Topic:Adenosine Triphosphate (ATP)
Cardiovascular adaptations to exercise and training. The cardiovascular system provides the link between pulmonary ventilation and oxygen usage at the cellular level. During exercise, efficient delivery of oxygen to working skeletal and cardiac muscles is vital for maintenance of ATP production by aerobic mechanisms. The equine cardiovascular response to increased demand for oxygen delivery during exercise contributes largely to the over 35-fold increases in oxygen uptake that occur during submaximal exercise. Cardiac output during exercise increases greatly owing to the relatively high heart rates that are achieved during exercise. Heart rate i...
Metabolic response of equine muscle to intermittent maximal exercise. Four thoroughbred horses performed 4 gallops (G1-G4) with intervals of 5 min. With one exception, gallops were sustained at maximal speed over 620 m. Muscle biopsy samples of the middle gluteal and brachiocephalicus were taken before, during, and after exercise and assayed for ATP and intermediary metabolites. The results showed a major involvement of the brachiocephalicus, in addition to the middle gluteal, during galloping. In three horses, who were clearly fatigued, muscle ATP decreased by up to 50% by the end of G4. This was matched by an equal rise in inosine 5'-monophosphate. Pronounced ...
Comparison of fiber types in skeletal muscles from ten animal species based on sensitivity of the myofibrillar actomyosin ATPase to acid or copper. Comparisons were made of the histochemical characteristics of skeletal muscle from 10 animal species. The basic comparison was made from the staining patterns for the myofibrillar actomyosin ATPase produced by preincubation of fresh frozen cross-sections of muscle at alkaline pH (10.30) or acid pH (4.60) with those produced by preincubation in media containing Cu2+ at alkaline pH (10.30), near neutral pH (7.40), or acid pH (4.60). Muscle sections were also stained for reduced nicotinamide adenine dinucleotide tetrazolium reductase and alpha-glycerophosphate dehydrogenase to provide an indicati...
Microheterogeneity of type II cAMP-dependent protein kinase in various mammalian species and tissues. Excluding autophosphorylated species, at least six forms of the regulatory subunit of type II cAMP-dependent protein kinase (RII) from various mammalian tissues were identified by sodium dodecyl sulfate (SDS) gel electrophoresis of purified samples and of crude preparations photoaffinity labeled with 8-azido[32P] cAMP and by gel filtration. After autophosphorylation some heart RII forms termed type IIA (bovine, porcine, equine, and dog) shifted to a more slowly migrating band on SDS gels while others termed type IIB (rat, guinea pig, rabbit, and monkey) did not detectably shift. Both subclasse...
Biochemical and functional characterization of lymphocytes from a horse with lymphosarcoma and IgM deficiency. Neoplastic lymphocytes from a horse with lymphosarcoma and IgM deficiency were analyzed for ability to grow in culture; surface and cytoplasmic IgM; functional activity in blastogenesis, cytoxicity, and suppressor assays; and activities of six enzymes involved in purine and pyrimidine metabolism. The cells lacked surface and cytoplasmic IgM. They had elevated activity of adenosine deaminase and reduced activity of purine nucleoside phosphorylase. Neoplastic cells were nonresponsive in blastogenesis assay and did not kill allogeneic lymphocyte target cells or YAC-1 targets in a lectin-dependent...
Platelet function, size and yield in whole blood and in platelet-rich plasma prepared using differing centrifugation force and time in domestic and food-producing animals. The effects of centrifugation force and time upon platelets function, mean platelet volume and platelet yield were compared with whole blood platelet counts and size in citrated blood samples from the bovine, canine, caprine, equine, feline, ovine and porcine species. The results were similar, for a given species, irregardless of sample volume. Bovine, caprine, feline and ovine platelet yields and mean platelet volumes were maximal when platelet-rich plasma was prepared using longer centrifugation times and lower gravitational forces. Canine, equine and porcine platelet yields and mean platele...
