• Title/Summary/Keyword: freezing condition

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Cryopreservation of Bovine IVM/IVF/IVC Hatched Blastocysts (체외생산된 소 완전탈출 배반포기배의 동결보존)

  • Lee, K.S.;Kim, E.Y.;Yi, B.K.;Nam, H.K.;Yoon, S.H.;Park, S.P.;Lim, J.H.
    • Korean Journal of Animal Reproduction
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    • v.23 no.2
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    • pp.141-148
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    • 1999
  • This study was to test whether the viability of bovine hatched blastocysts (HBs) can be maintained after vitrification and thawing. The HBs were produced in vitro at Day 9 and Day 10 after IVF, and they were classified to small (S-HBs; ø$\leq$300 ${\mu}{\textrm}{m}$) and large(L-HBs; ø>300 ${\mu}{\textrm}{m}$) on the basis of embryo diameter using eyepiece micrometer. As freezing solution, we used EFS35 which consisted of 35% ethylene glycol (EG), 18% ficoll, 0.3 M sucrose and 10% FBS added in mDPBS. Vitrification was taken by two-step method, the HBs were equilibrated in 10% EG for 5 minutes and then shortly exposed in EFS35 and plunged into L$N_2$for 30~45 sec. After thawing, the survival rates were assessed by the re-expansion of the blastocoel during 2 h and 16 h of culture. The results obtained in these experiments were summarized as follows; 1) When the blastocysts(40.8%) recovered at Day 8 after IVF were further cultured for 24 h(Day 9 after IVF) and 48 h(Day 10 after IVF), the rates of HBs were 20.5% and 6.7%, respectively. Also, the total cell number of HBs on Day 9 was significantly higher than that of HBs on Day 10 (p<0.01). 2) When the effects of freezing solution to the survival of Day 9 L-HBs were examined, the rate of vitrified group (75.7%) was significantly lower than 100% of control and exposed group(p<0.05). 3) When the survival rates of vitrified HBs according to size and developmental age were examined, the data of L-HBs (75.5%) and S-HBs(63.6%) on Day 9 were slightly higher than those of L-HBs(64.3%) and S-HBs(60.7%) on Day 10. 4) Also, when the in vitro survival of Day 9 HBs was evaluated under different culture condition after thawing, the result in culture medium only (79.3%) was significantly higher than 43.2% in co-culture group (p<0.05). These results demonstrated that bovine HBs can be successfully cryopreserved by two-step vitrification method using EFS35.

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Studies on the Nature and Pathogenicity of Nuclear Polyhedrosis Virus of the Fall Webworm, Hyphantria cunea (Drury) (흰불나방 핵다각체병바이러스의 성상과 병원성에 관한 연구)

  • Im Dae Joon;Hyun Jae Sun;Paik Woon Hah;Lim Jong Sung
    • Korean journal of applied entomology
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    • v.18 no.1 s.38
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    • pp.1-10
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    • 1979
  • An inclusion forming virus isolated from a fan webworm, Hyphantria cunea, in 1975 was identified as a nuclear polyhedrosis virus. With the virus isolated in Korea, it was considered that the virus would be one of the valuable microorganism in microbial control. In this connection, 1) the shape and size of the virus for identification, 2) susceptibility of the various instar larvae to the virus, 3) the effects of storage condition on the pathogenicity and the cross infection of the virus to the larvae of Bombyx mori were examined. The results are summarized as follows; 1. The polyhedron was tetrahedron or hexahedron of $2\mu$ in size and the rod-shaped virus particles consisting of $2\~14$ rods in a bundle were $330m{\mu}\times35m{\mu}$ in size. 2. The hexagonal nuclear polyhedra were found only in the nucleus of the midgut cells but were variable in size. 3. The $LD_{50}$ values for the various instar larvae of H. cunea were $8.377\times10^4\;PIBs/ml$ for the second, $4.974\times10^5\;PIBs/ml$ for the fifth instar larvae. The $LT_{50}values$ for $10^6\;PIBs/ml$ were 9.6 days for the second, 11.5 days for the third, 12.0 days for the fourth and 17 days for the fifth instar larvae. 4. The susceptibility of H. cunea to the nuclear polyhedrosis virus was greater in the first generation than in the second generation. 5. The effect of the storage conditions on the pathogenicity of the nuclear polyhedra was less in refrigerator $(5^{\circ}C)$ and in freezing $(-80^{\circ}C)$ than in room temperature $(18.5^{\circ}C)$, especially as air-dried polyhedra than as suspension. The pathogenicity of the polyhedra seemed to decrease by sunlight during storage as cadavers, since rather greater decrease in pathogenicity was found in sunny condition than in shady condition. 6. The effective spray concentration was $6.4\times10^7\;PIBs/ml$ in the field and its $LT_50$ values for the third and the fifth instar larvae were 4.8 days and 14.2 days, respectively. 7. No cross infections were found in the nuclear polyhedrosis virus between H. cunea and B. mori. larvae.

