Cytosolic ratio of malate dehyrogenase/lactate dehydrogenase activity in peripheral leukocytes of race horses with training

The activities of the enzymes involved in the malate-aspartate shuttle and m RNA expression of malate dehydrogenase (MDH), a crucial enzyme for the NADH shuttle that produces ATP in glucose metabolism in the peripheral leukocytes of horses, were measured to investigate the change in metabolic states with training. There were no significant differences in plasma glucose and immunoreactive insulin concentrations between race horses and riding horses, used as a comparable reference. The cytosolic and mitochondrial MDH activities in leukocytes of race horses were significantly higher than those of riding horses. High activities of MDH in leukocytes of race horses were confirmed by RT - PCR analysis on the total RNA extracted from the whole blood. The cytosolic ratio of MDH /lactate dehydrogenase (LDH) activity (M/L ratio) in leukocytes of race horses was significantly higher than in those of riding horses. Increase in the M/L ratio was considered to reflect elevation of energy metabolism in animal tissues. The M/L ratio may be a useful parameter to evaluate the difference in metabolic states between race horses and riding horses.     

Canine Leishmaniosis: An Italian Emergency

Activities of enzymes related to energy metabolism and isoenzyme patterns of lactate dehydrogenase (LDH) were determined in peripheral leukocytes and livers of Holstein dairy cows and Institute of Cancer Research (ICR) mice. In dairy cow liver, activities of enzymes in glycolysis, malate–aspartate shuttle and lipogenesis were lower, but activities of glucose-6-phosphatase in gluconeogenesis were higher than those in mouse liver. Glucokinase activities were below detection limit in leukocytes and liver of the cows. Dairy cow leukocytes and liver showed the isoenzyme patterns with dominance of LDH-1, -2 and-3, whereas mouse leukocytes and liver showedthat LDH-5 was dominant. The LDH isoenzyme patterns were very similar between leukocytes and liver in each animal species. Some enzymes in leukocytes may reflect those enzymes activities in liver and be a useful indicator for energy metabolism in animals.     

Comparison of enzyme activities in plasma and leukocytes in dairy and beef cattle

Concentrations of plasma glucose, immunoreactive insulin (IRI) and free fatty acid (FFA) and activities of enzymes related to energy metabolism and lactate dehydrogenase (LDH) isoenzyme pattern in plasma and leukocytes were investigated in lactating Holstein cows (dairy cattle) and fattening Japanese Black Wagyu x Holstein steers (beef cattle). IRI concentrations and LDH and malate dehydrogenase (MDH) activities in the plasma of beef cattle were significantly higher than those in dairy cattle. The cytosolic ratio of MDH/LDH activity in the leukocytes of beef cattle was significantly higher than that of dairy cattle. These findings might be associated with the different energy metabolism between dairy and beef cattle.     

Enzyme activities in some types of peripheral leukocytes of thoroughbred race horses before and after the races

Glucose, free fatty acid (FFA), triglyceride (TG) and immunoreactive insulin (IRI) concentrations in plasma and activities of enzymes related to energy metabolism in some types of peripheral leukocytes were measured in thoroughbred race horses before and after racing. Glucose, FFA, TG and IRI concentrations and enzyme activities did not change significantly in plasma. However, the activities of cytosolic hexokinase, malate dehydrogenase (MDH), mitochondrial MDH and aspartate aminotransferase decreased significantly in leukocytes of the horses after the races. The cytosolic ratio of MDH/LDH activity (ML ratio) in leukocytes decreased significantly after racing, and the ratio may be a useful indicator to evaluate energy metabolism in race horses.     

The Activity Ratio of the Cytosolic MDH/LDH and the Isoenzyme Pattern of LDH in the Peripheral Leukocytes of Dogs,Cats and Rabbits

The activities of malate dehydrogenase (MDH) and lactate dehydrogenase (LDH) and the pattern of the isoenzymes of LDH were determined in the peripheral blood leukocytes of dogs, rabbits and cats. Rabbits had significantly higher plasma glucose concentrations than dogs or cats. Feline leukocytes showed higher LDH and lower MDH activities than canine or rabbit leukocytes. The M/L ratio, defined as the MDH activity divided by the LDH activity in cytosolic fractions, was considered to be a good indicator with which to evaluate the metabolic state in animal tissues. The M/L ratio was highest in canine and lowest in feline leukocytes. LDH-2 and LDH-3 isoenzymes were dominant in canine leukocytes. LDH-1 and LDH-2 were dominant in rabbit leukocytes, whereas LDH-5 was dominant in feline leukocytes. It was evident that there were significant differences in energy metabolism between the leukocytes of dogs, rabbits and cats.     

