• Title/Summary/Keyword: TBM

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Assessment of the Fertilizing Capacity of Domestic Animal Spermatozoa by Hamster Test II. Effects of incubation medium and X-ray irradiation on hamster test for boar spermatozoa (Hamster test를 이용한 가축정자의 수정능력 검정 II. 정액배지 및 X-선조사가 돼지정자의 Hamster test에 미치는 영향)

  • Kim Yong-Jun;Ji Dong-Boum
    • Journal of Veterinary Clinics
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    • v.9 no.2
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    • pp.373-390
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    • 1992
  • To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.

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Infiltration behavior and face stability of carbonate-added slurry shield tunnel (탄산을 첨가한 슬러리 쉴드 터널에서의 침투 거동 및 굴진면 안정성 평가)

  • Lee, Ik-Bum;Choi, Ki-Hoon;Lee, In-Mo
    • Journal of Korean Tunnelling and Underground Space Association
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    • v.15 no.4
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    • pp.401-413
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    • 2013
  • Slurry shield tunnelling ensures stability by pressurizing the tunnel face with the slurry contained in the chamber. It resists water and earth pressure in order to prevent the failure in the tunnel face during tunnel excavation. If the ground is relatively coarse, slurry can not clog the tunnel face and excessive slurry infiltration will occur. In this case chemical compounds or additives should be added to the slurry in order to improve the clogging phenomena at the tunnel face. In this study, the effect of the carbon dioxide gas as an additive to the slurry instead of chemical compounds on the capability of enhancing the clogging in the tunnel face is investigated. Bubbles arising from the carbonate-added slurry are trapped in the soil voids enhancing the clogging capability. This effect is studied in this paper by performing laboratory model tests simulating in-situ conditions, and by adopting the fine particle clogging theory. Tunnel face stability analysis was also performed and it was found that the effective size ($D_{10}$) of soils which can guarantee tunnel stability utilizing the carbonate-added slurry increased from 1.0 mm up to 2.6 mm. Moreover, Stability analysis showed that the tunnel face is stable if the ${\lambda}$(deposition coefficient) value is greater than $0.007sec^{-1}$.

Comparison of Semen Characteristics, Frozen-Thawed Sperm Viability, Testosterone Concentration and Embryo Development between Yorkshire Boar A and B

  • Yi, Y.J.;Lee, S.H.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.612-616
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    • 2004
  • This study was carried out to compare the semen characteristics, frozen-thawed sperm viability and testosterone concentration and in vitro fertilization (IVF) and development of in vitro matured pig oocytes between two Yorkshire boars. Semen and blood samples were collected once per week from October to November 2002 from two adult Yorkshire boars at 18 months of age with 170 kg body weight. Sperm were deep frozen in 5 ml maxi-straws with lactose-egg yolk and N-acetyl-D-glucosamine (LEN) diluent and stored in liquid nitrogen. Blood samples were obtained at 10 a.m. by inserting a 21 gauge, hypodermic needle attached to 10 ml syringe into surface veins in the ear. The concentration of testosterone was determined by Competitive Enzyme Immunoassay. Ovaries were collected from prepubertal gilts at a local slaughter house. Cumulus oocyte complexes were aspirated from antral follicles (3 to 6 mm in diameter). The medium used for oocyte maturation was modified TCM 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at $38.5^{\circ}C$, 5% $CO_2$ in air. For IVF, one frozen 5 ml straw was thawed at $52^{\circ}C$in 40 sec and was diluted with 20 ml Beltsville thawing solution at room temperature. Sperm were washed 2 times in mTLP-PVA and inseminated without preincubation after thawing. Oocytes were inseminated with $2{\times}10^7$/ml sperm concentration. Oocytes were coincubated for 6 h in 500 ${\mu}$l mTBM fertilization medium. At 6 h after IVF, oocytes were transferred into 500 ${\mu}$l NCSU-23 culture medium for further culture of 48 and 144 h. There were no significant differences in the semen volume, motility, normal acrosome morphology and sperm concentration of raw semen between A and B of Yorkshire boar. However, motility and normal acrosome of boar A were higher than those of boar B at 0.5, 2, 3, 4, 5 and 6 h incubations of frozen-thawed sperm. Testosterone concentration (3.75 ng/ml) of boar A was higher than that (2.34 ng/ml) of boar B. The rate of blastocyst formation (15.1%) of boar A was higher than that (10.4%) of boar B. In conclusion, serum testosterone concentration of boar showed very important role for the frozen-thawed sperm viability and the blastocyst formation of pig oocytes matured in vitro.

