• Title/Summary/Keyword: Liquid Nitrogen Storage

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Optimization of the cryopreserved condition for utilization of GPCR frozen cells (GPCR 냉동보관 세포의 활용을 위한 냉동조건의 최적화 연구)

  • Noh, Hyojin;Lee, Sunghou
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.16 no.2
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    • pp.1200-1206
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    • 2015
  • The major target for drug discovery, G-protein coupled receptor (GPCR) is involved in many physiological activities and related to various diseases and disorders. Among experimental techniques relating to the GPCR drug discovery process, various cell-based screening methods are influenced by cell conditions used in the overall process. Recently, the utilization of frozen cells is suggested in terms of reducing data variation and cost-effectiveness. The aim of this study is to evaluate various conditions in cell freezing such as temperature conditions and storage terms. The stable cell lines for calcium sensing receptor and urotensin receptor were established followed by storing cultured cells at $-80^{\circ}C$ up to 4 weeks. To compare with cell stored at liquid nitrogen, agonist and antagonist responses were recorded based on the luminescence detection by the calcium induced photoprotein activation. Cell signals were reduced as the storage period was increased without the changes in $EC_{50}$ and $IC_{50}$ values $EC_{50}:3.46{\pm}1.36mM$, $IC_{50}:0.49{\pm}0.15{\mu}M$). In case of cells stored in liquid nitrogen, cell responses were decreased comparing to those in live cells, however changes by storage periods and significant variations of $EC_{50}/IC_{50}$ values were not detected. The decrease of cell signals in various frozen cells may be due to the increase of cell damages. From these results, the best way for a long-term cryopreservation is the use of liquid nitrogen condition, and for the purpose of short-term storage within a month, $-80^{\circ}C$ storage condition can be possible to adopt. As a conclusion, the active implementation of frozen cells may contribute to decrease variations of experimental data during the initial cell-based screening process.

Short-Term Storage and Cryopreservation of Abalone (Haliotis discus hannai) Sperm

  • Kang, K.H.;Kho, K.H.;Chen, Z.T.;Zhang, Z.F.;Chang, Y.J.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.74-74
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    • 2003
  • In present study, attempts were made to preserve abalone (Haliotis discus hannai) sperm in liquid form at low temperature, to evaluate the effect of various diluents in short-term storage on sperm, and cryopreservation procedures were optimized for the cryoprotectants as well as freezing rates, in terms of the motility and survival rate, and the ultrastructural changes of sperm after short-term storage and cryopreservation were observed. The abalone sperm reached maximum motility until about 4min after activation. The motility was constant for about 16min, after which it dropped gradually, and about 50min later all motility ceased. Threshold activation of sperm was found in 40% artificial seawater (ASW), and motility increased as the concentration of ASW increased. In Hanks balanced salt solution without calcium (Ca-Free HBSS, 300 and 400 mOsmol/kg) and 10%, 20%, and 30% ASW the sperm was immotile, and motility once again restored incompletely only in HBSS of 300 and 400 mOsmol/kg, 20% and 30% ASW after 100% ASW was added. Sperm motility was extended following 20 days of cold storage only in 70% and 100% ASW. A high motility index of 3.5-4.5 was observed for the first 8 days in 70% and 80% ASW. In other diluents sperm motility was constant less than 10 days, and the motility index was obviously lower than that of sperm in 70% and 100% ASW. After 20 days of cold storage survival rates of 10.2%-20.7% were obtained in ASW and 300 mOsmol/kg HBSS, and that in 400 HBSS (65.3%) was significantly higher than others. The constant period of sperm motility stored in 70% ASW was longer obviously than that in 100% ASW after 6 days of storage, and the time to maximum motility of sperm stored in 70% increased gradually, while the difference in which of sperm in 100% ASW was not significant. The sperm plunged into liquid nitrogen all died except that sperm using 15% glycerol as cryoprotectant restored 10.4% of motility. The highest motility index (3.4) was obtained with 5% glycerol and freezing procedure: $50^{\circ}C$/min from $20^{\circ}C$ to $-80^{\circ}C$.

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Effects of Enzyme Complex on Odor Emission from Swine Slurry and Swine Buildings (효소복합체가 양돈슬러리 및 돈사 악취발생에 미치는 영향)

  • Jung, K.H.;Han, J.C.;Kwack, S.J.;Jung, J.D.;Lee, J.W.;Kim, D.H.
    • Journal of Animal Environmental Science
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    • v.14 no.1
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    • pp.15-22
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    • 2008
  • This study was carried out to investigate the effect of enzyme complex on odor emission from swine slurry and the process of making swine liquid manure. Ammonia and hydrogen sulfide concentrations were significantly decreased by using the enzyme complex of liter per ton level of liquid swine slurry in the manure storage tank according to the time. Characteristics of liquid swine slurry were affected by the enzyme complex, total nitrogen and ammonia nitrogen contents were reduced compare with control. Ammonia and hydrogen sulfide concentrations in the finishing pig building and offensive odor compound on the boundary line of swine farm were significantly decreased by spraying in swine finishing building. In conclusion, the results obtained from this study suggest that using the enzyme complex of liter per ton level of liquid swine slurry for making liquid swine manure may improve the quality of swine liquid fertilizer and reduce odor emission. Also farm scale enzyme complex treatment may improve air quality in finishing pig building and deduce offensive odor compound of swine farm.

