Some enzyme interrelationships in serum and organs of Egyptian camel

In 7 normal healthy Egyptian one-humped camels aged 3-4 years, the relationships were studied between enzyme activities of lactic dehydrogenase (LDH), creatine phosphokinase (CPK), alkaline phosphatase (ALP), acid phosphatase (ACP), and cholonesterase (CHE) of serum and organs as well as between ACP and ALP and between LDH and CPK. Liver, heart, kidney, and spleen tissue samples as well as serum were analysed for total enzyme activity. The following results were obtained: --The heart was the organ with the highest activities of ALP, LDH, and CPK, and it contained high values of CHE, whereas the lowest activities of all enzymes were recorded from serum. The spleen exhibited of the highest activity of ACP. --Each of the serum enzymes ALP, LDH, and CHE were in strong inverse relationship with the corresponding enzymes in the liver. Strong inverse relationships existed also between serum LDH and kidney LDH as well as between CPK in serum and heart. --Direct relationships were remarkable serum LDH and that of spleen and heart as well as between serum ACP and that of liver and heart. --Interrelationships were inverse between ACP and ALP in liver, kidney, and heart, but weak direct interrelationships were characteristic between the 2 enzymes in serum and spleen. --LDH was inversely interrelated with CPK in serum and heart.     

Lactate dehydrogenase and creatine phosphokinase isoenzymes in tissues of laboratory animals

The lactate dehydrogenase (LDH) and creatine phosphokinase (CPK) isoenzyme distributions in tissues of the ICR mouse, Wistar rat, guinea pig and golden hamster were analyzed by histoelectrophoresis. Tissues obtained were as follows: liver, pancreas, stomach, small intestine, heart, femoral muscle, uterus, kidney, spleen, lymph node, cerebrum, spinal cord and erythrocyte. Histoelectrophoresis was for the direct analysis of LDH and CPK isoenzymes in the tissues and had high practical value compared with previous tissue-extraction methods. In tissues of the mouse, guinea pig and golden hamster, LDH isoenzymes showed five bands. In the rat, LDH isoenzyme was separated into four fractions. CPK isoenzyme showed three bands; BB, MB and MM. In some tissues, the MM band was separated into two sub fractions.     

Lactate dehydrogenase and its isoenzymes in the tissues and sera of clinically normal dogs

The total activity of lactate dehydrogenase (LDH) and the percentage distribution of its isoenzymes in the tissues and sera of clinically normal adult dogs are presented. Total LDH activity was greatest in skeletal muscle followed by heart muscle, kidney, small intestinal mucosa, liver, lung, pancreas and bone. Each tissue had a unique isoenzyme pattern and the proportions of the isoenzymes in serum suggested that liver is the source of normal serum LDH. The tissue isoenzyme patterns were similar to those obtained by other authors in human beings, horses, cattle, sheep and cats although in liver, differences between ruminants and monogastric animals including dogs were evident. The data presented provide a basis for the interpretation of serum LDH isoenzyme patterns in canine disease.     

Heat stability of serum lactate dehydrogenase and its isoenzymes in young and adult cattle and sheep. Evaluation of a relative heat stability test and serum determination of alpha-hydroxybutyrate dehydrogenase in diagnostic work

The heat stability of lactate dehydrogenase (LDH) has been investigated in serum from young and adult cattle and sheep. The thermoresistance of the isoenzymes was determined by electrophoresis of serum samples preincubated at different temperatures. Marked differences were found in the percentage distribution of isoenzymes in serum from the two species as well as in the heat stability. LDH in serum from sheep was inactivated at a lower temperature than that in serum from cattle, and inactivation occurred at a lower temperature in young than in adult animals. The enzyme was in both species less tolerant to elevated temperatures than what is reported for human serum. Procedures worked out for a so-called relative heat stability test of LDH in human clinical diagnosis may therefore give misleading results if they were applied uncritically to sera from these animals.The LDH isoenzyme pattern of some main organs in calves and sheep indicates that a serum heat stability test may be useful in the diagnosis of skeletal muscle injuries in sheep. In cattle the tissue isoenzyme distribution is assumed to be too uniform to give information about specific organ lesions either by serum electrophoresis or by a heating technique.In contrast to what has been reported in man, serum levels of α-hydroxybutyrate dehydrogenase (HBD) in cattle and sheep, as earlier reported in swine, are found to be far better correlated to total LDH than to the most thermostable isoenzyme, LDH₁.     

