• Title/Summary/Keyword: OPS method

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Comparison of Two Vitrification Methods of In Vitro Development Oocytes Collected from Porcine Antral Follicles Using Open Pulled Straw (OPS) Techniques

  • An, Mihyun;Hong, Daewuk;Son, Dongsoo;Seok, Hobong
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.84-84
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    • 2002
  • The advantages of the OPS techniques(Vajta G et al, Mol Reprod Dev 51: 53-58,1998) give 1) high survival rates of various types of eggs, 2) quick and simple process, 3) inexpensive equipment and reduced chilling injury. The efficiency of IVM/IVF technique in the porcine species is relatively lower than that obtained in other species such as ruminants. Two experiments were designed to investigate the effects of in-vitro fertilization of porcine oocytes matures using different OPS protocol for chilling and warming of vitrification. Porcine oocytes from ovaries collected at abattoir were matured for 44 hours in TCM199 Earle's salt supplemental with pyruvate, pff, L-cysteine, hormones and gentamycin. Oocytes were denuded and fertilized with frozen boar semen by common method. Porcine embryos produced routinely by in-vitro culture system of NCSU23 medium. The vitrification and the warming were conducted by OPS method with the glass micropipette instead of straw vessels and modified the protocol of G.Vajta(1999). In Exp 1, Chilling/Warming:Holding Medium(HM)+EG+DMSO/HM +sucrose Medium(SM) at 39$^{\circ}C$ warm stage. In Exp 2, : PBS+CS+EG+Ficoll+ Trehalose/PBS+Trehalose at 25$^{\circ}C$ stage. Filling, freezing, packing, thawing out and further culturing were performed to follow the basic protocol of G Vajta. During IVM-lVC and post-warming, fertilization parameter and developmental potential were compared to and statistically analysed. It was not significantly different from Exp 1 and Exp 2 but 25$^{\circ}C$ of stage was slightly higher on the morula/blastocyst forming rate and better atmosphere for worker than that at 39$^{\circ}C$ stage.

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Ozone Processing Storage Characteristic of Main Herb Medicines (주요 한약재의 오존처리저장 특성)

  • Woo, Sung-Hun;Lee, Kwang-Sik
    • Journal of the Korean Institute of Illuminating and Electrical Installation Engineers
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    • v.22 no.1
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    • pp.66-72
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    • 2008
  • Recently, many storing methods were applied to keep quality of herb medicines during storage. Among of thent we tried to research a method to store after dissolving remaining harmful bacillus and agricultural chemicals of herb medicines. We confirmed that ozone processing is superior at the sterilization of bacteria and virus from the leading research about ozone generator, and then we made research method to apply with the sterilization and storing processing of herb medicines using ozone (OPS). In this study we chose herb medicines of 4 kinds which have been sold much in the market, evaluated the characteristics. Namely, we used ozone lamp in a storage system and investigated the sterilization characteristic of herb medicines. We executed a quantitative analysis of the general ingredient and the physiological activity ingredient to investigate whether the original ingredient and the quality of herb medicines is preserved, after doing an ozone md storing processing for a long term.

Analysis of Genetic Relationship Among Native Taraxacum and Naturalized Taraxacum species using RAPD (RAPD를 이용한 자생 민들레 종과 귀화 민들레 종간의 연관계 분석)

  • 안영희;박대식;정규환
    • Korean Journal of Environment and Ecology
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    • v.17 no.2
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    • pp.169-176
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    • 2003
  • The genetic relationships between 4 Korean native Taraxacum and 2 naturalized Taraxacum species were analyzed using the random amplified polymorphic DNA (RAPD) method. Because 141 polymorphic bands were generated from 30 random primers selected through the primer screening, it was possible to analyze the genetic relationship among 6 Taraxacum species. In RAED with the primer OPC12, OPD16, OPK16, OPK17, OPK20, OPS1 or OPS8, many specific polymorphic bands have been appeared in each species. Especially RAPD with the primer OPS8, a specific polymorphic band at 564bp was appeared only in the naturalized Taraxacum officinale. Based on RAPD analysis, Korean native Taraxacum and naturalized Taraxacum species are divided into two groups. T. officinale and T. laevigatum are classified into group I which is a naturalized Taraxacum species group, and T. mongolicum, T. hallasanensis, T. ohwianum and T. coreanum are classified into group II which is a Korean native Taraxacum species group. The result from the RAPD method was very similar to the result from the Bootstrap method. From the examination of the physical characteristics of 6 Taraxacum species populated in Korea, flowering period of Taraxacum species in group I are longer than Taraxacum species in group ll, and the direction of involucral bract of Taruxacum species in the group I was also different comparing to the group ll. Because the flowering color, leaf direction, and the specificity of seed germination of T. coreanum were different compared to the other species in the group II, T. coreanum would be genetically divergent and showed the highest dissimilarity index score.

