• Title/Summary/Keyword: Frozen solution

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Studies on the Properties of Simmental Semen I. With Special Reference to Collecting Semen During Summer (Simmental의 정액성상에 관한 연구 제1보, 정액의 하계수취를 중심으로)

  • 고광두;한두희;정길생
    • Korean Journal of Animal Reproduction
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    • v.5 no.2
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    • pp.43-48
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    • 1981
  • This experiment was carried out with the purpose to get some information about the properties of Simmental bull semen during summer season. The results obtained were summarized as follows: 1. Semen volume per ejaculation, sperm concentration and sperm viability were averaged 5.16ml, 6.6billion and 65%, respectively. 2. Percentage of motile sperm after dilution in skmmilk solution and trisbuffer for 5 days were 34.16% and 35.0%, respectively. 3. Viability of spermatozoa frozen in skimmilk extender and trisbuffer for 5 days were 20. 83% and 25.66%, respectively. 4. Percentage of live sperm, MRT and pH value were 71.8∼72.1%, 8.40∼8.21 minutes and 6.78, respectively. 5. Diluted semen showed strong resistance to the cold shock than that of fresh semen. 6. Rscovery of sperm motility after freezing for 24 hours was relatively weak.

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Solid State Dynamic Nuclear Polarization of 1H Nuclear Spins at 0.3 T and 4.2 K

  • Shim, Jeong Hyun
    • Journal of the Korean Magnetic Resonance Society
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    • v.21 no.4
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    • pp.114-118
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    • 2017
  • Here, I report solid state Dynamic Nuclear Polarization (DNP) of $^1H$ nuclear spins at 0.3 T and 4.2 K. The DNP polarizer was developed based on a commercial X-band Electron Spin Resonance (ESR) modified for DNP, in combination with a NMR console and a liquid-Helium cryostat. By detuning magnetic field, DNP spectrum was measured to find the optimal condition. At +3 mT detuned from on-resonance field, $^1H$ NMR signal of 60:40 glycerol/water frozen solution doped with 20 mM perdeuterated-Tempone was amplified 43 times. The $^1H$ spin polarization obtained at 4.2 K is over 3100 times higher than that at 300 K. The width of the DNP spectrum, which is five times broader than ESR spectrum, is inconsistent with solid effect or thermal mixing, and presumably suggests a different DNP mechanism.

RBF Network Based QFT Parameter-Scheduling Control Design for Linear Time-Varying Systems and Its Application to a Missile Control System (시변시스템을 위한 RBF 신경망 기반의 QFT 파라미터계획 제어기법과 alt일 제어시스템에의 적용)

  • 임기홍;최재원
    • 제어로봇시스템학회:학술대회논문집
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    • 2000.10a
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    • pp.199-199
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    • 2000
  • Most of linear time-varying(LTV) systems except special cases have no general solution for the dynamic equations. Thus, it is difficult to design time-varying controllers in analytic ways, and other control design approaches such as robust control have been applied to control design for uncertain LTI systems which are the approximation of LTV systems have been generally used instead. A robust control method such as quantitative feedback theory(QFT) has an advantage of guaranteeing the stability and the performance specification against plant parameter uncertainties in frozen time sense. However, if these methods are applied to the approximated linear time-invariant(LTI) plants which have large uncertainty, the designed control will be constructed in complicated forms and usually not suitable for fast dynamic performance. In this paper, as a method to enhance the fast dynamic performance, the approximated uncertainty of time-varying parameters are reduced by the proposed QFT parameter-scheduling control design based on radial basis function (RBF) networks for LTV systems with bounded time-varying parameters.

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Viability of Mouse Morula Embryos Frozen Rapidly in Liquid Nitrogen Vapour (급속동결한 마우스 상실배의 체외배양후 생존성에 관하여)

  • 신상태
    • Journal of Embryo Transfer
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    • v.4 no.1
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    • pp.35-40
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    • 1989
  • The effects of cryoprotectants (glycerol, DMSO and ethylene glycol) and the concentrations (0, 0, 25, 0.5and 1.0 M) of sucrose in the diluent on the is vitro survival of mouse morulae froaen rapidly in liquid nitrogenvapour were examined. When the embryos were equilibrated in 1.5 M cryoprotectants +0.25 M sucrose in one-step or in 3.0 M cryoprotectants +0.25 sucrose in two-step and diluted with 0, 0.25, 0.5, or 1.0 M sucrose solution after thawing, high survival rates were obtained in ethylene glycol (48.0% to 88.2 %) or in glycerol (35.0 % to 77.8 %). These results show that 1.5 M ethylene glycol is a highly efficient cryoprotective agent for the rapid freezing of mouse morula embryos and 0.5 M sucrose was optimal concentration in the diluent after thawing.

