• Clinical and Experimental Reproductive Medicine
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• v.27 no.4
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• pp.367-372
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• 2000

Objective: This study was performed to evaluate whether vitrification method could be used for the cryopreservation of human blastocysts derived from IVF program. Methods: Surplus embryos were obtained from consented IVF patients. Controlled ovarian hyperstimulation was done with midluteal GnRH agonist, gonadotropin and hCG. After oocyte retrieval and insemination, fresh embryo transfer was done at $4{\sim}8$ cell stage. The surplus embryos after ET were cultured in blastocyst medium up to 6 days after oocyte retrieval. Obtained blastocysts were cryopreserved with our vitrification method. Blastocysts were exposed to 1.5 Methylene glycol (EG) in phosphate buffered saline (PBS) for 2.5 minutes, followed by 5.5 M EG plus 1 M sucrose for 20 seconds. Then 1 to 3 blastocysts were mounted on electron microscope (EM) grid and the grid was plunged into liquid nitrogen for storage. For thawing, blastocyst-containing EM grids were sequentially transferred in 1.0 M, 0.5 M, 0.25 M, 0.125 M and 0 M sucrose solution at the intervals of2.5 minutes. And blastocysts were cultured for about 6 hours and only re-expanded blastocysts were transferred to uterus of the patients on 4 to 5 days after ovulation in natural cycle or on 18 to 19 day of artificial cycle. Results: From Oct. 1998 to Jul. 1999, 34 patients were agreed to participate in this study. The mean age and duration of infertility of the patients were 31.6 years and 4.1 years, respectively. Among 34 cycles. replacements could be done in 20 cycles (58.8%). A total 93 blastocysts were thawed and 48 (51.6%) of them survived. Thirty-eight blastocysts, mean 1.9 embryos per patient, were transferred, resulting in 5 clinical pregnancies which consisted of 1 triplet, 2 sets of twins and 2 singleton pregnancies. The pregnancy rate per transfer was 25% and implantation rate was 23.6%. Five patients delivered 7 healthy babies including 2 sets of twins at term. Conclusion: Successful pregnancies and deliveries were established after transfer of vitrified human blastocysts. Vitrification using ethylene glycol as cryoprotectant and electron microscope grid is a rapid and simple method that can be effectively applied for the cryopreservation of human blastocysts.

• International Journal of Highway Engineering
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• v.18 no.6
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• pp.123-130
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• 2016

OBJECTIVES : The objective of this research is to determine the moisture resistance of the freeze-thaw process occurring in low-noise porous pavement using either hydrated-lime or anti-freezing agent. Various additives were applied to low-noise porous asphalt, which is actively paved in South Korea, to overcome its disadvantages. Moreover, the optimum contents of hydrated-lime and anti-freezing agent and behavior properties of low-noise porous asphalt layer are determined using dynamic moduli via the freeze-thaw test. METHODS : The low-noise porous asphalt mixtures were made using gyratory compacters to investigate its properties with either hydrated-lime or anti-freezing agent. To determine the dynamic moduli of each mixture, impact resonance test was conducted. The applied standard for the freeze-thaw test of asphalt mixture is ASTM D 6857. The freeze-thaw and impact resonance tests were performed twice at each stage. The behavior properties were defined using finite element method, which was performed using the dynamic modulus data obtained from the freeze-thaw test and resonance frequencies obtained from non-destructive impact test. RESULTS : The results show that the coherence and strength of the low-noise porous asphalt mixture decreased continuously with the increase in the temperature of the mixture. The dynamic modulus of the normal low-noise porous asphalt mixture dramatically decreased after one cycle of freezing and thawing stages, which is more than that of other mixtures containing additives. The damage rate was higher when the freeze-thaw test was repeated. CONCLUSIONS : From the root mean squared error (RMSE) and mean percentage error (MPE) analyses, the addition rates of 1.5% hydrated-lime and 0.5% anti-freezing agent resulted in the strongest mixture having the highest moisture resistance compared to other specimens with each additive in 1 cycle freeze-thaw test. Moreover, the freeze-thaw resistance significantly improved when a hydrated-lime content of 0.5% was applied for the two cycles of the freeze-thaw test. Hence, the optimum contents of both hydrated-lime and anti-freezing agent are 0.5%.

