• 한국동물위생학회지
• /
• 제18권3호
• /
• pp.1-12
• /
• 1995

To investigate effect of seeding on post-thaw motility and viability of canine spermatozoa, the semen from male dogs which had been proved to be fertile in the past were frozen and seeded during freezing process. Post-thaw motility and viability of canine sperm which were frozen and seeded were investigated according to different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$, or $-l5^{\circ}C$ and also according to different concentration of glycerol of 2%, 5% and 10%. In addition, post-thaw motility of canine sperm frozen by direct freezing in a deep freezer or programmed freezing in a programmed cell freezer was investigated. Post-thaw motility of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}C$ showed considerably higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, respectively, in 2% and 5% glycerol groups on both 2 and 7day after freezing(p<0.05). In 10% concentration of glycerol, the sperm seeded at each seeding temperature showed considerably higher post-thaw motility than that of non-seeding group on day 7 after freezing(p<0.01). Post-thaw viability of canine sperm was compared according to different seeding temperatures : The sperm seeded at $-5^{\circ}$ showed significantly higher post-thaw motility than that of non-seeding, and that seeded at $-10^{\circ}C$, or $-l5^{\circ}C$, in 5% and 10% glycerol groups on day 7 after freezing(p< 0.05). In comparison of post-thaw motility of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed considerably higher post-thaw motility than 2% glycerol group without difference between those two groups in all seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$) on day 2 and 7 after freezing(p<0.01). In comparison of post-thaw viability of canine sperm seeded according to different concentration of glycerol, 5% glycerol group and 10% glycerol group showed the same considerably higher post-thaw viability than 2% glycerol group on each thawing day(p<0.01). The canine sperm frozen and seeded by programmed freezing method showed considerably higher post-thaw motility than that frozen by direct freezing method in all different seeding temperatures($-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}$). These results indicated that the higher post-thaw motility and viability was obtained in the spermatozoa seeded than that of non-seeding, that among different seeding temperatures of $-5^{\circ}C$, $-10^{\circ}C$ and $-l5^{\circ}C$, the sperm seeded at $-5^{\circ}C$ showed higher post-thaw motility and viability than the other temperatures, also among different concentrations fof glycerol of 2%, 5% and 10%, the sperm frozen and seeded in 5% and 10% concentration of glycerol showed higher post-thaw motility and viability than that in 2% of glycerol, and that the sperm frozen and seeded by programmed freezing method showed higher motility than that by direct freezing method.

• 한국도로학회논문집
• /
• 제12권3호
• /
• pp.71-77
• /
• 2010

노상토의 동상 가능성이 없는 경우 동상방지층을 생략한다. 이러한 조건에서 노상토의 동결 융해가 발생하면, 동상이 없는 상태에서 탄성계수 변화를 수반하고, 포장설계에서 이를 합당하게 고려해야 한다. 동결 융해 과정의 연속적인 탄성계수 변화를 효과적으로 규명하고자 비파괴시험인 충격공진시험(IR, Impact Resonance)을 도입하였다. 동결 융해 과정에서 수분의 공급이 차단된 폐쇄형 동결조건에서 노상토의 탄성계수는 동결 융해 반복횟수에 무관하게 일정한 값으로 나타났다. 또한 시험에 적용한 모든 함수비 및 다짐도 조건에서 동결전의 탄성계수와 동결 융해 후의 탄성계수는 변화가 거의 없이 일정한 값으로 나타났다. 즉 폐쇄형 동결 융해 조건에서는 노상토의 융해 강성도 감소(thaw-weakening)가 발생하지 않았다. 동결시 탄성계수는 다짐도 및 함수비에 따라서 변화하였으나, 포장설계 관점에서는 무시 가능한 수준으로 나타났다.

• 한국수정란이식학회지
• /
• 제13권1호
• /
• pp.1-9
• /
• 1998

In order to improve the cryopreservation by vitrification or slow freezing of nuclear transplant rabbit embryos, the effects of factors affecting embryo cryopreservation such as cryoprotectants, equilibration, cooling rate and post-thaw dilution on post-thaw survial and development were determined using intact embryos of morular stage. And the post-thaw development of nuclear transplanted embryos cryopreserved under the optimal conditions examined was compared between vitrification and slow freezing. The cryoprotectant solution used was ethyleneglycol-ficoll-sucrose (EFS) or ethyleneglycol-poly-vinylpyrrolidone-galactose- I (EPG- I ) for vitrification, and EPG- II for slow freezing. To examine the viability of frozen-thawed embryos, the nuclear transplanted embryos were co-cultured in TCM-199 plus 10% FBS with bovine oviduct epithelial cells(BOEC) for 24 hrs and the intact morulae were co-cultured with BOEC for 5 days and 3 days to hatching blastocyst stage in 39 ˚C 5% $CO_2$ incubator. The results obtained were as follows: Following vitrification with EFS, the post-thaw development of rabbit morulae to hatching blastocyst was significantly(P<0.05) higher in compacted stage(82.4%) than in early morular stage(60.0%). The post-thaw development of compacted morulae to hatching blastocyst was similarly high in vitrification with EFS(82.4%), EPG- I (85.0%) and in slow freezing with EPG- II (83.3%). Following vitrification with EPG- I, the post-thaw development of intact rabbit morulae to hatching blastocyst was similar as 78.0% and 85.0% in 1-step and 2-step post-thaw dilution, respectively. The post-thaw development of nuclear transplanted rabbit embryos of compacted morulae stage to hatching blastocyst was similarly 43.6% and 40.0% in vitrification with EPG- Iand slow freezing with EPG- II, respectively. These results indicated that the rabbit nuclear transplant and intact embryos of morulae stage could be well cryopreserved with either vitrification or slow freezing procedure.

