• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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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.

• International Journal of Highway Engineering
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• v.15 no.3
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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.

• Journal of Embryo Transfer
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• v.10 no.3
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• pp.193-202
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• 1995

In order to improve the cryopreservatory techniques of livestock embryos, the quick freezing method which is directly plunged in liquid nitrogen via prefreezing procedure without freezing machine was carried out for mouse embryos treated with permeable and nonpermeable cryoprotectants. The viability of frozen-thawed embryos were evaluated by FDA vital dye test. The results obtained was summaried as follows: 1. A total of 720 embryos were recovered from frozen embryos for viability test. Evalution of the fluorescein diacetate(FDA) vital dye test with mice embryos were resulted of 2.3 total mean score - evaluted in orderly higher mean grade of P3 453 (63%), P2 133(18%), P1 51(7%) and P0 83(12%). 2. An all-round evalution of these combination, the highest viability was showed in 3M ethylene glycol + 0. 25M trehalose treated with the copper prefreezing. 3. Effects of permeable and nonpermeable cryoprotectants combination were evaluated by means FDA score. 3M ethylene glycol + 0.25M trehalose showed the highest survival rates of 2.8 mean FDA score. 4. Effects of permeable cryoprotectants were evaluated by mean FDA score but the results were not significantly different each other. 5. In evalution of the nonpermeable cryoprotectants, 0. 25M trehalose obtalned higher mean FDA score than of 0.25M sucrose and it was significantly different(P<0.05). 6. There was no significantly difference between copper and stainless-steel in prefreezing procedures.

• Journal of Veterinary Clinics
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• v.4 no.2
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• pp.449-455
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• 1987

Quick freezing of bovine embryos was attempted after they were predehydrated at room temperature. Combined solutions of 2M glycerol or 2M ethylene glycol in the presence of either 0.5 or 1.0M sucrose in phosphated buffered saline+20% calf serum were compared. The quick freezing method in which embryos were directly transferred in liquid nitrogen vapor for 2 minutes at - l70$^{\circ}C$ before being plunged into liquid nitrogen was used. Post-thaw survival rates in 2M glycerol and 2M ethylene glycol were high with 0.5 M (55.6% and 53.3%) versus 1.0M(38.1% and 31.6%) sucrose(P < 0.05). But survival rates with 2M glycerol and 2M ethylene glycol were not significantly different. Transfer thawed embryos frozen with 2M glycerol and 2M ethylene glycol by 0.5M sucrose resulted in birthrates of 40.9% and 40.0%, respectively compared to 26.3% and 27.2%, respectively, for 1.0 M sucrose(p<0.05). This was 56.0% for fresh control.

• Journal of Advanced Marine Engineering and Technology
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• v.31 no.5
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• pp.622-629
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• 2007

Some worker is occupationally exposed to cold and freezing environment. The cold stimuli in the working environment impose physiological and psychological loads on workers to decrease the task performance. The purpose of this study is to investigate the cold stimuli of cold and freezing stores widely used in Busan can make an effect on human's physiological responses and task performance, experimentally and analytically. In the experiment, 5 workers are selected as subjects, and then their skin temperatures of hand and ear, heart rates, blood pressure, and ring test performances in cold($3^{\circ}C$) and freezing($-22^{\circ}C$) stores were measured for 21 minutes and analyzed by using statistical method. It is observed that a physiological variation and the task performance are significantly influenced by an exposure time as well as a strength of cold stimuli. Also, it is suggested the exposure limiting times for the useful manual work and the performance predict model of the ring tasks. The result of this study will be useful for a fundamental data of which design the standard task time of manual tasks and solve the job placement problem of worker selection and placement in cold environment.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.3
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• pp.185-190
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• 1987

In order to study the preservation effect of $-3^{\circ}C$ partial freezing method, tile growth and biochemical activity of microorganisms and the changes of K-value in mackerel press juice were investigated at $0^{\circ}C,\;-3^{\circ}C$ supercooling (liquid phase) and $-3^{\circ}C$ freezing (solid phase). The results obtained in this paper were follows : 1) The growth and biochemical activity of microorganisms were reduced at $-3^{\circ}C$ supercooling than $0^{\circ}C$ in spite of the small variation in temperature. 2) There were no growth and biochemical activity of microorganisms at $-3^{\circ}C$ freezing (solid phase). 3) The difference in the K-value between $-3^{\circ}C$ supercooling and $-3^{\circ}C$ freezing was remarkable in spite of the same temperature.

