• Title/Summary/Keyword: Freezing Method

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Cryopreservation of the Human Adult Ovarian Cortical Tissues by Vitrification (여성의 난소 피질조직의 초자화 냉동보존)

  • Lee, K.A.;Lee, S.H.;Ha, S.D.;Yoon, S.J.;Ko, J.J.;Lee, W.S.;Yoon, T.K.;Cha, K.Y.
    • Clinical and Experimental Reproductive Medicine
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    • v.26 no.2
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    • pp.251-256
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    • 1999
  • The present study was conducted to evaluate whether vitrification could be used for ovarian tissue preservation. The important issue here is that the vitrification is very simple, easy, and economical compared to the conventional cryopreserving method that using automatic freezing instrument. Human ovarian cortical tissues were cryopreserved by vitrification with 5.5 M ethylene glycol and 1.0 M sucrose as cryoprotectant. Three points of temperature ($4^{\circ}C$, room temperature, and $37^{\circ}C$) and two points of duration (5 or 10 minutes) for cryoprotectant treatment were examined to determine the best condition for vitrification of the human ovarian cortical tissues. After thawing, viability of the isolated primordial follicles was examined by dye-exclusion method. Histological appearance of tissues before and after the cryopreservation was evaluated. There was no toxic effect of the 5.5 M ethylene glycol on the primordial follicles. However, when the tissues were treated with cryoprotectant at $37^{\circ}C$ for 10 minutes and exposed to liquid nitrogen, it seems likely that there is certain deleterious effects on the viability of the primordial follicles. The highest viability of the primordial follicles was obtained with the treatment of cryoprotectant at room temperature for 10 minutes. Follicles and oocytes survived after freezing and thawing had the similar normal shapes as was seen in the specimens before cryopreservation. It would be useful to apply vitrification in establishing ovarian tissue banking for clinical purposes.

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Influencing Factors on Freezing Characteristics of Frost Susceptible Soil Based on Sensitivity Analysis (민감도 분석을 기반으로 한 시료의 동결 특성에 미치는 영향인자 분석)

  • Go, Gyu-Hyun;Lee, Jangguen;Kim, Minseop
    • Journal of the Korean Geotechnical Society
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    • v.36 no.8
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    • pp.49-60
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    • 2020
  • A fully coupled thermo-hydro-mechanical model is established to evaluate frost heave behaviour of saturated frost-susceptible soils. The method is based on mass conservation, energy conservation, and force equilibrium equations, which are fully coupled with each other. These equations consider various physical phenomena during one-dimensional soil freezing such as latent heat of phase change, thermal conductivity changes, pore water migration, and the accompanying mechanical deformation. Using the thermo-hydro-mechanical model, a sensitivity analysis study is conducted to examine the effects of the geotechnical parameters and external conditions on the amount of frost heave and frost heaving rate. According to the results of the sensitivity analysis, initial void ratio significantly affects each objective as an individual parameter, whereas soil particle thermal conductivity and temperature gradient affect frost heave behaviour to a greater degree when applied simultaneously. The factors considered in this study are the main factors affecting the frost heaving amount and rate, which may be used to determine the frostbite sensitivity of a new sample.

Frost Heave Force of Ground and Countermeasure for Damage of Structures (지반의 동상력과 구조물의 피해대책)

  • Rui, Da-Hu;Teruyuki, Suzuki;Kim, Young-Su
    • Journal of the Korean Geotechnical Society
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    • v.23 no.5
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    • pp.43-51
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    • 2007
  • Frost action may cause extensive damage to building, structures, roads, railways and utility lines in seasonal frost. The research about frost heave of natural ground has been considerably performed. In late years various structures have become complicated with the development of social infrastructure maintenance. Therefore countermeasure to frost heave becomes a matter of great importance from a new viewpoint. This study was aimed at catching natural ground frost heaving force quantitatively. Frost heaving forces on circular steel plates which were set on ground surface were measured in field test. The frost heaving forces arise at freezing front propagates to the structures through frozen soil layer. Besides, a full scale model of multi-anchored retaining wall was installed in field, and the freezing lines, frost heave pressure to act on a wall block, and so on were measured. Finally, the position and shape of frost line were estimated by using numerical simulation and a method to determine replacement range was suggested with soil properties and weather data.

Numerical Study on the Production of Methane Hydrate by Depressurization Method (감압법을 이용한 메탄하이드레이트 생산에 대한 수치적 연구)

  • Kim, Jin-Hong;Chun, Won-Gee;Kim, Nam-Jin
    • 한국신재생에너지학회:학술대회논문집
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    • 2007.06a
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    • pp.519-523
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    • 2007
  • Gas(or methane) hydrates are solid solutions when water molecules are linked through hydrogen bonding and create host lattice cavities that can enclose a large variety of guest gas molecules. The natural gas hydrate crystal may exist at low temperature above the normal freezing point of water and high pressure greater than about 30 bars. A lot of quantities of natural gas hydrates exists in the earth and many production schemes are being studied. In the present investigation, depressurization method was considered to predict the production of gas and the simulation of the two phase flow - gas and water - in porous media is being carried out. The simulation show about the fluid flow in porous media have a variety of applications in industry. Results provide the appearance of gas and water production, the pressure profile, the saturation of gas/ water/ hydrates profiles and the location of the pressure front.

