Assessment of muscle volume and physiological cross-sectional area of the human triceps surae muscle in vivo

The purpose of the present study was to investigate whether it is possible to predict the individual muscle volumes within the triceps surae (TS) muscle group by means of easily measurable parameters based on a theoretical consideration. A further objective was to verify the use of the available literature data to assess the contribution of each muscle of the group to the entire TS volume or physiological cross-sectional-area (PCSA). Therefore, magnetic resonance images of the right calf of 13 male subjects were acquired and each muscle of the TS was reconstructed. Muscle length (l(m)), the maximum anatomical cross-sectional-area (ACSA(max)) and muscle volume were obtained from the 3D models. To assess the PCSA, fascicle length was determined by ultrasonography. In general, muscle volume can be expressed as a fraction of the product of ACSA(max) and l(m). The size of the fraction depends on muscle shape and its coefficient of variance among the examined population was considerable low (soleus 6%, gastrocnemius 4% and gastrocnemius lateralis 7%) in the present study. The product of ACSA(max) and l(m) was, therefore, suitable to assess muscle volume (root mean squares, RMS 4-7%). Further, the soleus, gastrocnemius medialis and gastrocnemius lateralis accounted on average for about 52+/-3%, 32+/-2% and 16+/-2% of the total TS volume and 62+/-5%, 26+/-3% and 12+/-2% of the entire TS PCSA, respectively. The coefficient of variance of the relative portions were 5-10% for muscle volume and 8-17% for the PCSA.     

MicroRNA Transcriptome Profile Analysis in Porcine Muscle and the Effect of miR-143 on the MYH7 Gene and Protein

Porcine skeletal muscle fibres are classified based on their different physiological and biochemical properties. Muscle fibre phenotype is regulated by several independent signalling pathways, including the mitogen-activated protein kinase (MAPK), nuclear factor of activated T cells (NFAT), myocyte enhancer factor 2 (MEF2) and peroxisome proliferator-activated receptor (PPAR) signalling pathways. MicroRNAs are non-coding small RNAs that regulate many biological processes. However, their function in muscle fibre type regulation remains unclear. The aim of our study was to identify miRNAs that regulate muscle fibre type during porcine growth to help understand the miRNA regulation mechanism of fibre differentiation. We performed Solexa/Illumina deep sequencing for the microRNAome during 3 muscle growth stages (63, 98 and 161 d). In this study, 271 mature miRNAs and 243 pre-miRNAs were identified. We detected 472 novel miRNAs in the muscle samples. Among the mature miRNAs, there are 23 highest expression miRNAs (over 10000 RPM), account for 85.3% of the total counts of mature miRNAs., including 10 (43.5%) muscle-related miRNAs (ssc-miR-133a-3p, ssc-miR-486, ssc-miR-1, ssc-miR-143-3p, ssc-miR-30a-5p, ssc-miR-181a, ssc-miR-148a-3p, ssc-miR-92a, ssc-miR-21, ssc-miR-126-5p). Particularly, both ssc-miR-1 and ssc-miR-133 belong to the MyomiRs, which control muscle myosin content, myofibre identity and muscle performance. The involvement of these miRNAs in muscle fibre phenotype provides new insight into the mechanism of muscle fibre regulation underlying muscle development. Furthermore, we performed cell transfection experiment. Overexpression/inhibition of ssc-miR-143-3p in porcine skeletal muscle satellite cell induced an/a increase/reduction of the slow muscle fibre gene and protein (MYH7), indicating that miR-143 activity regulated muscle fibre differentiate in skeletal muscle. And it regulate MYH7 through the HDAC4-MEF2 pathway.     

Effect of temperature on the interaction of gramicidin A and its dimer analog with the muscle fiber membrane

Potassium conductance of single muscle fibres from Rana esculenta was studied in isotonic K2SO4 solution under constant current conditions using double sucrose gap method. At room temperature the channel formation by gramicidin was much faster than that of the synthetic head to head covalently linked gramicidin dimer. The increase of temperature by 8-10 degrees C resulted in a considerable rise of both dimer- and gramicidin-induced conductances. The effect was much greater than in the case of bilayers indicating a remarkable entropy change in the muscle fibre membrane. The temperature dependence of adsorption was more pronounced than that of desorption: there was no effect on desorption of dimer and only 20% of the temperature-activated desorption of gramicidin irreversibly bound at room temperature.     

