Sources of normal variation of plasma lactate dehydrogenase in range cattle.

1. Plasma levels of lactate dehydrogenase (LD) were estimated in Hereford cattle of 1 month to 12 yr of age kept under range conditions. 2. Plasma levels of LD were estimated in a group of confined cows and their calves fed a constant diet and kept in individual pens in the same geographic area as the range Herefords. 3. Large seasonal changes in mean plasma LD were observed in the range cattle but not in the confined, constant diet group. The seasonal changes corresponded with the appearance of new growth on the range. 4. Changes in plasma LD were observed at weaning time in the range cattle but not in the confined group. 5. Sex had no effect upon plasma LD levels in the juvenile animals of either group. No comparison of adult animals could be made. 6. Mean plasma LD levels did not change with age but the seasonal effect decreased in magnitude as the animals matured. 7. An increase in plasma LD was observed around the first ovulation in the confined animals. 8. A decrease in plasma LD was observed during parturition in the confined group.     

Cell cycle related studies on thymidine kinase and its isoenzymes in Ehrlich ascites tumours

Thymidine kinase (TK) activity was measured in relation to the cell cycle of in vivo growing ascites tumour cells. The cells were synchronized by means of centrifugal elutriation and the cell cycle composition of the cell fractions was determined by flow cytometry. TK activity was low in G1, increased during S phase and declined in G2. A half-life of TK activity of about 45 min was found throughout the cell cycle. Four isoenzymes at pI values of 4.1, 5.3, 6.9 and 8.3, denoted as isoenzymes 1-4, were identified using isoelectric focusing. Isoenzymes 3 and 4 were responsible for the profound cell cycle related changes in the TK activity. Corresponding isoenzymes were also found in the fetal mouse liver. In the adult mouse liver isoenzyme 2 was the dominating isoenzyme. The half-life of the isoenzymes was in the same range as for the total TK activity. We conclude that the low TK activity in G1 is due to degradation of the enzyme in G2 at a normal rate combined with an arrest in the synthesis of TK. We also conclude that isoenzyme 4 and the intermediate isoenzyme 3, which had earlier been suggested to be a mitochondrial form of TK, in fact represent cytoplasmatic forms of TK. According to cell cycle and pI studies, isoenzyme 2 belongs to the mitochondrial form. Studies with various phosphor donors and specific substrates, however, indicate that it also contains a cytoplasmic component.     

Frequent moving of grazing dairy cows to new paddocks increases the variability of milk fatty acid composition

The aim of this work was to investigate the variations of milk fatty acid (FA) composition because of changing paddocks in two different rotational grazing systems. A total of nine Holstein and nine Montbéliarde cows were divided into two equivalent groups according to milk yield, fat and protein contents and calving date, and were allocated to the following two grazing systems: a long duration (LD; 17 days) of paddock utilisation on a heterogeneous pasture and a medium duration (MD) of paddock utilisation (7 to 10 days) on a more intensively managed pasture. The MD cows were supplemented with 4 kg of concentrate/cow per day. Grazing selection was characterised through direct observations and simulated bites, collected at the beginning and at the end of the utilisation of two subsequent MD paddocks, and at the same dates for the LD system. Individual milks were sampled the first 3 days and the last 2 days of grazing on each MD paddock, and simultaneously also for the LD system. Changes in milk FA composition at the beginning of each paddock utilisation were highly affected by the herbage characteristics. Abrupt changes in MD milk FA composition were observed 1 day after the cows were moved to a new paddock. The MD cows grazed by layers from the bottom layers of the previous paddock to the top layers of the subsequent new paddock, resulting in bites with high organic matter digestibility (OMD) value and CP content and a low fibre content at the beginning of each paddock utilisation. These changes could induce significant day-to-day variations of the milk FA composition. The milk fat proportions of 16:0, saturated FA and branched-chain FA decreased, whereas proportions of de novo-synthesised FA, 18:0, c9-18:1 and 18:2n-6 increased at paddock change. During LD plot utilisation, the heterogeneity of the vegetation allowed the cows to select vegetative patches with higher proportion of leaves, CP content, OMD value and the lowest fibre content. These small changes in CP, NDF and ADF contents of LD herbage and in OMD values, from the beginning to the end of the experiment, could minimally modify the ruminal ecosystem, production of precursors of de novo-synthesised FA and ruminal biohydrogenation, and could induce only small day-to-day variations in the milk FA composition.     

