• Korean Journal of Animal Reproduction
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• v.8 no.2
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• pp.116-121
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• 1984

These experiments were carried out to examine the effect of rapid thawing (500$^{\circ}C$/min) on the survival of 8-cell mouse embryos cooled slowly (0.5-1$^{\circ}C$/min) to precooling temperatures between -10 and 070$^{\circ}C$ before direct transfer ofembryos to -196$^{\circ}C$, and to compare the survival of embroys thawed slowly (20$^{\circ}C$/min) and rapidly (500$^{\circ}C$/min) after in vitro culture. In addition, the sensitivity of 8-cell mouse embroys to the rate of addition and removal of cryoprotectant at room temperauture, and the effect of developing stages on the survival of embryos frozen-thawed slowly were investigated. The results obtained were summarized as follows: 1. Embryos were survived in rapid thawing (500$^{\circ}C$/min) only when slow cooling was terminated at relatively high subzero teperaure (-20 to -60$^{\circ}C$). The highest survival rate(77.0%) in in vitro culture of embryos thawed rapidly was obtaeined after transfer to -196$^{\circ}C$ from -40$^{\circ}C$. 2. For the survival of embryos in slow thawing (20$^{\circ}C$/min.), slow cooling to lower subzero temperature (-50$^{\circ}C$ and below) was required before transfer of embryos to -196$^{\circ}C$. These results indicate that embryos transferred to -196$^{\circ}C$ from high subzero temprature contain much interacellular ice to damage them during slow warming but to permit survival of embryos after rapid warming. 3. The Survival rate (72.7%) of 8-cell mouse embryos after rapid addition and removal of cryoprotectant, DMSO at room temperature was similar to that (83.9%) after slow addition and removal of cryoprotectant at same temperature. 4. The survival rates of 1-, 2-, 4- and 8-cell embryos frozen-thawed slowly were 26.7, 76.4, 70.0 and 83.9%, respectively.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.25 no.2
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• pp.79-92
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• 1992

This study was carried out to develop new techniques useful for cryopreservation, thawing and artificial insemination, and ultrastructural changes of cryopreserved spermatozoa in rainbow trout(Oncorhynchus mykiss) . Two extenders, such as Tyrode solution and Whittingham's $T_6$ solution, were used to preserve rainbow trout sperm in refrigerator $(-20,\;-40\;and\;-70^{\circ}C)$ or liquid nitrogen $%(-196^{\circ})$. Hand-stripped semen was diluted to 1:16 with two extenders, an then the semen were frozen after mixing semen and each extender containing 1M or 1.5M DMSO solution to 1:1. After 60 days cryopreserved semen was thawed in a $13^{\circ}$ water bath, and subsequently centrifugated. After centrifugation at 1,000 rpm for 5 min thawed semen was washed with extenders, and then fertilized with fresh eggs. The results obtained in these experiments were summarized as follows: After cryopreservation, over 75% of spermatozoa were appeared motile and the survival rate was high. Following cryopreservation by the addition of cryoprotectant such as DMSO, methanol and glycerol, the fertilization rate of the thawed spermatozoa appeared over $99\%$ compared with the control having $99\%$ of fertilization rate. There was no difference between the control and experimental groups such as $(-20^{\circ}C\;-40^{\circ}C\;and\;-70^{\circ}C)$ and $-196^{\circ}$ in fertilization rate. Following cryopreservation at $-196^{\circ}$ by the addition of 1M DMSO of cryoprotectant, each fertilization rate following 24 hours and hatching rate following 24 days showed $96\%$ and $8\%$ by the addition of BSA, but showed $98\%\;and\;10%$ by no addition of BSA. Following 2 months of cryopreservation by the addition of 1M DMSO of cryoprotectant, there were $10%$ of hatching rate at $-196^{\circ}\;and\;10\%\;and\;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1M methanol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C,\;and\;28\%,\;at\;-70^{\circ}C$ Following 2 months of cryopreservation by the addition of 1M glycerol of cryoprotectant, there were $22\%$ of fertilization rate at $-20^{\circ}C$, and $33\%,\;at\;-70^{\circ}C$. pollowing 2 months of cryopreservation by the addition of 1.5M DMSO of cryoprotectant, there were $27\%$ of fertilization rate at $-20^{\circ}C,\;an\;36\%\;and \;35\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C$. Following 2 months of cryopreservation by the addition of 1.5M glycerol of cryoprotectant, there were $34\% \;of\;fertilization\;rate\;at\;-20^{\circ}C, \;and\;31\%\;and\;31\%,\;respectively,\;at \;-40^{\circ}C\;and\;-70^{\circ}$. Following 2 months of cryopreservation by the addition of 1.5M methanol of cryoprotectant, there were $28\%$ of fertilization rate at $-20^{\circ}C,\;and\;29\%\;and\;28\%,\;respectively,\;at\;-40^{\circ}C\;and\;-70^{\circ}C.$ From 10 days and 15 days following fertilization at $13^{\circ}C\;and\;10^{\circ}C$, respectively, the mortality rate of fertilized ova was markedly increased. The middle piece of spermatozoa had two set of central doublets, nine set of outer coarse fibres, and mitochondrial sheath. Spermatozoa went through morphological changes during storage, e.g. winding of flagella, detachment of the nuclear envelope and the plasma membrane from the nucleus of the sperm head. There were $1\%$ abnormal spermatozoa in fresh sperm and about $15\%$ during storage.

