• Title/Summary/Keyword: Reproduction experiments

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Studies on Nuclear Transplantation in Mouse Embryos III. Production of Nuclear Transplanted Mice (생쥐배의 핵치환에 관한 연구 III. 핵치환 생쥐의 생산)

  • 박용석;정형민;박세필;이상진;정병현;정길생
    • Korean Journal of Animal Reproduction
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    • v.13 no.2
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    • pp.93-97
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    • 1989
  • These experiments were carried out to develop the technique of nuclear transplantation necessary for elevating utilization efficiency of high quality embryos and the production of clone animals. Embryos of pronucleus stages were obtained from ICR mice. Removal of pronuclei and their transfer to recipient embryos were carried out by micromanipulation and virus mediation. The results obtained in these experiments were summarized as follows ; 1. Total 337 pairs of pronuclear stage embryos were subjected to nuclear transplantatin and 247 pairs(73.3%) of them were successfully transplanted and the number of fused embryos between transplanted nucleus and cytoplasm was 188 pairs(55.8%). 2. Of the 188 fused embryos cultured in vitro, 174(92.4%), 131(69.7%) and 117(62.2%) embryos were developed to 2-cell, morula and blastocyst stages, respectively. 3. When total 104 nuclear transplanted embryos were transferred to uteri of recipient mice on day 2-3 of pseudopregnancy, 4 of 12 recipient mice were pregnant and the number of embryos developed to young was 28(26.9%).

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Separation of X- and Y-Bearing Spermatozoa III. Separation of bull spermtozoa by Sephadex Gel Filtration (X-정자와 Y-정자의 분이에 관한 연구 III. Sephadex Gel 여과에 의한 우정자의 분이)

  • 이주영;엄기붕;고대환;김종배;정길생
    • Korean Journal of Animal Reproduction
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    • v.12 no.1
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    • pp.24-30
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    • 1988
  • These experiments were carried out to develop new techniques for In Vitro separatin of X-and Y-bearing spermatozoa. The bull semen was applied to the various Gel-Columns filled with swellen Sephadex G-50 Fine and then elutriated wtih Locke solution (elutriation rate; 1ml/3-4min., 1ml/1-2min.). Elutriated solution was fractionated into 1ml by automatic Fraction Collector and spermatozoa included in each fraction were subjected to the estimation of viability and recovery rate, and to B-body test. The results obtained in these experiments were summarized as follows: 1. When the column size and the elutriation rate were adjusted to 15$\times$1.6cm and 1ml/3-4min., respectively, the highest sperm concentration was obtained from the 8th to the 12th fraction. 2. As a trend, the viability of spermatozoa was improved by chromatography, and the degree of improvement ranged 5 to 10 percentage. 3. The average recovery rate of spermatozoa applied to column was 73.2 percentage and ranged 52.6 to 81.3 percentage. 4. The lowest rate of B-body bearing spermatozoa following chromatography was obtained when the column size and the elutriation rate were adjusted to 15$\times$0.8cm and 1ml/1-2min., respectively.

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In Vitro Fertilization and Development of Mouse Eggs (생쥐난자의 시험관내 수정과 발달)

  • 김승재;정길생
    • Korean Journal of Animal Reproduction
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    • v.8 no.2
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    • pp.110-115
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    • 1984
  • These experiments were carried out to obtain the information about the optimal pH osmolality affecting in vitro fertilization of the mouse eggs, to elucidate the 2-cell block to development in vitro and to find out the method of controlling the subsequent embryo development in vitro. pH and osomlality was adjusted by adding NaCl or NaHCO3 to the basic salt solution. In vitro fertilization were carried out by inroducting the cumulus masses to the suspension of epididymal spermatozoa at each pH, osmolality, and 10${\mu}$M-EDTA medium. The results obtained in these experiments were summarized as follows: 1. The fertilization rates in vitro at each medium of 235, 252, 269, 286, 306, 323, 345, 368, 393 mosmol were 15.6, 38.2, 65.7, 75.6, 80.9, 74.3, 58.1, 35.1, 24.3, 11.1%, respectively. 2. The fertilization rates in vitro at each medium of pH 6.1, 6.4, 6.7, 7.0, 7.3, 7.6, 7.9, 8.1 were 11.8, 17.9, 32.4, 61.9, 79.5, 76.7, 53.5, 13.6%, respectively. 3. In case of ICR female x ICR male embryos, the development rate of 2-cell embryos to 4-8 cell embryos was 16.2% at normal medium, but the rate was increased to 49.3% in medium containging 10 ${\mu}$M-DETA; In case of C3H female x ICR male embryos, the development rate was 41.0% at normal medium, but the rate was increased to 71.7% at 10 ${\mu}$M-EDTA-medium.

