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Comparison of contractile state and myosin isozymes of rat right and left ventricular myocardium.

We compared myocardial mechanics and myosin isozymes of right and left ventricular papillary muscles from adult (6 to 8 month old) male rats. Analysis of force velocity relations indicate that right ventricular papillary muscles contract more rapidly than left at light loads (2.68 +/- 0.13 vs 2.18 +/- 0.07 muscle lengths/s measured 75 ms following stimulation, at 0.5 g/mm2; P less than 0.01). Right ventricular papillary muscles had significantly more of the alpha heavy chain containing V1 myosin isozyme and less of the V3 containing beta heavy chain myosin isozyme than left ventricular preparations (P less than 0.05). Papillary muscle and ventricular free wall myosin isozyme distribution were not significantly different within their respective chambers. The presence of a relatively larger proportion of the alpha heavy chain containing myosin isozyme (V1) in right ventricle papillary muscles relative to left correlated with the more rapid velocities of shortening seen in right ventricular papillary muscles (r = 0.60; P less than 0.01).

Animals↗

Effect of age on cholinergic muscarinic responsiveness and receptors in the rat urinary bladder.

The effects of age on urinary bladder responsiveness to muscarinic agonists and on the Bmax and Kd of the binding of [3H]quinuclidinyl benzilate (QNB) to muscarinic receptors of the bladder were studied. Bladder bodies and bases were isolated from Fischer 344 rats, ages seven, 16 and 27 months. No age-dependent change in maximum KCl-elicited isotonic contractions was observed in either bladder region. The bladder base showed an age-dependent increase in the maximum contractions (Emax) elicited by muscarinic agonists. The Emax values for bladder bases from rats 27 months of age were 44 per cent, 58 per cent and 76 per cent greater than those from rats seven months of age for acetylcholine, bethanechol and oxotremorine, respectively. No such alteration in responsiveness was observed in the bladder body with age. There were no age-related changes in ED50 values for the three agonists in either bladder region. Analysis of [3H]QNB binding in the bladder base demonstrated a modest 18 per cent increase in the Bmax (fmol./mg. tissue) from seven to 16 months and a significant 39 per cent decrease from 16 to 27 months. In the bladder body, the Bmax progressively increased by 25 per cent from seven to 27 months. The Kd values of [3H]QNB did not change with age in either region. The data demonstrate that the age-related increase in the responsiveness of the bladder is regionally specific and cannot be explained by a change in the number or affinity of muscarinic receptors.

Acetylcholine↗

A new method for the investigation of the closure function of the resting female urethra.

The field-gradient principle has been used in the development of a probe for the recording of related values of cross-sectional area and pressure in the resting female urethra. The probe is used in a manner that allows pressure to be increased in steps of 10 cm. water, distending the urethra up to a cross-sectional area of 0.79 cm.2. Recordings are performed at 0.5 cm. steps throughout the urethra. The obtained parameters make possible a description of the closure function in terms of urethral rigidity, closure pressure of the noninstrumented urethra, capability of isometric and isotonic contraction, muscular function and power of the closure mechanism.

Female↗

Effects of palmatine on isometric force and intracellular calcium levels of arterial smooth muscle.

The effects of palmatine on isometric force and intracellular free calcium levels ([Ca2+]i) were determined in isolated rat arterial strips. Palmatine dose-dependently relaxed the contractile responses stimulated by phenylephrine (PE) in aortic strips. In contrast, it only partially relaxed aortic strips contracted by 51 mM KCl. Pretreatment with palmatine shifted the dose-response curves of PE both rightwards and downwards in a dose-dependent manner. When Ca2+-free solution and re-addition of Ca2+ were applied to assess PE-induced phasic and tonic contractions, palmatine was found to be effective in inhibiting both contractions. The effects of palmatine on intracellular calcium levels were measured with the bioluminescent calcium indicator aequorin in rat tail artery strips. Palmatine caused a concomitant, dose-dependent decrease in PE-activated isometric force and [Ca2+]i, resulting in small changes in the [Ca2+]i-force relationship. These results suggest that vasodilatory effect of palmatine was mediated by reducing [Ca2+]i as well as affecting [Ca2+]i sensitivity of the contractile apparatus. Palmatine-induced [Ca2+]i decreases appeared to involve decreases in both Ca2+ release from intracellular stores and Ca2+ influx through calcium channels.

Aequorin↗

Potentiation of the effects of bradykinin on its receptor in the isolated guinea pig ileum.