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Experimental Study for Development of Air Eject Defrost Equipment (공기분사식 제상장치 개발에 관한 실험적 연구)

  • Han, In-Geun;Kim, Chang-Yeong;Kim, Jae-Dol;Yun, Jeong-In
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.25 no.3
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    • pp.277-284
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    • 2001
  • One of the problems in a refrigerator operation is the frost formation on a cold surface of the evaporator. The frost layer is formed by the sublimation of water vapor when the surface temperature is below the freezing point. This frost layer is usually porous and formed on the cold surface of the evaporator. The frost layer on the surface of a evaporator will make side effect such as thermal resistance. However, these important factors have not been used in determining the defrosting period. Therefore, the proper defrosting operation period based on the new defrosting method is very important, and make a comparison between conventional method like electric defrost and new method in compression work, evaporation pressure, evaporation temperature.

Limb Salvage Operation with Recycled Autogenous Bone Graft (자가골 재이식술을 이용한 사지 구제술)

  • Rhee, Seung-Koo;Kang, Yong-Koo;Suh, Yoo-Joon;Yoo, Jong-Min;Jung, In-Ho
    • The Journal of the Korean bone and joint tumor society
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    • v.10 no.2
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    • pp.96-106
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    • 2004
  • Purpose: To determine the usefulness of limb salvage operation with recycled autogenous bone graft in musculoskeletal malignant tumors. Materials and Methods: Twenty nine cases, who underwent limb salvage operation with recycled autogenous bone graft for the treatment of musculoskeletal malignant tumor between February 1990 and January 2003, were included. There were 18 males and 11 females and the mean age was 33 years (range, 10 to 65 years). The mean follow-up period was 51.8 months (range, 18 to 117 months). The Enneking stage was IIA in 10 cases and IIB in 19 cases. The recycling method of autogenous bone was deep freezing in 6 cases, autoclaving in 11 cases, pasteurization in 7 cases and the composite of autoclaving and vascularized fibular graft in 5 cases. The union of junctional site was evaluated radiologically and the functional results was analyzed by the grading systems of the International Symposium On Limb Salvages (ISOLS). Results: The mean union time was 7.2 months (range, 3 to 15 months). The union took 5.8 months (range, 4 to 8 months) in deep freezing, 9.7 months (range, 6 to 15 months) in autoclaving, 5.9 months (range, 4 to 8 months) in pasteurization, and 5 months (range, 4 to 8 months) in the composite of autoclaving and vascularized fibular graft. The mean functional evaluation percentage was 76.8% (range, 40 to 90%). It was 65.8% (range, 40 to 85%) in deep freezing, 76.6% (range, 40 to 90%) in autoclaving, 81.6% (range, 70 to 90%) in pasteurization, and 83.4% (range, 75 to 90%) in the composite of autoclaving and vascularized fibular graft. There were 6 cases of complications including 1 case of local recurrence, lung metastasis, infection, fracture, respectively and 2 cases of nonunion. Conclusion: The limb salvage operation with recycled autogenous bone graft is a useful treatment method for the musculoskeletal malignant tumors. Particularly, autoclaving is the most reliable sterilization method. The vascularized fibular graft can compensate decreased osteoinductivity and mechanical strength of recycled bone. So, the composite of autoclaving and vascularized bone graft seems to be a favorable treatment method for high grade malignant musculoskeletal tumors.

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Correction of Mean and Extreme Temperature Simulation over South Korea Using a Trend-preserving Bias Correction Method (변동경향을 보존하는 편의보정기법을 이용한 우리나라의 평균 및 극한기온 모의결과 보정)

  • Jung, Hyun-Chae;Suh, Myoung-Seok
    • Atmosphere
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    • v.25 no.2
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    • pp.205-219
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    • 2015
  • In this study, the simulation results of temperature by regional climate model (Reg- CM4) over South Korea were corrected by Hempel et al. (2013)'s method (Hempel method), and evaluated with the observation data of 50 stations from Korea Meteorological Administration. Among the 30 years (1981~2010) of simulation data, 20 years (1981~2000) of simulation data were used as a training data, and the remnant 10 years (2001~2010) data were used for the evaluation of correction. In general, the Hempel method and parametric quantile mapping show a reasonable correction both in mean and extreme climate of temperature. As the results, the systematic underestimation of mean temperature was greatly reduced after bias correction by Hempel method. And the overestimation of extreme climate, such as the number of TN5% and freezing day, was significantly recovered. In addition to that, the Hempel method better preserved the temporal trend of simulated temperature than other bias correction methods, such as the quantile mapping. However, the overcorrection of the extreme climate related to the upper quantile, such as TX5% and hot days, resulted in the exaggeration of the simulation errors. In general, the Hempel method can reduce the systematic biases embedded in the simulation results preserving the temporal trend but it tends to overcorrect the non-linear biases, in particular, extreme climate related to the upper percentile.

