• Journal of Microbiology
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• v.37 no.1
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• pp.14-20
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• 1999

Two extracellular proteases, EP I and EP II, from cells of Oligotropha carboxydovorans (formerly Pseudomonas carboxydovorans) DSM 1227 grown in nutrient broth were purified to greater than 95% homogeneity in five steps using azocasein as a substrate. The final specific activities of EPs I and II were 214.9 and 667.4 units per mg of protein. The molecular weights of native EPs I and II were determined to be 23,000. Sodium dodecyl sulfate-gel electrophoresis revealed the two enzymes to be monomers. The enzymes were found to be serine-type proteases. The activity of EP I was stimulated by Ca2+, Mg2+, and Ba2+, but that of EP II was not. The enzymes were completely inhibited by Fe2+, Hg2+, Co2+, Zn2+, and Cd2+. EDTA and EGTA exhibited a strong inhibitory effect on EP I. The optimal pH for the two enzymes was pH 9.0. The optimal temperatures for EP I and II were 60 and 50$^{\circ}C$, respectively. The enzymes were stable under alkaline conditions. The thermal stability of EP I was higher than that of EP II. Cell-free extracts did not inhibit the purified enzymes. The enzymes were active on casein, azocasein, azocoll, and carbon monoxide dehydrogenase, but weakly active with bovine serum albumin.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.551-563
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• 1996

This study investigated the properties of primary cultured proximal tubule cells in hormonally defined(insulin, transferrin, and hydrocortisone), serum-free medium or 10% serum-supplemented medium. The growth rate of the primary cultured proximal tubule cells was lower in the hormonally defined, serum-free medium than in the 10% serum- supplemented medium(p < 0.05), while the activities of brush border marker enzymes, alkaline phosphatase(AP), leucine aminopeptidase(LAP), and y-glutamyl transpeptidase(${\gamma}$-GTP) were increased(p < 0.05). The activities of these enzymes, however, decreased with the lapse of incubation time to 50-70% after 6 days culture compared to those of the freshly-prepared proximal tubules. The enzymatic activities of the primary cultured proximal tubul cells on 6, 9, 12, and 15 days of culture were significantly increased in the hormonally defined, serum-free medium compared to the 10% serum-supplemented medium(p < 0.05). The functional differentiation of the primary culture was examined by observing multicellular domes of the confluent monolayer, which is indicative of transepithelial solute transport. The dome formation by the proximal tubule cultures occurred at a higher frequency in the hormonally defined, serum-free medium than in the 10% serum-supplemented medium(p < 0.05). Upon electron microscopic examination, an increased density of the brush border was observed in the hormonally defined, serum-free medium compared to the cells grown in 10% serum-supplemented medium. The activities of $Na^+$glucose cotransporter($^{14}C$-a-MG uptake), $Na^+$phosphate cotransportere($^{32}P$ uptake) and $Na^+$ transporter($^{22}Na^+$ uptake) in the brush border membrane, and of $Na^+/K^+$-ATPase($^{86}Rb$ uptake) in the basolateral membrane were significantly stimulated in the hormonally defined, serum-free medium than in 10% serum-supplemented medium(p < 0.05). In conclusion, the primary cultured proximal tubule cells grown in the hormonally defined, serum-free medium demonstrated a slower growth rate, but the functions of cell were enhanced.

• Toxicological Research
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• v.18 no.2
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• pp.159-165
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• 2002

