• Title/Summary/Keyword: Nuclear fusion

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Isolation and Characterization of ACC Synthase Gene Family in Mung Bean (Vigna radiata L.): Differential Expression of the Three ACC Synthase enes in Response to Auxin and Brassinosteroid

  • Sunjoo Joo;Kim, Woo-Taek
    • Journal of Plant Biotechnology
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    • v.2 no.2
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    • pp.61-71
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    • 2000
  • By screening a cDNA library of auxin-treated mung bean (Vigna radiata L.) hypocotyls, we have isolated two full-length cDNA clones, pVR-ACS6 and pVR-ACS7, for 1-aminocyclopropane-1-carboxylate (ACC) synthase, the rate-limiting enzyme in the ethylene biosynthetic pathway. While PVR-ACS6 corresponds to the previously identified PCR fragment pMBA1, pVR-ACS7 is a new cDNA clone. A comparison of deduced amino acid sequences among auxin-induced ACC synthases reveal that these enzymes share a high degree of homology (65-75%) to VR-ACS6 and VR-ACS7 polypeptides, but only about 50% to VR-ACS1 polypeptide. ACS6 and ACS7 are specifically induced by auxin, while ACS1 is induced by cycloheximide, and to lesser extent by excision and auxin treatment. Results from nuclear run-on transcription assay and RNA gel blot studies revealed that all three genes were transcriptionally active displaying unique patterns of induction by IAA and various hormones in etiolated hypocotyls. Particularly, 24-epibrassinolide (BR), an active brassinosteroid, specifically enhanced the expression of VR-ACS7 by distinct temporal induction mechanism compared to that of IAA. In addition, BR synergistically increased the IAA-induced VR-ACS6 and VR-ACS7 transcript levels, while it effectively abolished both the IAA- and kinetin-induced accumulation of VR-ACS1 mRNA. In light-grown plants, VR-ACS1 was induced by IAA in roots, whereas W-ACS6 in epicotyls. IAA- and BR-treatments were not able to increase the VR-ACS7 transcript in the light-grown tissues. These results indicate that the expression of ACC synthase multigene family is regulated by complex hormonal and developmental networks in a gene- and tissue-specific manner in mung bean plants. The VR-ACS7 gene was isolated, and chimeric fusion between the 2.4 kb 5'-upstream region and the $\beta$-glucuronidase (GUS) reporter gene was constructed and introduced into Nicotiana tobacum. Analysis of transgenic tobacco plants revealed the VR-ACS7 promoter-driven GUS activity at a highly localized region of the hypocotyl-root junction of control seedlings, while a marked induction of GUS activity was detected only in the hypocotyl region of the IAA-treated transgenic seedlings where rapid cell elongation occurs. Although there was a modest synergistic effect of BR on the IAA-induced GUS activity, BR alone failed to increase the GUS activity, suggesting that induction of VR-ACS7 occurs via separate signaling pathways in response to IAA and BR.

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Construction of fat1 Gene Expression Vector and Its Catalysis Efficiency in Bovine Fetal Fibroblast Cells

  • Liu, Boyang;Yang, Runjun;Li, Junya;Zhang, Lupei;Liu, Jing;Lu, Chunyan;Lian, Chuanjiang;Li, Zezhong;Zhang, Yong-Hong;Zhang, Liying;Zhao, Zhihui
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.621-628
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    • 2012
  • The FAT-1 protein is an n-3 fatty acid desaturase, which can recognize a range of 18- and 20-carbon n-6 substrates and transform n-6 polyunsaturated fatty acids (PUFAs) into n-3 PUFAs while n-3 PUFAs have beneficial effect on human health. Fat1 gene is the coding sequence from Caenorhabditis elegans which might play an important role on lipometabolism. To reveal the function of fat1 gene in bovine fetal fibroblast cells and gain the best cell nuclear donor for transgenic bovines, the codon of fat1 sequence was optimized based on the codon usage frequency preference of bovine muscle protein, and directionally cloned into the eukaryotic expression vector pEF-GFP. After identifying by restrictive enzyme digests with AatII/XbaI and sequencing, the fusion plasmid pEF-GFP-fat1 was identified successfully. The pEF-GFP-fat1 vector was transfected into bovine fetal fibroblast cells mediated by Lipofectamine2000$^{TM}$. The positive bovine fetal fibroblast cells were selected by G418 and detected by RT-PCR. The results showed that a 1,234 bp transcription was amplified by reverse transcription PCR and the positive transgenic fat1 cell line was successfully established. Then the expression level of fat1 gene in positive cells was detected using quantitative PCR, and the catalysis efficiency was detected by gas chromatography. The results demonstrated that the catalysis efficiency of fat1 was significantly high, which can improve the total PUFAs rich in EPA, DHA and DPA. Construction and expression of pEF-GFP-fat1 vector should be helpful for further understanding the mechanism of regulation of fat1 in vitro. It could also be the first step in the production of fat1 transgenic cattle.