Angiotensin I-converting enzyme (ACE/kininase II) inhibitors potentiated guinea pig ileum's isotonic contractions to bradykinin (BK) and its analogues, shifting the BK dose-response curve to the left. ACE inhibitors added at the peak of the contraction immediately enhanced it further (343 +/- 40%), although the ileum inactivated BK slowly (t(1/2) = 12-16 min). Chymotrypsin and cathepsin G also augmented the activity of BK up to three- or four-fold, but in a manner slower than that of ACE inhibitors. The BK B(2) receptor blocker HOE 140 inhibited all effects. Histamine and angiotensin II were not potentiated. ACE inhibitors potentiate BK independent of blocking its inactivation by inducing crosstalk between ACE and the BK B(2) receptor; proteases activate the receptor by different mechanism.

Animals↗

Improvement of masticatory function after orthodontic treatment. Two case reports.

In patients with temporomandibular joints dysfunction (TMD), the masticatory function is often altered in that: (1) the intercuspal position (ICP) is less frequently achieved; (2) the mean vertical displacement from ICP at the end of opening is above normal; and (3) the isometric muscle contraction pattern during closure tends to be suppressed. We describe two cases of growing patients with severe malocclusion, who showed the same functional alterations as patients with TMJ before treatment, and normalization of the masticatory function after orthodontics.

Child↗

Facilitation of motor evoked potentials after repetitive voluntary hand movements depends on the type of motor activity.

Recent neurophysiological studies suggest that repetitive execution of identical movements is crucial for motor learning. During and after repetitive motor action, changes in motor cortical excitability have been demonstrated by means of transcranial magnetic stimulation. Nevertheless, the frequency and intensity of movement repetition that are necessary to achieve an optimal improvement in motor function are unknown. Fourteen healthy volunteers participated in the present study, which deals with the post-exercise facilitatory and/or inhibitory effects of 5 different motor conditions, including repetitive isotonic contractions at the wrist at two different velocities and two different forearm positions, a sustained isometric hand extension and repetitive hand extensions at the wrist induced by means of transcutaneous electrical muscle stimulation. The modification of muscular response potentials in the extensor carpi radialis muscle was measured following the various motor tasks and the electrical muscle stimulation. The only statistically significant facilitatory effect was observed following an extension-relaxation task at low frequency. Furthermore, the duration of transcranially induced silent periods showed a significant reduction after this motor task.

Adolescent↗

Phenotype modulation in non-adherent and adherent sublines of Walker carcinosarcoma cells: the role of cell-substratum contacts and microtubules in controlling cell shape, locomotion and cytoskeletal structure.

We characterised two sublines of Walker carcinosarcoma cells generated by epigenetic changes. Subline 1 cells were mostly polarised and made no or only non-adhesive cell-substratum contacts. Subline 2 cells were spread, adhesive and mainly non-polar. Subline 1 cells migrate in a non-adhesive mode which is very efficient but operates only in a 3D environment, whereas subline 2 cells migrate in an adhesive mode, which is less efficient but works on 2D and 3D substrata. Nocodazole had little or no effect on shape, polarity and locomotion of subline 1 cells. In glass-adherent subline 2 cells, 10(-6)M nocodazole increased the proportion of polarised cells migrating in an adhesive mode and decreased adhesion to the substratum, whereas 10(-5)M nocodazole further reduced the contacts and the cells reverted to a non-adhesive mode of locomotion. When non-polar subline 2 cells were detached mechanically or by nocodazole, they became polarised and morphologically indistinguishable from non-adherent subline 1 cells. On more adhesive plastic substrata, subline 2 cells produced heterogeneous responses to nocodazole including loss of polarity. The phenotypes of Walker carcinosarcoma sublines have similarities with a broad range of cell types ranging from leucocytes to fibroblast-like cells, suggesting that these phenotypic differences can be controlled by the adhesive and contractile state rather than the cell type. Adhesion modulates contractility (isometric or isotonic contraction) and vice versa and this determines morphology (shape, F-actin, myosin and alpha-actinin), locomotion and responses to microtubule-disassembly. The model may be applied to analyse the mechanisms controlling the phenotype of cells in general.

Actinin↗

Acute and sustained effects of isometric and lengthening muscle contractions on high-energy phosphates and glycogen metabolism in rat tibialis anterior muscle.

