to Na removal were investigated in the circular and longitudinal muscles of the non-pregnant rat myometrium at 35 degrees C. In both muscles, reduction of the external Na concentration to less than 20 mM produced an initial acceleration of phasic contractions and a sustained tonic contracture. No difference was found with different Na substitutes (Tris-hydroxymethyl aminomethane, choline, dimethyl diethanol ammonium). However, when Mg was substituted for Na, only the tonic contracture was produced without the phasic contractions. 2. Readmission of 5-10 mM-Na, after exposure to Na-free solution, relaxed the contracture produced by Na removal. The degree of relaxation was dependent on the Na concentration readmitted and on the period of pre-treatment with Na-free solution, being stronger with longer pre-treatment. 3. In the presence of Na, excess Ca failed to increase the muscle tone. In the absence of Na, the tension development was closely related to the external Ca concentration up to 20 mM. In the absence of both Ca and Na, some tension remained. Even after pre-treatment with Ca-free solution containing 0.1-0.5 mM EGTA, removal of Na caused some mechanical response. A similar small tension development was observed when Na removal was repeated during prolonged absence of external Ca for more than 3 h. 4. Verapamil (2 X 10(-4) M) markedly suppressed the response to Na removal, but it did not block it, either in the presence or in the absence of Ca. Ouabain (10(-3) M) in the presence of verapamil potentiated the early phasic component of the response to Na removal, but the tonic component was little affected or even slightly reduced. 5. The results indicate that there are three components in the mechanical response to Na removal: the phasic and tonic components, which are highly Ca-dependent, and the third small tonic component, which is independent of external Ca. Most of the phasic and tonic responses seem to be due to an increase in Ca permeability, but this may be secondary to membrane depolarization. A Na-Ca exchange mechanism is also considered to contribute to the transient phase of the response to Na removal and to Na readmission.

Animals↗

Contracture and catecholamines in mammalian myocardium.

Contractures induced by KCl are produced in cat ventricular muscle after depletion of catecholamine stores by previous treatment with reserpine or by selective blocking of adrenergic receptor sites with propranalol. Contractures are reproducible for 2 to 3 hours and are markedly depressed by the addition of epinephrine. The relaxation of contracture induced by epinephrine parallels the eftect of this compound of shortening the duration of the normal twitch. The failure of the normal mammalian myocardium to maintain tension in solutions of high KCI concentration appears to be related to the endogenous amounts of catecholamines in the tissue.

Animals↗

Ba2+ ions block K+-induced contractures by antagonizing K+-induced membrane depolarization in frog skeletal muscle fibres.

The effects of Ba2+ ions on twitches, K+-induced contractures, and on intracellularly recorded membrane potentials (Em) and depolarizations of frog skeletal muscle fibres were investigated. Exposure of toe muscles to choline--Ringer's solution with 10(-3) M Ba2+ with Ca2+ (1.08 mM) eliminated or very greatly reduced contractures produced by 60 mM K+. In contrast, not only did the same concentration of Ba2+ ions fail to depress the twitch tension of isolated semitendinosus fibres when added to Ringer's with Ca2+, but it even restored twitches that had been eliminated in a zero Ca2+ Ringer's solution. The resting Em of sartorius muscle fibres in choline--Ringer's solution was reduced about 20 mV by 10(-3) M Ba2+. This Ba2+ ion concentration also antagonized the K+-induced depolarization. Thus in the presence of 1 mM Ba2+, 20 mM K+ hyperpolarized rather than depolarized the fibres and 60 or 123 mM K+ produced only very slowly developing, small depolarizations. These results suggest that the loss of the K+-induced contracture in choline-Ringer's caused by Ba2+ ions is due to an inhibition of the K+-induced depolarization. The latter result is consistent with previous findings of other workers that Ba2+ ions block membrane K+ channels.

Action Potentials↗

External calcium and contractile activation during potassium contractures in twitch muscle fibres of the frog.

Effects of external Ca2+ concentration reduction on the amplitude and time course of K+ contractures were studied in single muscle fibres. The resting potential, effective resistance, threshold for the Na current, action potential and K+-induced depolarizations did not change when 1.8 mM Ca2+ was replaced by 3 mM Mg2+ (3--6 microM Ca2+). Identical results were obtained after the addition of 5 mM EGTA (less than or equal to 10(-9) M Ca2+; Ca-free saline). The rate of tension development during the initial phase of K+ contractures was independent of external Ca2+ while the amplitude, the duration, and the time constant of spontaneous relaxation decreased progressively as Ca2+ concentration was diminished. The activation curve shifted by 3--5 mV towards more positive potentials while the inactivation curve shifted by 16--18 mV in the opposite direction and both curves became steeper in Ca-free saline. External Ca2+ may play a role in excitation--contraction coupling during K contractures either via the inward Ca current or via specific interactions between external Ca2+ ions and the coupling mechanism or both.