• Journal of the Korean Society for Heat Treatment
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• v.15 no.4
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• pp.166-171
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• 2002

The effects of Ni and N addition to 304 stainless steel on mechanical properties and microstructure were investigated within temperature ranged from $-196^{\circ}C$ to $25^{\circ}C$. The addition of nickel and nitrogen induced the stability against the formation of provide significantly enhanced ductility at $-196^{\circ}C$ and reduce the Md temperature. Fe-18Cr-10Ni-0.2N alloy showed the ${\gamma}$ phase in deformed structure which it was beneficial to enhance the tensile strength and elongation at $25^{\circ}C$ and $-196^{\circ}C$.

• Korean journal of food and cookery science
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• v.13 no.2
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• pp.106-112
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• 1997

The effect of low concentrations of allspice (Pimenta dioica L.) in culture broth as an antibacterial agent against Escherichia coli 0157:H7 and Staphylococcus aureus 196E was tested at 35,5 and -20$^{\circ}C$. Tryptic soy broth (TSB) containing 0∼2% (w/v) of allspice was inoculated with 10$\^$5/∼10$\^$6/ cells/$m\ell$ of E. coli and S. aureus and incubated at each temperature. The growth of E. coli was not inhibited at 0.1∼1.0% allspice and growth occured at 2％ allspice but only after a prolonged lag period. Growth of S. aureus was inhibited with increasing concentration of allspice at 35$^{\circ}C$. Growth of S. aureus occured at the presence of 0.1∼0.3% allspice but the viability of S. aureus at 0.5∼2.0% allspice was decreased during storage at 35$^{\circ}C$. During refrigerated storage at 5$^{\circ}C$, inhibition of E. coli and S. aureus was increased with the progress of time and increasing spice concentration. During frozen storage at -20$^{\circ}C$, antibacterial activity of allspice against E. coli was increased with increasing storage time and spice concentration while that activity against S. aureus was effective during early period of storage. There was no major changes in population of S. aureus in TSB with different concentration of spice frozen at -20$^{\circ}C$. Viable counts of E. coli and S. aureus at 0.l％ of allspice was less than that of control during frozen storage.

• Journal of the Korean institute of surface engineering
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• v.37 no.1
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• pp.64-70
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• 2004

Cryogenic characteristics of austenitic stainless steel based on 304 steel with nickel and nitrogen were investigated at room temperature and $-196^{\circ}C$. The alloys were fabricated by vacuum arc furnace and cold working after homogenization treatment. The addition of nickel and nitrogen decreased the stability of $\delta$-ferrite and induced the stability against the formation of martensite to result significantly in enhancing ductility at $-196^{\circ}C$. Nitrogen reduced Md temperature, which was beneficial to the tensile strength and elongation at $25^{\circ}C$ and -196$^{\circ}C$.

• Korean Journal of Animal Reproduction
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• v.8 no.1
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• pp.22-28
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• 1984

This experiment was carried out to investigate effects of DMSO or ethylene glycol as a cryopotectant and of freezing methods on survival rate of forozen-thawed rat 2-cell embryos by morphological observation. 2-cell embryos were recovered from oviducts of Sprague Dawley females mated with males of same strain on day 2 of pregnancy after inducing superovulation by intrapertioneal injection of PMSG and HCG. In slow freezing and thawing groups, embryos were frozen to -79$^{\circ}C$ or -196$^{\circ}C$ in a glass test tube containing 0.2ml PBI with 1.5M DMSO or 1.2M ethylene glycol at a rate of 0.3-1.0C/min. and thawed slowly. When samples were frozen to -79$^{\circ}C$, higher survival rate was obtained in the medium containing DMSO (43.9%) than ethylene glycol (41%). And similar result was obtained (32.5% in DMSO vs. 31.4% in ethylene glycol) when samples were frozen. In rapid freezing and thawing groups, embryos were frozen to -79 or -196$^{\circ}C$ in a glass test tube containing 0.2ml of PBI with 1.5M DMSO or 1.2M ethylene glycol by rapid cooling, and thawed rapidly. When samples were frozen to -79$^{\circ}C$, 1.5M DMSO (13.2%) was more effective than 1.2M ethylene glycol (6.1%). When the storage temperature was -196$^{\circ}C$, survival rates were 9.8% in 1.5M and 5.4% in 1.2M ethylene glycol.