• Geomechanics and Engineering
• /
• 제15권4호
• /
• pp.997-1004
• /
• 2018

The brine leakage is a tough problem in artificial freezing engineering. This paper takes the common soft clay in Wujiang District as the study object, and calcium chloride solutions with six salinity levels were considered. The 'classic' cooling curve method was employed to measure the freezing point of specimens after freeze-thaw. Results indicate that four characteristic stages can be observed including supercooling, abrupt transition, equilibrium and continual freezing, strongly dependent on the variation of unfrozen water content. Two characteristic points were found from the cooling curves, i.e., freezing point and initial crystallization temperature. A critical value for the former exists at which the increment lowers. The higher the saline content approximately linearly, lower the freezing point. In the initial five cycles, the freezing point increases and then stabilizes. Besides, the degree of supercooling was calculated and its correlations with water, salt and freeze-thaw cycles were noted. Finally, an empirical equation was proposed for the relationship of freezing point and three main factors, i.e., water content, saline content and freeze-thaw cycles. Comparison of calculated and measured data proves that it is reliable and may provide guidance for the design and numerical analysis in frozen soil engineering.

• 한국임상수의학회지
• /
• 제17권2호
• /
• pp.420-430
• /
• 2000

These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

• 한국농공학회지
• /
• 제31권3호
• /
• pp.70-80
• /
• 1989

In this research program special triaxial compression tests and dehydration-swelling tests under the condition of freeze-thaw process were conducted to show the effects of freezethaw process on the physical properties of weathered granite soil, and their results as follows; 1.Consolidation settlement of weathered granite soil mass was increased due to freeze-thaw process, and the initial tangent coefficient of dense state was higher than that of loose state. 2.Compression behaviour of soil was increased according to the decrease of freezing temperature, and when the freezing temperature was reached under - 10$^{\circ}$C, the compression rate was not influenced by change of freezing temperature. 3.The experiments showed that the void ratio and permeability of soil were converged into their values of shrinkage limit, and the permeability was higher due to the freeze-thaw process and as the lower the freezing temperature. 4.The decrease of liquid limit was indicated as the lower the freezing temperature, and as more the freeze4haw cycles, the moisture content was shown the lower side. 5.It was shown that the shrinkage was decreased by freeze-thaw process and not influenced by way of freezing temperature, but dehydration rate was higher.

• 한국발생생물학회지:발생과생식
• /
• 제2권1호
• /
• pp.63-68
• /
• 1998

본 연구에서는 보다 효율적인 동결 보존법을 수립하기 위하여 현재 사용되는 동결 보존액과 동결방법을 정자의 운동성 측면에서 비교해 보았다. 즉, 세 종류의 조성이 다른 동결보존액인 TYB, dithiothreitol을 첨가한 TYB+DTT, KS II 등이 동결보존 전후에 있어 운동성에 미치는 영향을 조사하였으며, 또한 vapor freezing 방법과 computerized freezer를 사용한 동결방법이 정자 운동성에 미치는 영향을 알아보았다. 정자의 분석은 현미경적 방법과 두 종류의 컴퓨터 정자 자동측정기인 SAIS와 Hamilton Thorn을 사용하여 동결 전 후의 정자 운동성과 VCL, VSL, VAP, ALH, LIN 등의 sub-motility 패턴을 측정하였다. 정액성상이 정상인 군에서 동결보존액을 비교한 실험결과는 TYB군과 TYB+DTT군, 그리고 KS II군의 융해 후 운동성이 각각 28.3%, 23.0%, 34.8%로 KS II군이 우수하였고, 동결방법을 비교한 실험에서는 vapor freezing 군과 computerized freezing 군의 융해 후 정자 운동성이 각각 27.8%, 33.2%로 유의차는 없었다. 또한 무력정자증을 보인 정액군에서는 TYB군과 TYB+DTT군, 그리고 KS II군에서 융해후 정자 운동성이 각각 13.6%, 10.0%, 18.5%로 여기 KS II군이 우수하였으며, vapor freezing군과 computerized freezing군의 융해 후 정자 운동성은 12.8%, 12.9%로 유의차가 없었다. 이상의 결과로 보아 운동성이 정상인 정액군과 무력정자증을 보이는 정액군에서 KS II를 사용해 동결하는 것이 TYB나 TYB+DTT를 사용하는 것보다 운동성 있는 정자를 회수하는데 더 효율적이며, 동결방법 측면에서는 vapor freezing 방법과 computerized freezing방법간에 큰 차이가 없음을 볼 수 있었다.