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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.

Investigation of ground condition charges due to cryogenic conditions in an underground LNG storage plant (지하 LNG 저장 시험장에서 극저온 환경에 의한 지반상태 변화의 규명)

  • Yi Myeong-Jong;Kim Jung-Ho;Park Sam-Gyu;Son Jeong-Sul
    • Geophysics and Geophysical Exploration
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    • v.8 no.1
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    • pp.67-72
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    • 2005
  • To investigate the feasibility of a new concept of storing Liquefied Natural Gas (LNG) in a lined hard rock cavern, and to develop essential technologies for constructing underground LNG storage facilities, a small pilot plant storing liquid nitrogen (LN2) has been constructed at the Korea Institute of Geoscience and Mineral Resources (KIGAM). The LN2 stored in the cavern will subject the host rock around the cavern to very low temperatures, which is expected to cause the development of an ice ring and the change of ground condition around the storage cavern. To investigate and monitor changes in ground conditions at this pilot plant site, geophysical, hydrogeological, and rock mechanical investigations were carried out. In particular, geophysical methods including borehole radar and three-dimensional (3D) resistivity surveys were used to identify and monitor the development of an ice ring, and other possible changes in ground conditions resulting from the very low temperature of LN2 in the storage tank. We acquired 3D resistivity data before and after storing the LN2, and the results were compared. From the 3D images obtained during the three phases of the resistivity monitoring survey, we delineated zones of distinct resistivity changes that are closely related to the storage of LN2. In these results, we observed a decrease in resistivity at the eastern part of the storage cavern. Comparing the hydrogeological data and Joint patterns around the storage cavern, we interpret this change in resistivity to result from changes in the groundwater flow pattern. Freezing of the host rock by the very low temperature of LN2 causes a drastic change in the hydrogeological conditions and groundwater flow patterns in this pilot plant.

A Study on the Characteristics of Ca(OH)2 According to the Calcination Conditions of Oyster Shells and Its Application for Exterior Water Paints (굴 패각의 소성 조건에 따른 소석회의 특성과 외부용 수성 도료 적용 연구)

  • Hwang, Dae Ju;Yu, Young Hwan;Han, Chang Soo;Lee, Jong Dae
    • Korean Chemical Engineering Research
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    • v.60 no.4
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    • pp.594-605
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    • 2022
  • CaO was prepared by calcining for oyster shells using a microwave kiln. It was analyzed to Ca(OH)2 synthed on hydration reaction from prepared CaO. The synthesized Ca(OH)2 was formulated as an external water paint. Oyster shells (325 mesh, 43 ㎛) were decarbonized for (a) 950 ℃/1 hr and (b) 1,150 ℃/1 hr to prepare CaO. In the calcination condition of (a), CaO was 56.7 wt%, and in the calcination condition of (b), CaO was 100 wt%. To compare CaO by calcination of oyster shells with that of limestone, limestone (25~30 mm) was decarbonized at 950 ℃/1 hr to prepare CaO, and as a result of the analysis(XRD), it was analyzed as CaO 100 wt%. CaO was prepared under the calcining conditions of oyster shells (b) 1,150 ℃/1 hr, and Ca(OH)2 was synthesized through hydration. Hydration conditions of the prepared CaO were (a) CaO : H2O(100 g : 200 g) and (b) CaO : H2O(100 g : 400 g). As a result of the hydration reaction, it was confirmed as low reactivity. 100 wt% of Ca(OH)2 was synthesized. In particular, Ca(OH)2 synthesized under the hydration condition of (a) was analyzed in a plate shape. An external water paint was formulated with Ca(OH)2 synthesized from oyster shells as the main component. When 15 items of the external water paint standard specification (KS M 6010) were analyzed, it was confirmed that all other criteria were satisfied except for freezing stability.

