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A finite element model for describing the effect of muscle shortening on surface EMG.

A finite-element model for the generation of single fiber action potentials in a muscle undergoing various degrees of fiber shortening is developed. The muscle is assumed fusiform with muscle fibers following a curvilinear path described by a Gaussian function. Different degrees of fiber shortening are simulated by changing the parameters of the fiber path and maintaining the volume of the muscle constant. The conductivity tensor is adapted to the muscle fiber orientation. In each point of the volume conductor, the conductivity of the muscle tissue in the direction of the fiber is larger than that in the transversal direction. Thus, the conductivity tensor changes point-by-point with fiber shortening, adapting to the fiber paths. An analytical derivation of the conductivity tensor is provided. The volume conductor is then studied with a finite-element approach using the analytically derived conductivity tensor. Representative simulations of single fiber action potentials with the muscle at different degrees of shortening are presented. It is shown that the geometrical changes in the muscle, which imply changes in the conductivity tensor, determine important variations in action potential shape, thus affecting its amplitude and frequency content. The model provides a new tool for interpreting surface EMG signal features with changes in muscle geometry, as it happens during dynamic contractions.

Action Potentials↗

"RED" SKELETAL MUSCLE FIBERS: RELATIVE INDEPENDENCE OF NEURAL CONTROL.

Studies on skeletal muscle of rats and mice indicate significant differences between the behaviors of the two basic types of muscle fibers during the development of denervation atrophy. While the diameter of fibers possessing sarcoplasmic granularity ("red" fibers) is preserved for a long time after complete severance of motor and sensory nerves, fiber with agranular cytoplasm ("white" fibers) undergo rapid atrophy. It is concluded that, at least in this respect, the "white" fibers are more dependent on neural control than are the "red" fibers.

Animals↗

Selective activation of quadriceps muscle fibers according to bicycling rate.

Moving average electromyography (MA) of quadriceps muscle bellies has been recorded during bicycling at different rates (30-70 cycles/min) or forces (1-3 kg). For power increments (50-100%) achieved by increasing force at constant rate, MA during pedal downstroke always increased. For similar power increments achieved by increasing the rate at constant force, MA did not increase (37% of cases), increased less (37%), or increased similarly (26%). Investigations by others on the rat suggest that the lack of increase of MA despite power increment was not compensated by other muscle activity; hence it indicates a shift from slow to fast fibers, which provide greater power per unit stimulus. Smaller increase of MA with increasing rate rather than force at isopower could depend on this shift or on muscle properties, if operating on ascending limb of power-velocity curve. This, however, does not seem the case for slow fibers, which should develop peak power at about 25 cycles/min. Hence, fibers of quadriceps muscle of humans seem selectively activated according to movement speed, as previously found in inspiratory muscles of rabbits.

Adult↗

Experience from a long-term carcinogenicity study with intraperitoneal injection of biosoluble synthetic mineral fibers.

The carcinogenic potential in the intraperitoneal cavity of three newly developed biosoluble insulation glass wool fibers (M, P, and V) and one newly developed biosoluble insulation stone wool fiber (O) was investigated and compared to that of a previously developed soluble glass fiber (B). The in vitro dissolution coefficient of the three glass wool fibers ranged from 450 to 1037 ng/cm(2) x h and was 523 ng/cm(2) x h for the stone wool fiber. The in vitro dissolution coefficient of the B fiber was 580 ng/cm(2) x h. Groups of female Wistar rats (strain Crl: Wi BR) were exposed by repeated injections to doses of 0.5, 2, and 5 x 10(9) WHO fibers, which corresponds to between 41 mg to 724 mg fiber injected. In addition, 2 groups of crocidolite were used as positive controls at doses of 0.1 x 10(9) and 1 x 10(9) WHO fibers (0.5 and 5 mg). The in vitro dissolution coefficient of crocidolite is estimated to be approximately 1 ng/cm(2) x h. The protocol of the study and the size distribution of the test samples conformed to the European Commission Protocol EUR 18748 EN, and the study was executed under Good Laboratory Practice conditions. Two of the new insulation wools, fibers M and 0, showed no statistically significant tumorigenic response even at the very high dose of 5 x 10(9) WHO fibers injected. Fibers P and V showed a small tumorigenic response in the ip cavity similar in magnitude to the B fiber, which has been declared in the German fiber regulations as a noncarcinogenic fiber. The response to the soluble insulation fibers was notably different from that of the known carcinogen crocidolite, which produced 53% tumors at a comparatively low dose of 0.1 x 10(9) WHO fibers. The incidence of mesothelioma was found to be highly correlated to the incidence of intra-abdominal nodules and masses at different sites. The incidence of abdominal nodules and masses was highly correlated to the number of animals with ascites. The incidence of chronic peritonitis with fibrotic nodules at different organs also correlated with the incidence of mesotheliomas. Differences in etiology were observed between the massive doses of the highly soluble insulation wools when injected directly into the ip cavity and the lower doses of the extremely insoluble fiber crocidolite. The variability in this reaction and the impairment of animal health put into question the value of these massive doses in evaluating the carcinogenic response of soluble insulation wools. All of the fibers tested fulfilled the exoneration criteria with respect to carcinogenicity according to the European Directive 97/69/EC ("an appropriate intra-peritoneal test has not expressed signs of excessive carcinogenicity"). The dose as defined in the EC-Protocol EUR 18748 EN was 1 x 10(9) WHO fibers with a defined geometric spectrum. The influence of fiber dimensions on the ip tumor response and the difficulty in assessing the influence of the difference in background levels between this and previous studies make direct application of the German TRGS 905 criteria difficult; however, by comparison to fiber B, which in the TRGS 905 is considered as a noncarcinogenic fiber, all of the synthetic mineral fiber types tested in this study also appear to meet the intended German criteria for exoneration.