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Effect on the Adhesion of Ice Slurry by the Characteristic of Cooling Surface (냉각면 성상이 빙부착에 미치는 영향)

  • Seung Hyun;Hong Hi Ki;Kang Chae Dong
    • Korean Journal of Air-Conditioning and Refrigeration Engineering
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    • v.17 no.2
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    • pp.183-190
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    • 2005
  • In the process of ice-slurry making, ice adhesion on cooling wall or in narrow flow Path disturbs continuous ice formation. In this study, the effect on the ice adhesion to cooling surface by some freezing experiments was investigated, quantitatively. Three types of solutions were frozen in various coating vessels with stirring. In the experiment, the ice adhesion between cooling wall and Ice-slurry was evaluated by measuring stirring power. From the result, the stirring power of slurry mixture in PTFE-coating vessel was smaller than those in PE-coating, PFA-coating and bare SUS vessel. Especially, in EG H PG 1.S/ HD 1.5 mass$\%$ solution, the stirring power in the PE-coating vessel was smaller than that in the PFA-coating or SUS vessel.

Numerical Analysis of Hypersonic Flow over Small Radius Blunt Bodies (작은 크기의 무딘 물체에 대한 극초음속 유동의 수치해석)

  • Lee Chang Ho;Park Seung O
    • 한국전산유체공학회:학술대회논문집
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    • 2002.05a
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    • pp.109-114
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    • 2002
  • The effect of nose radius on aerodynamic heating are investigated by using the Wavier-Stokes code extended to thermochemical nonequilibrium airflow. A spherical blunt body, whose radius varies from 0.003048 m to 0.6096 m, flying at Mach 25 at an altitude of 53.34 km is considered. Comparison of heat flux at stagnation point with the solution of Viscous Shock Layer and Fay-Riddell are made. Obtained result reveals that the flow chemistry for very small radius is nearly frozen, and therefore the contribution of heat flux due to chemical diffusion is smaller than that of translational energy. As the radius becomes larger, the portion of diffusion heat flux becomes greater than translational heat flux and approaches to a constant value.

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Analysis of the Gene Expression by Laser Captured Microdissection (I): Minimum Conditions Required for the RNA Extraction from Oocytes and Amplification for RT-PCR (Laser Captured Microdissection을 이용한 유전자 발현에 대한 연구 (I): RT-PCR을 위한 난자의 RNA 추출 및 증폭을 위한 최소한도의 확립)

  • Park, Chang-Eun;Ko, Jung-Jae;Cha, Kwang-Yul;Lee, Kyung-Ah
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.3
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    • pp.183-190
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    • 2001
  • Objective: Recently, microdissection of tissue sections has been used increasingly for the isolation of morphologically identified homogeneous cell populations, thus overcoming the obstacle of tissue complexity for the analysis cell-specific expression of macromolecules. The aim of the present study was to establish the minimal conditions required for the RNA extraction and amplification from the cells captured by the laser captured microdissection. Methods : Mouse ovaries were fixed and cut into serial sections (7 im thickness). Oocytes were captured by laser captured microdissection (LCM) method by using PixCell $II^{TM}$ system. The frozen sections were fixed in 70% ethanol and stained with hematoxylin and eosin, while the paraffin sections were stained with Multiple stain. Sections were dehydrated in graded alcohols followed by xylene and air-dried for 20 min prior to LCM. All reactions were performed in ribonuclease free solutions to prevent RNA degradation. After LCM, total RNA extraction from the captured oocytes was performed using the guanidinium isothiocyanate (GITC) solution, and subsequently evaluated by reverse transcriptase-polymerase chain reaction (RT-PCR) for glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). Results: With the frozen sections, detection of the GAPDH mRNA expression in the number of captured 25 oocytes were not repeatable, but the expression was always detectable from 50 oocytes. With 25 oocytes, at least 27 PCR cycles were required, whereas with 50 oocytes, 21 cycles were enough to detect GA PDH expression. Amount of the primary cDNA required for RT-PCR was reduced down to at least 0.25 $\grave{i}$ l with 50 oocytes, thus the resting 19.75 il cDNA can be used for the testing other interested gene expression. Tissue-to-slide, tissue-to-tissue forces were very high in the paraffin sections, thus the greater number of cell procurement was required than the frozen sections. Conclusion: We have described a method for analyzing gene expression at the RNA level with the homogeneously microdissected cells from the small amount of tissues with complexity. We found that LCM coupled with RT-PCR could detect housekeeping gene expression in 50 oocytes captured. This technique can be easily applied for the study of gene expression with the small amount of tissues.