Shear Behavior of Sands Depending on Shear Box Type in Direct Shear Test (직접전단실험시 전단상자의 종류에 따른 모래시료의 전단거동)

  • Hong, Young-Ho;Byun, Yong-Hoon;Chae, Jong-Gil;Lee, Jong-Sub
    • Journal of the Korean Geotechnical Society
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    • v.31 no.3
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    • pp.51-62
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    • 2015
  • Shear behavior obtained by direct shear tests is dependent on shear box and boundary condition. The objective of this study is to analyze problems of conventional direct shear test (type-A) and provide the reliable results by developing type-C direct shear apparatus. Experimental tests are carried out for Ulleung sand by using type-A and -C direct shear devices. The soil specimens, which are prepared at the relative density of 60%, and are applied to vertical confining stresses of 50, 100, 200, 300, and 400 kPa, are sheared at a constant shear strain rate of 0.5 mm/min. By comparing the results obtained by type-A and -C direct shear apparatus under constant normal load (CNL) condition, the performance of new one is verified. In addition, two constrained conditions including constant normal load (CNL) and constant pressure (CP) are applied to type-C one. Experimental results show that type-A direct shear apparatus has some problems such as rotating of loading plate and upper shear box, and the frictional forces between soil and inner wall of upper shear box. Thus, the shear strengths obtained by type-A device are overestimated or underestimated depending on shear box and boundary condition. On the other hand, type-C device produces clear and consistent test results regardless of constrained conditions. This study represents that type-C direct shear apparatus not only can solve the problems of type-A direct shear apparatus but provide the reliable results.

Amino Acids Supplemented with Culture Medium Stimulated On Development of Porcine Embryos

  • Lee, Y.S.;S.H. Song;Lee, S.N.;K.H. Chung;Park, C.S.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2002.11a
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    • pp.80-80
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    • 2002
  • This study was carried out that to investigate the effects of amino acids supplemented with culture medium on development of porcine embryos cultured in vitro. Cumulus oocyte complexes (COCs) were cultured in the maturation medium containing hormones (0.5$\mu\textrm{g}$/$m\ell$ LH, 0.5$\mu\textrm{g}$/$m\ell$ FSH and 1$\mu\textrm{g}$/$m\ell$ estradiol-17${\beta}$) for 20-22 h at 39$^{\circ}C$ in an atmosphere of 5% CO$_2$in air. Subsequently, COCs were cultured in hormone-free maturation medium for 20-22 h. After maturation for 40-44h, oocytes were removed cumulus cells by pipetting and cultured with epididymal sperm for 5 h in the mTBM. Embryos obtained were divided in 4 groups (1) cultured in NCSU 23 containing 0.4% BSA to blastocyst stage(Control), (2) essential amino acids (EA), (3) non-essential amino acids (NA), (4) mixture of essential and non essential amino acid (EA+NA). All treated groups(2-4) were used a glucose free NCSU 23 medium supplemented with pyruvate (0.33 mM), lactate (4.5 mM) to morula stage. From morula to blastocyst stage embryos of all treated groups were cultured in NCSU 23 containing 0.4% BSA. The rates of cleaved oocytes at 48 h after IVF were from 82% to 88% in the groups of control, EA, NA and EA+NA, respectively. The in vitro developmental rates into blastocysts in the groups of EA and EA+NA were significantly (P<0.05) higher than those of group of control (35.1, 35.4 vs. 19.4%, respectively), however, no significant (P<0.05) between control and NA. In conclusion, supplemented with essential amino acid or mixture of essential and non essential amino acid in the culture medium at morula stage increased the rate of development to blastocyst on in vitro produced porcine embryos.

