• Title/Summary/Keyword: Isoenzyme

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Studies on the Serum Total Activities and Isoenzyme PAtterns of LDH in Non-Human Primates Reared in Korea (國內詞育 원숭이의 血淸 LDH의 總活性値와 isoenzyme에 관한 硏究)

  • 윤상보;김덕환;서지민;신남식;현병화;김명철;윤효인;박배근;송희종
    • Journal of Veterinary Clinics
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    • v.18 no.4
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    • pp.380-389
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    • 2001
  • Non-human primates have been increasing in demand as important experimental animals and companion animals, domestically and internationally. The number of non-human primates for these purposes will be much enhanced in the near future. Despite this trend, basic physiological data are scarcely available in these animal species, leading to the difficulty to diagnose diseases when necessary, due to the absence of reference values. Particularly, there is not any report on the total activity of LDH of non-human primates, let alone LDH isoenzyme patterns, in Korea. LDH isoenzymes have a high level of efficaciousness as diagnostic and prognostic aids in various diseases. In this study, total activities and isoenzyme patterns of LDH were measured to obtain their reference values in domestically reared common marmosets, crab-eating macaques and Japanese macaques. There were widespread different values of serum total LDH among the non-human primate species experimented in this study. Serum LDH values of common marmosets and crab-eating macaques were 597.5$\pm$243.1 IU/l and 605.3$\pm$312.6 IU/l, respectively, whereas those of Japanese macaque showed 1,209$\pm$473.8 IU/l. Five isoenzyme fractions of LDH were observed in all experimented non-human primates but their ranks and proportions represented different patterns one another. In common marmosets, the percent of fraction for serum LDH1, LDH$_2$, LDH$_3$, LDH$_4$, and LDH$_{5}$ was 13.7$\pm$6.4%, 23.3$\pm$3.6%, 29.2$\pm$5.0%, 9.4$\pm$1.4% and 24.4$\pm$7.5%, respectively. The rank of LDH isoenzymes was LDH$_3$>LDH$_{5}$>LDH$_2$>LDH$_1$>LDH$_4$, in the descending order. For crab-eating macaques, the fraction of serum LDH$_1$, LDH$_2$, LDH$_3$, LDH$_4$, and LDH$_{5}$ occupied 19.5$\pm$12.7%, 25.3$\pm$9.3%, 23.8$\pm$8.1%, 10.2$\pm$2.8% and 21.3$\pm$14.2%, respectively. The order of LDH isoenzymes was LDH$_2$>LDH$_3$>LDH$_{5}$>LDH$_1$>LDH$_4$, from top to down. On the while, in Japanese macaques, the fraction of serum LDH$_1$ to LDH$_{5}$ showed 23.4$\pm$11.8%, 30.5$\pm$4.1%, 17.4$\pm$3.9%, 11.3$\pm$3.7% and 13.8$\pm$5.6%, respectively. The decreasing order indicated LDH$_2$>LDH$_1$>LDH$_3$>LDH$_{5}$>LDH$_4$. In conclusion, values such as LDH and LDH isoenzyme patterns of investigated for the first time from non-human primates reaared in Korea, could be reference values for the optimal diagnosis and therapy of diseases of the corresponding animal species. Other parameters of hematology and blood biochemistry are urgently needed to study for the benefit of our intimate non-human primates.an primates.

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Activity Determination, Kinetic Analyses and Isoenzyme Identification of Gamma Glutamyltransferase in Human Neutrophils

  • Sener, Azize;Yardimci, Turay
    • BMB Reports
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    • v.38 no.3
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    • pp.343-349
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    • 2005
  • Gamma-glutamyltransferase (GGT, EC 2.3.2.2) which hydrolyzes glutathione (GSH), is required for the maintenance of normal intracellular GSH concentration. GGT is a membrane enzyme present in leukocytes and platelets. Its activity has also been observed in human neutrophils. In this study, GGT was purified from Triton X-100 solubilized neutrophils and its kinetic parameters were determined. For kinetic analyses of transpeptidation reaction, $\gamma$-glutamyl p-nitroanilide was used as the substrate and glycylglycine as the acceptor. Apparent $K_m$ values were determined as 1.8 mM for $\gamma$-glutamyl p-nitroanilide and 16.9 mM for glycylglycine. The optimum pH of GGT activity was 8.2 and the optimum temperature was $37^{\circ}C$. It had thermal stability with 58% relative activity at $56^{\circ}C$ for 30 min incubation. L-serine, in the presence of borate, was detected as the competetive inhibitor. Bromcresol green inhibited neutrophil GGT activity as a noncompetetive inhibitor. The neutrophils seem to contain only the isoenzyme that is present in platelets. We characterized the kinetic properties and compared the type of the isoenzyme of neutrophil GGT with platelet GGT via polyacrylamide gel electrophoresis (PAGE) under a standart set of conditions.

