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Experimental study of lymphatic contractility and its clinical importance.

Lymphatic contractility in rats was studied, and changes in lymphatic contractility were recorded under the influence of various factors. We suggest that this contraction is not only the essential driving force in lymph drainage, but also that it plays an important role in the effect resulting from external forces. It was proved that there are alpha- and beta-receptors on the smooth muscle of the lymphatics. Under the control of neurohumoral factors, alpha and beta receptors regulate the lymphatic motion coordinately. We recommend that the lymphatics that still have contractile function be selected for lymphaticovenous anastomoses or other microlymphatic operations for treating lymphedema. Diluted epinephrine is dropped onto the local lymphatic to identify its contractile function.

Animals↗

[Gastric emptying of fluids and gallbladder contraction in non-ulcer dyspepsia].

Non-ulcer dyspepsia is a frequent clinical entity characterized by chronic upper gastrointestinal symptoms without organic lesions by radiology, sonography or endoscopy. We studied simultaneously the rate of gastric emptying and gallbladder contraction in 10 patients with non-ulcer dyspepsia and in 10 healthy controls after ingestion of 500 cc of isotonic saline, and 500 cc of a liquid mixed meal, hypercaloric and hyperosmotic. The measurements were done by direct real-time ultrasonic imaging in fasting and were repeated at regular intervals (15-30 minutes) after the liquid meal, until complete gastric emptying was established. We found with the liquid mixed meal, the patients with non-ulcer dyspepsia had a significant delay in gastric emptying and gallbladder hypokinesis with faulty contraction and slow refilling. Dynamic ultrasonographic studies are useful in patients with non-ulcer dyspepsia.

Adult↗

Inhibitory actions of laxatives on motility and water and electrolyte transport in the gastrointestinal tract.

Castor oil and magnesium sulfate were studied for their effects on gastrointestinal contractile activity in vivo. Ricinoleic acid, the active ingredient in castor oil, magnesium sulfate and mannitol were studied and compared for their effects on net water and electrolyte absorption in vitro. Extraluminal strain gauge transducers were implanted in dogs and used to monitor the circular smooth muscle activity of the antrum, duodenum, ileum and colon after water, castor oil or a 30 percent solution of magnesium sulfate. Substances were tested during the interdigestive (fasted) and digestive (fed) states. Decreases in total activity were found for both cathartics in the antrum and ileum. Further analysis revealed that decreases in the ileum occurred primarily by a decrease in contractile rate, whereas antral decreases could be attributed primarily to a decrease in force per contraction. Proximal colonic activity tended to decrease after laxatives and feeding. The effects of ricinoleic acid, isotonic and hypertonic solutions of magnesium sulfate and mannitol on net water and electrolyte absorption were tested on everted segments of hamster jejunum. Sodium ricinoleate (2.0 mM) reduced net water transport by 48 percent (P smaller than .01). Magnesium sulfate, like mannitol, only reduced net water absorption when present as a component of a hypertonic mucosal solution. The results suggest that both inhibition of water absorption and reduced circular smooth muscle activity may be important factors in castor oil- and magnesium sulfate-induced catharsis.

Animals↗

Passive sensitization of human bronchi augments smooth muscle shortening velocity and capacity.

