• Title/Summary/Keyword: Lactate dehydrogenase (LDH)

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Purification and Characterization of Lactate Dehydrogenase Isozymes in Channa argus (가물치(Channa argus) 젖산탈수소효소 동위효소들의 정제 및 특성)

  • Park, Eun-Mi;Yum, Jung-Joo
    • Journal of Life Science
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    • v.20 no.2
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    • pp.260-268
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    • 2010
  • The lactate dehydrogenase (EC 1.1.1.27, LDH) isozymes in tissues from Channa argus were purified and characterized by biochemical, immunochemical and kinetic methods. The activity of LDH in skeletal muscle was the highest at 380.4 units and those in heart, eye and brain tissues were 13.4, 3,5 and 5.4 units, respectively. Citrate synthase (EC 4.1.3.7, CS) activity in heart tissue was the highest at 20.7 units. LDH/CS in skeletal muscle, heart, eye and brain tissues were 172.9, 0.6, 0.32 and 0.47. Protein concentration in skeletal muscle tissue was 14.7 mg/g and specific activities of LDH in skeletal muscle, heart, eye and brain tissues were 25.88, 0.79, 0.31 and 1.38 units/mg, respectively. Therefore, skeletal muscle tissue was anaerobic and heart tissue was aerobic. The LDH isozymes in tissues were identified by polyacrylamide gel electrophoresis, immunoprecipitation and Western blot with antiserum against $A_4$, $B_4$, and eye-specific $C_4$. LDH $A_4$, $A_3B$, $A_2B_2$. $AB_3$ and $B_4$ isozymes were detected in every tissue, $C_4$, $AC_3$, $A_2C_2$ and $A_3C$ were detected in eye tissue, and $A_3C$ was found in brain tissue. LDH $A_4$, $A_3B$, $A_2B_2$, $AB_3$, $B_4$, eye-specific $C_4$ isozymes were purified by affinity chromatography and Preparative PAGE Cells. The LDH $A_4$ isozyme was purified in the fraction from elution with $NAD^+$ containing buffer of affinity chromatography. Eye-specific $C_4$ isozyme was eluted right after $A_4$, after which $B_4$ isozyme was eluted with plain buffer. As a result, one part of molecular structures in $A_4$, $B_4$ and eye-specific $C_4$ were similar, but were different from each other in $B_4$ and $C_4$. Therefore the subunit A may be conservative in evolution, and the evolution of subunit B seems to be faster than that of subunit A. The activity of LDH $A_4$, $A_2B_2$, $B_4$, and eye-specific $C_4$ isozymes remained at 39.98, 21.28, 19.67 and 16.87% as a result of the inhibition by 10 mM of pyruvate, so the degree of inhibition was very high. The $Km^{PYR}$ values were 0.17, 0.27 and 0.133 mM in $A_4$, $B_4$ and eye-specific $C_4$ isozymes, respectively. The optimum pH of LDH $A_4$, $B_4$, eye-specific $C_4$, $A_2B_2$, $A_3B$, and $AB_3$ were pH 6.5, pH 8.5, pH 5.5, pH 6.0-6.5, pH 5.0 and pH 7.5. The $A_4$ and heterotetramer isozymes stabilized a broad range of pH. Especially, LDH activities in skeletal muscle tissue were high, resulting in a high degree of muscle activity.LDH metabolism in eye tissue seems to be converted faster from pyruvate to lactate by eye-specific $C_4$ isozyme as eye-specific $C_4$ have the highest affinity for pyruvate, and right after the conversion, oxidation of lactate was induced by $A_4$ isozyme. It was found that expression of Ldh-C, affinity to substrate and reaction time of $C_4$ isozyme were different according to the ecological environmental and feeding capturing patterns.

The effect of aronia, montmorency, and Prunus mume supplementation on blood fatigue factors and muscle damage enzymes after running 10 km

