• Title/Summary/Keyword: Thawing method

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Production and Cryopreservation of Sexed Embryos after Micromanipulative Biopsy and PCR (미세조작 및 PCR 기법을 이용한 성판별 수정란의 생산 및 동결)

  • 이홍준;서승운;김기동;이상호
    • Journal of Embryo Transfer
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    • v.15 no.2
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    • pp.175-180
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    • 2000
  • The possible use of micromanipulative biopsy and PCR of the biopsied embryonic cells was tested to produce sexed bovine embryos in practical terms. By micromanipulation and PCR techniques, higher survival rate and accurate sexing of demi-embryos were btained. Bovine oocytes matured and fertilized in vitro were co-cultured with bovine oviductal epithelial cell (BOEC) monolayer in USU-6 medium supplemented with 15% FBS, and the embryos of 37% (327/885) were developed to blastocysts. Among 111 blastocysts produced by invitro, only 7 (6.3%) embryos were found unable to determine their sex, probably due to the loss of cells, since no PCR product was found from those cells. All the remaining 104 (93.7%) demi-embryos survived micromanipulation and demonstrated male-specific product or bovine-specific product alone suggesting that correct sexing of the sample. Forty-three point one percent(25/58) of manipulated and cryopreserved demi-embryos after thawing were survived. Final verification of the sexed embryos is necessary to make sure the same sex in fetus and newborn calf upon embryo transfer. The established sexing method on a large number of bovine embryos from previous and this study suggests that this a could be used practically in the field.

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Immune Status of Breeding Hens Against Newcastle Disease (종계군(種鷄群)의 Newcastle Disease에 대(對)한 면역상태(免疫狀態)에 관(關)한 연구(硏究))

  • Yeo, Sang Geon;Choi, Won Pil
    • Korean Journal of Veterinary Research
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    • v.19 no.1
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    • pp.45-51
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    • 1979
  • The present study was undertaken to observe the immune status of breeding hens and laying hens against Newcastle disease (ND). The methods of extraction of hemagglutination inhibition (HI) antibody from egg yolk, the detection of HI antibody in egg albumen and the correlation between HI antibody titers in maternal sera and egg yolks were discussed. For the purposes of these experiments, 9 flocks of breeding hens and 16 flocks of laying hens immunized against Newcastle disease virus were investigated. The vaccination program of tested flocks was 3-3-3 or 4-4-4 in general. The results obtained are summerized as follows: Freezing-thawing was the best method far antibody extraction from egg yolk for HI test. The HI antibody against NDV was found in egg albumen (geometric mean, 4.5), but lower than that found in egg yolk (32.1). The geometric mean of HI antibody titers of egg yolks (84.1) was higher than that of maternal sera (68.4) and day-old chicken sera (25.3). There was correlation between HI antibody titers of maternal sera(Y) and those of egg yolks(X). The coefficient correlation was r=0.63, and the line of regression of Y on X was $\hat{Y}$=35.91+0.35X.

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Immunoliposomes Carrying Plasmid DNA : Preparation and Characterization

  • Kim, Na-Hyung;Park, Hyo-Min;Chung, Soo-Yeon;Go, Eun-Jung;Lee , Hwa-Jeong
    • Archives of Pharmacal Research
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    • v.27 no.12
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    • pp.1263-1269
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    • 2004
  • The objective of this study was to characterize immunoliposomes carrying plasmid DNA with optimal encapsulation efficiency and antibody density. Plasmid DNA was encapsulated by the freezing/thawing method into liposomes composed of POPC (1-palmitoyl-2-oleoyl-sn-glycerol- 3-phosphocholine), DDAB (didodecyl dimethyl ammonium bromide), DSPE-PEG 2000 (distearoyl phosphatidyl ethanolamine polyethylene glycol 2000) and DSPE-PEG 2000-maleimide. The liposomes carrying plasmid DNA were extruded through two stacked polycarbonate filters, of different pore size, to control the liposome size. Then, rat IgG molecules were conjugated to the liposomes. The immunoliposomes containing plasmid DNA were separated from the free plasmid DNA and unconjugated IgG by Sepharose CL-4B column chromatography. The DNA amount encapsulated was affected by DDAB (cationic lipid) concentration, the initial amount of plasmid DNA between 10 ${\mu}g$ and 200 ${\mu}g$, the total lipid amount and plasmid DNA size, but not significantly by liposome size. By varying the ratio of DSPE-PEG 2000-maleimide to IgG, the number of IgG molecules per liposome was changed significantly.

