• Title/Summary/Keyword: isoenzymes

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Peroxidase Isozyme in Root Differentiation from Cultured Ginseng Root Explants (인삼 근절편 배양시 Peroxidase Isozyme에 관한 연구)

  • 김명원
    • Journal of Plant Biology
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    • v.29 no.4
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    • pp.233-242
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    • 1986
  • In order to pursue some physiological studies on organogenesis in ginseng tissue culture, ginseng root explants were cultured on a modified MS medium containing NAA and kinetin. The activities of peroxidase and some enzymes were investigated and their isoenzyme patterns were also observed. The activity of peroxidase decreased by 20% in one week's culture and increased thereafter by 80% in culturing for 7 weeks compared with the control group. Glucose-6-phosphate dehydrogenase activity increased by 400% after culturing for 5 weeks and increased during the days preceeding root formation. The activities of glutamate dehydrogenase and acid phosphatase also increased during the culture. After 3 weeks' culture, new peroxidase isozyme (pH 7.6) appeared and 7 weeks' culture, another new peroxidase isozyme (pH unidentified) appeared. These patterns were also identified by using FPLC. After 7 weeks' culture, a new esterase isozyme of pH 8.5 appeared and isozyme patterns of acid phosphatase were quite changed compared with the isozyme patterns of tissue cultured for 5 weeks. In so far as these new isoenzymes appear distinctively after 7 weeks' culture, root differentiation is supposed to be induced after 7 weeks' culture.

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Effect of 2,4-Dichlorophenoxyacetic Acid and Kinetin on Peroxidase Isoenzymes in Ginseng(Panax ginseng C. A. Meyer) Callus Cultures (고려인삼(Panax ginseng C.A.Meyer) 조직배양에서 2.4-dichloro phenoxyacetic acid와 kinetin첨가가 Isoperoxidase 변이에 미치는 영향)

  • 김명원;강영희
    • Journal of Ginseng Research
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    • v.7 no.1
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    • pp.52-62
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    • 1983
  • This study was undertaken to investigate the influence of kinetin and 2, 4-dichlorophenoxyacetic acid on the rate of growth, the contents of RNA, DNA, and protein. And also the effect of plant growth regulator on isoperoxidases in callus derived from root (root-callus) and petiole (petiolecallus) was investigated. The rate of growth in petiole-callus was higher than the rootcallus at 0.1 mg/l kinetin and 1mgfl 2,4-D. At 1mgll kinetic, the rate of growth increased, but at high concentration the rate of growth decreased fast. The contents of RNA, DNA and protein also increased, but it did not coincide with the increase of the growth rate of callus. The isoperoxidases of callus grown at various amounts of 2,4-D and kinetic occurred in an almost fashion, but those of root-callus appeared different from those of petiole-callus.

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The Variation of Isoenzymes and Morphological Characteristics of Needle, Cone and Seed According to Resin Duct Index in Pinus densiflora and Pinus thunbergii (소나무 및 곰솔의 수지구지수(樹脂溝脂數)에 따른 침엽(針葉), 구과(毬果) 및 종자(種子)의 형태적(形態的) 특성(特性)과 동위효소(同位酵素)의 변이(變異))

  • Son, Doo Sik;Park, Sang Jun;Hwang, Jae Woo
    • Journal of Korean Society of Forest Science
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    • v.79 no.4
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    • pp.424-430
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    • 1990
  • The variation of needle, cone and seed characteristics and of allelic frequency in isoenzyme, ADH, ME and PGI, according to different resin duct index in Pinus densiflora, Pinus thunbergii and their hybrids was analyzed. The results obtained were as follows : 1. With increase of number of resin ducts, morphological characteristics such as needle length, needle sheath length, cone size, seed size, seed wing size, 1000 seeds weight, etc. tended to he increased, while number of stomata row in needle to be decreased. 2. As the results of discriminant analysis for the morphological characteristics of needle, cone and seed, most individuals are generally coincided with number of resin duct mostly in Pinus densiflora were not. 3. According to the canonical discriminant function obtained from the morphological characteristics in Pinus densiflora, Pinus thunbergii and their hybrids including introgressive hybrid, the resin duct index, 1000 seeds weight, cone size and neelde sheath length characterized fairly their species. 4. With increase of resin duct index, hybrid index tended to be higher. The results obtained from the discriminant analysis and the hybrid index were nearly same each other. 5. With increase of number of resin duct, the allelic frequencies for isoenzyme, ADH-$B_2$, ME-$A_2$ and PGI-$B_1$. $B_2$ tended to increase but those of ADH-$B_3$, ME-$A_4$, and PGI-$B_3$, to decrease. It is thought that this increase of frequency for the former four isoenzymes was resulted in introgressive gene flow from Pinus thunbergii to Pines densiflora and accordingly the frequency of latter three isoenzymes tended to decrease.

