• 한국가축번식학회지
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• 제25권4호
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• pp.409-419
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• 2001

본 실험은 PF과 FPP가 소와 사람 동결 정자에 첨가되었을 때 응해 후 정자의 체외 생존성, 운동성 그리고 intact acrosome 향상에 기여할 수 있는지의 여부를 조사하고자 실시하였다. 사람의 정액은 TYB 동결 배양액을 사용하여 초 급속 동결하였다 PF과 FPP 첨가 효과는 각각의 시약이 동결-응해된 소나 사람 정자의 현미경적 조사에 의한 운동성에 미치는 영향과 Coomassie brilliant blue 염색방법에 의한 intact acrosome의 비율에 미치는 영향으로 조사하였다. Bovine 동결 응해 정자에 PF을 첨가하여 운동성을 조사하였던 바, 5 mM 처리군 (50.0%)이 대조군 (34.0%) 보다 유의하게 높은 운동성을 보여주었다 (F＜0.05). 동결 응해된 소 정자에 FPP를 농도에 따라 처리하여 intact acrosome을 조사하였던 결과, 50 nM 치리춘 (49%)이 대조군과 25 nH 처리군 (30.0, 38.0%) 보다 유의하게 많은 intact한 acrosome을 유지하였다 (P＜0.01). 사람 정자에서 동결에 앞서 PF을 농도에 따라 첨가하여 동결 응해 후 운동성을 조사한 결과, 5.0 mM 처리군 (51.0%)이 대조군과 2.5 mM (39.0, 40.0%) 처리군의 운동성보다 높았다 (P＜0.01). 사람 정액의 모든 동결 처리과정 (동결전, 동결, 응해후)에서 50 nM (75.5%) FPP 첨가가 intact acrosome percentage 유지하는데 유의한 효과가 있었다 (대조군: 45.0; 25 nM: 53.0; 100 nM: 68.0%) (P＜0.01). PF와 FPP 첨가하여 사람 정자의 동결융해 후 운동성과 intact acrosome에 미치는 영향을 동시에 비교해본 결과, 운동성에서는 PF 처리군이 약간 높지만 intact acrosome rate는 FPP 처리군의 결과 (65.0%)가 PF 처리군 (43.0%)보다 유의하게 높았다 (P＜0.05). 따라서 본 실험은 동결-융해된 소 정자에 PF이나 FPP 첨가는 정자의 운동성이나 intact acrosome 비율을 좀더 개선시킬 수 있고, 특히 사람 정자는 동결 전 과정에 FPP를 첨가하는 것이 정자의 체외 생존성, 운동성 그리고 intact acrosome을 유의하게 향상시킬 수 있다는 것을 시사한다.

• 지질공학
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• 제24권1호
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• pp.111-122
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• 2014

지반에 대한 정확한 이해를 위해 비교란 시료의 채취를 통한 실내실험이 필수적이며, 사질토 지반의 비교란 시료 채취는 인공동결공법에 의한 방법이 가장 효과적인 것으로 알려져 있다. 본 연구의 목적은 유사한 상대밀도를 가지는 동결-융해시료와 비동결시료의 비배수 삼축압축실험을 이용한 강도평가와 실험과정에서 측정한 압축파와 전단파 속도의 특성변화를 관찰하는 것이다. 인공동결공법에 의해 채취된 사질토 동결 시료를 모사하기 위해 주문진 표준사를 이용하여 수중강사법으로 60%와 80%의 상대밀도를 가지는 동결시료와 비동결시료를 조성하였다. 동결된 시료는 삼축압축실험용 페데스탈에 거치하여 자연융해하면서 1분 간격으로 시료의 온도를 측정하였다. 시료가 완전히 융해된 후 비동결시료와 동일한 방법으로 실험을 실시하였으며, 포화, 압밀, 전단과정에서 연속적으로 압축파와 전단파를 측정하여 속도를 산정하였다. 실험결과, 동결시료는 비동결시료에 비하여 축차응력과 전단강도는 감소하는 결과를 보였지만, 내부마찰각은 동결-융해의 여부와 상관없이 일정한 값을 나타내었다. 압축파 속도는 포화과정에서 B-value가 증가함에 따라 약 1800 m/s까지 증가하여 수렴하였으나, 압밀과정과 전단과정에서는 일정하게 유지되는 경향을 보였다. 전단파 속도는 포화과정에서 B-value가 증가함에 따라 감소하였고, 압밀과정과 전단과정에서는 시료가 받는 유효응력의 변화에 따라 거동하였다. 실험과정에서 압축파 속도는 상대밀도와 동결-융해여부에 상관없이 유사한 경향을 나타내었으나, 전단파 속도는 같은 상대밀도를 가지더라도 동결-융해시료가 비동결시료에 비해 작은 값을 나타내었다. 본 연구는 동결-융해시료와 비동결시료의 삼축압축실험 결과와 탄성파 특성을 비교함으로써 향후 인공동결공법으로 채취된 비교란 동결시료의 강도평가를 위한 예비실험으로 의의가 있다.