  • Shin, Pan-Jun;Kim, Yeon-Hui;Lim, In-Soo
    • Korean Journal of Exercise Nutrition
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    • v.16 no.1
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    • pp.19-26
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    • 2012
  • The purpose of this study was to analyze the effects of intake of aronia, montmorency, and Prunus mume on 10 km endurance exercise by investigating changes in blood lactate, ammonia, creatine kinase (CK), and lactate dehydrogenase (LDH), and eventually to recommend them as ergogenic aids, which are effective for recovering from exercise fatigue and muscle damage. The subjects were divided into a P. mume intake group, an aronia intake group, a montmorency intake group, and a control group. Blood samples were taken from the brachial vein at rest, and at 0, 30 min, and 24 hr after running to measure recovery status. All data were analyzed by a two-way repeated-measures analysis of variance using SPSS. The results are summarized as follows: The changes in blood ammonia and LDH concentration following 10 km of running were not significantly different among the groups. However, lactate and CK recovery ratio increased significantly in the aronia and montmorency groups compared to those in the P. mume and control groups. In particular, CK concentration was significantly different in the montmorency group compared to that in the other groups. These results suggest that the effects of aronia and montmorency are associated with improved recovery from muscle fatigue and damage induced by highly intensive exercise, endurance exercise, or overtraining. In particular, montmorency is recommended as an ergogenic aid to inhibit muscle pain resulting from high-intensity endurance exercise.

Study on Biochemical Pollutant Markers for Diagnosis of Marine Pollution III. Changes in Cholinesterase Activity of Flounder(Paralichthys olivaceus)in the Yellow Sea (해양오염의 진단을 위한 생화학적 오염지표에 관한 연구 III. 황해산 넙치(Paralichthys olivaceus)의 콜린에스테라아제 활성의 변화)

  • Choi, Jin-Ho;Kim, Dong-Woo;Moon, Young-Sil;Park, Chung-Kil;Yang, Dong-Beom
    • Journal of Life Science
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    • v.7 no.1
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    • pp.17-23
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    • 1997
  • This study was designed as a part of efforts to investigate the biochemical pollutant markers for diagnosis of maine pollutions by changes in cholinesterase activity of the flounder (Paralichthys olivaceus)in Yellow Sea of Korea. Acetylcholinesterase (AChE) activities in brain and muscle of cultured flounders in Yellow Sea were remarkably lower (40-50% and 40-55%, respectively)than those of wild flounder in Pohang (control) of East Sea, but AChE activities in brain and muscle of wild flounders in Yellow Sea were significantly lower(15-40% and 25-35%, respectively)than those of wild flounder in Pohang of East Sea. Butyrylcholinesterase(BChE) activities in barin and muscle of cultured flounders in Yellow Sea were remarkably lower(70-75% and 65-75%, respectively) than those of wild flounder in Pohang of East Sea, but BChE activities in barin and muscle of wild flounders in Yellow Sea were significantly lower (15-40%and 25-35%, respectively)than those of wild flounder in Pohang of East Sea. Lactate dehydrogenase (LDH) activities in serum of cultured flounders in Yellow Sea were significantly 10-50% higher than those of wild flounder in Pohang of East Sea, but LDH activities in serum of wild flounders in Yellow Sea were significantly 20-25% higher than those of wild flounder in Pohang of East Sea. It suggests that AChE and BChE activities in brain and muscle of cultured and wild flounders of Yellow Sea may be used as the most effective mean in a biochemical markers for diagnosis of pollutant effects by organophosphorus pesticides.

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Lactate Dehydrogenase and Monocarboxylate Transporters 1, 2, and 4 in Tissues of Micropterus salmoides (큰입우럭(Micropterus salmoides) 조직의 젖산탈수소효소 및 Monocarboxylate 수송체(MCT) 1, 2, 4)

  • Yum, Jung-Joo;Yeon, Jun-Hee
    • Journal of Life Science
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    • v.22 no.1
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    • pp.98-109
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    • 2012
  • The properties of lactate dehydrogenase (EC 1.1.1.27, LDH) and expression of monocarboxylate transporters (MCTs) 1, 2, and 4 were studied in tissues from Micropterus salmoides. Native-PAGE revealed that the LDH $A_4$ isozyme was predominantly located in skeletal muscle. The LDH $A_4$, $A_2B_2$, and $B_4$ isozymes were detected in heart, liver, eye, and brain tissues, while eye-specific $C_4$ isozyme was detected in eye tissue. In September, strong LDH $B_4$ isozyme activity was detected in heart tissue. High $A_4$ isozyme activity was noted in all other tissues except heart tissue. However, in November, strong $A_4$ isozyme activity was detected in heart tissue. The LDH/CS (Citrate synthase, EC 4.1.3.7) ratio in skeletal muscle and heart tissues indicated that anaerobic metabolism was high in those tissues. Native-PAGE after immunoprecipitation showed that eye-specific $C_4$ isozyme was more similar to the $A_4$ than the $B_4$ isozyme. The LDH $A_4$ isozyme was purified by affinity chromatography. The molecular weight of subunit A was 37,200. The LDH activity in tissues was consistently 11.05~28.32% due to inhibition by 10 mM pyruvate. The $K_m^{PYR}$ of LDH in eye tissue was very low. The optimum pH for LDH in tissues was pH 7.5~8.0. The LDH $A_4$ isozyme was detected in mitochondria of skeletal muscle, whereas the $B_4$ and $A_2B_2$ isozymes were detected in heart tissue mitochondria. Western blot analysis indicated that MCTs 1, 2, and 4 were located in the plasma membrane and mitochondria of skeletal muscle and heart tissues. The sizes of MCTs 1, 2, and 4 in skeletal muscle were 60, 54~38, and 63 kDa, while those in heart tissue were 57, 54~38, and 55.5 kDa, respectively. In conclusion, M. salmoides appears to use anaerobic metabolism predominantly when adapted to a hypoxic environment. In highly activated skeletal muscle and heart tissue, energy production is controlled by inward and outward flows of pyruvate and lactate through MCTs 1, 2, and 4 in the plasma membrane and mitochondria, with effective adjustment by LDH isozymes.

