• Journal of Embryo Transfer
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• v.13 no.2
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• pp.173-178
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• 1998

The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% $CO_2$ in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.12
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• pp.1614-1619
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• 2009

The present study was conducted to observe the effect of increased osmolality of basic tris extender supplemented with trehalose and sucrose on post-thawing quality (motility, progressive motility, viability, the rate of acrosome abnormality, total abnormality and membrane integrity) of Markhoz goat spermatozoa. Fresh semen samples were evaluated for motility and sperm concentration. Only semen samples with motility more than 70% and sperm concentration higher than $3{\times}10^{9}$ sperm/ml were used for cryopreservation. In Exp. 1, trehalose (50, 75 or 100 mM) and sucrose (40, 60 or 80 mM) were added to a basic tris diluent. Based on the results of experiment 1, the goal of Exp. 2 was to investigate the combinational effects of the highest and lowest concentrations ($T_{100}+S_{80}$ or $T_{50}+S_{40}$) of trehalose and sucrose. As the control, semen was diluted and frozen in the tris diluent without trehalose or sucrose. The results in Exp. 1 showed that all evaluated spermatozoa characteristics improved significantly after freezing and thawing (p<0.05) and at the same time the increase of trehalose and sucrose concentrations in basic extenders was seen, with the best results obtained for extenders containing 70 and 100 mM trehalose and 80 mM sucrose. Comparing these results with those of control diluents, the effects of supplementation were significantly (p<0.05) better. In Exp. 2, the results showed no significant differences (p>0.05) between $T_{100}+S_{80}$ and $T_{50}+S_{40}$ extenders, but the results of $T_{50}+S_{40}$were slightly better than obtained with $T_{100}+S_{80}$ diluents. Furthermore, the results of this experiment indicated that the sperm characteristics in the isotonic control extender were significantly (p<0.05) lower than examined extenders. In conclusion, the results of this study indicated that goat sperm can tolerate hypertonic trehalose and sucrose solutions better than isotonic control diluents in the freezing period. In particular, these positive effects have been shown for acrosome integrity, which is very important for the fertilization capacity of sperm. The data indicated that addition of trehalose plus sucrose to the freezing extender can be recommended for cryopreservation of goat spermatozoa, but more data is needed on pregnancy rate, acrosome reaction and IVF to ascertain the real effect.

• Journal of the Korean GEO-environmental Society
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• v.13 no.2
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• pp.5-17
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• 2012

The amount of snow due to global warming and climate change has recently increased. The effective management of snow removal materials will be required. First, on the basis of domestic winter temperature, it is not necessary to get a baseline to less than the freezing point of $-52^{\circ}C$ for the calcium chloride($CaCl_2$) of 30%.. In terms of cost comparison between calcium chloride($CaCl_2$) and sodium chloride($NaCl$), the calcium chloride($CaCl_2$) is 2 to 3 times more expensive, and the supply of the calcium chloride($CaCl_2$) is not produced in domestic country and is in the conditions that have to imports all needed. Accordingly, the effective use scheme of snow removal materials should be considered to multifaceted ways. Thus, the objective of this study is to develop effective method and to replace from calcium choride($CaCl_2$) to sodium chloride($NaCl$) solution in the current snow removal operating system that uses a pre-wetted salt spreading method. The effective method that equals to the quality of the existing snow removal materials was developed in this study through performance tests for deicing chemicals, corrosion test of steel and freezing and thawing tests of concrete.

• Korean Journal of Animal Reproduction
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• v.11 no.3
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• pp.155-160
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• 1987

This study was carried out to determine the effects of cryoprotectants, freezing and thawing rates on the survival of 8-cell mouse embryos. The female ICR mice were induced to superovulated by intraperitoneal injections of 5 i.u. PMSG and 5 i.u. HCG given 48h apart and then were paired with males of the same strain. They were killed and embryos were flushed from the oviducts and uteri on 3 days after injection of HCG. Embryos were flushed with modified Dulbecco's phosphate buffered saline and equilibrated with 1.5M-dimethyl sulfoxide (DMSO) or 1.5M-glycerol by 3-step procedure. The freezing rates of the embryos were 1$^{\circ}C$/min from room temperature to -5$^{\circ}C$ and the embryos were seeded at -5$^{\circ}C$. After being held for 3 min at the seeding temperature, the rates were 0.3$^{\circ}C$/min from -5$^{\circ}C$ to -35$^{\circ}C$. From -35$^{\circ}C$ to -7$0^{\circ}C$, the rates were divided into 0.1$^{\circ}C$/min, 1$^{\circ}C$/min and 1$0^{\circ}C$/min, respectively. After being held for 5 min at -7$0^{\circ}C$, the embryos were plunged directly into liquid nitrogen. The embryos were thawed at 4$^{\circ}C$/min and 12$^{\circ}C$/min from -196$^{\circ}C$ to 37$^{\circ}C$, and for 2 min in 37$^{\circ}C$ water bath, respectively. The average number of ovulation points and embryos recovered were 42.7 and 34 appearing 79.5% recovery rate. Eight cell embryos in the embryos recovered were 26.3. The survival rates of embryos according to the freezing rates in the presence of 1.5M-DMSO were 73.5~80.6% at 0.1$^{\circ}C$/min, 75.0~79.5% at 1$^{\circ}C$/min and 52.8~54.7% at 1$0^{\circ}C$/min, but in the presence of 1.5M-glycerol were 62.9~67.6% at 0.1$^{\circ}C$/min, 61.4~68.3% at 1$^{\circ}C$/min and 25.5~30.2% at 1$0^{\circ}C$/min. The survival rates of embryos were not affected by the thawing rates.