Effects of Temperature on the Isoenzymes of Peroxidase in Escherichia coli (Escherichia coli의 Peroxidase Isoenzyme에 미치는 온도의 영향)

  • 강사욱;강현삼;홍순우
    • Korean Journal of Microbiology
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    • v.14 no.1
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    • pp.39-47
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    • 1976
  • This experiment was designed to study the effects of temperature on the peroxidase isoenzymes of a mesophilic microorganism, Escherichia coli (grown within biokinetic zone). Optimum temperature for the growth of E. coli was $37{\circ}C.$ Three different temperatures, 20, 30 and $40{\circ}C,$ were selected. And the isoenzyme patterns of peroxidase of E. coli, growth respectively at each temperature, were analysed by disc electrophoresis. The sample of 20.deg.C showed 4 bands, that of 30.deg.C, 5 bands and that of 40.deg.C, 6 bands. Two dark bands (higher molecular weight, 56,000 and 54,000) and two light bands (lower molecular weight, 11,500 and 10,000) were constant at all samples. But two intermediate bands (M.W.44,000 and 34,000) were variable ; at $20{\circ}C,$ no banding pattern, one band 9M.W. 34,000) only at $30{\circ}C,$ and at $40{\circ}C$ two bands were appeared. And the shifts of growth temperatures between 30.deg.C and 40.deg.C showed the alteration of the isoenzyme patterns ; the isoenzyme patterns of the smaple of tmeperature shift from $30{\circ}C$ to $40{\circ}C$ were same as that of 40.deg.C and vice versa.

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Interstrain polymorphisms of isoenzyme profiles and mitochondrial DNA fingerprints among seven strains assigned to Acanthamoeba polyphaga (대식가시아메바(Acmthamoebapokphaga) 일곱 분리주간의 동위효소 profile과 Mitochondria DNA fingerprint의 다양성)

  • Gong, Hyeon-Hui;Park, Jun-Hyeong;Jeong, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.33 no.4
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    • pp.331-340
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    • 1995
  • Interstrain polymorphisms of isoenzyme profiles and mitochondrial (Mt) DNA fingerprints were observed among seven strains of Acnnthnmoeba isolated from different sources and morphologically assigned to A. polvphngn. Mt DNA ringerprints by eight restriction endonucleases (Bgl II, Sca I, Cla I, EcoR I, Xbo I, Kpn I, Sal I, and Sst I) revealed considerable interstrain polymorphisms . Isoenzyme profiles revealed considerable interstrain polymorphisms for acid phosphatase, lactate dehydrogenase, and glucose-6- phosphate dehydrogenase while those for glucose phosphate isomerase , leucine aminopeptidase , and malate dehydrogenase showed similarity Despite of the interstrain polymorphisms, the isoengyme profiles and Mt DNA fingerprints of the strain Ap were found to be identical with those of the strain .tones . Mt DNA fingerprinting was found to be highly applicable for the strain identification, characterization, and differentiation. Key words: Acanthnmoebn polyphcga, interstrain polymorphism, isoenzyme profiles , Mt DNA fingerprints, strain differentiation, strain identification.

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Lactate dehydrogenase activity and isoenzyme distribution in plasma and tissue of Korean native cattle (한우의 혈장 및 조직중의 lactate dehydrogenase의 활성치와 isoenzyme의 분포)

