• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.108-108
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• 2002

포유동물의 암컷 생식기관에 존재하는 다양한 종류의 matrix metallo-proteinase (MMP)는 난소와 자궁의 구성성분의 주기적인 변화를 조절하며 이중 난소의 MMP는 난포의 성장과 배란 그리고 퇴화 동안 조직재구성에 매우 중요한 역할을 한다고 알려져 있다. 본 실험에서는 근래에 새로 발견된 사람의 난포액에 존재하는 분자량 약 110kDa인 MMP-2 isoform GA110을 동정하고 자 하였다. 난포액으로부터 GA110 단백질을 분리하기 위하여 난포액에 5mM ethylenediaminetetraacetic acid(EDTA)를 처리한 후 DEAE Sepharose Fast Flow를 이용한 chromatography를 시행하였다. 그 결과, 난포액 단백질들은 0.2M NaCl 의 분획에서 GA110 활성을 나타내었고 anti-human MMP-2 antibody에 대한 면역반응도 뚜렷이 나타났다. DEAE Sepharose Fast Flow에서 얻은 분획 중 GA110의 활성과 면역반응을 모두 나타내는 분획만을 모아 Gelatin Sepharose 4B affinity chromatography로 다시 분리하였다 분리한 결과 resin에 흡착된 단백질 (eluate) 분획들에서 매우 뚜렷한 GA110 gelatinase 활성을 나타내었으며 면역반응 또한 관찰되었다. 이 분획들의 단백질을 농축한 후 zymography를 시행하여 나타난 GA110 단백질 band를 잘라 내었으며 이로부터 단백질을 electroelution하여 농축한 후 reducing agent인 2-mercaptoethanol를 처리하였다. 이를 전기영동 후 MMP-2 (propeptide region) antibody를 사용하여 immunoblotting 한 결과 70-72kDa의 단백질만이 면역반응을 나타내었다. 마지막으로 위와 같이 준비된 70-72kDa 단백질의 아미노산 서열을 Edman degradation 방법으로 분석하였다. 그 결과 이 단백질의 N 말단의 10개의 아미노산 배열 순서가 알려진 사람의 proMMP-2의 전체 배열순서 중 propetide domain의 N 말단에서부터 다섯 번째에서 시작하여 10개의 아미노산의 서열과 정확하게 일치하였다. 위 결과들로 미루어 사람의 난포액에 존재하는 MMP-2의 새로운 isoform인 GA110은 70-72kDa의 ProMMP-2가 disulfide bond를 통해 homodimer 구조를 이루고 있는 것으로 여겨진다.

• Korean Journal of Animal Reproduction
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• v.16 no.3
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• pp.209-215
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• 1992

These studies were carried out to overcome 2-cell block and in vitro development to blastocysts in vitro fertilization of mouse embryos. The unfertilized ova were obtained by superovulation in ICR mice of 4 to 6 weeks old. Tyrode's 280 solution was used as basal media, and the pH range of media examined was designed from 6.5 to 7.5 with 0.2 interval and the range of osmolality from 250 to 370 mOsm with 20 interval, and the period of sperm preincubation examined was 30, 60, 120, and 180 minutes. The ova developed to 2-cell embryos after 26hrs of incubation with preincubated sperm were evaluatated as in vitro fertilized ones. The results obtained were summarized as follows: 1. The optimal ranges of pH and osmolality of culture media and of sperm preincubation time for in vitro development of in vitro fertilized ova to blastocyst were pH 7.1 to 7.3, 250 to 350 mosmol and 60 to 180 min, respectively. 2. With the media of pH 7.1, 310 mOsm and sperm preincubation period of 120min in another experiment of large sample size, the in vitro fertilized ova was found 66.5% and the in vitro development of in vitro fertilized ova to blastocyst was found 35.8%. From the above results it was concluded that the optimal conditions of pH and osmolality of the media for mouse IVF and embryo culture, and the period of sperm preincubation might be 7.1, 310 mOam and 120min, respectively.

• The Korean Journal of Mycology
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• v.32 no.2
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• pp.112-118
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• 2004

Laccase produced by Trametes hirsuta S1 isolated from Korea was partially purified and characterized using ultrafiltration, anion exchange chromatography and affinity chromatography. The laccase was produced as the predominant extracellular enzyme during primary metabolism. Neither lignin peroxidase nor veratryl alcohol oxidase (VAO) were detected in the culture fluid. Addition of 2,5-xylidine enhanced 4-fold laccase production. Purified laccase was a single polypeptide having a molecular mass of approximately 66 kDa, as determined by SDS-polyacrylamide gel electrophoresis, and carbohydrate content of 12%. $K_{m}\;and\;V_{max}$ values for laccase with ABTS [2,2-azinobis (3-ethylbenzthiazoline 6-sulfonic acid)] as a substrate (Lineweaver-Burk plot) was determined to $51.2\;{\mu}M\;and\;56.8\;{\mu}mole$, respectively. The optimal pH for laccase activity was found to be 3.0. The enzyme was very stable for 1 hour at $50^{\circ}C$. Half-life ($t_{1/2}$) of the enzyme was about 20 min at $70^{\circ}C$. Spectroscopic analysis of purified enzyme indicated that the enzyme was typical of copper-containing protein. Substrate specificity and inhibitor studies for laccase also indicated to be a typical fungal laccase. The N-terminal amino acid sequence of the T. hirsuta S1 laccase showed 100% of homology to those of laccase from C. hirsutus.

