Porcine skeletal muscle for physiological studies

Summary The common digital extensor (lateral head) is one of the few porcine muscles of convenient dimensions for in vitro studies of intact cells. Its removal and use in physiological studies are described.Acknowledgments. The pigs used in this investigation were provided by Drs D. G. Topel, L. L. Christian and G. A. Gronert whose cooperation is appreciated. Gratitude is also expressed to Dr N. G. Ghoshal for assistance with anatomical identifications.     

Porcine skeletal muscle for physiological studies.

The common digital extensor (lateral head) is one of the few porcine muscles of convenient dimensions for in vitro studies of intact cells. Its removal and use in physiological studies are described.     

Control of respiration in skeletal muscle at rest

The suggestion is made that, under resting conditions in situ, muscle cell respiration is dependent on the way O2 and substrates are distributed to the cells by the microcirculation. (Delivery is measured as arterial-blood concentration multiplied by flow to the organ.) Microscale heterogeneity of this distribution, which is more marked but less stable than the more easily demonstrated larger-scale heterogeneity (0.1 to 0.5-g sampling grain), might indeed ration O2 and substrates in a large population of the cells of a resting organ at any given moment, and microscale heterogeneity of distribution may thus take part in the normal control of cell respiration.     

Demonstration of retinoic acid isomers in human urine under physiological conditions

Untransformed retinoic acid has never been demonstrated in human excreta under normal physiological conditions. We have developed a two-step liquid chromatographic system for the demonstration of subnanogram amounts of this compound in human urine without administration of any precursor.     

Demonstration of retinoic acid isomers in human urine under physiological conditions

Summary Untransformed retinoic acid has never been demonstrated in human excreta under normal physiological conditions. We have developed a two-step liquid chromatographic system for the demonstration of subnanogram amounts of this compound in human urine without administration of any precursor.The work was supported by a grant of the N.F.S.R. to W.L.     

Control of adenine nucleotide metabolism and glycolysis in vertebrate skeletal muscle during exercise

The turnover of adenosine triphosphate (ATP) in vertebrate skeletal muscle can increase more than a hundredfold during high-intensity exercise while the content of ATP in muscle may remain virtually unchanged. This requires that the rates of ATP hydrolysis and ATP synthesis are exactly balanced despite large fluctuations in reaction rates. ATP is regenerated initially at the expense of phosphocreatine (PCr) and then mainly through glycolysis from muscle glycogen. The increased ATP turnover in contracting muscle will cause an increase in the contents of adenosine diphosphate (ADP), adenosine monophosphate (AMP) and inorganic phosphate (P_i), metabolites that are substrates and activators of regulatory enzymes such as glycogen phosphorylase and phosphofructokinase. An intracellular metabolic feedback mechanism is thus activated by muscle contraction. How muscle metabolism is integrated in the intact body under physiological conditions is not fully understood. Common frogs are suitable experimental animals for the study of this problem because they can readily be induced to change from rest to high-intensity exercise, in the form of swimming. The changes in metabolites and effectors in gastrocnemius muscle were followed during exercise, post-exercise recovery and repeated exercise. The results suggest that glycolytic flux in muscle is modulated by signals from outside the muscle and that fructose 2,6-bisphosphate is a key signal in this process.     

Histochemical demonstration of 1,4-amylophosphorylase in the human testis under normal and pathological conditions

Riassunto La 1–4 amilofosforilasi è presente nel testicolo umano normale ove è localizzata nell'epitelio tubulare e in alcuni elementi peritubulari. Nella aplasia germinale la reazione è intensamente positiva solo in sede peritubulare, nell'arresto maturativo solo nel tubulo. L'enzima è probabilmente in rapporto con l'energia necessaria per la spermatogenesi e con la funzione contrattile delle cellule peritubulari.

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Supported by grant No. 1247 of Consiglio Nazionale delle Ricerche Roma, Italia.     

The autooxidation of 2,3,5,6-tetrahydroxy-2,5-cyclohexadiene-1,4-dione under physiological conditions

Summary A radical anion of 2,3,5,6-tetrahydroxy-2,5-cyclohexadiene-1,4-dione was detected using the EPR technique by complexing with Zn ions. Hydrogen peroxide, superoxide radical anion and hydroxyl radical were also detected in the reaction mixture. Kinetic study and product distribution indicated a probable mixed type one- and two-electron transfer mechanism. A possible relationship between the autooxidation process and the biological activity of substituted quinones was suggested.     

Studies on muscle fibre splitting in skeletal muscle

Summary Autoradiographic, stereological and histological studies have been carried out to determine the origin of muscle fibre splitting which supposedly occurs during muscle hypertrophy. The results obtained clearly indicate that the supposedly split fibres are a transient response probably derived from satellite cells and are not derived from pre-existing fibres by true splitting. Similarly, increases in muscle fibre size are not achieved by recruitment of satellite structures as indicated by lack of myonuclear recruitment.Acknowledgment. This work was carried out with the aid of a grant from the Medical Research Council of Great Britain. The authors are grateful for the excellent technical assistance of Miss H. Caulton, M.J. Wild and M. Fenner.     

A DNA extraction procedure practicable under field work conditions

Summary A DNA extraction procedure is described which is designed to be used in field work. A crude initial cell precipitate, which is easily obtained, may endure storage of months.Acknowledgment. The skilful technical assistance by MissB. Kunz is gratefully acknowledged.     

Pleiotropic effects of sphingolipids in skeletal muscle

Studies of the last two decades have demonstrated that sphingolipids are important signalling molecules exerting key roles in the control of fundamental biological processes including proliferation, differentiation, motility and survival. Here we review the role of bioactive sphingolipids such as ceramide, sphingosine, sphingosine 1-phosphate, ganglioside GM3, in the regulation of skeletal muscle biology. The emerging picture is in favour of a complex role of these molecules, which appear implicated in the activation of muscle resident stem cells, their proliferation and differentiation, finalized at skeletal muscle regeneration. Moreover, they are involved in the regulation of contractile properties, tissue responsiveness to insulin and muscle fiber trophism. Hopefully, this article will provide a framework for future investigation into the field, aimed at establishing whether altered sphingolipid metabolism is implicated in the onset of skeletal muscle diseases and identifying new pharmacological targets for the therapy of multiple illnesses, including muscular dystrophies and diabetes. Received 30 April 2008; received after revision 19 June 2008; accepted 14 July 2008     

Evidence for a novel racemization process of an asparaginyl residue in mouse lysozyme under physiological conditions

We examined chemical reactions in mouse lysozyme after incubation under physiological conditions (pH 7 and 37°C). After incubation for 8 weeks, racemization was observed specifically at Asn127 among the 19 Asp/Asn residues in mouse lysozyme. Furthermore, analysis of the primary structure showed that the racemized residue was not Asp, but Asn, which demonstrates that deamidation and isomerization did not occur. These results mean that this racemization occurs without forming a succinimide intermediate. This is the first example of D-asparaginyl formation in a protein occurring during the racemization process under physiological conditions.Received 16 September 2004; received after revision 26 October 2004; accepted 12 November 2004     

Glutaminase activity in rat skeletal muscle mitochondria

Isolated rat skeletal muscle mitochondria took up about 40-ng-atoms O per mg protein, with glutamine as the only respiratory substrate. The mitochondria incubated in the presence of glutamine and KCN formed both ammonia and glutamate in equivalent amounts. The experiments reported here provide suggestive evidence that rat skeletal muscle mitochondria contain glutaminase (L-glutamine amidohydrolase EC 3.5.1.2.) activity.