A study for Shear Strength Characteristics of Frozen Soils under Various Temperature Conditions and Vertical Confining Pressures (동결온도조건 및 수직구속응력에 따른 동결토의 전단강도 변화에 관한 연구)

  • Lee, Joonyong;Choi, Changho
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.11
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    • pp.51-60
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    • 2012
  • In order to characterize the shear strength of the frozen sand for foundation design in cold region and prediction of adfreeze bond strength, many researchers developed test techniques and carried out many tests to analyze shear strength properties of the frozen sand for half a century. However, many studies for shear strength properties of the frozen sand have been carried out with limited circumstances, even though shear strength of the froze sand can be affected by various influence factors such as soil type, temperature conditions, and magnitude of normal stress. In this study, direct shear test equipment was used to analyze the shear strength characteristics of the frozen sand. Direct shear test equipment was designed for cold weather, and the direct shear tests were carried out inside of large-scaled low temperature chamber. Three soil types-two uniform sands and one well graded soil were used to analyze the shear strength of the frozen sand with three different temperature conditions and three different vertical confining pressures. In this research, a series of direct shear tests for shear strength of the frozen sand have been conducted to demonstrate the efficiency of effectiveness of the test equipment and low temperature chamber. This research also showed that shear strength of the froze sand increased with decreasing temperature condition, but the influence of vertical confining pressure was insignificant to the shear strength of the frozen sand.

Experimental Investigation of Frost Heaving Susceptibility with Soils from Terra Nova Bay in Eastern Antarctica (동남극 테라노바만 흙 시료의 동상특성에 관한 실험적 연구)

  • Hong, Seungseo;Park, Junghee;Lee, Jongsub;Lee, Jangguen;Kang, Jaemo;Kim, Youngseok
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.12
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    • pp.5-16
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    • 2012
  • The second scientific antarctic station of South Korea is under construction at Terra Nova Bay located in eastern Antarctica. Ground condition in the Antarctica is frozen in general, but there are seasonal frozen grounds with active layers sporadically. When the active layer is frozen, frost heaving occurs that might cause the differential movement of frozen ground and the failure of structures. Therefore, it is necessary to determine the frost heaving susceptibility of soils at Terra Nova Bay before starting antarctic station construction. This study presents experimental investigation of the frost heaving susceptibility of soil samples with variation of particle sizes and unfrozen water contents. The soil samples were taken from five different locations at Terra Nova Bay and physical properties, unfrozen water content, and frost heaving tests were performed. For the frost heaving tests, soil specimens were frozen with constant freezing temperatures at the top and with drainage at the bottom in order to stimulate the frost heaving. The frost heaving tests provide volume expansion, volumetric strain, and heaving rate which can be used to analyze the relationship between the frost heaving vs. particle size and the frost heaving vs. unfrozen water content. Experimental results show that the more the fine contents exist in soils, the more frost heaving occurs. In addition, the frost heaving depends on unfrozen water content. Experimental data can be used to evaluate the frost heaving susceptibility of soils at the future construction site in the Antarctica.

