• International Journal of Highway Engineering
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• v.12 no.3
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• pp.71-77
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• 2010

Anti-freezing layer does not used in case of non frost heaving in subgrade soils. In this case, the modulus of subgrade soils were varied with freezing and thaw cycles under non frost heaving. That effect should be properly considered in pavement design. Impact resonance test that is nondestructive testing method was used for continuously determining the modulus of subgrade soils during freezing and thaw cycle. The modulus of subgrade soils was identical with freezing and thaw cycles under closed freezing and thaw system which is no water supplement into specimen during testing. There was also no difference in the modulus of subgrade soil between before and after freezing-thaw cycles for all specimens with different water content and density. That is thaw-weakening of subgrade soils do not occur under closed freezing and thaw system. The moduli at freezing conditions are varied with water content and density, but it can be ignored in practical design sense.

• International Journal of Industrial Entomology and Biomaterials
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• v.47 no.2
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• pp.126-133
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• 2023

Genetic resources of mulberry trees are commonly preserved as trophosomes, which are vulnerable to environmental factors, such as natural disasters, diseases, and pests. This study establishes a basic protocol for ultra-low temperature cryopreservation of mulberry trees using a two-step freezing process. The procedure was established using the "Daeshim" variety and then tested on genetic resources from 24 other mulberry varieties. Samples were first dried to a moisture content of 33-43% in a low-temperature forced-air chamber at -5 ℃, then slowly frozen from -5 ℃ to -20 ℃, and preserved in liquid nitrogen (-196 ℃). To determine the regeneration rate, isolated dormant buds were inoculated into MS basal medium, and grown shoots were grafted onto 1-year-old rootstock via chip budding and then cultured. After freezing in liquid nitrogen, the "Daeshim" variety exhibited a survival and regeneration rate of more than 70% and 50%, respectively. Applying the two-step freezing process to genetic resources from 24 mulberry species yielded average survival and regeneration rates of 85.3% and 75.5%, respectively. Morus alba showed survival and regeneration rates of 100%, confirming the efficacy of the two-step freezing method. These results indicate the high feasibility of ultra-low-temperature cryopreservation through two-step freezing of dormant buds from mulberry genetic resources. Additional research is required into the variations in regeneration rates with freezing period in liquid nitrogen.

• Ocean and Polar Research
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• v.33 no.3
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• pp.303-308
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• 2011

Polar psychrophiles which thrive under extreme conditions such as cold temperature, high salinity, and high dose ultraviolet light, emerge as novel targets for biotechnology. To prevent genetic drift and the possibility of contamination by subculturing, cryopreservation was employed for two psychrophilic microalgae, Porosira sp. (KOPRI AnM0008) and Pyramimonas sp. (KOPRI AnM0046), which have anti-freeze activities. Five cryoprotectants (dimethyl sulphoxide, ethylene glycol, glycerol, methanol and propylene glycol) showed toxicity at 20-30% (v/v). The optimal cryoprotectant concentration and equilibration time were less than 20% and 10 min, respectively. Cryopreservation was carried out in the presence of cryoprotectants either by direct freezing in liquid nitrogen ($LN_2$) or controlled freezing using a controlled rate freezer followed by storage in the $LN_2$ tank. As a result, Pyramimonas sp. (KOPRI AnM0046), a psychrophilic chlorophyta was revived. Cryopreserved Porosira sp. was not revived from either freezing protocols probably due to the silicic cell wall and its relatively large cell size. In the case of Pyramimonas sp. (KOPRI AnM0046), the controlled freezing method showed higher revival yield than the direct freezing method.

• Journal of Biosystems Engineering
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• v.33 no.3
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• pp.167-172
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• 2008

The objective of this study was to investigate the adequate storage method that was able to maintain the high quality of rough rice according to storage methods and period. The quality change of the rough rice during the storage period was evaluated by storage method such as cooling bin using winter cold air, ordinary temperature bin, freezing, refrigeration and indoor storage. Moisture content, brown rice whiteness, hardness, crack ratio and germination ratio were measured in this study. Moisture content of rough rice stored in cooling bin using winter cold air and ordinary temperature bin were decreased by 0.07% and 0.42%, respectively, which were lower than the other storage method. The hardness of brown rice increased in order of storage method such as winter cooling bin, normal bin, freezing storage, refrigeration storage and indoor storage. Crack ratio by indoor and ordinary temperature bin storage were increased by 2.68% and 3.63%, respectively, whereas cooling bin using winter cold air, refrigeration and freezing storage showed below 1.0%. The highest germination rate was found in cooling bin using winter cold air. As a result, cooling bin using winter cold air can be evaluated for the adequate storage method of rough rice.