Increased susceptibility of fibroblasts from horses with severe combined immunodeficiency to growth inhibition by 2′-deoxyadenosine. The effect of adenosine, deoxyadenosine, guanosine, and deoxyguanosine on the growth rate of fibroblasts derived from normal horses, horses heterozygous for the severe combined immunodeficiency (SCID) trait (heterozygotes), and horses with SCID was studied. All four purines were found to inhibit growth in a dose-dependent manner, but only adenosine and deoxyadenosine were inhibitory at concentrations of less than 100 microM. No statistical difference in sensitivity to adenosine was detected between normal and SCID fibroblasts. Fibroblasts from SCID horses were, however, more sensitive to the g...
Sodium and potassium ion-dependent change in oligomerization of Na,K-ATPase in C12E8 detected by low-angle laser light scattering technique in combination with high performance porous silica-gel chromatography. Approximate molecular weights and the subunit structures of Na,K-ATPase from horse kidney were estimated by means of the combination of porous silica gel chromatography, laser light scattering (LS) and refractive index (RI) measurements in C12E8. When the enzymes were eluted with NaCl- or KCl-containing solution, 3 or 4 protein peaks, respectively were detected except that of low molecular weight range. These peaks were tentatively named Na-1, Na-2, Na-2', Na-3 (NaCl-containing eluents), K-1, K-2, K-3 (KCl-containing eluents), respectively. Na,K-ATPase and K-p-nitrophenylphosphatase activities...
Concentration of nucleotides and deoxynucleotides in peripheral and phytohemagglutinin-stimulated mammalian lymphocytes. Effects of adenosine and deoxyadenosine. Concentrations of purine and pyrimidine ribonucleotides were measured with HPLC in lymphocytes of man, horse, pig and sheep and in rat thymocytes. The ATP concentration was highest in lymphocytes of all species and about 850 pmol/10(6) cells in human and equine lymphocytes, higher in porcine and lower in ovine lymphocytes and rat thymocytes. The GTP concentration was comparable in human, equine and porcine lymphocytes, but lower in ovine lymphocytes. ATP concentration was also measured in lymphocytes of man, horse and pig with a luciferin-luciferase assay. During culturing with or without phyt...
Regional myocardial blood flow and coronary vascular reserve in unanesthetized ponies during pacing-induced ventricular tachycardia. To examine the effects of tachycardia on coronary circulation, transmural distribution of myocardial blood flow (MBF, 15-micron diameter radionuclide-labeled microspheres) was studied in six healthy adult ponies at rest (heart rate = 60 +/- 7 beats . min-1), during ventricular pacing at 150 and 200 beats . min-1, as well as with ventricular pacing at 250 beats . min-1 before and during maximal coronary vasodilatation (iv adenosine infusion; 4 mumole . kg-1 . min-1). Mean aortic pressure and cardiac output were unchanged from control values with ventricular pacing. Whereas ventricular pacing at...
A comparative study of the effect of triazine herbicides on alcohol dehydrogenases isolated from various sources. The studied herbicides (terbutylazine, simazine) inhibit the activity of plant, animal, and yeast alcohol dehydrogenases. The inhibition constant Ki for alcohol dehydrogenase (ADH) isolated from peas and bakers' yeast equals approximately 10(-4) M, and that for ADH isolated from horse liver is of the order of 10(-5) M. The character of inhibition for all the herbicides studied for the reaction catalyzed by pea, liver, and yeast ADH is always noncompetitive toward ethanol and competitive with respect to NAD. The inhibition constants for the enzyme isolated from peas are pH independent. The inte...
Cytotaxin-induced cAMP peak in granulocytes: its relationship to crawling movements, chemokinesis and chemotaxis. The relationship between the short transient intracellular increase in cAMP levels on the one hand and chemotaxis or crawling movements on the other hand was investigated using human and equine granulocytes. C5ades arg, f-met-leu-phe, human serum albumin and immunoglobulin were used as stimulating agents. There was no strict correlation between the induction of crawling movements or of chemokinesis in general and the generation of the cAMP peak. But there was so far a strict parallelism between the occurrence of the chemotactic response and the cAMP peak. However, the magnitude of the peak was...