We assessed whether incubation with human serum from atopic individuals containing high concentrations of immunoglobulin E (IgE) causes augmentation of maximal contraction of human bronchial smooth muscle from non-atopic subjects in vitro. Bronchi were obtained from eight patients undergoing lung resection, and force-velocity relationships were determined for eight pairs of epithelium-intact bronchial rings of generations 6-7 using an electromagnetic lever system, which allowed isotonic shortening when load-clamps [from 0 to maximal isometric force (P0)] were applied at specific times after onset of contraction. Contractions were elicited by supramaximal electrical field stimulation (50 Hz, 10 s train duration, 25 V). Optimal length (Lo) for each tissue was determined during equilibration. After resection, tissues were sensitized passively with human sera containing high titers (> 1,000 U/ml) of IgE by incubation for 16 h at 20 degrees C. Maximal shortening velocity (Vmax) was increased for passively sensitized bronchi [0.1150 +/- 0.0240 1/2 circumferences/s (1/2Cir/s)] compared with sham-sensitized bronchi [0.0731 +/- 0.0152 1/2Cir/s, P = 0.038]. Similarly, maximal shortening (delta Lmax) was augmented in sensitized bronchial rings (11.27 +/- 1.80 %Lo) compared with sham-sensitized tissues (8.19 +/- 1.39 %Lo, P = 0.012). However, P0 did not differ between sensitized (122.5 +/- 24.4 mN/cm2) compared with sham-sensitized tissues (138.4 +/- 32.1 mN/cm2, P = 0.642). Our data are the first demonstration that Vmax and delta Lmax are augmented in sensitized but not challenged human bronchial rings after passive sensitization using human serum containing high concentrations of IgE.

Bronchi↗

Muscle training by means of combined electrical stimulation and volitional contraction.

INTRODUCTION: A variety of techniques have been used with variable success in an attempt to reverse the loss of muscle strength, bulk, and endurance that occurs during spaceflight. This study was designed to evaluate a new "hybrid" approach that uses the resistance provided by an electrically stimulated antagonist muscle to resist the volitional contraction of its agonist. METHODS: There were 20 subjects who were randomized into 3 groups that received either the new hybrid program (HYB), isotonic weight training (WT), or isometric electrical stimulation (ES). Subjects trained 3 times per week for 8 wk with each session consisting of their non-dominant upper extremity performing 10 sets of 10 reciprocal 2-s elbow flexion and extension contractions separated by 1-min rest intervals. Elbow flexion\extension torques and biceps\triceps cross-sectional areas (CSA) were measured at the beginning, midpoint, and end of training as well as at follow-up 4 wk later. RESULTS: The HYB group demonstrated statistically significant increases in elbow flexion and extension torques (56% and 31%, respectively) at the end of training that were similar to or larger than the gains in the other groups with benefits that persisted at follow-up 4 wk later. Muscle CSA increases in the HYB group (10%) were comparable or larger than those in WT and ES subjects. CONCLUSIONS: These results suggest that HYB training may be an effective way to maintain and increase muscle bulk and strength for patients during bed rest, as well as astronauts in space.

Adult↗

[One mechanical property of contraction of isolated bullfrog geniohyoid muscle].

In general, skeletal muscle activity has been understood on the basis that the muscle is functionally consisted of contractile element and elastic element. In this paper it has been attempted to clarify the specific role of the geniohyoid muscle which is one member of the oro-basal muscular system, because our previous reports suggested an elastic specificity of only this muscle as supposed, for example, from the presence of posttetanic twitch attenuation under isotonic condition. For this reason, the experiments have been carried out with paying attention to an interrelationship between loading and contraction figure. When the geniohyoid muscle, weighing over 320 mg, was contracted without any loading, downward undulation under the base line (abbreviated by negative deflection) could be obviously seen within the period succeeding after a falling or relaxation phase of a contraction curve. This negative deflection which presumably indicates the elastic behavior disappeared with increasing the loads from nothing to 4 times the muscle weight. This 3-4 times loading was peculiar to the geniohyoid muscle. In the same situation, 4-5 times loading was specific to the rectus abdominis muscle, and 7-8 times loading was specific to the sartorius muscle. Furthermore, when the geniohyoid muscle was divided first to 1/2 and then to 1/4, the negative deflection of each muscle block was shifted to the lighter side of the applied load. Additional experimental results will be accumulated in the next report.

Animals↗

The contraction-relaxation coupling during pressure-induced cardiac hypertrophy.