Activities of enzymes in the malate-aspartate shuttle and the isoenzyme pattern of lactate dehydrogenase in plasma and peripheral leukocytes of lactating Holstein cows and riding horses

The activities of the enzymes involved in the malate-aspartate shuttle and lactate dehydrogenase (LDH) and the pattern of the isoenzymes of LDH were determined in plasma and peripheral leukocytes of lactating Holstein cows and thoroughbred riding horses as representative herbivorous animals. In the horse plasma, LDH activities were significantly lower and AST activities were significantly higher than those in the cow plasma. The specific activities of cytosolic malate dehydrogenase (MDH), LDH and AST in the horse leukocytes were higher than those in the cows. The cytosolic ratio of MDH/LDH activity (ML ratio) in the horse leukocytes was significantly lower than that in the cow leukocytes owing to significantly higher activities of LDH. The ML ratio was considered to reflect the difference in energy metabolism in leukocytes between cows and horses. The plasma LDH isoenzyme patterns of cow and horse showed the characteristic as herbivorous animals with dominance of LDH-1, -2 and -3. The LDH isoenzyme patterns with dominance of LDH-3 and -4 in the horse leukocytes were remarkably different from those in the cow leukocytes. There were significant differences in activities of malate-aspartate shuttle enzymes, ML ratio and LDH isoenzyme patterns in the cytosolic fractions of leukocytes between the lactating cows and the riding horses.     

CLINICAL APPLICATIONS OF SERUM ENZYME DETERMINATIONS

This study determined the normal enzyme levels in horses, cattle, and dogs for a group of 10 enzymes. The enzyme tests were then evaluated under clinical conditions, and case histories are presented to illustrate the useful applications of these tests. Following clinical evaluations, the tests were grouped into profiles for the study of disease syndromes. Each profile contained the tests of greatest value in evaluating a specific organ or tissue. Sorbitol dehydrogenase (SDH) and glutamic dehydrogenase (GLDH) are enzymes found primarily in the liver and are combined with the less tissue specific glutamic oxalacetic transaminase (GOT) to monitor hepatocellular degeneration. Aldolase (ALD) and creatine phosphokinase (CPK) are enzymes found primarily in skeletal muscle. Elevations of ALD, CPK, and GOT can assist in the diagnosis of skeletal muscle degeneration. Not only are the newer enzyme tests relatively organ specific but also offer greater sensitivity than has previously been obtained.     

Alkaline phosphatase and its isoenzymes in the tissues and sera of normal dogs

The total alkaline phosphatase (AP) activity and the pattern of its isoenzymes were studied in the tissues and sera of normal adult dogs. Small intestine mucosa showed the greatest total AP activity followed by kidney, bone, pancreas, liver, lung, skeletal muscle and heart muscle. After separation by agarose gel electrophoresis, each tissue showed only one isoenzyme except lung which showed two. The tissue isoenzymes, in decreasing order of migration distance towards the anode, were as follows: fast lung isoenzyme, liver or slow lung isoenzyme, the group consisting of skeletal muscle, bone, small intestine and pancreas isoenzymes and, finally, the kidney isoenzyme. Two isoenzymes occurred in serum. The major band corresponded to liver and the slow lung isoenzyme, while the minor band was considered to be the corticosteroid-induced isoenzyme, previously thought to be absent from normal serum.The AP isoenzyme patterns in lung and skeletal muscle and the presence of an isoenzyme migrating an identical distance to the corticosteroid-induced isoenzyme do not appear to have been reported before in normal dogs.     