Development of In Vitro Produced Bovine Embryos after Vitrification with Various Containers (다양한 Container로 유리화 동결된 체외생산 소 수정란의 발달에 관한 연구)

  • Lee, Y.J.;Ko, D.H.;H.T., Lee;Chung, K.S.
    • Korean Journal of Animal Reproduction
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    • v.25 no.4
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    • pp.371-379
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    • 2001
  • The objective of this study was to establish an effective cryopreservation method of in vitro-produced bovine embryos. For the vitrification, in virtro-produced embryos at 8-cell, morula and blastocyst stages were exposed to freezing solution containing 5.5 M EG (EG 5.5) for 20 sec, loaded on each containers such as EM grid, OPS and Cryo-loop, and then immediately plunged into liquid nitrogen at -196$^{\circ}C$. Thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-HPBS, each for 1 min, and cultured in CRlaa medium supplemented with 10% FBS. Significant differences in the rates of re-expanded and hatched embryos were not observed among these embryo containers. The total cell number of expanded blastocyst cultured in vitro after vitrification was examined by Hoechst staining. There were no differences between non-vitrified (180.0 $\pm$ 5.4) and vitrified groups (178.0 $\pm$ 7.5). In addition, when the cellular injuries after vitrification were compared by double staining. There were no significant difference in the ratio of live and dead cells between non-vitrified group (176 : 4) and vitrified group (172 : 6). Therefore, these results suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification using various containers, such as EM grid, OPS or Cryo-loop in the presence of EG 5.5 freezing solution.

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Survival and Development of Porcine Embryos Produced in vitro Using Open Pulled Straw Methods (돼지에서 Open Pulled Straw(OPS) 방법에 의해 동결-융해한 수정란의 생존능력)

  • Lee, S.Y.;Yu, J.S.;Sa, S.J.;Park, C.K.
    • Journal of Embryo Transfer
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    • v.21 no.3
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    • pp.255-262
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    • 2006
  • The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) methods on in vitro survival ability of porcine embryos. For in vitro maturation of immature oocytes, the porcine ovaries were collected from local slaughter-house. The cumulus-oocytes complexes were aspirated from 2 to 6 mm follicles. The collected oocytes were cultured for in vitro maturation in NCSU-23 medium with 5 mM hypotaurine, 0.57 mM cysteine, 10% porcine follicle fluid, 10 IU/ml PMSG and 10 IU/ml hCG for $21{\sim}22$ hrs. Then, the oocytes were more cultured $21{\sim}22$ hrs in vitro maturation in medium removed hormones. The frozen-thawed spermatozoa were washed by centrifugation 2 times for 10 min at 1,500 rpm in D-PBS with 5.56 mM glucose, 0.33 mM Na-pyruvate, 100 IU/ml penicillin, $100 {\mu}g/ml$ streptomycin and 4 mg/ml BSA. The fertilization medium used mTBM with 2 mM caffeine and 2 mg/ml BSA and adjusted to a pH of 7.2 to 7.4. The final concentration of spermatozoa was adjusted to $2.5{\times}10^6$cells/ml motile sperm during fertilization in vitro. At 8 hrs after insemination, the oocytes were transferred into NCSU-23 medium with 5.0 mM hypotaurine, 4 mg/ml BSA and 10 ng/ml EGF and cultured for 7 days. When the blastocysts of different stages were frozen-thawed by OPS methods, the proportions of embryos with normal morphology were significantly (p<0.05) higher in embryos frozen-thawed at expanded blastocyst stage (38.9%) than in early blastocyst stage (28.3%). On the other hand, the proportions of embryos damaged after frozen-thawing were significantly (p<0.05) higher in embryos frozen at early blastocyst stages than in expanded blastocyst stage. In another experiment, the normal embryos morphology after frozen- thawing were further cultured for 48 hrs. After culture, the proportions of embryos hatched were 6.7, 20.0 and 33.3% for embryos frozen-thawed at early blastocyst, mid-blastocyst and expanded blastocyst stages. These finding indicate the possible broader application for OPS methods, as frozen-thawed embryos may be accompanied by developmental stage according to requirements of the survival ability after freezing of blastocyst stage in the pig.

