Isoenzymes of Lactate Dehydrogenase in Swine Stability During Storage at Different Temperatures and By Heat Treatment

The isoenzymes of lactate dehydrogenase (LDH) in serum from normal pigs were studied after separation by agar gel electrophoresis with subsequent staining with a tetrazolium salt. Experiment 1. The stability of isoenzymes was investigated for 5 successive days after storage at room temperature (22°C), in the refrigerator (4°G), and once after storage for 32 days in the deep-freezer (—20°C). Greatest loss of activity was seen after storage in the refrigerator, where LDH2 and LDH3 lost most of its activity after 5 days. In LDH4 and LDH5 no loss had occurred at this time. Also at room temperature great losses were seen in LDH2 and LDH3. After storage in the deep-freezer an increase in LDH3 activity was recorded. Experiment 2. Serum samples were kept in water baths for 30 min. at 50, 53, and 56°C. A simultaneous and increasing loss in activity of LDH3, LDH4, and LDH5 was seen from 50° to 56°C. At 56° no activity was left in LDH3, LDH4, or LDH5, and only about 15 % of the original activity was present in LDH2. LDH1 showed no loss at 56°, but all activity was lost at 65°. A close correlation was found between total lactate dehydrogenase and α-hydroxybutyrate dehydrogenase activity in both experiments.     

鱼类乳酸脱氢酶同工酶聚丙烯酰胺凝胶电泳技术研究

本文对用聚丙烯酰胺凝胶电泳（ＰＡＧＥ）分离鱼类乳酸脱氢酶（ＬＤＨ）同工酶的技术进行了研究。结果表明：该方法优于淀粉凝胶电泳及琼脂糖凝胶电泳分析鱼类ＬＤＨ同工酶,具有较高的分辩率。     

Stability of Some Serum Enzymes in Sheep,Cattle, and Swine During Storage at Different Temperatures

Owing to the increased use of serum enzyme determinations in veterinary diagnostic work, greater knowledge about the keeping qualities of different animal sera under various storing conditions seems desirable. The present paper deals with the stability of serum aspartate aminotransferase (AspAT = GOT), alanine aminotransferase (A1AT = GPT), lactate dehydrogenase (LDH), and a-hydroxybutyrate dehydrogenase (HBD) in cattle, sheep, and swine. Sera from 14—16 animals of each species were analysed daily for 5 days after storage at room temperature (22°C) and in the refrigerator (4°C). Samples kept in the deep-freezer (—20°C) were reanalysed once after 32—38 days. Significant differences of serum activity were found between individuals for all enzymes in the three species. Great variations were found in the stability of enzyme activities of different species. To summarize, it may be said that the changes of transferase activities were less pronounced under the different storing conditions than those of the dehydrogenases investigated. Pig serum in particular showed heavy losses of the latter enzymes already after 1 day, more pronounced at refrigerator than at room temperature. As a consequence of the results obtained, practical recommendations for analytical work on these enzymes are suggested.     

Diurnal and Seasonal Variations of Serum Enzyme Activity in Cattle and Sheep

In order to test the variation of enzyme activity in serum of cattle and sheep during the day, blood samples were taken at three hrs. interval from 6 a.m. to 9 p.m. The following enzymes were assayed: Aspartate aminotransferase (AspAT = GOT), alanine aminotransferase (AlAT = GPT), total lactate dehydrogenase (LDH), and a-hydroxybutyrate dehydrogenase (HBD). The variation between animals contributed by far to the greatest part of the total variation in clinical healthy animals. The time-of-day-dependant variation was less than 3 %, except for alanine aminotransferase. During the first two weeks of spring pasture serum aspartate and alanine aminotransferase levels were significantly raised in both cows and ewes, compared with serum levels of the same animals on indoor feeding. No such increase occurred in total lactate dehydrogenase.     