Alkaline phosphatase and its isoenzymes in the tissues and sera of normal dogs

The total alkaline phosphatase (AP) activity and the pattern of its isoenzymes were studied in the tissues and sera of normal adult dogs. Small intestine mucosa showed the greatest total AP activity followed by kidney, bone, pancreas, liver, lung, skeletal muscle and heart muscle. After separation by agarose gel electrophoresis, each tissue showed only one isoenzyme except lung which showed two. The tissue isoenzymes, in decreasing order of migration distance towards the anode, were as follows: fast lung isoenzyme, liver or slow lung isoenzyme, the group consisting of skeletal muscle, bone, small intestine and pancreas isoenzymes and, finally, the kidney isoenzyme. Two isoenzymes occurred in serum. The major band corresponded to liver and the slow lung isoenzyme, while the minor band was considered to be the corticosteroid-induced isoenzyme, previously thought to be absent from normal serum. The AP isoenzyme patterns in lung and skeletal muscle and the presence of an isoenzyme migrating an identical distance to the corticosteroid-induced isoenzyme do not appear to have been reported before in normal dogs.     

罗斯及印第安河鸡发育时期组织LDH同工酶变化

罗斯及印第安河鸡在胚胎第3d的混合胚中出现LDH_(1～3)3种同工酶(3条区带,梯度凝胶电泳),或LDH_(1～4)4种同工酶(6～7条区带,等电聚焦电泳);第7d检出心肌、胸肌和脑的5种同工酶(LDH_(1～5),5～7条区带);第12d检出肝脏的LDH_(1～5)5种同工酶(8或15条区带).组织LDH同工酶区带数随发育过程明显增多,同种鸡同一组织不同发育时期的LDH同工酶活性变化相差显著或极显著.在相同发育时期,印第安河鸡胸肌和肝脏的LDH_1相对活性均明显高于罗斯鸡(P<0.01).两种鸡心肌、胸肌、肝和脑组织LDH同工酶的等电点主要分布在pH6.0～8.2内.     

Value of measurements of the serum activities of creatine phosphokinase,aspartate aminotransferase and lactate dehydrogenase for predicting whether recumbent dairy cows will recover

Over a period of seven years, 262 recumbent dairy cows were investigated, and serum samples were analysed for creatine phosphokinase (CPK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) activities. The results from cows for which all three measurements were available, were analysed statistically by the non-parametric receiver operating characteristic (ROC) method, to evaluate the value of the serum enzyme activities for predicting a failure to recover. The sensitivity and specificity of the activity levels were calculated over all cut-off points, and ROC curves were created by plotting sensitivity as a function of 1--specificity at each cut-off point for samples grouped by the interval after the cows became recumbent. The predictive values of the tests were compared by calculating the areas under the curves, and the positive and negative predictive values of the tests were calculated and plotted against the prevalence of a failure to recover. The results from each of the enzymes were significantly better than from a random test with no discriminatory ability. The optimal cut-off points maximising the sensitivity and specificity of the tests were 2330, 2225 and 171 U/litre for CPK, LDH and AST, respectively. The predictive value of AST was significantly better than that of CPK or LDH, and measures made on the second and third day of recumbency were significantly better with optimal cut-off points of 128 and 189 U/litre, respectively.     

Effects of physical stress on serum enzymes and other blood constituents in sheep

In connection with transfer of sheep from the lowland near Oslo to mountain pastures at an altitude of 1,200 m above sea level, investigations were carried out in 37 animals to study the effect of physical stress on serum enzymes and other blood constituents. The sheep were adult ewes and lambs. About half of the animals had been accustomed to outdoor life on pasture for more than one month, while the others were moved directly from indoor feeding. Blood was collected before departure, after six hrs. of long-distance transportation by lorry, and after three hrs. of subsequent continuous herding on foot. The following blood components were determined: Aspartate aminotransferase (AspAT = GOT), alanine aminotransferase (A1AT = GPT), α-hydroxybutyrate dehydrogenase (HBD), total lactate dehydrogenase (LDH), LDH isoenzymes, alkaline phosphatase, calcium, inorganic phosphorus, magnesium, blood sugar, total serum proteins, and haemoglobin.In summary, it may be said that the lambs reacted with greater changes of the blood components than adult animals, and that untrained, indoor fed lambs were distinctly more sensitive than those taken from pasture. The “indoor” lambs showed a statistical significant increase from the initial values in AspAT, HBD, total LDH, the isoenzymes LDH₃ and LDH₄, and blood sugar. Significantly decreased values were recorded in Ga, P, Mg, and total serum protein. Some of these changes, as in Mg and P, were most pronounced after transportation, while elevations of serum enzyme levels continued to increase during the subsequent herding.Based upon the shift in LDH isoenzyme distribution towards a more cathodically dominated pattern it is supposed that the main origin of increased serum enzyme activity was skeletal muscle.     

