Muscle fiber-type distribution, fiber-type-specific damage, and the Pompe disease phenotype.

Pompe disease is a lysosomal storage disorder caused by acid α-glucosidase deficiency and characterized by progressive muscle weakness. Enzyme replacement therapy (ERT) has ameliorated patients' perspectives, but reversal of skeletal muscle pathology remains a challenge. We studied pretreatment biopsies of 22 patients with different phenotypes to investigate to what extent fiber-type distribution and fiber-type-specific damage contribute to clinical diversity. Pompe patients have the same fiber-type distribution as healthy persons, but among nonclassic patients with the same GAA mutation (c.-32-13T>G), those with early onset of symptoms tend to have more type 2 muscle fibers than those with late-onset disease. Further, it seemed that the older, more severely affected classic infantile patients and the wheelchair-bound and ventilated nonclassic patients had a greater proportion of type 2x muscle fibers. However, as in other diseases, this may be caused by physical inactivity of those patients.

Deletion of the hypoxia-response element in the vascular endothelial growth factor promoter causes motor neuron degeneration.

Hypoxia stimulates angiogenesis through the binding of hypoxia-inducible factors to the hypoxia-response element in the vascular endothelial growth factor (Vegf) promotor. Here, we report that deletion of the hypoxia-response element in the Vegf promotor reduced hypoxic Vegf expression in the spinal cord and caused adult-onset progressive motor neuron degeneration, reminiscent of amyotrophic lateral sclerosis. The neurodegeneration seemed to be due to reduced neural vascular perfusion. In addition, Vegf165 promoted survival of motor neurons during hypoxia through binding to Vegf receptor 2 and neuropilin 1. Acute ischemia is known to cause nonselective neuronal death. Our results indicate that chronic vascular insufficiency and, possibly, insufficient Vegf-dependent neuroprotection lead to the select degeneration of motor neurons.

Ubiquitin-proteasome-dependent proteolytic activity remains elevated after zymosan-induced sepsis in rats while muscle mass recovers.

We studied the role of the ubiquitin-proteasome system in rat skeletal muscle during sepsis and subsequent recovery. Sepsis was induced with intraperitoneal zymosan injections. This model allows one to study a sustained and reversible catabolic phase and mimics the events that prevail in septic and subsequently recovering patients. In addition, the role of the ubiquitin-proteasome system during muscle recovery is poorly documented. There was a trend for increased ubiquitin-conjugate formation in the muscle wasting phase, which was abolished during the recovery phase. The trypsin- and chymotrypsin-like peptidase activities of the 20S proteasome peaked at day 6 following zymosan injection (i.e. when both muscle mass and muscle fiber cross-sectional area were reduced the most), but remained elevated when muscle mass and muscle fiber cross-sectional area were recovering (11 days). This clearly suggests a role for the ubiquitin-proteasome pathway in the muscle remodeling and/or recovery process. Protein levels of 19S complex and 20S proteasome subunits did not increase throughout the study, pointing to alternative mechanisms regulating proteasome activities. Overall these data support a role for ubiquitin-proteasome dependent proteolysis in the zymosan septic model, in both the catabolic and muscle recovery phases.

Skeletal muscle transverse strain during isometric contraction at different lengths.

An important assumption in 2D numerical models of skeletal muscle contraction involves deformation in the third dimension of the included muscle section. The present paper studies the often used plane strain description. Therefore, 3D muscle surface deformation is measured from marker displacements during isometric contractions at various muscle lengths. Longitudinal strains at superficial muscle fibers ( - 14 +/- 2.6% at L0, n = 57) and aponeurosis (0.8 +/- 0.9% at L0) decrease with increasing muscle length. The same holds for transverse muscle surface strains in superficial muscle fibers and aponeurosis, which are comparable at intermediate muscle length, but differ at long and short muscle length. Because transverse strains during isometric contraction change with initial muscle length, it is concluded that the effect of muscle length on muscle deformation cannot be studied in plane strain models. These results do not counteract the use of these models to study deformation in contractions with approximately - 9 % longitudinal muscle fiber strain, as transverse strain in superficial muscle fibers and in aponeurosis tissue is minimal in that case. Aponeurosis surface area change decreases with increasing initial muscle length, but muscle fiber surface area change is - 11%, independent of muscle length. Assuming incompressible muscle material, this means that strain perpendicular to the muscle surface equals 11%. Taking the relationship between transverse and longitudinal muscle fiber strain into account, it is hypothesized that superficial muscle fibers flatten during isometric contractions.

