Na+, K(+)-ATPase activity and ATP concentration in horses of the Wielkopolski breed in relation to age

This study aimed at determining relationships between the age of the Wielkopolski horses, ATP in whole blood and in the erythrocytes, and between erythrocyte Na+, K(+)-ATPase activity, and serum concentrations of mineral components. ATP was measured in whole blood and in erythrocytes by HPLC method. Serum concentrations of Ca2+ and Mg2+ were measured spectrophotometrically, while Na+ and K+ by flame photometry. In horses aged from 4 to 48 months, a dynamic decrease in ATP activity was found. Erythrocyte Na+, K(+)-ATPase activity decreased proportionally with the decrease in ATP activity. The results of this study may enable planning physical effort of horses at optimum use of energetic efficiency of their erythrocytes and mineral components in relation to their age.     

Effects of oral potassium supplementation on acid-base status and plasma ion concentrations of horses during endurance exercise

OBJECTIVE: To compare effects of oral supplementation with an experimental potassium-free sodium-abundant electrolyte mixture (EM-K) with that of oral supplementation with commercial potassium-rich mixtures (EM+K) on acid-base status and plasma ion concentrations in horses during an 80-km endurance ride. ANIMALS: 46 healthy horses. PROCEDURE: Blood samples were collected before the ride; at 21-, 37-, 56-, and 80-km inspection points; and during recovery (ie, 30-minute period after the ride). Consumed electrolytes were recorded. Blood was analyzed for pH, PvCO2, and Hct, and plasma was analyzed for Na+, K+, Cl-, Ca2+, Mg2+, lactate, albumin, phosphate, and total protein concentrations. Plasma concentrations of H+ and HCO3-, the strong ion difference (SID), and osmolarity were calculated. RESULTS: 34 (17 EM-K and 17 EM+K treated) horses finished the ride. Potassium intake was 33 g less and Na+ intake was 36 g greater for EM-K-treated horses, compared with EM+K-treated horses. With increasing distance, plasma osmolarity; H+, Na+, K+, Mg2+, phosphate, lactate, total protein, and albumin concentrations; and PvCO2 and Hct were increased in all horses. Plasma HCO3-, Ca2+, and Cl- concentrations were decreased. Plasma H+ concentration was significantly lower in EM-K-treated horses, compared with EM+K-treated horses. Plasma K+ concentrations at the 80-km inspection point and during recovery were significantly less in EM-K-treated horses, compared with EM+K-treated horses. CONCLUSIONS AND CLINICAL RELEVANCE: Increases in plasma H+ and K+ concentrations in this endurance ride were moderate and unlikely to contribute to signs of muscle fatigue and hyperexcitability in horses.     

盐胁迫对芨芨草种子萌发苗Na+,K+含量与质膜H+-ATPase活性的影响

以芨芨草(Achnatherumsplendens)种子萌发苗为试验材料,分别以不同浓度NaCl和Na₂SO₄进行胁迫,通过对芨芨草叶片和根系中Na⁺,K⁺含量以及质膜H⁺-ATPase活性进行测定,以探讨盐胁迫对芨芨草中Na⁺,K⁺分布以及质膜H⁺-ATPase活性的影响。结果表明:随着NaCl和Na₂SO₄浓度的增加,芨芨草根系和叶片的Na⁺含量增加,K⁺含量下降,K⁺/Na⁺比值下降,根系中质膜H⁺-ATPase活性增加;在NaCl和Na₂SO₄胁迫下,芨芨草叶片中Na⁺含量显著低于根系,K⁺含量显著高于根系,叶片的K⁺/Na⁺比值均大于1并明显高于根系,根系的质膜H⁺-ATPase活性显著高于叶片;与NaCl相比,Na₂SO₄胁迫下,芨芨草根系向叶片的离子选择性运输系数(TS_K,Na)较高,叶片和根系的质膜H⁺-ATPase活性明显高于相同浓度的NaCl胁迫组。与NaCl胁迫相比,芨芨草对Na₂SO₄胁迫的适应性更强。     

