OBJECTIVETo explore the adaptive mechanism to hypoxia in skeletal muscle of tibetan antelope.

METHODSTibetan sheep which living at the same altitude (4 300 m) with tibetan antelope and low altitude (1 800 m) sheep as control, the content of myoglobin (Mb) and lactic acid (LA), the activity of lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) in skeletal muscles among three animals were analyzed by spectrophotometer.

RESULTSThe content of myoglobin in skeletal muscle of tibetan antelope significantly higher than that of tibetan sheep and low altitude sheep (P < 0.05). And the content of LA in skeletal muscle of tibetan antelope significantly lower than that of tibetan sheep and low altitude sheep (P < 0.05), activity of LDH and MDH in skeletal muscle was significantly lower and higher respectively than that of tibetan sheep and low altitude sheep (P < 0.05). There was no significant difference between tibetan sheep and low altitude sheep.

CONCLUSIONTibetan antelope may improve their ability to get oxygen under hypoxia by increasing the content of myoglobin in skeletal muscle, and the proportion of aerobic metabolism is high in skeletal muscle, it may be relate that with high myoglobin content in skeletal muscle, we suppose that high myoglobin content in skeletal muscle of tibetan antelope might be one of the molecular basis to adapt hypoxia.

Altitude ; Animals ; Antelopes ; metabolism ; physiology ; Hypoxia ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Malate Dehydrogenase ; metabolism ; Muscle, Skeletal ; metabolism ; Myoglobin ; metabolism

BACKGROUNDStreptococcus (S.) oligofermentans is a newly identified bacteria with a yet to be defined mechanism of sucrose metabolism that results in acid production. This study aimed to investigate the biochemical mechanisms of S. oligoferm-entans glucose metaolism.

METHODSThe S. oligofermentans LMG21532, Lactobacillus (L.) fermentum 38 and the S. mutans UA140 were used to characterize sucrose metabolism by measuring lactate dehydrogenase (LDH) activity and lactic acid production. Continuous dynamics and high performance capillary electrophoresis were used to determine LDH activity and lactic acid production, respectively, from bacteria collected at 0, 10 and 30 minutes after cultured in 10% sucrose.

RESULTSThese analyses demonstrated that LDH activity of the three bacterial strains examined remained stable but significantly different throughout the sucrose fermentation process. The S. oligofermentans LDH activity ((0.61 ± 0.05) U/mg) was significantly lower than that of L. fermentum ((52.91 ± 8.97) U/mg). In addition, the S. oligofermentans total lactate production ((0.048 ± 0.021) mmol/L) was also significantly lower than that of L. fermentum ((0.958 ± 0.201) mmol/L). Although the S. oligofermentans LDH production was almost double of that produced by S. mutans ((0.32 ± 0.07) U/mg), lactic acid production was approximately one sixth that of S. mutans ((0.296 ± 0.058) mmol/L). Additional tests examining pyruvic acid production (the LDH substrate) demonstrated that lactic acid concentrations correlated with pyruvic acid production. That is, pyruvic acid production by S. oligofermentans was undetectable following sucrose incubation, however, (0.074 ± 0.024) and (0.175 ± 0.098) mmol/L pyruvic acid were produced by S. mutans and L. fermentum, respectively.

CONCLUSIONS. oligofermentans is incapable of fermenting carbohydrates to produce enough pyruvic acid, which results in reduced lactic acid production.

L-Lactate Dehydrogenase ; metabolism ; Lactic Acid ; metabolism ; Pyruvic Acid ; metabolism ; Streptococcus ; metabolism ; Sucrose ; metabolism

AIMTo explore the adapting metabolic mechanisms of the plateau zokors to the hypoxic-hypercapnic environment.

METHODSThe activities of lactate dehydrogenase in serum and tissues, and the content of lactate in serum of plateau zokors in spring, summer and autumn were determined by using method of enzyme analysis. The spectrums of lactate dehydrogenase isoenzymes in serum and tissues of plateau zokors in spring, summer and autumn were analyzed by using method of the discontinuous systemic poly-acrylamide perpendicular plank gel electrophoresis.