Estimation of errors in mechanical efficiency

Errors in measurements of mechanical work, net energy expenditure and mechanical efficiency (ME) were calculated, when subjects performed isolated eccentric or concentric muscle actions and combinations of these actions [stretch-shortening cycle (SSC) exercises] with a special sledge apparatus. The relative error of mechanical work was 6.1%. When estimating the error of energy metabolism from oxygen consumption the error would be about 4% (McArdle et al. 1981). The maximum error of ME was the sum of these two values (10.1%). Obviously the error of ME was less than 5%, because 30 muscle actions were averaged and, in addition, the errors of mechanical work and energy expenditure were not in the same direction every time. It was concluded that mechanical work can be determined accurately when the force is measured as a function of the moved distance of the sledge. Thus calculation of ME can be performed quite reliably in isolated eccentric and concentric exercises. The greatest problems were, however, in the SSC exercises, where the errors were higher, because of the problems of dividing the net energy expenditure into eccentric and concentric phases. Therefore, further developments must be made to minimize the errors in measurement and calculation during SSC-exercise.     

Effect of temperature on the inward rectifier and gramicidin A-induced channels in the membrane of frog skeletal muscle fibres

The conductance of frog skeletal muscle fibres in isotonic K2SO4 solution has been measured. Experiments were carried out under current-clamp conditions using a double sucrose-gap technique. The potassium conductances of the inward rectifier and the gramicidin channel in the same muscle fibre were compared. Potassium conductance of the inward rectifier increased with the temperature, with a value of Q10 1.55 +/- 0.09 (n = 8) under hyperpolarization, and Q10 2.38 +/- 0.23 (n = 6) for the depolarizing stimulus, the difference between Q10 of potassium and gramicidin channels being statistically insignificant.     

The effect of vanadate on the electrogenic Na+/K+ pump, intracellular Na+ concentration and electrophysiological characteristics of mouse skeletal muscle fibre

The present study reports a discrepancy between the effects of vanadate on the membrane Na+-K+-ATPase and the Na+/K+ pump of the skeletal muscle. Vanadate in concentration 4 X 10(-6) mol/l which is necessary to block the enzyme Na+-K+-ATPase activity of membrane fractions failed to inhibit the electrogenic Na+/K+ pump of intact muscle cells. The effect of vanadate on the electrophysiological parameters of the muscle fibre membrane required much higher vanadate levels, but again, Na+/K+ pump was still active. Vanadate in concentrations 4 X 10(-4) and 4 X 10(-5) mol/l depolarized the membrane potential and decreased the membrane resistance [apparently in consequence of enhanced passive membrane permeability for Na+ ions]. Action potentials and the electrical excitability of the muscle fibre membrane were reduced by these vanadate concentrations.     

Relationship between the physiological cross-section of the human jaw muscles and their cross-sectional area in computer tomograms

Physiological cross-section (PCS) and cross-sectional area in computer tomograms made at right angles to the mean fibre direction were compared in the masseter, temporalis and pterygoid muscles of six human cadavers. PCS was determined as (1) total cross-section of teased fibre bundles (2) total fibre weight divided by mean fibre length. The two measures correlated strongly, but the first was always 25% lower than the second, irrespective of the muscle concerned. The cross-sections in the tomograms (SCS) were smaller than the PCS, except in the lateral pterygoid. In all muscles, a statistically significant correlation was found between SCS and PCS. The SCS can be used to predict PCS, with an error of 0.3-1.0 cm2. In our material, cross-sections were about 20% higher than reported in the literature. It is suggested, that this discrepancy is caused by the loss of natural teeth.     

Morphological changes in continuously stretched skeletal muscles in sheep

The effects of continuous elongation of skeletal muscles were studied on six sheep who underwent a lengthening osteotomy of the right tibia. Open muscle biopsies were taken from the biceps femoris muscle preoperatively (Group A), after 5 weeks of bone distraction (Group B) and after another 5 weeks without further distraction (Group C). The size and distribution of type 1 (slow-twitch) and type 2 (fast-twitch) muscle fibres were determined from sections stained for myofibrillar ATPase activity. All sections were also evaluated by light microscopy, especially with regard to myopathic changes. The type 2 fibres showed a significant decrease in size from group A to B and from group B to C. The reduction in fibre size from group A to C was 44.2%. The type 1 fibres, on the other hand, showed no significant differences in mean fibre size between the groups. However, there were considerable individual variations in type 1 fibre size between the groups. The distribution of both fibre types was similar in groups A and B (appr. 17% type 1 fibres) whereas the relative number of type 1 fibres was reduced to 12.4% in group C (P less than 0.01). Myopathic changes, i.e. muscle fibre necroses, were not seen in any of the groups. It is concluded that the type 2 fibre atrophy is mainly caused by muscular inactivity during the postoperative period, but an additional effect of continuous stretching of the muscle cannot be excluded.(ABSTRACT TRUNCATED AT 250 WORDS)     