Activities of Enzymes Related to the Malate–Aspartate Shuttle in the Blood Cells of Thoroughbred Horses Undergoing Training Exercise

The activities of the enzymes related to the malate–aspartate shuttle, which convert cytosolic NADH into mitochondrial NADH, were measured in red and white blood cells from thoroughbred horses undergoing continuous training (race horses) and compared with those in blood cells from riding horses. The activities of malate dehydrogenase (MDH), a rate-limiting enzyme for the malate-aspartate shuttle, were significantly elevated in the white blood cells (WBC) from race horses compared with those from riding horses. There were no significant differences in the activities of the enzymes in the red blood cells between race horses and riding horses. This increase in the MDH activity in their WBC is considered to reflect the increased metabolic activity in the race horses resulting from the training.     

Malate dehydrogenase activities are lower in some types of peripheral leucocytes of dogs and cats with type 1 diabetes mellitus

The activities of the enzymes in the malate-aspartate shuttle were measured in peripheral leucocytes of spontaneous type 1 diabetic dogs and cats treated with insulin injections. In the diabetic dogs and cats, fasting plasma glucose concentrations were three- or fourfold greater than the control levels in spite of insulin injections and the activities of cytosolic malate dehydrogenase (MDH), one of pivotal enzymes in the malate-aspartate shuttle, were remarkably lower than the controls. Depressed expression of cytosolic MDH mRNA was confirmed by RT-PCR analysis in the diabetic animals. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M / L ratio) in leucocytes of the diabetic animals was significantly lower than that of normal control animals. The smaller M / L ratio appeared to reflect depression of energy metabolism in the diabetic animals. Intrinsically lower and further decreased MDH activities may be factors that induce insulin resistance observed in diabetic cats.     

Decrease in malate dehydrogenase activities in peripheral leucocytes of type 1 diabetic dogs

The activities of the enzymes involved in the malate-aspartate shuttle were measured in peripheral leucocytes of dogs with type 1 diabetes mellitus. In the diabetic dogs, fasting plasma glucose concentrations were twofold greater than control levels despite insulin injections and the activities of malate dehydrogenase (MDH), which plays a crucial role in the malate-aspartate shuttle, were decreased remarkably. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity (M/L ratio) in leucocytes of the diabetic dogs was significantly lower than that of normal control dogs. The decrease of the M/L ratio appeared to reflect depression of energy metabolism in leucocytes of the diabetic dogs. The M/L ratio may be a useful parameter to evaluate metabolic conditions in diabetic dogs.     

Long term intensive exercise training leads to a higher plasma malate/lactate dehydrogenase (M/L) ratio and increased level of lipid mobilization in horses

Continuous high intensity training may induce alterations to enzyme activities related to glucose and lipid metabolism in horses. In our study, five Thoroughbred race horses (3 male and 2 female, avg age=5 yrs old) were compared against five riding horses (1 male, 1 female, 3 gelding, avg age=13 yrs old) in terms of energy metabolism, by examining plasma malate (MDH) and lactate (LDH) dehydrogenase activities and M/L ratio. MDH is involved in NADH and ATP generation, whereas LDH can convert NADH back into NAD(+) for ATP generation. An increase in plasma M/L ratio can reflect heightened energy metabolism in the liver and skeletal muscle of horses adapted to continuous intensive exercise. Moreover, plasma lipid metabolism analytes (adiponectin, NEFA, total cholesterol (T-Cho), and triglycerides (TG)) can reflect changes to lipolysis rate, which can also indicate a change in energy metabolism. Overall, race horses demonstrated increased MDH and LDH activity in plasma (4x and 2x greater, respectively), in addition to a plasma M/L ratio twice as high as that of riding horses (2.0 vs 1.0). In addition, race horses also demonstrated significantly higher levels of plasma NEFA (50% greater), TG (2x greater), and T-Cho (20% greater) as compared to riding horses. Therefore, race horse muscles may have adapted to prolonged high intensity endurance exercise by gaining a higher oxidative capacity and an increased capacity for fat utilization as an energy source, resulting in heightened energy metabolism and increased rate of lipid mobilization.     

Changes in hepatic enzyme activities in transgenic mice carrying human prototype c-Ha-ras gene treated with diethylnitrosamine

Activities of malate dehydrogenase (MDH) and aspartate aminotransferase in the malate-aspartate shuttle were significantly increased in the cytosolic fractions of livers with early neoplastic symptoms such as swelling and discoloration in transgenic mice carrying the prototype human c-Ha-ras gene, Tg-rasH2 mice, which were administered with diethylnitrosamine (DEN) as a carcinogenic chemical at a dose of 200 mg/kg body weight. Cytosolic MDH/LDH (lactate dehydrogenase) (ML) ratio increased significantly and was considered to be a useful marker to characterize the energy metabolism at early neoplastic stage in livers of the Tg-rasH2 mice.     

