• Title/Summary/Keyword: Slow-freezing

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Study on Cryopreservation of Epididymal and Ejaculated Semen in Korean Native Canine and Subsequent Pregnancy Rate after Artificial Insemination

  • Kim S. K.;Lee B. K.;Kim M. K.
    • Reproductive and Developmental Biology
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    • v.28 no.3
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    • pp.155-159
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    • 2004
  • This study was carried out to investigate the general characteristics and viability of sperm after freezing and thawing and the pregnancy rates after artificial insemination with thawed semen. The rates of viable sperm after slow and rapid freezing were 87.4±3.85% and 70.8±4.45%, respectively which were significantly lower than that of fresh semen control (91.7±3.45%). The mean concentration of epididymal sperm after dilution in 1.0 ml saline and. 3.0 ml extender in a various concentrations of cryoprotectants was 124.5±48.3 x 10/sup 6/ (range of 45 x 10/sup 6/ to 280 X 10/sup 6/ /ml). There was a significant difference not in the percentage of acrosome-reacted sperm, but in the percentage of capacitated sperm, between fresh and frozen-thawed epididymal semen. When frozen-thawed after diluting with tris-buffer extender containing glycerol, DMSO and ethylene glycol with concentration of 2 to 6%, the rates of epididymal sperm exposed to different cryoprotectants ranged from 14.4±4.7% to 20.7±5.8%, 17.8±5.2% to 36.5±4.9%, and 14.4±4.6% to 18.5±5.3%, respectively which were lower compare to fresh semen control. The pregnancy rate after artificial insemination with frozen semen was 70.6%, whereas that with fresh semen was 90.0% in dogs with naturally induced estrus.

Cryopreservation (Vitrification) of Mouse Embryos (마우스의 배의 동결보존)

  • 강민수
    • Journal of Embryo Transfer
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    • v.6 no.2
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    • pp.30-36
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    • 1991
  • The method of vitnilcation has various merits. It needs neither seeding nor slow freezing. It can freeze embryo by putting it directly into liquid nitrogen at the indoor temperature to $0^{\circ}C$. The operation process is quite easy. Moreover, higher promise of survival can be expected as there is no physical damage by any lumps of ice with the exception of cells. In Kasal's experiment (1990) using EFS liquid and Kang's experiment (1991) using GFS liquid the ratio of the damaged embryo was only 2-3%. But, the method of vitrification is now on the process of improvement, and the final or united method is not yet established. At the present time, most of the major institutes all over the world are using the traditional freezing method in the preservation of mouse embryo, but it is very likely that the vitrification will prevaIl in the near future considering the various merits of it. Calves can be begotten from the embryo by means of vitriilcated preservation in the cases of cow, rat, and rabbit as well as of mouse. In addition, recent experiments have shown that vitrificated preservation was successful in the case of drosophila embryo which was much bigger than mammalian embryo, which fact tells that this method is expected to be preferably used even in the preservation of living organs in the near future.

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A Study on the Strength Properties and the Temperature Hysteresis of Winter Concrete according to the difference of Curing Method in Mock-up Test (실물대시험에서의 양생방법 차이에 따른 한중콘크리트의 온도이력 및 강도특성에 관한 연구)

  • Won, Cheol;Han, Cheon-Goo
    • Journal of the Korea Institute of Building Construction
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    • v.3 no.4
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    • pp.87-94
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    • 2003
  • This study is to investigate the temperature hysteresis and development of compressive strength due to the curing conditions and to evaluate the optimum curing condition of test specimens showing the same development of strength to that of real structures in cold weather. The results of temperature curve with curing conditions in mock-up tests showed the trend of decrease plain concrete with insulation form, plain concrete with heating, concrete with accelerator for freeze protection, and control concrete in turn. The strength development of plain concrete of inside and outside of shelter showed the very slow strength gains due to early freezing, but that of concrete with accelerator for freeze protection showed the gradual increase of strength with time. From this, it is clear that accelerator for freeze protection has the effects of refusing the freezing temperature and accelerating the hardening under low temperature. Strength test results of small specimens embedded in members and located in insulation boxes at the site are similar to that of cores drilled from the members at the same ages, thus it is clear that these curing methods are effective for evaluating in-place concrete strength

Development of Temperature Control Algorithm for Supercooling Storage of Pork Loin and Its Feasibility for Improving Freshness and Extending Shelf Life

