Attenuation of ultrasound in skeletal muscle

Ultrasonic attenuation in fresh and 5% formalin fixed beef skeletal muscle has been measured, as a continuous function of frequency, in the range 1–8 MHz, for muscle fibre orientations both parallel and normal to the direction of propagation. Good agreement was found in all cases between two independent sets of measurements employing transmission and reflection techniques respectively. The data are consistent with a power law dependence of attenuation coefficient on frequency, with an exponent that is not significantly different from unity. For propagation normal to the fibres attenuation values are found as 1.1 ± 0.15 and 1.6 ± 0.15 dB cm^?1 MHz^?1 for fresh and fixed tissue respectively, the corresponding values for parallel propagation being 2.9 ± 0.23 and 4.1 ± 0.25 dB cm^?1 MHz^?1.     

Quantitative assessment of skeletal muscle activation using muscle functional MRI

The purpose of the present study is to determine whether muscle functional MRI (mfMRI) can be used to obtain three-dimensional (3-D) images useful for evaluating muscle activity, and if so, to measure the distribution of muscle activity within a medial gastrocnemius (MG) muscle. Seven men performed 5 sets of 10 repetitions of a calf-raise exercise with additional 15% of body-weight load. Magnetic resonance images were obtained before and immediately after the exercise. To threshold images, only those pixels showing transverse relaxation time (T2) greater than the mean+1 S.D. of the entire regions of interest (ROIs) in the preexercise image and T2 lower than the mean+1 S.D. of the entire ROIs in the postexercise image were identified. The survived pixels showing T2 are defined as active muscle. Those thresholded images were 3-D reconstructed, and this was used to determine area of active muscle along transverse, longitudinal and vertical axes. At the exercise level used in the present study, the percentage volume of activated muscle in the MG was 62.8+/-4.5%. There was a significant correlation between percentage volume of activated muscle and integrated electromyography (r=.78, P<.05). Percentage areas of activated muscle were significantly larger in the medial than in the lateral region, in the anterior than in the posterior region and in the distal than in the proximal region (P<.05). These results suggest that mfMRI can be used to evaluate the muscle activity and to determine intramuscular variations of activity within skeletal muscle.     

Cryopreservation of human skeletal muscle impairs mitochondrial function

Previous studies have investigated if cryopreservation is a viable approach for functional mitochondrial analysis. Different tissues have been studied, and conflicting results have been published. The aim of the present study was to investigate if mitochondria in human skeletal muscle maintain functionality after long term cryopreservation (1 year). Skeletal muscle samples were preserved in dimethyl sulfoxide (DMSO) for later analysis. Human skeletal muscle fibres were thawed and permeabilised with saponin, and mitochondrial respiration was measured by high-resolution respirometry. The capacity of oxidative phosphorylation was significantly (P < 0.05) reduced in cryopreserved human skeletal muscle samples. Cryopreservation impaired respiration with substrates linked to Complex I more than for Complex II (P < 0.05). Addition of cytochrome c revealed an increase in respiration indicating cytochrome c loss from the mitochondria. The results from this study demonstrate that normal mitochondrial functionality is not maintained in cryopreserved human skeletal muscle samples.     

Attenuation of ultrasound in post rigor bovine skeletal muscle

A pulse transmission method for measuring the attenuation of 1-7 MHz ultrasound in bovine skeletal muscle is described. Measurements of the attenuation coefficient at -20, 0, 20 and 40 degrees C conformed to the relation alpha = Afn, where A and n are temperature-dependent coefficients and f is the frequency. alpha/f varied slowly with frequency, and at 4 MHz and 20 degrees C mean values were 1.3 dB cm-1 MHz-1 along the fibres and 0.55 dB cm-1 MHz-1 across the fibres. These data are lower than most previous measurements of skeletal muscle, but comparable with recent measurements of canine heart muscle.     

Polarization gating in ultrafast-optics imaging of skeletal muscle tissues

By comparing the results of polarization-dependent, time-resolved intensity profiles of photons transmitted through diluted milk, chicken breast tissue, and chopped chicken breast tissue, we found that the inherent anisotropic optical property of skeletal muscle tissue resulted in coherent coupling between two mutually perpendicular polarization directions. This coupling process led to difficulty in using the conventional polarization gating method for imaging unless the anisotropy characteristics were well understood. However, imaging based on polarization gating in diluted milk and chopped chicken breast tissue, which had an isotropic random-scattering nature, was quite effective.     