Changes in activities of enzymes related to malate-aspartate shuttle in leukocytes from dogs given a herb supplement.

Changes in activities of enzymes related to the malate-aspartate shuttle were measured in leukocytes (WBC) from dogs given food supplemented with 2 mg/kg of herb powder, Echevaria glauca, every day for 4 weeks. There were no significant differences in plasma concentrations of glucose, immunoreactive insulin, free fatty acids and triglyceride between dogs given food with or without the herb supplementation. Activities of malate dehydrogenase (MDH) and aspartate aminotransferase (AST) in the malate-aspartate shuttle increased remarkably in mitochondria of WBC from dogs fed the herb supplementation. It is suggested that Echevaria glauca herb supplementation might activate NADH shuttle systems and mitochondrial energy metabolism in dogs.     

Changes in Activities of Enzymes Related to Energy Metabolism in Peripheral Leukocytes of Fattening Steers

Glucose, triglyceride, cholesterol and immunoreactive insulin (IRI) concentrations, some enzyme activities in plasma, and activities of enzymes related to energy metabolism in peripheral leukocytes were measured in fattening Japanese Black Wagyu x Holstein steers fed on different diets at 8, 12, 16, 20 and 24 months of age. The plasma IRI concentrations at 20 and 24 months of age were significantly higher than those at 8 months of age. Activities of hexokinase (HK), glucose-6-phosphate dehydrogenase (G6PD), aspartate aminotransferase (AST), and malate dehydrogenase (MDH) in cytosolic fractions, and glutamate dehydrogenase (GLDH), MDH and AST in mitochondrial fractions in peripheral leukocytes of steers at 24 months of age were significantly higher than those at 8 months. Increasing plasma insulin concentration was considered to induce acceleration of glucose utilization in leukocytes of fattening steers. The cytosolic ratio of MDH/lactate dehydrogenase (LDH) activity in leukocytes increased significantly in the fattening process and was considered to be a useful indicator for evaluating changes in energy metabolism in steers.     

Thymoquinone enhances the activities of enzymes related to energy metabolism in peripheral leukocytes of diabetic rats

The aim of this study was to examine the effect of glycemic control using thymoquinone (TQ) on energy metabolism related enzymes in leukocytes of streptozotocin (STZ)-induced diabetic rats. The treatment of both TQ and insulin commenced 4 weeks after induction of diabetes. Plasma glucose, cholesterol and triglycerides levels were significantly reduced after TQ treatment, whereas immunoreactive insulin (IRI) showed significant increase. The activities of malate dehydrogenase (MDH) in cytosolic and mitochondrial fractions of peripheral blood leukocytes were significantly higher in rats treated with TQ and insulin as compared to that in diabetic controls. On the other hand the activities of lactic dehydrogenase (LDH) showed no significant changes between groups. ML ratio (cytosolic MDH/LDH specific activity ratio) was restored to those in the control rats. The results of this study demonstrate that TQ significantly increased insulin level and the activities of cytosolic and mitochondrial MDH in leukocytes of STZ-diabetic rats.     

Activities of Enzymes in the Malate–Aspartate Shuttle in the Peripheral Leukocytes of Dogs and Cats

The activities of the enzymes involved in the malate–aspartate shuttle and the expression of malate dehydrogenase (MDH), a rate-limiting enzyme in the NADH shuttle that produces ATP in glucose metabolism in leukocytes, were determined to investigate the differences in this shuttle system in the peripheral leukocytes of dogs and cats. There were no significant differences between dogs and cats in plasma glucose, immunoreactive insulin, free fatty acid or triglyceride concentrations. The activities of cytosolic and mitochondrial MDH and of mitochondrial glutamate dehydrogenase (GLDH) in canine leukocytes were significantly higher than in feline leukocytes. High activities of MDH in canine leukocytes were confirmed by RT-PCR analysis on the total RNA extracted from leukocytes. It was concluded that there were significant differences between dogs and cats in the NADH shuttle system.     

