Changes in activities of enzymes related to energy metabolism in canine lymphoma cells

Alterations in the activities of enzymes related to energy metabolism in canine lymphoma cells were investigated. Cytosolic pyruvate kinase (PK) and mitochondrial malate dehydrogenase (MDH) activities in lymphoma cells were significantly higher than those in lymphocytes obtained from lymph nodes of healthy dogs, whereas cytosolic lactate dehydrogenase (LDH) activity was significantly lower in lymphoma cells. The cytosolic M/L ratio (MDH activity/LDH activity), which is considered to be a good indicator of energy metabolism related to glucose utilization in animal tissues, was significantly higher in lymphoma cells than in the normal lymphocytes.     

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.     

营养限制与补偿对蒙古育成羯羊肝脏关键酶活性的影响

本试验旨在阐明营养限制与补偿对蒙古育成羯羊肝脏关键酶活性的影响。试验选用40只平均体重为(14.72±1.10)kg的3月龄蒙古育成羯羊,随机分为对照组(CG)和双限组(RG)。试验分营养限制期(60d)和营养补偿期(90d)。限制期2组饲粮对应能氮水平为代谢能(ME):10.88(对照组)和8.62(双限组)MJ/kg;粗蛋白质(CP):15.0%(对照组)和5.7%(双限组)。补偿期各组饲喂同一能氮水平(ME:9.75 MJ/kg;CP:12.0%)的饲粮。在限制期与补偿期末分别测定蒙古育成羯羊肝脏中己糖激酶(HK)、丙酮酸激酶(PK)、乳酸脱氢酶(LDH)、果糖-1,6-二磷酸酶(FBP1)、苹果酸脱氢酶(MDH)、α-磷酸甘油脱氢酶(GAPDH)、丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)和谷氨酸脱氢酶(GLDH)的活性。结果显示:限制期结束时双限组蒙古育成羯羊肝脏中HK、ALT、AST、GLDH活性显著低于对照组(P<0.05),而PK、LDH、FBP1、MDH、GAPDH活性显著高于对照组(P<0.05);经过90d补偿后,双限组蒙古育成羯羊肝脏中HK、PK、MDH、ALT活性显著高于对照组(P<0.05),而LDH、FBP1、GAPDH、AST、GLDH活性与对照组差异不显著(P>0.05)。结果提示,限制期双限组蒙古育成羯羊糖酵解途径关键酶活性降低,糖异生途径关键酶活性升高,脂肪分解途径关键酶活性升高,这与机体在低能氮营养水平时脂肪动员加强有关,蛋白质代谢途径关键酶活性降低则表明限制期蛋白质代谢降低;补偿期双限组蒙古育成羯羊总体糖酵解、糖异生和脂肪分解途径关键酶活性基本恢复到对照组水平,而PK活性显著提高,脂肪合成和蛋白质代谢途径关键酶活性升高可能与此期较高的生长速率相关。     

Activities of enzymes in some types of peripheral leucocytes may reflect the differences in nutrient metabolism between dogs and cats

Plasma metabolites and immunoreactive insulin (IRI) concentrations and enzyme activities of some types of peripheral leucocytes were measured to clarify one aspect of the differences in nutrient metabolism between dogs and cats. There were no significant differences in plasma concentrations of glucose, triglyceride, free fatty acids and IRI between dogs and cats. Higher total cholesterol and lower HDL cholesterol concentrations were observed in feline plasma, and H/T ratio (HDL/total cholesterol concentrations) was significantly lower than that in canine plasma. The cytosolic activities of fructokinase (FK), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G6PD) and lactate dehydrogenase (LDH) were significantly higher and the activities of cytosolic malate dehydrogenase (MDH) and mitochondrial glutamate dehydrogenase (GLDH) were significantly lower in feline leucocytes than those in canine leucocytes. Higher activities of FK, PK and G6PD, which regulate the rate of biosynthesis of fatty acids, may reflect the different characteristics in nutrient metabolism in feline tissues from canine tissues.     