Action Potentials↗

Nitrendipine blocks high potassium contractures but not twitches in rat skeletal muscle.

The effects of the organic calcium channel blocker nitrendipine was tested on electrically evoked twitches and on potassium depolarization-induced contractures of rat lumbricalis muscles. Nitrendipine (10(-7) to 5 X 10(-5) M) blocked only the potassium contractures. It was concluded that blocking calcium uptake through the slow voltage-sensitive calcium channels during potassium depolarization blocks the mechanical response of the muscle. Thus extracellular calcium ions are required for the excitation-contraction (E-C) coupling during depolarization contractures. On the other hand, electrically evoked twitches were not affected by nitrendipine; therefore, extracellular calcium ions entering via the slow voltage-sensitive channels are not required for E-C coupling during the twitch.

Animals↗

Submaximal sodium-lack contractures in rapidly perfused frog ventricular strips.

Rapid perfusion of strips of frog ventricular muscle (width less than or equal to 8 mm) by sodium-free Ringer yielded contractures with very fast rates of rise (half time approximately equal to 2 s) and flat plateaus. Tension in lithium Ringer was 0.64 (0.38-0.77) of the maximal tension the muscle could produce [i.e., for an extracellular potassium concentration ([K]o) of 160 mM, and an [Na]o of 0 mM at less than 15 degrees C], showing that intracellular calcium concentration ([Ca]i) in zero [Na]o and normal [K]o was insufficient to saturate the myofilaments. Tension was related to [Ca]o in an S-shaped manner (apparent Km = 1.3 microM), was rapidly increased by electrical depolarization, high potassium, or low temperature, but was unaffected by up to 10(-5) M dihydroouabain. From these data was derived the working hypothesis that the value of [Ca]i during the contracture plateau is a steady-state value due to influx through a sodium-dependent mechanism and calcium uptake or efflux via a sodium-independent mechanism. Also, 1 microM dihydroouabain increased peak twitch tension by 60% but did not potentiate tension during the plateaus of contractures obtained by partial replacement of Nao. This result supports the working hypothesis and suggests that the potentiating effect of raising Nai may depend on the membrane potential (D. A. Eisner, W. J. Ledener, and R. D. Vaughan-Jones. J. Physiol. Lond. 335: 723-743, 1983).

Animals↗

Iodoacetate-induced contracture in rat skeletal muscle: possible role of ADP.

The effects of iodoacetic acid (IAA) and ischemic contraction were studied in rat extensor digitorum longus muscles. Ischemic contraction of IAA-treated muscles produced contracture. The onset of contracture was not associated with a change in sarcolemmal electrical properties or reduction in intracellular [ATP]; however, [creatine phosphate] was reduced by 75% and free [ADP] was increased by 665%. Continued stimulation of IAA-treated fibers resulted in depolarization, loss of membrane excitability, further depletion of creatine phosphate, and reduction in [ATP]. The effects seen in IAA-treated muscle did not appear to result from a direct action of IAA on the surface membrane, contractile proteins, or excitation-contraction coupling. The contractures in IAA-treated muscle may have resulted from increased Ca sensitivity of the contractile proteins, increased myoplasmic [Ca], or both. Both effects may have resulted from increased [ADP]. In addition, the reduced acidification during ischemic contraction of IAA-treated fibers compared with control fibers may have further increased the Ca sensitivity of IAA-treated fibers compared with controls.

Adenosine Diphosphate↗

Contracture of isolated rat heart cells on anaerobic to aerobic transition.

