• Clinical and Experimental Reproductive Medicine
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• 제27권4호
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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.

• 한국도로학회논문집
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• 제18권6호
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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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• 제55권5호
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• pp.427-434
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• 2013

귀중한 한국재래닭 (오계)의 원시생식세포를 냉동보존하고, 키메라 닭을 통한 재래종의 복원을 도모하는 방법을 실용화하기 위해서, 오계의 원시생식세포에 있어 최적의 동결방법에 대해 검토했다. 원시생식세포의 동결은, 세포를 분리한 후, 동결배지의 기초가 되는 혈청으로서 소 태아혈청 (FBS) 그리고 닭 혈청(CS)를 이용하고, 동결보호제로서는 EG 및 DMSO의 첨가량을 2.5, 5, 10, 15%로 설정하고, 300 ${\mu}L$의 동결배지 용량으로 동결 튜브 안에서 $-70^{\circ}C$로 동결했다. 융해 후에는 오계 PGC의 생존율을 확인 및 검토를 했다. 그 결과, FBS 처리군이 CS 처리군 보다 생존 PGC 회수율이 높은 경향을 나타냈다. 또한 항동해제로서 최적의 조건은 10% EG + FBS의 조합을 기초로 한 동결배지에서 가장 높은 오계원시 생식세포의 생존율을 확인했다. 이러한 결과들로부터 한국재래종(오계)의 원시생식세포의 동결보존의 실용화가 보다 더 향상 될 수 있는 또 하나의 방법이 될 수 있음을 시사한다.

• Journal of Animal Science and Technology
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• 제55권5호
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• pp.417-425
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• 2013

동결 닭 PGCs의 생식계열 키메라를 이용한 생체에의 복원을 실용화 하기 위해서는, 닭 PGCs의 동결보존기술의 향상에 의해 동결 및 융해 후의 많은 생존세포를 확보 하는 것과, 생식계열 키메라의 제작효율을 높이는 것이 반드시 필요하다. 닭 PGCs는 배양 5.5일령의 닭 원시생식선으로부터 채취하고, ACS 방법에 의해서 순수 닭 PGCs를 분리했다. 닭 PGCs의 동결보존실험결과 다음의 4종류의 동결방법을 각각 비교 검토했다. 1. 플라스틱 스트로에 의한 완만동결법 (SF), 동결보호물질은 2M 에틸렌 글리콜 (EG), 2. 스트로에 의한 급속동결법 (RF), 8M EG + 7% PVP, 3. 동결용 Cryotube에 의한 SF, 2M EG, 4. 튜브에 의한 SF, 10% DMSO. 동결 및 융해 후의 PGCs의 생존율은 각각 76.4%, 70.6%, 80.5%, 78.1%로 관찰되었다.

• 한국수산과학회지
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• 제48권4호
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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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• 제39권1호
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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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• 제49권1호
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• pp.43-50
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• 2016

병원급식소의 HACCP 선행요건 수행 실태를 분석하여 병원급식소 특성을 고려한 선행요건관리 개선 방안을 마련하고자 종합병원 규모의 병원 급식소 총 65곳의 급식관리자를 대상으로 설문조사를 실시한 결과, 선행요건 7개 관리영역 중 영업장관리 영역의 수행도는 선행요건 평가판정그룹별로 유의적인 차이가 있었으며, 적합 그룹 (85점 이상)에서 유의적으로 가장 높았다 (p < 0.001). 위생관리 (p < 0.001), 제조 가공 조리 시설 설비관리 (p < 0.001), 용수관리 (p < 0.001), 보관 운송관리 (p < 0.05) 영역의 수행도는 '부적합' 그룹 (70점 미만)이 '적합'이나 '보완' 그룹 (71점 이상 84점 이하)에 비해 유의적으로 낮았다. 조사대상 병원급식소와 영양사의 일반 특성에 따른 선행요건관리 평가 결과에 대한 차이 분석 결과 서울 경인지역이 대구 경북지역에 비해 선행요건 관리가 '적합'으로 평가된 비율이 유의적으로 높았다 (p < 0.01). 또한 선행요건 판정 그룹에 따른 급식시설 구비율은 '적합' 그룹이 '부적합' 그룹에 비해 3조 싱크 (p < 0.01), 대용량 오븐 (p < 0.01), 대형 에어컨 (p < 0.01), 보온고 (p < 0.01), 보냉고 (p < 0.05), 해동전용 냉장고 (p < 0.05), 위생장화 소독고 (p < 0.05) 등 총 7개 항목의 구비율이 유의적으로 높았다. 병원 급식품질 개선을 위해 HACCP을 빠른 시일 내에 효과적으로 적용하기 위해서는 선행요건 관리 항목이 준수되어야 하며, 이에 앞서 병원 급식소의 운영 특성을 고려한 선행요건관리 기준과 HACCP 관리 계획의 수립이 필요하다고 판단된다.

• Clinical and Experimental Reproductive Medicine
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• 제41권3호
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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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• 제46권4호
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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.

• 콘크리트학회논문집
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• 제15권4호
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• pp.529-540
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• 2003

최근 콘크리트 내구성설계에 관한 연구가 활발히 진행되고 있으며, 사용수명 예측이 핵심인 내구성 평가모델 개발이 그 좋은 예이다. 본 연구에서는 콘크리트 구조물의 잔존수명예측 모델을 개발하여 적정시기의 유지보수를 통한 경제적 구조물 사용을 목적으로 하였다. 육지 콘크리트 구조물인 저수지의 콘크리트 구조물 부분을 대상으로 전국 70개 지구를 선정하고, TG/DTA 법과 페놀프탈레인 지시약법으로 중성화를, pH메타법으로 pH 값을 측정하여 탄산칼슘함량 대비 사용연수, pH값, 콘크리트 피복 두께의 관계함수를 각각 유도한 후 가능한 최소의 자료측정으로 잔존수명을 예측할 수 있는 모델을 개발하였다. 개발된 잔존수명예측 모델은 탄산가스등의 고정변수에 의한 실내촉진실험 자료기반 모델과 달리 동결융해작용, 중성화, 철근 부식 등 복합적인 열화작용이 동시에 일어나는 현장의 환경적 영향을 받은 구조물에서 측정한 자료를 기반으로 개발되었다. 이러한 점에서 그 신뢰성을 높게 평가 받을 수 있을 것이며, 시설물 유지관리자에게 적정 보수보강 시점을 제공하여 경제적인 구조물 사용에 도움을 줄 수 있을 것으로 판단된다.