Previous studies have shown that lengthening contractions, in contrast to isometric contractions, readily result in sustained malfunctioning of the exercised muscles. The present study was performed to investigate whether an exercise period with many (240) lengthening contractions (LC) results in alterations in muscle high-energy phosphates and inosine monophosphate (IMP) content, different from muscles performing a few (60) lengthening or a few (60) or many (240) isometric contractions (IC). Moreover, we sought for a possible cause(s) of the inability to replenish muscle glycogen stores following LC. Rat tibialis anterior muscles were subjected in vivo to either 60 or 240 LC or IC. Structural muscle damage occurred only after 240 LC. The fact that tissue glycogen levels declined to a similar extent during LC and IC suggests that the energy demand was comparable during both types of exercise. Nevertheless, the observation that on the one hand tissue stores of adenine nucleotides showed a greater decline, and on the other hand the tissue content of IMP increased to a significantly higher level after LC than after IC, clearly indicates that muscle energy metabolism is more disturbed during LC than during IC. The high tissue levels of IMP may contribute to impaired mechanical function as previously observed in muscles subjected to LC. In contrast to 240 IC, 24 hours after 240 LC, tissue glycogen stores and high-energy phosphate levels were not restored to control values. The present findings indicate that depressed glycogen synthase activity and impaired activity of the mitochondrial marker enzyme cytochrome C oxidase probably contribute to a continuous disturbance of energy metabolism in the exercised muscles during the 24 hours following 240 LC.

Adenosine Diphosphate↗

Crossbridge kinetics in single frog muscle fibres in presence of ethylene glycol.

Single fibres isolated from frog muscle were tetanically stimulated at 14 degrees C to produce isometric tetani at a sarcomere length of about 2.16 microm, using a striation follower device to measure the sarcomere length of a selected segment of fibre. Force-velocity data were obtained by applying ramp releases at pre-set velocity at the tetanus plateau. Sarcomere stiffness was measured at isometric plateau and during isotonic shortening by using sinusoidal length changes at 2 kHz frequency and about 1 nm per half sarcomere (hs) peak to peak amplitude. A correction method was used to compensate for the force truncation due to the quick recovery. After data collection, the bathing solution was substituted with Ringer plus ethylene glycol (EG) at 2 M (11.2% v/v). When the fibre was fully equilibrated with the new solution, the measurements were repeated. Ethylene glycol reduced the speed of the tetanus rise and tetanus relaxation without altering the isometric tension, and reduced the maximum shortening velocity by about 20%. During isotonic contraction tension and stiffness at each given shortening velocity were reduced by about the same amount, so that the stiffness/tension ratio remained almost unaltered. Force-velocity and stiffness data in both standard and EG Ringer were analysed in terms of a two state model (Huxley, 1957). The analysis showed that our results can be accounted for by assuming that EG at 2 M concentration reduces all the rate constants for crossbridges interaction by about the same amount.

Animals↗

5-HT(4) receptors on cholinergic nerves involved in contractility of canine and human large intestine longitudinal muscle.

5-HT(4) receptors mediate circular muscle relaxation in both human and canine large intestine, but this phenomenon alone can not explain the improvement in colonic motility induced by selective 5-HT(4) receptor agonists in vivo. We set out to characterize 5-HT(4) receptor-mediated effects in longitudinal muscle strips of canine and human large intestine. Electrical field stimulation (EFS) was applied providing submaximal isotonic contractions. L-NOARG (0.1 mM) was continuously present in the organ bath to preclude nitric oxide-induced relaxation to EFS. The selective 5-HT(4) receptor agonist prucalopride (0.3 microM) enhanced EFS-evoked contractions, that were antagonized in both preparations by the selective 5-HT(4) receptor antagonist GR 113808 (0.1 microM). The prucalopride-induced increase was present in canine ascending and descending colon, but absent in rectum. Regional differences in response to prucalopride were not observed in human ascending and sigmoid colon and rectum. Incubation with atropine (1 microM) or tetrodotoxin (0.3 microM) inhibited EFS-induced contractions, which were then unaffected by prucalopride (0.3 microM) in both tissues. In the presence of methysergide (3 microM; both tissues) and granisetron (0.3 microM; only human tissues), 5-HT (0.3 microM) enhanced EFS-induced contractions, an effect that was antagonized by GR 113808 (0.1 microM). In the presence of atropine or tetrodotoxin, EFS-induced contractions were inhibited, leaving 5-HT (0.3 microM) ineffective in both preparations. This study demonstrates for the first time that in human and canine large intestine, 5-HT(4) receptors are located on cholinergic neurones, presumably mediating facilitating release of acetylcholine, resulting in enhanced longitudinal muscle contractility. This study and previous circular muscle strip studies suggest that 5-HT(4) receptor agonism facilitates colonic propulsion via a coordinated combination of inhibition of circumferential resistance and enhancement of longitudinal muscle contractility.

