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Biomedical subjects

W G Hopkins

Publications and source records attributed to W G Hopkins.

At least 73 records · Page 4Linked to original sources

The meaning of "physical fitness".

The understanding of the term "physical fitness" was determined for a randomly selected sample (n = 94) of a population using a self-administered mailed questionnaire. Subjects were asked to state and give a reason for their perceived level of physical fitness, to state their perceived performance level in a number of physical fitness tests (muscular strength, daily physical work capacity, fatness, level of regular physical exercise, exercise speed, and body flexibility), and to rate how well these tests measure physical fitness. The reason most frequently stated for perceived level of physical fitness was the level of habitual physical activity (43%); significantly less frequently (P less than 0.01-0.0001) cited were reasons related to health (23%), physical performance (12%), and obesity (3%). The variation in perceived level of physical fitness was best explained by the variation in imagined regular exercise and fatness (r2 = 0.66, P less than 0.0001) with no significant additional contribution from imagined performance in remaining fitness tests. The measurement of regular exercise was most favored as a test of physical fitness. These results, taken together with evidence of the physical and psychological health benefits of regular exercise, imply that the most appropriate measure of physical fitness for the average person is an assessment of the habitual physical activity level.

Attitude to Health↗

Physical fitness of blind and sighted children.

Twenty-seven children (age 7-17 years) with varying degrees of blindness but with no other known disorder were assessed for physical fitness. Twenty-seven randomly selected children with normal eyesight were also assessed. Maximum oxygen uptake (VO2max) was measured directly during a progressive exercise test on a treadmill. There was a significant and substantial reduction in VO2max in totally blind children (mean +/- standard deviation 35.0 +/- 7.5 ml X min-1 X kg-1) compared with normal children (45.9 +/- 6.6 ml X min-1 X kg-1). Partially sighted children had a significant but smaller reduction in VO2max. Fitness assessed by a step-test was significantly reduced in the visually impaired children, and skin-fold thickness was also significantly greater in totally blind children. The level of habitual physical activity for each child, as assessed by a questionnaire, correlated with VO2max (r = 0.53, p less than 0.0001). Blind children were significantly less active than normal children, and the difference between mean VO2max for blind and normal children became non-significant when their different activity levels were taken into account. It is concluded that totally blind children are less fit than other children at least partly because of their lower level of habitual activity.

Adolescent↗

End-plate growth exceeds nerve terminal growth in juvenile mice.

An electron microscopic investigation of neuromuscular synapses of the tensor fasciae latae muscle in juvenile mice was carried out to determine the relationship between the presynaptic (nerve terminal) and postsynaptic (end-plate plaque) elements during the major phase of postnatal growth. In the first month of life growth of the synapse was accompanied by a decrease in the size of pre- and postsynaptic elements in cross section. A decrease in the fraction of the width of end-plate plaque opposed by nerve terminal was also observed to age 2 months, and in older animal some end-plate plaques were completely unopposed by nerve terminal. This implies that end plates grow at a faster rate than nerve terminals and suggests that the availability of uninnervated, differentiated end plate permits or promotes nerve terminal growth.

Aging↗

Isolation and in vitro translation of polysomes from mature rye leaves.

Cytoplasmic polysomes have been prepared from mature leaves of winter rye (Secale cereale L. cv Puma). This is the first time a method has been developed for isolation of highly polymerized polysomes from mature leaves. The degree of intactness of isolated plant polysomes has been determined by two independent but complementary methods: size class distribution by sucrose gradient centrifugation and in vitro translation. The polymerization of isolated polysomes was estimated by the ratio of the proportion of large polysomes to the proportion of small polysomes obtained from the profiles. Our results show that the composition of the optimal polysome isolation buffer for mature rye leaves is different from that reported for young tobacco and pea leaves. Polysomes were translated in vitro with the S-105 wheat germ fraction. The degree of polysome polymerization has a significant effect on their in vitro translation since both the incorporation of amino acid and the presence of high molecular weight polypeptides are proportional to the large polysomes/small polysomes ratio. This study emphasizes the need to evaluate isolation conditions carefully before proceeding with polysome studies in any particular tissue or in tissues under different physiological status.

Journal Article↗

Polysomes from winter rye seedlings grown at low temperature : I. Size class distribution, composition, and stability.