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Studies on the Characteristics of Anti-Zone Antibody I. Identification of Anti-Zona Antibody by Indirect ELISA and Indirect Immunofluorescence Test (항투명대 항체의 특성에 관한 연구 I. 간접효소면역분석법과 간접면역형광분석법에 의한 항투명대 항체의 확인)

  • 김은영;유영춘;박세필;정형민;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.107-114
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    • 1990
  • These experiments were undertaken as a basic study to develop immunocontraceptive vaccine and to understand the role of zona pellucidae in early fertilization process by identifying the monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae by indirect ELISA and indirect immunofluorescence test. The results obtained in these experiments were summarized as follows : 1. The titer of the antibodies to zona was determined by indirect ELISA using solubilized porcine zona coated plates. Both monoclonal and rabbit polyclonal antibodies showed very high titers ; O.D at 1 : 12,800 dilution of antibodies was still significantly higher than that of non-immunized control serum. Rabbit anti-mouse zona pellucidae sera also reacted with porcine zona pellucidae. 2. By indirect immunofluorescence test strong fluorescences were observed on the egg treated with homologous and heterologous rabbit polyclonal antibodies and FITC lablled 2nd antibodies and found to crossreact strongly with the eggs from the pig and mouse. While weaken fluorescences were observed on the eggs treated with monoclonal antibodies.

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Studies on the Characteristics of Anti-Zona Antibody III. Examination of Biochemical Characteristics of Porcine Zone Pellucidae and Anti-Zona Antibody (항투명대 항체의 특성에 관한 연구 III. 돼지투명대와 항투명대 항체의 생화학적 특성 검토)

  • 김은영;박세필;신경순;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.125-131
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    • 1990
  • These experiments were carried out to investigate of the molecular characteristics of porcine zona pellucidae and to examine the reactivity of anti-zona antibody by SDS-PAGE, Immunoblotting and Immunoprecipitation. The results obtained in these experiments were summarized as follows : 1. The result obtained by SDS-PAGE of porcine zona pellucidae indicated that it composed of several units with molecular weight ranging 55,000-110,000. 2. In order to see the reactivity of antibodies to zona pellucidas, immunoblotting was applied. The results indicated that polyclonal antibodies to porcine and mouse zona reacted with porcine zona. While monoclonal antibody to porcine did not react with the procine zona enough to show a clear band on a gel. 3. Labelling of porcine zonae with 125I was performed using the Iodogen method, the radioactivity and the percent incorporation of 125I into porcine zonae were approximately 26,000 cpm/10${mu}ell$ and 16, respectively. 4. Measurements of radioactivity and O.D value for Immunoprecipitates produced by reaction of 125I-porcine zona with anti-zona antisera were confirmed that existence of reactivity of monoclonal antibody to porcine zona although its reactivity was lower than that of polyclonal antibodies, and reconfirmed that cross-reactivity of polyclonal antibody of mouse zona with porcine zona.

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Studies on the Characteristics of Anti-Zona Antibody II. Effect of Anti-Zona Antibody on Fertilization of Porcine and Mouse Eggs In Vitro (항투명대 항체의 특성에 관한 연구 II. 항투명대 항체가 돼지난자와 생쥐난자의 체외수정에 미치는 영향)

  • 김은영;박세필;정형민;정길생;김종배
    • Korean Journal of Animal Reproduction
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    • v.14 no.2
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    • pp.115-124
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    • 1990
  • These experiments were undertaken as a basic study to develop immunocontraceptive vaccine and to understand the role of zona pellucidae in early fertilization process by investigating the effect of monoclonal and polyclonal antibody to porcine zona pellucidae and polyclonal antibody to mouse zona pellucidae on the fertilization of porcine and mouse eggs in vitro. The results obtained in these experiments were summarized as follows : 1. Treatment of porcine and mouse eggs with undiluted anti-zona serum produced intense precipitation layer on the poricne and mouse zonae, respectively, thus resulting in the total inhibition of sperm adherence on surface of zona. 2. In vitro fertilization of eggs pre-treated with 0.3∼10% of various antibodies was examined, and resulting in that 5 and 10% of rabbit polyclonal antibodies to porcine zona inhibited completely both in vitro fertilization and polyspermy of porcine eggs while monoclonal to porcine zona and rabbit polyclonal antibody to mouse zona did not inhibit in vitro fertilization but monoclonal antibody reduced the rate of polyspermy compared to that of control group. Almost the same results were obtained in the study on the effect of anti-zona serum on in vitro fertilization of mouse eggs.