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Preconditioning for Cryopreservation of in vitro Grown Bulblets of Lily using Droplet-Vitrification

  • Song, Jae-young;Lee, Young-yi;Yi, Jung-yoon;Lee, Jung-ro;Yoon, Mun-sup
    • Korean Journal of Plant Resources
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    • v.33 no.6
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    • pp.689-695
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    • 2020
  • This study was conducted to improve and supplement the system of cryopreservation for adventitious bulbs induced by tissue cultured bulb-scales of lily (Lilium spp.) cvs. 'Milky way'. The explants, bulblets and bulb-scale-bulblets, were treated to low temperature (4℃) for 7 days prior to the pre-culture. The adventitious bulbs were pre-cultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3 and 0.7M). The pre-cultured adventitious bulbs were treated to loading solution (LS1 or LS2, C4 or C6) containing 35% of PVS3 (LS1, C4) or 40% of PVS3 (LS2, C6) for 40 min and exposed to dehydration solution (PVS3, B1) containing 50% glycerol and 50% sucrose for 60 min at 25℃. The adventitious bulbs were moved onto droplets containing 3 µl PVS3 on sterilized aluminum foils, and then soaked into liquid nitrogen (LN) for 60 min. The result of highest regrowth rate as 65.7% was obtained in cold treatment (4℃), osmoprotected with LS1 solution, and cultured in PCM3 medium by using bulb-scale-bulblet for cryopreservation. This result shows that droplet-vitrification could be used as a promising method for long-term storage of lily genetic resource.

Preconditioning for Cryopreservation of in vitro Grown Bulblets of Lily using Droplet-Vitrification

  • Jae-young Song;Jinjoo Bae;Young-yi Lee;Jung-yoon Yi;Jung-ro Lee;Mun-sup Yoon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.35-35
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    • 2021
  • This study was conducted to improve and supplement the system of cryopreservation for adventitious bulbs induced by tissue cultured bulb-scales of lily (Lilium spp.) cvs. 'MilkyWay'. The explants, bulblets and bulb-scale-bulblets, were treated to low temperature (4℃) for 7 days prior to the pre-culture. The adventitious bulbs were pre-cultured in Murashige and Skoog (MS) liquid medium supplemented with sucrose (0.3 and 0.7M). The pre-cultured adventitious bulbs were treated to loading solution (LS1 or LS2, C4 or C6) containing 35% of PVS3 (LS1, C4) or 40% of PVS3 (LS2, C6) for 40 min and exposed to dehydration solution (PVS3, B1) containing 50% glycerol and 50% sucrose for 60 min at 25℃. The adventitious bulbs were moved onto droplets containing 3 ㎕ PVS3 on sterilized aluminum foils, and then soaked into liquid nitrogen (LN) for 60 min. The result of highest regrowth rate as 65.7% was obtained in cold treatment (4℃), osmoprotected with LS1 solution, and cultured in PCM3 medium by using bulb-scale-bulblet for cryopreservation. This result shows that droplet-vitrification could be used as a promising method for long-term storage of lily genetic resource.

Numerical Investigation on the Urea Melting Characteristics with Coolant and Electric Heaters (냉각수 및 전기 가열 방식에 따른 요소수 해동 특성에 관한 수치해석 연구)

  • Lee, Seung Yeop;Kim, Man Young
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.40 no.1
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    • pp.1-7
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    • 2016
  • A Urea-SCR(Selective Catalytic Reactor) system, which converts nitrogen oxides into nitrogen and water in the presence of a reducing agent, creates a major exhaust gas aftertreatment system for NOx reduction among other compounds. With regard to vehicle applications, a urea solution was chosen based on its eutectic composition of a 32.5wt% urea-water solution. An important advantage of this eutectic composition is that its melting point of $-11.7^{\circ}C$ is sufficiently low to avoid solidification in cold environments. However, the storage tanks must be heated separately in case of low ambient temperature levels to ensure a sufficient amount of liquid is available during scheduled start ups. In this study, therefore, a numerical investigation of three-dimensional unsteady heating problems analyzed to understand the melting processes and heat transfer characteristics including liquid volume fraction, temperature distributions, and temperature profiles. The investigations were performed using Fluent 6.3 commercial software that modeled coolant and electric heater models based on a urea solution. It is shown that the melting performance with the electric heater is higher than a coolant heater and is more efficient.