Spatial interaction between tissue pressure and skeletal muscle perfusion during contraction.

The vascular waterfall theory attributes decreased muscle perfusion during contraction to increased intramuscular pressure (P(IM)) and concomitant increase in venous resistance. Although P(IM) is distributed during contractions, this theory does not account for heterogeneity. This study hypothesises that pressure heterogeneity could affect the interaction between P(IM) rise and perfusion. Regional tissue perfusion during submaximum (100kPa) tetanic contraction is studied, using a finite element model of perfused contracting skeletal muscle. Capillary flow in muscles with one proximal artery and vein (SIM(1)) and with an additional distal artery and vein (SIM(2)) is compared. Blood flow and pressures at rest and P(IM) during contraction ( approximately 25kPa maximally) are similar between simulations, but capillary flow and venous pressure differ. In SIM(2), venous pressure and capillary flow correspond to P(IM) distribution, whereas capillary flow in SIM(1) is less than 10% of flow in SIM(2), in the muscle half without draining vein. This difference is caused by a high central P(IM), followed by central venous pressure rise, in agreement with the waterfall theory. The high central pressure (SIM(1)), obstructs outflow from the distal veins. Distal venous pressure rises until central blood pressure is reached, although local P(IM) is low. Adding a distal vein (SIM(2)) restores the perfusion. It is concluded that regional effects contribute to the interaction between P(IM) and perfusion during contraction. Unlike stated by the vascular waterfall theory, venous pressure may locally exceed P(IM). Although this can be explained by the principles of this theory, the theory does not include this phenomenon as such.

Spatial and temporal heterogeneity of superficial muscle strain during in situ fixed-end contractions.

Numerical models of contracting muscle offer a powerful tool to study local mechanical load. For validation of these models, the spatial and temporal distribution of strain was quantified in fixed-end contracting rat tibialis anterior muscle in situ at optimal muscle length (L(o)) and at 120 degrees plantar flexion as well as at 125 and 33Hz stimulation frequency. We studied the hypothesis that after termination of stimulation in situ muscle segments near the motor endplates elongate while segments away from the endplates shorten. We show that both spatial and temporal inhomogeneities in muscle deformation occurred during contraction. Muscle plateau shortening strain equalled 4.1%. Maximal plateau shortening of a muscle segment was much larger (9.6%) and occurred distally (at 0.26 of the scaled length of the muscle). Manipulating torque levels by decreasing the stimulation frequency at the same muscle length induced a decrease in torque ( approximately 20%) with a smaller effect on the level and no effect on the pattern of muscle deformation. During relaxation, distal segments actively shortened at the expense of proximal muscle segments, which elongated. The segments undergoing lengthening were nearer to motor endplates than segments undergoing shortening. In conclusion, the present study provides experimental data on magnitude of contraction-induced deformation needed for validation of numerical models. Local muscle deformation is heterogeneous both temporally and spatially and may be related to proximity to the motor endplates.

Strain distribution on rat medial gastrocnemius (MG) during passive stretch.