Animals↗

A physiotherapeutic approach to craniomandibular disorders: a case report.

This is a case report of a 19-year-old female who presented with a unilateral weakness of the right masseter muscle evidenced by electromyographic examination. The presence of a mandibular deviation to the right during opening because of this weakness was treated with neuromuscular electrical stimulation (NMES). After the physiotherapeutic treatment, the electrical activity of the right masseter muscle increased during function and the mandibular deviation disappeared. Electromyography (EMG) can have a useful role in the determination of the muscular profile, and for evaluating therapeutics.

Adult↗

Contractile effects and intracellular Ca2+ signalling induced by motilin and erythromycin in the circular smooth muscle of human colon.

Motilin has excitatory effects on the colon of the rabbit and the dog, but little is known of its effect on the human colon. The aim of this study was to investigate the effects induced by motilin and erythromycin A (EMA) on muscle strips and on single cells from primary cultures from human colon. Isotonic contraction was recorded in circular muscle strips from macroscopically normal resection specimens of patients operated on for colonic neoplasm. Agonist-induced intracellular Ca2+ ([Ca2+]i) signalling was studied in primary cultures of colonic smooth-muscle cells using the ratiometric Ca2+ indicator Indo 1, on a laser-scanning confocal epifluorescence microscope. In circular muscle strips, norleucine13-porcine motilin ([Nle13]-pm)and EMA induced tonic contractions with an EC50 of 92 +/- 21 nmol L(-1) and 31 +/- 16 micromol L(-1), respectively. The maximal contraction was 21 +/- 4% (motilin) and 33 +/- 12% (EMA) of the response to 10(-4) mol L(-1) acetylcholine (ACh). The motilin antagonist OHM-11526 (10(-5.5) mol L(-1)) abolished the effects of both [Nle13]-pm and EMA. Neither tetrodotoxin (10(-5.5) mol L(-1)), L-nitro-D-arginine methyl ester (L-NAME) (10(-3.5) mol L(-1)) nor guanethidine (10(-5) mol L(-1)) interfered with the effects of [Nle13]-pm or EMA. [Nle13]-pm (10(-11)-10(-6) mol L(-1)) induced rises of [Ca2+]i in cultured colonic myocytes. At 10(-6) mol L-1, 94% of the cells responded, and half of the cells responded at 1.4 nmol L(-1) [Nle13]-pm. 81% (35/43) and 95% (75/79) responded to EMA (10(-6) mol L(-1)) and acetylcholine (ACh, 10(-4) mol L(-1)), respectively. The motilin antagonist GM-109 inhibited motilin- and EMA-induced [Ca2+]i rises. In the absence of extracellular Ca2+, only 13% (7/52) of the cells responded to [Nle13]-pm (10(-6) mol L(-1)) vs. 90% (47/52) to ACh (10(-4) mol L(-1)). Motilin and EMA have direct excitatory effects on circular smooth muscle from the human colon and these effects are mediated via a smooth-muscle motilin receptor. These findings suggest that motilin may regulate colonic motility and that motilides may have therapeutic potential for the treatment of colonic hypomotility.

Acetylcholine↗

Increased resistance against shortening in myocardium from recipient hearts of 7 patients transplanted for dilated cardiomyopathy.

UNLABELLED: The contractile behaviour of demembranized atrial and ventricular myocardium of 7 patients transplanted for end-stage heart failure (ESHF) was analyzed. Atrial muscle specimens of patients undergoing coronary artery bypass surgery (n = 9) and pig papillary muscle were used as reference preparations (n = 9). Extreme care was taken for dissection and mounting the muscle fibres (0.3 x 6 mm) in order to keep the passive series compliance small. Calcium sensitivity, cross-bridge cycling rate (estimated by the force-clamping technique and calculation of the shortening velocity at zero load [Vmax]) and isometric force development were measured. Analysis on light- and electronmicroscopic level was carried out. RESULTS: 1) Calcium sensitivity was not altered in ESHF patients; 2) the velocity of the force generating process (cross-bridge cycling rate) was normal in ventricular and reduced in atrial ESHF myocardium, 3) maximum isometric force development was reduced in ventricular, but not in atrial myocardium of ESHF patients, and 4) Vmax was significantly reduced in ventricular and atrial ESHF myocardium (p < 0.0001). Perimysial and endomysial fibrosis was present in ventricular, not in atrial myocardium of ESHF patients. CONCLUSION: A normal cross-bridge cycling rate in left-ventricular ESHF myocardium combined with a decreased capability of muscle shortening indicates the presence of a resistance against shortening localized either on the cross-bridge level or/and due to intra- and pericellular fibrosis. Left-ventricular contractile dysfunction in patients with end-stage heart failure may be related to a normal contractile apparatus contracting within an abnormal intracellular or interstitial environment.