The mechanisms relating to increases in plasma enzymes and isoenzymes in diseases of animals

Changes in plasma or serum enzymes and isoenzymes are useful indicators of tissue damage in many diseases. Enzyme increases are usually related to leakage of enzymes from damaged cells. The amount of increase depends on factors such as the concentration of enzyme in the cells, the rate of leakage of enzyme from cells and the rate of clearance of enzyme from plasma. There are important species variations in the tissue concentrations and plasma clearance rates of enzymes and isoenzymes.     

The Comparative Morphology of the Muscle Tissues and Changes of Constituents in the Pig Types

The authors have revealed the main value characteristics of pork production by testing in five different types of pig the volume of contractile and collagen proteins, that of proteoglycans, the constituents of blood and the enzymes of the blood plasma. The contractile proteins of the muscle tissues basically determine the quality of pork. The same applies to the water retention capacity, colloidal characteristics and glycogen content of meat. The amount of contractile proteins has decreased in the best meat producing types of pig. Parallel with the decrease of white meat, and with the increase in the volume of ham, chop and chuck the contractile protein content of muscles decreased. The scientific fact according to which there is a certain correlation among the changes in the volume of contractile proteins, blood sugar level, blood serum CPK and the intensity of activity of the LDH enzymes promotes the qualifying of live animals and the work of the geneticists aiming at the increasing of the contractile protein content of the muscle tissues of pigs by selection. According to tests carried out by us the primary cause of PSE changes is a decreased volume of contractile proteins. Increased stress sensitivity and all the other factors have but a secondary importance and are all consequential. The decrease in the quantity of contractile proteins or--it is better to put it this way--the lack of the proper amount of such proteins characterizing a fully developed pig's organism is caused by the nowadays usual breeding technologies and can be well explained by those selection activities which aim at a one-sided kind of pork production.     

三种药物对猪棘头虫LDH同工酶的抑制试验

根据猪棘头虫与宿主（猪）血清的ＬＤＨ同工酶的差异，用０．１％硝硫氰醚、０．１％丙硫咪唑和０．１％吡喹酮分别对猪棘头虫雄虫、雌虫和猪血清的ＬＤＨ同工酶进行了抑制试验，结果除硝硫氰醚对蛭形巨吻棘头虫雄虫、雌虫的ＬＤＨ同工酶有抑制作用外，其余的试验组与对照组均无差异。硝硫氰醚对雄虫ＬＤＨ同工酶的第２、３、４带和雌虫ＬＤＨ同工酶的第２、３带有不同程度的抑制作用，特别是雌虫ＬＤＨ同工酶的第２带最敏感，在０．１％的浓度下，该酶带被完全抑制，即没显示出来，硝硫氰醚对雄虫和雌虫ＬＤＨ同工酶的极弱带（它们各自的第１带）均无抑制作用；临床实验也显示出硝硫氰醚对治疗猪棘头虫病有一定疗效，这些表明硝硫氰醚抑制蛭形巨吻棘头虫的ＬＤＨ同工酶活力可能是该药物对该虫体的作用机理之一。     

Lactate dehydrogenase and creatine kinase isoenzyme levels in the tissues and serum of normal lambs

The normal activities of lactate dehydrogenase and creatine kinase isoenzymes in the tissues and serum of clinically normal five-and-a-half-month-old lambs are presented. The percentage distribution of the lactate dehydrogenase isoenzymes in serum, liver, heart, lung and skeletal muscle were in agreement with previous studies but the distribution in ovine abomasum, small intestine, large intestine and red blood cells has not been previously described. Up to five creatine kinase isoenzymes were detected in ovine tissues and four in serum. One creatine kinase isoenzyme present in serum and tissues was thought to represent a mitochondrial isoenzyme which is absent from the normal serum of other species. Estimations of both lactate dehydrogenase and creatine kinase isoenzymes allowed all the tissues examined to be distinguished and were therefore more likely to allow tissue-specific isoenzyme patterns to be detected in serum than the estimation of the isoenzymes of either enzyme alone.     