Muscle architecture and fibre characteristics of rat gastrocnemius and semimembranosus muscles during isometric contractions

Rat gastrocnemius medialis (GM) and semimembranosus (SM) muscles have a very different morphology. GM is a very pennate muscle, combining relatively short muscle fibre length with sizable fibre angles and long muscle and aponeurosis lengths. SM is a more parallel-fibred muscle, combining a relatively long fibre length with a small fibre angle and short aponeurosis length. The mechanisms of fibre shortening as well as angle increase are operational in GM as well as SM. However, as a consequence of isometric contraction, changes of fibre length and angle are greater for GM than for SM at any relative muscle length. These differences are particularly notable at short muscle lengths: at 80% of optimum muscle length, fibre length changes of approximately 30% are coupled to fibre angle changes of 15 degrees in GM, while for SM these changes are 4% and 0.6 degrees, respectively. A considerable difference was found for normalized active slack muscle length (GM approximately 80 and SM approximately 45%). This is explained by differences of degree of pennation as well as factors related to differences found for estimated fibre length-force characteristics. Estimated normalized active fibre slack length was considerably smaller for SM than for GM (approximately 40 and 60%, respectively). The most likely explanation of these findings are differences of distribution of optimum fibre lengths, possibly in combination with differences of myofilament lengths and/or fibre length distributions.     

Heterogeneous response of isolated adult rat heart cells to insulin

3-O-Methylglucose uptake by Ca2+-resistant adult rat heart cells in suspension was measured, free of artifactual inhibitor-insensitive uptake, and with an accuracy of +/- 1.9% pellet water. (Ca2+-resistant cells are cells which retain their original rod-shaped morphology in the presence of physiological levels of Ca2+.) High levels of insulin (10(-6) M) stimulated the rate of 3-O-methylglucose uptake approximately 10-fold. In the presence of low levels of insulin (3 X 10(-11) M, 10(-10) M) uptake was biphasic; it could not be described by a single exponential function within experimental error, but required the sum of two exponentials. Deviation from a single exponential function was not so great with high levels of insulin (10(-6) M) or no insulin. Cell sugar uptake was also investigated using autoradiography of cells which had accumulated [2-14C]deoxyglucose under similar conditions. This showed considerable heterogeneity of 2-deoxyglucose uptake by cells treated with low levels of insulin, but significantly less heterogeneity of 2-deoxyglucose uptake by cells treated with high levels of insulin. It is concluded that the deviation of 3-O-methylglucose uptake from a single exponential observed at low insulin levels can be accounted for in terms of a heterogeneous response of cells to insulin.     

Relative contribution of gravity to pulmonary perfusion heterogeneity.

We designed a series of experiments and analyses to quantify the contribution of gravity to pulmonary perfusion heterogeneity. Regional pulmonary perfusion was measured in five anesthetized and ventilated dogs in both supine and prone positions by use of radiolabeled microspheres injected during apnea at functional residual capacity. Measurements of flow were repeated in each position, and the sequence of positions was prospectively designed to nullify any effect of order. The lungs of each animal were excised, perfused with saline until clear, dried at an inflation pressure of 25 cmH2O, and cut into 1.9-cm3 pieces. Each piece was weighed and the radioactivity determined in a scintillation counter. Measurement errors were minimized by excluding lung pieces that had greater than 25% airway and weighed less than 10 mg or greater than 60 mg. Weight-normalized flows in each position and repetition were determined for each lung piece. An analysis of variance model was used to identify the percentage of variation in regional flow that was due to position (supine vs. prone), to random error and time (measurement and repetition), and to structure, where structure was defined as the component of flow that remained constant across position and replication. The contributions of position, error/time, and structure to the total variability of flow across the five dogs were 7.8 +/- 0.6, 8.4 +/- 8.3, and 83.8 +/- 8.4%, (SD), respectively. Because the contribution of position represents the additive effect of gravity between two opposite positions, the contribution of gravity to perfusion heterogeneity in one position may be as little as 4%.(ABSTRACT TRUNCATED AT 250 WORDS)     

