• Title/Summary/Keyword: Two-cell

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Production of cloned Mice by Nuclear Transplantation and Electrofusion Using 2- or 8-Cell Stage Mouse Embryo as Nuclear Donor (2- 및 8- 세포기 생쥐 수정란의 핵이식 및 전기융합법에 의한 복제산자의 생산)

  • 박준규;조성근;박희성;박충생
    • Journal of Embryo Transfer
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    • v.10 no.3
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    • pp.209-217
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    • 1995
  • The present study was carried out to develop a cloning technology of mouse embryos by nuclear transplantation with electrofusion and to produce cloned offsprings by transfer of reconstituted embryos. A single nucleus from two- and eight-cell embryos was transplanted into the enucleated two-cell embryos by rnicromanipulation. The fusion of nucleus with recipient cytoplasm and the subsequent development of reconstituted embryos in vitro as well as in vivo to term were examined to determine the optimal electrofusion parameters for nuclear transplantation in mouse embryos. The successful enucleation of donor embryos was 84.9 and 83.3% in two- and eight-cell stage, respectively, and the successful injection of nucleus from two- and eight-cell donor embryos into the perivitelline space of enucleated two-cell embryos were 85.1 and 84.7%, respectively. No significant differences were found in enucleation or injection rate between the cell stages of donor embryos. When the blastomeres of intact two-cell mouse embryos were electrofused in 0.3 M mannitol medium(100 $\mu$sec., 3 pulses), the fusion rate was similarly 93.2, 92.2 and 92.0% in 1.0, 1.5 and 2.0 kV /crn, respectively, but in vitro development to blastocyst of the fused two-cell embryos was significantly(P<0.05) lower in 2.0 kV/cm (63.4%) than in 1.0 kV/cm (91.7%) or 1.5 kV/cm (82.4%). The development in vitro to eight-cell stage of the reconstituted embryos with nucleus from two-cell stage(45.5%) was significantly(P<0.05) higher than that from eight-cell stage blastomeres (16.7%). The number of blastomeres of the intact embryos at blastocyst stage was 50i0.6 and 55$\pm$2.4 in in vitro and in vivo cultured mouse embryos, respectively, but significantly(P<0.05) decreased to 35$\pm$0.7 in nuclear transplanted blastocyst embryos. The conception rate of mice following embryo transfer was 32.1% in the reconstituted two-cell embryos using two-cell donor nuclei, which was comparable to the fresh two-cell embryos(40.6%). However, the rate of development in vivo to term following embryo transfer of the reconstituted two-cell embryos using two-cell donor nuclei (23.5%) was significantly(P<0.05) lower compared with the percentage of two-cell fresh embryos(31.5%).

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Fabrication and Characteristics of Supported Type Planar Solid Oxide Fuel Cell By Co-firing Process (공소결법에 의해 제조된 지지체식 평판형 고체산화물 연료전지 성능 특성)

  • Song, Rak-Hyun
    • Korean Journal of Materials Research
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    • v.13 no.3
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    • pp.160-168
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    • 2003
  • The co-firing processes for the supported type planar solid oxide fuel cell were investigated. A flat cell of $7.7${\times}$10.8\textrm{cm}^2$ was fabricated successfully by the co-firing process, in which green films were co-sintered in the forms of two layers of anode/electrolyte or of three layers of anode/electrolyte/cathode with gas distributor. A co-fired cell of two layers yielded a power of 200 ㎽/$\textrm{cm}^2$ at 608 ㎷. Its performance loss was mainly due to iR drop in the anodic gas distributor, which was attributed to poor contact between anodic gas distributor and current collector. The performance in the co-fired cell of three layers was much lower than that of two layers, which resulted from the large iR drop and activation overvoltage at the cathodic side. In the co-fired cell of two layers, the impedance analysis indicated that the performance decay during cell operation is due to both anode overvoltage and iR drop at anode side. Also the electrode reaction of the co-fired two layers' cell is considered to be controlled by activation overvoltage within the low current of 50 ㎃.