Animals↗

Contractile behaviour of skinned papillary muscle in mitral valve disease.

The contractile behaviour of Triton-X 100 skinned left ventricular papillary muscle from 19 patients undergoing cardiac surgery for mitral valve stenosis: n = 6, mitral valve incompetence: n = 7, or combined mitral valve disease: n = 6 was analyzed. At supramaximal activation the "vibration induced force clamping technique" was used for isometric analysis of time course and extent of isometric postvibration force recovery. Afterloaded contractions were applied for extrapolation of the maximum shortening velocity at zero load (Vmax). The Calcium sensitivity was analysed by variation of the free EGTA-buffered Calcium concentration at a passive resting force of 2 mN at 26 degrees C. In different types of mitral valve disease the characteristics of isometric force development were unaltered in terms of maximum force development, force per square mm, Calcium sensitivity and the time course of isometric contraction after force clamping. However the capability to shorten as expressed by Vmax was reduced in mitral valve incompetence (3.87 +/- 0.37 ML/s) as compared with mitral valve stenosis (5.29 +/- 0.35 ML/s) or combined mitral valve disease (4.83 +/- 0.51 ML/s). The ratio between the inverse value of Vmax and the time constant of isometric force development after force clamping was significantly different in mitral valve incompetence as compared with other types of mitral valve disease (p < 0.0001). These data argue for the presence of different resistances against shortening in various types of mitral valve disease, due to altered cross-bridge cycling characteristics or to morphological factors.

Adult↗

Force development of dynamic and static contractions in children and adults.

There are still unclarified points remaining in regard to the characteristics of force development in children who are immature in their basic activities in daily living. Moreover, there are morphological, histological, and biochemical differences between immature and mature muscles. In the present study, dynamic elbow flexion and static contractions which were performed by elbow flexors with maximum effort were examined by force-velocity and force-time relationships and electromyograms (EMG) in six-year-old children and in adults. In dynamic contractions with various loads, the force-velocity relationship for children was (P + 5.65) (v + 1.48) = 19.51, and (P + 14.65) (v + 2.26) = 96.21 for adults. The maximum power in children and adults was 22.8 watts and 160.8 watts, respectively. The contraction delay in children was greater than that in adults for every relative load. In static contractions, the maximum rate of force development for children was significantly lower than that for adults at each pre-tension. With an increase in pre-tension, the adult rate showed a tendency to decrease, whereas the rate for children showed no change. The electromechanical delay observed in children was significantly greater than that in adults. For dynamic and static contractions, the contractile speed in children was lower than that in adults.

Adult↗

Damped elastic recoil of the titin spring in myofibrils of human myocardium.

The giant protein titin functions as a molecular spring in muscle and is responsible for most of the passive tension of myocardium. Because the titin spring is extended during diastolic stretch, it will recoil elastically during systole and potentially may influence the overall shortening behavior of cardiac muscle. Here, titin elastic recoil was quantified in single human heart myofibrils by using a high-speed charge-coupled device-line camera and a nanonewtonrange force sensor. Application of a slack-test protocol revealed that the passive shortening velocity (Vp) of nonactivated cardiomyofibrils depends on: (i) initial sarcomere length, (ii) release-step amplitude, and (iii) temperature. Selective digestion of titin, with low doses of trypsin, decelerated myofibrillar passive recoil and eventually stopped it. Selective extraction of actin filaments with a Ca2+-independent gelsolin fragment greatly reduced the dependency of Vp on release-step size and temperature. These results are explained by the presence of viscous forces opposing myofibrillar passive recoil that are caused mainly by weak actin-titin interactions. Thus, Vp is determined by two distinct factors: titin elastic recoil and internal viscous drag forces. The recoil could be modeled as that of a damped entropic spring consisting of independent worm-like chains. The functional importance of myofibrillar elastic recoil was addressed by comparing instantaneous Vp to unloaded shortening velocity, which was measured in demembranated, fully Ca2+-activated, human cardiac fibers. Titin-driven passive recoil was much faster than active unloaded shortening velocity in early phases of isotonic contraction. Damped myofibrillar elastic recoil could help accelerate active contraction speed of human myocardium during early systolic shortening.

Actins↗