Blood flow distribution and tissue allometry in channel catfish

Blood flow (as percentage of cardiac output) in fasted channel catfish acclimated to 21°C was directed primarily to white muscle (72%) followed by head kidney (5·7%), red muscle (5·5%), trunk kidney (3·1%), liver (2·2%), swim bladder (1·4%) and skin (1·1%). The stomach, intestines, pyloric caeca, gonads, brain, abdominal fat and spleen contained <0·5% of blood flow. There was considerable interfish variation among blood flow distribution to visceral organs with substantial spatial heterogeneity of blood flow to white muscle. The spatial heterogeneity of flow to muscle prevented accurate estimation of total flow to this tissue based on the microsphere deposition of a few sub-samples. Instead, a novel approach, based on the whole animal counting of the eviscerated carcass was used to measure blood flow to white muscle. The scaling relationships for tissue mass in catfish (63–1873 g) followed the allometric equation (aW^b) and tended to exhibit negative allometry, with organ weight decreasing in proportion to body weight. The b values for most tissues ranged between 0·83 and 1·0. The relative mass of the brain showed the greatest decline and with a b value of 0·32. The results, together with previous data on cardiac output, permitted calculation of organ blood flow rates in channel catfish. © 1999 The Fisheries Society of the British Isles     

Cementum annulation and age determination in Homo sapiens. I. Tooth variability and observer error

In order to test the feasibility of cementum annulations to estimate age in humans, observer error and tooth variability in cementum ring counts were evaluated in a sample of 42 mandibular canine and first premolar pairs. Additionally, two sectioning techniques were evaluated. Demineralized thin sections (7 micron) stained with hematoxylin are the preferred technique since their age related variance is greater than 75% for all tooth types examined. In contrast, less than 50% of the total variance was accounted for among individuals when mineralized sections (80 micron) stained with alizarin red were used. Intertooth variability in ring counts of demineralized sections was large between canines and premolars (43%). Premolars provide counts with lower interobserver error and are the preferred tooth. In an expanded sample (N = 51) of demineralized premolars, intraobserver and interobserver error accounted for 2% and 5% of the total variance, respectively. Evaluation of several experimental designs showed that increasing the number of slides per tooth has the greatest effect on reducing variance followed by increasing the number of observers. Increasing the number of observations has little effect. Cementum ring counts are measurable to a highly repeatable extent and provide a level of repeatability greater than that reported for the pubic symphysis and auricular surface aging techniques.     

Influence of muscle geometry on shortening speed of fibre, aponeurosis and muscle.

The influence of muscle geometry on muscle shortening of the gastrocnemius medialis muscle (GM) of the rat was studied. Using cinematography, GM geometry was studied during isokinetic concentric activity at muscle lengths ranging from 85 to 105% of the optimum muscle length. The shortening speed of the distal fibre, the proximal aponeurosis and the muscle were determined, as well as the effect of rotation of the distal fibre and the proximal aponeurosis on the muscle speed of shortening. The results show that, due to the geometrical configuration, muscle shortening speed is not only determined by the speed of the fibre, but also to a large extent by the aponeurosis shortening speed. At optimum muscle length, the fibre and aponeurosis shortening speeds expressed relative to the muscle shortening speed amounted to 84% and 6%, respectively. At shorter muscle length, fibre speed relative to muscle speed decreased to values as low as 35%, whereas that of aponeurosis increased to values as high as 31%. Angular effects on the muscle speed of shortening can explain 10% of the muscle shortening speed at optimum muscle length and up to 34% of the muscle speed at shorter muscle length. In addition, a model was formulated to simulate the geometrical effects on muscle speed. This model, incorporating both fibre and aponeurosis length changes, contains a transfer function relating the shortening speeds of fibre and aponeurosis to muscle speed. The muscle shortening speed calculated using this transfer function demonstrated no significant differences with the speed measured experimentally.     