Abdominal Neoplasms↗

[Developments in preparation and experimental method of solid phase microextraction fibers].

Solid phase microextraction (SPME) is a simple and effective adsorption and desorption technique, which concentrates volatile or nonvolatile compounds from liquid samples or headspace of samples. SPME is compatible with analyte separation and detection by gas chromatography, high performance liquid chromatography, and other instrumental methods. It can provide many advantages, such as wide linear scale, low solvent and sample consumption, short analytical times, low detection limits, simple apparatus, and so on. The theory of SPME is introduced, which includes equilibrium theory and non-equilibrium theory. The novel development of fiber preparation methods and relative experimental techniques are discussed. In addition to commercial fiber preparation, different newly developed fabrication techniques, such as sol-gel, electronic deposition, carbon-base adsorption, high-temperature epoxy immobilization, are presented. Effects of extraction modes, selection of fiber coating, optimization of operating conditions, method sensitivity and precision, and systematical automation, are taken into considerations in the analytical process of SPME. A simple perspective of SPME is proposed at last.

Chromatography, Gas↗

A nuclear magnetic resonance technique for determining hybridoma cell concentration in hollow fiber bioreactors.

We have developed a technique for determining cell concentration in a hollow fiber bioreactor based on 23Na nuclear magnetic resonance (NMR) spectroscopy. Cell concentrations determined with this method agreed closely with concentrations calculated from 31P NMR nucleoside triphosphate (NTP) measurements and oxygen consumption rate measurements. Oxygen transfer limitations, which can complicate cell mass determinations based on oxygen consumption rates, were shown to be negligible for the bioreactor used. Specific antibody production rates in hollow fiber culture, calculated from these cell number estimates, were similar to those found in suspension culture for this cell line.

Animals↗

Structure and development of central arborizations of hair follicle primary afferent fibers.

The present study describes the structure and development of the flame-shaped central arborizations of hair follicle (HF) afferents in the lumbosacral spinal cord of the rat. Tissue was processed according to the rapid Golgi method at successive stages of development from embryonic day 17 through postnatal day 30. Collaterals of most HF afferents were found to enter the dorsal horn via a characteristic U-shaped pathway which often parallels the vascular pattern. The HF collaterals can first be identified at embryonic day 19 and by postnatal day 5 they have established the dorso-ventral and rostro-caudal limits of their field of arborization. Dorsally the arbors extend no further than the inner zone of lamina II (IIi) at any stage of development. Short aberrant branches were observed on some HF collaterals during the prenatal period but none of these developed synaptic terminals or contributed branches to other arbors. Each HF collateral formed a single well defined flame-shaped arbor with a distribution and branching pattern which could be distinguished from that of other afferents throughout the postnatal developmental period. Two types of HF collaterals were observed. Structure and distribution patterns suggest that type I collaterals are derived from G and T HF afferents while collaterals in the type II category are probably derived from both G and T as well as D (A-delta) HF afferents. Type I collaterals divide into well defined medial and lateral collateral branches which arborize mainly in lamina III with a few branches to lamina IV. Type II collaterals are characterized by a delicate arbor which is more vertically oriented than type I arbors.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Fiber type composition of the plantarflexors of giraffes (Giraffa camelopardalis) at different postnatal stages of development.