  • Kim, Ki-seog;Cho, Jong-hoo
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.461-467
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    • 1989
  • The activity of lactate dehydrogenase in plasma and various tissues(skeletal muscle, cardiac muscle, liver, lung, kidney and spleen) of Korean native cattle in a Chonju abattoir, the Breeding Stock Farm and Animal Farm of Chonbuk University was determined by using ultra violet method. Using polyacrylamide gel electrophoresis, the lactate dehydrogenase isoenzyme distrimution of plasma and various tissues in Korean native cattle was studied. The plasma lactate dehydrogenase activity of Korean native cattle was $554.80{\pm}92.70IU/l$ and the lactate dehydrogenase activity of male plasma was $543.96{\pm}97.89IU/l$, which was lower than that of female plasma, $579.19{\pm}78.09IU/l$. The plasma lactate dehydrogenase activity of calf was $557.31{\pm}110.27IU/l$ and was no significantly different from that of adult Korean native cattle. But the range of calf lactate dehydrogenase activity was larger than that of adult Korean native cattle. In tissues, the lactate dehydrogenase activity was decreased in order of lung, kidney, spleen, liver, heart and skeletal muscle. The lung had the greatest activity and the skeletal muscle had the least. Lactate dehydrogenase isoenzymes in plasma and tissues were found to have a characteristic distribution and quantitative isoenzyme patterns. In plasma, the LDH1 usually had the greatest activity and other isoenzymes showed a decreasing tendency in order of LDH2, LDH3, LDH4 and LDH5. The distribution of lactate dehydrogenase isoenzymes had a wide variation in tissues. But the distribution of LDH isoenzymes in plasma was similar to that in kidney, and also cardiac muscle and spleen had similar pattern in LDH isoenzymes distribution.

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1269S mutation in horse liver alcohol dehydrogenase S isoenzyme and its reactivity for steroids and retinoids

  • Ryu, Ji-Won;Lee, Kang-Man
    • Archives of Pharmacal Research
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    • v.20 no.2
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    • pp.115-121
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    • 1997
  • Ile-269 in horse liver alcohol dehydrogenase isoenzyme S(HLADH-S) was mutated to serine by phosphorothioate-based site-directed mutagenesis in order to study the role of the residue in coenzyme binding. The specific activity of the mutant(1269S) enzyme to ethanol was increased 49-fold. All turnover numbers of 1269S enzyme toward 9 primary alcohols were increased. The mutant enzyme showed 3.6, 4.6, 11.6-fold higher catalytic efficiency for $5{\beta}$-androstane-3, 17-dione, $5{\beta}$-cholanic acid-3-one and retinal than wild-type, respectively. The reaction mechanism of 1269S enzyme was ordered bi bi as wild-type's. These results indicate that the hydrophobic interaction of Ile-269 residue with coenzyme plays an important role in dissociation of coenzyme from enzyme-coenzyme complex, which has been known as the rate limiting step of ADH reaction.

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Creatine Kinase and its Isoenzymes in Jindo Dogs (진도견의 creatine kinase 활성치와 isoenzyme)

  • 최석화
    • Journal of Veterinary Clinics
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    • v.15 no.1
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    • pp.41-45
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    • 1998
  • This study was performed to investigate the serum creative kinase(CK) activity and CK isoenzyme in Jindo dog. Serum CK activity and CK-isoenzyme were analyzed in 53 Jindo dogs of both sexes. The mean value and normal range of serum CK activity were 24.1 lu/$\ell$ and 7-91 lu/$\ell$, respectively, in 29 female dogs, 24.8 lu/$\ell$ and 8-89 lu/$\ell$ in 24 male dogs. The CK activity of the Puppy showed a tendency to be higher than that of the adult. There was no significance between Puppy and adult. Three isoenzymes (CK-MM, CK- BB, and CK-MB) were recognized in serum. The mean percentages of female and male were as follows: 48.31fp and 48.1% for CK-MM, 35.49) and 33.61fp for CK-BB, and 8.2% and 10.1% for CK-MB in the puppy and 46.21% and 46.1 % for CK-MM, 36.3% and 37.6% for CK-BB and 10.5% and 9.5% for CK-MB in the adult.

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Diagnostic Significance of Total Lactate Dehydrogenase(LD) and LD Isoenzyme Measurement in the Body Fluids (체강액에서 총 Lactate dehydrogenase 및 Lactate dehydrogenase 동위효소 측정의 진단학적 의의)

  • Jeon, Chang-Ho;Bae, Eun-Kyung;Hong, Seok-Il;Kim, Chung-Sook;Lee, Young-Hyun
    • Journal of Yeungnam Medical Science
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    • v.3 no.1
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    • pp.193-199
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    • 1986
  • Body fluid Lactate dehydrogenase and its isoenzyme measurement was performed in 132 patients: 8 cases with peritonitis, 21 cases with malignant ascites, 43 cases with liver cirrhosis, 48 cases with tuberculous pleuritis, 12 cases with malignant pleural effusion respectively. Body fluid protein and glucose contents, red blood cell counts, white blood cell counts, cytologic examination were also performed as a comparative study. The results were as follows: 1. Measurement of total LD and protein amount could differentiate between transudate and exudate in the ascitic fluids. 2. In the malignant exudate of ascites and pleural fluid, the activity of LD2 isoenzyme was statistically increased compared with that of inflammatory exudate and the activity of LD4 isoenzyme was also increased compared with that of serum(P<0.05). 3. The inflammatory exudate of pleural fluid and ascites demonstrated the increase of LD5 isoenzyme activity stastistically compared with that of serum and malignant exudate(P<0.05). 4. A difference of total LD activity between malignant ascites and inflammatory ascites was significant statistically, while this was not observed in the pleural exudate. 5. Total LD and LD5 isoenzyme activity didn't correlated with the number of white blood cells in the exudate.

