• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.78-78
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• 2002

The role of heat shock proteins in shielding organism from environmental stress is illustrated by the large-scale synthesis of these protein by the organism studied to date. However, recent evidence also suggests an important role for heat shock protein in fertilization and early development of mammalian embryos. Effects of elevated in vitro temperature on in vitro produced bovine embryos were analysed in order to determine its impact on the expression of heat shock protein 70 (HSP70) by control and frozen-thawed after in vitro fertilization (IVF) or nuclear transfer (NT). The objective of this study was to assess the developmental potential in vitro produced embryos with using of the various containers and examined expression and localization of heat shock protein 70 after it's frozen -thawed. For the vitrification, in vitro produced embryos at 2 cell, 8 cell and blastocysts stage after IVF and NT were exposed the ethylene glycol 5.5 M freezing solution (EG 5.5) for 30 sec, loaded on each containers such EM grid, straw and cryo-loop and then immediately plunged into liquid nitrogen. Thawed embryos were serially diluted in sucrose solution, each for 1 min, and cultured in CRI-aa medium. Survival rates of the vitrification production were assessed by re-expanded, hatched blastocysts. There were no differences in the survival rates of IVF using EM grid, cryo-loop. However, survival rates by straw were relatively lower than other containers. Only, nuclear transferred embryos survived by using cryo-loop. After IVF or NT, in vitro matured bovine embryos 2 cell, 8 cell and blastocysts subjected to control and thawed conditions were analysed by semiquantitive reverse transcription polymerase chain reaction methods for hsp 70 mRNA expression. Results revealed the expression of hsp 70 mRNA were higher thawed embryos than control embryos. Immunocytochemistry used to localization the hsp70 protein in embryos. Two, 8-cell embryos derived under control condition was evenly distributed in the cytoplasm but appeared as aggregates in some embryos exposed frozen-thawed. However, under control condition, blastocysts displayed aggregate signal while Hsp70 in frozen-thawed blastocysts appeared to be more uniform in distribution.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.313-321
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• 1999

The purpose of these study was to investigate the use of a glass micropipette (GMP) as a vessel for vitrification of bovine IVP blastocysts, to compare the post-thaw survival rates of bovine blastocysts frozen in GMP with those frozen in OPS that have been previously investigated, and to improve the hatching rate following vitrification with GMP method. The GMP vessel permits higher freezing and warming rate than the OPS due to the higher heat conductivity of the glass and lower mass of the solution that contains the embryos. Groups of three bovine IVP blastocysts were sequentially placed into vitrification solution before being loaded into either the OPS or GMP vessels and immersed into L$N_2$within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for each 5 min, respectively, and then cultured in TCM 199 supplemented with 10% FCS for 24 h. The rate of blastocyst re-expanding did not significantly different for OPS (75.9%) and GMP (80.0%) methods (P＞0.05). The hatching rates in OPS (34.1%) and GMP (37.5%) methods were significantly lower than that in control group (54.3%) (P＞0.05). In addition, the rate of blastocyst re-expanding was significantly lower if blastocysts were vitrified in the wide portion of the micropipette rather than the narrow portion of the micropipette (83.3 vs 56.7%) (P＞0.05), even though three blastocysts were loaded per vessel. The hatching rate in 0.05% pronase solution treatment for 30, 60 and 90 see (45.9, 54.7 and 57.5%) were significantly higher than that in control (35.0%), even though there was not significantly different between 30 see and control. These results indicate that both vitrification vessels can provide high survival rates of bovine IVP blastocysts. However, the GMP vessel has the advantage over the OPS, in that the former does not need a cap to protect the vessel from floating after immersion in L$N_2$. The location of the embryos (narrow or wide portion of immersion) were considered to be limiting factors to the viability of bovine IVP embryos. The exposing in 0.05% pronase solution for 60 or 90 see can increase hatching rates of post-thaw bovine IVP blastocysts.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.21 no.6
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• pp.527-535
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• 2020

The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell Keep^TM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell Keep^TM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell Keep^TM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell Keep^TM was determined to be 50% and can be considered as a proper preservation method for cryobanking.

• Cartoon and Animation Studies
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• s.37
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• pp.197-219
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• 2014

, one of the Disney's animations, hit the 10 million audience mark for the first time in the history of animations released in Korea. not only raised the fever with its theme song, 'Let it go', as well as Elsa, Anna, and Olaf's character products but caused sensations in many ways. If so, we need to think about what kind of meaning did create in Korea to be so sensational. This study examines the value that Frozen intended to deliver and the meaning it generated by using Greimas actant model and semiotic square. From the actant model analysis on Anna and Elsa from , it was identified that Anna desired to recover her relationship with Elsa and to take summer back in Arendelle. Her desires can be interpreted as her love toward Elsa and people in Arendelle. Meanwhile, Elsa always desired freedom although she confined herself because of her ability to freeze. In other words, Elsa desired to free herself from her freezing ability by finding out how to control her ability. Such desires of Anna and Elsa were achieved by their actions of true love, and the solution of all the conflicts in was an action of true love. From the semiotic square analysis on the meaning of , it was found out that created past-oriented value with which characters tried to change their abnormal lives of the present into their normal lives of the past. The characters tried to change their present lives where freezing winter comes in the middle of summer, communication between the sisters is cut off, and people try to take advantage of the abnormal state deliberately, into the past when the sisters had a good relationship and the natural season of summer in Arendelle. The past-oriented value that tried to tell us is similar to our reality. In our reality with a lot of unbelievable news and unstable circumstances, we desire to go back to the past when we were filled with affection and hope even though our lives were tough and difficult. This sentiment must have contributed to the huge success of in Korea.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.10 no.4
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• pp.189-197
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• 1977