Recently, we reported (korean J. Biomed. Lab. Sci., 6(4): 245-251, 2000) that cyclohexane (l.56 g/kg of body wt., i.p.) administration led to lung injury in rats. However the detailed mechanism remain to be elucidated. This study was designed to clarify the mechanism of lung damage induced by cyclohexane in rats. First, lung damage was assessed by quantifying bronchoalveolar lavage fluid (BAL) protein content as well us by histopathological examination. Second, activities of serum xanthine oxidase (XO), pulmonary XO and oxygen free radical scavenging enzymes. XO tope conversion (O/D + O, %) ratio and content of reduced glutathione (GSH) were determined. In the histopathological findings, the vasodilation, local edema and hemorrhage were demonstrated in alveoli of lung. And vascular lumens filled with lipid droplets, increased macrophages in luminal margin and increased fibroblast-like interstitial cells in interstitial space were observed in electron micrographs. The introperitoneal treatment of cyclohexane dramatically increased BAL protein by 21-fold compared with control. Cyclohexane administration to rats led to a significant rise of serum and pulmonary XO activities and O/D + O ratio by 47%,30% and 24%, respectively, compared witれ control. Furthermore, activities of pulmonary oxygen free radical scavenging enzymes such as superoxide dismutase, glutathione peroxidase and glutathione S-transferase, and GSH content were not found to be statistically different between control and cyclohexane-treated rats. These results indicate that intraperitoneal injection of cyclohexane to rats may induce the lipid embolism in pulmonary blood vessel and lead to the hypoxia with the ensuing of oxygen free radical generation, and which may be responsible for the pulmonary injury.

• The Journal of Internal Korean Medicine
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• v.40 no.6
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• pp.1219-1224
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• 2019

Objective: To investigate the possibility of hepatotoxicity by supplemental foods or folk herbs such as Salvia Plebeia R. Br. Methods: A hospitalized male patient with alcoholic liver disease and electrolyte imbalance had recovered, and then followed by rapid hepatic serum enzymes after taking S. Plebeia. This study monitored the clinical outcome and biochemical parameters. Result: A 58-year male had drunk frequently, which led to alcoholic steatohepatitis and hospitalization. Two weeks after his discharge from hospital, he felt nausea, dizziness, and mild difficulties in speech and walking, resulting in re-hospitalization at the Korean Medical Hospital. The symptoms disappeared on correction of the electrolyte imbalance suspected to have been caused by severe sweating while working in the outdoors, and the patient was discharged. During treatment and monitoring of his health as an outpatient, the serum hepatic enzyme rapidly elevated approximately 10-fold in hepatic enzymes; the enzyme levels fluctuated according to whether or not he was taking the boiled water of S. Plebeia. The RUCAM score was 12, which met the criteria for toxic hepatitis by S. Plebeia. His general condition and abnormal hepatic enzymes recovered with cessation of S. Plebeia and administration of Chungganplus syrup (CGX). Conclusion: This study reports the hepatotoxic risk of Salvia Plebeia, which is commonly used as a folk remedy in Korea.

• Journal of the Korean Wood Science and Technology
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• v.21 no.3
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• pp.87-93
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• 1993

This study was undertaken to investigate the characteristics and the productivities of lignin olytic enzymes: laccase (Lac) and Mn-dependent peroxidase (MnP) from Coriolus versicolor and Lentinus edodes respectively. Enzymes were isolated from cultural filterates and purified according to the standard methods. These enzymes showed one band in SDS-PAGE and their molecular weights were found 62,000 and 45,000 dalton respectively. Polyclonal antibodies against Lac and MnP were raised against mouse. In the ELISA (enzyme-linked immunosorbent assay), Lac and MnP-antiserum produced a strong positive reaction with Lac and MnP antigen($A_{405}$=2.50 and 3.53 respectively). The sera to negative (S/N) ratio was determined by the dividing the mean absorbance of antibodies by the corresponding diluted samples from normal mouse serum. The sera produced showed 2 times more positive reaction in S/N ratio than negative sera.