Effects of Artificial Supercooling Followed by Slow Freezing on the Microstructure and Qualities of Pork Loin

  • Kim, Yiseul;Hong, Geun-Pyo
    • Food Science of Animal Resources
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    • v.36 no.5
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    • pp.650-655
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    • 2016
  • This study investigated the effects of artificial supercooling followed by still air freezing (SSF) on the qualities of pork loin. The qualities of pork frozen by SSF were compared with the fresh control (CT, stored at 4℃ for 24 h), slow freezing (SAF, still air freezing) and rapid freezing (EIF, ethanol immersion freezing) treatments. Compared with no supercooling phenomena of SAF and EIF, the extent of supercooling obtained by SSF treatment was 1.4℃. Despite that SSF was conducted with the same method with SAF, application of artificial supercooling accelerated the phase transition (traverse from -0.6℃ to -5℃) from 3.07 h (SAF) to 2.23 h (SSF). The observation of a microstructure indicated that the SSF prevented tissue damage caused by ice crystallization and maintained the structural integrity. The estimated quality parameters reflected that SSF exhibited superior meat quality compared with slow freezing (SAF). SSF showed better water-holding capacity (lower thawing loss, cooking loss and expressible moisture) and tenderness than SAF, and these quality parameters of SSF were not significantly different with ultra-fast freezing treatment (EIF). Consequently, the results demonstrated that the generation of supercooling followed by conventional freezing potentially had the advantage of minimizing the quality deterioration caused by the slow freezing of meat.

Optimization of Preparation Variables for Trimyristin Solid Lipid Nanoparticles

  • Choi, Mi-Hee;Lee, Mi-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.37 no.1
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    • pp.51-55
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    • 2007
  • Solid lipid nanoparticles (SLNs) have been regarded to behave similar to the vegetable oil emulsions because emulsions of lipid melts are formed before lipid droplets being solidified to turn into SLNs. Compared to lipid emulsion, however, it has been more difficult to obtain stable SLNs and needs more extensive considerations on stabilizer and manufacturing process. In the present study, we tried to prepare phosphatidylcholine-based trymyristin (TM) SLNs using high pressure homogenization method and optimize the manufacturing variables such as homogenization pressure, number of homogenization cycles, cooling temperature, co-stabilizer and freeze-drying with cryoprotectants. Nano-sized TM particles could be Prepared using egg Phosphatidylcholine and pegylated phospholipids ($PEG_{2000}$PE) as stabilizers. Based on the optimization study, the dispersion was manufactured by homogenization under the pressure of 100 MPa for more than 5 cycles, and solidifying the intermediately formed lipid melt droplets by dipping in liquid nitrogen followed by thawing at room temperature. In addition, TM SLNs could be freeze-dried and then redispersed easily without significant particle size changes after freeze drying with 10% and 12.5% sucrose or trehalose. The TM SLNs established in this study can be used as delivery system for drugs and cosmetics.

Analysis of Frozen Reduction Effect and Economic Evaluation of Recycled PET-Soil (재활용 PET 재료를 이용한 골재의 동상저감 효과 분석 및 경제성 평가)

  • Shin, Eun Chul;Shin, Hui Su;Kim, Gi Sung
    • Journal of the Korean Geosynthetics Society
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    • v.13 no.4
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    • pp.153-159
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    • 2014
  • During the winter and spring seasons in Korea, structures such as buried water supply pipelines, roads, railways are frequently damaged due to frost heaving and thawing. Until now, the method of substituting the frost susceptible soil with the gravel or rubbles those are non-frost susceptible materials have been employed in Korea to prevent frost heaving. A series of laboratory soil tests and indoor frozen soil engineering experiments, as well as laboratory frost heaving tests were conducted for seeking the means of utilizing recycled PET bottles as substitute material.

Comparison of Spermatozoa Recovery Methods on Cauda Epididymal Sperm of Hanwoo Bulls

  • Kang, Sung-Sik;Kim, Ui-Hyung;Jeon, Min-Hyeong;Lee, Myung-Suk;Cho, Sang-Rae
    • Journal of Embryo Transfer
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    • v.33 no.4
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    • pp.321-326
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    • 2018
  • In this study, two epididymal spermatozoa recovery methods in relation to sperm number, motility, viability and acrosome reaction were examined. Seven bulls were castrated and 7 testicles with epididymides were transferred to the laboratoy. Epididymis in each bull was randomly used for flushing and mincing methods with semen extender (Optixcell, IMV, France). The recovered spermatozoa with adjusted sperm concentration to $40{\times}10^6cells/mL$ was diluted with optixcell and cryopreserved. In experiment 1, the difference in the total number of spermatozoa using flushing and mincing methods was insignificant (2570.0 and $2505.2{\times}10^6cells/mL$, respectively). For experiment 2, the percentage of motile spermatozoa and motility parameters between flushing and mincing methods were studied through the use of sperm class analyzer after frozen-thawing. The percentage of total motile sperm between flushing and mincing methods was almost the same with $89.5{\pm}12.8$ and $91.4{\pm}7.9%$, respectively. The same is the case with experiment 3 wherein the viability and acrosomal integrity of frozen-thawed epididymal spermatozoa by flushing and mincing was insignificantly different. The results from the study showed that both flushing and mincing methods can be used for epididymal spermatozoa recovery in bull.