Adult rat heart myocytes prepared by collagenase perfusion show a progressive loss of adenylate energy charge and total adenine nucleotide as a function of time of anaerobic incubation in the absence of glucose. Re-aeration of the rod-shaped anaerobic cells produces a population of viable rounded cells in hypercontracture. The round cells show extensive morphological dislocations but remain metabolically competent in that they 1) restore adenosine 5'-triphosphate levels to the extent permitted by the depleted adenine nucleotide pool: 2) reestablish a low Na+-K+ ratio; and 3) restore creatine phosphate to 73% of control. The hypercontracture on re-aeration of anaerobic myocytes closely resembles an analogous contracture of heart cells in situ produced when hypoxic perfused hearts are reoxygenated, the so-called "oxygen paradox." Both processes are eliminated by inclusion of glucose during the anaerobic phase and by inhibitors of respiration and uncouplers of oxidative phosphorylation added before reoxygenation. Mitochondria in the hypercontracted myocytes retain high acceptor control ratios. Contracture on re-aeration occurs to nearly the same extent in the presence of either mM Ca2+ or 0.1 mM EGTA. Contracture appears related to dislocations in intracellular Ca metabolism that result from the declining energy charge and depleted nucleotide pool produced during anoxic incubation.

Adenine Nucleotides↗

Isotonic length transient of cat heart muscle in Ba2+-induced contracture.

To characterize mechanical properties of activated heart muscle, the length response to step (4 ms) decrease in tension in Ba2+-induced contracture in kitten papillary muscle was analyzed. The amplitudes of tension steps were varied at the maximum contracture level (Tc), different initial muscle lengths [0.90 Lmax to Lmax], and different temperatures (20-35 degrees C). When the tension decrease was less than 0.7 Tc, the length response comprised four different phases. The amplitude of shortening in the second phase, after the initial rapid shortening in the first phase, increased up to approximately 3.5% of the initial muscle length quite linearly with increasing amplitude of tension reduction. The amount of lengthening in the third phase, however, increased to a maximum (up to approximately 1.4% initial length) at the tension reduction of approximately 0.4 Tc. The third phase of lengthening was larger at shorter initial muscle length. Increasing temperature markedly decreased the amplitude and shortened the duration of the length response in the second and third phase independently of initial muscle length. Unlike that of the muscle in contracture, the length response of the resting and of the rigor muscle changed in a nearly stepwise fashion. One explanation of these phenomena is that the transient length responses following a rapid tension reduction are mostly determined by the kinetics of the attachment and detachment of cross bridges between actin and myosin filaments.

Animals↗

Mechanism of ischemic contracture in ferret hearts: relative roles of [Ca2+]i elevation and ATP depletion.

When coronary perfusion is interrupted, the diastolic force generated by the myocardium first falls but eventually increases. The delayed rise in force, ischemic contracture, has been attributed either to ATP depletion or to elevation of the intracellular free calcium concentration ([Ca2+]i). To distinguish between these possibilities, we measured [Ca2+]i and ATP concentration [( ATP]) in ferret hearts using nuclear magnetic resonance (NMR) spectroscopy. Mean time-average [Ca2+]i and [ATP] equaled 0.25 microM and 2.7 mumol/g wet wt, respectively, under control perfusion conditions. [Ca2+]i increased and [ATP] fell during total global ischemia. Although [Ca2+]i exceeded the usual systolic levels of 1.7 microM within 20-25 min of ischemia and reached a steady level between 2 and 3 microM by 30-35 min, force only began to rise after 40 min. In contrast, the time required for [ATP] to fall to less than 10% of control levels coincided closely with the onset of contracture. Ischemia in the presence of iodoacetate, an inhibitor of glycolysis, led to a precipitous fall in [ATP] and a concomitant rise in force, both of which preceded any elevation of [Ca2+]i. Thus changes in [Ca2+]i are neither sufficient nor necessary for the initiation of ischemic contracture. We conclude that ATP depletion is primary and that the rise in resting force reflects the formation of rigor cross bridges.

Adenosine Triphosphate↗

Effect of lidocaine on contracture, intracellular sodium, and pH in ischemic rat hearts.

The relationships among intracellular Na concentration ([Na+]i), intracellular pH, [ATP], and contracture during global ischemia were studied in isolated, perfused rat hearts. Intracellular Na was monitored by 23Na nuclear magnetic resonance (NMR) spectroscopy using thulium 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra(methylene phosphonate) (TmDOTP5-) as the paramagnetic shift reagent. High-energy phosphates and pH were monitored under the same conditions using 31P-NMR spectroscopy. Lidocaine (130 microM), a class IB fast Na channel blocker known to protect ischemic myocardium, prolonged the time to contracture in both unpaced and paced hearts (240 beats/min). After 10 min of global ischemia in paced hearts, [Na+]i was lower in the lidocaine-treated group compared with untreated hearts. The addition of lidocaine also significantly attenuated the depletion of ATP as well as development of intracellular acidosis. At the time of contracture, however, there was no difference in [Na+]i or pH between the two groups. Interestingly, the effect of lidocaine on Na+i accumulation during ischemia was manifested during the first 5-10 min of ischemia, while its effect on pH occurred after 9 min. This finding suggests that a mechanism other than the Na-H exchanger may play a role in the accumulation of Na+i early in the course of ischemia.