Deformation of the surface of passive medial gastrocnemius muscle (MG) was measured in vivo while performing a hysteresis test. The gastrocnemius muscle of male rats were dissected free and the distal tendon was cut. The lateral head was separated from the medial head. The muscle origins were left intact. 60-70 fluorescent, polystyrene spheres (diameter 0.7 mm) were attached to the surface of the MG. During the experiment, two-dimensional video recordings of the movements of the MG were made. The coordinates of the marker centroids were obtained by computer processing of digitized images and marker displacements as a function of time were calculated. Green-Lagrange strains in two principal directions were calculated (epsilon 1, epsilon 2) for three specimens. epsilon 1 had approximately the same direction as the muscle fibers. The longitudinal strain of the fibers (20-30%) was larger than the strain of the aponeurosis (1-5%); p < 0.001. No significant difference was found between the values of the transverse strains of muscle fibers and aponeurosis; the value of epsilon 2 was -6 to -9% for both tissue structures.

Mechanical blood-tissue interaction in contracting muscles: a model study.

A finite element (FE) model of blood perfused biological tissue has been developed. Blood perfusion is described by fluid flow through a series of 5 intercommunicating vascular compartments that are embedded in the tissue. Each compartment is characterized by a blood flow permeability tensor, blood volume fraction and vessel compliance. Local non-linear relationships between intra-extra vascular pressure difference and blood volume fraction, and between blood volume fraction and the permeability tensor, are included in the FE model. To test the implementation of these non-linear relations, FE results of blood perfusion in a piece of tissue that is subject to increased intramuscular pressure, are compared to results that are calculated with a lumped parameter (LP) model of blood perfusion. FE simulation of blood flow through a contracting rat calf muscle is performed. The FE model used in this simulation contains a transversely isotropic, non-linearly elastic description of deforming muscle tissue, in which local contraction stress is prescribed as a function of time. FE results of muscle tension, total arterial inflow and total venous outflow of the muscle during contraction, correspond to experimental results of an isometrically and tetanically contracting rat calf muscle.

Passive transverse mechanical properties of skeletal muscle under in vivo compression.

The objective of the present study is to determine the passive transverse mechanical properties of skeletal muscle. Compression experiments were performed on four rat tibialis anterior muscles. To assess the stress- and strain-distributions in the muscle during the experiment, a plane stress model of the cross section was developed for each muscle. The incompressible viscoelastic Ogden model was used to describe the passive muscle behaviour. The four material parameters were determined by fitting calculated indentation forces on measured indentation forces. The elastic parameters, mu and alpha, were 15.6+/-5.4 kPa and 21.4+/-5.7, respectively. The viscoelastic parameters, delta and tau, were 0.549+/-0.056 and 6.01+/-0.42 s. When applying the estimated material parameters in a three-dimensional finite element model, the measured behaviour can be accurately simulated.

Nonhomogeneous permeability of canine anulus fibrosus.

STUDY DESIGN: This report examines the permeability coefficient and aggregate modulus of slices of anulus cut from canine lumbar intervertebral discs. OBJECTIVES: To examine the influence of radial position on the properties of these materials, including outer samples with intact anulus edge. SUMMARY OF BACKGROUND DATA: The outer edge of anulus fibrosus shows radial bulge during axial compression of motion segments. The radial bulge increases monotonically when the axial compression is sustained for several hours, until a plateau is reached. Triphasic modeling of axial compression shows that this time course of radial bulge can not be obtained using a uniform permeability coefficient according to values in the literature. METHODS: Confined consolidation experiments (controlled load) were designed to measure the time course of uniaxial deformation of samples of anulus that were 4 mm in diameter and 1 mm tall. The rotation symmetry axis of the samples was defined in the radial direction of the disc. The radial permeability coefficient and the aggregate modulus were determined using the consolidation data and the linear biphasic theory. RESULTS: The permeability coefficient was lower at the periphery than in deeper layers of the anulus. Outer samples with outer surfaces that were 0.0-0.5 mm from the anulus edge had an average permeability coefficient of (1.02 +/- 0.57) x 10(-16) m4/Ns (n = 24). Inner samples that were 2.0-2.5 mm from the anulus edge had an average permeability coefficient of (2.81 +/- 0.98) x 10(-16) m4/Ns (n = 13). The aggregate modulus HA of outer samples was significantly higher (HA = 1.56 +/- 0.34 MPa) than that of inner samples (HA = 1.31 +/- 0.47 MPa). CONCLUSIONS: The fact that the outer anulus is less permeable than the inner anulus may explain why radial bulge of anulus fibrosus increases monotonically in time to an equilibrium value during sustained axial compression of a motion segment.

