• Development and Reproduction
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• v.4 no.2
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• pp.161-173
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• 2000

The aim of this study was to assess the effect of the frequency of the L$N_2$ infusion on the ultrastructure, metabolism and development of the embryo after freezing and thawing by computerized cell freezer. Two-cell embryos of ICR mouse were randomly allocated into fresh (control), high-frequency freezing (group 1) and low-frequency freezing (group 2). For fresh and frozen-thawed intact 2-cell embryos, total ceil number in the blastocyst was counted by fluorescent microscope after Hoechst 33258 staining. Relative amount of $H_2O$$_2$ was measured by DCHFDA. Intracellular location and membrane potential of the mitochondria were evaluated by staining with rhodamine 123 and JC-1. The structure of actin filament was also evaluated by confocal microscope. DNA fragmentation was assessed by TUNEL method after development into blastocyst. The survival rate of intact embryo was higher in group 1 than group 2 (50.7％ vs. 34.6％ respectively, p<0.05). The blastocyst developmental rate was significantly low in group 2 (86.7％, 76.7％ vs. 44.0％ for control, group 1 and group 2 respectively, p<0.05). Total cell number in the blastocyst was also significantly lower in group 2 than control (79.5$\pm$12.9, 71.6$\pm$8.0, and 62.5$\pm$4.7 for control, group 1 and group 2 respectively, p<0.05). The relative amount of $H_2O$$_2$ was higher in group 2 than other groups (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). After JC-1 staining, relative intensity of mitochondria with high membrane potential was significantly lower in group 2 than control and group 1 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). In group 2, partial deletion and aggregation of the actin filament was found. DNA fragmentation rate was also hieher for group 2 versus other groups (30.8％, 36.0％ vs. 65.6％, p<0.05). The frequency of the L$N_2$ infusion is an important factor for the development of frozen-thawed mouse embryo. High-frequency infusion may prevent damages of cytoskeleton and mitochondria in the embryo probably by preventing the temperature fluctuation during dehydration phase. We speculate that the application of high-frequency infusion method in human embryo may be promising.

• Journal of The Geomorphological Association of Korea
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• v.20 no.3
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• pp.95-107
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• 2013

In order to examine the temperature regime responsible for periglacial processes, air and ground temperatures were monitored from October 2010 to May 2011 at a subalpine bare patch (1,710m asl) of Mt. Halla. Four thermistor sensor probes were installed at 55 cm above a ground surface and depths of 2 cm, 10 cm, and 20 cm, respectively. A mean air temperature is $-0.1^{\circ}C$, while mean ground temperatures are $1.8^{\circ}C$ at 2 cm, $2.6^{\circ}C$ at 10 cm and $3.2^{\circ}C$ at 20 cm deep. A mean monthly ground temperature at 2 cm deep demonstrates below $0^{\circ}C$ successively from January to March, while those at 10 cm and 20 cm deep show no sub-zero temperature. A total of 72 freeze-thaw cycle was observed in air temperature. However, the numbers in ground temperature reduced into 17 at 2 cm, 8 at 10 cm, and 3 at 20 cm deep. The cycles of air temperature and ground temperature at 2 cm deep mostly fluctuated diurnally, while those of ground temperature at 10 cm and 20 cm deep exhibited a several-daily oscillation. Snow cover over 55 cm high remained from January to early April, and it seemed to disappear completely on April 16. A seasonal frost of at least 2 cm thick was formed on late December and the isotherm of $0^{\circ}C$ descended slowly into 10 cm deep on late March to early April due to the insulating snow cover. It showed the maximum freezing depth of 20 cm on April 7 to 14 and then thawed rapidly so that the frozen ground did not longer after April 17. Periglacial processes are predominant during a freezing period than a thawing period when the ground surface is still covered with snow. The periglacial mass movement in the subalpine zone of Mt. Halla is mainly generated by frost creep in terms of the occurrence depth of diurnal freeze-thaw cycle and the maximum freezing depth of ground.