• Asian-Australasian Journal of Animal Sciences
• /
• 제22권3호
• /
• pp.343-349
• /
• 2009

This study was designed to simplify a cryopreservation program for embryonic stem cells (ESCs) by selection of cooling method and cryoprotectant. Commercially available mouse E14 embryonic stem cells (ESCs) were cryopreserved with various protocols, and morphology and viability of the frozen-thawed ESCs and their reactive oxygen species (ROS) production were subsequently monitored. Post-thaw colony-formation of ESCs was detected only after a slow freezing using dimethyl sulfoxide (DMSO) by stepwise placement of a freezing container into a $-80^{\circ}C$ deep freezer and subsequently into -$196^{\circ}C$ liquid nitrogen, while no proliferation was detected after vitrification. When the simplified protocol was employed, the replacement of DMSO with a mixture of DMSO and ethylene glycol (EG) further improved the post-thaw survival. ROS generation in ESCs frozen-thawed with the optimized protocol was not increased compared with non-frozen ESCs. The use of fresh mouse embryonic fibroblasts as feeder cells for post-thaw subculture did not further increase post-thaw cell viability. In conclusion, a simplified slow-freezing program without employing programmable freezer but using DMSO and EG was developed which maintains cell viability and colony-forming activity of ESCs during post-thaw subculture.

• 한국도로학회논문집
• /
• 제18권6호
• /
• pp.123-130
• /
• 2016

OBJECTIVES : The objective of this research is to determine the moisture resistance of the freeze-thaw process occurring in low-noise porous pavement using either hydrated-lime or anti-freezing agent. Various additives were applied to low-noise porous asphalt, which is actively paved in South Korea, to overcome its disadvantages. Moreover, the optimum contents of hydrated-lime and anti-freezing agent and behavior properties of low-noise porous asphalt layer are determined using dynamic moduli via the freeze-thaw test. METHODS : The low-noise porous asphalt mixtures were made using gyratory compacters to investigate its properties with either hydrated-lime or anti-freezing agent. To determine the dynamic moduli of each mixture, impact resonance test was conducted. The applied standard for the freeze-thaw test of asphalt mixture is ASTM D 6857. The freeze-thaw and impact resonance tests were performed twice at each stage. The behavior properties were defined using finite element method, which was performed using the dynamic modulus data obtained from the freeze-thaw test and resonance frequencies obtained from non-destructive impact test. RESULTS : The results show that the coherence and strength of the low-noise porous asphalt mixture decreased continuously with the increase in the temperature of the mixture. The dynamic modulus of the normal low-noise porous asphalt mixture dramatically decreased after one cycle of freezing and thawing stages, which is more than that of other mixtures containing additives. The damage rate was higher when the freeze-thaw test was repeated. CONCLUSIONS : From the root mean squared error (RMSE) and mean percentage error (MPE) analyses, the addition rates of 1.5% hydrated-lime and 0.5% anti-freezing agent resulted in the strongest mixture having the highest moisture resistance compared to other specimens with each additive in 1 cycle freeze-thaw test. Moreover, the freeze-thaw resistance significantly improved when a hydrated-lime content of 0.5% was applied for the two cycles of the freeze-thaw test. Hence, the optimum contents of both hydrated-lime and anti-freezing agent are 0.5%.

• Asian-Australasian Journal of Animal Sciences
• /
• 제17권9호
• /
• pp.1291-1295
• /
• 2004

Histology and transmission electron microscopy studies were carried out on buffalo muscles that were subjected to repeated freeze-thaw cycles at -10 and $-18^{\circ}C$. In the first freeze thaw cycle ($-10^{\circ}C$) structures of muscle showed slight change and closely resembled to those of normal muscle. There were frequent gaps in the half way across the fibres and some cracks in individual fibre were also noticed in second freeze thaw cycle. In the muscle frozen at $-18^{\circ}C$, more pronounced shrinkage with extensive damage of fibres with tearing was observed. The interfibrillar gaps were wider, shrinkage and tearing of the fibres were more distinct after second freeze-thaw cycle. After the second cycle, the interior portion showed large scale degradation of the ultrastructure. Our studies of buffalo muscle showed that under the proper condition, little structural damage takes place in the meat histology and ultrastructure under repeated freeze-thaw conditions. This study adds continued weight to the evidence that limited freeze-thaw cycles will not deteriorate the quality of meat.