细胞生物力学

文章评述了组成细胞的各种材料与亚结构的力学特性,细胞及其分子所产生的力和细胞对力作用的响应,介绍了描述细胞形态结构与力学行为的本构方程和研究细胞力学的有关实验技术.内容不仅包括有关细胞生物力学各种主要及最新的研究成果,而且还包括作者对细胞力学的有关实验与理论研究结果,还有许多可供从事生命科学研究人员研究参考的有关细胞生物力学详实而又系统的资料和数据.     

细胞生物力学

文章评述了组成细胞的各种材料与亚结构的力学特性，细胞及其分子所产生的力和细胞对力作用的响应，介绍了描述细胞形态结构与力学行为的本构方程和研究细胞力学的有关实验技术．内容不仅包括有关细胞生物力学各种主要及最新的研究成果，而且还包括作者对细胞力学的有关实验与理论研究结果，还有许多可供从事生命科学研究人员研究参考的有关细胞生物力学详实而又系统的资料和数据．     

Oxygen-induced changes in longitudinal relaxation times in skeletal muscle

The objective was to measure the effect of 100% oxygen inhalation on T1 relaxation times in skeletal muscle. Healthy volunteers were scanned using three different MRI protocols while breathing medical air and 100% oxygen. Measurements of T1 were made from regions of interest (ROIs) within various skeletal muscle groups. Dynamic data of subjects breathing a sequence of air-oxygen-air allowed the calculation of characteristic wash-in and -out times for dissolved oxygen in muscle. Contrary to previous findings, a statistically significant decrease in T1 in skeletal muscle was observed due to oxygen inhalation. We report approximate baseline characteristic values for the response of skeletal muscle to oxygen inhalation. This measurement may provide new biomarkers for evaluation of oxygen delivery and consumption in normal and diseased skeletal muscle.     

Ultrasound speed and attenuation in homogenates of bovine skeletal muscle

The attenuation and speed of ultrasound were measured in homogenates of post-rigor bovine skeletal muscle, and found to increase in proportion to the concentration of muscle. Extrapolation of the data to tissue concentrations yielded an attenuation of 7.5 dB cm-1 at pH 5.7, 20 degrees C and 7.3 MHz. This was close to that measured in the minced tissue, 8.3 dB cm-1, and between values previously recorded across and along the fibres of intact muscle. Corresponding measurements for the speed of ultrasound in homogenates, extrapolated to the native tissue concentration, were: 1555 +/- 9 m s-1 at 0 degree C, 1592 +/- 10 m s-1 at 20 degrees C and 1616 +/- 9 m s-1 at 37 degrees C. These were not significantly different from measurements of minced muscle at the same temperatures. Measurements of the attenuation of 7.3 MHz ultrasound in suspensions of myofibrils indicated that attenuation by the myofibrils caused at least 64% of the attenuation in muscle homogenates at pH 5.7. Re-analysis of the viscous loss arising from relative movement of the myofibrils in their surrounding fluid, indicated that this mechanism could account for no more than 15% of the attenuation in muscle homogenates. Attenuation due to scattering was calculated to be at least two orders of magnitude smaller than that observed in either homogenates or suspensions of myofibrils. It was concluded that the contribution of scattering to the attenuation was small, and that the attenuation was caused by processes involving an absorption of energy.(ABSTRACT TRUNCATED AT 250 WORDS)     

Simultaneous measurements of diffusion and transverse relaxation in exercising skeletal muscle

The aim of this study was to compare proton T₂ and apparent diffusion coefficient (ADC) variations induced by exercise in skeletal muscle, to provide some more information on the source of their variations. T₂ and ADC were measured in the forearm flexor digitorum muscles in 12 healthy volunteers at rest and after an exercise, using a sequence allowing simultaneous measurements of both parameters. At rest, T₂ was 30.6 ± 1.8 ms (mean ± 1 SD) and ADC was 1.82 ± 0.11 × 10⁻⁹ m²/s. With exercise, T₂ varied by +28 ± 12% (p < .001 vs. rest) and ADC varied by +12 ± 3% (p < .001). The recovery of T₂ after exercise was faster than that of ADC, with half-times of 7 ± 2 min and of 15 ± 8 min (p < .01), respectively. It is concluded that both T₂ and ADC increase with exercise. However, the mechanisms of variation of T₂ and ADC with exercise are probably different, T₂ mostly reflecting changes in water content and ADC reflecting temperature variations.     