磷对奶牛红细胞膜Na^＋,K^＋—ATPase活性影响的观察

观察了土壤、牧草低磷情况下，可影响奶牛血清磷含量。血清磷含量可影响红细胞膜结构与功能。血清磷含量减少会导致Ｎａ＾＋，Ｋ＾＋－ＡＴＰａｓｅ活性降低，进而影响Ｃａ＾２＋－ＡＴＰａｓｅ活性和Ｍｇ＾２＋－ＡＴＰａｓｅ活性。     

Effects of training on potassium homeostasis during exercise and skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged Dutch Warmblood horses

OBJECTIVE: To investigate the effects of moderate short-term training on K+ regulation in plasma and erythrocytes during exercise and on skeletal muscle Na+,K(+)-ATPase concentration in young adult and middle-aged horses. ANIMALS: Four 4- to 6-year-old and four 10- to 16-year-old Dutch Warmblood horses. PROCEDURE: The horses underwent a 6-minute exercise trial before and after 12 days of training. Skeletal muscle Na+,K(+)-ATPase concentration was analyzed in gluteus medius and semitendinosus muscle specimens before and after the 12-day training period. Blood samples were collected before and immediately after the trials and at 3, 5, 7, and 10 minutes after cessation of exercise for assessment of several hematologic variables and analysis of plasma and whole-blood K+ concentrations. RESULTS: After training, Na+,K(+)-ATPase concentration in the gluteus medius, but not semitendinosus, muscle of middle-aged horses increased (32%), compared with pretraining values; this did not affect the degree of hyperkalemia that developed during exercise. The development of hyperkalemia during exercise in young adult horses was blunted (albeit not significantly) without any change in the concentration of Na+,K(+)-ATPase in either of the muscles. After training, the erythrocyte K+ concentration increased (7% to 10%) significantly in both groups of horses but did not change during the exercise trials. CONCLUSIONS AND CLINICAL RELEVANCE: In horses, the activation of skeletal muscle Na+,K(+)-ATPase during exercise is likely to decrease with age. Training appears to result in an increase in Na+,K(+)-ATPase activity in skeletal muscle with subsequent upregulation of Na+,K(+)-ATPase concentration if the existing Na+,K(+)-ATPase capacity cannot meet requirements.     

Analysis of muscle elements, water, and total lipids from healthy dogs and Labrador retrievers with hereditary muscular dystrophy

Skeletal muscles from healthy dogs and Labrador Retrievers with hereditary muscular dystrophy were examined morphologically and histochemically and were analyzed biochemically for Na+, K+, Ca2+, Mg2+, Zn2+, Cu2+, Cl-, total muscle water, and total neutral lipid content. Flame atomic absorption spectrophotometer was used for elemental quantitation of hydrochloric acid tissue extracts. Muscle samples from dystrophic dogs contained substantially increased concentrations of Na+, Ca2+, Zn2+, Cu2+, and Cl-, and a considerable reduction in the content of K+ and Mg2+ compared with samples from healthy dogs. Total muscle water and total fat content was higher in muscles from dystrophic dogs. Most muscle samples from dystrophic dogs had a type-2 fiber deficiency and an increase in number of fibers with internalized nuclei.     

铜缺乏对奶牛红细胞及组织细胞膜ATP酶活性的影响

为探讨铜缺乏对奶牛红细胞及组织细胞膜ATP活性的影响，在调查铜缺乏奶牛水土食物链微量元素状态的基础上，首次测度了病区6头发病犊奶牛（Ⅱ组）及该地区20头同年生临床健康奶牛（IH组，血清铜低于0.56mg/kg）红细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶及Mg^2 -ATP酶的活性，对20头IH组奶牛进行为期80d的补铜试验（含铜40mg/kg干饲料），以20头健区同年生健康奶犊牛作为对照（HH组，全血铜大于０.85mg/L）。同时测试了剖杀后６头病牛及健康牛组织细胞膜ＡＴＰ酶的活性。结果表明：Ⅱ组奶牛红细胞膜ＡＴＰ酶活性显著低于ＨＨ组及ＩＨ组。补铜后４０d，ＩＨ组奶牛红细胞膜Na^ -K^ -ATP酶均显著升高，病牛肝、脾、肾、大脑、小脑、淋巴结等组织细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶活性显著低于对照组。结论：铜缺乏较为严重地影响了奶牛红细胞及组织膜ＡＴＰ酶的活性，抑制了这些组织的正常生理功能。     

