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Embryonic chicken skeletal, cardiac, and smooth muscles express a common embryo-specific myosin light chain.

It has been demonstrated that embryonic chicken gizzard smooth muscle contains a unique embryonic myosin light chain of 23,000 mol wt, called L23 (Katoh, N., and S. Kubo, 1978, Biochem. Biophys. Acta, 535:401-411; Takano-Ohmuro, H., T. Obinata, T. Mikawa, and T. Masaki, 1983, J. Biochem. (Tokyo), 93:903-908). When we examined myosins in developing chicken ventricular and pectoralis muscles by two-dimensional gel electrophoresis, the myosin light chain (Le) that completely comigrates with L23 was detected in both striated muscles at early developmental stages. Two monoclonal antibodies, MT-53f and MT-185d, were applied to characterize the embryonic light chain Le of striated muscles. Both monoclonal antibodies were raised to fast skeletal muscle myosin light chains; the former antibody is specific to fast muscle myosin light chains 1 and 3, whereas the latter recognizes not only fast muscle myosin light chains but also the embryonic smooth muscle light chain L23. The immunoblots combined with both one- and two-dimensional gel electrophoresis showed that Le reacts with MT-185d but not with MT-53f. These results strongly indicate that Le is identical to L23 and that embryonic chicken skeletal, cardiac, and smooth muscles express a common embryo-specific myosin light chain.

Animals↗

Acute heat stress stimulates mitochondrial superoxide production in broiler skeletal muscle, possibly via downregulation of uncoupling protein content.

Acute heat stress (34 degrees C for 18 h) resulted in increased levels of reactive oxygen species (ROS) in mitochondria isolated from the skeletal muscle of broilers. This occurred when glutamate-requiring complexes I, III, and IV of the electron transport chain or succinate-requiring complexes II, III and IV were used as the substrate. This result confirms our previous observation that exposure of broilers to 34 degrees C for 18 h results in increased superoxide production in skeletal (pectoralis) muscle, and extends this finding by showing that substrate-independent ROS generation occurs during the heat stress period. When broilers were exposed to heat stress, the levels of avian uncoupling protein (avUCP) mRNA in skeletal muscle were significantly decreased, to 28% of the levels found in untreated controls. This was accompanied by a significant decrease in the levels of the avUCP protein, to 37% of control levels. In contrast, avian adenine nucleotide translocator mRNA levels were not affected by exposure to heat stress. This finding is consistent with previous studies which showed that the increases in superoxide production that are observed in the presence of carboxyatractylate, a specific inhibitor of adenine nucleotide translocator, were the same for skeletal muscle mitochondria from both control and heat-stressed chickens. Taken together, these results suggest that acute heat stress stimulates mitochondrial superoxide production in broiler skeletal muscle, possibly via downregulation of avUCP. The present study provides the first evidence that synthesis of avUCP protein is downregulated in heat-stressed broilers.

Animals↗

Contractile properties and protein isoforms of single fibres from the chicken pectoralis red strip muscle.

1. The contractile properties of single muscle fibres of the red strip region of adult chicken pectoralis major (PM) muscle, some of which are known to express an embryonic isoform of myosin heavy chain (MHC), were determined and compared with the properties of the fast white fibres of the PM and the slow tonic fibres of the anterior latissimus dorsi (ALD) muscle. 2. The red strip fibres could be classified into two groups, fast and slow. The mean velocity of unloaded shortening (Vmax) in fast red strip fibres was approximately half the Vmax of fast white fibres. Vmax of slow red strip fibres was less than 20% of the value for fast red strip fibres and was not different from Vmax of ALD fibres. 3. The tension-generating ability, i.e. the maximal isometric tension/fibre cross-sectional area (P0/CSA), was the same in fast red strip fibres and fast white fibres. P0/CSA was approximately 30% lower in slow red strip fibres compared with fast red strip fibres but was 70% greater in slow red strip fibres compared with ALD fibres. 4. The tension-pCa relation of fast red strip fibres was shifted to lower pCa values, indicating a lower calcium sensitivity compared with fast white fibres, and this difference was associated with a difference in troponin T isoform composition. The tension-pCa relation of slow red strip fibres was not different from that in ALD fibres. 5. The difference in Vmax between fast red strip fibres and fast white fibres was associated with different MHC compositions of these fibres. 6. The myofibrillar protein isoform composition of slow red strip fibres was identical to that of the slow tonic fibres of ALD muscle and these two groups of fibres had very similar contractile properties.