Lactate dehydrogenase isoenzyme patterns as a method of pregnancy detection in cattle

The objective of this study was to evaluate serum lactate dehydrogenase isoenzyme patterns in open and pregnant Holstein and Hereford cows as a method of detecting pregnancy. Serum samples were collected from 26 Holstein and 13 Hereford cows and lactate dehydrogenase isoenzyme patterns were examined by electrophoresis and quantitated by scanning densitometry. Lactate dehydrogenase isoenzyme(4) and LDH(5) were found in higher concentration (P 相似文献   

Polyacrylamide gel electrophoresis of rat brain acetylcholinesterase: isoenzymes of normal rat brain

Abstract— Triton-solubilized acetylcholinesterase (EC 3.1.1.7) of rat brain was submitted to vertical flatbed polyacrylamide gel electrophoresis. Three anodally migrating isoenzyme zones with low relative mobilities could be resolved, each of which on quantitative densitometry appeared to consist of more than one subzone. More than 50 per cent of the total AChE activity was exhibited by the isoenzyme zone closest to the origin (isoenzyme zone 3). Regional differences in AChE isoenzyme activity were quantitative only with the caudate-putamen complex, midbrain, pons and medulla oblongata exhibiting relatively high content of the three isoenzymes and the cerebral cortex and olfactory bulb possessing weak isoenzyme activities. Intermediate levels of isoenzyme activities were observed in the cerebellum and hippocampus. In all areas examined, the relative percentage values for each isoenzyme remained constant. AChE isoenzymes from the forebrain, brain stem and cerebellum of 15- and 30-day-old rats appeared to have identical patterns. In brain stem, no quantitative differences could be detected in the isoenzyme activities between 15 and 30 days of age. At both ages, the isoenzymes of male and female rats did not show any qualitative differences. The single cholinesterase (EC 3.1.1.8) isoenzyme which could be identified in brain stem supernatants of 30-day-old rats was weakly reactive and appeared to have the same relative mobility as the major acetylcholinesterase zone, zone 3. Acetylcholinesterase isoenzymes failed to demonstrate any differential response toward varying concentrations of inhibitors and to changes in pH. While there were basic similarities in the acetylcholinesterase and cholinesterase isoenzyme patterns of brain, serum, liver, skeletal muscle and intestine, brain alone exhibited a marked preponderance of the acetylcholinesterase isoenzyme zone 3.     

Comparison of biochemical markers between benign and malignant ovarian cysts

To investigate the presumption that earlier diagnosis of ovarian tumors might lead to an improved outcome, we compared several substances in the fluid of benign and malignant ovarian cysts. Our results showed the following: (1) benign cysts were readily separated from malignant cysts on the basis of total lactate dehydrogenase (LD) activity and except for mucinous cysts, by their isoenzyme patterns. (2) Preoperative serum total LD activity and LD isoenzymes were of no diagnostic help in predicting the malignant tumors. (3) Carcinoembryonic antigen (CEA) levels usually differed greatly between benign and malignant cysts, although the benign mucinous cysts had CEA levels indistinguishable from malignant cysts. (4) Prominent quantitative differences between Roche and Abbott CEA activity were present in both benign and malignant cysts. (5) Preoperative serum CEA levels were not helpful in differentiating the benign from the malignant cysts.     

Organ specificity and lactate-dehydrogenase activity. Some properties of human spermatozoal lactate dehydrogenase

1. The presence of a characteristic lactate-dehydrogenase isoenzyme (LD(x)) in human, mouse and dog testis and in human spermatozoa has been confirmed by electrophoresis on cellulose acetate and on polyacrylamide gel. 2. The human spermatozoal isoenzyme exhibits a much higher affinity for 2-oxobutyrate than any of the five isoenzymes found in other tissues. K(m) values of 0.05mm for pyruvate and 0.18mm for 2-oxobutyrate were obtained. 3. LD(x) differs from other lactate-dehydrogenase isoenzymes in that its properties cannot be correlated with its electrophoretic mobility. It resembles LD(1) in being strongly inhibited by 0.2mm-oxalate and relatively resistant to 2m-urea, and in being relatively stable to heat. 4. The surprisingly high activity of LD(x) with 2-oxobutyrate suggests that this substance or 2-hydroxybutyrate may play a part in spermatozoal metabolism.     