  • Lee, SangYoon;Park, Dong Hyeon;Kim, Eun Jeong;Kim, Honggyun;Lee, YunJung;Choi, Mi-Jung
    • Food Science of Animal Resources
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    • v.42 no.3
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    • pp.467-485
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    • 2022
  • Supercooling storage refers to lowering the product temperature below its freezing point without phase transition and has the potential to extend shelf life. Nevertheless, supercooled objects are in a thermodynamically unstable state, and nucleation can occur spontaneously. To achieve supercooling storage, slow cooling and insulation are essential. Hence, a stepwise algorithm for the supercooling storage of pork loins was designed and validated in this study. Pork loins were stored at 3℃, -18℃, and -3℃ (freezing), and supercooled for 16 days. All samples remained in a supercooled state and were unfrozen at the end of storage. Supercooled pork loins were superior in terms of drip loss, cooking loss, and water-holding capacity compared to frozen samples. Additionally, supercooling treatment prevented discoloration, increase of volatile basic nitrogen, and microbial growth. Thus, supercooling of pork loin was achieved using a stepwise program and was effective to maintain meat quality.

Cryopreservation of Human Immature Follicular Oocyte (인간의 미성숙난자의 동결보존에 관한 연구)

  • Kim, Eun-Kyung;Son, Weon-Young;Chi, Hee-June;Ko, Jung-Jae;Yoon, Tae-Ki;Cha, Kwang-Yul
    • Clinical and Experimental Reproductive Medicine
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    • v.19 no.2
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    • pp.163-168
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    • 1992
  • This study was carried out to set up the ovum bank for ovum donation and to determine the best freezing method for human immature oocytes. Human immature follicular oocytes were cryopreserved by slow freezing and rapid thawing method. Immature follicular oocytes were treated by propanediol(PROH) solution by 2 and 4 step method in protocols A & B, respectively. In protocol C, immature oocytes were exposed to sucrose prior to treatment of PROH by 4 step method. We compared survival rate, maturation rate, and fertilization rate of immature oocytes among three protocols. Results were as follows. 1. Oocytes treated by the protocol C showed the highest survival rate( 70.3 %) and maturation rate(34.6%) after thawing. 2. Survival rate of oocytes treated by the protocol C was significantly higher than that of the protocol B after thawing(p<0.05). In conclusion, treatment of oocytes with sucrose prior to expose PROH was the best freezing method. Sucrose may have reduced the toxic effect of cryoprotectant to oocytes. We failed to induce fertilization of oocytes, which were treated by any protocols, by conventional insemination method, but obtained 28.8% fertilization rate by using partial zona dissection(PZD) method. This result suggests that micromanipulation(PZD) of the thawed oocytes before insemination will improve the fertilization rate.

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Cryopreservation of Porcine Embryos using Glycerol, Egg Yolk and Trehalose (Glycerol, 난황 및 Trehalose를 이용한 돼지유정란의 동결)

  • 장원경;박수봉;이명식;김태현;박용윤;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.19 no.1
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    • pp.43-48
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    • 1995
  • The purpose of this study was to examine the suvival rates of cryopreserved porcine embryos collected on Day 6 (Day 0=onset of estrus) with various cryprotectants. Eighty two embryos at different stages, ranged from expanded blastocyst to hatched blastocyst, were allocated to 6 experimental groups in different combinations of cryoprotectants glycerol, egg yolk and/or trehalose. Porcine embryos were cryopreserved using conventinal slow freezing precedures. The embryos were equilibrated with one of the freezing solutions, cooled from 25 to -7$^{\circ}C$ at 1$^{\circ}C$/min, seeded at -7$^{\circ}C$ frozen to -36$^{\circ}C$ at 0.5$^{\circ}C$/min, and then plunged into liquid nitrogen. The frozen embryos were thawed by immersion in 37$^{\circ}C$ water and the cryoprotectants were removed by dilution with 0.5M sucrose solution. Embryonic survival was estimated from the normal development of embryos for 12, 24 and 48hrs culture. Then the embryos were stained and their cell nuclei was counted. The survival rates of morphological embryos were significantly higher in group I (10% glycerol) or group IV (10% glycerol+10% egg yolk+0.5M trehalose) than those in other groups, although the nuclei number was quite higher in embryos treated with 10% glycerol and 0.25M trehalose at expended blastocyst. However, hatched blastocysts showed higher viability and nuclei number treated with either egg york or trehalose, but the survival rates after 48hrs of culture were quite low. These results indicate that egg yolk and trehalos as a supplement to freezing solutions can be useful to the cryopreservation ofn porcine embryos.

