• Integrative Medicine Research
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• 제6권1호
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• pp.12-18
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• 2017

Cryopreservation is a process that preserves organelles, cells, tissues, or any other biological constructs by cooling the samples to very low temperatures. The responses of living cells to ice formation are of theoretical interest and practical relevance. Stem cells and other viable tissues, which have great potential for use in basic research as well as for many medical applications, cannot be stored with simple cooling or freezing for a long time because ice crystal formation, osmotic shock, and membrane damage during freezing and thawing will cause cell death. The successful cryopreservation of cells and tissues has been gradually increasing in recent years, with the use of cryoprotective agents and temperature control equipment. Continuous understanding of the physical and chemical properties that occur in the freezing and thawing cycle will be necessary for the successful cryopreservation of cells or tissues and their clinical applications. In this review, we briefly address representative cryopreservation processes, such as slow freezing and vitrification, and the available cryoprotective agents. In addition, some adverse effects of cryopreservation are mentioned.

• 한국농공학회지
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• 제33권2호
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• pp.112-119
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• 1991

Some experiments were carried out to investigate the effects of freezing and thawing on the strength and strain characteristics of alluvial silty clay under the different temperatures, loading and moisture conditions. The results were as follows; 1. The soil used was proved to be consisted of silty clay with honey-combed structure, and showed higher dilatancy, frost activity and lower stability in natural state. 2. Soil treated with freezing and thawing cycles showed lower compressive strength compared with the non treated, The strength decreased with incement of freezing and thawing cycles. It's shapes of stress-strain curves were flat and did not formulate a peak while the peak strength of higher moisture content soil decreased with the increment of moisture content. It's decrement ratio was most distinctly shown at the first one cycle of freezing and thawing. 3. The cohesion decreased due to freezing and thawing cycles but internal frcition angle was not changed. 4. The liquid limit decreased with increment of freezing and thawing cycles, and became almost constant after three cycles of freezing and thawing. 5. The strength under simple loading at failure mode was appeared to be higher compared with the cyclic loading after freezing and thawing but initial moisture content effect was not observed. 6. Ice lense was not observed within 50% of ice content ratio but observed over 100%. The higher the ice content ratio, the higher the peak strength. As a matter of fact, it seems that an optimum ice content ratio exists for plastic mode and the least compressive strength.

• 한국가금학회지
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• 제41권4호
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• pp.261-270
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• 2014

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결 보존 기술의 향상에 의해 동결 및 융해 후의 많은 생존 세포를 확보하는 것과, 생식계열 키메라의 제작 효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5~6일령의 닭 원시 생식선으로부터 채취하고, MACS 방법에 의해서 순수 닭 PGCs를 분리했다. 15% 각각의 EG와 DMSO를 동결 보호제로 사용한 처리군이 각 군의 농도에 상관없이 유의적(p<0.05)으로 PG 처리군보다 동결 및 융해 후의 세포의 생존율이 높음을 확인하였다. 특히, 10% EG+FBS 조합의 처리군에서 상업용 닭인 한협육종협회3호종(B : 88.7%)이 오계종(A : 85.1%), 아사 브라운종(C : 84.6%) 그리고 화이트 레그혼종(D : 85.9%)의 세 품종보다 동결 및 융해 후의 닭 PGCs의 생존율이 유의적(p<0.05)으로 높음을 확인하였다. 간이 동결에 있어서 가장 높은 생존율을 보인 10% EG이 10% DMSO와 함께 최적의 동결 보호제로서 사용 가능성을 확인하였다.

• Journal of Communications and Networks
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• 제12권1호
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• pp.43-51
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• 2010

IEEE 802.11 defines distributed coordination function (DCF), which is characterized by CSMA/CA and binary exponential backoff (BEB) algorithm. Most modifications on DCF so far have focused on updating of the contention window (CW) size depending on the outcome of own frame transmission without considering freezing periods experienced in the backoff interval. We propose two simple but novel schemes which effectively utilize the number of freezing periods sensed during the current backoff interval. The proposed schemes can be applied to DCF and its family, such as double increment double decrement (DIDD). Saturation throughput of the proposed schemes is analyzed by means of Bianchi's Markovian model. Computer simulation validates the accuracy of the analysis. Numerical results based on IEEE 802.11b show that up to about 20% improvement of saturation throughput can be achieved by combining the proposed scheme with conventional schemes when applied to the basic access procedure.