Serum enzyme activity of newborn calves, pigs and lambs

Serum activities of aspartate aminotransferase (AspAT = GOT), alanine aminotransferase (AlAT = GPT), and total lactate dehydrogenase (LDH) have been investigated in newborn calves, pigs, and lambs. In the two latter species the LDH isoenzyme distribution in serum was also studied. Blood samples were taken at frequent intervals from birth to 48–72 hrs. post partum.Calves and pigs were born with very low serum enzyme values, whereas lambs showed a picture more similar to what has been reported in human infants. In all species a marked temporary enzyme increase occurred during the first 24–48 hrs. This elevation was found not to be due to colostrum feeding, since a parallel increase was found in starved animals. Possible regulating mechanisms are discussed.The LDH isoenzyme pattern proved to be more stable than total LDH in the early post-natal period.The percentage isoenzyme distribution, however, showed characteristic differences from that found in adult animals of the same species.     

Acute immobilization stress in prone position in susceptible Pietrain and resistant Duroc pigs. II. Glycolytic metabolism in Piettrain and Duroc skeletal muscle and liver

Duroc (D) and Pietrain (P) pigs were used to study the effect of acute immobilization stress on carbohydrate metabolism. Changes in the activities of several glycolytic enzymes and the content of glycogen in longissimus dorsi muscle (LDM), glycogen content in the liver and glucose level in plasma were observed. The glycogen level in LDM of pigs was about five times higher in P pigs compared with D ones. The immobilization stress of 15 min caused a decrease of glycogen LDM content by 27% in D and by 44% in P pigs. In the liver, these changes were 75% in P pigs, but in D pigs the glycogen content was lowered during the time of experiment by 37% (control versus 60 min stress). Plasma glucose level was elevated by 118% in D and by 50.5% in P pigs after 15 min of immobilization stress. The activities of the glycolytic enzymes investigated changed in different way in LDM: the PGluM decreased by 20% in D and by 19% in P pigs; the GPI by 8.5% in both strains of pigs; the PFK and PK were higher in D than in P pigs (some statistically significant differences between PK activities of control and 15‐, 30‐ and 60‐min‐stressed animals); the ALD activity was similar in both investigated strains of pigs and LDH was lower in D than in P pigs. It can be assumed that the changes led to the new homeostasis connected with the stress to which the pigs were subjected.     

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.     

Changes of serum enzyme activities in cows with milk fat depression

Serum activities of sorbitol dehydrogenase (SDH), lactate dehydrogenase (LDH), gamma-glutamyltranspeptidase (gamma-GT), and alkaline phosphatase (ALP) were measured before, during, and after milk fat depression in 9 cows fed a high-concentrate ration during 2 experiments. In 7 of the 9 cows, increases in serum SDH and LDH activities were observed during milk fat depression. The gamma-GT activity showed only moderate changes, whereas the ALP activity remained unchanged. During recovery from milk fat depression, decreases of SDH, LDH, and gamma-GT activities were found in nearly all cows. Association of this phenomenon with changes found in beef cattle, such as ruminal lesions and liver abscesses, are discussed.     

Artifactually increased serum bicarbonate values in two horses and a calf with severe rhabdomyolysis

Extremely high bicarbonate (HCO3-) and anion gap values were measured in two horses and a calf using the Hitachi 911 automated serum biochemistry analyzer. All three animals had severe muscle disease as evidenced by markedly increased aspartate aminotransferase and creatine kinase activities. Laboratory error was suspected as the source of the increased HCO3- because values calculated from blood gas analysis were normal. It was hypothesized that increased serum lactate dehydrogenase (LDH) activity and pyruvate concentration overwhelmed the oxamate LDH inhibitor in the enzymatic HCO3- assay, resulting in consumption of NADH and falsely elevated spectrophotometric reading. Serum LDH activity was markedly increased in all three patients. In an attempt to reproduce this interference in vitro, LDH and pyruvate were added to normal bovine serum. Bicarbonate concentration was artifactually increased in a linear, dose-response relationship proportional to the amount of LDH activity in the sample; addition of pyruvate augmented this increase. It was concluded that increased serum LDH activity and pyruvate concentration secondary to severe muscle disease can result in artifactual increases in serum HCO3- values obtained by routine enzymatic assay.     