• ALGAE
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• v.21 no.4
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• pp.479-483
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• 2006

In preparation of the biological samples for electron microscopy, the chemical fixation by glutaraldehyde, paraformaldehyde, and OsO4 has been generally used for a long time. However, the chemical fixation method has some problems: the infiltration time is a little bit long and the ultrastructure of cell or tissue transforms before complete fixation of sample. So, recently, cryo-fixation is considered more often in biomedical field. In this study, we compared High Pressure Freezing (HPF) method with chemical fixation method using a algal sample (Ptilota filicina J. Agardh), which was difficult to fix using chemical fixation method. In chloroplast, the ultrastructure of thylakoid lamella and phycobilisome can not show clearly by chemical fixation. In this study we could observe the ultrastructure of thylakoid lamella and phycobilisome of chloroplast very clearly using HPF fixation. An improved images of ultrastructures of nucleus, mitochondrion and floridean starch could obtain. These results suggest that HPF method is very useful method in algal specimen for electron microscopy.

• Journal of Korean Tunnelling and Underground Space Association
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• v.19 no.6
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• pp.1059-1075
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• 2017

The objective of this study was to evaluate the freezing and thawing resistance of the existing drainage system for leakage treatment of underground concrete structures operating in cold regions. The freezing and thawing test was conducted on 4 types of drainage system specimens to evaluate the freezing and thawing resistance of the drainage system. The freezing and thawing resistance was evaluated on 4 types of Hotty-gel, as a waterproofing material, connection methods and on two methods to fix the drainage board with Hotty-gel on the surface of cement concrete specimen. One cycle of the freeze-thaw testing was 48 hours (24 hours of freezing and 24 hours of thawing), and the temperatures of freezing and thawing were at $-18^{\circ}C$ and $10^{\circ}C$, respectively. Among the 4 types of Hotty-gel connection methods, leakage occurred after 28 cycles (8 weeks) of freeze-thawing only in the Hotty-gel connection method with the 'V' groove applied to the corner of the drainage board. No leakage occurred in the 3 types of Hotty-gel connection methods. In two fixing methods, leakage occurred in the method of fixing the drainage board on the cement concrete specimen using the washer, screw and plastic wall plug. Leakage occurred at one point after 10 cycles (3 weeks) of freezing and thawing. After 28 cycles (8 weeks) of freezing and thawing, leakage point increased to 5 points. As time passed, the leak point was not increased, but the amount of leakage was increased at each leak point. The Hotty-gel connection method with cross-sectional diagonal shape was evaluated to be the highest in the production efficiency considering the production time and manufacturing method of the Hotty-gel connection shape. In the construction efficiency considering the construction time and construction method, the fixing method of air nailer, fixed nail and washer was superior to that of the washer, screw and plastic wall plug.

• Proceedings of the Korea Concrete Institute Conference
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• 1997.04a
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• pp.155-161
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• 1997

This Expriment is performed to describe the properties of the freezing-thawing and to find the method to enhance the freezing-thawing resistance of the high strength light weight aggregates concrete. For this purpose, we made 8 kinds of specimen of concrete mold. The light weight coarse aggregate concrete which contained AE was appeared in good condition and its durability index was more than 90% by the buffer action which owing to entained air. The light weight aggregates concrete which admixture of silca fume, was appeared that the durability index was 46.74% in spite of its high strength. I might conclude that the most important factor for freezing-thawing resistance of high strength light weight aggregate concrete is the entrained air.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.2
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• pp.67-72
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• 2005