Animals↗

Adenylate cyclase in muscular dystrophy.

The purpose of this study was to determine whether the previously reported differences in adenylate cyclase activity between the sarcolemma of normal and dystrophic chick muscles are also found in the SR, to search for a possible relationship between the adenylate cyclase changes and the pathophysiology of dystrophy, and to investigate whether the findings can be extended to Duchenne human muscular dystrophy by studying the adenylate cyclase and ATPase activities of erythrocyte ghosts from DMD patients and carriers. Microsomes were separated by standard techniques from the pectoralis muscles of normal and dystrophic ckeckens of various ages. The microsomal yields were significantly larger in dystrophic muscles. Adenylate cyclase activities in dystrophic microsomes were higher than those in matched controls and increased with the progression of the disease. The ratio between the two rose from one at 2 weeks of age to nine at about 9--10 weeks. Kinetic analyses showed that the ks for MgATP2- was about 40 microM (at 3 mM Mg2+ and 0.3 mM Ca2+) both in normal and dystrophic microsomes, that calcium caused umcompetitive inhibition of the enzyme (Ki = 0.2 mM), that the effect of calcium was noncooperative (Hill coefficient, nH = 1), that calcium did not affect the cooperativity for MgATP2-, and that magnesium competitively removed the calcium inhibition and caused additional, cooperative stimulation of the enzymatic activity (ka = 1.5 mM; NH =2). The major difference between normal and dystrophic adenylate cyclase was a higher enzymatic velocity in the latter, suggesting a larger amount of enzyme. We investigated whether altered cAMP levels may effect calcium accumulation. Calcium uptake measured (in the presence of oxalate) at several ages revealed no difference between normal and dystrophic chickens. The extent of calcium binding was also similar, although the kd for Ca2+ was lower in dystrophic microsomes. Binding was enhanced in the presence of exogenous protein kinase, but the responses of normal and dystrophic tissues were similar. We concluded that the elevation of adenylate cyclase in dystrophy was not related to microsomal calcium accumultion. Ivestigation of the localization of microsomal adenylate cyclase supported this view. Separation of calcium-loaded microsomes on a discontinuous sucrose gradient into four fractions demonstrated that adenylate cyclase activity, measured in the presence of Lubrol-PX and EGTA, was inversely related to calcium-accumulating activity. Na+, K+-ATPase comigrated with adenylate cyclase. Highest specific activities were found in the lightest fraction. These observations were confirmed by histochemical studies. The reaction product from adenylate cyclase activity was present predominantly in the terminal cisternae of the SR. In the context of the literature, our findings suggest that the rises in adenylate cyclase and Na+, K+-ATPase in avian dystrophy are compensatory changes, elicited by a defect in ECC at the calcium release step...

Adenylyl Cyclases↗

The mechanical power output of the flight muscles of blue-breasted quail (Coturnix chinensis) during take-off.