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Study on the Pattern of Isoenzymes in Pancreatic Juice, Serum and Saliva of Rabbit (정상 및 병적체액중 동종효소분획에 관한 연구)

  • Kim, Won-Joon;Kim, Hea-Young;Lee, Hyang-Woo;Hong, Sa-Suk
    • The Korean Journal of Pharmacology
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    • v.16 no.2 s.27
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    • pp.15-24
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    • 1980
  • [${\alpha}$]-Amylase catalyses the hydrolysis of starch, glycogen, and related poly- and oligosac-charide by random cleavage of ${\alpha}$-D-(l-4) glucan linkage. In man large amounts of amylase are secreted into the digestive tract by the salivary and exocrine pancreatic gland, minimal amount being produced also in other tissues. It has been known that ${\alpha}$-amylase exists in multiple molecular forms, isoenzyme which can be separated from each other because of difference in their physicochemical properties. By using various methods, several groups of investigator have separated the many isoenzyme in serum, saliva and pancreatic juice. Furthermore, changes of the normal serum isoenzyme pattern is diagnostically useful even when the total serum enzyme activity is noninformative, such as the clinical use of isoenzyme of serum lactate dehydrogenase. Procarboxypeptidase-A which is one of the pancreatic enzymes is also present as isoenzymes. Four forms of procarboxypeptidase-A haye been found in the bovine enzyme and three forms of the porcine enzyme. In human pancreatic juice four forms of procarboxypeptidase-A isoenzyme were found by isoelectric focusing method. Recently, the so-called isoamylase analysis was developed for the diagnostic use of amylase in pancreatic diseases. In alcohotic patients, the serum concentration of pancreatic isoamylase is subnormal and this lowered activity provides strong evidence for pancreatic exocrine insufficiency. The purpose of this study was to elucidate the variations of the isoenzyme of amylase and procarboxypeptidase-A in serum, saliva and pancreatic juice of the experimental animals. The results are as follow. 1) Three main forms of isoenzyme of amylase by isoelectric focusing were found in pancreatic juice of normal rabbit. However, many new bands were appeared in the pancreatic juice of cholic acid administered animal intravenously while the infusion of cholic acid or elastase into pancreatic duct produced the decrease of number of the fractions on the isoelectric focusing. In the case of serum isoenzyme from normal animal, two major and a few minor isoamylases were observed. By injecting alcohol intravenousely the fractions of serum isoamylase were significantly decreased and in contrary to the pattern in the pancreatic juice the infusion of cholic acid or elastase into pancreatic duct exhitited a significant decrease of the isoenzyme of amylase fractions. In saliva from normal animal three main isoamylase were produced of the administration of alcohol. 2) In the case of procarboxypeptidase-A isoenzyme, two major fractions which have isoelectric point at 6.2 and 6.4 and other two minor bands were observed in the pancreatic juice of normal rabbit. By the treatment of the juice with trypsin, only one band was produced on the isoelectric focusing. No procarboxypeptidase was appeared on the electrofocusing by the infusion of cholic acid or phospholipase A into the pancreatic duct of rabbit. However, a single major fraction of procarboxypeptidase-A was appeared at 3 hr after simple ligation of the pancreatic duct. No significant changes were observed in the juice of the alcohol or cholic acid administered group.