Contraction-relaxation coupling was studied in rat and guinea-pig papillary muscles during chronic pressure overload induced by aortic stenosis and during acute hypoxia. Coefficient R1 (ratio between maximum shortening and lengthening velocities of the isotonic twitch loaded with preload only) and coefficient R2 (ratio between the positive and negative peak force derivatives of the isomeric twitch) tested the contraction-relaxation coupling under low and heavy load respectively. Cardiac hypertrophy was similar in guinea-pigs (+43 +/- 5%) and rats (+55 +/- 7%). In both species, cardiac hypertrophy significantly impaired contraction and relaxation phases. In the rat, neither R1 (-1 +/- 4%) nor R2 (-5 +/- 4%) varied significantly during cardiac hypertrophy whereas, in the guinea-pig, an increase in R1 (+56 +/- 18%), P less than 0.001) and in R2 (+26 +/- 9%, P less than 0.01) was noted. These species-related differences might be linked in part to differences in sarcoplasmic reticulum function and myosin ATPase activity. Acute hypoxia, which leads to a decrease in cellular ATP levels, was responsible for a marked decrease in myocardial performance, while R1 increased (+66 +/- 8%, P less than 0.05) and R2 decreased (-14 +/- 1%, P less than 0.05). These results showed that chronic pressure overload modified the contraction-relaxation coupling in a characteristic manner according to the species studied and these changes differed from those observed during acute hypoxia.

Animals↗

Unloaded shortening of skinned mammalian skeletal muscle fibres: effects of the experimental approach and passive force.

In rabbit, rat and human skinned skeletal muscle fibres the length-time relationship of isotonic releases was determined after maximal Ca2+ activation. Slack test experiments provided information about unloaded conditions. Force clamp experiments of different load were extrapolated for zero load and compared with the slack test data. The course length-time relationship for unloaded conditions was similar using both approaches. However, slack test data showed a triphasic shape which could be fitted by three straight lines (phase I, II, III), whereas the data of force clamp experiments exhibited a steady curved shape. Consequently, the instantaneous slopes differed in the two relationships, but the distance which was shortened during the time interval of phase II was similar in both approaches. The ratio between these unloaded shortening velocities resulting from force clamp and slack test experiments was 1.01 +/- 0.05 (SD) (n = 25). The effects of passive force on the velocity of fibre shortening was investigated in skinned rabbit muscle fibres using slack test experiments. A significant increase in the unloaded shortening velocity was observed when the sarcomere length of the fibres was increased to values which exhibited considerable amounts of passive force. The high reproducibility of the isotonic releases required in this study was achieved by improving some methodological details. Using these improved techniques an identity between the relative fibre and sarcomere shortening was observed during the isotonic releases.

Animals↗

Metabolic response to light exercise after exercise-induced rhabdomyolysis.

Inherent compromises in substrate metabolism, or impaired perfusion of muscle may contribute to the occurrence of exercise-induced rhabdomyolysis. In this study, the lactate response of the elbow flexor muscles to light exercise was examined in eight subjects (five males, three females) who previously demonstrated rhabdomyolysis with extreme swelling (ES; n = 4) or no swelling (NS; n = 4) of the upper arm after eccentric exercise. Subjects performed identical light exercise bouts (45 s of rapid isotonic biceps curls consisting of both concentric and eccentric actions at 25% of maximum voluntary contraction force) using their previously eccentrically exercised arm (E-ARM) and control arm, which was not used previously to perform eccentric exercise (C-ARM). Blood lactate concentration ([La]b) was assessed 1.5, 3, 4.5, 6, and 9 min post-exercise. Peak [La]b and the area under the curve (AUC) were compared between the E-ARM of the ES and NS groups and between the C-ARM and E-ARM of the ES group. The AUC did not differ between the E-ARM of the ES and NS groups (P > 0.05) or between the C-ARM and E-ARM of the ES group (P > 0.05). In the ES group, the increase in [La]b after light exercise with the C-ARM [mean (SD) change, delta: 1.98 (0.7) mmol/l] was not different from the increase after exercising the E-ARM [delta: 2.10 (0.7) mmol/l; P>0.05]. Comparing the response of the E-ARM between groups, the increase in [La]b of the NS group [delta: 1.40 (0.4) mmol/l] was not different than that observed in the ES group [delta: 2.10 (0.7) mmol/l; P>0.05). Thus, subjects who had previously exhibited signs of exercise-induced rhabdomyolysis did not show an abnormal response to low-intensity anaerobic exercise.