Improvement of Hydrocarbon Recovery by Two-Stage Cell-Recycle Extraction in the Cultivation of Botryococcus braunii

  • An, Jin-Young;Sim, Sang-Jun;Kim, Byung-Woo;Lee, Jin-Suk
    • Journal of Microbiology and Biotechnology
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    • v.14 no.5
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    • pp.932-937
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    • 2004
  • In situ extraction by organic solvent was studied in order to improve the recovery yield of hydrocarbon from the culture of Botryococcus braunii, a green colonial microalga. When the solvent mixture of octanol as an extractive solvent and n-octane as a biocompatible solvent was added to a two-phase column, the algal growth was seriously inhibited, even at a low concentration of polar octanol. Therefore, a two-stage cell-recycle extraction process was proposed to improve the contact area between the organic phase and the aqueous phase. The hydrocarbon recovery with in situ cell-recycle extraction showed a three-fold increase (57% of cell) in yield over that with two-phase extraction. In addition, over 60% of the hydrocarbon could be recovered without serious cell damage by downstream separation when this process was applied to the culture broth after batch fermentation.

Establishment and Characterization of Permanent Cell Lines from Oryzias dancena Embryos

  • Lee, Dongwook;Kim, Min Sung;Nam, Yoon Kwon;Kim, Dong Soo;Gong, Seung Pyo
    • Fisheries and Aquatic Sciences
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    • v.16 no.3
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    • pp.177-185
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    • 2013
  • The development of species-specific fish cell lines has become a valuable tool for biological research. In recent years, marine medaka Oryzias dancena has been recognized as a good experimental model fish but there are no reports of establishment of cell lines from this fish. In this study, two cell lines from O. dancena blastula embryos were established from 41 total trials (4.9%). The two cell lines displayed typical in vitro morphology and have been cultured for >121 passages, which corresponds to 293 days. The doubling times of the cell lines were 29.84 and 28.59 h, respectively, and both possessed the potential to expand in a clonal manner, albeit with significant differences between the two cell lines. The absence of any of the four main medium supplements; i.e., fish serum, fetal bovine serum, basic fibroblast growth factor, and medaka embryo extract, significantly inhibited growth. The proportion of cells possessing normal chromosome number was 45% and 46.7% of the cell lines, respectively. Taken together, two cell lines that proliferate continuously were established from marine medaka and these cell lines may provide a basic tool for characterizing the unique features of this fish species.

Modeling of two-cell thin-walled beams using variational asymptotic methods (변분적 점근법을 사용한 이중 세포를 갖는 박벽보의 모델링)

  • Park, Jae-Sang;Kim, Ji-Hwan
    • Proceedings of the Korean Society For Composite Materials Conference
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    • 2005.11a
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    • pp.198-201
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    • 2005
  • This study investigates the difference between single-cell and multi-cell cross-sections of thin-walled beams. The variationally and asymptotically consistent theory is used in order to model the two-cell thin- walled beam. The theory is based on an asymptotical analysis of two-dimensional shell energy. In addition, the method allows for the development of closed-form expressions for the displacement, stress field and beam stiffness coefficients. The numerical results show the difference between the cross-sectional stiffness of single-cell and that of multi-cell.

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Study on In Vitro Development of Mouse Embryos (생쥐 수정란의 핵이식후 체외발달에 관한 연구)

  • 박희성;이효종;최상용;박충생
    • Korean Journal of Animal Reproduction
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    • v.14 no.3
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    • pp.205-211
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    • 1990
  • Single nuclei from two-, four- and eight-cell mouse embryos were transplanted into enucleated two-cell mouse embryos by micromanipulation and sendai virum mediated fusion. no significant difference in successful injectin rate and fusion rate was found between the cell stages of nuclear donor embryos. There nuclear transplant embryos receiving different cell stage nuclei were cultured in vitro for 96 hours. 75.3% of 255 embryos receiving 2-cell nuclei, 68.2% of 236 embryos reciving 4-cell nuceli and 46.9% of 228 embryos receiving 8-cell nuclei were developed to blastocyst, respectively. The number of blastomeres was significantly(P<0.05) reduced in the embryos receiving 8-cell nuclei, compared with the embryos receiving 2-cell, 4-cell nuclei or the intact embryos. Also the size of blastocysts was significantly(P<0.05) smaller in the embryos receiving 8-cell nuclei, compared with the intact or other nuclear transplant embryos. These results suggest that single nuclei introduced into the enucleated two-cell embryos are able to support the in vitro development of the reconstituted embryos to blastocysts. The prominant retardation of blastocoele formation and cell division was shown in nuclear transplant embryos receiving eight-cell nuclei when they were cultured in vitro.