Study on simultaneous determination of aromatic material causing allergic in children's products by GC-MSD (GC-MSD를 이용한 어린이 제품 중 알러지 유발 방향성 물질의 동시분석법 연구)

  • Ko, Kyeong Mok;Rhu, Chan Joo;Ko, Byeong Rae;Lee, Seok Ki
    • Analytical Science and Technology
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    • v.33 no.3
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    • pp.134-142
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    • 2020
  • Twenty-one allergy-induced aromatic material in children's products were analyzed using gas chromatography mass spectrometer(GC-MSD). The analytes were extracted using an automatic Soxhlet extractor, centrifuged for 10 minutes in a fast freezing centrifuge, and the supernatant was filtered with a syringe filter and then transferred into a 2 mL vial and injected in a split mode. In the established condition, the calibration curve showed linearity with a determination coefficient of 0.9981 or more. Sensitivity was 0.3145 ~ 1.6757, which showed a fairly wide range of sensitivity for each substance. The detection limit of the device was 0.0016 ~ 0.0423 ㎍/mL and the maximum detection limit was less than 0.05 ㎍/mL. The method detection limit ranged from 0.0030 ~ 0.0589 ㎍/mL. In addition, the limit of quantification ranged from 0.0096 to 0.1876 ㎍/mL, with precision ranging from 0.41 to 10.49 % and accuracy ranging from 83 to 116 %. The analytical method developed in this study was applied to commercial products.

Effect of Freeze Injury by Weather on Quality of 'Fuyu' Sweet Persimmon at Harvest and Subsequent Storage (수확기 동상해 피해가 '부유' 단감의 저장 품질에 미치는 영향)

  • Jo, Yun Hee;Choi, Seong-Tae;Kim, Seong-Cheol;Kim, Eun-Gyeong;Hwang, Yeon-Hyeon;Kim, Eun-Seok;Ahn, Gwang-Hwan
    • KOREAN JOURNAL OF PACKAGING SCIENCE & TECHNOLOGY
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    • v.21 no.2
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    • pp.73-78
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    • 2015
  • Persimmons on the trees are often exposed to weather condition of cold temperature before harvest, which affects the fruit quality at harvest time and later storage. This study investigated the effect of the harvest season temperature on the storage quality of 'Fuyu' sweet persimmons. Monitoring the temperature changes in the orchard showed five times of cold temperature below the freezing temperature of persimmon, $-2.1^{\circ}C$, in the harvesting period between mid October and mid November with the lowest being as low as $-5.4^{\circ}C$. Among three different harvestings, the tertiary one at mid November showed decreased 'L' value and consistently high 'a' value of color index and high reduction in hardness during 150 days storage of fruits at $0^{\circ}C$ showing freeze-injured symptoms, compared to the first one at mid October and the second one at early November. The fruits of first harvest maintained consistently hard texture with increased 'a' color value probably due to postharvest ripening. The tertiary harvest exhibited increase in fructose and glucose contents and decrease in sucrose of the fruits, being very different from the first and secondary harvests which maintained constant level of all three free sugars. Mid November with possible sudden temperature drop is unsuitable period for harvesting sweet persimmons in the quality at harvest and later storage, and thus the harvest should be designed to be done before that time.

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Characterization of Materials for Retort Processing in Oyster Porridge (레토르트 굴죽 제조를 위한 원료의 가공적성)

  • 허성호;이호재;홍정화
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.5
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    • pp.770-774
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    • 2002
  • The effect of mixing ratio of materials and their retort processing conditions on the physical properties and preference of oyster porridge were investigated. Rice gave adequate viscosity (about 800 cp) to the porridge as it was used 10~12% level and half of it was ground. Potato starch, waxy corn starch, and Perfectamyl AC showed small variation in viscosity unrelated to the temperature, that was a good condition for high quality porridge. Especially, waxy corn starch had a good property for retort porridge because its viscosity was maintained high during the processing and decreased to preferable one after processing. Purity CSC, modified starch gave gum-like texture and didn't cause water-separation much after freezing and thawing treatment of the porridge. Therefore, it was good for enhancing the physical property of the porridge as a co-additive of waxy corn starch when it was used at quarter level of waxy corn starch. Xanthan gum increased dispersibility of materials and it was useful for stabilizing physical quality of the porridge without affecting preference when it was used below 0.2% concentration. As a result, the optimal material-mixing ratio was determined as rice 10% (50% of it was ground), waxy corn starch 1.5%, Purity CSC 0.5%, xmthan gum 0.2%, salt 0.3%, and water 87.5%. Oyster's porridge, retort processed with this recipe, showed stable physical property after 6 month storage at room temperature.