• Journal of Embryo Transfer
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• v.10 no.2
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• pp.145-151
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• 1995

초기배의 성판정은 대상가축의 성을 선발하는 수단으로써 가축의 육종 및 번식에 있어 가치가 매우 높다. 체세포, 체외수정 또는 처녀발생 초기배의 성을 결정하기 위해 capillary polymerase chain reaction (PCR)을 이용하였으며 성판정에 이용되는 상실배 또는 배반포는 체외수정과 그 후의 난관상피세포와의 공배양에 의해 생산되었다. 초기배의 genomic DNA는 0.2$\mu$g/$\mu$L proteinase K를 함유하고 있는 PCR lysis buffer에 하나의 초기배를 부유하게 50˚C 에서 1시간동안 배양한 후 95˚C에서 10분간 효소를 비활성화시킴으로써 준비되었다. 웅성 초기배에서는 두개의 증폭산물(소특이)만이 생산되었다. 이 기법에 의한 성판정 결과 초기배의 성비는 예상되는 1:1의 성비와 유의적인 차이가 없었다. 잔여 난구세포 또는 투명대에 결합된 정자 등으로 인한 잘못된 성판정이 종종 발생하였는데, 이는 citrate 처리후 투명대를 완전히 제거함으로써 false positive 또는 negative 결과를 극복할 수 있었다. 이상과 같은 결과는 체외생산 소 초기배의 신속하고(2시간 증폭) 정확한 성판정의 가능성과 초기배 이외의 세포로부터의 오염이 확립된 수세방법에 의해 효과적으로 배제될 수 있음을 제시하였다.

• The Korean Journal of Pesticide Science
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• v.2 no.2
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• pp.68-74
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• 1998

벼멸구의 카보후란에 대한 저항성 기작을 구명하기 위해 실내에서 카보후란으로 30세대 도태하여 얻은 저항성계통($LD_{50};\;20.3{\mu}g/g$)과 약제를 12년 동안 처리하지 않은 벼멸구 감수성 계통($LD_{50};\;0.3{\mu}g/g$)을 완충용액과 마쇄하여, 105,000g에서 2시간 원심분리하여 얻은 상등액(에스테라제층)과 침전물(P450-산화효소층)을 효소액으로 하여 $^{14}C$-카보후란을 반응시켜 계통 간 대사물 량의 차이를 조사한 바 저해제(piperonyl butoxide; 산화효소저해제, diethylmalate; 글루타치온 전이효소 저해제, iprobenfos; 에스테라제 저해제)와 보조인자 (NADPH; P-450 산화효소, 글루타치온; 글루타치온전이효소)에 상관없이 카보후란의 대사물과 그 양이 계통간 차이가 없었다. 이상의 결과로부터 저항성 벼멸구에서 일반적으로 곤충에서 생화학적 저항성 기구로 잘 알려진 가수 분해 효소의 일종인 에스테라제와 p-450 산화효소, 글루타치온 전이효소의 활성 증가가 저항성 발달에 관여하지 않음을 알 수 있었다.

• Korean Journal of Ecology and Environment
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• v.39 no.4 s.118
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• pp.435-444
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• 2006

A suite of extracellular enzyme activities involved in organic carbon decomposition were determined in three northern peatlands (a bog, a fen, and a swamp) over a 12 month period along with trace gas ($CO_2$ and $N_2O$) flux and DOC dynamics in the wetlands. The activities varied $0.008-0.066\;{\mu}mole\;g^{-1}\;min^{-1}$, $0.003-0.021\;{\mu}mole\;g^{-1}\;min^{-1}$, $0.003-0.016\;{\mu}mole\;g^{-1}\;min^{-1}$, $0.004-0.047\;{\mu}mole\;g^{-1}\;min^{-1}$, for ${\beta}-glucosidase$, cellobiohydrolase, ${\beta}-xylosidase$, and N-acetylglucosaminidase, respectively. In general, the activities were highest in the forested swamp followed by the fen and the bog. When the data from three wetlands are combined, the enzyme activities exhibited significant positive correlations with trace gas emission and available carbon. Further, the average activity of 4 enzymes explained about 20-40% of the variations of trace gas emssion and available carbon. The results indicate that enzymes related to the mineralization of organic carbon may play an important role in trace gas flux and DOC dynamics in northern peatlands.