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Characterization of the Expression of PKCα(Isoform) in DMH-induced Vascular Endothelial Proliferation (DMH에 의한 비정상적인 혈관 내피세포의 증식에서 Protein Kinase C 동종효소 Alpha 단백발현의 특성)

  • Nam, Su Bong;Bae, Yong Chan;Park, Suk Young;Choi, Soo Jong
    • Archives of Plastic Surgery
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    • v.34 no.6
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    • pp.679-684
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    • 2007
  • Purpose: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of $PKC{\alpha}$, and ${\mu}$ was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of $PKC{\alpha}$ on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. $PKC{\mu}$ will be investigated in further study. Methods: The study was conducted with the cultured HUVECs group(control) and the $0.75{\times}10^{-9}M$ DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of $PKC{\alpha}$ was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular $PKC{\alpha}$ particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. Results: The Western blot analysis showed increased $PKC{\alpha}$ expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. Conclusion: We believe that $PKC{\alpha}$ plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

Properties of Chorismate Mutase from intrasporangium sp. (Intrasporangium속 방선균의 Chorismate Mutase 성질)

  • 조원대;신광순;최용진;양한철
    • Microbiology and Biotechnology Letters
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    • v.16 no.4
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    • pp.310-315
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    • 1988
  • Two isoenzymes of chorismate mutase(E.C.5.4.99.5) designated as chorismate mutase I(CM I) and chorismate mutase II(CM II), were detected and partially purified from a sp. of intrasporangium isolated from soil. CM I and CM II had pH optima of pH 6.5 and 8.0, respectively and showed the same temperature optimum of 45$^{\circ}C$. The activation energy of the enzymatic reaction was estimated to be 14.7kcal/ mole with CM I and 10.8kcal/mole with CM II. The affinity of isoenzyme CM I for substrate(Km= 1.35mM) was almost the same level as that of CM II(Km = 1.22mM). Both isoenzymes were stable at pH values ranged from pH 6.5 to 9.0, but rapidly denaturated at temperatures above 45$^{\circ}C$. CM II was activated about 7$^{\circ}C$ of its activity by $Ba^{++}$ or $Mg^{++}$ while CM I was slightly inhibited by the same metal ions. Thiol compounds were found not to be necessary for stability of the two enzymes but Co$^{++}$ and EDTA had a little stabilizing effect on CM II only. p-Chloromercuribenzoate strongly inactivated the activities of both enzymes but the reducing agents such as dithiothreitol and L-cysteine protected them against the pCMB inhibition.

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Characterization of Thermolabile Pectinesterase and Thermostable Pectinesterase Separated from Valencia Orange (Valencia 오렌지로부터 분리 정제한 비내열성 및 내열성 Pectinesterase의 성질)

  • Hou, Won-Nyoung;M.R., Marshall
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.666-672
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    • 1995
  • This study was carried out to characterize thermolabile pectinesterase (TLPE) and thermostable pectinesterase (TSPE) separated from crude PE of Valencia orange in order to investigate the preventive measures of cloudy juice clarification. The TLPE was observed to be mixture of several isoenzymes with the same molecular weight of 36 KD (37.5 KD) but different isoelectric point of pH 8.4, 8.7, 8.9, 9.8 and ${\geq}10$ which were unstable at $70^{\circ}C$, and the TSPE also was found to be mixture of two or three isoenzymes with the same molecular weight of 53 KD (50 KD) but different isoelectric point of pH 8.7, 9.2 and ${\geq}10$ which had slightly different stability from one another at $70^{\circ}C$. The TLPE and the TSPE had the optimum reaction pH of 7.0 and $7.0{\sim}8.5,\;appK_{M}$ of 1.1 and 1.7 mg/ml, appVmax of 0.53 and $1.01\;{\mu}mol/min/{\mu}g$, and the turnover number of 19.000 and 54,000 mol/mol/min toward Kodak pectin, respectively. The TSPE had higher storage stabiblity and cloud loss effect on orange juice than the TLPE. Above all, the crude PE was most effective on orange juice cloud loss among the PEs used.