RESULTSThe activities of lactate dehydrogenase in serum had obvious seasonally difference that were higher in spring and lower in autumn, and the content of lactate in serum showed same changing pattern. The spectrums of lactate dehydrogenase isoenzymes in serum showed five bands that were LDH1, LDH2, LDH3, LDH4 and LDH5 from positive pole to negative pole respectively, it showed clearly two bands in serum of summer that were LDH4 and LDH5 and one band in serum of autumn that was LDH5. The activities of LDH in tissues of skeleton muscle, cardiac muscle and brain were higher compared with the other tissues, it decreased markedly from spring to summer to autumn. In tissues of liver, kidney and lungs, activities of LDH were lower. Activities of LDH in livers, were significantly higher in spring compared that in summer and autumn, which had no obvious difference between summer and autumn. Activities of LDH in kidneys and lungs, showed no obviously difference between spring and summer, which decreased markedly in autumn. The spectrums of lactate dehydrogenase isoenzymes in tissues of cardiac muscle, liver, lungs, kidney, brain and skeleton muscle showed five bands, the spectrums were obvious different in different tissues, and the content of LDH isoenzymes showed seasonal changes in different tissues.

CONCLUSIONGlycolysis levels in plateau zokors had obvious seasonally change which increased in spring and decreased in autumn significantly. It related to the activity of plateau zokors in different seasons and seasonal fluctuation of oxygen and carbon dioxide in burrows of plateau zokors.

Animals ; Carbon Dioxide ; metabolism ; Isoenzymes ; analysis ; metabolism ; L-Lactate Dehydrogenase ; analysis ; metabolism ; Rodentia ; metabolism ; Seasons

The aim of the present study was to explore the changes and roles of dystrophin and membrane permeability in hypoxic training. Seventy-two 8-week-old Sprague Dawley (SD) rats were randomly divided into 4 groups, normoxic non-train (NC), normoxic train (NT), hypoxic non-train (HC), and hypoxic train (HT) groups. The rats of each group were randomly divided into three subgroups, non-exhaustive, low-speed exhaustive test and high-speed exhaustive test subgroups. Rats in hypoxia groups lived and were trained in a condition of 12.7% oxygen concentration (equal to the 4 300 m altitude). NT and HT groups received 4 weeks of training exercise. Then the rats in all non-exhaustive subgroups were sacrificed, and gastrocnemii were sampled for the measurements of lactate dehydrogenase (LDH), succinatedehydrogenase (SDH), malate dehydrogenase (MDH) activities. Moreover, serum LDH activity was analyzed. Low-speed exhaustive test and high-speed exhaustive test subgroups received exhaustive tests with 20 (71% VO2max) and 30 m/min speed (86% VO2max), respectively, and their exhaustive times were recorded. The results showed that, compared with normoxic groups, the weights in hypoxia groups exhibited slower increase. The level of dystrophin in HT group without exhaustion test didn't change significantly. The muscle MDH activities were markedly affected by the different oxygen concentration, training and their interaction (P < 0.05), whereas the muscle LDH activities were only affected by the different oxygen concentration (P < 0.05). Serum LDH activities were affected by the interaction of the different oxygen concentration and training (P < 0.05), showing decreased muscle LDH and increased blood LDH activities. The exhaustion time were markedly affected by the different test speed, training and their interaction (P < 0.05), and also affected by the interaction of the different oxygen concentration and training (P < 0.05), but didn't affected by oxygen concentration. The exhaustive time of HT high-speed exhaustive test subgroup was more than NT high-speed exhaustive test subgroup in 30 m/min exhaustion test. Compared with NT high-speed exhaustive test subgroup, HT high-speed exhaustive test subgroup had an earlier fatigue in the test, but had a rapid recovery. These results suggested that hypoxic training can effectively increase the rats' high-speed exhaustive time. The mechanism may be related to an increase in serum LDH caused by the increased membrane permeability after hypoxic training.
Altitude ; Animals ; Dystrophin ; metabolism ; Fatigue ; Hypoxia ; L-Lactate Dehydrogenase ; metabolism ; Malate Dehydrogenase ; metabolism ; Muscle, Skeletal ; enzymology ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Succinate Dehydrogenase ; metabolism

Lactic dehydrogenase (LDH) isozyme pattens were examined in rats after exposing the animal to 250 ppm of sulfur dioxide gas. The isozymes of the respective tissues were separated on cellulose-acetate strips from the brain, lung, heart, liver, kidneys, and muscle, and visualized as the isozyme bands by the formazan reaction and analyzed by densitometry. As well as the above experiment, room-air and room-air+SO2 were aerated through tissue homoenates in-vitro, accompanied by pure oxygen aeration in order to see the in vitro effect of the gases on the LDH activity in the tissues mentioned with the following conclusions. (1) The H-type of LDH activity dominated in the normal heart tissue of rats, M-type in the normal lung, liver, and muscle tissues of the animal. (2) The kidney tissue of normal rats exhibited preponderance of LDH-1 and-5 isozymes, brain tissue in LDH-1 and-4 isozymes. (3) When rats inhaled sulfur dioxide gas in the concentration of 250 ppm, it appeared that the M-type tended to predominate in the anaerobic tissues and the H-type in the aerobic tissue. (4) The degree of oxygen tension seemed to be correlated with the low level of LDH activity in the anaerobic tissues such as liver and muscle and with the increased activity in the aerobic tissues, such as heart and lung. (5) The low oxygen tension seems to favor syn-thesis of M-type LDH and high oxygen tesnion H-type LDH in the tissues of rats.
Air Pollution ; Animal ; Citric Acid Cycle ; Densitometry ; Electrophoresis ; Environmental Exposure ; Isoenzymes ; Lactate Dehydrogenase/metabolism* ; Oxygen/metabolism ; Rats ; Sulfur Dioxide/toxicity* ; Substances: ; Isoenzymes ; Sulfur Dioxide ; Oxygen ; Lactate Dehydrogenase

