• Title/Summary/Keyword: Nuclear Fusion

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In Vitro Development of Nuclear Transplantation Bovine Embryos Using In Vitro Fertilized Embryos of Korean Native Heifers (한우 체외수정란을 이용한 핵 이식배의 체외발달에 관한 연구)

  • 박충생;공일근;노규진;이효종;최상용
    • Korean Journal of Animal Reproduction
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    • v.18 no.2
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    • pp.113-119
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    • 1994
  • To improve nuclear transplantation(NT) efficiency and to produce a large scale genetically identical cloned calves, examined the in vitro development capacity after co-culture of bovine oviductal epithelial cells (BOEC) and granulosa cells in TCM-199 supplemented with 10% fetal calf serum (FCS) with early bovine embryos derived from in vitro matured fertilized(IVM-IVF) oocyte. In addition, the age dependence of IVM oocyte on electro-stimulation and the effective electric voltage on in ivtro development of bovine NT embryos were examined. The results obtained were summerized as follows; 1. The cleavage rates of IVM-IVF bovine embryos in co-culture with bovine oviductal epithelial cells and granulosa cells were not significantly different(P<0.05), but the developmental rate into morula and blastocyst stage were different showing 38.3 and 20.2%, respectively. 2. The activation (82.5%) and development in vitro(8.6%) into later embryo stages of the aging oocytes of 32 hours post-maturation (hpm) were significantly higher than those of 24 hpm at direct current (DC) voltage of 1.5kV/cm, 60$\mu$sec pulse duration and 1 pulse time. 3. The fusion rates of NT eggs of 32 hpm following to different DC voltages from range 0.75 to 1.5kV/cm were not differ, but the developmental rate into morula and blastocyst stages at DC voltages of 0.75 and 1.0kV/cm were higher(11.4 and 12.6%, respectively) than those of 1.5kV/cm(0%). From these results, it can be suggested the optimal culture system for in vitro culture of IVM-IVF bovine embryos is a co-culture system with BOEC in TCM-199 supplemented 10% FCS. The effective time and the DC voltage for activation, electrofusion and in vitro development of NT embryos derived from IVM-IVF bovine embryo are 32hpm and 0.75~1.0kV/cm. But to improve NT efficiency, the advanced research (cell cycle synchronization, micromanipulation, culture system, etc.) is needed.

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Porcine somatic cell nuclear transfer using telomerase reverse transcriptase-transfected mesenchymal stem cells reduces apoptosis induced by replicative senescence

  • Jeon, Ryounghoon;Rho, Gyu-Jin
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.3
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    • pp.215-222
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    • 2020
  • Mesenchymal stem cells (MSCs) have been widely used as donor cells for somatic cell nuclear transfer (SCNT) to increase the efficiency of embryo cloning. Since replicative senescence reduces the efficiency of embryo cloning in MSCs during in vitro expansion, transfection of telomerase reverse transcriptase (TERT) into MSCs has been used to suppress the replicative senescence. Here, TERT-transfected MSCs in comparison with early passage MSCs (eMSCs) and sham-transfected MSCs (sMSCs) were used to evaluate the effects of embryo cloning with SCNT in a porcine model. Cloned embryos from tMSC, eMSC, and sMSC groups were indistinguishable in their fusion rate, cleavage rate, total cell number, and gene expression levels of OCT4, SOX2 and NANOG during the blastocyst stage. The blastocyst formation rates of tMSC and sMSC groups were comparable but significantly lower than that of the eMSC group (p < 0.05). In contrast, tMSC and eMSC groups demonstrated significantly reduced apoptotic incidence (p < 0.05), and decreased BAX but increased BCL2 expression in the blastocyst stage compared to the sMSC group (p < 0.05). Therefore, MSCs transfected with telomerase reverse transcriptase do not affect the overall development of the cloned embryos in porcine SCNT, but enables to maintain embryo quality, similar to apoptotic events in SCNT embryos typically achieved by an early passage MSC. This finding offers a bioengineering strategy in improving the porcine cloned embryo quality.

Developmental Ability of Bovine Embryos Nuclear Transferred with Frozen-thawed or Cooled Donor Cells

  • Hong, S.B.;Uhm, S.J.;Lee, H.Y.;Park, C.Y.;Gupta, M.K.;Chung, B.H.;Chung, K.S.;Lee, H.T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.9
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    • pp.1242-1248
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    • 2005
  • This study was designed to investigate the in vitro developmental ability and apoptosis of bovine embryos nucleartransferred (NT) with frozen-thawed or cooled donor cells. Cultured adult bovine ear cells were used as donor cells after sub-culturing to confluence (CC), cooling to 4$^{\circ}C$ for 48 h, or freezing-thawing (FT). Apoptotic cells in blastocysts were evaluated for apoptosis by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) method. Fusion, cleavage and blastocyst rates were 69.0 (167/242), 68.8 (115/167), and 29.9 (50/167) with CC cells, 70.4 (88/125), 69.3 (61/88), and 29.6 (26/88) with cooled cells and 66.1 (117/177), 70.1 (82/117), and 13.7 (16/117) with FT cells, respectively. Blastocyst rates of NT embryos derived from FT cells were significantly lower than those from CC or cooled cells (p<0.05). In addition, NT blastocysts produced by using FT cells showed significantly higher apoptosis rates (6.4${\pm}$4.0%) than those produced by CC (2.8${\pm}$1.7%) or cooled (2.3${\pm}$1.3%) cells. However, cooling of donor cells had no significant adverse effect on blastocyst rate as well as apoptosis rate. Therefore, our results suggest that cooled cells may be used as an alternative to freshly cultured confluent culture cells, as donor cells, for the production of Somatic nuclear cloned cattle.