• Journal of Embryo Transfer
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• v.30 no.3
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• pp.207-211
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• 2015

Many pronuclear stage eggs were used to generate transgenic mice (Tg) by microinjection. In this study, we used in vitro fertilized mouse eggs, followed by ultrarapid freezing to establish a simple procedure for production of Tg mice. We produced in vitro fertilized mouse eggs and cryopreserved them by ultrarapid freezing method. A total of 139 cryopreserved-thawed pronuclear eggs, of which 101 (72.6%) were survived following microinjection of chicken ${\beta}-actin$ promoter-driven firefly improved luciferase cDNA (${\beta}-act/luc^+$) and were transferred into 5 recipients. All recipients became pregnant and gave birth to a total of 15 (14.8%) pups. As a control, same DNA construction (${\beta}-act/luc^+$) was also injected into 450 in vitro fertilized eggs, of which 338 (75.1%) were survived and then were transferred into 14 recipients. Eleven (78%) mice became pregnant and littered a total of 54 (19.1%) pups. Southern blotting analysis of Tg mice indicated that one (1/15, 6.6%) and three (3/54, 5.5%) transgenic mice were production from cryopreserved and in vitro fertilized eggs, respectively. All Tg mice produced from both eggs showed the expression of improved luciferase gene. These results indicated that efficiency of produced of Tg mice from cryopreserved eggs was comparable to that from in vitro fertilized eggs. Furthermore, it is suggested that microinjection of transgene into in vitro fertilized eggs cryopreserved by ultrarapid freezing is an easy and conveniently method for production of Tg mice.

• Korean Journal of Food Science and Technology
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• v.29 no.2
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• pp.326-336
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• 1997

Four ice nucleation-active bacteria (INA-bacteria), Pseudomonas syringae, Xanthomonas campestris, Escherichia coli JM109/pEIN229 and Gluconobacter oxydans/pKIN230, were treated with heat, pressure and gamma-irradiation to compare viability and their ice nucleation activity (INA) after sterilization. Gamma-irradiated INA-bacteria showed the least decrease in T90 value (the temperature at which the 90% of drops are frozen). According to cumulative INA spectra, gamma-irradiated INA-bacteria showed little decrease in class A ice nuclei $(nucleate\;H_{2}O\;at\;higher\;than\;-5^{\circ}C)$, pressurized INA-bacteria showed more than 90% decrease in class A ice nuclei, and heat-treated INA-bacteria barely showed class A ice nuclei. Differential scanning calorimetry (DSC) was used to examine the effect of INA-bacteria on the thermophysical properties of water at freezing temperature. Freezing peaks were appeared at about $11{\sim}15^{\circ}C$ higher on thermograms and enthalpies of phase change were decreased for the water containing INA-bacteria compared with the pure water, while melting peaks were not shifted. INA measured by DSC method were significantly correlated with INA measured by drop freezing method $(R^{2}>0.993,\;p<0.0001)$, indicating that DSC can be used as a new, simple and precise method for measuring INA.

• Korean Journal of Food Science and Technology
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• v.6 no.2
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• pp.70-74
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• 1974

The round muscle of Korean cattle and squid muscle were cooked with various methods which were followed digestibility test by use of pepsin in-vitro, determination of amino nitrogen in the course of digestion procedure by using Formol method (AOAC) and influence of ether treatment for preminary test also examined. The results obtained were summarized as follows: 1. The order of digestibility values were demonstrated as follows: In case of beef, it was autoclaving, frying, raw, freezing, roasting, boiling and in case of squid muscle, it was raw, autoclaving, boiling, freezing, dry heating and roasting. 2. The amounts of amino nitrogen for beef and squid muscle were increased in proportion to digestibility value. 3. There were no significances in the digestibility between treating with ether and none of any treatment of beef and squid muscle in raw condition.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.265-271
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• 2011

This study was undertaken to find out the effect of methyl-beta-cyclodextrin (MBCD) on cold shock and membrane cholesterol quantity of sperm during the freezing process in miniature pigs. For this study, semen ejaculated from PWG M-type miniature pig was diluted that freezing solution (with egg yolk group) and m-Modena B (without egg yolk group) treated with 0, 1, 5, 10 and 20 mM MBCD before freezing process. The diluted semen was monitored sperm ability at room temperature, after cooled until $5^{\circ}C$ and after forzen-thawed for cold shock test of spermatozoa. Also, membrane cholesterol of sperm was extracted by folch solution at the same time sperm ability was assessed for viability and acrosomal status. The membrane cholesterol quantity was measured by thin-layer chromatography (TLC) method. The result, viability and acrosome integrity in semen diluted without egg yolk groups were decreased at all temperature range by increasing of MBCD concentration. In particular, sperm of egg yolk group was showed that significantly higher viability and lower acrosome damage when treated with 5 mM MBCD (p<0.05). The results of TLC experiment, cholesterol amounts were increased with MBCD cocentration in egg yolk, and decreased with MBCD concentration in m-Modena B. In cryopreservation efficiency, there was no significant difference at viability, and acrosomal state of sperm in 5 mM MBCD concentration was significantly lower in acrosome damage than other groups (p<0.05). Therefore, the addition MBCD in egg yolk was protected spermatozoa from cold shock injury. This protective effect of MBCD may be due to addition of sperm membrane cholesterol.