• Food Science and Preservation
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• v.22 no.1
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• pp.44-50
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• 2015

This study was performed to optimize the preparation of frozen lotus roots. Prior to freezing, an optimal blanching condition at $100^{\circ}C$ for 5 min was established, based on the microbial growth, texture, total phenolic content (TPC), and sensory evaluation results. The blanched samples were then frozen under various freezing conditions ($-20^{\circ}C$ in a freezer for 2 hr, $-70^{\circ}C$ in a gas nitrogen convection chamber for 7 min, and $-196^{\circ}C$ in liquid nitrogen for 20 sec), and their qualities after thawing were determined. The scanning electron microscopic analysis indicated that the microstructure of the sample frozen at $-70^{\circ}C$ was similar to that of the control sample, compared with the other freezing conditions (-20 and $-196^{\circ}C$). The antioxidant activities of the frozen samples decreased compared to those of the control, but there was no significant (p<0.05) difference among the treatments. In terms of TPC, the samples frozen at -70 and $-196^{\circ}C$ had significantly (p<0.05) higher values than the sample frozen at $-20^{\circ}C$. In addition, the drip loss of the sample frozen at $-20^{\circ}C$ was higher than those of the other frozen samples. These results suggest that freezing at $-70^{\circ}C$ in a gas nitrogen convection chamber can be an optimal freezing method of producing high-quality frozen lotus roots.

• Journal of Applied Biological Chemistry
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• v.58 no.1
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• pp.55-60
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• 2015

To secure the microbial safety of frozen strawberries, they were treated with the combined solution of aqueous chlorine dioxide and acetic acid prior to freezing and the effects of different freezing methods (at $-20^{\circ}C$ in a freezer, at $-70^{\circ}C$ in a gas nitrogen convection chamber, and at $-196^{\circ}C$ in liquid nitrogen) on the quality changes of strawberries were examined. Regarding the color of frozen strawberries, there were negligible changes among freezing treatments. In contrast, vitamin C content and sensory evaluation scores of strawberries frozen at $-70^{\circ}C$ were the highest among the samples. Drip loss of strawberries frozen at $-70^{\circ}C$ was the lowest as 14.39%, compared with strawberries frozen at -20 and $-196^{\circ}C$. In addition, the effects of combined treatment of 50 ppm chlorine dioxide and 1% acetic acid on the microbial growth in frozen strawberries were investigated, and the populations of preexisting microorganisms in the frozen strawberries were not detected by the combined pre-treatment. These results suggest that rapid freezing at $-70^{\circ}C$ using a gas nitrogen convection chamber is an appropriate freezing method for preserving quality of strawberries, and as a pre-freezing treatment, the combined treatment of aqueous chlorine dioxide and acetic acid can be effective for improving microbiological safety of frozen strawberries.

• Proceedings of the Korean Society of Marine Engineers Conference
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• 2002.05a
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• pp.113-199
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• 2002

In this study, the toughness was evaluated by using the instrumented Charpy impact testing procedures for A15083-O aluminum alloy used in the LNG carrying and storing tank. The specimens were GMAW welded with four different mixing shield gas ratios (Ar100%+He0%, Ar67%+He33%, Ar50%+He50%, and Ar33%+He67%), and tested at four different temperatures(+25, -30, -85, and -196$^{\circ}C$ ) in order to investigate the influence of the mixing shield gas ratio and the low temperature. The specimens were divided into base metal, weld metal, fusion line, and HAZ specimen according to the worked notch position. From experiment, the maximum load increased a little up to -85$^{\circ}C$, and the maximum load and maximum displacement were shown the highest and the lowest at -196$^{\circ}C$ than the other test temperatures. The absorption energy of weld metal notched specimens was not nearly depends on test temperature and mixing shield gas ratio because the casting structure was formed in weld metal zone. In the other hand, the others specimens was shown that the lower temperature, the higher absorption energy slightly up to -85$^{\circ}C$ but the energy was decreased so mush at -196$^{\circ}C$

• Journal of Advanced Marine Engineering and Technology
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• v.26 no.6
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• pp.653-660
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• 2002

In this study, the toughness was evaluated by using the instrumented Charpy impact testing procedures for A15083-O aluminum alloy used in the LNG carrying and storing tank. The specimens were GMAW welded with four different mixing shield gas ratios (Ar100%+He0%, Ar67%+He33%, Ar50%+He50%, and Ar33%+He67%), and tested at four different temperatures(+25, -30, -85, and $-196^{\circ}C$) in order to investigate the influence of the mixing shield gas ratio and the low temperature. The specimens were divided into base metal, weld metal, fusion line, and HAZ specimen according to the worked notch position. From experiment, the maximum load increased a little up to -$85^{\circ}C$ , and the maximum load and maximum displacement were shown the highest and the lowest at -$196^{\circ}C$ than the other test temperatures. The absorption energy of weld metal notched specimens was not nearly depends on test temperature and mixing shield gas ratio because the casting structure was formed in weld metal zone. In the other hand, the other specimens were shown that the lower temperature, the higher absorption energy slightly up to $-85^{\circ}C$ but the energy was decreased so mush at $-196^{\circ}C$.