The Effects of Donor Cell Type and Culture Medium on in vitro Development of Domestic Cat Embryos Reconstructed by Nuclear Transplantation

  • Fahrudin, Mokhamad;Otoi, Takeshige;Karja, Ni Wayan Kurniani;Murakami, Masako;Suzuki, Tatsuyuki
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.8
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    • pp.1057-1061
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    • 2001
  • In this study we explored the possibility of performing nuclear transfer in the domestic cat and assessed the ability of different culture media to support in vitro development of reconstructed cat embryos. Donor somatic cells were derived from cultured cumulus cells or explants of oviduct tissue, and recipient cytoplasts from in vitro matured oocytes. A higher percentage of cleavage (84.6% and 86.5%) and development to the morula stage (35.9% and 44.2%) was found when reconstructed embryos receiving cumulus or oviduct cells were cultured in MK1 medium, compared with those cultured in CR1aa (58.7% and 72.5%, 13.8% and 13.6%, respectively). There was no significant difference between MK1 and CR1aa media with respect to the proportion developing to the blastocyst stage (15.4% and 17.3% vs 6.8% and 8.6%, respectively, p>0.05). There was no significant effect (p>0.05) of donor cell type (cumulus and oviduct cells) on the rates of fusion (65.0% and 52.5%), cleavage (84.6% and 86.5%), development to the morula (35.9% and 44.2%), and blastocyst (15.4% and 17.3%) stages when reconstructed embryos were cultured in MK1 medium. Similar results were found for the reconstructed embryos cultured in CR1aa medium. These results show that culture medium has a significant impact on the early development of reconstructed cat embryos, whereas donor cell type does not have a significant effect.

Correlation of Oct4 and FGF4 Gene Expression on Peri-implantation Bovine Embryos Reconstructed with Somatic Cell

  • K. S. Chung;Yoon, B. S;S. J. Song;Park, Y. J.;S. B. Hong;Lee, H. T.
    • Korean Journal of Animal Reproduction
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    • v.26 no.4
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    • pp.329-338
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    • 2002
  • This study was carried out to investigate the developmental rates of embryo reconstructed with different cell type and to estimate correlation of transcriptional level of octamer-binding transcription factor 4 (Oct4) and fibroblast growth factor 4 (FCF4) gene on peri-implantation stage embryos. Donor cells were transferred into perivitelline space of enucleated oocytes. The karyoplast-cytoplast couplets were accom- plished by cell to cell fusion and activated with ionomycin and 6-dimethylaminopurine. Reconstructed embryos were co-cultured with bovine oviduct epithelial cells in CR 1 aa medium. There is no difference in blastocyst formation rate following nuclear transfer UT) with fetal fibroblast cell (16/50; 32.0%), cumulus cell (16/49; 32.6%) and ear cell (17/52; 32.6%). The expression level of Oct4 and FCF4 in peri-implantation bovine embryo derived from in vitro fertilization (IVF) and NT were determined by reverse-transcription polymerase chain reaction (RT-PCR) technique. In peri-implantation of IVF result in a transient increased of FCF4 paralleled by an increased expression of Oct4. However, Oct4 gene was highly expressed in hatching blastocysts derived from NT compared to IVF. Also, FGF4 expression level in hatching blastocysts and outgrowth stage derived from NT was lower than that of IVF. In conclusion, it is suggested that the different transcription patterns observed in nuclear transfer embryos may lead to a lower rate of embryo development, implantation and pregnancy.

Whether Pinhole Scan or Single Photon Emission Computed Tomography (SPECT) in the Diagnosis of Bone and Joint Diseases (골격계진단에 있어서 핀홀스캔의 우월성)

  • Bahk, Yong-Whee
    • The Korean Journal of Nuclear Medicine
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    • v.30 no.1
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    • pp.1-14
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    • 1996
  • Since the publication of the first bone scintiscans in 1962 three decades have elapsed. The bone scan has made great strides during this period, becoming one of the most commonly used nuclear imaging tests. In spite of the progress, however, the specificity of bone scan has remained relatively low. As the result it is a common practice to seek additional information from radiograph, CT scan and MR image, which is euphemistically termed as "image fusion or co-location." The basic reason is the inapplicability of the classical piecemeal analysis to interpreting planar and SPECT bone scans. Such analysis has its base on the observation of elemental features of morphology, which include the size, shape, contour, location, topography and internal architecture. The physiochemical profile may well also be included. Understandably, however, the miniatured images of the planar bone scan cannot provide these features in acceptable detail and the same holds true even with SPECT Images which are but sliced views of the reconstructed planar scans. Fortunately pinhole scanning has the capacity to portray both the morphological and chemical profiles of bone and joint diseases in greater detail through true magnification. The magnitude of pinhole scan resolution is practically comparable to that of radiography as far as gross anatomy is concerned. Thus, we feel strongly that pinhole scanning is a potential breakthrough of the long-lamented low specificity of bone scan. This presentation will discuss the fun-damentals, advantages and disadvantages and the most recent advances of pinhole scanning. It high-lights the actual clinical applications of pinhole scanning in relation to the diagnosis of infective and inflammatory diseases of bone and joint.