Adult↗

Measurement of human small airway smooth muscle function in vitro. Comparison of bronchiolar strips and segments.

We have compared isotonic responses to methacholine of human bronchiolar segments and spiral strips. Both types of preparations contracted dose-dependently to methacholine and had a stable intrinsic contractile activity, which was significantly higher in segments (p less than 0.001). ANOVA indicated that the total variation in responses of both strips and segments was similar and was mainly due to a significant between-preparations/within-patients variation. There was a small but significant trend towards a decrease of sensitivity (EC50) in time for both segments and strips. Net contraction, i.e. the difference between resting length and the length at maximal contraction, did not change in time. Limited length-active shortening experiments indicated that 250 mg was a suitable load for both strips and segments. We concluded that, although human bronchiolar strips and segments are functionally comparable, bronchiolar segments are preferable because of their practical and theoretical advantages over bronchiolar strips.

Adult↗

Vagal afferent discharge from gastric mechanoreceptors during contraction and relaxation of the ferret corpus.

Vagal afferent fibres with mechanoreceptive fields in the gastric corpus were studied electrophysiologically in the urethane-anaesthetized ferret in an attempt to assess how changes in muscle tension and length modulate receptor discharge. Single afferent units were spontaneously active and increased their firing on distension of the corpus. During vagally evoked isovolumetric contractions and relaxations, the afferent discharge closely followed changes in intracorpus pressure. Under near-isotonic conditions, changes in both intracorpus pressure and afferent discharge were attenuated to similar degrees despite large changes in corpus volume, although the latter had a modulating influence on afferent discharge. These mechanoreceptors appeared to behave as tension receptors but were also influenced by muscle length as would occur if the receptor were associated with the laminar intramuscular septa of connective tissue which may serve as intramuscular tendons in this tissue.

Action Potentials↗

Action potential in the transverse tubules and its role in the activation of skeletal muscle.

The double sucrose-gap method was applied to single muscle fibers of Xenopus. From the "artificial node" of the fiber, action potentials were recorded under current-clamping condition together with twitches of the node. The action potentials were stored on magnetic tape. The node was then made inexcitable by tetrodotoxin or by a sodium-free solution, and the wave form of the action potential stored on magnetic tape was imposed on the node under voltage-clamp condition (simulated AP). The twitch height caused by the simulated AP's was always smaller than the twitch height produced by the real action potentials, the ratio being about 0.3 at room temperature. The results strongly suggest that the transverse tubular system is excitable and is necessary for the full activation of twitch, and that the action potential of the tubules contributes to about 70 % of the total mechanical output of the normal isotonic twitch at 20 degrees C. Similar results were obtained in the case of tetanic contraction. At a temperature near 10 degrees C, twitches produced by the simulated AP were not very different (85 % of control amplitude) from the twitches caused by real action potentials. This indicates that the excitability of the tubules becomes less necessary for the full activation of twitch as the temperature becomes lower.

Action Potentials↗

Dual mechanism of the relaxing effect of nicorandil by stimulation of cyclic GMP formation and by hyperpolarization.