• Food Science and Preservation
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• v.18 no.5
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• pp.661-668
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• 2011

This study was conducted to devise appropriate blanching-process conditions as a means to convert Doraji, which is widely used in Korean food due to its unique fragrance and flavor, into frozen food materials for various uses. For the Hunter L values representing the brightness transformation among the surface color and gloss changes that were observed in Doraji before and after freezing, and after Doraji went through a blanching process, the specimen that went through a blanching process at $80^{\circ}C$ showed a significantly higher value compared to another specimen processed at a higher temperature, and the first specimen's value also rose after freezing. Meanwhile, for the hardness values, they declined more as the blanching temperature became higher and as the processing time became longer. For the number of total counts and the number of coliform groups, the number of total counts at $3.75{\times}10^5$ and $1.25{\times}10^5$ cfu/g before the blanching process was reduced into the approximately 2-3 log scale, and no coliform group was detected after the blanching process. As for the peroxidase activity, its activation was decreased by the blanching process, and more than 89% of the peroxidase became inactivated in all the specimens that went through the blanching process. The sensory characteristics of the frozen-thawed Doraji by test group showed the radish leaves blanched at $90^{\circ}C$ for 1 min to be the most highly evaluated in terms of the overall preference level (p<0.05).

• Korean Journal of Environmental Agriculture
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• v.36 no.4
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• pp.256-262
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• 2017

BACKGROUND: Seed of perilla (Perilla frutescens var. japonica Hara) is short-lived in conventional storage conditions. For long-term conservation of plant species, cryopreservation is the method currently available. This study was performed to find out reliable methods for a long-term storage of seeds of perilla as a genetic resource. METHODS AND RESULTS: Using seeds of 9 perilla cultivars, the effects of desiccation, aging, and cryopreservation on seed germinability and ascorbate peroxidase activity in the seeds were investigated. Initial germinability of the seeds was various, and dry seeds of all cultivars survived cryopreservation without loss of viability. The highest germination was achieved at 4-5% moisture content, and stimulatory effect of cryogenic temperature on the seed germination was observed in some cultivars. Accelerated aging of perilla seeds led to reduction in germination and ascorbate peroxidase activity, and the susceptibility of seeds to aging was different among the tested cultivars. No significant difference in germination was observed for the aged seeds of control and liquid nitrogen exposed. CONCLUSION: The results of this study suggest that cryopreservation at 4-5% moisture content would be a suitable method for long-term conservation of perilla seeds without detrimental effects on germination.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.241-248
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• 2014

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). Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by PMSG and hCG. Two-step EG, DMSO and 4-step EG, 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. As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in EG and DMSO was significantly higher than 4~8 cell (65.4% versus 61.2%, 81.1% versus 72.5%) (p<0.01, p<0.01), but the development rates of 4~8 cell embryos in EG 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 EG 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 EG(77.0% versus 64.4%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The development rate from morular to hatching blastocyst, however, was sinificantly higher in EG than in DMSO during 48 hr (p<0.01). The survival rate of 4~8 cell embryo was 62.5% in EG and 73.3% in DMSO. The development rates of embryo in EG 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. The survival rate of embryo in 2 cell stage was higher than in 4~8 cell stage, and EG appears more effective cryoprotectant than DMSO because EG 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.

• Korean Journal of Animal Reproduction
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• v.18 no.1
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• pp.31-37
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• 1994

This study carried out to investigate the effects of cryoprotectants in the medium on the survival rate of rapidly frozen porcine bisected demi-embryos. The porcine bisected demi-embryos following dehydration by cryoprotectants containing sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water bath. Survival rate was defined as development rate on in vitro culture or FDA-test. The results are summarized as follows : 1. The survival rates of without-zona pellucida embryos and 2 blastomeres porcine embryos were 10.0 and 7.1%, respectively. The rate of unfrozen blastomeres (20.00%) was significantly higher than that of non-frozen oocytes. 2. The survival rates of with and without-zona pellucida of bisected porcine embryos by micromanipulator were 20.0 and 14.3%, respectively. 3. The developmental rates of with and without-zona pellucida of bisected poricine embryos by micromanipulator were 13.3 and 7.1%, respectively.