Ankle-foot orthosis has limited effect on walking test parameters among patients with peripheral ankle dorsiflexor paresis.

The aim of this study was to determine the relationship between ankle dorsiflexor strength and performances on several walking tests and to determine the effect of ankle-foot orthosis (AFO) use on walking tests. The following tests were used: 10-metre walking test (with and without three stairs), a complex walking task (6-minute walk with cognitive loading) and a subjective evaluation (SIP68 mobility scale and questionnaire). Isometric strength of the ankle dorsiflexors was measured. All walking tests were performed with and without AFO in random order. When relating torque values to walking performances, the highest correlation was found with the "10 metre" and "10 metre with stairs" test (r = -0.51, i.e. an inverse relationship). No threshold in the degree of paresis was found below which walking disability suddenly increased. No significant improvement could be demonstrated from AFO use on the 10-metre tests. Improvement on the 6-minute test was nearly significant (p = 0.06), the questionnaire revealed a positive opinion on AFO use related to overall walking function and effort. Thus, we have to conclude that these walking tests do not aid the clinician in estimating the severity of (progression of) the paresis nor to detect differences in degree of paresis between subjects.

An ionised/non-ionised dual porosity model of intervertebral disc tissue.

The volume of the intrafibrillar water space--i.e. the water contained inside the collagen fibres--is a key parameter that is relevant to concepts of connective tissue structure and function. Confined compression and swelling experiments on annulus fibrosus samples are interpreted in terms of a dual porosity model that distinguishes between a non-ionised intrafibrillar porosity and an ionised extrafibrillar porosity. Both porosities intercommunicate and are saturated with a monovalent ionic solution, i.c. NaCl. The extrafibrillar fixed charge density of the samples is assessed using radiotracer techniques and the collagen content is evaluated by measurement of hydroxyproline concentration. The interpretation of the experimental data yields values for the intrafibrillar water content, the average activity coefficient of the ions, the Donnan osmotic coefficient, the fraction of intrafibrillar water, the stress-free deformation state, and an effective stress-strain relationship as a function of the radial position in the disc. A linear fit between the second Piola-Kirchhoff effective stress and Green-Lagrange strain yielded an effective stiffness: H(e)=1.087 +/- 0.657 MPa. The average fraction of intrafibrillar water was 1.16 g/g collagen. The results were sensitive to changes in the activity and osmotic coefficients and the fraction of intrafibrillar water. The fixed charge density increased with distance from the outer edge of the annulus, whereas the hydroxyproline decreased.

Value of biomechanical macromodels as suitable tools for the prevention of work-related low back problems.

Biomechanical macromodels are evaluated with respect to their possible usefulness for health professionals and ergonomists, as well as for applied research on the prevention of low back problems. It is concluded that in the context stated geometrically simple models, in particular the model by Schultz and co-workers, are to be favoured over more complex models. However, load predictions in extreme trunk postures should be dealt with carefully. It is recommended that the model load predictions should be used only in the comparison of work situations and not for an assessment of the absolute acceptability of a work situation. Low back problems are related to mechanical (over)load at work. This study shows the pros and cons of various biomechanical macromodels as tools for health professionals and ergonomists, as well as for applied research on the prevention of work-related low back problems.

Uniaxial tensile testing of canine annulus fibrosus tissue under changing salt concentrations.

Recent modelling efforts in the field of mechanics of the intervertebral disc, demonstrate that the deformation properties of intervertebral disc tissue are intimately linked to compositional changes. This paper presents uniaxial tensile relaxation experiments of canine annulus fibrosus tissue under stepwise changes of external salt concentration.

Three dimensional registration of mechanical bladder activity using polystyrene fluorescent spheres: A technical note.