Enzymes Sériques pour le Diagnostic de la Nécrose Hépatique Aigüe Expérimentale

The activity of certain serum enzymes ornithine carbamyl transferase (OCT), serum isocitric dehydrogenase (SIC-D), total serum lactic dehydrogenase (LDH) and its isoenzymes (LD₁ and LD₅) was evaluated as a mean of assessing experimental hepatic necrosis on dogs treated with CCl₄. Measurement of activity levels of these enzymes, seldom carried out in veterinary clinical pathology, was made together with tests commonly used in our laboratories: serum glutamic pyruvic transaminase (SGPT), serum glutamic oxalacetic transaminase (SGOT) and serum alkaline phosphatase (SAP), cholesterol, bilirubin and prothrombin time. Measurement of the level of OCT was useful in the diagnosis of liver necrosis. The SIC-D level was important during the first four days of the experiment, but on subsequent days, the enzymatic activity was practically normal. Because of the wide variations of LDH serum levels in normal animals and since many factors influence its activity, the measurement of this enzyme and its isoenzymes was not a good index in the diagnosis of canine liver necrosis. The evaluation of cholesterol and bilirubin was judged of secondary importance because these metabolites are not specific to hepatic problems.     

绵羊组织器官LDH同工酶特异性研究

本研究利用不连续聚丙烯酰胺凝胶电泳法对绵羊的８种组织器官ＬＤＨ同工酶的分布及相对活性进行了分析测定。比较ＬＤＨ同工酶的酶谱特征。结果表明，绵羊的ＬＤＨ同工酶分布特征不同于其它动物，其相对活性有显著差异。骨骼肌中ＬＤＨ５活性比ＬＤＨ１强，Ｍ亚基比例大于Ｈ亚基；肝脏和心脏中ＬＤＨ１的活性最强。     

复方丁氨丙磷溶液的抗热应激作用及其机理研究

为探讨复方丁氨丙磷溶液的抗热应激作用及作用机理,以黄羽肉鸡为对象,检测各处理组肉鸡生产性能,体温及血清T3、T4、胰岛素和皮质醇等内分泌激素,血清肌酸磷酸激酶、碱性磷酸酶、乳酸脱氢酶和血糖等生化指标.将45只1日龄肉鸡仔鸡随机分成3组,每组15只,分别给予生理盐水和不同剂量的复方丁氨丙磷溶液.于14日龄开始热暴露,在高温条件下继续饲养2周,并观察和检测各项指标.试验结果表明,复方丁氨丙磷溶液能明显降低热应激肉仔鸡直肠温度,显著提高T3/T4值和胰岛素含量,能极显著降低血清肌酸磷酸激酶、碱性磷酸酶和乳酸脱氢酶活性,同时提高血糖含量,而对血清皮质醇含量无显著影响.因此,复方丁氨丙磷溶液可通过调整肉仔鸡血清部分激素含量和改变血清部分生化指标来发挥其抗应激作用.     

Lactate dehydrogenase isoenzymes in the lungs of sheep with acute and chronic pneumonia

Total lactate dehydrogenase and the absolute and percentage levels of its isoenzymes were measured in lung lesions and macroscopically normal areas of lung from lambs with chronic proliferative exudative pneumonia and acute pasteurella pneumonia. Lung lesions had a higher total enzyme activity which was associated mainly with increases in the activity of the LDH4 and LDH5 isoenzymes, particularly in chronic pneumonia, and gave lung lesions a considerable potential for altering the serum isoenzyme distribution. Thus, the nature of any changes in the serum isoenzyme distribution will depend on whether the isoenzymes are released from abnormal or normal areas of lung. This appears to be the first report on lactate dehydrogenase isoenzymes in ovine pneumonia.