Blue-breasted quail (Coturnix chinensis) were filmed during take-off flights. By tracking the position of the centre of mass of the bird in three dimensions, we were able to calculate the power required to increase the potential and kinetic energy. In addition, high-speed video recordings of the position of the wings over the course of the wing stroke, and morphological measurements, allowed us to calculate the aerodynamic and inertial power requirements. The total power output required from the pectoralis muscle was, on average, 390 W kg(-1), which was similar to the highest measurements made on bundles of muscle fibres in vitro (433 W kg(-1)), although for one individual a power output of 530 W kg(-1) was calculated. The majority of the power was required to increase the potential energy of the body. The power output of these muscles is the highest yet found for any muscle in repetitive contractions. We also calculated the power requirements during take-off flights in four other species in the family Phasianidae. Power output was found to be independent of body mass in this family. However, the precise scaling of burst power output within this group must await a better assessment of whether similar levels of performance were measured across the group. We extended our analysis to one species of hawk, several species of hummingbird and two species of bee. Remarkably, we concluded that, over a broad range of body size (0.0002-5 kg) and contractile frequency (5-186 Hz), the myofibrillar power output of flight muscles during short maximal bursts is very high (360-460 W kg(-1)) and shows very little scaling with body mass. The approximate constancy of power output means that the work output varies inversely with wingbeat frequency and reaches values of approximately 30-60 J kg(-1) in the largest species.

Animals↗

Effect of Bothrops insularis venom on the mouse and chick nerve-muscle preparation.

The effects of Bothrops insularis venom were examined in vivo in mice and chicks and in vitro using the mouse phrenic nerve diaphragm and chick biventer cervicis muscle preparations. Incubation of the indirectly or directly stimulated mouse preparation with B. insularis venom (20-80 micrograms/ml) produced an initial increase in twitch tension followed by irreversible blockade. With direct stimulation in the presence of D-tubocurarine, no increase in twitch tension was observed prior to the onset of blockade. A venom-induced effect on presynaptic activity was suggested by the marked increase in the frequency of the mepps recorded in vitro 5-15 min after venom addition. A direct muscular effect was shown by the dose- and time-dependent reduction in the resting membrane potential of the diaphragm. Chick preparations were more sensitive than those of the mouse. In the isolated chick biventer cervicis muscle preparation, B. insularis venom induced a contracture and a dose-dependent block of responses to indirect stimulation. At low venom concentrations (1-5 micrograms/ml), no significant release of creatine kinase (CK) was observed from this preparation. However, a dose-dependent release of CK was detected at higher doses (10-80 micrograms/ml). For morphological studies, B. insularis venom was injected into the chick left pectoralis muscle. At low doses (0.4 microgram), only an inflammatory reaction was present, while at high doses (20-80 micrograms) increasing numbers of necrotic fibers were observed as well as occlusive thrombosis and hemorrhage. The muscular effect, also observed on the incubated muscle, points to a direct myolytic action of the whole venom.

Animals↗

Metabolic and myogenic factors in local regulation of the microcirculation.

Patterns of flow were recorded from individual capillaries of mesentery and muscle during autoregulation and reactive hyperemia. In cat mesentery at normal arterial pressure capillary blood flow was often periodic in nature. When arterial pressure was reduced periodicity was abolished and in certain cases mean flow increased. Elevation of venous pressure at this time caused restoration of flow periodicity and simultaneously a large fall in mean flow. Vasomotion and autoregulation in mesentery appear to be dependent on intravascular pressure per se. In cat sartorius muscle substantial increase in flow was seen in most capillaries during reactive hyperemia. In certain capillaries the pattern resembled the gross flow pattern while others showed a brief hyperemia and then a period of flow arrest that is presumably due to a strong precapillary vasoconstriction. The latter response is suggestive of a myogenic control while the former may be due to accumulation of metabolites. In frog pectoralis muscle reactive hyperemia was very prolonged in comparison to cat sartorius muscle. The general pattern of flow was consistent with the notion of a strong metabolic control mechanism. The three tissues studied provide examples of strong myogenic, strong metabolic, and combined metabolic and myogenic control of the microcirculation.

Animals↗

The combined use of the TRAM and expanders/implants in breast reconstruction.