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A Hetero-Mirroring Scheme to Improve I/O Performance of High-Speed Hybrid Storage (고속 하이브리드 저장장치의 입출력 성능개선을 위한 헤테로-미러링 기법)

  • Byun, Si-Woo
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.11 no.12
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    • pp.4997-5006
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    • 2010
  • A flash-memory-based SSDs(Solid State Disks) are one of the best media to support portable and desktop computers' storage devices. Their features include non-volatility, low power consumption, and fast access time for read operations, which are sufficient to present flash memories as major database storage components for desktop and server computers. However, we need to improve traditional storage management schemes based on HDD(Hard Disk Drive) and RAID(Redundant array of independent disks) due to the relatively slow or freezing characteristics of write operations of SSDs, as compared to fast read operations. In order to achieve this goal, we propose a new storage management scheme called Hetero-Mirroring based on traditional HDD mirroring scheme. Hetero-Mirroring-based scheme improves RAID-1 operation performance by balancing write-workloads and delaying write operations to avoid SSD freezing. Our test results show that our scheme significantly reduces the write operation overheads and freezing overheads, and improves the performance of traditional SSD-RAID-1 scheme by 18 percent, and the response time of the scheme by 38 percent.

Long-term Durability Characteristics of Fly ash Concrete Containing Lightly Burnt MgO Powder (저온 소성한 MgO 분말을 함유한 플라이애시 콘크리트의 장기재령에서의 내구특성)

  • Jang, Bong-Seok;Choi, Seul-Woo;Lee, Kwang-Myong
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.33 no.3
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    • pp.909-916
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    • 2013
  • Concrete containing lightly burnt MgO has long term expansibility. It also could compensate for the thermal shrinkage of mass concrete, because the hydration of MgO proceeds at a slow pace to long-term age. Thus, lightly burnt MgO has been applied to the construction of mass concrete such as dams. Recently, the expansion characteristics of MgO concrete with fly ash that could be applied to mass concrete for the reduction of hydration heat have been studied and however, limited studies on its durability. This study investigates the long-term durability characteristics of fly ash concrete with lightly burnt MgO. The durability tests on carbonation, freezing-thawing, diffusion of chloride, and resistance to sulfate attack were carried out for MgO concrete with curing for 360 days in submerged condition with different temperature of 20 and $50^{\circ}C$. The results reveal that MgO concrete shows a greater resistance of carbonation, diffusion of chloride, and resistance to sulfate attack. On the other hand the resistance of freezing-thawing was little influenced by MgO powder.

The Experimental Study on Cryopreservation of Mouse Embryo (생쥐배아의 동결보존에 관한 실험적 연구)

  • Lee, Yu-Il;Kwon, Young-Sook;Park, Hyun-Jeong
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.1
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    • pp.55-63
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    • 2001
  • Objectives: This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Materials and Methods: Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Two-step 1,2-propanediol (PROH), dimethylsulfoxide (DMSO) and 4-step PROH DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. Results: As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in PROH and DMSO was significantly higher than 4-8 cell (64.5% versus 62.1 %,79.7% versus 73.2%) (p<0.01, p<0.01), but the development rates of 4-8 cell embryos in PROH and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4-8 cell embryos in PROH were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in PROH (74.4% versus 64.5%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The developmen1 rate from morular to hatching blastocyst, however, was significantly higher in PROH than in DMSO during 48 hr (p<0.01). The survival rate of 4-8 cell embryo was 62.1% in PROH and 73.2% in DMSO. The development rates of embryo in PROH were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. Conclusions: The survival rate of embryo in 2 cell stage was higher than in 4-8 cell stage, and PROH appears more effective cryoprotectant than DMSO because PROH showed better development rates of embryos in 2 and 4-8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

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Study on Pregnancy and Viability of Frozen-Thawed Human Embryos by Cryopreservation : DMSO as Cryoprotectant (동결보존에 의한 인간배아의 생존률과 임신에 관한 연구)

  • Lee, Ho-Joon;Lee, Seung-Jae;Roh, Sung-Il;Paik, Hye-Ran;Kim, Moon-Kyoo
    • Clinical and Experimental Reproductive Medicine
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    • v.17 no.2
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    • pp.129-135
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    • 1990
  • This study was done to verify factors affecting viability after cryopreservation and pregnancy rate after frozen-thawed embryo transfer into uterus. Embryos were cryopreserved slow freezing and slow thawing and used DMSO as cryoprotectant. The results were to follows. 1. Viability of frozen-thawed embryos were 75.5% (94/105), which compared with viability of embryos according to cell stage, $2{\sim}5$ cell was 68.4% and $6{\sim}16$ cell 80.4% were significant differences (p<0.05). 2. No significant difference in duration of cryopreservation on effects affecting pregnancy rate was observed. 3. Number of embryo transfered into uterus was significant differences (p<0.05). 4. Four pregnancies resulted following replacement of 35 frozen-thawed.

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