Cryopreservation of Filefish (Thamnaconus septentrionalis) Sperm

  • K.H. Kang;Z.T. Chen;K.H. Kho;Z.F. Zhang;Kim, J.M.;Kim, Y.H.
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 2003.10a
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    • pp.73-73
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    • 2003
  • The present study examined the possibility of long term storage, by cryopreservation in liquid nitrogen, of the sperm of Filefish (Thamnaconus septentrionalis), and the changes in motility, survival rate and ultrastructure of the sperm after freezing and thawing. The sperm was collected by stripping and stored on ice until experiments. For selection of the immobilizing solution, diluted artificial seawater (ASW) of 20, 30 and 40% were tested. The sperm motility was significantly inhibited in 30% ASW, and restored entirely after 100% ASW was added again. Two cryoprotectants, dimethyl sulfoxide ($Me_2$SO) and glycerol, were added to 30% ASW to formulate the extenders at the concentrations between 5 to 20% by volume for freezing. The sperm was diluted at the ratio of 1 :6 with the extenders, inserted into 0.5ml plastic straws and frozen at a freezing rate of $50^{\circ}C$/min to $-100^{\circ}C$ after equilibration for 10 min at room temperature, followed by plunging into liquid nitrogen. The straws were thawed in a $30^{\circ}C$ water bath for 15 sec. The highest post-thawed sperm motility and survival rate were obtained with 5% glycerol Afterward, the effect of different freezing rates was examined using 5% glycerol as a cryoprotectant, and the rate of $20^{circ}C/min to $-80^{\circ}C$ showed the best result Some ultrastructural changes of sperm, such as the detachment of plasmatic and nuclear membranes, destruction of mitochondria, were observed after cryopreservation. Morphological normality of the sperm in 5% glycerol frozen at the ratio of 1$0^{\circ}C$/min to $-80^{\circ}C$ was better than that of others.

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Cryopreservation of Forest Tree Seeds: A Mini-Review

  • Gantait, Saikat;Kundu, Suprabuddha;Wani, Shabir Hussain;Das, Prakash Kanti
    • Journal of Forest and Environmental Science
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    • v.32 no.3
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    • pp.311-322
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    • 2016
  • Since forest trees form the basis of forest ecosystem, their prolong subsistence is crucial for various flora and fauna. The foremost challenges to sustain the forest ecosystem comprise of the declining forest tree population accompanied with structural changes due to afforestation and exploitation of forest area, environment changes, pests, pollution, and introgressive hybridization. For ex situ conservation approach, in vitro techniques encompass basic role for conserving tree genetic resources, predominantly where natural propagules like recalcitrant seed might not be appropriate for long-term conservation. The practice includes restricted growth techniques, conventional micropropagation, production and storage of synthetic seeds, and cryopreservation. Even though these practices have been applied chiefly to herbaceous species, but recently, woody species were also focused upon. Key conceptions, challenges and techniques for forest tree seed conservation are discussed briefly in this review with special emphasis on some successful cryopreservation approaches for long-term storage.

Squeeze Film Dampers for High Temperature Superconducting Radial Magnetic Bearings

  • Na, Uhn-Joo;Choi, Sang-Kyu;Sung, Tae-Hyun;Han, Young-Hee
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2002.11b
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    • pp.399-403
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    • 2002
  • Squeeze film dampers (SFDs) are designed and analyzed for radial superconducting bearings. The designed SFDs are mounted on the superconductors submerged in liquid nitrogen such that the dampers should supply additional damping to the relatively underdamped superconducting bearing support . Basic theory of SFD assembled with superconductors is introduced. Rotordynamic simulations are provided to support the feasibility of the superconducting bearings mounted on SFDs for a horizontal flywheel energy storage system.

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Development of the Fabrication Technology of High Tc Superconductor for Electrical Energy Storage (전기 에너지 저장을 위한 초전도 나노 합성 기술)

  • Lee, Sang-Heon
    • The Transactions of the Korean Institute of Electrical Engineers C
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    • v.55 no.9
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    • pp.442-445
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    • 2006
  • In order to realize the commercial application of HTSC materials, it is necessary to develop the fabrication process of high Tc oxide superconductor materials with desired shape and for practical application and high critical current density as well as good mechanical strength which can withstand high lorenz force generated at high magnetic field. Much studies have been concentrated to develop the fabrication technique for high critical current density but still there are a lot of gap which should be overcome for large scale application of HTSC materials at liquid nitrogen temperature. Recently some new fabrication techniques have been developed for YBaCuO bulk superconductor with high mechanical strength and critical current density. In this project, the establishment of fabrication condition and additive effects of second elements were examined so as to improve the related properties to the practical use of YBaCuO superconductor, and we reported the production of the YBaCuO high Tc superconductor by the pyrolysis method.