• Clinical and Experimental Reproductive Medicine
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• 제19권2호
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• pp.153-162
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• 1992

To solve the logistical problems of the sperm penetration assay (SPA) to provide just a sufficient number of hamster ova exactly when they are needed, a new method to cryopreserve the ova has been devised (1-step dehydration and 2-step thawing). After freezing & thawing of zona-intact (ZI) and zona-free (ZF) hamster ova according to this new method, the frozen-thawed ova were compared with fresh, control ova (FO) in terms of the degree of sperm penetration in SPA using semen samples from fertile donors, subfertile, and infertile male. Each sperm sample was capacitated for 42 hours inTEST-Yolk Buffer before insemination in SPA. In fertile doner, both the penetration rate and penetration index were lower in SPA using frozen ova (ZI; 92.4%, 6.2, ZF; 63.7%, 3.9) than those of SPA using fresh ova (99.3%, 8.4). There was a significant correlation between the penetration index of SPA using FO and ZI (p<0.001), and between those of SPA using FO and ZF and ova (p<0.001). In subfertile patient, both the penetration rate and penetration index were lowered in frozen ova (ZI; 62.3%, 1.3, ZF; 21.8%, 0.4) than those of fresh ova (74.8%, 1.8). There were significant correlation between the penetration rate and penetration index in ZI ova (p<0.05 and p<0.001, respectively). In infertile patient, both the penetration rate and penetration index were ZI; 3.1%, 0.0, ZF;0.0%, 0.0, respectively. There were significant correlation between the penetration rate and penetration index in ZI ova (p<0.05).

• 콘크리트학회논문집
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• 제19권6호
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• pp.701-706
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• 2007

본 연구는 장기간 숏크리트의 강도를 유지할 수 있는 영구지보재로서 알칼리 프리계 및 시멘트 광물계 급결제를 사용한 적정 배합비의 고성능 숏크리트의 내구성을 평가하였다. 내구성 실험은 염소이온 투과 시험, 동결융해 반복 시험, 촉진 중성화 시험 및 숏크리트에 염해의 영향 실험을 실시하였다. 실험 결과 실리카퓸을 사용한 모든 배합이 낮은 투과성을 나타내었다. 실험 결과 실리카퓸을 사용한 모든 배합이 모두 낮은 투과성을 나타내었다. 또한 동결융해 300회 반복 후에 상대동탄성계수가 85% 이상을 초과하는 우수한 동결융해 저항성을 보였다. 촉진 중성화 시험 결과 급결제에 따른 영향은 나타나지 않았으며 실리카퓸을 사용한 배합에서 중성화 깊이가 약간 깊게 나타났다. 염해 환경에서는 어떠한 영향도 없었다. 따라서 본 연구에서의 고성능 숏크리트 배합은 우수한 내구 성능을 보여주었다.

• 한국수정란이식학회지
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• 제23권4호
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• pp.239-243
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• 2008

This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at $4^{\circ}C$ for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into $37^{\circ}C$ water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.

• Journal of Pharmaceutical Investigation
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• 제33권4호
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• pp.293-298
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• 2003

This study was attempted to develop the physicochemical ad morphological stability test methods for the cream and lotion formulations among the cosmetic foundations and to provide the guidance for the stability methods with respect to basic emulsions and creams. With these developed stability test methods, we can evaluate the expired date or life time of the available basic cosmetics, especially basic lotions ad creams. Also, the stability test methods established in this study can be used as a guideline to test physical and morphological stability of cosmetics in the future. Thus, we selected two types of basic cosmetics such as lotions and creams made by four different cosmetic companies ad applied them to the stability test methods depending on the temperature changes such as temperature cycling and freezing-thawing cycling test. After the temperature changes, the conductivity, turbidity, particle size, creaming ratio and pH changes of the creams and lotions were evaluated and morphological changes such as crystal formation, odor, color and feeling of the creams and lotions were also tested. As the results of the stability tests, all the tested creams and lotions except for one lotion were stable. Therefore, it may be concluded that these short-term accelerated stability tests as physical stability test depending on the temperature change study were suitable for the stability testing methods for the basic cosmetics and may be useful for the establishment of the guideline for the stability test of cosmetics.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.831-837
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• 2005