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Radioprotective Effect of Methylene Blue 1. Effect of Methylene Blue on Lactic Dehydrogenase Level and Lactic Dehydrogenase Isoenzymes of Rats Exposed to Gamma-Irradiation (Methylene Blue의 방사선 방어결과 1. Methylene Bluerk가 $\gamma$선에 조사된 흰쥐의 유산탈수소효소의 함량과유산탈수소효소의 동위효소에 미치는 영향)

  • Sang Yul Nam;Seung Han Chang
    • The Korean Journal of Zoology
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    • v.12 no.3
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    • pp.94-102
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    • 1969
  • The protective action of methylene blue against gamma-irradiation was studied with rats. Albino rats were given 360 rads of whole-body gamma-irradiation following an intraperitoneal injection of physiological saline or methylene blue. Male rats given methylene blue (38mg/kg) and the control rats given saline were alive following gamma-irradiation. Serum lactic dehydrogenase (LDH) activity, and LDH isoenzyme patterns in serum and various organs were determined at various time intervals after the exposure. 1) The serum LDH level in both the control and methylene blue-treated rats was increased during the initial phase, but returned to the initial level thereafter. 2) Methylene blue showed a marked delay in the rise of serum LDH at 15 and 64 hours after exposure. 3) The exposure in the control and methylene blue-treated rats resulted in an increase in the relative amount of the more electrophoretically mobile-anodal isoenzyme (band 1) and a decrease in the least mobile-cathodal isoenzyme (band 5) in serum, liver, heart and testis nearly at 40 and 116 hours, respectively. 4) Isoenzyme patterns in serum, liver and testis after exposure were not significantly different between the control and the methylene blue-treated rats. 5) Methylene blue showed a slight delay in alteration of heart tissue LDH isoenzyme patterns after exposure. 6) The increase of serum LDH level after exposure is a reflection of an immediate increase in the H type, band 1 of LDH isoenzymes. 7) It is concluded from this study that methylene blue has a remarkable radioprotective action in the serum LDH activity and in the heart tissue LDH isoenzyme patterns.

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Isoenzyme patterns and phylogenetic relationships in Acanthamoeba spp. isolated from contact lens containers in Korea

  • Shin, Ho-Joon;Cho, Myung-Soo;Kim, Han-jip;IM, Kyung-Il
    • Parasites, Hosts and Diseases
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    • v.37 no.4
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    • pp.229-236
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    • 1999
  • In order to refer to the basic information regarding the identification of isolates obtained from a contact lens container in Korea, the isoelectric focusing gel electrophoresis was employed to compare the isoenzyme band patterns among Acanthamoeba spp. including eight isolates and the simple pairwise dissimilarity analysis was carried out. For an alkaline phosphate development, isolate 7 and Acanthamoeba polyphaga showed homologous band patterns, and isolates 1, 2, and 3 showed the same patterns. For lactate dehydrogenase, similar patterns were observed in isolates 2 and 3. Isolates 3 and 5 showed homologous band patterns for malate dehydrogenase and glucose phosphate isomerase. For hexokinase, isolates 4, 7, and A. hatchetti showed the same band patterns. In others, a considerable number of interstrain polymorphisms was observed in nine isoenzyme band patterns. In Acanthamoeba group II, genetic distances among isolates 1, 2, 3, 4, and 5 ranged from 0.104 to 0.200. In comparison to A. castellanii, A. hatchetti, and A. poIyphaga, genetic distances of isolates 7 and 8 were 0.254 and 0.219, respectively. In Acanthamoeba group III, including A. culbertsoni, A. healyi, and A. royreba, isolate 6 had genetic distances which ranged from 0.314 to 0.336. Finally, when comparing to the six reference Acanthamoeba, it was possible to classify isolates 1, 2, 3, 4, and 5, as genetically close-related species and as independent species group. Furthermore, isolates 6, 7 and 8 were identified as independent species as well.

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