Carbonic anhydrase isoenzymes in the erthrocytes and dorsolateral prostate of the rat

1. Three forms of the zinc-containing enzyme carbonic anhydrase (EC 4.2.1.1) were isolated from the erythrocytes of the rat and two forms from the dorsolateral prostate of the rat. Several additional minor components were observed but not isolated. Separation of the isoenzymes was achieved by ion-exchange chromatography, polyacrylamide-gel electrophoresis and isoelectric focusing. 2. The general properties of the isolated isoenzymes, their molecular weights and their contents of zinc were closely similar. As catalysts of the hydration of carbon dioxide, however, they were distinctly different. The two most abundant isoenzymes of the erythrocytes, which were found in equal proportions, differed 70-fold in specific activity, whereas the isoenzymes of the dorsolateral prostate were similar to one another and resembled the high-activity component of the erythrocytes. The inhibition of the latter by acetazolamide (5-acetamido-1-thia-3,4-diazole-2-sulphonamide) was mainly competitive, whereas in identical conditions the low-activity erythrocyte component and the dorsolateral prostate isoenzymes were non-competitively inhibited. 3. The use of chloroform-ethanol to remove haemoglobin from the rat haemolysate was found (a) to bring about changes in the kinetic properties of the soluble isoenzymes and (b) to cause the appearance of an additional isoenzyme. 4. The actions were compared of the inhibitors acetazolamide, 1,1-dimethylaminonaphthalene-5-sulphonamide and ethoxzolamide (6-ethoxybenzothiazole-2-sulphonamide) on the hydrolysis of p-nitrophenyl acetate catalysed by the isoenzymes. 5. The low-activity erythrocyte isoenzyme was an efficient catalyst of the hydrolysis of beta-naphthyl acetate whereas the high-activity forms were much less active towards this ester. Neither of the isoenzymes present in the dorsolateral prostate catalysed this reaction. 6. Carbonic anhydrase in the rat dorsolateral prostate accounts for no more than 5% of the unusually high content of zinc in this organ.     

谷子耐盐系的几种同工酶的变化及外源ABA对它们的影响

对谷子细胞变异系的过氧化物酶,细胞色素氧化酶和脂酶同工酶的分析表明：耐盐系在无盐胁迫条件下过氧化物酶同工酶的总酶活性高于对照系,其各同工酶带的强度也与对照有明显差异;在ＮａＣｌ迫条件焉耐盐系中个别原有的酶带消失。细胞色素氧化活性与过氧化物酶同工酶的活性变化情况类似,均与对照有明显差异。     

Changes in the activity of adenylate kinase isoenzymes in the ischemic rabbit heart

Under the influence of 1 hour myocardial ischemia activity of rabbit heart mitochondrial isoenzyme AK2 increased by 40%, but the activity of matrix isoenzyme AK3 decreased by 77%. No changes were found both in total adenylate kinase activity, and cytosolic isoenzyme AK1. The reasons of these alterations are not sufficiently clear. Apparently, they are related with functioning conditions of these isoenzymes in ischemic tissue.     

3-Deoxy-D-arabino-heptulosonic acid 7-phosphate synthase mutants of Salmonella typhimurium.

The first committed step of aromatic amino acid biosynthesis in Salmonella typhimurium was shown to be catalyzed by three isoenzymes of 3-deoxy-D-arabino-heptulosonic acid 7-phosphate (DAHP) synthase. Mutations in each of the genes specifying the isoenzymes were isolated and mapped. aroG, the structural gene for the phenylalanine-inhibitable isoenzyme, was linked to gal, and aroH, the structural gene for the tryptophan-inhibitable isoenzyme, was linked to aroE. aroF, the structural gene for the tyrosine-inhibitable isoenzyme, was linked to pheA and tyrA, which specify the phenylalanine- and tyrosine-specific branch-point enzymes, respectively. The phenylalanine-inhibitable isoenzyme was the predominant DAHP synthase in wild-type cells, and only the tryosine-inhibitable isoenzyme was completely repressed, as well as inhibited, by low levels of its allosteric effector. The DAHP synthase isoenzymes were separated by chromatography on diethylaminoethyl-cellulose with a phosphate gradient which contained enolpyruvate phosphate to protect the otherwise unstable phenylalanine-inhibitable isoenzyme. No cross-inhibition of either the tyrosine- or phenylalanine-inhibitable isoenzyme was observed at inhibitor concentrations up to 1 mM. The tryptophan-inhibitable isoenzyme was partially purified from extracts of a strain lacking the other two isoenzymes and shown to be inhibited about 30% by 1 mM tryptophan. A preliminary study of interference by tryptophan in the periodate-thiobarbiturate assay for DAHP suggested a combined effect of tryptophan and erythrose 4-phosphate, or an aldehydic compound resulting from degradation of erythrose 4-phosphate by periodate.     