Low-temperature adaptation and cryoprotection were studied in Leuconostoc mesenteroides SYl, a strain isolated from Kimchi. L. mesenteroides SY1 cells grown in exponential growth phase at $30^{\circ}C$ were exposed to $15^{\circ}C,\;10^{\circ}C$, and $5^{\circ}C$ for 2, 4, and 6 h, respectively, and then frozen at $- 70^{\circ}C$ for 24 h. Survival ratio was measured after the cells were thawed. The freezing-thawing cycles were repeated four times. Preadapted cells survived better than non-adapted control cells, and the highest survival ratio ($96\%$) was observed for cells preadapted for 2 h at $5^{\circ}C$, whereas control cells showed only $22\%$. The 2D gel showed that two proteins (spots A and B) were induced in cells preadapted at lower temperatures. Spots A and B have the same molecular weight (7 kDa), but the pI was 4.6 for spot A and 4.3 for spot B. The first 29 and 15 amino acid sequences from spots A and B were determined, and they were identical, except for one amino acid. A csp gene was cloned, and nucleotide sequencing confirmed that the gene encoded spot A cold shock protein.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.17-17
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• 2003

The milk of transgenic animals may provide an attractive vehicle for large-scale production of hEPO. Since glycosylation is cell type specific, recombinant human EPO (rhEPO) produced in different host cells contain different patterns of oligosaccharides, which could affect the biological functions. However, there have been no reports on the characteristics of rhEPO derived from milk of transgenic animals. To address this objective, several transgenic mice by using pWAPhEPO and/or pBC1hEPO expression vector were produced. However, 2 lines of pWAPhEPO founder female mouse died during late gestational day (day 18) before offspring could be obtained. They showed a severe splenomegaly, Unlike those of pWAPhEPO, mammary gland epithelial cells from biopsies of lactating pBC1hEPO transgenic mice had marked immunoreactivity to EPO and any activity was not detected in other tissues. The expression level of rhEPO is about 0.7% of mammary gland cellular total soluble proteins and an amount of 300~500 mg/L rhEPO is secreted into milk. Furthermore, the pBC1hEPO transgenic mice transmitted this character to their progeny in mendelian manner. In order to determine the extent of glycosylation variation, N-linked oligosaccharide structures present in the milk-derived rhEPO were characterized. Most of milk-derived rhEPO is fully glycosylated. the biological activity of milk-derived rhEPO was comparable to that of purified CHO-derived rhEPO, and milk-derived rhEPO showed relatively stable after freezing and thawing. Taken together, the results illustrate the potential of transgenic animals in the large-scale production of biopharmaceuticals.

• 한국수정란이식학회지
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• 제31권4호
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• pp.355-365
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• 2016

The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

• 한국농공학회지
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• 제24권2호
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• pp.67-75
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• 1982

A study on the effect of water reducing agent on the various characteristics of concrete has been conducted. The experimental results of the study are summarized as follows. 1. Slump test for the concrete added water reducing setretarding agent in proper quantity have been conducted. According to the test results, the decreasing rate of slump value become bigger than plain concrete with increase of the unit weight of cement and elapse of time 2. In case the proper quantity content of maximum compressive strength in Fig. 5 of water reducing set retarding agent is added, unit weight of water is decreased about 15% or so as compared with plain concrete. with the increase of water reducing set accelerating agent content unit weight of water is decreased much more, And other hand, amount of air entraining shows the increasing tendency with the increase of water reducing agent content. 3. The adding rate of water reducing agent which produce maximum strength shows that WR-CH and WR-SA which is water reducing set-starding agent is 0.2% and WR-CO is 0.5% and that WS-PO which is water reducing set accelerating agent is 0.5 4. compressive strength jof the concrete made of sulfate resistant cement shows less than the strength of normal portland cement at initial strength but the strength of both cement shows almost same at curing age of 28 days. 5. when proper quantity of water reducing set retarding agent is used, boned strength is increased about 15% at curing age of 28days. 6. According to the result of durability test, dynamic young's mudulus of elasticity at plain concrete is decreased about 50% as compared with initial step at 300 cycle of freezing and thawing after curing age of days. on the contarary the concrete used water reducing agent is decreased less than 7%.