• Korean Journal of Animal Reproduction
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• v.18 no.1
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• pp.25-29
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• 1994

This study was carried out to investigate the in-vitro fertilization rate survival rate of rapidly frozen porcine immature embryos. The porcine embryos following dehydration by cryoprotectants containing sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water bath. Survival rate was defined as development rate on vitro culture or FDA test. The results are summarized as follows : 1. The in vitro fertilization rate of all frozen immature oocytes(6.7~26.7%) was very low, 35.0% of unfrozen oocytes and the rate of immature oocytes was very higher than that of mature oocytes. 2. The survival rate of all frozen immature oocytes(10.3~25.0%, 13.3~30.0%) was very low, 45% of unfrozen oocytes and the rate of immature oocytes was slightly higher than that of mature oocytes.

• Journal of Embryo Transfer
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• v.4 no.1
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• pp.35-40
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• 1989

The effects of cryoprotectants (glycerol, DMSO and ethylene glycol) and the concentrations (0, 0, 25, 0.5and 1.0 M) of sucrose in the diluent on the is vitro survival of mouse morulae froaen rapidly in liquid nitrogenvapour were examined. When the embryos were equilibrated in 1.5 M cryoprotectants +0.25 M sucrose in one-step or in 3.0 M cryoprotectants +0.25 sucrose in two-step and diluted with 0, 0.25, 0.5, or 1.0 M sucrose solution after thawing, high survival rates were obtained in ethylene glycol (48.0% to 88.2 %) or in glycerol (35.0 % to 77.8 %). These results show that 1.5 M ethylene glycol is a highly efficient cryoprotective agent for the rapid freezing of mouse morula embryos and 0.5 M sucrose was optimal concentration in the diluent after thawing.

• 축산식품과학과 산업
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• v.5 no.2
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• pp.58-69
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• 2016

건강지향의 식육, 육제품에 대한 사회적 요구에 부응하기 위해 기능성 첨가물 대체 연구로부터 원재료인 신선육과 가공공정 중 식육 단백질의 변성을 제어하고 최적화하기 위한 연구과제로써 온도체 가공/급속동결송풍냉각/저온 세절혼합 조합 기술(처리구)에 대한 유효성이 평가되었다. 그 결과 새로 제안된 조합 공정기술은 기존의 냉도체/수침냉각/세절혼합 조합(대조구)과 비교했을 때 단백질 변성 제어를 통한 기능성 단백질 추출성의 향상으로 인해 신선육의 품질향상뿐만 아니라 가열 처리 후 단백질 겔의 수율과 조직감 모두 증진되었으며, 특히 나트륨을 저감화한 조건($2%{\rightarrow}1%$)에서도 기존의 대조구와 유사한 특성을 보여 나트륨 저감화를 위한 기술로 제안될 수 있었다.

• Food Science and Preservation
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• v.24 no.2
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• pp.168-180
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• 2017

This study was performed to determine the rapid thawing method for reducing the thawing time of frozen pork loins and to examine the effects of supercooling on the microbiological, physicochemical, and sensory qualities of fresh and frozen-thawed pork during storage at -1.5, 4, and $15^{\circ}C$. Forced-air thawing at $4^{\circ}C$ was the most time-consuming process, whereas radio frequency thawing time was the shortest by dielectric heating. The supercooling storage temperature was chosen to be $-1.5^{\circ}C$ because microstructural damages were not observed in the pork sample after cooling at $-1.5^{\circ}C$ for 24 h. Fresh or frozen-thawed pork loins stored at $-1.5^{\circ}C$ had lower drip loss and total volatile base nitrogen, thiobarbituric acid-reactive substance, and Hunter b* levels than loins stored at 4 and $15^{\circ}C$. In addition, the least degree of increase in preexisting microorganisms counts of the fresh or frozen-thawed pork loin samples was obtained during supercooled storage at $-1.5^{\circ}C$. Sensory quality results of fresh and frozen-thawed pork loin samples stored at $-1.5^{\circ}C$ showed higher scores than the samples stored at 4 and $15^{\circ}C$. These data indicate that supercooling at $-1.5^{\circ}C$ in the meat processing industry would be effective for maintaining the quality of pork meats without ice crystal nucleation and formation.