Energy metabolism in the erythrocytes of thoroughbred horses connected with perinatal physiological hemolysis. 1. The metabolism in the erythrocytes of thoroughbred horses in a sequential study from umbilical cord to the 1st month was investigated. 2. Emphasis was put on hemolytic period at which: (a). PFK, GSH-Px and GSH play a significant role. (b). There is a lower glucose consumption determined by a decreased activity in several enzymatic steps. (c). Singularly high concentrations of 2-3DPG and ATP were detected. 3. It has been suggested that the metabolic adjustments were achieved by an increased activity of the hexose monophosphate shunt, G-3PD and AK.
The optimum pH of renal adenosine triphosphatase in rats: influence of vanadate, noradrenaline and potassium. In the presence of vanadate, the optimum pH of renal (Na+, K+)-ATPase in rats is reduced and lies in the range of intracellular pH. This explains the difference in optimum pH observed with ATP extracted from equine muscle. Removal of vanadate from such ATP (with noradrenaline) raises the optimum to the accepted range obtained with synthetic ATP. Changes in the sensitivity of the enzyme to potassium concentration contribute to the alterations in optimum pH. The optimum pH of Mg-ATPase is unaffected by vanadate. Since vanadate may be an intracellular regulator of (Na+, K+)-ATPase changes of opti...
Effects of adenosine and deoxyadenosine on PHA-stimulation of lymphocytes of man, horse and pig. 1. Adenosine inhibits thymidine and uridine incorporation of PHA-stimulated lymphocytes of man and horse at concentrations higher than 50 and 10 microM, respectively. Deoxyadenosine is inhibitory at concentrations higher than 100 microM. Thymidine and uridine incorporation of porcine lymphocytes are elevated 5-7-fold by 25-100 microM adenosine, deoxyadenosine, inosine and hypoxanthine. Leucine incorporation of PHA-stimulated lymphocytes was affected by adenosine and deoxyadenosine in the same way, but to a lower extent. 2. Effects of adenosine and deoxyadenosine were more pronounced at shorter...
The interaction of equine platelet tropomyosin with skeletal muscle actin. Whereas skeletal muscle tropomyosin binds strongly to muscle F-actin in a buffer containing 30 mM KCl and 1-2 mM free Mg2+, equine platelet tropomyosin only binds stoichiometrically (1 tropomyosin molecule per 6 actin monomers) at higher Mg2+ concentrations (7-8 mM free Mg2+). At low free Mg2+ concentrations (1.5 mM) the binding of the platelet protein is only marginally increased by raising the KCl concentration to an optimal value (0.10-0.20 M). This weaker binding can be attributed to the relatively poor head-to-tail polymerization of platelet tropomyosin and its fewer actin-binding sites. ...
Activation of pigeon erythrocyte adenylate cyclase by cholera toxin. Partial purification of an essential macromolecular factor from horse erythrocyte cytosol. A cytosolic, macromolecular factor required for the cholera toxin-dependent activation of pigeon erythrocyte adenylate cyclase and cholera toxin-dependent ADP-ribosylation of a membrane-bound 43,000 dalton polypeptide has been purified 1100-fold from horse erythrocyte cytosol using organic solvent precipitation and heat treatment. This factor, 13,000 daltons, does not absorb to anionic or cationic exchange resins, is sensitive to trypsin or 10% trichloroacetic acid and is not extractable by diethyl ether. Activation of adenylate cyclase by cholera toxin requires the simultaneous presence of AT...
Uptake of nucleotides and catecholamines by chromaffin granules from pig and horse adrenal medulla. The uptake of nucleotides and catecholamines into chromaffin granules from adrenals of pigs and horses is similar to that previously seen in bovine chromaffin granules. The rate of [3H]ATP uptake at 2 mM-ATP concentration was 0.42 +/- 0.06 and 0.15 +/- 0.02 nmol/mg protein/min for pig and horse granules, respectively. The apparent Km's were 1.37 mM for pig granules, 0.89 mM for horse granules, and 1.2 mM for ox granules. The sensitivity of the uptake for nucleotides and catecholamine to specific inhibitors was found to be similar in granules from pig and ox, indicating that the same mechanisms...