Changes in skeletal muscle diffusion parameters owing to intramyocellular lipid

IntroductionSeveral studies investigated the changes in diffusion of water molecules in skeletal muscle cells of lifestyle-related-disease patients who performed a hybrid training (HYBT) for six months. They reported that the apparent diffusion coefficient (ADC) and all diffusion eigenvalues (λ₁, λ₂, and λ₃) increased after the HYBT, owing to the enlargement of the intramyocellular diffusion space (intracellular space) caused by the muscular hypertrophy.We assumed that the HYBT promoted metabolism of the whole skeletal muscle including lipids, which reduced the amount of intramyocellular lipid (IMCL), and led to a secondary enlargement of the diffusion space in the skeletal muscle cells. However, the IMCL has to be a diffusion limiting factor in order to verify this hypothesis. Until now, there is no report on whether IMCL is a diffusion limiting factor for water molecules.The objective of this study was to examine whether the IMCL is a diffusion limiting factor in skeletal muscle cells.Materials and methodsWe performed a three-dimensional quantification of the IMCL in triceps surae muscles of lifestyle-related-disease patients and healthy volunteers. In addition, we measured the ADC in the volume of interest (VOI), diffusion anisotropy (FA), and diffusion eigenvalues (λ₁, λ₂, and λ₃), and evaluated the correlations between these diffusion parameters and IMCL.ResultsThe results showed that the amount of IMCL was positively and negatively correlated with the FA and λ₃, respectively, in lifestyle-related-disease patients. In addition, there was a weak negative correlation between IMCL and ADC, λ₁, and λ₂. There was no correlation between the amount of IMCL and diffusion parameters of healthy volunteers.DiscussionAbove a certain amount, the IMCL correlates with the diffusion parameters. A higher amount of IMCL leads to smaller diffusion eigenvalues. This result suggested that IMCL possibility of influencing diffusion of water molecules in skeletal muscle cells. However, in order for the influence of IMCL to be reflected in the diffusion eigenvalues, it was needed large amount of IMCL existed, and we thought that the influence was smaller than the influence by the already reported cell membrane.     

A novel BDNF gene promoter directs expression to skeletal muscle

Background  

Cell-specific expression of the gene that encodes brain-derived neurotrophic factor (BDNF) is required for the normal development of peripheral sensory neurons and efficient synaptic transmission in the mature central and peripheral nervous system. The control of BDNF gene expression involves multiple tissue and cell-specific promoters that are differentially regulated. The molecular mechanisms that are responsible for tissue and cell-specific expression of these promoters are still incompletely understood.     

Attenuation of ultrasound in homogenates of bovine skeletal muscle and other tissues

The attenuation of ultrasound in homogenates of bovine skeletal muscle and suspensions of myofibrils was measured over the frequency range 1.5-7 MHz, and found to be proportional to protein concentration in both. In the homogenates it varied with frequency and temperature in a similar way to the attenuation in post rigor muscle tissue; myofibrils showed a higher frequency dependence. The attenuation in homogenates of bovine muscle, liver and kidney and in suspensions of myofibrils was measured over the pH range 3.5-13, and each showed a peak at about pH 11.5. This was thought to be due to a proton transfer process between NH3+ groups on the tissue proteins and OH- ions in the suspending fluid. A substantial peak at about pH 5 in the muscle and myofibril suspensions was not observed in homogenates of liver and kidney and was thought to be due to components of muscle that are absent from the other tissues. Myofibrils suspended in percoll solution of density 1.05 g cm-3, chosen to match approximately the density of the myofibrils, showed a slightly lower attenuation over the pH range 5-7, but a pH dependence similar to that of the myofibrils suspended in saline. The difference in the attenuations may be interpreted as the viscous component of the attenuation due to relative motion between the myofibril and its surrounding saline. The peak at pH 5 did not, however, appear to be due to the viscous loss mechanism peaking due to maximum shrinkage (and therefore maximum density) of the myofibril near this pH.(ABSTRACT TRUNCATED AT 250 WORDS)     