Landslide Prediction with Angle of Repose Prediction Using 3D Spatial Coordinate System and Drone Image Detection (3차원 공간 좌표 시스템과 드론 영상 검출을 활용한 산사태 안식각 예측에 관한 연구)

  • Yong-Ju Chu;Soo-Young Lim;Seung-Yop Lee
    • Smart Media Journal
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    • v.12 no.3
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    • pp.77-84
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    • 2023
  • Forest fires are representative natural disasters resulting from dramatic global climate change in these modern times. When forest formation is insufficient due to forest damage caused by fire, secondary damages such as landslides occur during the winter thawing period and heavy rains. In most countries, only a limited area is managed as CCTV-centered monitoring systems for forest management. For the landslide prediction, markers containing 3D spatial coordinates were located on the slopes of the danger areas in advance. Then 3D mapping and angle of repose were obtained by periodic drone imaging. The recognition range and angle of view of markers were defined, and a new method for predicting signs of landslides in advance was presented in this study.

Clinical Application of Oocyte Cryopreservation I. Pregnancy and Delivery of Vitrified Human Oocytes in ART Program (난자동결보존의 임상적 응용 I. 유리화 난자동결 보존에 의한 임신과 분만)

  • 정형민;박이석;차광렬
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.245-250
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    • 2001
  • This study was performed to evaluate whether vitrification method using ethyle glycol and eletron microscopic (EM) grid could be used far the cryopreservation of human oocytes in ART program. Surplus oocytes were obtained from consented IVF patients. These surplus human oocytes were frozen with our vitrification method, Oocytes were exposed to 1.5M ethylene glycol (EG) in DPBS far 2,5 minutes, followed by 5.5M EG plus 1.0M Sucrose in DPBS for 20 seconds. Then oocytes were transferred onto the EM grid and the grid was plunged into LN2 for storage. For thawing, oocytes containing EM grid were sequentially transferred in 1.0M, 0.5M, 0.25M, 0.125M and 0 M sucrose in DPBS solution at the intervals of 2.5 minutes. Thawed and survived oocytes were provided for ICSI. Embryos from vitrified oocytes were transferred to uterus of the patient on 4 to 5 days after ovulation in natural cycles of on 15 to 17 day of hormone replacement cycles. A total of 370 oocytes from 26 patients were thawed and 159 (43.0%) of them survived. One hundred thirty four oocytes (84.3%) were fertilized normally and 126 pre-embryos were transferred to 26 patients, resulting in 5 clinical pregnancies. The pregnancy rate per transfer was 19.2% and implantation rate was 4.0%. Among the five pregnant, 4 patients delivered 4 healthy babies and the one patient was 32-week ongoing pregnancy. From this results, vitrification using ethylene glycol as cryoprotectant and EM grid is a rapid and simple method that can be effectively applied for the cryopreservation of human oocytes in ART program.

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Bioavailability of Tripotassium Dicitrato Bismuthate by ICP-MS in Human Volunteers (ICP-MS를 사용한 구연산비스마스칼륨 (Tripotassium dicitrato bismuthate)의 생체이용률 측정)

  • Kwon, Oh-Seung;Kwon, Jee-Young;Yoon, Ae-Rin;Park, Kyung-Soo
    • Journal of Pharmaceutical Investigation
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    • v.37 no.2
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    • pp.79-84
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    • 2007
  • This study was aimed to establish analytical method of Bi to develop a guideline of the bioequivalence test of tripotassium dicitrato bismuthate (TDB). For this purpose, a simple, specific and sensitive inductively coupled plasma-mass spectrometry (ICP/MS) method were developed and validated in human plasma. Various concentrations of bismuth standard solution (0-25ng/mL) were prepared with distilled water and human blank plasma. To 10mL of the volumetric flasks, 2mL of blank plasma was added with 8ml of distilled water. Bi standard solution was added to prepare the calibration samples and injected into ICP-MS. The plasma samples obtained from volunteers given 3 tablets of bismuth (total 900mg as TDB) were analyzed as described above. As a result, the coefficients of variation were <20% in quantitation limit (0.2 ng/mL) and <15% at the rest of concentrations. The stability test by repeated freezing-thawing cycles showed that the samples were stable only for 24hr. The stability tested for samples with a short-term period of storage at room temperature and pre-treatment prior to the analysis showed very stable over 24hr. In 8 healthy Korean subjects received Denol tablets at the dose of 900mg bismuth, AUC, $C_{max},\;T_{max}$ and half-life $(t_{1/2})$ were determined to be $198.33{\pm}173.78 ng{\cdot}hr/mL,\;64.48{\pm}27.06 ng/mL,\;0.52{\pm}0.21 hr,\;and\;5.15{\pm}2.67 hr$, respectively, from the plasma bismuth concentration-time curves. In conclusion, the method was suitable for the determination of bismuth in human plasma samples and could be applied to bioequivalence test of bismuth tablet.