盐穗木醇提取物对家兔血清酶的影响

探索盐穗木对家兔血清酶的影响。选取临床健康、体重相近的家兔12只,根据灌喂剂量分为对照组、低剂量组、中剂量组、高剂量组,每组3只。在对家兔连续灌喂盐穗木醇提取物后,分别于7、14、21和28d采血、分离血清,测定血清中AST、ALT、ALP和LDH的浓度。结果显示,以0.9%生理盐水溶解盐穗木醇提取物,各试验组与对照组相比,AST活力均差异显著(P0.05);ALT活力无统计学上差异;ALP、LDH活力在各试验组与对照组之间存在显著或极显著差异。以40%乙醇溶解盐穗木醇提取物,高剂量组血清中AST、ALP活力及中剂量组LDH活力与对照组相比差异极显著(P0.01);但各试验组与对照组相比,ALT活力无统计学差异。随着盐穗木醇提取物灌喂浓度的提高,血清中AST、ALT、ALP和LDH活力呈剂量依赖关系,说明一定剂量的盐穗木醇提取物对家兔肝脏功能有一定影响。     

Influence of dietary tannic acid and polyethylene glycol on growth and intestinal D-xylose absorption of broiler cockerels and activity of serum enzymes

1. In an experiment on broiler cockerels, the influence of tannic acid (TA) and polyethylene glycol (PEG) on body weight gain (BWG), feed conversion ratio (FCR), weight of intestine and liver, the activities of serum enzymes LDH, AST, ALT and intestinal absorption function were investigated. 2. Broiler cockerels were given either a commercial diet alone (control group) or a commercial diet with TA (20 g/kg), PEG (10 g/kg) or TA plus PEG (20 + 10 g/kg), for 10 d. 3. On the last day of the experiment, all birds and remaining feed were weighed individually and a sample of blood was taken to measure the serum activities of lactate dehydrogenase (LDH), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). The capacity of intestinal cells for the absorption of D-xylose was measured. Finally all birds were killed humanely and the intestine and liver were weighed. 4. The results showed that TA significantly reduced BWG and FCR, as well as the activity of LDH, AST and ALT. 5. TA also increased the relative weight of the intestine. Adding PEG alone had no effect on any of the measured parameters. 6. However, PEG improved significantly BWG, FCR and the activity of LDH and AST of TA-fed birds. 7. The plasma D-xylose concentration of experimental birds was similar for all dietary treatments most likely because of temporal separation between feeding the dietary TA and administering the D-xylose. 8. It was concluded that the presence of tannins in the GI lumen of the bird was necessary to affect the processes involved in the absorption of simple sugars such as D-xylose, at the level of intestinal absorptive cells.     

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.     

Serum enzyme changes in newborn calves fed colostrum

In 10 newborn Holstein calves, which were initially bottle-fed on pooled colostrum, there were transient increases in several serum enzymes. Within a few hours, the mean serum gamma-glutamyl transferase (gammaGT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), acid phosphatase (ACP), and lactate dehydrogenase (LDH) activities increased 62.5, 2.4, 2.0, 1.8 and 1.3 fold, respectively. In two other calves given initial feeds of enzyme-free pasteurized milk, there were similar increases in serum enzymes, except serum gammaGT. Correct interpretation of serum enzyme activities in newborn calves must take into account the normal increases which occur after feeding colostrum. Serum gammaGT was the only enzyme to increase markedly as a result of its absorption from colostrum. The other serum enzymes are presumably released from the tissues of the calf. The increased activities of serum CK observed in some newborn calves probably resulted from trauma during birth or increased muscular activity after birth.     

Activity of some enzymes in the serum of dromedary camels.

The activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), acid phosphatase (ACP), lactate dehydrogenase (LDH) and isocitric dehydrogenase (ICD) in the serum of 60 healthy dromedary camels of either sex and different ages (one to 25 years) were determined. The results were analysed with respect to time of year (December-January and May-June), sex and age groups (below four years; four to 10 years; and over 10 years). The overall mean activities of AST, ALT, ALP, ACP, LDH and ICD were 36.1 +/- 0.35, 4.65 +/- 0.35, 27.21 +/- 0.43, 7.18 +/- 0.21, 479.0 +/- 7.33 and 7.74 +/- 0.17 iu litre-1, respectively. Activities of AST, ALT, ALP and ACP were significantly higher during extremely hot conditions (May-June) than in extreme cold (December-January) while the activity of LDH was higher in extremely cold conditions. Analysis of data based on sex revealed that AST, ALT and ALP activities in the serum of male animals were significantly higher than in female animals. The activities of all the enzymes were highest in animals under four years and then gradually decreased with age being lowest in the animals over 10 years.     