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Cell Wall Localization of Two DUF642 Proteins, BIIDXI and TEEBE, during Meloidogyne incognita Early Inoculation

  • Salazar-Iribe, Alexis;Zuniga-Sanchez, Esther;Mejia, Emma Zavaleta;Gamboa-deBuen, Alicia
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.614-618
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    • 2017
  • The root-knot nematode Meloidogyne incognita infects a variety of plants, including Arabidopsis thaliana. During migration, root-knot nematodes secrete different proteins to modify cell walls, which include pectolytic enzymes. However, the contribution of host cell wall proteins has not been described during this process. The function of two DUF642 cell wall proteins, BIIDXI (BDX, At4g32460) and TEEBE (TEB, At2g41800), in plant development could be related to the regulation of pectin methyl esterification status in the cell walls of different tissues. Accordingly, the expression of these two genes is up-regulated by auxin. BDX and TEB were highly induced during early M. incognita inoculation. Moreover, cell wall localization of the proteins was also induced. The cell wall localization of BDX and TEB DUF642 proteins during M. incognita early inoculation suggested that these two proteins could be involved in the regulation of the degree of pectin methylation during cell separation.

Experimental Analyses of Cell Voltages for a Two-cell PEM Stack Under Various Operating Conditions

  • Park, Sang-Kyun;Choi, Jae-Hyuk
    • Journal of Advanced Marine Engineering and Technology
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    • v.35 no.7
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    • pp.881-890
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    • 2011
  • Analyses of performance and behavior of the individual PEM fuel cells (PEMFC) under different operating conditions are of importance optimally to design and efficiently to operate the stack. The paper focuses on experimental analyses of a two-cell stack under different operating conditions, which performance and behavior are measured by the voltage of a cell as well as the stack. Experimental parameters include stoichiometric ratio, temperature of the air supplied under different working stack temperatures and loads. Results showed that the cell voltages are dominantly influenced by the temperature of the air supplied among others. In addition, an inherent difference between the first and the second cell voltage exists because of the tolerances of the cell components and the resulting different over-potentials at different equilibrium states. Furthermore, it is shown that the proton conductivity in the membranes conditioned by the humidity in the cathode channel highly affects the voltage differences of the two cells.

Yeast two-hybrid assay with fluorescence reporter (형광 리포터를 활용한 효모 단백질 잡종 기법 개발)

  • Park, Seong Kyun;Seo, Su Ryeon;Hwang, Byung Joon
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.199-205
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    • 2019
  • Yeast two-hybrid (Y2H) technique has been used to study protein-protein interactions, but its application particularly to a large-scale analysis of protein interaction networks, is limited by the fact that the technique is labor-intensive, based on scoring colonies on plate. Here, we develop a new reporter for the measurement of the protein-protein interactions by flow cytometry. The yeast harboring interacting proteins can also be enriched by fluorescence-activated cell sorting (FACS) or magnetic-activated cell sorting (MACS). When two interacting proteins are present in the same yeast cell, a reporter protein containing 10 tandem repeats of c-myc epitope becomes localized on the surface of the cell wall, without affecting cell growth. We successful measured the surface display of c-myc epitope upon interacting p53 with SV40 T antigen by flow cytometry. Thus, the newly developed Y2H assay based on the display of c-myc repeat on yeast cell wall could be used to the simultaneous analysis of multiple protein-protein interactions without laborious counting colonies on plate.

Two Dimensional Numerical Model for Thermal Management of Proton Exchange Membrane Fuel Cell with Large Active Area (대면적 셀 고분자 막전해질 연료전지의 열관리를 위한 2 차원 수치 해석 모델)

  • Yu, Sang-Seok;Lee, Young-Duk;Ahn, Kook-Young
    • Transactions of the Korean Society of Mechanical Engineers B
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    • v.32 no.5
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    • pp.359-366
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    • 2008
  • A two-dimensional thermal model of proton exchange membrane fuel cell with large active area is developed to investigate the performance of fuel cell with large active area over various thermal management conditions. The core sub-models of the two-dimensional thermal model are one-dimensional agglomerate structure electrochemical reaction model, one-dimensional water transport model, and a two-dimensional heat transfer model. Prior to carrying out the simulation, this study is contributed to set up the operating temperature of the fuel cell with large active area which is a maximum temperature inside the fuel cell considering durability of membrane electrolyte. The simulation results show that the operating temperature of the fuel cell and temperature distribution inside the fuel cell can affect significantly the total net power at extreme conditions. Results also show that the parasitic losses of balance of plant component should be precisely controlled to produce the maximum system power with minimum parasitic loss of thermal management system.