Experimental estimation of the viscous component of ultrasound attenuation in suspensions of bovine skeletal muscle myofibrils

It has been suggested that viscous losses, caused by the motion of myofibrils relative to their environmental fluid, could be the major cause of ultrasound attenuation in muscle. This Paper presents theoretical and experimental estimates of the viscous component of attenuation in suspensions of myofibrils. Experimental estimates were made by monitoring the effects of varying the viscosity and density of the suspending fluid and ranged from 0.55 to 0.72 cm2 g-1 protein at 7 MHz, when expressed as mass attenuation coefficients. This represented 16-22% of the total attenuation measured in suspensions. Corresponding theoretical calculations were lower: 0.44 and 0.21 cm2 g-1 protein, respectively, for suspensions with the myofibrils aligned along and across the direction of sound propagation. It was concluded that most of the attenuation was caused by other absorption processes.     

Effect of exercise on the creatine resonances in 1H MR spectra of human skeletal muscle

1H MR spectra of human muscles were recorded before, during, and after fatiguing exercise. In contrast to expectations, it was found that the spectral contributions of creatine/phosphocreatine (Cr/PCr) were subject to change as a function of exercise. In particular, the dipolar-coupled methylene protons of Cr/PCr were found to be reduced in intensity in proportion to the co-registered PCr levels. Recovery after exercise and behavior under ischemic conditions provide further evidence to suggest that the contributions of the CH2 protons of Cr/PCr to 1H MR spectra of human muscle in vivo reflect PCr rather than Cr levels. Variation of experimental parameters showed that this effect is not due to a trivial change in relaxation times. At present it can only be speculated about why the Cr resonances have reduced NMR visibility. If temporary binding to macromolecules should be involved, the free Cr concentration-important for equilibrium calculations of the creatine kinase reaction-might be different from what was previously assumed.     

Early ultrastructural events of skeletal muscle damage following cardiotoxin-induced injury and glycerol-induced injury

In this study, we investigated the early changes of skeletal muscle damage in response to injuries induced by cardiotoxin (CTX) and glycerol by using both light microscopy and transmission electron microscopy. Normal, non-dystrophic, adult male mice were used in this study. Tibialis anterior (TA) muscles were injected either with CTX or glycerol. Samples were collected at intervals starting from 1 h up to 4 days after injury. Injured muscles were subjected to both histological and ultrastructural analyses. CTX-induced injury caused mitochondrial accumulation and swelling followed by lysis, while glycerol-induced injury caused accumulation of vesicles with focal disruption of the basal lamina, indicating that the injuries have different mechanisms of damage to myofibers. Moreover, inflammatory cells, including neutrophils and macrophages, were recruited earlier and in larger numbers after CTX-induced injury than after glycerol-induced injury. On the other hand, satellite cells (SCs) activation started at 6 h after both injuries, as indicated by an increase in both the length and cytoplasmic-to-nuclear ratio. However, there were significantly longer SCs with a higher cytoplasmic-to-nuclear ratio in the CTX-injured muscles than in the glycerol-injured muscles at day 4. In conclusion, our results demonstrated a difference between CTX and glycerol in their damage to myofibers; CTX damages myofiber mitochondria, while glycerol damages the myofiber cell membrane and alters osmosis. In addition, CTX-induced injury caused earlier and more extensive inflammatory infiltration than did glycerol-induced injury. This study is the first study to shed light on the early events following skeletal muscle injury induced by CTX and glycerol.     