牛血Cu/Zn-SOD的热稳定性

研究了不同的Cu^2 和Zn^2 浓度配比对SOD热稳定性的影响,并测定了SOD活性及利用考马斯亮蓝D250法测定蛋白质含量。结果表明在Cu^2 、Zn^2 分别为5．4mmol／1．和3．6mmol／L时SOD的活性最高,达到了92．6％;在75℃时大部分杂蛋白变性沉淀,SOD的比活力最高。最后确定在加入5．4mmol／L．Cu^2 ,3．6mmol／LZn^2 条件下,75℃下热变性,可以取得比较高活性和比活性的SOD。从而为寻找更简便、经济的提纯方法提供参考依据。     

Evidence of Lactate Dehydrogenase in Anaplasma marginale

Extracts from preparations of partially purified Anaplasma marginale revealed low levels of lactate dehydrogenase (LDH). Enzyme inhibition by immune sera further indicated that A. marginale possesses a protein moiety the same as that of the normal red blood cell (RBC), although data suggested an alteration of LDH(1) from that observed in normal RBC. Bimodal isozyme distribution was detected after electrophoresis of the extracts. One isozyme approached the cathode and the other the anode, and both appeared to be nicotinamide adenine dinucleotide-dependent. Heterogeneity of parasite and host cell isozymes was established on the basis of zone electrophoresis on cellulose acetate strips.     

Activity of Lactate Dehydrogenase in Serum and Cerebral Cortex of Immature and Mature Rats after Hypobaric Hypoxia

In our previous studies we have found both an increase of lipid peroxidation damage (expressed as levels of thiobarbituric acid-reactive substances) in brain and plasma lactate concentration in 21-day-old rats after a 30-min exposure to hypobaric hypoxia. Pretreatment of rats with l-carnitine decreased both parameters. The aim of our present study was to determine if the l-carnitine-dependent decrease of plasma lactate could be due to a modification of lactate dehydrogenase (LDH) activity. We followed brain and blood serum LDH activity of 14-, 21- and 90-day-old Wistar rats. We found an increase of brain LDH activity with age. However, we did not observe any significant differences in LDH activity after exposure to hypobaric hypoxia or l-carnitine pretreatment. In contrast to brain, serum LDH activity did not show any clear age-dependence. The hypoxia exposure increased LDH activity of 21-day-old rats only. Pretreatment of rats with l-carnitine decreased serum LDH activity of 21- and 90-day-old rats probably due to membrane stabilizing role of l-carnitine. In conclusions, acute hypobaric hypoxia and/or l-carnitine pretreatment modified serum but not brain LDH activity.     

Malate Dehydrogenase Isoenzymes in Cotton Leaves of Different Ages

Malate dehydrogenase isoenzymes from the blades of different aged leaves of the cotton plant have been investigated. The total extractable malate dehydrogenase activity varied widely between leaves of different ages and different locations on the plant. Malate dehydrogenase zymograms developed from the extracts which contained significantly different levels of enzyme activity appear to indicate the presence of different groups of malate dehydrogenase isoenzymes in leaves of different ages. However, under appropriate conditions of polyacrylamide gel electrophoresis, the same number of malate dehydrogenase isoenzymes with the same relative mobilities were detected in all the leaves studied. These findings are discussed in relation to reports that malate dehydrogenase isoenzymes change with plant development or that they have different roles in the plant.     

Glucose-6-Phosphate Dehydrogenase Activity and Glutathione Stability in Fetal and Adult Bovine Erythrocytes

Previously, we found a substantially higher glucoses-phosphate dehydrogenase (G6PD) activity and a slightly higher 6-phosphogluconate dehydrogenase (6PGD) activity in bovine fetal erythrocytes than in bovine adult erythrocytes (Steensgaard 1968). Now, we have investigated whether these differences in dehydrogenase activities were followed by characteristic differences in glutathione (GSH) stability and glutathione concentration. The results are shown in Table 1, which also gives the results of the same investigations on normal and G6PD deficient human erythrocytes.     