1. A sample of fibers from deep (close to the bone) and superficial (away from the bone) regions of the plantaris (PLT) and medial (MG) and lateral (LG) gastrocnemius muscles of a neonatal, a 17-day-old and an adult giraffe were typed qualitatively as dark or light based on alkaline preincubation myosin ATPase staining properties and then sized. 2. Each muscle at all ages showed a higher percentage and a larger cross-sectional area (CSA) or light ATPase fibers in the deep than the superficial region. This relationship was qualitatively, although not quantitatively, similar to that reported in hindlimb muscles of other mammals. 3. At all ages, the PLT, the deepest muscle in the synergistic group, had the highest relative total CSA of light ATPase fibers among the muscles sampled. 4. At birth, the PLT had an unusually high percentage of light ATPase fibers in comparison to that found in the same muscle of other mammals. With age, the total CSA of light ATPase fibers increased dramatically in the PLT and decreased slightly in the MG and LG. 5. These data suggest that the PLT, especially the deep portion, may functionally replace the soleus muscle which is absent in the giraffe. In addition, the fiber type results demonstrate that the changes in the fiber type composition of individual muscles observed at different postnatal ages in the giraffe are relatively similar to that reported in smaller mammals, suggesting the existence of similar regulatory mechanisms.

Age Factors↗

Alteration in synaptic inputs through C-afferent fibers to substantia gelatinosa neurons of the rat spinal dorsal horn during postnatal development.

The change in synaptic inputs through primary afferent C- and A-fibers during postnatal development was examined in substantia gelatinosa neurons of a rat spinal cord slice with an attached L5 dorsal root by use of the blind whole-cell patch-clamp technique; the synaptic responses were compared between the slices obtained from immature (postnatal days 21-23) and mature (postnatal days 56-60) male rats. The mono- and/or polysynaptic afferent inputs were monitored by recording glutamatergic excitatory postsynaptic currents and potentials evoked by stimulating C- and A-fibers, the identification of which was based on the values of threshold stimulus intensity and of the conduction velocity of the fibers, determined by intracellular recordings from dorsal root ganglion neurons. Immature substantia gelatinosa neurons received synaptic inputs through Abeta-, Adelta- and C-afferents, with proportions of 51%, 46% and 36%, respectively. In mature substantia gelatinosa neurons, C- and Adelta-afferent inputs were increased in number (to 84% and 86%, respectively), while Abeta-inputs were decreased to 9%. In both immature and mature rats, repetitive stimulation of C-afferents did not elicit any slow responses, which are longer in duration than the monosynaptic excitatory postsynaptic currents, although C-fibers are known to contain not only excitatory amino acids, but also neuropeptides such as substance P, which is thought to be involved in the production of slow responses. These results indicate that both C- and Adelta-afferents innervating substantia gelatinosa neurons are reorganized following maturation, accompanied by a withdrawal or elimination of Abeta-fibers from the substantia gelatinosa, probably due to a competition among the fibers during development. In spite of the developmental increase in C-fiber inputs, mature as well as immature substantia gelatinosa neurons did not display any slow synaptic responses, which appear to be mediated by transmitters other than excitatory amino acids.

Action Potentials↗

Localization of FMRFamide-like immunoreactivity in the brain of the viviparous skink (Chalcides chalcides).

Neuroanatomical distribution of FMRFamide-like immunoreactivity was investigated in the brain and olfactory system of the viviparous skink, Chalcides chalcides. In the adult brain FMRFamide immunoreactive (ir) perikarya were observed in the diagonal band of Broca, medial septal nucleus, accumbens nucleus, bed nucleus of the anterior commissure, periventricular hypothalamic nucleus, lateral forebrain bundle, and lateral preoptic, subcommissural, suprachiasmatic and lateral hypothalamic areas. This pattern was seen in both male and female brains. Though all major brain areas showed FMRFamide-ir innervation, the densest ir fiber network was observed in the hypothalamus. During development, ir elements were observed for the first time in embryos at mid-pregnancy. FMRFamide perikarya were located along the ventral surface of the vomeronasal nerve, in the olfactory peduncle mediobasally, as well as in the anterior olfactory nucleus and olfactory tubercle. Furthermore, some ir neurons were observed in the rhombencephalic reticular substance; however, the ir fiber network was poorly developed. Later in development FMRFamide-ir neurons appeared also in the bed nucleus of the anterior commissure as well as the rhombencephalic nucleus of solitary tract and the dorsal motor nucleus of vagus nerve. In juveniles, the distribution profile of FMRFamide immunoreactivity was substantially similar to that of the adults, with a less widespread neuronal distribution and a more developed fiber network. Ontogenetic presence of FMRFamide immunoreactivity in the nasal area has been linked to the presence of a nervus terminalis in this reptile.