• 한국수정란이식학회지
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• 제29권1호
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• Journal of Microbiology
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• 제45권6호
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• pp.593-596
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• 2007

In this study, we describe the development of a simple and efficient method for cell lysis via the insertion of a bacteriophage lambda lysis gene cluster into the pET22b expression vector in the following order; the T7 promoter, a gene for a target protein intended for production, Sam7 and R. This insertion of R and Sam7 into pET22b exerted no detrimental effects on cellular growth or the production of a target protein. The induction of the T7 promoter did not in itself result in the autolysis of cells in culture but the harvested cells were readily broken by freezing and thawing. We compared the efficiency of the cell lysis technique by freezing and thawing to that observed with sonication, and determined that both methods completely disintegrated the cells and released proteins into the solution. With our modification of pET22b, the lysis of cells became quite simple, efficient, and reliable. This strategy may prove useful for a broad variety of applications, particularly in experiments requiring extensive cell breakage, including library screening and culture condition exploration, in addition to protein purification.

• 동력기계공학회지
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• 제7권2호
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• pp.17-22
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• 2003

As side-by-side refrigerators came into existence, there has been a growing concern about the free%ins-up of the vital equipment in a walk-in freezer. Due to a bad performance, customers are experiencing too much frustration. In order to minimize the freezing phenomena, the numerical simulation has been performed on the characteristics of cold air flow in a side-by-side refrigerator. The flow field has been simulated with a standard $k-\varepsilon$ turbulent model and a SIMPLE algorithm based on the finite volume method. Through the results of the analysis of the pattern of cold air flow, finally the shape of outlet for cold air flow was modified. The present model was compared with the modified model. The latter was better than the former in minimizing the freezing phenomena.

• 한국터널지하공간학회 논문집
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• 제16권5호
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• pp.431-443
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• 2014

본 연구는 고수압 해저터널의 차수를 위한 동결공법 적용성 평가를 위해 국내 적용사례가 없는 동결공법의 해외 시공사례를 조사하였으며, 지반 동결공법에서 보편적으로 사용하는 냉매인 브라인(brine)과 액체질소($LN_2$)에 대한 분석을 수행하였다. 고수압 조건의 해수는 순수한 물에 비하여 어는 점이 더 낮기 때문에 지상에서보다 동결시간이 더 길 수 있고 해저터널과 같이 폐쇄된 공간에서의 적용 시 냉매는 안정성을 확보할 수 있어야 하고 공기 단축이 가능해야 한다. 브라인은 재사용이 가능하고 독성이 적어 인체에 미치는 영향이 적으나, 동결에 상대적으로 장시간 필요하고 동결장비가 복잡한 특징을 가지고 있으며, 액체질소는 냉매의 재사용이 불가하며 기화 시 공기 중 질소농도 증가로 질식의 위험성이 있으나, 상대적으로 동결시간이 짧으며, 동결장비가 간단한 특징을 가지고 있는 것으로 파악되었다. 또한 향후 추가적인 연구가 필요한 해저터널 연결구 및 막장면 주변 지반에 대한 동결공법의 주요 설계요소를 도출하였다.

• Clinical and Experimental Reproductive Medicine
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• 제17권1호
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• pp.81-85
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• 1990

We cryopreserved mouse morulae by a simple ultra-rapid method of freezing embryos directly in $LN_2$ after holding 2min in a $LN_2$ vapor, and thawed them in $37^{\circ}C$ water bath. The time requirements for permeation and dehydration by 2.0 M glycerol and 0.2 M sucrose before freezing were studied. When the embryos were equilibrated for 10 min, the optimun post-thaw survival was obtained. Embryos those developed normally to blastocyst after in vitro culture for over 24hrs were regarded as survival ones. Two experiments to assess post-thaw survival following predehydration in various mixtures of glycerol and sucrose were also accomplished. When sucrose was held constant (0.2 M) and glycerol concentration varied (1.5-3.5 M), post-thaw survival was best (78.0%) in 3.0 M glycerol. When glycerol was held constant (3.0M) and sucrose concentration varied (0.0-1.0M), optimun post-thaw survival (78.0%) was found in 0.2 M sucrose.

• 한국도로학회논문집
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• 제15권3호
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• pp.53-63
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• 2013

PURPOSES : This study is to evaluate moisture susceptibility of asphalt mixtures by using non-destructive impact wave and to determine durability so as to decrease the gap between before and after freezing in the future. METHODS : Using non-destructive impact wave, this study is to determine the dynamic modulus of asphalt specimen. Furthermore, the results obtained from two experiment accelerometers are used for the dynamic modulus determination. The dynamic moduli of specimens are compared with those of the freezing-thawing specimens. RESULTS : Test results showed that the dynamic modulus before freezing and thawing environment loads at each temperature dropped about 3.7% after the environmental loads. Furthermore, correlation analysis indicates that transition of dynamic modulus at each point is about 89.59%. CONCLUSIONS: Evaluation of asphalt mixtures using non-destructive impact wave has excellent repeatability and simple equipment for the test. Consequently, the method in the study will be useful for evaluating the characteristics of a various asphalt mixtures.