We have studied the influence of growth at low temperature on size class distribution, stability and composition of leaf cytoplasmic polysomes from rye seedlings (Secale cereale, cv Puma) grown at 5 degrees C and at 20 degrees C. Leaves of seedlings grown at 5 degrees C contain 2.7 times more cytoplasmic polysomes (expressed on a DNA basis) and the polysome size class distribution is skewed toward larger polysomes. These changes were more pronounced in the free polysome fraction than in the membrane-bound fraction. The melting point of the total ribosome fraction from cold-grown leaves was decreased by 3.7 degrees C. Electrophoresis did not reveal any difference in the rRNA or in core-ribosomal proteins (KCl nondissociable) following growth at low temperature. Some differences were noted in peripheral ribosomal proteins. This study is the first to examine the effect of growth at low and high temperatures on polysome metabolism using plants of similar developmental stage. Polysome quantity, polymerization, melting point and peripheral ribosomal proteins in rye seedlings are modified during growth at low temperature.

Journal Article↗

Effect of age and muscle type on regeneration of neuromuscular synapses in mice.

A silver stain was used to investigate the regeneration of nerve terminals in mouse diaphragm, superior gluteus and tensor fasciae latae (TFL) muscles following nerve crush injury at different ages (9-530 days). The development of myelinated terminal branches and the development and elimination of terminal sprouts were little affected by age or muscle type following reinnervation. However, multiple axonal innervation developed on up to 50% of the gluteus and TFL muscle fibres, and this was subsequently eliminated only in the youngest animals.

Aging↗

Postnatal growth of motor nerve terminals in muscles of the mouse.

A reduced silver stain was used to examine the development of complexity of motor nerve terminals in the postnatal period. Terminals in three histochemically different muscles were examined in mice aged 12 days to 3 years. The total number and total length of intraterminal axon branches increases with age, but only until animals are 3 months old. Terminals become longest and most branched in the histochemically glycolytic tensor fascia latae (TFL) muscle, shortest and least branched in the oxidative diaphragm, and intermediate in the histochemically mixed gluteus muscle. In addition, myelinated terminal branches develop in TFL and to a lesser extent in the gluteus between 1 and 3 months of age. These myelinated branches appear to be produced by nodal sprouting from the penultimate node of Ranvier of the terminal axon, and also by myelination of pre-existing terminal branches. The diameter of muscle fibres also increases until animals are 3 months old, and there is a good correlation between mean fibre diameter and either mean terminal length or mean number of terminal branches when all muscles at all ages are compared. This suggests that terminal growth could be determined by muscle fibre growth; however, within any given muscle there is little or no correlation between the diameter of a muscle fibre and either the length or number of branches of its nerve terminal, suggesting that terminal morphology is not controlled solely by muscle fibre growth. The presence of a myelinated branch in a nerve terminal is also unrelated to fibre diameter within a given muscle, but again when means are compared there are good, but significantly different correlations for the three different muscles. Thus some kind of muscle or nerve type-specific property additional to a general effect of muscle fibre size influences the development of myelinated terminal branches. Between 3 and 12 months of age terminal complexity remains constant or may decrease slightly. At 19 months or older, when mice are becoming senile, a large proportion of synapses have terminal sprouts and muscle fibres become innervated by two or more distinct axons. These changes can be attributed to the death of some motor neurons and sprouting of the remaining axons.

Aging↗

Regeneration of mouse peripheral nerves in degenerating skeletal muscle: guidance by residual muscle fibre basement membrane.

The part played by basement membrane in the guidance of peripheral nerve growth in vivo has been assessed by examining the capacity of degenerating mouse muscle to support the regeneration of the cut sciatic and saphenous nerves. Ethanol and formaldehyde-fixed gluteus maximus muscles were implanted around the contralateral cut nerves. The subsequent nerve growth into the degenerating muscle was assessed by silver staining after 3, 4 and 10 days. By 4 days, linear axonal growth was seen, parallel to the length of the muscle fibres, and coinciding with the onset of degeneration of the sarcoplasm. Transverse sections of the 10 day preparations showed that over 90% of linearly growing axons were located inside the remaining sheaths of muscle fibre basement membrane. This relationship was confirmed by electron microscopy of ruthenium red-stained preparations. Both motor and sensory axons were able to grow in this manner, for electrophysiological testing revealed the presence of motor axons from the sciatic nerve, while the saphenous nerve contains only sensory axons. Identical growth was seen at 10 days in muscles caused to degenerate by incubation in distilled water. However, linear growth did not occur in live-innervated and glutaraldehyde-fixed muscles, in which muscle fibre architecture was preserved. It is concluded that basement membrane derived from muscle can promote peripheral nerve regeneration. Furthermore, both motor and sensory axons show a strong preference for growth along its inner surface, the basal lamina.

Animals↗

Axon growth in tendon organs of botulinum-paralyzed neonatal mouse muscles.