Desirable Suggestions for Korean Geo-technology R&D through Analysis of the Global Grand Challenges and Moonshot Projects (글로벌 과학난제 도전연구프로젝트 분석을 통한 우리나라 지질자원기술에의 바람직한 제언)

  • Kim, Seong-Yong;Sung, Changmo
    • Economic and Environmental Geology
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    • v.53 no.1
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    • pp.111-120
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    • 2020
  • Remarkable scientific and technological achievements are mainly shown in the 'super-convergence' or 'convergence of convergence' among cross- disciplinary fields, and advanced countries are promoting the 'high-risk, high-return research' ecosystem. Google LLC is carrying out numerous new challenges in terms of a non-failure perspective. Innovative research by the US Defense Advanced Research Projects Agency (DARPA) has produced such breakthroughs as the Internet, GPS, semiconductors, the computer mouse, autonomous vehicles, and drones. China is pioneering a 'Moon Village' and planning the world's largest nuclear fusion energy and ultra-large particle accelerator project. Japan has also launched 'the moonshot technology development research system' to promote disruptive innovation. In Korea, the government is preparing a new research program to tackle the global scientific challenges. Therefore, it is necessary to determine the reasonable geoscientific challenges to be addressed and to conduct a preliminary study on these topics. For this purpose, it is necessary to conduct long-term creative research projects centered on young researchers, select outstanding principal investigators, extract innovative topics without prior research or reference, simplify research proposal procedures, innovate the selection solely based on key ideas, and evaluate results by collective intelligence in the form of conferences.

Development of the Automated Ultrasonic Testing System for Inspection of the flaw in the Socket Weldment (소켓 용접부 결함 검사용 초음파 자동 검사 장비 개발)

  • Lee, Jeong-Ki;Park, Moon-Ho;Park, Ki-Sung;Lee, Jae-Ho;Lim, Sung-Jin
    • Journal of the Korean Society for Nondestructive Testing
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    • v.24 no.3
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    • pp.275-281
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    • 2004
  • Socket weldment used to change the flow direction of fluid nay have flaws such as lack of fusion and cracks. Liquid penetrant testing or Radiography testing have been applied as NDT methods for flaw detection of the socket weldment. But it is difficult to detect the flaw inside of the socket weldment with these methods. In order to inspect the flaws inside the socket weldment, a ultrasonic testing method is established and a ultrasonic transducer and automated ultrasonic testing system are developed for the inspection. The automated ultrasonic testing system is based on the portable personal computer and operated by the program based Windows 98 or 2000. The system has a pulser/receiver, 100MHz high speed A/D board, and basic functions of ultrasonic flaw detector using the program. For the automated testing, motion controller board of ISA interface type is developed to control the 4-axis scanner and a real time iC-scan image of the automated testing is displayed on the monitor. A flaws with the size of less than 1mm in depth are evaluated smaller than its actual site in the testing, but the flaws larger than 1mm appear larger than its actual size on the contrary. This tendency is shown to be increasing as the flaw size increases. h reliable and objective testing results are obtained with the developed system, so that it is expected that it can contribute to safety management and detection of repair position of pipe lines of nuclear power plants and chemical plants.

Effect of Electric Stimulation on Parthenogenesis of In Vitro Matured Oocytes from Korean Native Cows (한우 체외성숙란의 단위발생에 대한 전기자극의 효과)

  • 노규진;공일근;곽대오;이효정;최상용;박충생
    • Journal of Embryo Transfer
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    • v.9 no.2
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    • pp.145-152
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    • 1994
  • The suitable electric stimulation is essential for activation and fusion of oocytes before or after nuclear transplantation The present study was undertaken to determine the optirnal condition for the parthenogenetic activation of in vitro rnatured(IVM) bovine oocytes by electric stimulation. Different direct current(DC) electric voltage of 1.0, 1.5 and 2.0 kV/cm and pulse duration of 30, 60 and 120 $\mu$sec were applied to the JVM nocytes in 0.3 M mannitol solution containing each 100 $\mu$M CaCl$_2$ and MgCl$_2$. IVM occytes at 24, 28 and 32 hours Post-maturation(hpm) were also electrically stimulated at 1.5 kV /cm, for 60 $\mu$ sec. The stimulated nocytes were then co-cultured in TCM-199 solution containing 10% fetal calf serum with bovine oviductal epithelial cells for 7~9 days in a 5% $CO_2$ incubator at 39$^{\circ}C$ ~ Their activation and in vitro development to morula and blastocyst were assessed under an inverted microscope. The higher activation rates 62.8 and 63.4% and in vitro de- velopment rates to morula and blastocyst 5.1 and 10.9% were shown in the oocytes stimulated at the voltage of 1.0 and 1.5 kV/cm than 2.0 kV/cm, respectively. No signifi- cantly(P<0.05) different activation rate was shown in JVM oocytes stimulated for 30, 60 and 120 $\mu$sec, but developmental rates to morula and blastocyst was significantly(P<0.05) higher in the oocytes stimulated for 30 $\mu$sec(6~3%) and 60 $\mu$sec(10~0%) than 120 $\mu$sec(0~ 0%). The aged oocytes at 28 and 30 hpm showed significantly(P<0.05) higher activation rates(72~7 and 79.7%) than the oocytes at 24 hpm(50~9%)~ Also, their developmental rates to morula and blastocyst were significantly(P<0.05) higher in the nocytes at 28(14.3%) and 32 hpm(15.9%) than 24 hpm(3.6%). From these results, it can be suggested that the optimal electric stimulation for IVM bovine occytes is a DC voltage between 1.0 and 1.5 kV/cm, pulse duration of 30 or 60 $\mu$sec, and the optimal age of IVM oocytes for electric activation is at 32 hpm.