In present study, the effects of various factors including the solvent concentration, extraction time and temperature, the ratio of sample vs extraction solvent, (w/v) and pH upon the extractability of the NaCl and alcohol soluble proteins of marine algae were investigated. Eight species of fresh algae, the major ones in consumption as food, namely Porphyra suborbiculata, Undarie pinnatifida, Hizikia fusiforme, Sargassum, fulvellum, Enteromorpha linza, Sargassum kjellmanianum, Codium coarctatum, and Ulva pertusa were used for the extraction of NaCl soluble protein and dried materials of four species, Perphyra suborbiculata, Undaria pinnatifida, Enteromorpha linza and Sargassum fulvellum were used for the extraction of alcohol soluble protein. The frozen and mascerated samples were prepared by the same method described in previous paper (Ryu, 1977). And the dried materials were moistened with alcohol solution before freezing. The effect of solvent concentration on the extractability of NaCl soluble protein differed from species. The extractability of Undaria Pinnatifide, Hizikia fusiforme, Perphyra suborbiculata, Enteromorpha linza, and Ulva pertusa reached maxima at 0.25M NaCl solution while the 1.0M for Sargassum fulvellum, Saygassum kjellmanianum and Codium coarctatum. In case of alcohol soluble proteins, it was shown at $20\%$ ethanol solution for Porphyra suborbiculata, Undaria pinnatifida, Enteromorpha linza, and Sargassum fulvellum. Variation of the ratio of sample vs solvent gave slight effect upon the extractability, but the ratio of 1:30(w/v) seemed most efficient for the extraction of NaCl soluble proteins and 100 ml solvent added to 1 g dried sample was effective in case of alcohol soluble proteins. Extraction time has a minimal effect upon the extraction of alcohol soluble protein, and approximately 21 to $43\%$ of algal protein was extracted within 1 hour. But in case of NaCl soluble protein extraction, the effect of time revealed differently from species to species resulting in that the extraction for 1 hour gave a maximum extractability in Ulva pertusa and Enteromorpha linza, 2 hours in Porphyra suborbiculata, Codium coarctatum and 3 hours in Undaria pinnatifica, Hizikia susiforme, Sargassum fulvellum and Sargassum kjellmanianum. When the NaCl soluble protein of Undaria pinnatifida and Enteromopha linza was extracted at various temperature, the most effective extraction temperature was $40^{\circ}C$ while the temperature was $50^{\circ}C$ for Undaria pinnatifida and $60^{\circ}C$ for Hixikia fusiforme, Sargassum fulvellum, Sargassum kjellmanianum and Codium coarctatum. Bus in case of alcohol soluble extraction, the optimum temperature was $30^{\circ}C$ for Enteromorpha linza and $40^{\circ}C$ for Undaria pinnatifida, Sargassum fulvellum and Porphyra suborbiculata. In the effect of pH on extractability, the maximum extractability of NaCl soluble proteins was obtained at pH 7to 8 and pH 8 to 9 for alcohol soluble protein.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.352-362
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• 1985

In succession to the previous paper, the present study was directed to investigate the optimal processing conditions of meat paste products with mackerel. To improve the gel forming ability of meat paste, washing conditions with water and alkaline solution, setting time and temperature, and heating temperature were controlled, and the influences of the freshness of raw mackerel and mixing ratios of ordinary and dark muscle on the qualities of meat paste products were discussed. The most effective condition for the keeping freshness of raw mackerel meat among different storage conditions was the forozen storage at $-20^{\circ}C$, followed by the storage at $-3^{\circ}C$ and ice storages, and this relation was coincided with the effect for maintaining of gel forming ability among above conditions, but there was no effect on keeping freshness of raw mackerel in the storage at $25^{\circ}C$. Gel strength of meat washed with tap water decreased with washing time, particularly, the meat washed three times showed higher gel strength than the meat washed more than 5 times. And the removal ratios of water soluble protein were $60\%$ in the meat washed three times and $90\%$ in the meat washed nine times. Washing effect of raw mackerel meat with alkaline solution was great at pH $6.5{\sim}7.0$ of meat paste yielding the highest gel strength in the meat washed with $0.5\%$ sodium bicarbonate solution. Gel strength of meat paste product decreased with the increase of mixing ratios of ordinary and dark muscle in the raw meat. In the setting conditions of meat paste examined, 15 hours at $5^{\circ}C$ and 2 hours at $30^{\circ}C$. The most suitable temperature for gel forming in heating conditions was $90^{\circ}C$, fellowed by $100^{\circ}C\;and\;80^{\circ}C$.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.18 no.4
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• pp.339-351
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• 1985