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In-Vitro Fertilization and Culture of Pig Oocytes Matured In-Vitro by Liquid Boar Sperm Stored at 4$^{\circ}C$

  • Kim, M. Y.;Y. J. Yi;Y. J. Chang;Park, C. S.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.63-63
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    • 2003
  • This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

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Comparison of the Properties of Youngia sonchifolia Max. For Kimchi Preparation (고들빼기의 김치 가공적성에 관한 연구)

  • 신수철
    • Korean Journal of Plant Resources
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    • v.9 no.2
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    • pp.105-112
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    • 1996
  • Physicochemical properties of Youngia sonchifolia were compared according to harvesting time and brine methods for its Kimchi preparation. Moisture contents in the roots and the leaves of Y. sonchifolia grown for 5 weeks was more than those grown for 11 weeks. 5-week-grown Y. sonchifolia contained more total free sugar than 11-week-grown ones, and among total free sugar the content of fructose was the most, and the contents of glucose and sucrose were rather high. The roots of 11-week-grown Youngia sonchifolia were harder than those of 5-week grown ones. When the roots were soaked in NaCl solution, the hardness decreased with time. Salt concentration of the Kimchi of Y. sonchifolia increased by the brined material of the Kimchi with soaking time.Salt concentration of the Kimchi was higher in the leaves than in the roots.The roots and leaves of Y.sonchifolia grown for 11 weeks contained organic acids such as malic acid, oxalic acid, succinic acid,lactic acid, and citric acid,among which major organic acids were malic acid, oxalic acid, succinic acid, whereas lactic acid and citric acid were minor organic acids.

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Effects of Some Factors on In Vitro Production of Embryos from Antral Follicle-Derived Porcine Oocytes I. Effects of Maturation Media on In Vitro Maturation, Fertilization and Development (돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향 I. 체외성숙, 체외수정, 체외발달에 대한 체외성숙 배양액의 영향)

  • Yeon, S.-H.;Choi, S.-H.;Kim, C.-D.;Son, D.-S.;Han, M.-H.;Lee, K.-S.
    • Journal of Embryo Transfer
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    • v.19 no.2
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    • pp.165-172
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    • 2004
  • This study was carried out to examine the effects of maturation media on in vitro maturation (IVM) of porcine immature oocytes, and on subsequent in vitro fertilization (IVF) and development (IVD). Cumulus-oocyte complexes (COCs) were collected from antral follicles of porcine ovaries collected from abattoir, and were matured in vitro in modified NCSU-37 (mNCSU-37), modified NCSU-23 (mNCSU-23), or TCM-199 supplemented with 10% porcine follicular fluid (pFF). Oocytes matured in vitro, were fertilized in vitro in modified Tris-buffered medium(mTBM) with the final motile sperm concentration of 1${\times}$105 sperm/mL, and subsequently putative embryos were developed in vitro in NCSU-23. The results are as follows. 1. In the result of IVM, the rate of germinal vesicle breakdown (GVBD) and of nuclear maturation were not significantly different among the media, though numeric value of them were slightly lower in TCM-199 than in mNCSU-37 or in mNCSU-23. 2. In the result of IVF, though the rate of sperm penetration was not significantly different among the maturation media, the percentage of oocytes with male pronucleus (MPN) of ones matured in mNCSU-37 (88.0%) was significantly higher than in TCM-199 (71.1%) (p<0.05). 3. In the result of IVD, the percentage of cleaved oocytes of ones matured in mNCSU-37 (52.3%) or in mNCSU-23 (53.7%) was significantly higher than in TCM-199 (43.1%) (p<0.05), but the rate of blastocysts at day 6 was not significantly different among the maturation media, though putative embryos from oocytes matured in mNCSU-37 or in mNCSU-23 were developed more than in TCM-199. These results suggested that mNCSU-37 or mNCSU-23 was more appropriate than TCM-199 as IVM medium for porcine immature oocytes.