In addition to previous results from our laboratory showing that nicorandil relaxed vascular smooth muscle by increasing cyclic GMP levels, it was shown to activate K-channels as well, an effect that also leads to relaxation. In the present study, we attempted to differentiate quantitatively between these two effects in isolated bovine coronary artery strips with simultaneous isotonic measurement of length and radioimmunoassay (RIA) determination of cyclic GMP. When the strips were contracted by the thromboxane A2 analogue U 46619 (1 microM) with 10 microM methylene blue added, nicorandil produced 30-50% relaxation without significant changes in cyclic GMP. When in U 46619-contracted strips the hyperpolarizing effect of nicorandil was suppressed by increasing extracellular K+ to 80.4 mM (30-fold), nicorandil caused only 52% relaxation, whereas cyclic GMP increases were not significantly suppressed. Quantitative separation of both mechanisms of relaxation by nicorandil was further achieved through calculation of the cyclic GMP-mediated component from a correlation between increases in cyclic GMP and percentage of relaxation as produced by nicorandil under conditions of inhibited hyperpolarization, i.e., in strips contracted with 1 microM U 46619 or 26.8 mM K+ (10-fold) and exposed to either 30-fold K+ or 10 mM Ba2+. Under both conditions, similar correlations between cyclic GMP and relaxation were obtained. Because U 46619, in addition to its contractile effect, partially antagonized the relaxation by nicorandil without changing cyclic GMP, the correlation was corrected for this effect and indicated a participation of cyclic GMP in the overall relaxant response of approximately 30-40% at low and less than or equal to 80-90% at high concentrations of nicorandil.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

Tension development in highly stretched vertebrate muscle fibres.

1. An apparatus is described by which the length of a selected part of an isolated muscle fibre can be held constant, giving isometric conditions, or alternatively its length can be measured while tension is held constant (isotonic). Control can be switched between length and tension so as to carry out afterloaded contractions with a shortening stop.2. When a part of a fibre with uniform striation spacing is stretched so far that there is presumably no overlap of filaments, the tension developed during an isometric tetanus with this apparatus is very small (not more than 3-5% of the tension developed at optimum length).3. If the tendon ends are held stationary, a fibre with the same initial length develops a large amount of tension (order of 30-40% of tension at optimum length) with a slow time course. This additional tension is due to shortening of the end parts of the fibre, where the striation spacing is smaller and overlap of filaments still exists.4. The resistance to elongation of a part of a fibre where there is no overlap is only slightly increased on stimulation.5. To a first approximation, the results are in good agreement with expectations based on the sliding filament theory. The development of detectable amounts of tension, and of a slight increase of stiffness, on stimulation, are however not expected on the simplest form of this theory; possible explanations are discussed.

Animals↗

Effect of calcium withdrawal on mechanical threshold in skeletal muscle fibres of the frog.

1. Voltage-clamp experiments were performed on frog skeletal muscle fibres using two intracellular micro-electrodes. The threshold for the Na current and the strength-duration curve for mechanical threshold were determined. 2. The change in threshold for the Na current was studied as a function of the external Ca and Mg concentrations which ranged from 0.1 to 50 mM. 3. The resting potential, effective resistance and threshold for the Na current were unchanged when 1.8 mM-Ca was replaced by 3 mM-Mg, indicating that the surface potential and the electrical properties of the fibres were not modified. The additon of 5 mM-EGTA did not affect these parameters. 4. In Ca-free saline (3 mM-Mg and 5 mM-EGTA) the mechanical threshold was significantly increased for short pulses (less than or equal to 20 msec.). In isolated single muscle fibres this effect was observed shortly after applying the Ca-free saline, and was rapidly reversed upon the return to control saline. 5. In isotonic EGTA (85 mM-EGTA) the muscle fibres were depolarized and were unable to contract even if they were hyperpolarized to --90 mV for 12 min prior to stimulation. If 3 mM-Mg was added, most fibres contracted locally. 6. In single muscle fibres caffeine contractures were unmodified after a 30 min exposure to Ca-free saline. 7. It can be concluded that external Ca withdrawal impairs Ca release from the sarcoplasmic reticulum and that external Ca is not essential for triggering contraction.

Animals↗

Long-term effects of altered activity on skeletal muscle.