Overwintering capacity, closely related to winter hardiness, of Chinese milk vetch planted with different sowing times and sowing practices was investigated to improve the incorporation into cropping system in Korea. The tolerance to low temperature was evaluated with $LT_50$ using leaf disc leaching method. Dry weight of CMV was reduced remarkably with delayed planting from Sep. 5 to Oct. 20. The differences in tolerance to freezing temperature were not conspicuous among CMV genotypes, however, the differences between genotype (collections at different regions) were due to the plant architecture, mainly to the leaf angle. The crouching genotype collected at central region of Korean peninsula, which showed excellent freezing tolerant, has planophile leaves. The feature of internal constituents of CMV genotypes did not show any noticeable differences with respect to the freezing tolerance which evaluated by leaf disc leaching experiment. To overcome the poor overwintering capacity, tolerant genotype should be developed by selection with considering the plant architecture. The reduction of CMV growth during overwintering period was ameliorated with furrow-sowing under late-sown condition, therefore, when the CMV is inevitably sown late after recommended time, the seeds should be sown on furrow to overcome the cold stress.

• Journal of Embryo Transfer
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• v.7 no.1
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• pp.13-19
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• 1992

This study were carried out to investigate the effective concentration of cryoprotective agents and sucrose by one-step straw method, and to determine the optimum thawing temperature and equilibration time of frozen porcine embryos. The porcine embryos foflowing dehydration by cryoprotective agents and a various concentration of sucrose were directly plunged into liquid nitrogen and thawed in 3$0^{\circ}C$ water. Survival rate was defined by FDA test. The results are sunnnarized as follows : 1. The survival rates of porcine embryos after ultrarapid frozen4hawing in the freezing medium with a various concentration of glycerol, DMSO and propanediol added 0.25M sucrose were higher survival rate than those of sucrose concentration of 0.50M. 2. The survival rates of porcine embryos after ultrarapid ftozen4hawing in the freezing medium added 0.25M and 0.SOM sucrose were higher survival rate than those of sucrose concentration of 0.75M and 1.00M. 3. The temperature thawed at 2$0^{\circ}C$ and 3$0^{\circ}C$ resulted in a significantly higher embryos survival rate after 72 hrs in culture than did at 35$^{\circ}C$. 4. The equilibration time on the survival rate of porcine embryos was attained after short period of time(2.5~5 min.) in the freezing medium higher than long period of time(10~20 min.).

• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.191-198
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• 2001

Objective : This study was conducted to examine the effect of vitrification on the survival and in vitro development of mice 1-cell zygotes. Method: Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. The 1-cell zygotes were also subjected to a slow freezing-thawing method to compare with vitrification method. Solution composed of ethylene glycol (6.0 M, 5.0 M, 4.0 M) and sucrose (1.0 M) were used as cryopropectant. The experiments employed the method loading the embryos on electron microscope grids. Results: I. The effects of exposure in vitrification solution. 1-cell zygotes were non-toxic at all concentrations of the vitrification solution showing the survival rate between 88.1% and 97.5%. Development into 2-cell was more successful in the higher concentrations of the vitrification solution. Therefore, higher concentrations of the vitirification solution do not seem to cause any problems in vitrification procedure. II. The effects of vitrification method. 1-cell zygotes showed the survival rate between 78.8% and 92.4%. The lowest and the highest survival rate was observed in the 6.0 M and 4.0 M vitrification solution, respectively. 2-cell development rates varied from 77.6% to 91.3%. Blastocyst development rate was shown highest in 5.0 M and the lowest in 4.0 M solution. Therefore, the highest 2-cell and blastocyst development rate was observed in 5.0 M solution. III. Comparison of vitrification and slow freezing-thawing method on 1-cell zygotes. This experiment showed that 1-cell zygotes had the highest survival and development rates in 5.0 M vitrification solution. Vitrified group of 1-cell zygotes, in the 5.0 M vitrification solution, were compared with the group processed in slow freezing-thawing method. The development rate into 2-cell and blastocyst as well as the survival rate were higher in the vitrified group than in the slowly freezed group. Conclusion: 1. The results demonstrate that the best cryoprotectant is a 5.0 M vitrification solution for 1-cell zygotes. 2. Vitrification method significantly increases the survival rate of the 1-cell zygote and its development into 2-cell and blastocyst. Equilibration and exposure time during the vitrification was remarkerbly short in this experiment. Total time, from the exposure to vitirification solution to storage in the liquid nitrogen, was taken only 90 seconds. In contrast, the slow freezing-thawing method have taken more than four hours. Taken together, we presume that the overall time used for the procedure contributes to the results as an important parameter. 3. The loading of 1-cell zygotes on the EM grid is technically more simple and takes less time than the straw or cryo vial method.