• Microbiology and Biotechnology Letters
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• v.17 no.1
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• pp.69-73
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• 1989

Cellular autolytic enzyme was isolated from the supernatant fluid of exponentially growing cuiture of Cl. butyricum ID-113. The autolysin was partially pruified by ammonium sulfate fractionation, chromatography on DEAE-Sephadex A-50 and gel filtration through Sephadex G-200. This autolytic enzyme lysed SDS-treated cell wall fractions of Cl. butyricum ID, but not whole cells at all. Its optimum pH and temperature were 5.0 and 37$^{\circ}C$, respectively. This enzyme was relatively stable at neutral pH, but sensitive to heat treatment. Enzyme activity was not influenced by the addition of various divalent cation, but inhibited by Cu$^{++}$.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.02a
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• pp.43-45
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• 2001

• Development and Reproduction
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• v.5 no.1
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• pp.17-21
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• 2001

This study was conducted to investigate the expression pattern of Trypsin-like enzyme and the effect of a trypsin inhibitor(benzimidine) on hatching process during in-vitro culture of mouse preimplantation embryos. The Trypsin-like enzyme was identified by rhodamine-conjugated Trypsin substrate probe. The expression of trypsin-like enzyme was firstly detected at the late morula stage, and the enzyme was uniformly localized in the trophectoderm of late blastocysts. Especially, intense fluorescence was observed in the blebbing area of hatching blastocysts. Bisbenzamidine, contained in culture media, did not alter embryonic development from 4-cell stage to the expanded blastocyst but decrease the hatching rate in ImM concentration (15.8% vs 89.7%, p<0.02). In the treatment of bisbenzimidine (5mM) for 12 hours according to the embryonic stage of mouse, the hatching rate of control (83.0%) and treatment in late blastocysts (8.7%) were significantly (p<0.01) different. From these results, we suggested that the hatching enzyme having trypsin-like activity was localized from the late morula stage, and the hatching process by this enzyme was activated in the late blastocyst stage of mouse embryos.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.15 no.4
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• pp.263-270
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• 1982

In an attempt to evaluate the nutritional quality of seaweed protein, the effects of heat treatment on the in vitro digestibility and trypsin inhibitor content in seaweed were determined. In this study, the nitrogen-to-protein conversion factors were also calculated on the basis of quantitative amino acid data. The results are as follows : 1. The in vitro protein digestbilty of red seaweeds (P. teoera anc P. suborbiculata) were ranged from 78.5 to 82.2, and green seawerd (E. linza) and brown seaweeds showed value under 80 in vitro digestibility. In general, trypsin inhibitor contents in brown seaweed were higher (0.33-0.54 mg/g) than those of red seaweeds (0.26-0.39 mg/g). And it is noted that the lowest trypsin inhibitor content was shown in green seaweed (E. linza) in spite of lowest in spite digestibility (78.5). 2. The in vitro protein digestibility of sun dried laver (P. tenera) was increased with cooling time (microwave heating), but it was not significant. Hot plate cooking raised the in vitro digestibility from 81. 1 to 84.5. The influence pot cooking time on trypsin inhibitor content was inversely proportional to in vitro digestibility. 3. Computed nitrogen factor, based on amino acid content (Factor method) and Kjeldahl nitrogen content (Kjeldahl mettled), were 5.83 (H. fusiforme)- 6.52 (P. tencra) as Factor method and 5.40 (U. pinnatifida)-6.29 (P. tenera) as Kjeldahl method. Individual value for each nitrogen conversion factor differed by species, especially in brown seaweeds. The best estimate of the protein content of seaweed can be calculated, from multiplying the summed amino acid content by conversion factor (Factor method).

• Journal of Wetlands Research
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• v.11 no.1
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• pp.123-130
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• 2009

Hyporheic zone is an area where hydraulic exchanges occur between surface water and ground water. Such transient area is anticipated to facilitate diverse biogeochemical reactions by providing habitats for various microorganism. However, only a few data are available about microbial properties in hyporheic zone, which would be important in better understanding of biogeochemical reactions in whole streams. The study site is Naesung stream, located in the north Kyoung-Sang Province, of which sediment is sandy with little anthropogenic impacts. Soil samples were collected from a transect placed perpendicular to stream flow. The transect includes upland fringe area dominated by Phragmites japonica, bare soil, and soil adjacent to water. In addition, soil samples were also collected from downwelling and upwelling areas in hyporheic zone within the main channel. Soils were collected from 3 depth in each area, and water content, pH, and DOC were measured. Various microbial properties including extracellular enzyme activities ($\beta$-glucosidase, N-acetylglucosaminidase, phosphatase and arylsulfatase), and microbial community structure using T-RFLP were also determined. The results exhibited a positive correlation between water content and DOC, and between extracellular enzyme activities and DOC. Distinctive patterns were observed in soils adjacent to water and hyporheic zone compared with other soils. Overall results of study provided basic information about microbial properties of hyporheic zone, which appeared to be discernable from other locations in the stream corridor.