A Study of Application Development Method for Improving Productivity on Cloud Native Environment (Cloud Native환경에서의 생산성 향상을 위한 어플리케이션 개발 방법 연구)

  • Kim, Jung-Bo;Kim, Jung-In
    • Journal of Korea Multimedia Society
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    • v.23 no.2
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    • pp.328-342
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    • 2020
  • As the cloud-based ICT(Information & Communication Technology) infrastructure matures, the existing monolithic software development method is evolving into a micro-service structure based on cloud native computing. To develop and operate the services efficiently under the cloud native environment, DevOps-based application development plans through MSA(Micro Service Architecture) design based are essential. A cloud native environment is an approach to developing and running applications that take advantage of cloud computing models such as automation of source distribution, container-based virtualization, application scalability, resource efficiency, and flexible maintenance through object independence. To implement this approach, the utilization of key elements such as DevOps, continuous delivery, micro service, and containers is essential, but there are not enough previous studies on case analyses or application methods of these key elements. Therefore, in this paper, we analyze the cases of application development in cloud native environment and propose the optimized application development process and development method through small and medium-sized SI projects.

Bayesian estimation of the Korea professional baseball players' hitting ability based on the batting average (한국프로야구 선수들의 타율에 기반된 타격 능력의 베이지안 추정)

  • Cho, Yong Ju;Lee, Kwang Ho
    • Journal of the Korean Data and Information Science Society
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    • v.26 no.1
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    • pp.197-207
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    • 2015
  • In baseball game, the hitting ability of batter is frequently assessed by a batting average, a run batted in, a home run, a run scored, an on-base percentage, etc. Recently, more comprehensive indicators such as OPS, ISO, SECA, TA, RC and XR are often used. But, these measures generally shows large deviations since they are calculated from the data for a certain period of time, and they are not an estimate of a population parameter, either. In this paper, we will presume the pure hitting ability of the korea professional baseball players as a parameter which is depend upon at bat. We will estimate the parameter by using the Bayesian method.

Effects of Cryoprotectant, Warming Solution and Removal of Lipid on Viability of Porcine Nuclear Transfer Embryos Vitrified by Open Pulled Straw Method

  • Cong, Pei-Qing;Song, Eun-Sook;Kim, Eui-Sook;Li, Zhao-Hua;Zhang, Yong-Hua;Lee, Jang-Mi;Yi, Young-Joo;Park, Chang-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.103-108
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    • 2007
  • This study was carried out to investigate the effects of cryoprotectants, warming solution and removal of lipid on open pulled straw vitrification (OPS) method of porcine embryos produced by nuclear transfer (NT) of fetal fibroblasts. All solutions used during vitrification were prepared with holding medium consisting of 25 mM Hepes buffered TCM199 medium containing 20% fetal bovine serum (FBS) at $38.5^{\circ}C$. The blastocysts derived from NT with or without lipid were vitrified in each medium of different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG). Also, blastocysts after cryopreservation were warmed into different concentrations of sucrose in warming solution. The optimal concentrations of cryoprotectants in vitrification solution were 10% DMSO + 10% EG in vitrification solution 1 (VS1) and 20% DMSO + 20% EG in vitrification solution 2 (VS2). The optimal concentrations of sucrose were 0.3 M sucrose in warming solution 1 (WS1) and 0.15 M sucrose in warming solution 2 (WS2). lipid removal from oocytes before NT enhanced the viability of NT embryos after vitrification. Our results show that use of the OPS method in conjunction with lipid removal provides effective cryopreservation of porcine nuclear transfer embryos.