The transverse rectus abdominis musculocutaneous (TRAM) flap has replaced the prosthetic implant as the first choice for breast reconstruction. However, we have found several incidences when the volume of autogenous tissue cannot fulfill the requirements for symmetry, especially in those patients with limited abdominal tissue and large ptotic breasts. The TRAM flap can be combined with tissue expanders and implants to obtain symmetry in these difficult reconstructive situations. Here we present our experience with the use of the TRAM flap in conjunction with expanders and saline implants. Between June 1993 and November 1996, 4 patients underwent immediate TRAM reconstruction in conjunction with saline implants or tissue expanders. In 3 patients a tissue expander was placed beneath the pectoralis muscle following the microanastomoses and prior to insetting the flap. Expansion was initiated 2 to 6 weeks postoperatively and continued over a period of 1 to 3 months. Permanent implants were placed at 3 to 4 months following the initial procedure. The final patient underwent a bilateral free TRAM with the placement of bilateral saline implants at the time of the initial reconstruction. One implant was placed beneath the pectoralis whereas the other was placed beneath the TRAM flap. Saline implants were used in all patients. The decision to use the combination of implants and TRAM flap was multifactorial and included position of donor scar in all patients, large ptotic breasts in 2 patients, request for contralateral augmentation in 2 patients, and an absence of adequate abdominal tissue in all patients. In addition, large skin deficits were present in 3 patients following mastectomy requiring the skin coverage provided by the TRAM flap. The average age of the patients was 41 years. Expanders were left in place for an average of 16 weeks. Average final implant volume was 210 cc. The average follow-up was 24 months, during which time there were no episodes of flap compromise, partial flap loss, or fat necrosis. The TRAM flap in combination with tissue expansion and implants provides an additional alternative in postmastectomy reconstruction. This technique is particularly useful in those patients in whom the need for TRAM skin coverage is combined with inadequate autogenous tissue and the patient request for large ptotic breasts.

Adult↗

Primary structure of chicken skeletal muscle and fibroblast alpha-actinins deduced from cDNA sequences.

The complete 897-amino-acid sequence of chicken skeletal muscle alpha-actinin and the 856-amino-acid sequence (97% of the entire sequence) of chicken fibroblast alpha-actinin have been determined by cloning and sequencing the cDNAs. Genomic Southern analysis with the cDNA sequences shows that skeletal and fibroblast alpha-actinins are encoded by separate single-copy genes. RNA blot analyzes show that the skeletal alpha-actinin gene is expressed in the pectoralis muscle and that the fibroblast gene is expressed in the gizzard smooth muscle as well as in the fibroblast. The deduced skeletal alpha-actinin molecule has a calculated Mr of 104 x 10(3), and each alpha-actinin can be divided into three domains: (1) the NH2-terminal highly conserved actin-binding domain, which shows similarity to the product of the Duchenne's muscular dystrophy locus; (2) the middle rod-shaped dimer-forming domain, which contains the spectrin-type repeat units; and (3) the COOH-terminal two EF-hand consensus regions. Comparison of the skeletal alpha-actinin sequence with the fibroblast and smooth muscle alpha-actinin sequences demonstrated that the EF-hand structure was conserved in all of these alpha-actinin sequences, despite the reported variability of the Ca2+ sensitivities of the actin-gelation by various alpha-actinin isoforms.

Actinin↗

Pectoralis major myocutaneous flap reconstruction of the laryngopharynx and cervical esophagus.

Advanced carcinoma of the hypopharynx and cervical esophagus is a formidable challenge to the skills of the head and neck surgeon. Radiation therapy is valuable as adjunctive therapy when combined with curative surgery, which is the primary treatment modality. The extent of anatomical disease associated with extensive neoplasia of the hypopharynx and cervical esophagus is frequently not amenable to total laryngectomy with local tissue repair. Surgical ablation usually requires an extended laryngectomy, which does not permit primary local repair. A previous report by this author, comparing all techniques historically and chronologically, indicated that the present impetus is toward procedures characterized by a one-stage primary repair with shorter completion times. Presently, the three most promising procedures that meet these criteria are the gastric transposition, free microvascular bowel transfer, and regional myocutaneous flap repair. Theogaraj, et al. reported the use of a partially tubulated pectoralis muscle flap over preserved posterior wall cervical esophageal mucosa in cases of short segment stenosis. Encouraged by these results, a technique using partial tubulation for long-segment stenosis was reported. The use of this technique was expanded to include the repair of the defect left after total ablation of the laryngopharynx and cervical esophagus. Over the past 40 months, 22 patients have undergone repair using partial tubulation of the pectoralis myocutaneous flap. This paper will discuss the technical aspects of the procedure and analyze the procedure as it relates to mortality, morbidity, and completion time. Low morbidity and a completion time of 18 days competes favorably with gastric transposition and free jejunal transfer. A rational approach to reconstruction using all procedures will be discussed.