藏羚羊与藏绵羊心肌、骨骼肌中肌红蛋白含量和乳酸脱氢酶活性的比较

为了探讨藏羚羊(Pantholops hodgsonii)对低氧环境的适应机制。以生活在同海拔(4 300 m)的藏绵羊(Tibetan Sheep)为对照,用分光光度法测定2种动物心肌、骨骼肌中肌红蛋白(myoglobin,Mb)含量、乳酸(lactic acid,LD)含量及乳酸脱氢酶(lactate dehydrogenase,LDH)活力。结果显示,藏羚羊心肌和骨骼肌中Mb含量明显高于藏绵羊(P0.05),但心肌和骨骼肌的Mb含量无差别(P0.05),而藏绵羊心肌Mb含量明显高于骨骼肌(P0.05);藏羚羊心肌和骨骼肌中LD含量及LDH活力明显低于藏绵羊(P0.05),且2种动物心肌中的LDH活力均明显低于其骨骼肌(P0.01)。结果表明,藏羚羊尽管生活在高寒缺氧地区,其心肌和骨骼肌细胞仍然能得到丰富的氧供应,并非处于缺氧状态,这可能是通过增加心肌和骨骼肌中Mb的含量,提高其在低氧环境获取和储存氧的能力,从而提高有氧获能水平。与之相反,藏绵羊尽管也生活在高寒缺氧地区,但其心肌和骨骼肌中Mb含量相对于藏羚羊较低,且LD含量和LDH活力较高,说明其心肌和骨骼肌细胞内氧供不如藏羚羊丰富,提示藏绵羊可能主要以糖酵解获能。我们推测这种差异可能与两种动物不同的运动习性密切相关,且认为藏羚羊较高的Mb含量可能是其适应高原缺氧条件的分子基础之一。     

精子特异性乳酸脱氢酶的免疫学特性及其应用

精子特异性乳酸脱氢酶特异地存在于鸟类和哺乳类动物的成熟睾丸和精子中,为精子的运动和存活提供能量,它是一种自身抗原,其天然抗体不与体细胞LDH同工酶发生交叉反应,用LDH－C4免疫小鼠或免疫等能够诱导免疫应答,导致生育率的降低,因此在人类避免和鼠害控制方面将有较好的应用前景。     

血清乳酸脱氢酶评估颅脑手术患者重症肺炎的临床价值

摘要 目的：分析血清乳酸脱氢酶在评估颅脑手术患者重症肺炎的中的应用价值。方法：选取2020年7月~2022年6月在本院神经外科进行收治的颅脑择期手术患者268例,根据是否合并术后肺炎,分为非感染组（n=185）和肺炎组（n=83）,肺炎组根据肺炎严重程度分为重症肺炎31例及普通肺炎52例。采用酶联免疫吸附法测定血清PCT、CRP水平;采用酶比色法检测血清 LDH 水平,采用Pearson相关性检验分析合并肺炎患者血清LDH、PCT、CRP水平与病情严重程度的相关性。采用ROC曲线分析血清LDH、PCT、CRP水平对颅脑手术患者合并重症肺炎的临床诊断价值,血清LDH、PCT、CRP水平预估颅脑手术合并重症肺炎患者预后的价值。结果：术后1 d、3 d、7 d与未感染组对比,普通肺炎组患者血清LDH、PCT、CRP水平均明显升高（P<0.05）;与普通肺炎组对比,重症肺炎组患者血清LDH、PCT、CRP水平均明显升高（P<0.05）。经Pearson相关性检验分析,颅脑手术合并肺炎患者血清LDH、PCT、CRP水平与PSI评分呈明显正相关（P<0.05）。以术后1 d血清LDH、PCT、CRP水平建立ROC曲线,结果发现,血清LDH、PCT、CRP的曲线下面积分别为0.840、0.825、0.746。以术后7 d血清LDH、PCT、CRP水平建立ROC曲线,结果发现,血清LDH、PCT、CRP的曲线下面积分别为0.819、0.725、0.684。结论：LDH在颅脑手术合并肺炎患者中高表达,与肺炎严重程度正相关。LDH在区分非重症肺炎患者和重症肺炎患者以及预测重症肺炎患者的28天死亡率方面具有良好价值。     

牛蛙乳酸脱氢酶同工酶的分离纯化及其性质研究

牛蛙心脏中乳酸脱氢酶在聚丙烯酰胺凝胶电泳上显示3种同工酶区带,分别命名为LDH1、LDH2、LDH3,其中LDH1的活力占绝对优势.采用HiTrap^TM Blue HP 亲和层析和DEAE-Sephadex A离子交换层析对牛蛙骨骼肌中的LDH3进行了分离纯化.纯化的LDH3比活力为295 U/mg,Km NADH=0.028,Km丙酮酸=1.242,在SDS-PAGE上显示两条带,提示该同工酶是由两种亚基组成的,亚基的分子量分别为35.3 kD和37.6 kD.     