Sex Determination of Porcine IVF Embryos by Polymerase Chain Reaction (PCR) (중합효소연쇄반응을 이용한 돼지 체외수정란의 성감별)

  • 강미선;김용준;이해이;유일정
    • Journal of Embryo Transfer
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    • v.16 no.3
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    • pp.203-211
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    • 2001
  • This study was carried out to determine sex of porcine embryos produced by in vitro fertilization. Porcine oocyte-cumulus complexes were cultured in BSA-free North Carolina State University (NCSU) 23 medium containing porcine follicular fluid (10%), cystein (0.1 mg/ml) and hormonal supplement (10 IU eCG and 10 IU hCG per ml) for 20~22 hrs. They were then cultured in the same medium but without hormonal supplement for additional 20~22 hrs. After culture, cumulus cells were removed and oocytes were co-incubated for 6 hrs with four different concentrations (5$\times$10$^4$, 2.5$\times$ 10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) of porcine sperm. After fertilization, oocytes were transferred into NCSU 23 with 0.4% BSA medium. The cleavage and blastocyst formation rates were evaluated at 48 and 144 hrs, respectively. In this study, the polymerase chain reaction (PCR) was used to determine the sex of porcine embryos in the stage of blastocyst. The PCR was performed using a set of oligonucleotide primers (5‘-TCATGGACCAGGTAGGGAAT-3', 5’-GAAAGACACGTCCTTGGA GA-3') for 491 bp fragment of porcine male-specific DNA sequence. In the flour different sperm concentration (5$\times$10$^4$, 2.5$\times$10$^{5}$ , 5.0$\times$10$^{5}$ and l0$\times$10$^{5}$ ) for fertilization condition, the cleavage rate was 55.95, 67.88, 60.18 and 47.60%, respectivety, and the development rate of blastocysts was 16.03, 20.40, 21.41 and 12.37%, respectively. At 5.0$\times$10$^4$and 2.5$\times$10$^{5}$ of sperm concentrations per ml cleavage rate and development rate of blastocyst were higher than those of 5.0$\times$10$^4$and l0$\times$10$^{5}$ of sperm concentration (P<0.01). The male of porcine embryos was detected at 491 bp by PCR, and 18 of the 31 porcine blastocysts were the male (58.1%) and the rest 13 were the female(41.9%).

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Height Datum Transformation using Precise Geoid and Tidal Model in the area of Anmyeon Island (정밀 지오이드 및 조석모델을 활용한 안면도 지역의 높이기준면 변환 연구)

  • Roh, Jae Young;Lee, Dong Ha;Suh, Yong Cheol
    • Journal of Korean Society for Geospatial Information Science
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    • v.24 no.1
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    • pp.109-119
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    • 2016
  • The height datum of Korea is currently separated into land and sea, which makes it difficult to acquire homogeneous and accurate height information throughout the whole nation. In this study, we therefore tried to suggest the more effective way to transform the height information were constructed separately according to each height datum on land and sea to those on the unique height datum using precise geoid models and tidal observations in Korea. For this, Anmyeon island was selected as a study area to develop the precise geoid models based on the height datums land (IMSL) and sea (LMSL), respectively. In order to develop two hybrid geoid models based on each height datum of land an sea, we firstly develop a precise gravimetric geoid model using the remove and restore (R-R) technique with all available gravity observations. The gravimetric geoid model were then fitted to the geometric geoidal heights, each of which is represented as height datum of land or sea respectively, obtained from GPS/Leveling results on 15 TBMs in the study area. Finally, we determined the differences between the two hybrid geoid models to apply the height transformation between IMSL and LMSL. The co-tidal chart model of TideBed system developed by Korea Hydrographic and Oceanographic Agency (KHOA) which was re-gridded to have the same grid size and coverage as the geoid model, in order that this can be used for the height datum transformation from LMSL to local AHHW and/or from LMSL to local DL. The accuracy of height datum transformation based on the strategy suggested in this study was approximately ${\pm}3cm$. It is expected that the results of this study can help minimize not only the confusions on the use of geo-spatial information due to the disagreement caused by different height datum, land and sea, in Korea, but also the economic and time losses in the execution of coastal development and disaster prevention projects in the future.