Aged↗

The use of halothane and succinylcholine to identify broilers prone to developing pale, soft, exudative meat.

Within the last several years, the poultry industry has seen a dramatic increase in the occurrence of pale, soft, and exudative (PSE) meat. This problem is known to be associated with a rapid decline in postmortem (PM) muscle pH, which results in inferior protein functionality similar to that found in PSE pork. Many factors such as seasonal changes have been known to influence the occurrence of PSE meat in poultry and swine. Halothane and succinylcholine have been used within the pork industry to identify animals susceptible to stress and prone to developing PSE meat. The mechanism for the triggering of the PSE gene in poultry has not been fully understood. Therefore, a study was conducted to determine the effectiveness of screening broilers with halothane to identify those prone to developing PSE meat. Succinylcholine was used before slaughter to serve as a triggering agent for the PSE condition. At 4 wk of age, broilers from 4 commercial strains (n = 1,000) were subjected to 3% halothane gas and classified as either halothane positive (HAL+) or negative (HAL-) based on muscle rigidity within the legs. Although halothane sensitivity varied slightly among the strains, approximately 14% of the birds overall were classified as HAL+. All HAL- birds (n = 163) and an equal number of HAL-birds (n = 163) in each strain were grown to market age (7 wk) and were commercially processed. At the time of processing, half of the HAL+ and HAL- birds were injected intravenously with succinylcholine and were slaughtered at 0.25 h postinjection. Pectoralis muscle samples were collected at 0.25, 2, 5, and 24 h PM for the evaluation of rigor development (muscle pH) and meat quality (L* value, moisture, drip loss, and cook loss). Halothane sensitivity had no effect on rigor development, muscle color, or water-holding capacity in the 4 broiler strains. Although birds exhibited reactions to the halothane gas, the halothane sensitivity, along with the use of succinylcholine, was not able to identify birds prone to developing PSE meat.

Animals↗

Methionine deficiency decreases protein accretion and synthesis but not tRNA acylation in muscles of chicks.

The effect of supplementing a methionine-deficient, isolated soy protein diet (0.5% total sulfur amino acids) with 0.2% D,L-methionine (DL-MET) or a molar equivalent of D,L-2-hydroxy-4-(methylthio) butanoic acid (DL-HMB) was assessed in chicks over an 8-d feeding study. Chicks consumed DL-HMB diet ad libitum (HIGH) or were restricted to the level consumed ad libitum by chicks fed the basal diet. The DL-MET diet was fed at the same two levels as the DL-HMB diet. Supplementing with either methionine source resulted in significantly greater growth rate, efficiency of feed conversion, and accretion and synthesis of protein in the gastrocnemius and pectoralis muscles. These increases were greater for chicks consuming feed ad libitum as compared with feed-restricted chicks and were not affected by methionine source. Rate of muscle protein degradation appeared to increase with supplementation of either methionine source, but only when feed intake was permitted to increase. The relative in vivo conversion of DL-HMB vs. D-MET to L-MET was similar in all groups as indicated by pool sizes of methionine, tRNA(met), and tRNA(cys), and rates of protein accretion and synthesis. These data demonstrate that dietary DL-HMB and DL-MET are used with similar efficacy to support skeletal muscle protein accretion and rates of protein synthesis when feed intake is equalized.

Acylation↗

Influence of physical exercise after long-lasting hypodynamia on the morphological parameters of muscle fibers.