• Journal of Life Science
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• v.7 no.4
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• pp.384-391
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• 1997

The purpose of this study was to investigate effects of swimming training and cholesterol diet on the activation of hepatic antioxidant enzymes and serum lipid in Sprague Dawley rats(24 weeks of age). They were divided into five groups which were made up of normal-diet detraining group(C), 2% cholesterol-diet detraining group(CC), 2% cholesterol-diet swimming training groups which were classified according to their training time(CSA: 12min, CSB:8min, CSC:4min). They were given normal diet for the first 6weeks and then, separated normal-dietary and 2% cholesterol-dietary for 14 weeks. During these periods, 10 weeks’ swimming training was performed after 4 weeks later. And then we analyzed blood and liver by decapitating those rats. Swimming training showed a tendency to increase the activation of GSH-peroxidase, Nonprotein-SH and malondialdehyde, and decrease total- cholesterol, LSL-C/HDL-C and VLDL significantly. Whereas, cholesterol diet which has no training showed decrease the activation of hepatic antioxodant enzymes, and increase total-cholesterol and LDL-C/HDL-C absolutely. These results suggest that swimming training should stimulate the activation of hepatic antioxidant enzymes and decrease total-cholestrol even if they had cholesterol diet.

• Journal of Chest Surgery
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• v.22 no.4
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• pp.562-572
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• 1989

The changes of serum glutamic oxaloacetic transaminase [SGOT], serum glutamic pyruvic transaminase [SGPT], serum lactic dehydrogenase [LDH] and serum creatine phosphokinase [CPK] were examined during and after the open heart surgery. In the total of 52 patients with heart diseases including 40 cases of congenital heart anomalies and 12 cases of acquired valvular heart diseases who undergone open heart surgery under cardiopulmonary bypass. The results obtained are as follows: 1. The average value of SGOT before surgery was 30.27 [ 18:86 units. The enzyme was reached to the maximum of 139.88 [ 89.43 units on the 1st day after the operation [p< 0.05], the enzyme activity was gradually decreased from the 3rd day after the operation, returned to the normal range on the 7th day after the operation. 2. The average value of SGPT before surgery was 14.36 [ 7.45 units. The enzyme was reached to the maximum of 34.67 [ 27.64 units on the 2nd day after the operation, but it was valueless statistically, the enzyme activity was gradually decreased from the 3rd day after the operation, returned to the normal range on the 5th day after the operation. 3. The average value of LDH before surgery was 263.07 * 86.66 units. The enzyme was reached to the maximum of 662.29 * 303.60 units on the 2nd day after the operation [p < 0.05], the enzyme activity was gradually decreased from the 5th day after the operation, returned to the normal range on the 7th day after the operation. 4. The average value of CPK before surgery was 141.35 * 43.44 units. The enzyme was reached to the maximum of 377.42 [ 222.02 units on the 1st day after the operation [p < 0.05], the enzyme activity was gradually decreased from the 5th day after the operation, returned to the normal range on the 7th day after the operation. 5. In the relationships of the serum enzymes and duration of the extracorporeal circulation, the values on the group over 90 minute of the extracorporeal circulation were increased than on the group below 90 minute of the extracorporeal circulation, but it was valueless statistically.

• BMB Reports
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• v.31 no.5
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• pp.519-523
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• 1998

We have previously reported that anti-glucose-6-phosphate dehydrogenase (G6PD) serum raised against native G6PD (nG6PD) enzyme recognized nG6PD antigen poorly in competitive enzyme-linked immunosorbent assay (ELISA) (Kim, 1997). In the present study, we investigated whether anti-G6PD serum raised against nG6PD can react with denatured G6PD effectively in competitive ELISA. We used partially active G6PD (paG6PD) by repeated freeze-thawing or SDS-denatured G6PD (SDS-G6PD) as both immobilized and soluble antigens, and anti-G6PD serum raised against nG6PD for competitive ELISA. The polystyrene cuvettes coated with either paG6PD or SDS-G6PD were challenged with a mixture of a limiting amount of anti-G6PD serum and various doses of paG6PD or SDS-G6PD as competitors, followed by incubation with alkaline phosphatase-anti-IgG conjugate. The competitive ELISA with paG6PD or SDS-G6PD antigen exhibited the sigmoidal dose-response curve characteristic of competition immunoassays. Furthermore, Triton-denatured G6PD (Triton-G6PD) was used in competitive ELISA. The paG6PD, SDS-G6PD, or Triton-G6PD used as competitors increased the inhibition of antibody binding to immobilized either of nG6PD or denatured G6PD compared with nG6PD competitor. The inhibition by denatured G6PD competitors was more pronounced at high competitor concentrations than at low counterparts. We conclude that anti-G6PD serum raised against nG6PD can effectively react with denatured G6PD in competitive ELISA and that our anti-G6PD serum recognizes denatured enzymes better than active enzymes.