• Applied Biological Chemistry
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• v.12
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• pp.69-74
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• 1969

This glucose isomerizing enzyme, which Actinomyces sp. (strain, K-17) produced, was inhibited by citrate, oxalate, ethylene diamine tetraacetic acid and cysteine on the enzyme reaction. This enzyme isomerized xylose to ketose as well as glucose. The Michaelis constant of this enzyme was $7.2X\;10^{-1}M.$. on D-glucose. The enzyme activity of intact cells which were harvested in the none xylose containing medium was very weak. If these intact cells of low activity were treated in the buffered xylose solution, its activity was considerably increased. After fifteen hours at $32^{\circ}C$. on xylose treatment, the enzyme activity was increased to equilibrium and the treating condition was proper at neutral pH and in aerobic state. The adequate concentration of xylose on the treatment was about 0.5 percent.

• Korean Journal of Environmental Agriculture
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• v.23 no.2
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• pp.92-98
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• 2004

The adsorption and desorption of Pb, Cd, Co, Zn, Cr, Co, Ni, and Mo on the waste Undaria sp. were studied. Except for Pb. the mono adsorption rate for all heavy metals were lower than that of the heavy metals mixed. However, the adsorption capacity of the heavy metals by 1g of biosorption, in mixed heavy metals increased According to FT-IR analysis of the biosorbent after heavy metal biosorption, the replacement of the functional group by the heavy metals ions could be confirmed and the inverted peaks became larger after heavy metals adsorption. The adsorption equilibrium of heavy metals was reached in about 1 hour. The equilibrium parameters were determined based on Langmuir and Freundlich isotherms. The affinity of metals on the biosorbent decreased in the following order: Pb>Cu>Cr>Cd>Co. The desorption rate decreased in the following sequence: NTA>$H_2SO_4$>HCl>EDTA. The desorption rate of heavy metals by NTA increased with increase in the concentration from 0.1 to 0.3% but the desorption rate became constant beyond 0.3%. Therefore, it represented that desorption rate of heavy metals was suitable under optimized condition ($30^{\circ}C$, pH 2 and 0.3% NTA solution) and was fast with 80% or more the uptake occurring within 10 min of contact time.

• Korean Journal of Agricultural Science
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• v.41 no.1
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• pp.9-16
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• 2014

This experiment was conducted to examine the effects of a short-term treatment of high pressure $CO_2$ on shelf-life of strawberry fruit. A short-term treatment (12 hr) of 100% $CO_2$ resulted in the increase of fruit firmness up to 71.9% compared to that at harvest. The firmness of $CO_2$ treated fruit remained a significantly higher than that of control (air) up to 15 days. The alteration of pectic polymers was observed by $CO_2$ treatment such as an increase of EDTA soluble pectins and decrease of water soluble ones. The $CO_2$ treatment resulted in the increase of total amount of wall bound calcium. Pectate lyase activity, an important agent of strawberry fruit softening, was also significantly reduced by $CO_2$ treatment. Contents of soluble solids and acids of $CO_2$ treated fruit were higher than those of control fruit. Short-term treatment of high pressure $CO_2$ affected shelf-life through firmness increase whereas the visual quality and decay incidence of strawberry fruit were not affected.

• Korean Journal of Environmental Agriculture
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• v.23 no.1
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• pp.28-33
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• 2004

This study was conducted to how the effect of lime and humic acid on cadmium availability and ie uptake by plant grown in contaminated paddy soils with heavy metal. The treatment levels of lime were 2.5 and 5.0 ton/ha and that of humic acid were 1 and 2%. The contents of 0.1N HCl extractable Cd were reduced with lime and humic acid and were negatively correlated with CEC as well as soil pH. The sequential extraction procedure was used to fractionate the heavy metals in soils into the designated from exchangeable (0.5 M $KNO_3$) water soluble ($H_2O$), organically bound (0.5 M NaOH), carbonate (0.05 M $Na_{2-}$ EDTA) and sulfide/residual (4 M $HNO_3$). In soil amended with 2.5 ton/ha lime and 1% humic acia che- mical forms of Cd at tillering stage were predominant exchangeable + water soluble extractable Cd, whereas that at harvesting stage were predominant carbonate + sulfide/residual extractable Cd. The exchangeable forms of Cd in soil with lime and humic acid were negatively correlated with soil pH during the harvesting period. Total absorbed Cd of paddy rice tended to occur in the order of root > stem > leaf > brown rice. Cd contents of brown rice with lime and humic acid treatment were 0.09 and 0.08 mg/kg, respectively. That were lower than control, 0.20 mg/kg. It could be that treatment of lime and humic acid in polluted soil by heavy metals would reduce the uptake of heavy metals by piano and be a temporary method of reclamation at the highly heavy Metal contaminated soils.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.8 no.2
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• pp.90-100
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• 1975