Plateau zokor (Myospalax baileyi) and plateau pika (Ochotona curzniae) are native to the Qinghai-Tibet plateau. To study their adaptive mechanisms, the ratios of heart weight to body weight (HW/BW) and right to left ventricular plus septum weights [RV/(LV+S)] were determined; the microvessel density (MVD) of cardiac muscle were measured by immunohistochemical staining; the numerical density on area (N(A)), volume density (V(V)), specific surface (δ), and surface density (S(V)) of mitochondria were obtained by microscopy and stereology; the contents of myoglobin (Mb) and lactic acid (LD), and the activity of lactate dehydrogenase (LDH) in cardiac muscle were analyzed by spectrophotometer. The results showed that the HW/BW of plateau zokor [(4.55±0.26)%] and plateau pika [(4.41±0.38)%] was significantly greater than that of Sprague-Dawley (SD) rat [(3.44±0.41)%] (P<0.05), but the RV/(LV+S) [(22.04±1.98)%, (25.53±3.41)%] was smaller than that of SD rats [(44.23±3.87)%] (P<0.05). The MVD and N(A) of cardiac muscle were 1688.631±250.253 and 0.768±0.123 in SD rat, 2002.888±367.466 and 0.868±0.159 in plateau pika and 2 990.643±389.888 and 1.012±0.133 in plateau zokor. The V(V) of mitochondria in plateau zokor (0.272±0.045) was significantly lower than that in plateau pika (0.343±0.039) and SD rat (0.321±0.048) (P<0.05), while the δ of mitochondria in plateau zokor (9.409±1.238) was higher than that in plateau pika (6.772±0.892) and SD rat (7.287±1.373) (P<0.05). The S(V) of mitochondria in plateau pika (2.322±0.347) was not obviously different from that in plateau zokor (2.468±0.380) and SD rat (2.227±0.377), but that in plateau zokor was significantly higher than that in SD rat (P<0.05). The contents of Mb in cardiac muscle of plateau zokor [(763.33±88.73) nmol/g] and plateau pika [(765.96±28.47) nmol/g] were significantly higher than that of SD rat [(492.38±72.14) nmol/g] (P<0.05), the content of LD in plateau zokor [(0.57±0.06) mmol/L] was obviously higher than that in plateau pika [(0.45±0.06) mmol/L] and SD rat [(0.48±0.02) mmol/L] (P<0.05), and the activity of LDH in plateau zokor [(16.90±2.00) U/mL] and plateau pika [(20.55±2.46) U/mL] were significantly lower than that in SD rat [(38.26±6.78) U/mL] (P<0.05). The percentage of LDH-H in cardiac muscle decreased in order in plateau zokor, plateau pika and SD rat. In conclusion, plateau zokor and plateau pika adapt better to hypoxia than SD rat by increasing the SV of mitochondria, MVD and content of Mb in the cardiac muscle. However, the parameters of mitochondria in the two high-altitude animals are different possibly because of the differences of habitats and habits.
Adaptation, Physiological ; Animals ; Heart ; physiology ; Hypoxia ; metabolism ; Isoenzymes ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Lagomorpha ; physiology ; Rats ; Rats, Sprague-Dawley ; Tibet

OBJECTIVE: To examine the effects of cocaine on the activities of ATPase, LDH and SDH in cultured mouse splenocytes in vitro. METHODS: The ATPase, LDH and SDH activities in mouse splenocytes were detected at day 7 after continuous culturing the mouse cells exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL in vitro. RESULTS: The activities of ATPase, LDH and SDH in mouse splenocytes exposed to cocaine hydrochloride in final concentration of 10, 20 and 100 microg/mL were significantly decreased after continuous culturing for 7 days. CONCLUSION The present study demonstrated that cocaine could inhibit the activities of ATPase, LDH and SDH in cultured splenocytes in vitro.
Adenosine Triphosphatases/metabolism* ; Animals ; Cells, Cultured ; Cocaine/pharmacology* ; Dose-Response Relationship, Drug ; L-Lactate Dehydrogenase/metabolism* ; Male ; Mice ; Mice, Inbred Strains ; Spleen/enzymology* ; Succinate Dehydrogenase/metabolism*