Animals↗

Hippocampal mossy fiber sprouting and synapse formation after status epilepticus in rats: visualization after retrograde transport of biocytin.

In complex partial epilepsy and in animal models of epilepsy, hippocampal mossy fibers appear to develop recurrent collaterals that invade the dentate molecular layer. Mossy fiber collaterals have been proposed to subserve recurrent excitation by forming granule cell-granule cell synapses. This hypothesis was tested by visualizing dentate granule cells and their mossy fibers after terminal uptake and retrograde transport of biocytin. Labeling studies were performed with transverse slices of the caudal rat hippocampal formation prepared 2.6-70.0 weeks after pilocarpine-induced or kainic acid-induced status epilepticus. Light microscopy demonstrated the progressive growth of recurrent mossy fibers into the molecular layer; the densest innervation was observed in slices from pilocarpine-treated rats that had survived 10 weeks or longer after status epilepticus. Thin mossy fiber collaterals originated predominantly from deep within the hilar region, crossed the granule cell body layer, and formed an axonal plexus oriented parallel to the cell body layer within the inner one-third of the molecular layer. When sprouting was most robust, some recurrent mossy fibers at the apex of the dentate gyrus reached the outer two-thirds of the molecular layer. The distribution and density of mossy fiber-like Timm staining correlated with the biocytin labeling. When viewed with the electron microscope, the inner one-third of the dentate molecular layer contained numerous mossy fiber boutons. In some instances, biocytin-labeled mossy fiber boutons were engaged in synaptic contact with biocytin-labeled granule cell dendrites. Granule cell dendrites did not develop large complex spines ("thorny excrescences") at the site of synapse formation, and they did not appear to have been permanently damaged by seizure activity. These results establish the validity of Timm staining as a marker for mossy fiber sprouting and support the view that status epilepticus provokes the formation of a novel recurrent excitatory circuit in the dentate gyrus. Retrograde labeling with biocytin showed that the recurrent mossy fiber projection often occupies a considerably greater fraction of the dendritic region than previous studies had suggested.

Animals↗

Combined imaging and chemical sensing using a single optical imaging fiber.

Despite many innovations and developments in the field of fiber-optic chemical sensors, optical fibers have not been employed to both view a sample and concurrently detect an analyte of interest. While chemical sensors employing a single optical fiber or a noncoherent fiberoptic bundle have been applied to a wide variety of analytical determinations, they cannot be used for imaging. Similarly, coherent imaging fibers have been employed only for their originally intended purpose, image transmission. We herein report a new technique for viewing a sample and measuring surface chemical concentrations that employs a coherent imaging fiber. The method is based on the deposition of a thin, analyte-sensitive polymer layer on the distal surface of a 350-microns-diameter imaging fiber. We present results from a pH sensor array and an acetylcholine biosensor array, each of which contains approximately 6000 optical sensors. The acetylcholine biosensor has a detection limit of 35 microM and a fast (< 1 s) response time. In association with an epifluorescence microscope and a charge-coupled device, these modified imaging fibers can display visual information of a remote sample with 4-microns spatial resolution, allowing for alternating acquisition of both chemical analysis and visual histology.

Acetylcholinesterase↗

Effect of alfalfa fiber substrate on culture counts of rumen bacteria.

A medium has been developed using alfalfa fiber as the sole substrate. It gave high culture counts (3 X 10(9) to 8 X 10(9)/ml) of rumen bacteria. When this medium was combined with the medium 98-5 of Bryant and Robinson, modified to contain 33% rumen fluid instead of 40% clarified rumen fluid, a higher count was obtained than with either medium alone.

Anaerobiosis↗

Ophthalmoscopic model of nerve fiber bundle defects in monkey eyes produced by intraocular retinal coagulation.

The development of nerve fiber bundle defects in monkey eyes by means of intraocular retinal coagulation, such as endodiathermy, intraocular cryotherapy or endophotocoagulation was confirmed by ophthalmoscopy. Two to 4 days after the application of intraocular retinal coagulation, the findings could be already observed. Endodiathermy especially could provide the typical ophthalmoscopic findings of nerve fiber bundle defects without any abnormal fluorescein angiographic findings, while endophotocoagulation and intraocular cryotherapy induced hyperfluorescence due to window defect at the coagulation site. Therefore, endodiathermy in monkey eyes can provide an excellent ophthalmoscopic model of the nerve fiber bundle defects. Histological findings 8 weeks after the coagulation showed thinning of the retinal nerve fiber layer and gliosis occurring in the retinal portion corresponding to the lesion.

