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Biomedical subjects

E Gutmann

Publications and source records attributed to E Gutmann.

At least 19 recordsLinked to original sources

Effects of ischaemia on enzyme-activities in the soleus muscle of the rat.

1. The changes with the time of the activities of some energy-supplying enzymes and of the hydrolytic enzyme, acid phosphatase, were studied over 2 weeks of complete ischaemia, produced in the rat soleus muscle by section of the abdominal aorta and terminal devascularization, leaving nerve and tendon intact. 2. Activities of glycolytic enzymes, oxidative enzymes, hexokinase and acid phosphatase are affected in a different manner. Activities of the glycolytic enzymes, lactate dehydrogenase, triosephosphate dehydrogenase and glycerolphosphate dehydrogenase, are lowest on the 1st day and increase thereafter. The first two reach the control values again on the 4th and 14th day, respectively, while glycerolphosphate dehydrogenase reaches about 50% of the control value on the 14th day. The maximum decrease in activity of the oxidative enzymes, citrate synthase, beta-hydroxyacyl-CoA-dehydrogenase and malate dehydrogenase occurs later (4th day); thereafter their activity returns slowly to control values, but does not reach them even on the 14th day. Hexokinase activity is slightly decreased on the 1st day; then it increased and reached on the 7th day twice the control value. Thus on the 1st day the activity of the enzymes of aerobic metabolism prevail, and on the 4th day those of anaerobic carbohydrate (glucose) metabolism; the recovery of enzyme activity of aerobic oxidation occurs later. 3. Acid phosphatase activity increased from the 2nd day onwards, reaching up to 3 times the control value on the 4th day and still twice that value on the 14th day. This agrees well with the histochemical picture of acid phosphatase. 4. Histochemical changes of alkaline phosphatase activity reveal destruction of capillary endothelial cells during the first few days after operation and their later proliferation from the periphery, correlating with the loss and recovery of oxidative enzyme activity.

3-Hydroxyacyl CoA Dehydrogenases

Effect of ischemia on contractile and histochemical properties of the rat soleus muscle.

Ligature and section of the abdominal aorta results in only minor and temporary functional and metabolic changes in the slow soleus muscle of the rat. A very small decrease in maximal tetanic tension corresponds to a few scattered areas of damaged and necrotic muscle fibres, in which decreased succinic dehydrogenase and loss of phosphorylase activity was observed. A new experimental approach, i.e. ligature and section of the abdominal aorta combined with terminal devascularisation, preserving intact tendons and innervation of the muscle causes maximal muscle ischemia, followed by an almost complete loss of tetanic tension output, marked shortening of contraction time and profound morphological and histochemical changes. The decrease in succinic dehydrogenase and ATPase activities and loss of phosphorylase activity occur in the majority of degenerating muscle fibres except for a thin rim of peripheral fibres during the first 4 days. Subsequently, the contractile properties recover gradually and enzyme activities reappear in the regenerating muscle fibres simultaneously with new revascularisation. Thirty days after the operation all the parameters observed returned to control values.

Adenosine Triphosphatases

Effect of castration and testosterone administration on the neuromuscular junction in the levator ani muscle of the rat.

The ultrastructure of the neuromuscular junction (n.m.j.) of the androgen-sensitive levator ani muscle was studied in normal adult male rats, in 8-month-old rats castrated at the age of one month and in castrated rats treated with testosterone propionate (TP). Castration does not result in significant changes of the n.m.j. The density of synaptic vesicles and the postsynaptic junctional folds remain practically normal inspite of marked atrophy of the muscle. TP administration for 7 days results in marked changes in pre- and postsynaptic structures. There is slow progressive depletion of synaptic vesicles, appearance of cisternae and coated vesicles in axon terminals, and coalescence of coated vesicles with the plasma membrane. Coated vesicles are also found inside Schwann cells and among junctional folds. Dense core vesicles appear both in the axon terminals and in the postsynaptic area. Collateral sprouting of terminal axons with the formation of new immature junctions is observed. After 35 days of TP administration depletion of synaptic vesicles continues. Glycogen beta-particles, mostly freely dispersed, occasionally seen in axon terminals 7 days after TP administration, subsequently increase in number. In the endplate zone of the muscle fibre increased protein synthesis is indicated by a rapid increase in ribosomes and irregularly located myofilaments and myofibrils. The appearance of n.m.j. after testosterone administration resembles that described after nerve stimulation; the degree of change is however less pronounced.

Animals

Immunological enhancement of muscle allografts induced by serum from tolerant rats.

