• Journal of Embryo Transfer
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• v.29 no.1
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• pp.13-19
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• 2014

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in $38.5^{\circ}C$, 5% $CO_2$ incubator. For freezing, Day 7~9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to $-7^{\circ}C$, and the straw was seeded during 8 minutes-holding time, and was cooled to $-35^{\circ}C$ at the cooling rate of $0.3^{\circ}C/min$, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method ($56.86{\pm}26.53%$) were slightly higher than those by the slow-rate freezing method ($55.07{\pm}26.43%$) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were $72.65{\pm}18.3%$ and $79.06{\pm}17.8%$, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were $66.22{\pm}18.8%$ and $45.76{\pm}12.8%$, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

• Journal of the Korean Society of Food Science and Nutrition
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• v.36 no.11
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• pp.1444-1450
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• 2007

To prevent the deterioration of jujube wine quality due to commercial heating process for sterilization, hydrostatic pressure (500 MPa, 5 min) or freezing (frozen at $-20^{\circ}C$ for 3 days, followed by thawing at room temperature for 4 hr) treatment was applied. Their microbial count, physicochemical property and sensory quality were investigated in comparison to heat-treated jujube wine ($63^{\circ}C$, 10 min) and commercial wine. Pressure-treated and commercial jujube wine were completely sterilized and heat-treated wine was decreased to <10 CFU/mL while freezing-treated jujube wine was partially sterilized $(30{\sim}60%)$. Hydrostatic pressure and freezing, and heat treatment had no influence on chemical compositions such as pH, acidity, amino acidity, reducing sugar and ethanol content, but significantly induced the changes of instrumental color. While sensory quality of heat-treated jujube wine was significantly deteriorated, reducing sour and burning taste, that of pressure and freezing-treated jujube wine was maintained fresh without decrease in sour and burning taste.

• International Journal of Highway Engineering
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• v.18 no.1
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• pp.43-55
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• 2016

PURPOSES : This study aims to develop a repair material that can enhance pavement performance, inducing rapid traffic opening through early strength development and fast setting time by utilizing MgO-based patching materials for repairing road pavements. METHODS : To consider the applicability of MgO-based patching materials for repairing domestic road pavements, first, strength development and setting time of the materials were evaluated, based on MgO to $KH_2PO_4$ ratio, water to binder ratio, and addition ratio of retarder (Borax), by which the optimal mixture ratio of the developed material was obtained. To validate the performance of the developed material as a repair material, the strength(compressive strength and bonding strength) and durability (freezing, thawing, and chloride ion penetration resistance) was checked through testing, and its applicability was evaluated. RESULTS : The results showed that when an MgO-based patching material was used, the condensation time was reduced by 80%, and the compressive strength was enhanced by approximately 300%, as compared to existing cement-based repair materials. In addition, it was observed that the strength (compressive strength and bonding strength) and durability (freezing and thawing, and chloride ion penetration resistance) showed an excellent performance that satisfied the regulations. CONCLUSIONS : The results imply that an emergent repair/restoration could be covered by a rapid-hardening cement to meet the traffic limitation (i.e. the traffic restriction is only several hours for repair treatment). Furthermore, MgO-based patching materials can improve bonding strength and durability compared to existing repair materials.

• International Journal of Highway Engineering
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• v.19 no.2
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• pp.7-15
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• 2017