• YAKHAK HOEJI
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• v.12 no.1_2
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• pp.1-15
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• 1968

The levels of activities of amylase, lipase and trypsin in both the pancreatic tissue and serum of 18 species of vertebrate animals were measured and enzymologically compared with each other. 1) The value of amylase in the pancreas of experimental mammalia has been found decreasing in the order pig, rat, dog, cat, rabbit and cow; that of pancreatic lipase has been found decreasing in the order of pig, dog, cat, rat, rabbit and cow; and that of trypsin has been found decreasing in the order of pig, cow, dog, rat, rabbit. Thus the value of all the above three kinds of enzymes were observed highest in pig, but in cow amylase and lipase were observed lowest while trypsin were observed considerably high. 2) In view of diets, the comparatively high values of pancreatic enzyme were observed in the ommivorous animals such as pig, rat, dog, while the values observed low in the herbivorous animals, such as cow and rabbit. 3) In the bovine, the values were observed moderately high except lipase which were found comparatively low. 4) In the Reptilia and Amphibia such a mud turttle and frog, the values were shown in similar measure with each other, that is, the pancreatic amylase and trypsin were observed considerably high while the lipase was found low. 5) In the species of Reptilia such as a viper and snake, the activities of pancreatic enzymes were not detected. But in the tissue of liver, stomach, activities of the enzymes were found considerably high. Lacertilia animals such as lizard the values of pancreatic enzymes were little observed. 6) In the fish in which the pancreatic tissue is scattered in the liver, the pancreatic enzymes were found in the liver tissue considerably higher than in the other tissues but lower than in the warm-blooded animals, especially the lipase was lower. 7) In generally the values of serum amylase and lipase were observed higher than those of man; and even in the cold-blooded animals in which the values of pancreatic enzymes were shown low or none, the values were also observed high. 8) The above three kinds of pancreatic enzyme values of those experimental animals have shown a tendency of higher degree in higher taxa than in lower taxa according to taxonomical order. 9) In view of tissue, the pancreatic cell was observed large in the mammalian animals such as rat and pig and cytoplasm was also abundantly contained in the acinous cell; and the bovine and the snake haave the pancreatic cells of the similar rosette form the comparatively large acinous cells of long rhombic form in the comparatively large acinous cells of long rhombic form in which the spindle shaped neucleus and the abundant cytoplasm were contained. In the fish the pancreatic cell were found scattered in the liver in which the very large pancreatic islet were found.

• Environmental Analysis Health and Toxicology
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• v.17 no.4
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• pp.307-314
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• 2002

Effects of ($\pm$)-camphor on liver cytochrome P450 enzymes were investigated in male ICR mice. Mice were treated intraperitoneally with 0, 200, 400 and 800 mg/kg of ($\pm$)-camphor in corn oil for 3 consecutive days. Twenty four hr after the final treatment, the animals were subjected to necropsy. The activities of serum aspartate aminotransferase and serum alanine aminotransferase were slightly changed by the treatment with ($\pm$)-camphor at the doses used. Administration of ($\pm$)-camphor to mice significantly induced the hepatic activities of pentoxyresorufin O-depentylase and benzyloxyresorufin O-debenzylase and weakly induced ethoxyresorufin O-deethylase in dose-dependent manners. The present results suggested that ($\pm$)-camphor might act as a relatively specific inducer of hepatic cytochrome P450 2B in male ICR mice.