Serum from tolerant animals induced immunological enhancement of muscle allografts. Serum from both animals bearing tolerated skin allografts and animals in which tolerance to skin grafts disappeared was effective. No significant differences between these two types of allografts were found with respect to contractile and histochemical behaviour. Serum from animals in which skin allografts survived transiently following non-specific treatment induced weak immunological enhancement. The changes in contractile and histochemical properties of muscle allografts serve as adequate parameters for assessment of the success of immunological enhancement.

Animals

Transfer to chick embryos of incompatible DNA.

After the transfer of chicken DNA to chick embryos incompatible at the B locus a retardation of growth was observed. Retarded differentiation of muscle fibres was histochemically detected in hatched chicks by the demonstration of enzymatic activities. The manifestations of apparent weakness of the dorsal muscles resembled a "myasthenia-like" syndrome. Infection with Marek's disease virus was not responsible for the damage caused by transferred DNA.

Animals

Differentiation of myosin in soleus and extensor digitorum longus muscle in differnt animal species during development.

CA2+-ATPase activity and light chains of myosin prepared from fast and slow muscles of rat guinea-pig and rabbit were studied during development from embryonic to old age to establish further correlation with the well-known developmental changes in contraction properties of these muscles. The changes involve the slow soleus muscle much more than the fast extensor digitorum longus muscle. Myosin-ATPase activity of the soleus muscle before or at birth is higher than in the muscle of adult animals. Myosin from the soleus muscle of embryos or newborn animals reveals light chains of myosin of both fast and slow type (with a preponderance of light chains of fast type in 26-days-old rabbit embryos). During postnatal development the amount of light chains of the fast type decreases, that of the slow type increases. Myosin from the soleus muscle of adult animals contains only light chains of the slow type. However, myosin from the soleus muscle of 30-months-old rats exhibits high myosin ATPase activity and contains light chains of myosin of both slow and fast type as in perinatal development. This is in agreement with the shortening of contraction time observed in this muscle in very old age. Thus developmental differentiation of myosine in the soleus muscle is followed by a trend of levelling out of the differences between fast and slow muscles of senescent animals. No such "biphasic" development is observed with respect to the fast extensor digitorum longus muscle.

Adenosine Triphosphatases

Electrophysiological and contractile properties of the levator ani muscle after castration and testosterone administration.

Electrical and contractile properties of the levator ani muscle were studied in normal rats, in castrated rats and in castrated rats treated with testosterone. 2. No significant changes in the frequency of miniature end-plate potentials were found 6 months after castration. The frequency increased already 6 h after testosterone treatment; an increase of about 100% was observed after 7 days of testosterone treatment. 3. Castration led to a 2-fold increase of the input resistance of the muscle fibres. After 7 days of testosterone treatment the input resistance was only slightly higher than normal. 4. The weight of the muscle was decreased to 18% of the control value after 6 months castration. It increased to 46% after 7 days of testosterone treatment. 5. The muscles of castrated animals revealed a prolongation of contraction time and marked changes in maximal rate of tension development and half relaxation time. Partial recovery of these parameters was found after 7 days of testosterone treatment. 6. Long-term castration did not induce any denervation-like changes of action potential parameters, and no tetrodotoxin resistance was found in spite of marked muscle atrophy.

Anal Canal

Control of ACh sensitivity in temporarily unconnected ("decentralized") segments of diaphragm-muscle fibres of the rat.

1. After a local lesion of the diaphragm muscle, which produced a segment of innervated muscle fibres connected with an intact nerve-free segment by a region of crushed muscle fibres, the sensitivity to ACh, the presence of tetrodotoxin resistant action potentials (AP) and the transmission of AP along the muscle fibres were studied. 2. Three days after local injury of the diaphragm muscle ACh sensitivity and TTX resistance appeared in the crushed and nerve-free segments between the place of injury and the tendineous attachment. 5-7 days after injury transmission of action potentials through the damaged to the undamaged ("decentralized"), nerve-free part of the fibres is restored. High ACh sensitivity and TTX resistance of the latter segment, however, are completely lost only 20 days after the local injury. During this period contractility of the muscle remains practically unchanged. Enzymatic activities (SDH, ATPase and phosphorylase) of the damaged part were lost 3 days after crushing and recovered slowly between 7-10 days of regeneration of the diaphragm muscle fibres. 3. The experiments suggest that during regeneration of damaged muscle fibres supersensitivity to ACh remains high inspite of normal AP activity and that intracellular mechanisms may be involved in the induction and disappearance of ACh hypersensitivity.