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In vitro Fertilization and Embryo Development in Simple Media of the Frozen-Thawed Cumulus-free Mouse Oocytes Cryopreserved by Vitrification (Cumulus Free 생쥐 성숙란의 초자화 동결-융해 후 Simple Media에서의 수정 및 배 발달)

  • Jung, Soo-Kyung;Kim, Sung-Kun;Lee, Jung-Jae;Oh, Ji-Hyun;Lee, Yong-Ho;Kim, Sun-Haeng
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.3
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    • pp.201-207
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    • 2002
  • Objective: To observe the capability of fertilization and embryo development including blastocyst formation of the oocytes in simple media after thawing of the cryopreserved cumulus-free mouse oocytes by vitrification method. Methods: Oocytes were collected from 5 to 6 weeks old ICR female mice, and were denuded from the cumulus cells by 0.1% hyaluronidase. Recovered mature oocytes in study group were cryopreserved by vitrification method using EM grid for $5{\sim}7$ days. In brief, oocytes were exposed in dPBS containing 1.5 M EG and 5.5 M EG+1 M sucrose for 2.5 minutes and 20 seconds each, and then executed vitrification by plunging in LN2 after loading on EM grid. Thawing treated by exposure of 1, 0.5, 0.25 and 0.125 M sucrose solution for 2.5 minutes each in order and used for experiments. Spermatozoa aspirated form the epididymis of 12 weeks old ICR male mice were used for insemination after capacitation. T6 media containing 0.4% BSA were used for fertilization and development. Results: Survival and fertilization rates after thawing were 76.9% and 79.6% respectively. Fertilization rate was lower (p<0.005) than that of control group (92.9%). There was no difference in embryo developmental rates from 2-cell to morula, however, the blastocyst formation rate and mean cell numbers of blastocysts in study group (63.3%, $58.9{\pm}9.2$) were lower compared with those of control group (76.1%, $63.5{\pm}8.9$). Conclusion: Vitrification is an effective method for mouse mature oocyte cryopreservation with high survival and fertilization rate after thawing. And in simple media, fertilization rates and embryo development of frozen-thawed mouse oocytes are satisfactory.

Study on the Quality Improvement of Sardine Surimi (정어리 냉동고기풀의 품질개선에 관한 연구)

  • Oh, Kwang-Soo;Moon, Soo-Kyung;Lee, Eung-Ho;Kim, Bok-Gyu
    • Korean Journal of Food Science and Technology
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    • v.25 no.4
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    • pp.327-333
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    • 1993
  • The processing conditions and quality of sardine surimi were examined: Raw sardine meat was separated, washed in 0.2% $NaHCO_3$ and 0.15% NaCl solution, and then dewatered by centrifuge. The dewatered sardine meat was chopped, mixed with 20% emulsion curd (soybean protein : water : refined sardine oil=1:5:2.6), 4% sorbitol, 4% sucrose, 0.2% polyphosphate and 0.1% sodium erythorbate by stone mortar. The mixed sardine meat was frozen with contact freezer, packed in carton box and then stored at $-25{\pm}2^{\circ}C$. The moisture, crude protein and lipid contents of the sardine surimi product was 73.3%, 15.0% and 6.9%, respectively. Fatty acid composition of product consisted of 28.8% of saturates, 24.3% of monoenes and 47.7% of polyenes and the major fatty acids were 16:0, 20:5, 18:1, 22:6 and 16:1. The results of changes in POV, TBA value, fatty acids, texture and sensory score of products during frozen storage showed that lipid oxidation and freeze denaturation of product could be retarded, and flavor enhanced by addition 20% emulsion curd and 0.1% sodium erythorbate. In an attempt to apply sardine surimi in producing surimi-based product, it was concluded that pollack surimi could be substituted with sardine surimi up to 40% without showing any significant changes in texture and taste of surimi-based product.

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Effect of Hypo-osmotic Swelling (HOS) Test on Subsequent Post-thaw Testicular Spermatozoa (고환조직 동결-융해 후 회수된 고환 정자에 대한 Hypo-osmotic Swelling (HOS) Test의 효과)

  • Park, Yong-Seog;Lee, Hyoung-Song;Song, Sang-Jin;Kim, Jeong-Wook;Kang, Inn-Soo;Seo, Ju-Tae
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.3
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    • pp.267-273
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    • 2000
  • Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.

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