This paper is a brief summary of some results obtained by stimulating denervated rat soleus (SOL) and extensor digitorum longus (EDL) muscles in vivo with different patterns of stimuli. It is concluded (1) that appropriate stimulation can maintain and restore normal properties in extrajunctional regions of denervated muscle; (2) that SOL and EDL respond differently to identical stimulation, indicating that the two muscles are intrinsically different; (3) that both frequency and amount of stimulation influence contractile properties but to varying degrees depending on the property and on the type of muscle under study; and (4) that the effects of stimulation on isometric and isotonic speed of shortening can be partly accounted for by its effect on excitation-contraction coupling processes and myosin heavy chain (MHC) expression.

Animals↗

Measurement of relaxation in isolated rat ventricular myocardium during hypoxia and reoxygenation.

The effects of hypoxia and subsequent reoxygenation were examined in isolated left ventricular papillary muscles from the rat at 28 degrees C and 33 degrees C. Studies of relaxation were carried out in isometric and physiologically sequenced contractions. In studies of isometric contractions, the following variables were determined: active tension (AT), maximum rate of tension increase (+dT/dt), time to peak tension (TPT), time for tension to fall from peak to 50% of peak tension (RT1/2), maximum rate of tension decline (-dT/dt), isometric peak (-dT/dt/T), and isometric maximum (-dT/dt per T). Variables measured in physiologically sequenced contractions were the slopes of the relation between -dT/dtmax and end systolic length (SIM) and maximum rate of isotonic muscle lengthening (+dL/dtmax) and end systolic length (SIT). Tau, the exponential time constant for isometric relaxation, was also examined. Pronounced changes in active tension and +dT/dt were seen during hypoxia at both temperatures, whereas changes in measured relaxation variables were less prominent and inconsistent. At neither 28 degrees C nor 33 degrees C did TPT or RT1/2 indicate impaired relaxation during hypoxia. Isometric peak -dT/dt declined with hypoxia at both temperatures but the normalised indices, isometric peak -dT/dt/T, peak (-dT/dt per T), and tau showed consistent impairment of relaxation only at 33 degrees C. In physiologically sequenced contractions, SIM suggested impaired relaxation during hypoxia at 28 degrees C but not at 33 degrees C. SIT showed impaired relaxation at both 28 degrees C and 33 degrees C. These findings are consistent with data suggesting impairment of the cardiac relaxing system during hypoxia. Nevertheless, relaxation appears less affected than contraction, and impairment is best seen late in the cardiac cycle.

Animals↗

Differences in load dependence of relaxation between the left and right ventricular myocardium as a function of age in rats.

To determine whether the variation in the magnitude of work load sustained by the left and right ventricles during adulthood and senescence affects the load-dependent aspect of relaxation, posterior papillary muscles from the left and right ventricles of rats at 4, 10, and 20 months of age were studied under variably loaded conditions in vitro. Because of differences between the life spans of Fischer and Sprague-Dawley rats, the functional characteristics of relaxation were investigated to evaluate the possibility of a differential age-associated response in these two strains of animals. The kinetic performance of the diastolic phase of myocardial contraction was measured by assessing the relative time during which load bearing occurred in a series of afterloaded isotonic twitches. This measurement was expressed as the ratio of the duration of afterloaded isotonic shortening and relengthening to the time required for isometric force to decline to the same level during isometric relaxation. A ratio of less than unity identified a load-dependent state whereas a value greater than one reflected a load-independent condition. Results showed that the right myocardium was completely load independent whereas the left myocardium was fully load dependent at all physiological afterloads. Aging reduced the load independence of the right ventricle and the load dependence of the left ventricle in Fischer rats. In contrast, no aging effect on the properties of afterloaded isotonic relaxation was seen in Sprague-Dawley rats. In conclusion, distinct differences exist in the mechanical dynamics of inactivation between the left and right ventricular myocardium. Aging reduced these variations in Fischer rats but had no apparent influence in Sprague-Dawley animals up to 20 months after birth.

Aging↗