Reference Values for Serum Magnesium Levels in Young Adult Iranian Subjects

This study aims at determining the reference values for serum magnesium (Mg) concentrations in Iranian adults. Serum Mg level was measured using flame atomic absorption spectrometry in 491 subjects (233 men and 258 women), aged 20-50 years, randomly selected from a population-based study. The International Federation of Clinical Chemistry guidelines and the robust method were used for determining the reference values. The 95% reference values for serum Mg concentration were 1.83-2.49, 1.79-2.48, and 1.83-2.55 mg/dL in men, women, and total population, respectively. The prevalences of hypo- and hypermagnesemia, according to the reference values obtained in the current study, were 2.5% and 4.0%, respectively. In conclusion, this study reports serum Mg reference values based on current standards in a large healthy population of young Iranian adults.     

Lactate Dehydrogenase Activity in Certain Strains of Staphylococcus aureus

Lactate dehydrogenase (LDH) was studied in phage-propagating strains 29, 3A, 6, 81, and 42D of Staphylococcus aureus selected from the five groups in the International-Blair series. Cells were cultivated in Brain Heart Infusion (Difco) under nearly anaerobic conditions and were harvested near the end of the log phase. LDH activity was maximal at the end of the exponential growth period and was measured spectrophotometrically by reduction of p-nitro-blue tetrazolium, with phenazine methosulfate as a coupling agent. Crude enzyme extracts were prepared both by an acetone extraction technique and by sonic treatment. LDH activity for these enzyme preparations was determined by the colorimetric method mentioned and also by measuring the rate of nicotinamide adenine dinucleotide reduction at 340 mmu. The order of activity observed, by use of both assay methods, was 29 > 81 > 6 > 3A > 42D. LDH forms (possibly isoenzymes) for each of 15 strains, which represent the five phage-propagating groups of the International-Blair series, were separated by acrylamide gel electrophoresis. Five forms were distinguished and arbitrarily numbered on the basis of their rate of migration, no. 5 being the slowest component. No one strain had more than four, nor fewer than two, LDH forms. Form 3 appeared in 13 of the 15 strains and was followed in frequency by no. 2, 1, 4, and 5.     

Coimmobilization of Lactate Dehydrogenase and Low-Molecular-Weight Thiols on Sepharose

Lactate dehydrogenase (EC 1.1.1.27) and dithiothreitol (DTT) were coimmobilized on Sepharose activated with cyanogen bromide. It was demonstrated that the addition of 10 mM DTT (but not 2-mercaptoethanol) during immobilization increased the enzyme specific activity 1.5–5-fold depending on the initial extent of Sepharose activation by cyanogen bromide. The total activity increased two- to threefold. The lactate dehydrogenase preparations were rich in matrix-immobilized sulfhydryl groups (1.8–13.0 nmol per ml gel). The presence of DTT increased the stability of immobilized lactate dehydrogenase.     

Properties of Lactate Dehydrogenase in a Psychrophilic Marine Bacterium

Lactate dehydrogenase (EC 1.1.1.27) from Vibrio marinus MP-1 was purified 15-fold and ammonium activated. The optimum pH for pyruvate reduction was 7.4. Maximum lactate dehydrogenase activity occurred at 10 to 15 degrees C, and none occurred at 40 degrees C. The crude-extract enzyme was stable between 15 and 20 degrees C and lost 50% of its activity after 60 min at 45 degrees C. The partially purified enzyme was stable between 8 and 15 degrees C and lost 50% of its activity after 60 min at 30 degrees C. The thermal stability of lactate dehydrogenase was increased by mercaptoethanol, with 50% remaining activity at 42 degrees C.