Comparative Analysis of the Pre-Post a Treadmill Exercise on the Blood Lactate, LDH and Glucose in the Runners (육상선수들의 트래드밀운동 전 ${\cdot}$ 후 Blood Lactate, LDH, Glucose의 비교 분석)

  • Park, Han-Su;Shin, Byung-Cheul;Chae, Jeong-Ryong;Jo, Sung-Cho;Jeon, Hee-Young;Kim, Hyunng-Jun;Kim, Mo-Kyung
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.5
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    • pp.1254-1260
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    • 2006
  • The purpose of this study was to the Pre-Post submaximal exercise for analyze the Blood Lactate, LDH(Lactate Dehydrogenase) and Glucose in the runners. The subjects were 12 volunteers 6 sprinters and 6 distance runners who agreed to intention of this study. Subjects peformed until possible all-out trials. Blood samples were taken from a Ante cubital vein Pre-Post exercise and every rest 5min during the all-out trial. The results obtained were summarized as follow; 1) Blood Lactate of Pre-Post exercise in treadmill test showed no significant difference between sprinters and distance runners, but showed significant in sprinters and distance respectively. 2) Serum LDH of Pre-Post exercise in treadmill test showed no significant difference between sprinters and distance runners, and that showed no significant in sprinters and distance respectively. 3) Blood Glucose of Pre-Post exercise in treadmill test showed no significant difference between sprinters and distance runners, and showed no significant in distance, but showed significant in sprinters. These data indicate first that the value of 4mmol/${\ell}$, commonly referred to as OBLA(Onset of Blood Lactate),may often underestimate the upper limit of tolerance to lactate during a maximal endurance peformance test until all-out. second, our date suggested that the sprinters exercise decreases serum LDH activities and the distance increases serum LDH activities. therefore serum LDH concentration changed following a different exercise. Third. for the concentration of the glucose in blood the sprinters was show to be higher. However it needs more studies to find out the relationship between this result and the fitness factor. therefor, since the runners has an intermittent characteristic which includes a frequent momentary change, it needs the fitness training that being consisted of various training. in the training method, it needs the training process which is from the whole fitness to specific fitness to improve general physical ability.

Effects of Hypoxia on Root Growth and Anaerobic Fermentative Enzymes in Winter Cereal Seedlings (저산소 조건하에서 맥류 유묘의 근생장 및 혐기발효 효소의 반응)

  • Park Myoung Ryoul;Lim Jeong Hyun;Yoo Nam Hee;Kwon In Sook;Kim Jung Gon;Choi Kyung Gu;Yun Song Joong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.6
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    • pp.400-405
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    • 2005
  • Wet-injury often occurs in upland cereals growing in the paddy field due to oxygen deficiency in the rhizosphere caused by excessive water in the soil. Under hypoxia, energy metabolism is diminished causing non­reversible damage to root cells. This study was conducted to investigate effects of hypoxia on root growth and enzymes involved in the fermentative energy metabolism in upland cereals including barley, wheat, rye and triticale. Young seedlings were subject to hypoxia for up to 7 days. Root fresh weight and dry weight were decreased significantly by hypoxia for 5 to 7 days in all cereal seedlings. Root growth retardation under hypoxia was lowest in barley. Hypoxia-induced increases in activity and isozyme expression of alcohol dehydrogenase (ADH) and lactate dehydrogenase (LDH) were commonly observed in roots of all cereal seedlings. The inherent ADH activity levels were higher in barley but the hypoxia-induced increases in ADH activities were lowest in barley than other cereals. The inherent LDH activity levels were lower in barley and the hypoxia-induced increases in LDH activities were lower in barley than other cereals. The results suggest the importance of the rapid enhancement of fermentative enzyme systems for increased tolerance to hypoxia.