早期限饲对肉鸡心肌易颤性和血清心肌酶活性、电解质水平的影响

艾维因商品肉鸡于 4～ 12日龄时进行 10 %限饲 ,分别于 2 5、32日龄进行心肌易颤性的评价和血清心肌酶活性、血清电解质水平的检测。结果如下 :(1)早期限饲可以提高肉鸡的输钾诱颤阈 ,表明可降低肉鸡的心肌易颤性 ;(2 )早期限饲可以降低血清心肌酶 (AST、L DH、CK)的活性 ,表明可增强肉鸡的心肌机能 ;(3)早期限饲对血清电解质水平无明显影响。本试验验为进一步阐明早期限饲降低肉鸡猝死综合征发生的机制提供了理论依据     

Utilization of an enzyme heat stability test and erythrocyte glycolytic intermediate assays to assist in the diagnosis of canine pyruvate kinase deficiency

A macrocytic hypochromic anemia, with marked reliculocytosis and large numbers of circulating nucleated erythrocytes, was recognized in a 3 1/2-year-old male Beagle-cross dog. A presumptive diagnosis of erythrocyte pyruvate kinase (PK) deficiency was made, even though PK activity from the patient was within the range of activities measured for normal dogs, because the PK activity should have been several times normal in light of the large number of reticulocytes present. The PK activity in a hemolysate from the patient was heat-labile compared to activities in hemolysates from normal dogs, a finding consistent with results from a previous study of PK-deficient Basenji dogs. Seven phosphorylated glycolytic intermediates and 2,3-diphosphoglycerate were measured in protein-free extracts of erythrocytes from the patient. All were present in concentrations above that present in normal canine erythrocytes, further supporting the diagnosis of PK deficiency.     

Lactate dehydrogenase isoenzymes in blood and cerebrospinal fluid from healthy beagles

Serum and CSF lactate dehydrogenase (LDH) activity and LDH isoenzyme profile, as well as total protein, were measured in samples obtained from 26 healthy Beagles. The LDH activity in serum was approximately 10 times that in the CSF. The CSF LDH isoenzyme profile was a mirror image of the serum LDH isoenzyme profile. Age was negatively correlated (ie, as age increased, the activity decreased) with LDH3 in CSF and LDH5 in serum and was positively correlated with LDH1 in serum. Seemingly, simultaneous measurement of serum and CSF LDH activity and LDH isoenzyme profile can aid in establishing status of the blood-brain barrier, whether brain damage exists and whether other organ systems are involved.     

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.     

Plasma pyruvate kinase activity vs creatine kinase activity as an indicator of the porcine stress syndrome

Plasma pyruvate kinase (PK) and creatine kinase (CK) activities were increased significantly (P less than 0.001 and P less than 0.05, respectively) in homozygote halothane-reacting pigs (nn), compared with those activities in homozygote nonreacting pigs (NN). Pyruvate kinase activity was less variable within groups than was CK activity, allowing more effective discrimination between nn and NN geno-types. The PK and CK activities in plasma increase with age in halothane-reacting pigs and the nonreacting pigs. Enzyme activities in heterozygote (Nn) nonreacting pigs did not differ significantly (P greater than 0.05) from enzyme activities of homozygote (NN) nonreacting pigs. Although PK was better than CK in identifying stress-susceptible pigs, age-related effects and the failure to identify heterozygotes may restrict the use of plasma PK activity as a diagnostic test for the stress syndrome.     

猪带绦虫囊尾蚴体内发育过程中囊壁生化指标的变化

对猪体内发育30，40，60，80，95日猪囊尾蚴囊壁的已糖激酶（HK），丙酮酸激酶（PK），乳酸脱氢酶（LDH），苹果酸脱氢酶（MDH），葡萄糖－6－磷酸脱氢酶（G6PDH）活性和葡萄糖（GLc)，乳酸（LAC）含量进行测定，结果表明，HK，PK，MDH活性和GLc含量在60，80，95日高于或显著高于30，40日（P＞0．05，P＜0．05），说明随虫体的生长发育糖代谢逐渐增加。