Right-side torsion of the abomasum in dairy cows: classification of severity and evaluation of outcome

A retrospective analysis was made of 100 cases of surgically corrected right-side torsion of the abomasum (RTA) in dairy cows. A classification for degrees of severity of RTA was developed, based on the amount of fluid sequestered in the abomasum. Serum concentrations of Na+, K+, AND Cl- decreased with increasing severity of RTA. Cows in the most severely affected classification were markedly hypochloremic and hypokalemic. The PCV, plasma protein concentration, and pulse rate increased with increasing severity of RTA. Statistically significant correlations were observed between postsurgical outcome and classifications of RTA, serum Cl- concentration, and pulse rate. The outcome was not statistically related to serum concentrations of Na+ or K+, PCV, or plasma protein concentration.     

Preliminary Studies on the Concentration of Na+,K+-ATPase in Skeletal Muscle of Draught Cattle in Mozambique: Effect of Sex,Age and Training

The effect of training on the potential for work in draught cattle was assessed by measuring the Na⁺,K⁺-ATPase in the muscle cell membrane and the elevation in the concentration of K⁺ in plasma during exercise. Biopsies of the semitendinosus muscle and venous blood samples were taken from the cattle used for draught work in Mozambique. No differences were found in the plasma ion or Na⁺,K⁺-ATPase concentrations in samples taken from Nguni, Africander and Angoni breeds. There were no significant differences in plasma ions (Na⁺, K⁺ and Cl^–) or muscle Na⁺,K⁺-ATPase concentrations between the Angoni males and females, although the males showed an increase in Na⁺,K⁺-ATPase with age, while the females showed a decrease. The increase in males might be attributed to their higher level of activity in the herds than that of females. After a training period of 15 days, a significant increase in Na⁺,K⁺-ATPase concentration in semitendinosus muscle was found in Angoni cattle. In females, this was significant after 8 days of training (about 30%); in males after 15 days of training (about 16%). On day 15, there was a reduction in the elevation of plasma K⁺ during the 2 h of draught work, indicating an increased capacity of the Na⁺,K⁺ pumps to maintain the extracellular K⁺ concentration in working muscles and a possible delay in the moment of fatigue.     

回—直肠吻合荷术猪水和盐代谢规律研究

本试验用回—直肠吻合猪进行手术前后的对比试验 ,旨在研究回—直肠吻合术对于猪的水和盐代谢规律所产生的影响。以试验日粮分别于回—直肠吻合手术5天前和手术后15天定时、定量饲喂大莱公猪 ,连续收集3天试验猪的粪、尿样品 ,分别测定粪中的粗蛋白质、粗纤维及尿中的粗蛋白质和钾、钠、氯、钙离子含量。结果表明 :回—直肠吻合手术后 ,试验猪所排粪和尿的总量以及粪中的粗蛋白质、水分 ,尿中的粗蛋白质、钙离子含量与术前相比变化极显著(P<0.01) ;手术前后粪中的粗纤维及尿中的钠离子、氯离子含量差异显著(P<0.05) ;而手术对于尿中钾离子含量的影响不大(P>0.05)。     