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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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Studies on the Serum Total Creatine Phosphokinase (CPK) Activities and CPK Isosenzymes Fractions of Chicken (닭의 혈청 Creatine phosphokinase (CPK) 총 활성 및 CPK isoenzymes분획에 관한 연구)

  • Kim, Bong-Sik;Hurh, In;Kim, Jong-Hyung;Jang, Myong-Whan;Kim, Won-Sun;Kim, Duk-Whan
    • Korean Journal of Veterinary Service
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    • v.16 no.2
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    • pp.120-126
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    • 1993
  • Clinically healthy 66 female Maniker breed chicken(22 of 2-week-old : group A, 23 of 8-week-old: group B and 21 of 27-week-old: group C) and 66 female Manina breed chicken (21 of 2-week-old : group D, 22 of 8-week-old : group E and 23 of 27-week-old : group F) were examined to establish physiological basic data on serum total Creatine phosphokinase (CPK) activities and CPK isoenzymes fractions. The results obtained were summarized as follows; 1. Serum total CPK activities were $1,088{\pm}254.0\;IU\;/{\ell},$ $1,454{\pm}337.2\;IU\;/{\ell}$ and $1,440{\pm}526.3\;IU\;/{\ell}$ in gorup A, group B and group C of Maniker breed, respectively. Group B and group C showed higher values than that of group A (P<0.01) meaning high values nth aging. 2. Serum total CPK activities were $1,676{\pm}420.5\;IU\;/ {\ell},$ $1,007{\pm}283.1\;IU\;/{\ell}$ and $862{\pm}294.5\;IU/{\ell}$ in group D, group E and group F of Manina breed, respectively. Group D showed the highest value among groups (P<0.01) and Manina breed showed the low values of serum total CPK activities with aging. 3. Manina breed at 2-week-old and Maniker breed at 8-week-old and 27-week-old showed significant high values of total serum CPK activities in breed differance (P<0.01) 4. In the pattern of serum CPK isoenzyme fractions, group A and group B were high with decreasing order of $CK_3>CK_2>CK_1$ and group C was high with decreasing order of $CK_2> CK_3>CK_1$ Groihp D, E and F showed the same pattern with decreasing order of $CK_2>CK_3> CK_1.$ 5. Significance of CPK isoenzymes fractions in breed differance was only found at 8-week-old. $CK_1\;and \;CK_3$ in Maniker breed (P<0.05), and $CK_2$ in Manina breed were higher than that of the other breed (P < 0.01).

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Mechanistic Insights into the Chemopreventive Action of Phenethy1 Isothiocyanate against Ν-Nitrosobis(2-Oxopropyl) Amine-Induced Carcinogenesis

  • Lee, In-Seon
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.227-229
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    • 1996
  • The effects of phenthyl isothiocyanate(PEIFTC) on xenobiotic metabolizing enzymes and cell kinetics in the target organs for Ν-nirtosobis(2-oxopropyl) amine(BOP)-tumorigenicity were investigated in female Syrian golden hamsters in order to gain the mechanistic insigths into the chemopreventive action of PEITS against BOP-initiated lung and pancreatic carcinogenesis in hamsters. Hamsters were given BOP subcuteneo-usly(s.c.) and/or PEITC by gavage 2h prior to the BOP treatment. Eight and 24h after the PEITC administration, animals were sacrificed for analyzing P450 isoenzymes, glutathine(GSH), glutathione S-transferase(GST) and cell kinetics. The PEITC pretreatment significantly reduced the hepatic P450 isoenzume levels such as CYP2B1 and DYP1A1 which were significantly increased by the BOP treatment. However, PEITC did not affect the CYP levels in the pancreas and lung. Interestingly, the PEITC pretreatment rather lowered the heparic GST and GSH levels, regradless of BOP administration. Proliferating cell nuclear antigen(PCNA)- labeling indices were dose dependently decreased by PEITC in the pancreas acini and ducts, bronchioles, and renal tubules in which the cell replication was significantly affected by BOP. These results thus suggest that PEITC exerts the chemopreventive effects in hamsters by influencing xenobiotic matabolizing phase I enzymes in the liver and regulating cell kinetics in the target organs.

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