Animals↗

Muscle fiber regeneration in nerve-intact and free skeletal muscle autografts in cats.

Extensor digitorum longus (EDL) muscles of adult cats were transplanted as free autografts (FRA) with the nerve severed or as nerve-intact autografts (NIA) with the nerve retained. Histochemical and contractile properties of NIA and FRA were analyzed at selected times from 1 to 14 wk after surgery. Regeneration was qualitatively similar in NIA and FRA. Regenerating fibers were observed in both NIA and FRA within 2 wk. After 14 wk there were fewer type I fibers in both NIA and FRA than in control EDL muscles. Capillarity was greater in NIA than FRA, but both types of autografts had significantly reduced capillarity relative to control muscles. Mean fiber area, muscle mass, and absolute tension development were greater in NIA than FRA but did not reach control muscle values. Muscle mass, mean fiber area, and contractile properties, but not the proportion of type I fibers, develop toward control values more quickly in autografts with the nerve left intact.

Animals↗

Immunohistochemical and ultrastructural localization of CGRP-positive nerve fibers at the epiphyseal trabecules facing the growth plate of rat femurs.

We performed immunohistochemical and ultrastructural studies to disclose a possible relationship between nerve fibers and bone metabolism. Immunohistochemical distribution of calcitonin gene-related peptide (CGRP)-positive nerve fibers during bone development was assessed in the femurs of rats. CGRP-positive nerve fibers were denser in the epiphysis than in the metaphysis. These nerve fibers particularly ran along the epiphyseal trabecules facing the growth plate and came in contact with osteoclasts. Many osteoclasts at the epiphyseal trabecules facing the growth plate contained abundant toluidine blue and periodic acid-Schiff-positive granules. Electron microscopy revealed that these osteoclasts have many membrane-bound, electron-dense granular structures and dilated cisterns of rough endoplasmic reticulum containing electron-dense material. They were often surrounded by clear cells displaying features of nerve fiber and had no ruffled border. Furthermore, ultrastructural observations revealed electron-dense structures coating the cytoplasmic side of plasma membranes of the nerve fibers. We also observed coated pits in the cytoplasm of the osteoclasts facing the nerve fibers. To further clarify the role of innervation, we compared trabecules of rats undergoing denervation of the sciatic nerve with those from unoperated rats. Denervation resulted in a significant increase in the number of cement lines on the epiphyseal trabecules facing the growth plate. These results suggest that the osteoclasts at the epiphyseal trabecules facing the growth plate are in part regulated by CGRP-positive nerve fibers. Thus, CGRP-positive nerve fibers could be a crucial element in bone metabolism during bone growth and development.

Animals↗

Phosphorylating pathways and fatigue development in contracting Xenopus single skeletal muscle fibers.

To investigate the differential contribution of oxidative and substrate-level phosphorylation to force production during repetitive, maximal tetanic contractions, single skeletal muscle fiber performance was examined under conditions of high-oxygen availability and anoxia. Tetanic force development (P) was measured in isolated, single type-1 muscle fibers (fast twitch; n = 6) dissected from Xenopus lumbrical muscle while being stimulated at increasing frequencies (0.25, 0.33, and 0.5 Hz), with each frequency lasting 2 min. Two separate work bouts were conducted, with the perfusate PO(2) being either 0 or 159 mmHg. No significant (P < 0. 05) difference was found in the initial peak tensions (P(0)) between the high (334 +/- 57 kPa) and the low (325 +/- 41 kPa) PO(2) treatment. No significant difference in P was observed between the treatments during the first 50 s. However, a significant difference in force production was observed between the high (P/P(0) = 0.96 +/- 0.02) and the low PO(2) condition (P/P(0) = 0.92 +/- 0.02) by 60 s of work. After 60 s, steady-state force production was maintained during the high compared with the low PO(2) condition until stimulation frequency was increased, at which point developed tension during the high PO(2) condition began to decline. Time to fatigue (P/P(0) = 0.3) was reached significantly sooner during the low (250 +/- 16 s) than the high PO(2) condition (367 +/- 28 s). These results demonstrate that during the first 50 s of 0.25-Hz contractions, substrate-level phosphorylation has the capacity to maintain force and ATP hydrolysis when oxidative phosphorylation is absent. This period was followed by an oxygen-dependent phase in which force generation was maintained during the high PO(2) condition (but not during the low PO(2) condition) until the onset of a final fatiguing phase, at which a calculated maximal rate of oxidative phosphorylation was reached.

Adenosine Triphosphatases↗