PURPOSES : Infiltration of moisture, polluted material, and deicer into concrete, accompanied by freeze and thaw can cause significant deterioration of concrete pavement. In order to protect concrete from deterioration, it is necessary to prevent the infiltration of these concrete external materials. The moisture-repellent agent, which is a surface treatment and maintenance material added to concrete structures to render them water resistant, has advantages such as prevention of water infiltration and security against air permeation. Nano-coat, which is referred to as silicon hydride, is typically used as a moisture-repellent agent. Therefore, in this study, an attempt is made to use penetration-type Nano-coat as an alternative in order to evaluate its applicability through environmental resistance tests. METHODS : This study aimed to evaluate the applicability of penetration-type Nano-coat, which can provide water repellency to concrete, in concrete pavements, through various environmental resistance tests such as freezing and thawing resistance, chloride ion penetration resistance, and surface scaling resistance tests. The applicability of penetration-type Nano-coat was demonstrated based on the specification of KS F 2711, KS F 2456, and ASTM C 672. RESULTS :In the case of penetration-type Nano-coat applied on sound concrete, an increase in concrete durability was demonstrated by the negligible chloride ion penetrability and the absence of scaling, as revealed by visual observation of the surface, after 50 cycles of scaling resistance test. In addition, test result of the application of penetration-type Nano-coat on deteriorated concrete established that concrete surface pretreated by grinding provided improved durability than non-treated concrete. CONCLUSIONS :This study indicates that penetration-type Nano-coat is applicable as an effective alternative, to increase the durability of concrete structures. In addition, it was known that pretreatment of deteriorated concrete surface, such as grinding, is required to improve the long-term performance of concrete pavement.

• KSBB Journal
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• v.16 no.6
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• pp.626-631
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• 2001

Lactobacilli have been considered to play important roles in the health of human vagina. They secrete inhibitory substances to prevent vaginal infection by pathogenic organisms. In a previous study, we have isolated several lactobacilli from Korean woman and one of them (KLB46) was selected and indentified as Lactobacillu crispatus which showed high antimicrobial activity. In this study. cold adaptation prior to subsequent stresses exposure was examined whether L. crispatus KLB46 maintain the viability better than the non-adapted calls under stresses. For pharmaceutical formulation, the lyophilization process is required where stresses such as freezing/thawing and dehydration are routinely applied. Formulated L. crispatus KLB46 can be used for ecological treatment of bacterial vaginosis. The response of cold-adapted cells to other environmental stresses such as acid, heat, ethanol, NaCl, and H$_2$O$_2$ was also examined. The results showed that cold-adapted cells maintained higher survival rate compared with the non-adapted cells (freezing-thawing. 3-folds; dehydration: 3-folds; acid, 3-folds; heat, 10-folds). However, we did net observe any positive effect of cold adaptation on other stresses such as ethanol, NaCl and H$_2$O$_2$. When chloramphenicol was added during cold adaptation, adaptation effect was abolished. This confirms that de novo protein synthesis is necessary during the adaptation process. Moreover, we have identified cold shock protein homolog that codes for a major cold shock protein by PCR amplification using degenerate primers.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.98-98
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• 2003

Recently, human embryonic stem (hES) cells have become very important resources for ES cell basic research, cell replacement therapy, and other medical applications; thus, efficient cryopreservation methods for these cells are needed. This study examined whether a newly developed minimum volume cooling (MVC) vitrification method, which was tested through cryopreservation of sensitive bovine oocytes, can be used for freezing hES cells. Feeder-free cultured hES cell (MB03) colonies were mechanically dissected into several small clumps following enzymatic treatment. We compared the freezing efficiency of a slow-cooling method using a cryo-module (0.4-0.6C/min, 20-30 clumps/vial) and MVC vitrification using a modified 0.5-ml French mini-straw designated as a MVC straw (>$20,000{\circ}C$/min, 10 clumps/straw) After thawing, in vitro survival of hES cell clumps was higher for MVC-vitrified cells (80.8%, 97/120) than for slow-cooled cells (38.2%, 39/102). Further, the proliferation rate of surviving MVC-vitrified cells was similar to that of control hES cells from 2 weeks after thawing. In addition, vitrified-thawed hES cells demonstrated a normal karyotype, were positively immunostained for surface marker antibodies (AP, SSEA-4 and TRA-1-60) and the Oct-4 antibody, and could differentiate into all three embryonic germ layer cells in vitro. This result demonstrates that hES cell clumps can be successfully cryopreserved by a newly developed MVC vitrification method without loss of human cell characteristics.

• 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.