Effects of Gamibojungikki-tang on LDH activity of Cultured Spinal Sensory Neurons (가미보중익기탕이 배양 척수감각신경세포의 LDH 활성도에 미치는 영향)

  • Lee Chang Ha;Kwan Kang Beam;Park Jun Su;Song Yang Sun;Ryu Do Gen
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.2
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    • pp.343-347
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    • 2002
  • In order to darify the neuroprotective effect of Gamibojungikki-tang(GBJIKT) water extract on cultured mouse spinal sensory neuron damaged by glucose Oxidase (GO), NR (Neutral Red) assay and LDH (Lactate Dehydrogenase) activity assay were carried out after the cultured mouse spinal sensory neuron were preincubated with various concentrations of GBJIKT water extract for 3 hours prior to exposure of GO. Cell viability of cultured mouse spinal sensory neurons exposed to various concentrations of GO for 8 hours was decreased in a dose-dependent manner. NR/sub 50/ values were 50 mU/ml GO. Cultured mouse spinal sensory neurons in the medium containing various concentration of GO for 8 hours showed increasing of LDH activity. We knew that GO was toxic on cultured spinal sensory neurons. Pretreatment of GBJIKT water extract for 3 hours following GO prevented the GO-induced neurotoxicity such as increasing of LDH activity. These results suggest that GO shows toxic effect on cultured spinal sensory neurons and GBJIKT water extract is highly effective in proecting the neurotoxicity induced by GO.

웅성 랫트에서 이염화메탄의 사염화 탄소 독성 증폭효과

  • 김대병;김영철
    • Toxicological Research
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    • v.9 no.2
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    • pp.253-262
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    • 1993
  • The effects of dichloromethane (DCM) on carbon tetrachloride (CT) toxicity were examined in adult male rats. A concomitant treatment of rats with DCM (0.3, 0.6, 1.2 g/kg, po) significantly potentiated the hepatotoxicity of CT (1.0 g/kg, po) as determined by increase in serum GPT (glutamic pyruvic transaminase), GOT (glutamic oxaloacetic transaminase), and SDH (sorbitol dehydrogenase) activity 24 hr following the treatments. Serum LDH (lactate dehydrogenase) activity was increased by either DCM or CT treatment.

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Changes in Mitogen-activated Protein Kinase Activities During Acidification-induced Apoptosis in CHO Cells

  • Kim, Jin-Young;Jeong, Dae-Won;Roh, Sang-Ho;Min, Byung-Moo
    • International Journal of Oral Biology
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    • v.30 no.3
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    • pp.85-90
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    • 2005
  • Homeostatic pH is very important for various cellular processes, including metabolism, survival, and death. An imbalanced-pH might induce cellular acidosis, which is involved in many abnormal events such as apoptosis and malignancy. One of several factors contributing to the onset of metabolic acidosis is the production of lactate and protons by lactate dehydrogenase (LDH) in anaerobic glycolysis. LDH is an important enzyme that catalyzes the reversible conversion of pyruvate to lactate. This study sought to examine whether decreases in extracellular pH induce apoptosis of CHO cells, and to elucidate the role of mitogen-activated protein kinases (MAPKs) in acidification-induced apoptosis. To test apoptotic signaling by acidification we used CHO dhfr cells that were sensitive to acidification, and CHO/anti-LDH cells that are resistant to acidification-induced apoptosis and have reduced LDH activity by stable LDH antisense mRNA expression. In the present study, cellular lactic acid-induced acidification and the role of MAPKs signaling in acidification-induced apoptosis were investigated. Acidification, which is caused by $HCO{_3}^-$-free conditions, induced apoptosis and MAPKs (ERK, JNK, and p38) activation. However, MAPKs were slightly activated in acidic conditions in the CHO/anti-LDH cells, indicating that lactic acid-induced acidification induces activation of MAPKs. Treatment with a p38 inhibitor, PD169316, increased acidification-induced apoptosis but apoptosis was not affected by inhibitors for ERK (U0126) or JNK (SP600125). Thus, these data support the hypothesis that activation of the p38 MAPK during acidification-induced apoptosis contributes to cell survival.