• Journal of Animal Science and Technology
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• v.55 no.5
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• pp.427-434
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• 2013

This study was conducted to establish methods for preserving chicken primordial germ cells (PGCs) for long-term storage in liquid nitrogen and for developmental engineering or preservation of species. The purpose of this study is to clarify the effects of fetal bovine serum (FBS) or chicken serum (CS) treatment on the viability of cryopreserved PGCs from Korean Native Chicken (Ogye). PGCs separated from a germinal gonad of an early embryo at day 5.5-6 (stage 28) were suspended in a freezing medium containing freezing and protective agents (dimethyl sulfoxide (DMSO), ethylene glycol (EG) and glycerol). The values from 0, 5, 10, and 15 % DMSO plus FBS treatment were 21.6, 30.36, 36.42, 50.39, and 48.36 %, respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% EG + FCS treatment (p<0.05) (64.36% vs. 50.66%). This study establishes a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGC in liquid nitrogen at a germplasm repository and an ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted to improve the production of germline chimeras.

• Journal of Animal Science and Technology
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• v.55 no.5
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• pp.417-425
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• 2013

We sought to provide a method for freezing and preserving primordial germ cells, or an avian germ cell of a bird, as a material for developmental engineering or species preservation. The aim of this study was to compare the efficacy of slow freezing with a vitrification method for the cryopreservation of chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonad of day 5.5-6 day (stage 28) cultured chick embryos, using the MACS method, were classified into two groups: slow freezing and vitrification. We examined the viability of PGCs after Cryopreservation. Four freezing methods were compared with each other, including the following: Method 1: The PGCs were frozen by a programmed freezer in a plastic straw, including 2.0 M ethylene glycol (EG) as cryoprotective additive (slow freezing) Method 2: The PGCs were vitrified in a plastic straw, including 8.0 M EG, plus 7% polyvinylpyrrolidone (PVP) (rapid freezing). Method 3: The slow freezing was induced with a cryotube including 2.0 M EG Method 4: The PGCs were frozen in a cryotube including 10% dimethyl suloxide (DMSO) (rapid freezing). After freezing and thawing, survival rates of the frozen-thawed PGCs from Method 1 to 4were 76.4%, 70.6%, 80.5% and 78.1% (p<0.05), respectively. The slow freezing ($-80^{\circ}C$ programmed freezer) method may provide better survival rates of frozen-thawed PGCs than the vitrification method for the cryopreservation of PGCs. Therefore, these systems may contribute to the cryopreservation of a rare avian species.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.48 no.4
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• pp.411-416
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• 2015

Olive flounder Paralichthys olivaceus production has increased gradually in recent years, but prices have fallen. Thus, the development of a variety of processed foods incorporating olive flounder would help to increase the income of fishermen. This study was conducted to investigate the best method for olive flounder ball processing. Clean olive flounder were divided into five portions. Olive flounder meat (100 g with added egg white 39 g) was chopped and then mixed with 10 mL fresh cream and ingredients. The dough was molded into the shape of a ball. The olive flounder balls were then processed by two different methods. In the first method, the flounder ball was boiled in water for 3 min then vacuum-packed in polyethylene film and stored at $-20^{\circ}C$ for 7 days. After 7 days, the ball was thawed and heated in a microwave for 2 min (Sample-1). In the second method, the ball was vacuum-packed in polyethylene film without boiling and then stored at $-20^{\circ}C$ for 7 days before thawing and boiling in water for 3 min (Sample-2). After heating, both types of olive flounder balls were evaluated. Various factors (including the viable bacterial count, chemical composition, pH, hardness, thiobarbituric acid level, salinity, and free amino acid content) were measured, and a sensory evaluation was conducted. Based on the results of the sensory and hardness evaluations, Sample-1 was deemed to be superior to Sample-2.