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Sampling-based Approach for Seismic Probabilistic Risk Assessment (지진 확률론적 리스크 평가를 위한 샘플링기반 접근법)

  • Kwag, Shinyoung;Eem, Seunghyun;Park, Junhee;Choi, In-Kil
    • Journal of the Computational Structural Engineering Institute of Korea
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    • v.33 no.2
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    • pp.129-136
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    • 2020
  • In this study, we develop a sampling-based seismic probabilistic risk assessment (SPRA) quantification technique that can accurately consider a partially dependent condition of component seismic fragility information. Specifically, the SPRA quantification method is proposed by combining the advantages of two representative methodologies: EPRI seismic fragility and JAERI seismic fragility input-based quantification. The most important feature of the proposed method is that it performs a SPRA using a sampling technique by transforming the EPRI seismic fragility input into JAERI seismic fragility input. When the proposed sampling-based approach was applied to an example of simple system and to a SPRA problem of a nuclear power plant, it was observed that the proposed method yields approximately similar system seismic fragility and seismic risk results as those of the exact solution. Therefore, it is believed that the approach proposed in this study can be used as a useful tool for accurately assessing seismic risks, considering the partial seismic dependence among the components; the existing SPRA method cannot handle such partial dependencies.

Psoraleae Semen Ethanol Extract Inhibits RANKL-Induced Osteoclast Differentiation and Osteoclast Specific Genes Expression (보골지 추출물이 파골세포 분화 및 골흡수 관련 유전자 발현에 미치는 영향)

  • Ryu, Gwang-hyun;Kim, Eom Ji;Kim, Minsun;Kim, Jae-Hyun;Lee, Yujin;Jin, Dae-hwan;Sohn, Youngjoo;Jung, Hyuk-Sang
    • Korean Journal of Acupuncture
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    • v.38 no.3
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    • pp.140-150
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    • 2021
  • Objectives : The increase of osteoclasts could cause osteoporosis and bone-related diseases. Also, the inhibition of osteoclast differentiation is important in treating bone-related diseases. Traditionally, Psoraleae Semen has been used for geriatric diseases, aging and musculoskeletal diseases. The purpose of this study is to investigate the effect of Psoraleae Semen ethanol extract (PS) on osteoclast differentiation and its function. Methods : To confirm the effect of PS on osteoclastogenesis and bone resorption activity, various levels of concentrations of PS (5, 10, 20 and 40 ㎍/ml) were tested on RAW 264.7 cells cultured with RANKL. We measured tartarate-resistant acid phosphatase (TRAP)-positive cells, TRAP activity, pit formation and F-actin ring formation. The expressions of nuclear factor of activated T-cells (NFATc1) and c-Fos were confirmed through western blot and reverse transcription- polymerase chain reaction (RT-PCR). Also, the expression of bone resorption and fusion-related genes in osteoclast was confirmed by RT-PCR. Results : PS decreased the number of TRAP-positive cells and the TRAP activity. In addition, PS significantly inhibited the formation of pit and F-actin ring. Furthermore, PS decreased the expression of osteoclast related genes. Conclusions : PS inhibits osteoclast differentiation and bone resorption ability through inhibition of the expression of osteoclast-related genes. This indicates that PS may be a potential therapeutic agent to osteoporosis by suppressing osteoclastogenesis.