Albumin is the most abundant protein in human serum. A dye-binding method is commonly used in clinical laboratories for its estimation using different types of dyes. However, all these dye methods were interfered by a variety of compounds. Here we present a method for the detection of albumin in human serum and other biological fluids. The principle is based on the fact that lactate dehydrogenase isoenzyme-5 (LDH-5) binds specifically to Dextran-Blue (DB). Albumin inhibits the binding of LDH-5 with DB. Absence of LDH activity in DB fraction after gel filtration indicates the presence of albumin in sample and vice versa.
Chemistry, Clinical/*methods ; Chromatography, Gel ; Human ; Isoenzymes/metabolism ; Lactate Dehydrogenase/metabolism ; Protein Binding ; Sepharose/chemistry ; Serum Albumin/*analysis

OBJECTIVETo investigate the effect of lemon peel extracts (LPE) on the activity of lactate dehydrogenase and sucrase of Streptococcus mutans (Sm).

METHODSAfter serial dilution with trypticase soy broth (TSB) medium containing 2% glucose, LPE was used as the experimental group, and TSB without LPE as the control group. Sm was added to each group, which was then cultured for 6, 18, 24 and 48 hours in the anaerobic tank. The activity of lactate dehydrogenase(LDH) was measured with the method of oxidation of reduction coenzymeIand the pH value of the culture solution was also detected. The activity of the sucrose was determined with the method of coloration of 3,5-dinitrosalicylic acid.

RESULTSThe activity of LDH, sucrase and the changes of solution pH were decreased with the increase of the concentration of LPE (P < 0.01). The activity of LDH were declined from (0.8025 ± 0.0913) × 10(3) U/L to (0.2099 ± 0.0283) × 10(3) U/L; the activity of sucrase were declined from (-0.0107 ± 0.0003) × 10(3) U/L to (-0.0078 ± 0.0002) × 10(3) U/L; the ΔpH were declined from (2.8067 ± 0.0404) to (2.5033 ± 0.0416) (24 h results). The differences were significant between experimental groups and the control group (P < 0.01), and there were also significant differences among experimental groups with different LPE concentration (P < 0.01). The inhibitory effect of acid generation and lactate dehydrogenas' activity of Sm were positively correlated (P < 0.01).

CONCLUSIONSLPE can inhibit the activity of lactate dehydrogenase, sucrase and the acid production capacity of the Sm in a dose dependent manner. The inhibitory effects in logarithmic phase is stronger than that in other phases of growth cycle.

Citrus ; chemistry ; Glucose ; L-Lactate Dehydrogenase ; metabolism ; Lactic Acid ; Plant Extracts ; pharmacology ; Streptococcus mutans ; enzymology ; Sucrase ; metabolism

To improve the productivity of L-phenyllactic acid (L-PLA), L-LcLDH1(Q88A/I229A), a Lactobacillus casei L-lactate dehydrogenase mutant, was successfully expressed in Pichia pastoris GS115. An NADH regeneration system in vitro was then constructed by coupling the recombinant (re) LcLDH1(Q88A/I229A) with a glucose 1-dehydrogenase for the asymmetric reduction of phenylpyruvate (PPA) to L-PLA. SDS-PAGE analysis showed that the apparent molecular weight of reLcLDH1(Q88A/I229A) was 36.8 kDa. And its specific activity was 270.5 U/mg, 42.9-fold higher than that of LcLDH1 (6.3 U/mg). The asymmetric reduction of PPA (100 mmol/L) was performed at 40 °C and pH 5.0 in an optimal biocatalytic system, containing 10 U/mL reLcLDH1(Q88A/I229A), 1 U/mL SyGDH, 2 mmol/L NAD⁺ and 120 mmol/L D-glucose, producing L-PLA with 99.8% yield and over 99% enantiomeric excess (ee). In addition, the space-time yield (STY) and average turnover frequency (aTOF) were as high as 9.5 g/(L·h) and 257.0 g/(g·h), respectively. The high productivity of reLcLDH1(Q88A/I229A) in the asymmetric reduction of PPA makes it a promising biocatalyst in the preparation of L-PLA.
L-Lactate Dehydrogenase ; genetics ; Lactobacillus casei ; enzymology ; genetics ; Phenylpyruvic Acids ; metabolism ; Pichia ; genetics ; Recombinant Proteins ; genetics ; metabolism