The estimation of the pre-processing condition of oyster is of great importance for distributors and processors. This study was attempted to establish the basic data for evaluating the processing suitability of oyster, which is the most important shellfish for domestic use and export. The data were analysed by measuring the condition index, chemical composition and heavy metal content of oysters. In order to eliminate the manual work that has to be done on a tightly closed oyster shell and avoid shrinkage in the oyster meat which is attendant on the steaming process, chemical means to open oyster were examined. finding the method of pretreatment of polyphosphate for frozen oysters were attempted to improve the product quality. The prevention of undesirable color change of the canned oyster meat is another problem to solve. The important results are as follows : 1. The ratio of meat volume and meat weight to the holding capacity by shells may be useful as an index to measure the condition index of oysters. 2. As a whole, monthly changes of moisture and fat content in oysters were reversely correlated. Protein content slightly decreased from April and rapidly decreased in July, and again rapidly increased in August but from September to November decreased slightly. In April, the content of glycogen was 4 percent. From this period to September, glycogen was rapidly decreased. From July to September, it was only 0. 7 to 1 percent but increased from October. There were little seasonal changes in pH value. The pH value of oyster meat was 6.0 to 6.2. The crude ash content was slightly decreased from June to August. 3. The range of monthly change of heavy metal content are as follows: Total mercury was 0 to 0.019 ppm, cadmium was 0.026 to 0.053 ppm, copper was 0.111 to 0.594 ppm, and lead_was 0.061 to 0.581 ppm. 4. By the results of condition index, chemical composition and heavy metal content of oysters, the suitable harvest season as raw materials for processing was the end of December to the end of May of next year. 5. The pretreatment of 10 percent polyphosphate in 5 percent salt solution of oyster meat appeared effective to reduce thawing drip during cold storage. 6. The pretreatment of $Na_2EDTA$ and BHA did not show the color prevention effect to the canned oyster meat during storage. 7. Magnesium chloride affected to open the valves of oysters.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.199-206
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• 2012

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol ($39.85%{\pm}11.41$) than in 5% glycerol treatment ($18.08%{\pm}1.61$). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol ($34.12%{\pm}11.02$) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

• Journal of Periodontal and Implant Science
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• v.24 no.3
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• pp.581-596
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• 1994

This in vitro study was undertaken to obtain optimal tetracycline concentration that aids proliferation and spreading of human periodontal ligament cells, for clinical application in root surfaces of periodontally diseased teeth. Periodontal ligament cells used in this study were obtained from explants of periodontal ligament of 1st premolar teeth which were extracted for the purpose of orthodontic treatment. The cells were cultured in Dulbecco's Modified Eagle Medium(DMEM) supplemented with 100 U/ml penicillin, $100\;{\mu}g/ml$ streptomycin and 10% FBS at $37^{\circ}C$, 100% humidity, 5% $CO_2-95%$ air. Cells were used between the third to 4th passage. After root planing of periodontally extracted teeth, the root slabs were cut with carborundum disk. In the cell proliferation experiment, experimental groups were root planing only group, immersed groups in 25, 50, 75, 100, 150mg/ml aqueous solution of Tetracycline HCl followed by a vigorous rinse in PBS. Human PDL cells at concentration of $1{\times}10^5\;cells/ml$ were seeded in each culture well which contained root slabs and incubated for 6 hours. Then, all of the root slabs were moved into new 24 culture well and incubated 24, 48 and 72 hours. The cell counting was done by inverted phase contrast microscope after trypsinization. The following results were obtained. The cell number was increased in order root planing only group, 25, 150, 50, 75, 100mg/ml of Tetracycline HCl treated group in 24, 48 and 72 hours. The maximal cell number was obtained when the root slabs were immersed in solution with 100mg/ml of Tetracycline HCl. There were statistically significant between the root planing only group and 75, 100 mg/ml of Tetracycline HCl treated group in 24 hours, between the root planing only group and 100mg/ml of Tetracycline HCl treated group in 48 hours, between the root planing only group and 50, 75, 100mg/ml of Tetracycline HCl treated group, between 25 and 100mg/ml of Tetracycline HCl treated group in 72 hours(p<0.05). In the cell spreading experiment, after 30 minutes of incubated, in the root planing only group, the cells were generally round in shape. The cell surface was mostly covered with blebs. The cells started to attach to root surface by cytoplasmic extension in 50, 100mg/ml of Tetracycline HCl treated groups, more numerous cells attached to root surface than root planing only group. Many orifices of dentinal tubule were exposed, cells showed radially spreaded cytoplasm and unspreaded central region of the cell was covered with blebs. After 6 hours of incubation, in the root planing only group, cells showed radially spreaded cytoplasm and were attached flat appearance. In 50, 100mg/ml of Tetracycline HCl treated groups, cellular margin was concaved and cytoplasm showed elongated appearance with polarity. After 24 hours of incubation, in the root planing group, cells showed characteristic polarity. In 50, 100mg/ml of Tetracycline HCl treated groups, cells showed more elongated and spindle - like appearance.