Effects of Pasteurella haemolytica leukotoxin on cultured bovine lymphoma cells

Leukotoxin activity from culture supernatants of Pasteurella haemolytica serotype 1 in logarithmic growth phase caused rapid (less than 5 min) release of intracellular K+, uptake of extracellular Ca2+, and swelling of cultured bovine lymphoma cells (BL3 cells). Release of 51CrO4(2-) and lactate dehydrogenase (LDH) from BL3 cells began after 15 minutes of incubation with leukotoxin at 37 C and was completed between 60 and 120 minutes of incubation. In addition, leukotoxin exposure of BL3 cells resulted in cell aggregation and adherence to glass surfaces. Scanning electron microscopy indicated that after 10 minutes of leukotoxin exposure, BL3 cells increased in size, and large membrane defects developed between 20 and 60 minutes of exposure. The rate of release of LDH from leukotoxin-exposed BL3 cells was proportional to the amount of leukotoxin added. At high cell concentrations, the activity of LDH released at completion was directly proportional to the amount of leukotoxin added. Leukotoxin-induced release of LDH required a divalent cation, whereas K+ release and cell swelling did not. The addition of Ca2+, Mn2+, and Ba2+ resulted in increased leukotoxin-induced release of LDH. Divalent cation concentrations of 0.5 to 2.5 mM resulted in 50% of maximal stimulation. Ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid blocked increased release of LDH caused by Ca2+ addition, but had no effect on K+ release or cell swelling. Leukotoxin action on BL3 cells (K+ release, cell swelling, Ca2+ uptake, and release of LDH) was prevented by incubation at 4 C.     

Insulin accelerates recovery from QRS complex widening in a frog heart model of hyperkalemia

Hyperkalemia is one of the most common electrolyte disorders. By injecting various concentrations of potassium chloride (KCl) solutions intravenously into bullfrogs, we demonstrated characteristic electrocardiogram (ECG) abnormalities of hyperkalemia in frog hearts. The widened QRS complexes induced by 100 mM KCl injection were accompanied by an increase in the resting membrane potential in cardiomyocytes and a decreased slope of phase 0 in the action potential. Recording both ECG waveforms and the cardiac action potential enabled us to reveal the mechanisms of hyperkalemia-induced ECG abnormalities. Additionally, pre-treatment with insulin, a powerful stimulator of Na⁺/K⁺-ATPase activity, significantly accelerated the recovery from the widened QRS complexes in the ECG, demonstrating a pronounced shift of extracellular potassium ions into the intracellular space.     

小型猪特异性麻醉颉颃剂对大鼠不同脑区Na+,K+-ATP酶活性的影响

为探讨Na+,K+-ATP酶在小型猪特异性麻醉颉颃剂催醒过程中的作用,144只Wistar大鼠随机分为对照组和试验组,对照组和试验组再分为早期催醒组、中期催醒组和晚期催醒组.用分光光度法测定不同脑区Na+,K+-ATP酶活性.结果显示,大鼠在不同时期注射小型猪特异性麻醉颉颃剂后,大鼠不同脑区的Na+,K+-ATP酶活性均被激活,且Na+,K+-ATP酶活性的这种变化趋势与大鼠行为学的变化相一致.表明小型猪特异性麻醉颉颃剂催醒作用可能与激活大脑皮层和海马等脑区的Na+,K+-ATP酶活性相关.     

Several physiologic aspects of the reactions of foals in the first three days after weaning

Changes have been observed in nine foals, six months of age, after weaning for the time of three days, namely in the intake of drinking water, hematocrit, sedimentation of erythrocytes and three minerals Na+, K+ and Cl-. Foals respond to weaning immediately, lowering significantly their consumption of drinking water and increasing high significance of Na+ and Cl-. Changes of level K+, hematocrit and sedimentation don't show any significant variations. The results reached are to be considered as an introductory information for further inquiry on the more extensive methodical basis.     

Exercise-induced hyperkalemia in hypothyroid dogs

We investigated the effect of hypothyroidism in dogs on (1) the Na+-, K+ -ATPase concentration in skeletal muscle, and (2) potassium (K+) homeostasis at rest and during exercise. Prior to and 1 year after induction of hypothyroidism by surgery and subsequent radiothyroidectomy, the Na+-, K+ -ATPase concentrations were quantified in biopsies of sternothyroid muscles of seven Beagle dogs by measuring [3H]ouabain binding capacity. In addition, plasma K+ concentrations were measured at rest and after treadmill exercise in six hypothyroid and seven euthyroid Beagle dogs. During hypothyroidism, the mean Na+ -, K+ -ATPase concentration in muscle biopsies was 41% lower than during euthyroidism. The mean resting plasma K+ value of the hypothyroid dogs was significantly (14%) higher than that of the euthyroid dogs. In the hypothyroid dogs, plasma K+ concentration increased significantly during exercise, whereas there was no rise in the euthyroid dogs. The rise in plasma K+ concentration could not be ascribed to muscle damage, as plasma creatine kinase concentrations remained within reference range. Also renal K+ retention was an unlikely explanation, as plasma aldosterone concentration and plasma renin activity rather increased than decreased during exercise. In conclusion, hypothyroid dogs tend to develop hyperkalemia during exercise, which for a large part can be explained by the severe reduction of the Na+ -, K+ -ATPase capacity in the skeletal muscle pool.     