Study on the Development of Efficient Vitrification of Human Blastocysts (인간포배기 배아의 효과적인 유리화 동결법의 개발을 위한 연구)

  • Lee, Sang-Min;Lee, Ju-Hee;Lee, Sang-Won;Lee, Seoung-Min;Yoon, San-Hyun;Lim, Jin-Ho;Park, Huem-Dai;Lee, Seong-Goo
    • Clinical and Experimental Reproductive Medicine
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    • v.30 no.3
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    • pp.241-248
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    • 2003
  • Objective: The purpose of this study was to evaluate the survival rate of vitrified blastocyst according to the freezing vessels, equilibration time in cryoprotectant and artificial dehydration method. Methods: Human blastocysts were vitrified after loading onto the plastic straw, open-pulled straw (OPS), electron microscopy grid (EM grid) for 1.5 min or 3 min. They also were directly plunged into LN2 within 30sec. For artificial shrinkage of blastocysts, 36 gauge fine needle was pushed at the cellular junction of the trophectoderm into the blstocoele cavity until it shrank without damage of inner cell mass. Results: The survival rate of vitrified blastocysts on plastic straw, OPS, EM grid as freezing vessels were 26.7, 13.0 and 60.5%, respectively. The survival rate of EM grid was significantly higher than that of plastic straw and OPS (p<0.05). For 1.5 min equilibrium, the survival rates of early blastocyst (EB), middle blastocyst (MB) and late blastocyst (LB) were 64.4, 81.0, and 20.0% respectively. For 3 min equilibrium, the survival rates of EB, MB, and LB were 69.9, 50.0 and 57.5% respectively. The survival rates of EB and MB were significantly higher than that of LB in 1.5 min equilibrium group (p<0.05), however, the significance was not observed in 3 min equilibrium groups. In cytoplasmic shrinkage before vitrification, the survival rates of EB, MB and LB were 92.9, 100 and 75.9% respectively. The survival rate of MB was significantly higher than that of LB (p<0.05). The survival rates of vitrified blastocysts by artificial dehydration and slow-frozen blastocysts were not significantly different as 88.9 and 66.7%, respectively. Conclusion: This study showed that the vitrification of human blastocysts using EM grid and artificial dehydration is an effective method. Therefore, these methods would be an useful techniques for blastocyst cryopreservation.

Survival Ability of Porcine Oocytes Frozen-Thawed by Open Pulled Straw Method (Open Pulled Straw 방법에 의해 동결-융해된 돼지난자의 생존능력)

  • 김세웅;박춘근;정희태;양부근;김정익
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.117-125
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    • 2001
  • Vitrification of oocytes has been applied recently fur pigs, but remains elusive. The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine oocytes. When immature follicular oocytes frozen-thawed were cultured for in vitro maturation, maturation rates to metaphase-II stage were higher in oocytes with (25%) than without (15%) cumulus cells. After In vitro fertilization of oocytes frozen-thawed, the maturation rates were also significantly (P<0.05) higher in oocytes with (41%) that than without (17%) cumulus cells. However, the penetration rates were higher in oocytes without (19%) that than with (9%) cumulus. In another experiment, porcine oocytes matured in vitro were frozen and thawed for in vitro fertilization. The penetration rates were higher than in oocytes without (35%) that than with (26%) cumulus cells. However, the proportions of oocytes dead after in vitro fertilization were significantly (P<0.05) higher in oocytes with that than without cumulus cells. On the other hand, the rates of penetration and dead oocytes at 6 h after in vitro fertilization were not significant differences between oocytes with and without cumulus cells. However, the proportions of dead oocytes with (18%) and without (16%) cumulus cells were higher than in oocytes of control group (0%). These finding indicated the possible broader application for OPS, as they demonstrated that the maturation and fertilization in vitro by frozen-thawed oocytes may be accompained by cumulus cells and culture periods according to the requirements of the survival ability after freezing of mature and immature oocytes in pigs.

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