• Journal of Chest Surgery
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• v.31 no.5
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• pp.502-508
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• 1998

Proinflammatory cytokines such as tumor necrosis factor-$\alpha$(TNF-$\alpha$) have been implicated in myocardial and organ dysfunction associated with postperfusion syndrome. We tested the hypothesis that cytokine productions are depressed by preoperative cortiosteroid injection for cardiopulmonary bypass(CPB) and the postoperative courses will be better than without steriod pretreated cases. Cardiac surgery was performed in randomized blind fashion for 20 patients from June 1996 to September 1996. In the steroid group(n=10), corticosteroid(dexamethasone 1 mg/kg) was injected 1 hour before anesthetic induction, but in the control group(n=10), nothing was injected. Each of groups were sampled 11 times as scheduled for TNF-$\alpha$ bioassays. We have checked EKG, cardiac enzymes(CPK, LDH with isoenzyme), WBC count preoperative day, one day and three days after operation. Viatal signs were continuously monitored for three postoperaive days. In the postoperative period three patients in the control group had elevated body temperature and four patients had hypotension that required considerable intravenous fluid administration. But steroid injected patients showed normal body temperture and acceptable blood pressures without supportive treatment. CPK enzymes rose in control group higher than steroid group at postoperative 1st and 3rd day(CPK; 1122$\pm$465 vs 567$\pm$271, 864$\pm$42 vs 325$\pm$87), and CPK-MB enzymes rose in control group higher than steroid group at postoperative 1st day(106.4$\pm$115.1 vs 29.5$\pm$22.4)(P=0.02). Arterial tumor necrosis factor-$\alpha$ rose during cardiopulmonary bypass, peaking at 5 minutes before the end of aortic cross clamping(ACC-5min) in steroid group(11.9$\pm$4.7 pg/ml), and 5 minutes before the end of cardiopulmonary bypass(CPB-5min) in control group(22.3$\pm$6.8 pg/ml). The steroid pretreated patients had a shorter period of time in respirator suport time, ICU stay day, hospital admission day. We conclude that corticosteroid suppress cytokine production during and after cardiopulmonary bypass, and may improve the postoperative course through inhibition of reperfusion injury such as myocardial stunning and hemodynamic instability.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.82-82
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• 2002

Catalase(CAT)는 주요한 활성산소(ROS)의 일종인 과산화수소($H_2O$$_2$)가 Hydroxyl 유리기 생성에 관여하지 못하도록 $H_2O$$_2$를 $H_2O$ 및 $O_2$로 대사시켜 주는 효소계 항산화제의 한 종류이다. 따라서 본 실험에서는 CAT가 5% $O_2$및 20% $O_2$조건하에서 1-세포기 돼지체외수정란의 배발달에 미치는 영향을 조사하였다. 돼지난포란을 10% PFF, 0.1mg/ml cysteine, 10IU/m1 PMSG, 10IU/m1 hCG 및 10ng/m1 EGF가 첨가된 NCSU23 배양액에서 22시간 동안 배양을 실시하고, 성선자극호르몬이 배제된 배양액에서 추가로 22시간을 배양하여 체외성숙을 유도하였다. 체외성숙이 유기된 난자는 난구세포를 제거하고, 2.5mM caffeine과 0.1% BSA가 첨가된 mTBM 배양액에 정자를 1.25 $\times$ $10^{5}$cells/ml의 농도로 5-6시 동안 공동배양을 실시하여 체외수정을 유도하였다. 체외수정후 1-세포기의 수정란을 0.4mg/ml BSA가 첨가된 NCSU23 배양액에 CAT를 각각 0, 100, 500 및1,000uni1 첨가하여 30 embryos/ 50u1 소적으로하여 38.8$^{\circ}C$, 5% $CO_2$및 5%$CO_2$, 5% $O_2$90% $N_2$조건하의 배양기에서 각각 7일간 배양을 실시하였다. 조사된 결과는 SAS/STAT 6.03 Package를 이용하여 통계분석을 실시하였다. 체외배양 48시간에 난할률을 조사하였을 때, 대조구와 처리구간 차이가 인정되지 않았으나, 배양 7일째 배반포형성률에 있어서는 각각 22.7$\pm$2.7, 22.1$\pm$3.9, 18.7$\pm$4.9, 및 15.1$\pm$2.5%로서 처리구가 대조구보다 유의적으로 낮은 결과를 나타냈다. 그리고 산소농도에 따른 배반포형성률은 5% $O_2$ 조건(22.1$\pm$2.4%)이 20% $O_2$조건(17.2$\pm$2%)보다 유의적(P<0.05)으로 높은 것으로 나타났다. 총세포수에 있어서는 각각 44.4$\pm$4.0, 43.3$\pm$36, 25.4$\pm$2.4 및 13.4$\pm$1.5로서 처리구가 대조구보다 세포수가 적은 것으로 나타났으며, 산소농도에 따른 차이는 인정되지 않았다. 따라서, 체외생산된 돼지초기수정란을 배양할 때, CAT의 첨가는 효과가 없는 것으로 나타났다.다.