• Clinical and Experimental Reproductive Medicine
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• v.39 no.1
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• pp.33-39
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• 2012

Objective: We devised a novel strategy, a GnRH antagonist protocol with a GnRH agonist trigger followed by frozen-thawed blastocyst transfers with long zona dissection (LZD). The purpose of this study was to investigate the clinical outcomes of this new strategy according to age. Methods: Ninety women aged less than 35 (group A) and 32 women aged 35 to 39 (group B) underwent the GnRH antagonist protocol with a GnRH agonist trigger in order to obtain many oocytes and prevent early-onset ovarian hyperstimulation syndrome (OHSS). All oocytes were cultured to the blastocyst stage and all blastocysts grade 3BB or better were cryopreserved. Embryo transfers were only performed in freeze-thaw cycles to prevent late-onset OHSS and to overcome embryo-endometrium dyssynchrony. LZD was performed just after thawing to improve hatching and implantation rates. Results: The average numbers of retrieved oocytes and blastocysts grade 3BB or better were $12.8{\pm}5.5$ and $4.4{\pm}2.6$ in group A and $10.9{\pm}7.4$ and $2.5{\pm}2.2$ in group B, respectively, and OHSS did not occur in any of the women. Implantation rates were 46.7% in group A and 39.3% in group B. Cumulative clinical pregnancy rates per retrieval were 77.8% in group A and 62.5% in group B. Cumulative ongoing pregnancy rates per retrieval were 71.1% in group A and 53.1% in group B. Conclusion: GnRH antagonist protocol with GnRH agonist trigger followed by frozen-thawed blastocyst transfers with LZD can generate many blastocysts without OHSS and maximize cumulative pregnancy rates per retrieval. This strategy is more effective in young women aged less than 35 than in women aged 35 to 39.

• Journal of Nutrition and Health
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• v.49 no.1
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• pp.43-50
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• 2016

Purpose: This study was conducted to examine prerequisite-program performance of the hospital foodservice operation and develop measures for improvement of prerequisite-program performance. Methods: Data were collected through surveys administered to 168 hospital dieticians in the Seoul, Incheon, Gyeonggi-do, Daegu, and North Gyeongsang areas. Out of total questionnaires, 65 questionnaires were usable and the response rate was 38.7%. Statistical analyses were performed using the SPSS program (ver 20.0) for ${\chi}^2-test$ and one-way ANOVA. Results: According to the result of the prerequisite-program evaluation, the percentage of 'appropriate', 'needs to be improved', and 'inappropriate' was 44.6%, 47.7%, and 7.7%, respectively. The score for the 'inappropriate' group was significantly lower than that of the 'appropriate' group or 'needs to be improved' group on the food safety management (p < 0.001), preparation facility management (p < 0.001), water management (p < 0.001), and storage transportation management (p < 0.05) parts. Holding rate of foodservice facility and equipments in the 'appropriate' group were significantly higher than in the others on convection oven (p < 0.01), air conditioner (p < 0.01), three-compartment sink (p < 0.01), hot-holding equipment (p < 0.01), cold-holding equipment (p < 0.05), exclusive thawing refrigerator (p < 0.05), and sterilizer for sanitary shoes (p < 0.05) items. Conclusion: To improve the quality of hospital foodservice, foodservice managers and HACCP specialists should develop and implement a prerequisite-program and a HACCP plan considering the characteristics of the hospital foodservice operation.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.3
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• pp.115-119
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• 2014