Paralysis of neonatal mouse gluteus muscles with botulinum toxin stimulated the growth not only of motor nerve terminals but also of the terminals of the sensory axons innervating the tendon organs. However, the adult mouse tendon organ axons, unlike motor nerve terminals, did not sprout in response to paralysis.

Animals↗

Effect of nerve growth factor on intramuscular axons of neonatal mice.

Newborn mice were given daily injections of nerve growth factor over the gluteus muscle. At age 9 days muscles were removed and stained to reveal intramuscular axons. Motor axons, and sensory axons of the Golgi tendon organs and muscle spindles were unaffected by nerve growth factor, but there was prolific growth of the small diameter axons that ramify over the surface of the muscle. These axons were sensory rather than sympathetic, since the degenerated following dorsal root ganglionectomy but not following sympathetic ganglionectomy. This finding supports the concept that nerve growth factor is involved in the development of sensory innervation of muscle.

Animals↗

Muscle size and motor unit survival in mice.

The soleus muscles in neonatal and adult mice were surgically reduced in size on one side of the animal. The experimental and control muscles were excised 6-48 weeks later and the number of motor units in each muscle was estimated by stimulating the muscle nerve and counting step increments in the electromyogram recorded in vitro. Multiple innervation in individual muscle fibres was then assessed by intracellular recording and by visualization of end-plates in the light microscope with cholinesterase stain. Muscle fibres were counted in cross-sections of each muscle in the light microscope. Surgical reductions in the size of the muscle during the first 3 weeks of life produced correlated reductions in the number of motor units in the muscle. This could not be attributed to masking of motor units by multiple innervation, which was always less than 10% in these muscles. The loss of motor units was greatest following reduction in muscle size in newborn mice, whereas in 6-week-old mice there was no significant loss of motor units following the operation. Thus, survival of neonatal motor units shows an age-related dependence on the number of muscle fibres available for innervation. In control muscles there was a highly significant correlation between motor unit and muscle fibre numbers, which is consistent with the hypothesis that motor neurone survival during the embryonic period of cell death is dependent upon the number of muscle fibres available for innervation.

Age Factors↗

Sprouting of mammalian motor neurones at nodes of Ranvier: the role of the denervated motor endplate.

Treatment of the mouse gluteus maximus with the organophosphorus anticholinesterase ecothiopate, 4-aminopyridine and caffeine caused necrosis of the endplate region of the muscle fibres. When these drugs were given 12 h prior to partial denervation of the muscle there was a significant reduction in the amount of nodal sprouting seen 48 h after nerve section. No such reduction was seen if the drugs were given immediately after nerve section. In the former case both innervated and denervated fibres are necrotic; in the latter, only innervated fibres are necrotic. The amount of terminal sprouting was significantly increased in the former group, showing that the drug treatment and necrosis were not directly inhibitory to nerve growth. The reduction of nodal sprouting is therefore likely to be due to necrosis of denervated fibres; this supports the hypothesis that the denervated endplate is the source of a stimulus to nodal sprouting of motoneurones.

Animals↗

Evidence for a motor nerve growth factor.

We review the evidence that a motor nerve growth factor released from muscle has wide ranging effects on the development and maintenance of muscle innervation. The actions of this putative factor on motor neurons are analogous to the actions of the well known nerve growth factor (NGF) on sympathetic and sensory neurons.

Animals↗

Importance of pathway formation for nodal sprout production in partly denervated muscles.

Experiments were carried out to investigate possible factors controlling nodal sprout growth in partly denervated mouse gluteal muscles. Pretreatment of the muscle with botulinum toxin for up to 20 days, which produces denervation-like change and elicits terminal and nodal sprouting, increased reinnervation by terminal sprouting after partial denervation but did not alter the rate of reinnervation by nodal sprouts. This implies that nodal sprout growth is not limited by the development of an adequate growth stimulus from denervated muscle. A disto-proximal gradient of degeneration was observed in denervated intramuscular nerves in the electron microscope, suggesting that nodal sprout growth may be modulated by the availability of endoneurial pathways sufficiently degenerated to permit reinnervation by nodal sprouts, although the initial outgrowths from nodes of Ranvier may appear in response to a growth stimulus from denervated muscle.

Animals↗

Neuromuscular transmission at terminals of sprouted mammalian motor neurones.