• Korean Journal of Animal Reproduction
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• v.23 no.2
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• pp.165-174
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• 1999

Boar semen can be frozen successfully. However, there is a large variability in the extent of damage boar semen samples experiences during cryopreservation. This experiment was undertaken to find out factors that affect a post-thaw viability of boar spermatozoa. For this purpose, cryodiluents(BF5, LEY, Soejima and M-Soejima), cryoprotectants(glycerol. ethylene glycol, and propylene glycol), pre-freezing method(dryice-pellet, dryice-straw and L$N_2$vapour-st-raw) and total time required for freezing(2. 5, and 7 h) were compared as a factors. To investigate quality of semen during freezing process, motility(%), normal apical ridges(%, NAR), and proportion of living sperm(%) by flow cytometic analysis were assessed after collection, cooled, pre-frozen and post-thawing. Post-thaw motility of semen diluted with M-Soejima was 52.0%, respectively. When heparin, caffeine or heparin＋caffeine was added to 2nd cryodiluent of M-Soejima during freezing process, the highest motility after thawing was shown at the addition of caffeine (2mM), with 61.7$\pm$2.9% of motility. M-Soejima with heparin or caffeine was significantly higher than that of controI(p＜0.05). The result using glycerol(Gly), ethylene glycol(EG), propylene glycol(PG), and their mixture (Gly＋EG and Gly＋PG) as cryoprotectants, the highest motility was shown at the mixture treatment with Gly plus PG. However, the highest proportion of live spermatozoa was shown at Gly＋EG, there was no significantly difference among treatments(p＞0.05). When semen was pre-frozen with three manners(dryice-pellet, dryice-straw, and L$N_2$ vapor-straw), motility(%) of post-thaw spermatozoa was the highest in the L$N_2$ vapor-straw pre-freezing method of M-Soejima cryodiluent with 57.5% of motility, For a simple, economical and timesaving approach to freezing boar semen, total time required for freezing were 2, 5, and 7 hours, post-thaw motility were 43.8, 45.0 and 38.8%, NAR were 19.5, 22.7 and 28.5%, and viability were 20.8, 19.9 and 22.1%, respectively. This data suggests that boar semen diluted with M-Soejima cryodiluent contained caffeine, using mixture of glycerol and propylene glycol or ethylene glycol as cryoprotectants, frozen with 2 hours, can be taken better motility, NAR, and proportion of live spermatozoa.

• Journal of Food Hygiene and Safety
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• v.10 no.1
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• pp.23-28
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• 1995

The effect of the ethanol extract from salviae miltiorrhizae radix (Salvia miltiorrhiza) on the microbial growht and the stability of the extracted antimicrobial material were investigated. The ethanol extract had strong growth inhibition activity (MIC, 3.13~50.0 $\mu\textrm{g}$/ml) against Gram-positive bacteria such as B. subtilis, L. monocytogenes and S. aureus. Among Gram-positive bacteria tested, B. subtilis was the most susceptible to the extracted substance. While the antimicrobial activity of the ethanol extract was weak (MIC, 400~800 $\mu\textrm{g}$/ml) to E. coli and yeasts (C. albicans, Sacch. diastaticus). The ethanol extract had bactericidal action at higher concentration than MIC against B. subtilis, while the extract had only bacteriostatic action against S. aureus. The extracted antimicrobial substance was stable in the pH range of 4.0 to 10.0, heat treatment at 121$^{\circ}C$ for 15 min, and freezing and thawing

• Clinical and Experimental Reproductive Medicine
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• v.26 no.2
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• pp.127-135
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• 1999

Cryopreservation is able to store the surplus pre-embryos for freezing and furthermore thawing and transfer in a subsequent cycle. Cryopreserving cells which are maintaining their viability are the very complex process. This study has been carried out in order to find the effects of cryopreservation steps, freezing media and embryonic stages on the rates of viability and development of cryopreserved mouse embryos. Female ICR mice ($6{\sim}8$ weeks old) were induced to superovulate by sequential intraperitoneal injection of 5 IU PMSG and 5 IU hCG 48h apart. Mouse embryos were collected according to its developmental stage after the injection of hCG. Embryos were cryopreserved not only by cryoprotectant step (1 step${\sim}$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, 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.