Optical marker tracing methods have been applied successfully in recent years to quantify local material deformation of heart tissue, skin and striated muscles. In this study, polystyrene fluorescent spheres (d = 0.6 mm) are glued to the ventral serosal bladder wall in the rabbit. Three dimensional video registration of the polystyrene spheres is used to calculate two directions of principal strain (epsilon (1), epsilon (2) ) on the bladder surface in vivo. The aim is to investigate the feasibility of the technique for this new application in two experimental circumstances: during spontaneous bladder wall activity and after electrical stimulation of bladder innervating nerve fibers. During spontaneous activity, random contraction and relaxation occurred simultaneously and separately across the bladder wall for the two principal strains epsilon (1) and epsilon (2). After extradural electrical stimulation of sacral nerve root S2, the principal strains epsilon ( 1) and epsilon (2) synchronized in time in such a way that epsilon ( 1) and epsilon (2) both represented contraction or both represented relaxation. One and the same bladder wall area passed through phases of contraction followed by relaxation and vice versa. After multiple stimulation periods, the coordination between the two principal strains during stimulation was reduced. This technique allows to identify local areas of contraction and relaxation in the intact bladder wall in vivo. Three dimensional video registration of polystyrene fluorescent spheres to study bladder wall contraction and its relaxation proved to be a feasible technique, with which electrical stimulation effects and spontaneous activity could be measured.

Determination of muscle fibre orientation using Diffusion-Weighted MRI.

Biomechanical studies have shown that the distribution of stress and strain in biological tissue is strongly dependent on fibre orientation. Therefore, to analyze the local mechanical load, accurate data on muscle fibre orientation are needed. Traditional techniques to determine fibre orientation are inherently invasive. Here we used Diffusion Weighted MRI to non-invasively determine, in each image voxel of 0.23 x 0.23 mm, the diffusion tensor of water in the cat semimembranosus muscle. The direction corresponding to the largest eigenvector of this tensor was calculated. This direction was found to correspond qualitatively to the muscular fibre direction, as determined by visual inspection.

Three-dimensional reconstruction of the rat triceps surae muscle and finite element mesh generation of the gastrocnemius medialis muscle.

In order to simulate blood flow in skeletal muscle, our group has developed a finite element description of perfused skeletal muscle. This model requires input parameters concerning the vascular system, muscle contraction and the geometry of a muscle, including its aponeuroses. The objective of the present paper is to create a geometrical reconstruction of the rat gastrocnemius medialis muscle that can be incorporated in the finite element model. Since this muscle is connected to the plantaris and the gastrocnemius lateralis muscle, a detailed computer graphical reconstruction of the triceps surae muscle, based on histological cross-sections, has been accomplished first. Using this reconstruction, relevant sections were selected to create the finite element mesh of the gastrocnemius medialis muscle. Special attention was payed to the location of the aponeuroses. The mesh can be used in finite element simulations of perfused skeletal muscle.

Skeletal Muscle wasting and contractile performance in septic rats.

We investigated the temporal effects of sepsis on muscle wasting and function in order to study the contribution of wasting to the decline in muscle function; we also studied the fiber-type specificity of this muscle wasting. Sepsis was induced by injecting rats intraperitoneally with a zymosan suspension. At 2 h and at 2, 6, and 11 days after injection, muscle function was measured using in situ electrical stimulation, Zymosan injection induced severe muscle wasting compared to pair-fed and ad libitum fed controls. At 6 days, isometric force-generating capacity was drastically reduced in zymosan-treated rats. We conclude that this was fully accounted fo by the reduction of muscle mas. At day 6, we also observed increased activity of the 20S proteasome in gastrocnemius but not soleus muscle from septic rats. In tibialis anterior but not in soleus, muscle wasting occurred in a fiber-type specific fashion, i.e., the reduction in cross-sectional area was significantly smaller in type 1 than type 2A and 2B/X fibers. These findings suggest that both the inherent function of a muscle and the muscle fiber-type distribution affect the responsiveness to catabolic signals.