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In Vitro Aggregation and Culture of Mouse Embryos (생쥐 배의 시험관내 응집과 배양)

  • 이상진;정길생
    • Korean Journal of Animal Reproduction
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    • v.8 no.1
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    • pp.29-35
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    • 1984
  • These experiments were carried out to obtain basic information necessary for in vitro culture of aggregated mouse embryos. Inbred ICR mice were used to obtain embryos. The zona pellucida was removed by placing the embryos in Whittingham's medium containing 0.5% protease for about 5-10minutes at 37$^{\circ}C$. Total 263 pairs of 2-, 4- and 8-cell zona free mouse embryos were subjected to aggregation by physical pressure and cultured in Whittingham's medium under the gas phase of 5% CO2 in air at 37$^{\circ}C$ for 24 to 60 hours. The results obtained in these experiments were summarized as follows: 1. Time needed for fusion of 2-, 4- and 8-cell embryos were 0-3, 0-3 and 0-3 hours, respectively and average time needed for in vitro development of 2-, 4- and 8-cell embryos after aggregation to morula and blastocyst were 42, 30 and 13.5 hours, and 51, 39 and 27 hours, respectively. 2. Of total 263 pairs of naked embryos, 227 were firmly aggregated together and the rats of aggregation in 2-, 4- and 8-cell embryos were 71.8, 88.3 and 97.0%, respectively. 3. The rates of aggregated pairs which obtained from 2-, 4- and 8-cell embryos developed to morula were 96.7, 95.6 and 96.9%, respectively, and embryos developed to blastocysts were 88.5, 89.7 and 90.8%, respectively. 4. Conspicuous differences in size of volume and inner cell masses between single and double blastocysts were observed. Although a single blastocolic cavity was formed in most double blastocysts, several formed two distinct cavities from the very beginning.

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Studies on Bisection of Bovine Embryos and Embryo Tranfer (소 우정란의 Bisection과 이식에 관한 연구)

  • 정병현;지희준;이상진;이동희;정태영;정길생
    • Korean Journal of Animal Reproduction
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    • v.13 no.3
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    • pp.164-170
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    • 1989
  • These experiments were carried out to develop the practical technique for the production of identical twins in cattle. Morula and blastocyst stage embryos collected from superovulated donors were bisected into halves by micromanipulation. The resulting demi-embryos were transferred to the uterine horn ipsilateral to the corpus luteum of synchronous recipients. The viability of demi-embryos after splitting was also evaluated by culturing demi-embryos with and without a zona-pellucida. The results obtained in these experiments were summarized as follows : 1. Of total 132 embryos collected by superovulation from 29 donors, 37 embryos were morular and 30 at blastocyst stages. 2. Total 111 demi-embryos were produced from 67 embryos by bisection and 98% of those were normal in morphology. 3. The viability of the demi-embryos cultured with zona-pellucida ranged from 70 to 76.5% and that of the demi-embryos without from 53.8 to 69.2%. 4. The viability of demi-embryos obtained from morula was 63.6% and that of demi-embryos from blastocyst was 73.3%, respectively. 5. 35 demi-embryos were transferred to 21 recipients, 7 of which were confirmed to be pregnant by rectal palpation at 55∼60 days after embryo transfer. One of them produced a calf and 6 are still on pregnancy.

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A Spatial Audio System Using Multiple Microphones on a Rigid Sphere

  • Lee, Tae-Jin;Jang, Dae-Young;Kang, Kyeong-Ok;Kim, Jin-Woong;Jeong, Dae-Gwon;Hamada, Hareo
    • ETRI Journal
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    • v.27 no.2
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    • pp.153-165
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    • 2005
  • The main purpose of a spatial audio system is to give a listener the same impression as if he/she were present in a recorded environment. A dummy head microphone is generally used for such purposes. Because of its human-like shape, we can obtain good spatial sound images. However, its shape is a restriction on its public use and it is difficult to convert a 2-channel recording into multi-channel signals for an efficient rendering over a multi-speaker arrangement. In order to solve the problems mentioned above, a spatial audio system is proposed that uses multiple microphones on a rigid sphere. The system has five microphones placed on special points of the rigid sphere, and it generates audio signals for headphone, stereo, stereo dipole, 4-channel, and 5-channel reproduction environments. Subjective localization experiments show that front/back confusion, which is a common limitation of spatial audio systems using the dummy head microphone, can be reduced dramatically in 4-channel and 5-channel reproduction environments and can be reduced slightly in a headphone reproduction.

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A DIRECT INVERSE-BASED CROSS-TALK CANCELLATION METHOD FOR STEREO AUDIO SYSTEMS (직접 역필터 설계법을 이용한 스테레오 재생시스템의 Cross-talk 제거)

  • Kim, Sang-Myeong;Dogeun Han;Semyung Wang
    • Proceedings of the Korean Society for Noise and Vibration Engineering Conference
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    • 2002.05a
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    • pp.559-564
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    • 2002
  • Cross-talk cancellation, inverse filter design or deconvolution in a generic term, is a vital process for a virtual sound realization in the stereo sound reproduction system. Most, if not all, of the design algorithms available for the inverse filter are based on a linearized model of the real physical plant. The result of such a plant-based design method, which may be referred to here as the indirect method, is biased due to both modelling and inversion errors. This paper presents a novel direct cross-talk cancellation method that may be free from the inversion error. The direct method can directly models the inverse filter by a suitable rearrangement of the input and output ports of the original plant so that no inversion is required here. Advantages are discussed with various experiments in an anechoic chamber using a PC soundcard. Binaural reproduction tests conducted showed that the conventional indirect method yields about 8 % reproduction performance error on both ear positions, whereas the direct method offers about 3 %.

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