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Dynamic characteristics of single door electrical cabinet under rocking: Source reconciliation of experimental and numerical findings

  • Jeon, Bub-Gyu;Son, Ho-Young;Eem, Seung-Hyun;Choi, In-Kil;Ju, Bu-Seog
    • Nuclear Engineering and Technology
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    • v.53 no.7
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    • pp.2387-2395
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    • 2021
  • Seismic qualifications of electrical equipment, such as cabinet systems, have been emerging as the key area of nuclear power plants in Korea since the 2016 Gyeongju earthquake, including the high-frequency domain. In addition, electrical equipment was sensitive to the high-frequency ground motions during the past earthquake. Therefore, this paper presents the rocking behavior of the electrical cabinet system subjected to Reg. 1.60 and UHS. The high fidelity finite element (FE) model of the cabinet related to the shaking table test data was developed. In particular, the first two global modes of the cabinet from the experimental test were 16 Hz and 24 Hz, respectively. In addition, 30.05 Hz and 37.5 Hz were determined to be the first two local modes in the cabinet. The high fidelity FE model of the cabinet using the ABAQUS platform was extremely reconciled with shaking table tests. As a result, the dynamic properties of the cabinet were sensitive to electrical instruments, such as relays and switchboards, during the shaking table test. In addition, the amplification with respect to the vibration transfer function of the cabinet was observed on the third floor in the cabinet due to localized impact corresponding to the rocking phenomenon of the cabinet under Reg.1.60 and UHS. Overall, the rocking of the cabinet system can be caused by the low-frequency oscillations and higher peak horizontal acceleration.

Silencing of NbNAP1 Encoding a Plastidic SufB-like Protein Affects Chloroplast Development in Nicotiana benthamiana

  • Ahn, Chang Sook;Lee, Jeong Hee;Pai, Hyun-Sook
    • Molecules and Cells
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    • v.20 no.1
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    • pp.112-118
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    • 2005
  • It was previously shown that AtNAP1 is a plastidic SufB protein involved in Fe-S cluster assembly in Arabidopsis. In this study, we investigated the effects of depleting SufB protein from plant cells using virus-induced gene silencing (VIGS). VIGS of NbNAP1 encoding a Nicotiana benthamiana homolog of AtNAP1 resulted in a leaf yellowing phenotype. NbNAP1 was expressed ubiquitously in plant tissues with the highest level in roots. A GFP fusion protein of the N-terminal region (M1-V103) of NbNAP1 was targeted to chloroplasts. Depletion of NbNAP1 resulted in reduced numbers of chloroplasts of reduced size. Mitochondria also seemed to be affected. Despite the reduced number and size of the chloroplasts in the NbNAP1 VIGS lines, the expression of many nuclear genes encoding chloroplast-targeted proteins and chlorophyll biosynthesis genes remained unchanged.

GENERATION OF MAGNETIC FIELDS IN COSMOLOGICAL SHOCKS

  • MEDVEDEV MIKHAIL V.;SILVA LUIS O.;FIORE MASSIMILIANO;FONSECA RICARDO A.;MORI WARREN B.
    • Journal of The Korean Astronomical Society
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    • v.37 no.5
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    • pp.533-541
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    • 2004
  • The origin of magnetic fields in the universe remains an outstanding problem in cosmology. We propose that these fields are produced by shocks during the large-scale structure formation. We discuss the mechanism of the field generation via the counter-streaming (Weibel) instability. We also show that these Weibel-generated fields are long-lived and weakly coupled to dissipation. Subsequent field amplification by the intra-cluster turbulence may also take place, thus maintaining the magnetic energy density close to equipartition.

The analysis of bending processes in $Nb_{3}$Sn Cable-in-Conduit Conductors ($Nb_{3}$Sn 도관 케이블 도체의 굽힘변형 해석)

  • 남현일;이호진;박재학
    • Proceedings of the Korea Institute of Applied Superconductivity and Cryogenics Conference
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    • 2002.02a
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    • pp.259-262
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    • 2002
  • Cable-in-Conduit Conductors (CICC) have been used for fabrication of nuclear fusion magnets. Bending process is included in fabrication of superconducting magnet such as termination of conductors. Because of plastic deformation by bending process, there can be a large residual stress and change of shape in bent conductors. Void volume fraction in conductors is also changed by bending process. In this study, Commercial code was used to analyze the bending process at various bending curvature radius. The calculated residual stress of conductors bent at less than about 40cm curvature radius exceeded the allowable stress.

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