Changes in activities of enzymes related to energy metabolism in peripheral leukocytes of diabetic dogs with glycemic control by intensive insulin treatment

Changes in plasma glucose, immunoreactive insulin (IRI), and free fatty acids (FFA) concentrations and activities of enzymes related to energy metabolism in peripheral leukocytes were measured in type 1 diabetic dogs induced by streptozotocin administration before and after intensive insulin treatments. Before the insulin treatment, the plasma glucose concentrations were over 16.7 mmol/L and the activities of enzymes remarkably decreased in leukocytes of the diabetic dogs. In particular, malate dehydrogenase (MDH) activities decreased significantly. After glycemic control was achieved by insulin treatment, the plasma glucose concentrations were maintained between 6.8 and 7.7 mmol/L and the activities of enzymes restored to those in the control dogs. The changes in ML ratio (cytosolic MDH/lactate dehydrogenase specific activity ratio) reflected faithfully to metabolic conditions of animals and were considered to be a useful parameter to evaluate glycemic control and the effect of injected insulin in the diabetic dogs.     

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.     

LDH and CK isoenzyme patterns in the blood plasma of horses with elevated CK, LDH and AST activities

The distribution of LDH and CK isoenzymes in blood plasma of ten clinically sound Thoroughbreds with reasonable performance and without elevated clinico-chemical blood variables (reference group) was compared with 57 Thoroughbreds, which had histories of mild locomotor disturbances and/or poor performance and had elevated CK, LDH and/or AST activities (trial group). The trial group was subdivided according to the number of altered blood variables and in the groups with two as well as three altered blood variables also according to the extent of alteration of the total CK activity. The pattern of LDH and CK isoenzyme distribution in the blood plasma of the reference group was the following: 22% LDH1, 36% LDH2, 34% LDH3, 6% LDH4 and 2% LDH5 as well as 75% CK1 and 15% CK2. The remaining 10% of the plasma electropherogram could not be alloted to any one of the two CK bands. All trial groups built showed a similar pattern of changes in their isoenzyme distribution independent on kind and combination of altered enzyme activities. The shares of CK1, LDH4 and LDH5 were significantly increased whilst the shares of CK2, LDH1 and LDH2 decreased. A multiple analysis of variance demonstrated that only increased total CK activities had a pronounced effect on distribution of LDH and CK isoenzyme patterns in the trial group (p less than 0.01 for LDH2, LDH3, LDH4, CK1 and p less than 0.05 for CK2). The conclusion of the study was that the altered distribution pattern of LDH and CK isoenzymes of the trial group signalized an increased skeletal muscle membrane leakage.     

The Soft-tissue Components of the Vomeronasal Organ in Pigs, Cows and Horses

The soft-tissue components of the vomeronasal organ of the pig, the cow and the horse were studied with the aid of dissection, microdissection, and light microscopy and immunohistochemistry of series of transverse sections. In horses, the rostral end of the incisive duct was blind: thus, unlike in pigs and cows, there was no communication between the vomeronasal organ and the oral cavity. In all three species, the central part of the vomeronasal duct bore the ‘typical’ respiratory/ receptor epithelium lining on its lateral and medial walls. The rostral part of the duct was characterized by stratified columnar epithelium, while more caudal parts bore simple columnar type. The patterns of distribution of glands, blood vessels and nerves were closely associated with the patterns of distribution of duct linings. The distribution of soft-tissue components in pigs was less clearly defined than in cows and horses. Of the three species, nerves were detected in the rostral half of the vomeronasal parenchyma only in the horse.     

Changes in metabolite, energy metabolism related enzyme activities and peripheral blood mononuclear cell (PBMC) Populations in beef heifers with two differing liveweight change profiles in New Zealand

Metabolite and immunoreactive insulin (IRI) concentrations, energy metabolism related enzymes activities and peripheral blood mononuclear cell (PBMC) populations were measured in blood of pregnant Angus heifers with differing liveweight change profiles (gaining or losing), in New Zealand to investigate the meanings of those parameters in the restricted feeding beef heifers. Beef heifers losing liveweight (−412 g/day) showed significantly lower concentrations of plasma IRI, and higher concentrations of plasma free fatty acid (FFA) than heifers gaining liveweight (483 g/day). The cytosolic and mitochondrial malate dehydrogenase (MDH) activities and MDH/lactate dehydrogenase (M/L) ratio in leukocytes of the liveweight losing heifers were significantly higher than those the liveweight gaining heifers. Percentages of cluster of differentiation (CD) 3 positive cells and natural killer (NK) cells in PBMC decreased significantly in the liveweight losing heifers compared to those in the liveweight gaining heifers. Plasma IRI and FFA concentrations, leukocyte cytosolic and mitochondrial MDH activities and CD3 positive and NK cell populations may be useful markers to evaluate metabolic conditions and immunity in the restricted feeding beef heifers.