Specificity and reversibility of the training effects on the concentration of Na+,K+-Atpase in foal skeletal muscle

The purpose of the present study was to determine whether training and detraining affect the Na+,K+-ATPase concentration in horse skeletal muscles, and whether these effects are specific for the muscles involved in the training programme. Twenty-four Dutch Warmblood foals age 7 days were assigned randomly to 3 groups: Box (box-rest without training), Training (box-rest with training: short-sprint) and Pasture (pasture without training). Exercise regimens were carried out for 5 months and were followed by 6 months of detraining. Five of the foals in each group were subjected to euthanasia at age 5 months and the remaining foals at 11 months. Muscle samples were collected from the deep part of the gluteus medius, semitendinosus and masseter muscles. The Na+,K+-ATPase concentration was quantified by [3H]ouabain binding. In the Training group, the concentration of Na+,K+-ATPase in gluteus medius and semitendinosus muscle, but not in masseter muscle, showed a relative increase of 20% (P<0.05) as compared to Box foals. After detraining for the subsequent 6 months, the concentration of Na+,K+-ATPase in semitendinosus muscle remained the same, while that in gluteus medius muscle was reduced by 10%. It is concluded that: 1) short-sprint training for 5 months induced an increase of the Na+,K+-ATPase concentration in gluteus medius and semitendinosus muscles of the foal. Interestingly, this effect persisted during the 6 months of the detraining period. Whether the higher Na+,K+-ATPase concentration due to training of young foals leads to a better athletic performance when they become mature still needs to be established; 2) the factors that initiate an increase in Na+,K+-ATPase concentration following training are likely to be located in the muscle itself and 3) the training effect may last for several months after returning to normal activity, especially in muscles containing a high percentage of fast-twitch fibres.     

Energy metabolism in canine erythrocytes associated with inherited high Na+- and K+-stimulated adenosine triphosphatase activity

Energy metabolism in canine erythrocytes associated with inherited high Na+- and K+-stimulated adenosine triphosphatase [(Na,K)-ATPase] activity (HK cells) was compared with that in normal canine erythrocytes (LK cells). Activities of some of the glycolytic enzymes in the HK cells were significantly higher than those in LK cells. The concentrations of adenosine triphosphate (ATP) and glycolytic intermediates in HK cells were almost equal to those in LK cells. Glucose utilization and lactate production by HK cells in vitro and incorporation of [32P]orthophosphate or [14C]glucose into 2,3-diphosphoglycerate in HK cells were higher than in LK cells. Radioactivity of [32P]ATP in HK cells was lower than in LK cells, but increased to approximately that of LK cells when (Na,K)-ATPase of HK cells was completely blocked by ouabain. When HK cells and LK cells were incubated in the absence of glucose, the concentration of ATP in HK cells was decreased more than that of LK cells. Although ouabain reduced the rate of decrease in ATP in HK cells, the decrease in ATP in HK cells was still 2-fold that in LK cells. The half-life of HK cells was about one-half that of LK cells. The results indicated that glycolysis is greater in HK cells than in LK cells, and that the increased glycolysis in HK cells was stimulated by an increased rate of ATP breakdown for active cation transport by the (Na,K)-ATPase and by increased degradation of ATP for some other pathway, eg, glutathione synthesis. Thus, the increased demand for ATP in HK cells might result in shortening the lifespan of HK erythrocytes.     