Motor nerve sprouting was induced in the tensor fasciae latae muscle of mice by partial denervation produced either by cutting (to prevent reinnervation) or crushing (to allow subsequent reinnervation) spinal nerve L4 unilaterally. The quantum content (m) of endplate potentials recorded intracellularly in vitro in the presence of high-Mg2+ and low-Ca2+ ion concentrations was determined up to 400 days later in non-reinnervated, reinnervated and contralateral control muscles. The muscles were then either fixed and stained with silver and cholinesterase for light microscopy, or fixed and examined in the electron microscope. The average value of m in control muscles increased by 4-5-fold as the animals matured in the 4 months following the operations. The average value of m at terminals of sprouted motor neurones in the absence of reinnervation also increased with time after partial denervation but was always less than the value in the corresponding control muscle. In electron micrographs of muscles following L4 section the nerve terminals closely apposed on average only two-thirds of the proportion of junctional folds apposed to terminals in control muscles. When muscles were reinnervated following L4 crush the average value of m at terminals of sprouted and reinnervating motor neurones equalled and sometimes exceeded m in contralateral control muscles. A proportion of muscle fibres had endplate potentials from reinnervating and sprouted axons, and the silver stain showed that these muscle fibres were innervated at the site of the original endplate. At these endplates the fraction of the total quantum content contributed by presumed sprout terminals fell significantly in the 4 months following L4 crush. It is concluded that: (i) in the absence of reinnervation, sprout terminals grow in size but a significant number never occupy all endplate site available to them; and (ii) in the presence of reinnervation axons terminals share some endplates with sprout terminals and grow at the expense of the sprout terminals which are eventually withdrawn from some shared endplates.

Action Potentials↗

The distribution of nodal sprouts in a paralysed or partly denervated mouse muscle.

The right gluteus maximus muscles of young adult mice were paralysed with botulinum toxin for up to 21 days or partly denervated by spinal root section for up to 63 days; the intramuscular and extramuscular nerves were then examined in the electron microscope in thin sections of tissue fixed conventionally or stained with zinc iodide-osmium tetroxide. Contralateral muscles were also examined as controls. The distribution of nodal sprouts in the nerves of the paralysed or partly denervated muscles was determined by calculating the mean ratios of unmyelinated to myelinated axons in nerve profiles containing different numbers of myelinated axons (intact or degenerating) and comparing these with control ratios. In paralysed muscles there was a significantly higher proportion of nerve profiles containing one or two unmyelinated axons alongside a single myelinated axon. Nerve profiles containing two or more myelinated axons did not show any increase in the proportion of unmyelinated axons. Thus, there is probably nodal sprouting in paralysed muscles which is restricted to the most distal nodes of the intramuscular nerves. In muscles partly denervated for 8 days there were significant increases in the proportion of unmyelinated axons in nerve profiles which had contained up to 5-10 myelinated axons. After 21 days of partial denervation, similar increases may have occurred in the larger intramuscular nerve profiles and after 63 days there were large increases in the proportion of unmyelinated axons in the main intramuscular nerve branches and in the extramuscular nerve. Nodal sprouting in response to partial denervation is therefore localised initially to the smaller, more distal nerve branches; at later times, some sprouts probably grow slowly in a disto-proximal direction along denervated Schwann cell pathways. The existence of nodal sprouts in paralysed muscles and their restricted distribution in paralysed and partly denervated muscles suggest that the nodal sprouting stimulus is produced by the muscle and acts only at distal nodes.

Animals↗

Comparison of effects of denervation and botulinum toxin paralysis on muscle properties in mice.

1. The properties of denervated and fully innervated but paralysed soleus and extensor digitorum longus (e.d.l.) muscles were compared in mice.2. Muscles were paralysed by single or repeated injections of botulinum toxin into the right lower leg. Contralateral muscles were denervated at the same time by sciatic nerve section. Muscles were excised 3-20 days later and direct and indirect isometric twitch and tetanic tensions measured in vitro. ACh sensitivity was assessed from isometric contractions to various concentrations of bath-applied ACh perchlorate.3. Denervated muscles as expected from the work of others developed sensitivity to ACh earlier and more rapidly than paralysed muscles. A transient peak of sensitivity was reached after 7 days of denervation in soleus and after 8 days in e.d.l. However 2 days later the sensitivity of muscles fully paralysed to nerve twitch and tetanic stimuli was not significantly different from that of denervated muscles.4. In both e.d.l. and soleus the direct tetanus/twitch ratios declined in paralysed and denervated muscles. The decline was initially more rapid in denervated than paralysed soleus, but the ratios for paralysed and denervated muscles were not significantly different after 10 days in either muscle. By this time there were no differences in the absolute strengths of the paralysed and denervated muscles.5. It is concluded that the ACh sensitivity and dynamic properties of fully paralysed mouse muscles become equal to those of denervated muscles once the acute effects associated with nerve degeneration are past. This result is consistent with the idea that normal neuronal control of extrajunctional mammalian skeletal muscle properties is mediated solely by means of activity.

Acetylcholine↗