• Journal of the Korea Concrete Institute
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• v.15 no.2
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• pp.197-208
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• 2003

This paper is to determine the possibility of re-using waste concrete from Incheon city area. The strength test was conducted with five aggregate compounds which was replaced a natural aggregate with recycled aggregate. After checking the physical characteristics of recycled aggregate compounds, the mix design of recycled concrete was conducted. For the relatively comparison between natural and recycled compounds, while the unit aggregate weight was changed, other conditions were fixed. The freezing and thawing test which included fly-ash and super-plastezer were performed to check the durability and workability when recycling waste concrete. In the physical characteristics of recycled aggregate, it was found that the specific gravity of recycled coarse aggregate and recycled fine aggregate satisfied the first grade of recycle specification(KS), and all compounds of recycled aggregate also satisfied the second grade of absorption specification, Especially up to the 50% substitution of recycled aggregate is equal to or a bit lower than that of convention aggregate. In comparison with conventional concrete, the recycled concrete is lower than maximum by 7% in compressive strength decreasing rate after freezing-thawing test. From now, although most of recycled concrete was used to the building lot, subgrade, asphalt admixture, through the result. It was proved that possibility of re-using recycled aggregate as the substructure of bridge, retaining wall, tunnel lining and concrete structure which is not attacked the drying shrinkage severely.

• Journal of the Korean GEO-environmental Society
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• v.21 no.9
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• pp.25-32
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• 2020

The underground utilities installed under the ground is an important civil engineering structure, such as water supply and sewerage pipes, underground power lines, various communication lines, and city gas pipes. Such underground utilities can be exposed to risk due to external factors such as concentrated rainfall and vehicle load, and it is important to select and construct an appropriate backfill material. Currently, a method mainly used is to fill the soil around the underground utilities and compact it. But it is difficult to compact the lower part of the buried pipe and the compaction efficiency decreases, reducing the stability of the underground utilities and causing various damages. In addition, there are disadvantages such as a decrease in ground strength due to disturbance of the ground, a complicated construction process, and construction costs increase because the construction period becomes longer, and civil complaints due to traffic restrictions. One way to solve this problem is to use a liquid filler. The liquid filler has advantages such as self-leveling ability, self-compaction, fluidity, artificial strength control, and low strength that can be re-excavated for maintenance. In this study, uniaxial compression strength test and fluidity test were performed to characterize the mixed soil using marine clay, stabilizer, and in-situ soil as backfill material. A freezing-thawing test was performed to understand the strength characteristics of the liquid filler by freezing, and in order to examine the effect of the filling materials on the corrosion of the underground pipe, an electrical resistivity test and a pH test were performed.

• Korean Journal of Food Science and Technology
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• v.17 no.6
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• pp.500-505
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• 1985

A method to store concentrated milk in the liquid state at $-15^{\circ}C$ was developed, and quality changes during storage of milk were evaluated. Combined cryoprotectants (CCP) suitable for storing concentrated milk in the liquid state at $-15^{\circ}C$ were consisted of 17.74% sucrose, 8.87% glucose, 8.87% fructose, 2% glycerol, 0.25% sodium hexametaphosphate, 0.25% NaCl and 0.02% ascorbic acid. The amount of CCP to be added to concentrated milk to depress freezing point to $-15^{\circ}C$ was 38% by weight. Gelation due to protein denaturation was the most serious quality change during storage, which adversely affected appearance and utilization of the stored product. Gelation was observed after 3 weeks storage in the control, but it was not in milk with CCP throughout 18 weeks storage. Amount of protein precipitated increased in the control during storage, whereas there was no protein precipitated in milk with CCP. Surface color and peroxide value of the control and treatment did not change significantly during storage, and there were no marked differences between the control and treatment. These results indicated that quality of concentrated milk could be preserved, without gelation, by storing milk with CCP in the -liquid state at the frozen storage temperature. Besides, energy required for freezing preservation of milk could be significantly reduced by elimination of phase changes for freezing and thawing, and the stored product could be continuously processed for the final products without long waiting time for thawing.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.235-240
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• 2014

Ethylene glycol (EG) has been successfully used as a cryoprotectant for vitrification of mammalian embryos (including human embryos) due to its low formula weight and high permeation into cells compared with other cryoprotectants, including propylene glycol (PROH). Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. 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). Female ICR mice (6~8 weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48 h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only during cryoprotectant step (1~4 step) but also in a variety of media (HTF, IVF medium, D-PBS) and cell stage. The results were as follows : There is no clear advantage in these freezing media of rapid method, but 4 cell and 8 cell of slow method (2, 3 and 4 step) have advantage in D-PBS. The development of embryos according to cell stage become greater in 8 cell stage. In the treatment steps of cryopreservation, the development of embryo to blastocyst was similar among rapid method, but the development of 4 cell and 8 cell embryos to blastocyst according to slow method was better than rapid method.

• Korean Journal of Heritage: History & Science
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• v.44 no.3
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• pp.78-91
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• 2011

The Usuki Stone Buddha Statues in Japan are carved on mainly dark gray welded lapilli tuff accompanied by lenticular fiamme. This rock is composed of matrix which contains feldspar and opaque minerals with some phenocrysts of quartz and feldspar. The matrix is slight to highly welded. The statues have been weathered and weakened by salt and freezing of water. To enhance the mechanical properties of the rock, consolidants and water repellents were applied. The absorption ratio of the rock was highly decreased after the treatment of the water repellents, the consolidant OH 100, as well. Ultrasonic velocity revealed similarly higher values in the treated rock by KSE 300 and OH 100, compared to non-treated rock. KSE 300, especially, highly increased the Equotip surface hardness. All studied consolidants and water repellents were found to change the original color of the stone. SNL, specifically, resulted the significant change in color. In addition, KSE 300 were observed to improve resistance to weathering such as microcrack and fracture through freezing-thawing test after treatment.