Animals↗

Rapid cooling contractures in rat skinned myocardium: effects of isoflurane and halothane.

Rapid cooling contractures (RCCs) have been elicited in rat ventricular cardiac muscle skinned by saponin (50 micrograms/ml). The size and shape of the RCC in skinned cardiac muscle were similar to those observed in intact cardiac muscle. The ATP-dependent Ca2-uptake pump in the sarcoplasmic reticulum (SR) was active at low temperature and could efficiently load the SR with Ca2+ even at 3 degrees C. Halothane reduced both RCC and caffeine contracture in a dose-dependent way when halothane treatment was applied before either RCC or caffeine. This action is consistent with halothane-induced depletion of the SR Ca2+. Under similar conditions, isoflurane inhibited RCC but had little effect on the caffeine contracture. This may suggest that rapid cooling and caffeine have different modes of action on the SR Ca(2+)-release channel. Our results provide further strong supporting evidence for differential inhibitory actions on a key intracellular organelle in cardiac muscle by two vapor anesthetic agents.

Anesthetics, Inhalation↗

Energy expenditure by Ba(2+) contracture in rat ventricular slices derives from cross-bridge cycling.

To clarify the energy-expenditure mechanism during Ba(2+) contracture of mechanically unloaded rat left ventricular (LV) slices, we measured myocardial O(2) consumption (VO(2)) of quiescent slices in Ca(2+)-free Tyrode solution and VO(2) during Ba(2+) contracture by substituting Ca(2+) with Ba(2+). We then investigated the effects of cyclopiazonic acid (CPA) and 2,3-butanedione monoxime (BDM) on the Ba(2+) contracture VO(2). The Ca(2+)-free VO(2) corresponds to that of basal metabolism (2.32 +/- 0.53 ml O(2). min(-1). 100 g LV(-1)). Ba(2+) increased the VO(2) in a dose-dependent manner (from 0.3 to 3.0 mmol/l) from 110 to 150% of basal metabolic VO(2). Blockade of the sarcoplasmic reticulum (SR) Ca(2+) pump by CPA (10 micromol/l) did not at all decrease the Ba(2+)-activated VO(2). BDM (5 mmol/l), which specifically inhibits cross-bridge cycling, reduced the Ba(2+)activated VO(2) almost to basal metabolic VO(2). These energetic results revealed that the Ba(2+)-activated VO(2) was used for the cross-bridge cycling but not for the Ca(2+) handling by the SR Ca(2+) pump.

Animals↗

Increased myometrial contracture frequency at 96-140 days accelerates fetal cardiovascular maturation.

Fetal cardiovascular responses to an altered intrauterine environment of increased myometrial contractures induced by oxytocin (OT) pulses to the ewe over the final 50 days of gestation were studied in chronically instrumented sheep. Ewes received saline (Cntl) or long-term OT treatment (LTOT, 600 microU x kg(-1) x min(-1) in 5-min pulses every 20 min) from 96 days gestational age. Fetal baroreflex responses to sodium nitroprusside (SNP) and phenylephrine (PE) were studied at 133 days gestation. OT increased contractures in LTOT ewes. Fetal blood pressure (FBP) was higher, and fetal heart rate (FHR) and slope of daily change in FBP and FHR were lower in LTOT fetuses. Fetal SNP-induced hypotension resulted in a narrow R-R interval variation range in LTOT fetuses; Cntl fetuses showed early breakdown in compensation. Baroreflex response slope during PE-induced fetal hypertension was lower in LTOT than in Cntl fetuses. Although the cortisol-to-ACTH ratio was lower in LTOT fetuses, fetal plasma ACTH and cortisol changes were similar in control and LTOT fetuses. We hypothesize that contracture-induced alterations in the intrauterine environment accelerate fetal cardiovascular development through mild hypoxemia, repetitive fetal pituitary-adrenal stimulation, and/or physical stimulation.

Animals↗