The purpose of this research was morphometric ultrastructure evaluation of the fibers in muscles taking part in the flight of pigeons forced to exercise after a long period of hypodynamia. It was found that following physical exercise, after 12 and 18 months of mobility limitation, there appeared marked qualitative and quantitative changes: a diminution of the volume fraction and number of mitochondria, increase of smooth sarcoplasmic reticulum and sarcoplasm, and a significant decrease of the number of glycogen granules as compared with those after 18 months hypodynamia. The above described changes were more pronounced in the supracoracoideus than in the pectoralis muscle.

Animals↗

Artificial neural network model of the mapping between electromyographic activation and trajectory patterns in free-arm movements.

Artificial neural networks (ANNs) have been used to identify the relationship between electromyographic (EMG) activity and arm kinematics during the execution of motor tasks. Although considerable work has been devoted to showing that ANNs perform this mapping, there has been little work to explore any relationship with physiological properties of the neuromuscular systems. A back-propagation through time (BPTT) ANN was used to map the EMG of five selected muscles (pectoralis major (PM), anterior deltoid (AD), posterior deltoid (PD), biceps brachii (BB) and triceps brachii (TB)) on arm kinematics in seven normal subjects performing three-dimensional unrestrained grasping movements. To investigate the physiological validity of the BPTT-ANN, inputs were artificially altered, and the predicted outputs were analysed. Results show that the BPTT-ANN performed the mapping correctly (root mean square (RMS) error between target and predicted outputs averaged across subject test sets was 0.092 +/- 0.015). Moreover, it provided insights into the roles of muscles in performing the movement (average indexes measuring the output alteration with respect to the target were 0.070 +/- 0.027, 0.356 +/- 0.172, 0.568 +/- 0.413, 0.510 +/- 0.268, 0.681 +/- 0.430 for PM, AD, PD, BB, TB, respectively, in the movement forward phase, and 0.077 +/- 0.015, 0.179 +/- 0.147, 0.291 +/- 0.247, 0.671 +/- 0.054, 0.232 +/- 0.097 in the return phase).

Adult↗

Naturally-occurring degeneration in chick muscle development: ultrastructure of the M. complexus.

Ultrastructural study of the M. complexus muscle of the chick embryo has demonstrated two populations of fibres: the more common is the normal myotube or muscle fibre, which was observed in various stages of development; the other shows myofibrillar contractions which, at their greatest degree, produce the appearance of an amorphous mass of myofilaments. The contracted fibre had rounded and swollen mitochondria, and vacuoles (autophagic) containing glycogen, and it exhibited a cleavage of the cell which isolated the nuclear region from the main body of the fibre. When the fibres were less contracted they resembled degenerative mammalian fast-twitch-oxidative-glycolytic fibres after immobilization. The more normal fibre population was identical with that of the pectoralis muscle, which was used as a control. These results suggest that the contracted fibres are degenerating, which agrees with conclusions by earlier investigators using light miscroscopy.

Animals↗

[Augmentation mammaplasty through the external oblique muscle route].

To reduce the rate of capsule contracture after implantation of the breast prosthesis, augmentation mammaplasty through an inframammary incision and the external oblique muscle has been performed in 96 cases since 1990. In 67 of 72 cases (93%), the cosmetic results are satisfactory with a natural contour and softness of the breast during the follow-up of more than 6 months. The capsule contracture rate is 2.8 percent. The advantage of the approach is the muscular coverage of almost the total surface of the implant that contributes to the excellent result and a low capsule contracture rate. This procedure can overcome the disadvantages of the transaxillary augmentation procedure, which is usually difficult in dissecting the origin of the pectoralis muscle at the sixth rib. This technique can also make the inframammary crease lower and improve moderate ptosis of the breast.

Adult↗

[Effects of anabolics on the thermostimulated current spectrum in calf muscles].

The effects of growth substances (zeranol, trenbolone) on calf muscle (pectoralis transversus) have been studied with a recent physical method (thermostimulated current spectroscopy). This method appears promising for detecting meat from calves treated with such hormonal substances. The authors have hypothesized that the observed differences are related to protein modification, the nature of which is yet unknown. A complementary work is now in progress.

Animals↗