In vitro of adenosine on lymphocytes and erythrocytes from horses with combined immunodeficiency. The effect of adenosine on the mitogenic response of peripheral blood lymphocytes (PBL) and on the nucleotide pools of erythrocytes from normal horses, horses heterozygous for the combined immunodeficiency (CID) trait (carriers), and foals with CID was studied. When PBL from normal, carrier, and CID horses were stimulated by phytohemagglutinin (PHA), concanavalin A, or pokeweed mitogen, [3H]thymidine uptake was inhibited by adenosine (0.1 microM) to 1.0 mM) in a dose-dependent manner. Adenosine (100 microM) mediated inhibition of [3H]thymidine uptake was prevented in both normal and carrier ho...
A comparative histochemical study of intrinsic laryngeal muscles of ungulates and carnivores. The intrinsic laryngeal muscles of the horse, donkey, sheep, ox, pig, dog and cat were examined for myosin ATPase, following acid and alkali pre-incubation, SDH and M-alphaGPDH activities. In all laryngeal muscles two fibre types, betaR and alphaR, belonging to slow and fast-contracting, fatigue-resistant motor units (types S and FR) were present in different proportions. The alphaW fibre type, belonging to fast-contracting and fatigue-resistant motor units was absent (type FF). The alphaR fibres of the dog and the cat were subdivided into groups by the various degrees of acid stable myosin AT...
Differences in the histochemical properties of skeletal muscles of different breeds of horses and dogs. Histochemical profiles of individual muscle fibres were established using myosin adenosine triphosphatase (myosin ATPase), succinate dehydrogenase (SDHase), and glycogen phosphorylase (GPase) reactions in three muscles (semitendinosus, diaphragm, and pectoralis transversus) of the horse and dog. The major histochemical difference between fibres lies in their myosin ATPase activity; fibres can be subdivided into those with a high and those with a low activity. In horse muscle, all fibres have a high activity of GPase. In the diaphragm and pectoralis transversus, all fibres have a high SDHase ac...
The presence of two (Na+ + K+)-ATPase inhibitors in equine muscle ATP: vanadate nad a dithioerythritol-dependent inhibitor. A potent inhibitor of (Na+ + K+)-ATPase activity was purified from Sigma equine muscle ATP by cation- and anion-exchange chromatography. The isolated inhibitor was identified by atomic absorption spectroscopy and proton resonance spectroscopy to be an inorganic vanadate. The isolated vanadate and a solution of V2O5 inhibit sarcolemma (Na+ + K+)-ATPase with an I50 of 1 micrometer in the presence of 1 mM ethyleneglycol-bis-(beta-aminoethylether)-N,N'-tetraacetic acid (EGTA), 145 mM NaCl, 6mM MgCl2, 15 mM KCl and 2 mM synthetic ATP. The potency of the isolated vanadate is increased by free Mg2+. ...
Characterization of protein phosphokinase activities in horse thyroid nuclei. The distribution of protein phosphokinase (EC 2.7.1.37) activities has been established in horse thyroid nuclei. The presence of several enzyme activities has been demonstrated, two of which are clearly distinct. The first one acts on histone as substrate and is activated by cyclic AMP. Physico-chemical properties of this nuclear cyclic AMP-dependent histone kinase and of the cytosol histone kinase are different, demonstrating the absence of a contamination from the cytosol. The second enzyme acts on casein as substrate and is not stimulated by cyclic AMP POR CYCLIC GMP. The findings are consi...
Evaluation of adenosine deaminase and other purine salvage pathway enzymes in horses with combined immunodeficiency. Foals with combined immunodeficiency had normal levels of purine salvage pathway enzymes, including adenosine deaminase, nucleoside phosphorylase, and xanthine oxidase.