The Experimental Study on Cryopreservation of Mouse Embryo (생쥐배아의 동결보존에 관한 실험적 연구)

  • Lee, Yu-Il;Kwon, Young-Sook;Park, Hyun-Jeong
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.1
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    • pp.55-63
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    • 2001
  • Objectives: This study was carried out to evaluate the effects of embryonic stage, cryoprotectant, and freezing-thawing method on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology (ART). Materials and Methods: Two to eight cell embryos were obtained from oviducts of mated $F_1$ hybrid female mice superovulated by pregnant mare's serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Two-step 1,2-propanediol (PROH), dimethylsulfoxide (DMSO) and 4-step PROH DMSO were used as cryoprotectant and dehydration and rehydration method of embryos, and slow-cooling or rapid-cooling method was used as frozen program. The survival rates of embryos were measured after thawing and rehydration, and the developmental rates of embryos were compared and observed during culturing embryos for 24, 48, 72, 96 hrs. Results: As for the survival and development rates of embryos according to embryonic stage, the survival rate of 2 cell stage in PROH and DMSO was significantly higher than 4-8 cell (64.5% versus 62.1 %,79.7% versus 73.2%) (p<0.01, p<0.01), but the development rates of 4-8 cell embryos in PROH and DMSO were significantly higher than 2 cell embryos for whole culture period (p<0.01) and the development rates of 4-8 cell embryos in PROH were significantly higher than 2 cell embryos in DMSO (p<0.01). As for the survival and development rates of embryos according to cryoprotectant, the survival rate of 2 cell embryo in DMSO was significantly higher than that in PROH (74.4% versus 64.5%) (p<0.01), whereas the development rate of embryos was not differ till 24 hrs. The developmen1 rate from morular to hatching blastocyst, however, was significantly higher in PROH than in DMSO during 48 hr (p<0.01). The survival rate of 4-8 cell embryo was 62.1% in PROH and 73.2% in DMSO. The development rates of embryo in PROH were significantly higher for whole culture periods (p<0.01, 0.05). In respect to the effect of freezing and thawing program on the survival and development rates of embryos, method of slow cooling and rapid thawing was more effective than that of rapid cooling and rapid thawing. Conclusions: The survival rate of embryo in 2 cell stage was higher than in 4-8 cell stage, and PROH appears more effective cryoprotectant than DMSO because PROH showed better development rates of embryos in 2 and 4-8 cell stage. Moreover, slow cooling and rapid thawing method was considered as the best cryopreservation program.

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Effect of Ethylene Glycol(EG) and Propylene Glycol(PG) on the Viability of Frozen-thawed Primordial Germ Cells(PGCs) on Korean Native Chicken(Ogye) by Vitrification (한국재래닭(오계)의 유리화 동결 시 생존율에 미치는 Ethylene Glycol(EG)과 Propylene Glycol(PG)의 영향)

  • Kim, Hyun;Kim, Dong Hun;Han, Jae Yong;Choi, Sung Bok;Ko, Yeoung Gyu;Do, Yoon Jung;Seong, Hwan-Hoo;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.197-205
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    • 2013
  • This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and 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 on improving the production of germline chimeras.

Development of Weight Type Rounded Snow Plate (중량식 원형 적설판 개발에 관한 연구)

  • Lee, Bu-Yong;Kim, Hyun-Chul
    • Atmosphere
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    • v.19 no.1
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    • pp.1-8
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    • 2009
  • We need water equivalent unit data of snowfall for the purpose of forecast and hydrology related research area. This study developed new method of automatic recording snowfall as weight unit with circle type plate using stain-gauge loadcell. Field test of instrument carried out at Daegwallyeong Obs. Station from 20 to 23 Jan. 2008 during heavy snowfall. There is 74.2cm snow depth and 54.6mm precipitation by Daegwallyeong Obs. Station. But the instrument of this study recorded 71.0mm of precipitation amount. Because of different observation method can cause more 15.4mm than Daegwallyeong Obs. Station. But this study gives the possibility of observation of new snow fall measurement under freezing conditions of snow. From the observation data the density of snowfall calculated from 0.09 to $1015g/cm_3$ from the observation period. And have a good relations between manual observation and automatic observation data from this study instrument with slope of 1.35 to 1.39.