NaCl胁迫对苜蓿幼苗生长及不同器官中盐离子分布的影响

研究了不同浓度NaCl胁迫下(盐浓度处理为0,0.1%,0.3%,0.5%,0.7%,0.9%)苜蓿不同器官的生长情况(存活率、根长、茎长、叶片数),以及不同器官中Na⁺、K⁺、Ca²⁺、Mg²⁺的含量,并分析了K/Na、Ca/Na、Mg/Na值的变化。结果表明,存活率、根长、茎长与盐浓度呈负相关,且当NaCl≤0.1%时,有利于苜蓿的生长。除根茎叶中Na⁺含量、根中Ca²⁺与盐浓度呈正相关,茎叶中的Ca²⁺与盐浓度相关性不明显外,根茎叶中K⁺、Mg²⁺、K/Na、Ca/Na、Mg/Na均随盐浓度增加而显著下降,且盐处理与对照差异显著(P＜0.05)。另外从离子角度讨论了盐对苜蓿的毒害以及耐盐的原因。     

A proposed mechanism of monensin action in inhibiting ruminal bacterial growth: effects on ion flux and protonmotive force

Streptococcus bovis, a Gram-positive ruminal bacterium, was unable to grow in the presence of monensin. When monensin (5 mg/liter) was added to actively growing cultures, there was an immediate decrease in growth rate, and within 3 h no further growth was observed. Glucose utilization and lactate production continued for another 8 h even though growth had ceased. Monensin caused a decrease (P less than .05) in intracellular K+, a decrease (P less than .05) in intracellular pH and an increase (P less than .05) in intracellular Na+. The net exchange of K+ for Na+ and H+ via monensin was driven by the difference in concentration of K+ and Na+ across the cell membrane. Non-treated cells maintained a 70-fold gradient (inside higher) for K+, while the Na+ gradient was only 2.7-fold (inside higher). Previous models were based on a reverse mechanism whereby monensin would drive an efflux of Na+ from ruminal bacteria, and were not supported by experimental measurements of intracellular Na+ and K+.     

Expression of gastric ghrelin and H(+)-K(+)-ATPase mRNA in weanling piglets and effect of ghrelin on H(+)-K(+)-ATPase expression and activity in gastric mucosal cells in vitro

This study was designed to investigate the effect of ghrelin on gastric acid secretion in weaning piglets both in vivo and in vitro. Thirty newborn piglets were selected from six litters and on 28, 35 (weaning), 38, 42 and 45d of age, respectively, one piglet from each litter was killed and the mucosal tissue from gastric fundus was collected for detecting ghrelin mRNA as well as H(+)-K(+)-ATPase mRNA expression and activity. Primary cultures of gastric mucosal cells from 5-week-old weaning piglets were challenged with 3x10(-5), 3x10(-4), 3x10(-3), 3x10(-2) and 3x10(-1)nmol/ml h-ghrelin, respectively, for 4h in order to further clarify the effect of ghrelin on gastric H(+)-K(+)-ATPase mRNA expression and activity. Ghrelin mRNA expression in gastric fundus kept stable from 28d to 42d, followed by a sudden increase on 45d, exhibiting a peak that was significantly higher than any other age groups investigated. H(+)-K(+)-ATPase activity and mRNA expression showed similar trends of increase with slightly different timing. H(+)-K(+)-ATPase mRNA expression tended to increase on 42d, while H(+)-K(+)-ATPase activity started to rise from 35d, but neither of them reached significantly higher levels until 45d. In vitro, ghrelin significantly increased H(+)-K(+)-ATPase activity of gastric mucosal cells at 3x10(-4), 3x10(-3), and 3x10(-2)nmol/ml, but augmented H(+)-K(+)-ATPase mRNA expression only at 3x10(-4)nmol/ml. The results indicate that ghrelin mRNA expression is up-regulated 10 days after weaning in the gastric fundus of piglets, coinciding with the increase of H(+)-K(+)-ATPase mRNA expression and activity. Ghrelin acts on gastric mucosal cells to stimulate both mRNA expression and activity of H(+)-K(+)-ATPase in vitro.