Objective: This study was designed to investigate the survival rate of vitrified mouse blastocysts depending on the presence or absence of sucrose in vitrification solution. Methods: Mouse two-cell embryos were collected and cultured to blastocysts. Two vitrification solutions were prepared. The control solution was composed of 25% glycerol, 25% ethylene glycol, and 0.5 M sucrose (G25E250.5S) containing 2.5 mL glycerol, 2.5 mL ethylene glycol, 2 mL SSS, and 0.855 g sucrose in 5 mL PB1. The experimental solution was composed of 25% glycerol and 25% ethylene glycol (G25E25) and contained 2.5 mL glycerol and 2.5 mL ethylene glycol in 5 mL PB1. Artificial shrinkage was conducted by aspirating the blastocoelic fluid using an ICSI pipette. To examine the effect of sucrose in the vitrification solution on the survival rate of mouse blastocysts, the shrunken-equilibrated blastocysts were rehydrated or vitrified after being exposed to one of the two vitrification solutions. After exposure and the vitrification-thawing process, the re-expansion rate and hatching rate were evaluated after 6 hours of in vitro culture. Results: The re-expansion rate of mouse blastocysts exposed to vitrification solution with and without sucrose were not different in the experimental solution (without sucrose) (98%) and the control solution (with sucrose) (92%) (p>0.05). The hatching rate was higher in the experimental solution (95%) than in the control solution (88%), but did not differ across two treatments (p>0.05). The re-expansion rate of mouse blastocysts vitrified in the control solution was 92% and 94%, respectively (p>0.05), and the hatching rate was higher in the experimental solution (90%) than in the control solution (74%) (p<0.05). Conclusion: Sucrose need not be added in vitrification solution for freezing of artificially shrunken mouse blastocysts.

• Clinical and Experimental Reproductive Medicine
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• v.46 no.4
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• pp.166-172
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• 2019

Objective: In vitro maturation (IVM) of immature oocytes can be useful for some infertile patients. In IVM programs, the rates of embryo formation and pregnancy are low. Therefore, it is essential to recognize the main factors involved in regulating oocyte maturation in vitro. The purpose of this study was to investigate the effects of growth differentiation factor 9 (GDF9) and cumulus cell (CC) supplementation in IVM medium on the rates of embryo formation and viability of human blastocysts. Methods: A total of 80 germinal vesicle oocytes from stimulated cycles underwent an IVM program. The oocytes were divided into four groups, where group I consisted of IVM media only and served as the control, group II consisted of IVM+CCs, group III consisted of IVM+GDF9 (200 ng/mL), and group IV consisted of IVM+CCs+GDF9 (200 ng/mL). Intracytoplasmic sperm injection was performed on the IVM oocytes, and the cleavage embryos that were generated were vitrified. Following thawing, the embryos were cultured for 3 additional days, and the viability rates of the developed blastocysts were determined. Results: The maturation rate of the oocytes did not differ significantly across the four groups. The fertilization rate in group II was significantly higher than that in the control group (76.5% vs. 46.2%). Embryo formation was significantly more frequent in all experimental groups than in the control group, while blastocyst formation did not show significant differences in the three experimental groups compared to the control. The mean viability rates in groups II, III, and IV were 58.16%, 55.91%, and 55.95%, respectively, versus 37.78% in the control group (p< 0.05). Conclusion: Supplementation of IVM culture media with GDF9 and CCs enhanced the fertilization, embryo formation, and viability rates of blastocysts generated from vitrified cleavage embryos.

• Journal of the Korea Concrete Institute
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• v.15 no.4
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• pp.529-540
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• 2003

Recently, the researches on the durability design of concrete structures have been studied. As the examples, models to evaluate the service life prediction of the structure have been developed. The purpose of this article is to develop the model for predicting remaining service life. The final aim is to provide the user time for repairing the concrete structures. In addition, it makes possible to maintain the concrete structure economically. 70 reservoirs out of the inland concrete structures were selected and concrete structures of their components were surveyed. Two methods were used for measuring carbonation; TG/DTA method and Phenolphtalein indicator and, the value of pH was measured by the pH meter, After deriving correlations of calcium carbonate and used year, duration from completion year to 2002, pH value, and concrete cover depth the model was developed for predicting remaining service life by measuring data as small as possible. The conventional models had been developed on the basis of experiment data obtained from the restricted lab environment like as carbon gas exposure. On the other hand this model was developed on the basis of measuring data obtained from the real field that the complex deterioration actions are occurred such as freezing and thawing, carbonation, steel corrosion, and so on. The reliability of the developed model will be evaluated high in this point and this model can help to maintain concrete structures economically by providing the manager time to repair the deteriorated concrete structures in site of facility management.