Acetylcholine

Histochemistry of some acid hydrolases in striated muscles of the rat.

The distribution of acid phosphatase, beta-N-acetylglucosaminidase, beta-glucuronidase, and acid beta-galactosidase was studied in mm. extensor digitorum longus, soleus, and diaphragm of rats. Using the technic of semipermeable membranes activities of these enzymes were demonstrated beside cells of the interstitial tissue in muscle fibers themselves as well. Acid phosphatase displayed the highest activity which appeared in many small dots dispersed in the fiber. The activity of acid phosphatase was about 1.2 X higher in the m. soleus than in the m. extensor digitorum longus. In the latter muscle a somewhat higher activity was often found in muscle fibers displaying a higher staining for NADH tetrazolium reductase. The activity of beta-N-acetylglucosaminidase was slightly lower, that of beta-glucuronidase very weak but still discernible. The activity of acid beta-galactosidase was not ascertained in the majority of fibers. The ratio of activities measured in an area of the same size in cells of the interstitial tissue and in muscle fibers amounted in average to 2.6:1 in the case of acid phosphatase, 2.5:1 in the case of beta-N-acetylglucosaminidase, 5.7:1 in the case of beta-glucuronidase, and 44.3:1 in the case of acid beta-galactosidase. The importance of the histochemical technic in studies concerned with acid hydrolases in striated muscle fibers in normal and pathological conditions is pointed out.

Acetylglucosaminidase

Changes of acid phosphatase activity of fast and slow rat muscles during ontogenetic development.

1. The activity of acid phosphatase (AP) were studied both bio- and histochemically in fast and slow muscles of the rat during postnatal development. 2. Both biochemical and histochemical methods show at birth a similar, high AP activity in both, fast and slow muscles. From 20 days onward AP activity is higher in the slow soleus than in the fast extensor digitorum longus (EDL) muscle. 3. Fibres with high AP activity have also higher activities of oxidative enzymes and of non-specific esterase (NSE). This property is not coupled to degree of ATP-ase activity; fibres with high or low ATP-ase activity reveal similar activities with respect to AP. 4. There is a "two-phase" developmental change in the degree of enzyme activity in both the fast EDL and slow soleus muscles. AP activity is high at birth, decreases during later postnatal development and increases again in senescent muscles.

Acid Phosphatase

Different changes in contractile and histochemical properties of reinnervated and regenerated slow soleus muslces of the guinea-pig.

The changes in contraction time (CT) and histochemical muscle fibre pattern are compared with respect to ATPase activity of the slow soleus muscle of the guniea-pig during regeneration in free grafts and reinnervation after nerve interruption. Interruption of the nerve by crushing at birth results first in prolongation of CT which later returns to normal (control) values. A homogeneous fibre pattern is established related to the homogeneity of the motor units constituting the muscle. In the regenerating muscle CT and muscle fibre pattern of the graft repeat the changes during postnatal development, i.e. CT shows progressive prolongation accompanied by transformation of a heterogeneous (fibres of high and low ATPase activity) to a homogeneous (fibres of low activity only) fibre pattern. However, the regenerating muscle also shows an initial phase with slow CT. Thus the changes in reinnervation and regeneration of the muscle after birth differ, the reinnervated muscle revealing only prolongation, the regenerated muscle a temporary shortening followed by prolongation, i.e. a biphasic development of CT.

Adenosine Triphosphatases

Fate of 3H-thymidine labelled myogenic cells in regeneration of muscle isografts.

Intact and denervated extensor digitorum longus (EDL) muscles of 20-day-old inbred Lewis-Wistar rats were labelled with 3H-thymidine. Ninety minutes after the injection of the isotope 4.0% of the nuclei were labelled in the intact (i.e. innervated) and 9.6% in the muscles, denervated 3 days before administration of the isotope. The labelled EDL muscles were grafted into the bed of the previously removed EDL muscles of inbred animals and these isografts were studied 30 days later. In the EDL muscles, regenerated from innervated isografts only occasionally labelled endothelial cells were found whereas in the muscles regenerated from denervated isografts also parenchymal muscle nuclei were regularly labelled. The incidence of labelled nuclei in the regenerated EDL muscles was, however, about 20 times lower than in the donor EDL muscles. The presen experiments provide a direct proof of utilization of donor satelite cell nuclei for regeneration in grafted muscle tissue. With respect to the low incidence of labelled nuclei in regenerated EDL muscles, other sources of cells apparently also contribute to the regeneration process.

Animals