This study was carried out to investigate the optimal conditions for meat paste production with sardine. To improve the gel forming ability of meat paste, washing time and condition with alkaline solution, setting time and temperature, and heating temperature before pasteurization were controlled, and the influences of the freshness of raw sardine and the mixing ratios of ordinary and dark muscles on the duality of the meat paste product were discussed. The frozen storage showed a predominant effect on keeping freshness of raw sardine at different storage conditions and gel forming ability was maintained for 1 day at ice storage, for 3 days at $-3^{\circ}C$ and for 4 days at frozen condition, but there was no effect on keeping freshness of raw sardine in the storage at $25^{\circ}C$. Gel strength of meat paste product tended to decrease with washing time of raw meat, and in case of washing 3 times the meat appeared excellent in gel strength, but in case of seven and nine times the meat showed lower water holding capacity and decreased organoleptic test score in the quality of meat paste prtoduct. Raw meat washed with alkaline solution showed a desirable effect on gel forming ability compared with that washed with tap water, and in the case of washed with $0.5\%$ sodium bicarbonate solution exhibited the most favorable effect on gel forming. The gel strength of the meat paste product decreased with the increase of mixing ratios of dark muscle in the raw meat. Setting time and temperature for the gel forming ability of meat paste were good at $5^{\circ}C$ for 20 hours and at $20^{\circ}C$ for 2 hours. In the heating temperature of meat paste, heating treatment at $90^{\circ}C$ was desirable for gel forming.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.85-92
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• 1999

This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.

• Development and Reproduction
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• v.20 no.3
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• pp.219-225
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• 2016

Most of the commercial devices for vitrification are directly immersed into the warming solution (WS) for increasing of warming rate. However, the previous modified cut standard straw (MCS) which has reported is difficult to immerse into the WS. The aim of this study was to investigate whether the long cut straw (LCS) could be useful as a stable tool for vitrified-warmed human blastocysts. A total of 138 vitrified-warmed cycles were performed between November 2013 and November 2014 (exclusion criteria: women ${\geq}38$ years old, poor responder, surgical retrieval sperm, and severe male factor). The artificial shrinkage was conducted using 29-gauge needles. Ethylene glycol and dimethyl sulfoxide (7.5% and 15% (v/v)) were used as cryoprotectants. Freezing and warming were conducted using the LCS tool. The cap of LCS was removed using the forceps in the liquid nitrogen ($LN_2$) and then directly immersed into the first WS for 1 min at $37^{\circ}C$ (1 M sucrose). Only re-expanded blastocysts were transferred after it was cultured in sequential media for 18-20 h. A total of 294 blastocysts were warmed, and all were recovered (100%). Two hundred eighty-five embryos were survived (96.9%). The vitrified-warmed blastocysts of all patients were transferred without any cancellation. We were able to achieve a reasonable implantation (24.2%), following by clinical pregnancy (36.2%), which then continued to ongoing pregnancy (36.2%), and live birth (31.2%). Using LCS is achieved the acceptable rates of survival, pregnancy and live birth. Therefore, the LCS could be considered as a stable and simple tool for human embryo vitrificaton.

• Journal of the Korea Concrete Institute
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• v.18 no.1 s.91
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• pp.57-64
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• 2006

The crack presented in concrete structures causes a structural defect, the durability decrease, and external damages etc. Therefore, it is necessary to improve durability through the effort to control the crack. Fluosilicic acid($H_2SiF_6$) is recovered as aqueous solution which absorbs $SiF_4$ produced from the manufacturing of industrial-graded $H_3PO_4$ or HF. Generally, fluosilicates prepared by the reaction between $H_2SiF_6$ and metal salts. Addition of fluosilicates to cement endows odd properties through unique chemical reaction with the fresh and hardened cement. Mix proportions for experiment were modulated at 0.45 of water to cement ratio and $0.0{\sim}2.0%$ of adding ratio of fluosilicate salt based inorganic compound. To evaluate correlation of concrete strength and adding ratio of fluosilicate salt based inorganic compound, the tests were performed about design strength(21, 24, 27 MPa) with 0.5% of adding ratio of fluosilicate salt based inorganic compound. Applications of fluosilicate salt based inorganic compound to reduce cracks resulted from plastic and drying shrinkage, to improve durability are presented in this paper. Durability was evaluated as neutralization, chloride ion penetration depth, freezing thawing resistant tests and weight loss according reinforcement corrosion. It is ascertained that the concrete added fluosilicate salt based inorganic compound showed m ability to reduce the total area and maximum crack width significantly as compared non-added concrete. In addition, the durability of concrete improved because of resistance to crack and watertightness by packing role of fluosilicate salt based inorganic compound obtained and pozzolanic reaction of soluble $SiO_2$ than non-added concrete.