Sources of normal variation of plasma l-aspartate:2-oxoglutarate aminotransferase in Hereford range cattle

1. Plasma levels of L-aspartate:2-oxoglutarate aminotransferase (AST) were estimated in Hereford cattle, 1 month to 12 years of age, kept under range conditions and in a group of Hereford x Angus cows kept on the same range. 2. Plasma levels of AST were estimated in a group of Crossbred cows and their calves fed a constant diet and kept in individual pens in the same geographic area as the Herefords. 3. Seasonal changes in mean plasma AST were observed in the Herefords corresponding with the change from dry winter grasses/hay and well water to fresh spring and summer grasses and slough water. No seasonal changes were observed in the Crossbreds given a constant dry diet and city water. 4. Plasma AST increased with age in calves 1 to 12 months of age in the Herefords but not in the Crossbreds. Mean plasma AST did not change with age in any of the adult cattle studied. 5. Small increases in plasma AST corresponding to increases in ambient temperature above - 12 degrees C were observed in the Crossbreds. 6. An increase in plasma AST was observed near the time of first ovulation in the confined cows. 7. No relationship could be demonstrated between plasma AST and sex, breed or time to parturition in the range cattle. Breed differences were observed in the Crossbred cows.     

Development of (1-->3,1-->4)-beta-d-Glucan Endohydrolase Isoenzymes in Isolated Scutella and Aleurone Layers of Barley (Hordeum vulgare)

An immunological assay has been used to investigate the synthesis of (1→3,1→4)-β-glucanase (EC 3.2.1.73) isoenzymes from isolated barley aleurone layers and scutella. Enzyme release from both tissues is enhanced by 1 micromolar gibberellic acid and 10 millimolar Ca²⁺, although increases induced by gibberellic acid are observed only in the presence of Ca²⁺. Isoenzyme I is synthesized predominantly in the scutellum, while isoenzyme II is synthesized exclusively in the aleurone. A third, putative isoenzyme III has been detected in significant proportions in scutellar secretions and may also be secreted from aleurone layers. Both gibberellic acid and Ca²⁺ appear to preferentially enhance isoenzyme II secretion from the aleurone and isoenzyme III secretion from scutella. The patterns of isoenzyme secretion are suggestive of tissue-specific differences in expression of the genes which code for (1→3,1→4)-β-glucanase isoenzymes. Qualitatively similar results were obtained with barley cultivars harvested in Australia and North America.     

Developmental changes in creatine phosphokinase isoenzymes in neonatal mouse hearts.

The distribution of creatine phosphokinase isoenzymes differs in extracts of newborn and adult mouse hearts. Electrophoresis on acetate strips reveals the presence of BB, MB, and MM isoenzymes in the 2 day old neonate heart, with relative activities of 4%, 24% and 72% respectively. Beginning at 6 days of age, a fourth isoenzyme, shown to be associated with mitochondria, is seen moving toward the cathode. With age the distribution changes, with BB disappearing by 18 days. By 25 days the relative proportions of MB, MM and mitochondrial CPK have reached 5%, 86% and 9%, respectively, similar to the levels seen in the adult. The late appearance of the mitochondrial isoenzyme may reflect a difference in the requirement of the developing and adult heart for ATP and phosphocreatine.     

Purification and partial characterization of high and low activity carbonic anhydrase isoenzymes from Malaclemys terrapin centrata.

1. High activity (CA C) and low activity (CA B) carbonic anhydrase isoenzymes have been purified from turtle erythrocytes. 2. The two isoenzymes differed in CO2 hydration specific activity by 36-fold. 3. The low activity isoenzyme contained one half-cystine residue, whereas the high activity isoenzyme contained four half-cystines and required a reducing environment to maintain activity. Both isoenzymes contained zinc. 4. Molecular weights of 28,500 and 30,400 daltons were established for the low and high activity isoenzymes respectively. 5. Both isoenzymes were inhibited by acetazolamide, but only the high activity isoenzyme was inhibited by parachloromercuribenzoate. 6. The low activity isoenzyme was present in the erythrocytes at about 8-10 times the concentration of the high activity isoenzyme. 7. The high activity isoenzyme cross-reacted with antibodies prepared against pure chicken carbonic anhydrase C.     