Correlation of electrophysiological,histochemical, and mechanical properties in fibres of the coxa rotator muscle of the locust,Locusta migratoria

Summary The fibre composition of the anterior coxa rotator muscle of the locust middle leg (M92) was examined. The muscle is composed of 90–100 fibres. Muscle fibres were characterized with regard to innervation pattern, electrophysiological properties, and morphological parameters. Activity and isoenzyme composition of myofibrillar ATPase, succinic acid dehydrogenase (SDH) activity and glycogen content were examined employing histochemical techniques. Shortening velocity and the dependence of tension on intracellular Ca²⁺ were determined in skinned fibre experiments. A close match was observed between the innervation pattern of the muscle fibres and their histochemical and physiological properties. The combination of all parameters examined allowed an accurate classification of the muscle fibres into three types. Within a given type, broad variability of some properties was observed (SDH activity, Ca²⁺ sensitivity) while others assumed distinct values (innervation pattern, shortening velocity). The comprehensive characterization of muscle fibre properties permits a functional interpretation of fibre heterogeneity with regard to muscle performance. Fibres with the same innervation pattern may be recruited specifically, according to their electric properties and Ca²⁺ sensitivities. The resulting specific recruitment of fibres with different mechanical responses should allow a subtle control of muscular force, with regard to force amplitude, temporal characteristics of contraction, and metabolic cost.Abbreviations CI₁ common inhibitory neurone one - ejp ijp excitatory, inhibitory junctional potential - EGTA ethylene glycol-bis[-aminoethyl ether] N,N,N,N-tetraacetic acid - mATPase myofibrillar adenosinetriphosphatase - MOPSO 3-[N-morpholino]-2-hydroxypropanesulfonic acid - M92 anterior rotator muscle of the coxa - n Hill coefficient - pCa₅₀ pCa corresponding to half-maximal tension - P₀ maximal isometric tension - SDH succinic acid dehydrogenase - V _max maximal shortening velocity     

Tension responses to increased hydrostatic pressure in glycerinated rabbit psoas muscle fibres

A method developed to study the effect of increased hydrostatic pressure on the isometric tension of a single muscle fibre is described and experiments done at room temperature (18-22 degrees C) on glycerinated rabbit psoas muscle fibres are presented. Increase of pressure (range 1-10 MPa) caused little change in tension transducer response when a muscle fibre was relaxed. However, there was a reversible depression of isometric tension with an increase of pressure when a fibre was maximally calcium-activated or in rigor; the depression was around 15% for active tension and 30% for rigor tension, for an increase of pressure of 10 MPa (ca. 100 atm).     

The turnover of cytochrome c in different skeletal-muscle fibre types of the rat.

The turnover of cytochrome c was determined in the three skeletal-muscle fibre types of adult male rats by a kinetic analysis that followed the time course of cytochrome c content change. Confirming evidence was obtained with double-labelling studies using delta-aminolaevulinate. Cytochrome c turnover was most rapid in the low-oxidative fast-twitch white fibre [t1/2 (half-life) about 4 days], slowest in the high-oxidative fast-twitch red fibre (t1/2 9-10 days) and relatively rapid in the high-oxidative slow-twitch red fibre (t1/2 5-6 days). Thus cytochrome c turnover does not strictly conform to either the appearance (i.e. red or white) or the contractile characteristics (i.e. fast or slow) of the muscle fibres. The synthesis rates needed to maintain the corresponding cytochrome c concentrations, however, were similarly high in the two mitochondria-rich red fibre types. These data illustrate that both the synthesis and degradation processes are important in establishing the cytochrome c concentrations that distinguish the different skeletal-muscle fibre types.     

Effect of innervation zones in estimating biceps brachii force-EMG relationship during isometric contraction

Measuring muscle forces in vivo is invasive and consequently indirect methods e.g., electromyography (EMG) are used in estimating muscular force production. The aim of the present paper was to examine what kind of effect the disruption of the physiological signal caused by the innervation zone has in predicting the force/torque output from surface EMG. Twelve men (age 26 (SD ±3) years; height 179 (±6) cm; body mass 73 (±6) kg) volunteered as subjects. They were asked to perform maximal voluntary isometric contraction (MVC) in elbow flexion, and submaximal contractions at 10%, 20%, 30%, 40%, 50% and 75% of the recorded MVC. EMG was measured from biceps brachii muscle with an electrode grid of 5 columns × 13 rows. Force-EMG relationships were determined from individual channels and as the global mean value. The relationship was deemed inconsistent if EMG value did not increase in successive force levels. Root mean squared errors were calculated for 3rd order polynomial fits. All subjects had at least one (4-52) inconsistent channel. Two subjects had inconsistent relationship calculated from the global mean. The mean root mean squared error calculated using leave one out method for the fits of the individual channels (0.33 ± 0.17) was higher (P < 0.001) than the error for the global mean fit (0.16 ± 0.08). It seems that the disruption of the physiological signal caused by the innervation zone affects the consistency of the force-EMG relationship on single bipolar channel level. Multichannel EMG recordings used for predicting force overcame this disruption.