NMR relaxation times of skeletal muscle: Dependence on fiber type and diet

Skeletal muscles are composed of a mixture of different types of muscle fibers. Previous NMR studies of muscle have typically used muscles of mixed fiber composition and have not taken into account factors such as the dietary history of the experimental animal. In this paper we present evidence that the T₁ and T₂ relaxation times of two of the major types of skeletal muscle fiber are not significantly different under normal conditions. Following dietary manipulation of the rabbits (limiting potassium intake or increasing cholesterol consumption), the T₁ and T₂ relaxation times of psoas and soleus muscles were significantly different. The change in relaxation times of psoas muscles following dietary manipulation can be only partially explained by shifts in muscle water content. The results suggest that changes in diet are capable of inducing changes in water bonding and structuring in muscle tissues. Our results indicate that diet must be added to the growing list of environmental factors that can cause NMR contrast changes. Also, NMR measurements of muscles rich in one fiber type or another could be of value for detecting changes in body composition.     

Surface and inner cell behaviour along skeletal muscle cell in vitro differentiation

During muscle tissue differentiation, in particular in the formation of myotubes from the myoblasts, plasma membrane changes its morpho-functional characteristics. In this study, muscle cell membrane behaviour has been studied along the differentiation of C2C12, a mouse myoblastic adherent cell line. Flat undifferentiated cells, cultured for 3-4 days in the differentiation medium, progressively become thick, long and multinucleated myotubes covered with microvilli. They lose stress fibers and adhesion to the underlying substrate evidentiating an actin redistribution, followed by the spatial organization of thick and thin myofilaments. Sarcomeres and myofibrils occasionally appear, even if a certain percentage of "myosacs" containing randomly oriented filaments can be identified all along the differentiation. M-cadherin, a molecule involved in cell-cell adhesion, also appears in the early differentiation stage, during myoblast fusion. Occasional focal contractions can also be observed in myotubes, which prompt an electrophysiological membrane analysis. When studied by means of patch clamp technique, resting membrane potential appears to undergo a transient depolarization, while input resistance increases until day 5 after differentiation induction, then successively decreases. Capacitance declines until day 5, later appearing enhanced. Moreover, with the induction of differentiation, the pattern of functional voltage-dependent ion channels changes. Therefore, during myogenesis, cell maturation is coupled with changes in cell membrane morphological features and functional characteristics.     

Localized in vivo H spectroscopy of human skeletal muscle: Normal and pathologic findings

To obtain high signal to noise ratio in small volume elements (8 cm³), in vivo ¹H NMR spectroscopy of normal and diseased human skeletal muscle was performed using a double spin-echo localization method on a 1.5-T whole body system. High resolved spectra of normal calf muscle show the well known resonances of lipids (methyl, methylene, olefinic, and other fatty acid resonances), creatine/phosphocreatine, choline/carnitine, taurine, and histidine with good intraindividual reproducibility. Pronounced intraindividual differences in the lipid range were found between different upper thigh muscle groups. On pathologic conditions like myopathia, myositis or irradiation damage the spectral lipid content was increased. Three months after local irradiation of the medial vastus muscle (50 Gy), the localized ¹H NMR spectrum showed a complete loss of the choline and creatine signals. In a case of M. Behçet with muscular involvement the relative reduction of the choline signal may provide an insight in the pathobiochemistry. The results of our investigations in nine healthy volunteers and three patients are presented in detail including relaxation times of the metabolites.     

Anisotropy of ultrasonic propagation and scattering properties in fresh rat skeletal muscle in vitro

The anisotropy of frequency-dependent backscatter coefficient, attenuation, and speed of sound is assessed in fresh rat skeletal muscle within 5 h post-mortem. Excised rat semimembranosus and soleus muscles are measured in 37 degrees C Tyrode solution, with the muscle fibers at 90 degrees and 45 degrees orientations to the incident sound beam. Reflected and through transmission signals from either a 6- or 10-MHz focused transducer give frequency dependent information in the 4-14 MHz range. The attenuation coefficient in each muscle is consistently a factor of 2.0 +/- 0.4 lower for propagation perpendicular to the fibers than at 45 degrees, whereas speed of sound shows a much milder anisotropy, and is slightly faster for the 90 degrees orientation. The largest anisotropy is seen in the backscatter coefficient, most notably in the semimembranosus where the magnitude at 90 degrees is over an order of magnitude greater than at 45 degrees, with the frequency dependence in both cases giving a power law between 1.5 and 2.0.