Hexokinase isoenzymes in tissues of the adult and developing guinea pig

Changes in the activities and isoenzyme distribution of hexokinase were determined in a number of tissues during the development of the guinea pig. The total activity in the fetal liver showed a large fall during the second half of gestation to reach adult values by term. With normal diet the fetal, neonatal, and adult livers had isoenzymes I and III but little or no detectable IV (glucokinase). The fetal liver had predominantly type I, but the proportion of type III increased during development. The kinetics of the guinea pig isoenzymes were similar to those reported for the rat. Two additional isoenzymes with mobility between I and II were detected in the fetal liver and blood. They appear to have kinetic properties similar to type I. Detectable liver glucokinase activity was induced by glucose administration to adult guinea pigs. The total activity in kidney, brain and skeletal muscle showed a postnatal rise while in the fetal heart it was high and declined after birth. These tissues contained predominantly type I with varying proportions of type III hexokinase. The ratio of particulate-bound to soluble hexokinase varied from tissue to tissue. All except the liver showed a significant increase in binding after birth. The changes are discussed in relation to the control of glucose utilization in the fetal and neonatal periods.     

Variability of antioxidant enzyme activity and isoenzyme profile in needles of Serbian spruce (Picea omorika (Panč.) Purkinye)

Variations were studied of the activity and isoenzyme patterns of soluble peroxidase, catalase, catechol oxidase and superoxide dismutase, in needles of the Balkan endemic conifer Serbian spruce, Picea omorika (Pan?.) Purkinye. The samples were collected from the natural habitat of the species, Mt. Tara. Seasonal changes were found to affect enzymatic activities and isoenzyme profiles. Total protein content was significantly lower in the summer than in other seasons. Several isoforms of peroxidase, catechol oxidase and superoxide dismutase (SOD), as well as two catalase isoenzymes were detected. The number of peroxidase isoenzymes was greatest during the vegetative season. Catalase and catechol oxidase peaked in summer and spring, respectively. Total SOD and Mn-SOD activities were significantly higher in the winter samples than the summer ones.     

The regulation of mitochondrial-bound hexokinases in the liver

A functional coupling between bound hexokinase and the inner mitochondrial compartment has been shown. It is based structurally on the binding of hexokinase to a pore protein which is present in zones of contact between the two boundary membranes. The latter was observed by electron microscopic localization of antiporin and hexokinase at the mitochondrial surface. The four isoenzymes present in liver differ considerably in their activity after binding to the mitochondrial surface. This was found by binding studies using the four isoenzymes isolated from the supernatant. Isoenzyme IV did not bind at all. Isoenzymes I-III did bind and became activated: I, 5.9-fold; II, 39-fold; and III, 1.3-fold. These results suggest that the in vivo activity of hexokinase in the mitochondrial fraction is much larger than so far observed. Furthermore the binding of isoenzymes was differently affected by metabolites. Glucose-6-phosphate exclusively desorbed isoenzyme I from the mitochondrial membrane whereas free fatty acids predominantly liberated isoenzymes II and III. A reciprocal change of the levels of free fatty acids and glucose 6-phosphate in livers of starved rats therefore, can explain why exclusively mitochondrial-bound isoenzymes II and III decreased 10-fold while at the same time isoenzyme I increased.     

Creatine phosphokinase isoenzymes and Trypanosoma cruzi infections.

1. Substantial increases in total creatine phosphokinase (CPK) and in isoenzymes from heart (CPK-MB) and skeletal muscle (CPK-MM) were observed during acute infections with the House 510 and House 11 strains of Trypanosoma cruzi. 2. In infections with the reticulotropic Tulahuen strain total CPK levels were lower and the isoenzyme pattern was essentially normal. 3. Gamma-glutamyl transpeptidase was considerably increased in the Tulahuen but not in the House 510 and House 11 infections. 4. These findings are useful in assessing tissue damage during T. cruzi infections and they also demonstrate differences between myotropic and reticulotropic strains which may aid in their taxonomic classification.