20(S)-ginsenoside Rh2 induces caspase-dependent promyelocytic leukemia-retinoic acid receptor A degradation in NB4 cells via Akt/Bax/caspase9 and TNF-α/caspase8 signaling cascades

  • Zhu, Sirui;Liu, Xiaoli;Xue, Mei;Li, Yu;Cai, Danhong;Wang, Shijun;Zhang, Liang
    • Journal of Ginseng Research
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    • v.45 no.2
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    • pp.295-304
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    • 2021
  • Background: Acute promyelocytic leukemia (APL) is a hematopoietic malignancy driven by promyelocytic leukemia-retinoic acid receptor A (PML-RARA) fusion gene. The therapeutic drugs currently used to treat APL have adverse effects. 20(S)-ginsenoside Rh2 (GRh2) is an anticancer medicine with high effectiveness and low toxicity. However, the underlying anticancer mechanisms of GRh2-induced PML-RARA degradation and apoptosis in human APL cell line (NB4 cells) remain unclear. Methods: Apoptosis-related indicators and PML-RARA expression were determined to investigate the effect of GRh2 on NB4 cells. Z-VAD-FMK, LY294002, and C 87, as inhibitors of caspase, and the phosphatidylinositol 3-kinase (PI3K) and tumor necrosis factor-α (TNF-α) pathways were used to clarify the relationship between GRh2-induced apoptosis and PML-RARA degradation. Results: GRh2 dose- and time-dependently decreased NB4 cell viability. GRh2-induced apoptosis, cell cycle arrest, and caspase3, caspase8, and caspase9 activation in NB4 cells after a 12-hour treatment. GRh2-induced apoptosis in NB4 cells was accompanied by massive production of reactive oxygen species, mitochondrial damage and upregulated Bax/Bcl-2 expression. GRh2 also induced PML/PML-RARA degradation, PML nuclear bodies formation, and activation of the downstream p53 pathway in NB4 cells. Z-VAD-FMK inhibited caspase activation and significantly reversed GRh2-induced apoptosis and PML-RARA degradation. GRh2 also upregulated TNF-α expression and inhibited Akt phosphorylation. LY294002, an inhibitor of the PI3K pathway, enhanced the antitumor effects of GRh2, and C 87, an inhibitor of the TNF-α pathway, reversed NB4 cell viability, and GRh2-mediated apoptosis in a caspase-8-dependent manner. Conclusion: GRh2 induced caspase-dependent PML-RARA degradation and apoptosis in NB4 cells via the Akt/Bax/caspase9 and TNF-α/caspase8 pathways.

Safety evaluation of atmospheric pressure plasma jets in in vitro and in vivo experiments

  • Lee, Ji-Yoon;Park, Shin-Young;Kim, Kyoung-Hwa;Yoon, Sung-Young;Kim, Gon-Ho;Lee, Yong-Moo;Seol, Yang-Jo
    • Journal of Periodontal and Implant Science
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    • v.51 no.3
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    • pp.213-223
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    • 2021
  • Purpose: The atmospheric pressure plasma jet (APPJ) has been introduced as an effective disinfection method for titanium surfaces due to their massive radical generation at low temperatures. Helium (He) has been widely applied as a discharge gas in APPJ due to its bactericidal effects and was proven to be effective in our previous study. This study aimed to evaluate the safety and effects of He-APPJ application at both the cell and tissue levels. Methods: Cellular-level responses were examined using human gingival fibroblasts and osteoblasts (MC3T3-E1 cells). He-APPJ was administered to the cells in the experimental group, while the control group received only He-gas treatment. Immediate cell responses and recovery after He-APPJ treatment were examined in both cell groups. The effect of He-APPJ on osteogenic differentiation was evaluated via an alkaline phosphatase activity assay. In vivo, He-APPJ treatment was administered to rat calvarial bone and the adjacent periosteum, and samples were harvested for histological examination. Results: He-APPJ treatment for 5 minutes induced irreversible effects in both human gingival fibroblasts and osteoblasts in vitro. Immediate cell detachment of human gingival fibroblasts and osteoblasts was shown regardless of treatment time. However, the detached areas in the groups treated for 1 or 3 minutes were completely repopulated within 7 days. Alkaline phosphatase activity was not influenced by 1 or 3 minutes of plasma treatment, but was significantly lower in the 5 minute-treated group (P=0.002). In vivo, He-APPJ treatment was administered to rat calvaria and periosteum for 1 or 3 minutes. No pathogenic changes occurred at 7 days after He-APPJ treatment in the He-APPJ-treated group compared to the control group (He gas only). Conclusions: Direct He-APPJ treatment for up to 3 minutes showed no harmful effects at either the cell or tissue level.

Combined Inland-River Operation Technique for Reducing Inundation in Urban Area: The Case of Mokgam Drainage Watershed (도시지역의 침수저감을 위한 내외수 연계 운영 기법 개발: 목감천 유역을 중심으로)

  • Kwon, Soon Ho;Jung, Hyun Woo;Hwang, Yoon Kwon;Lee, Eui Hoon;Kim, Joong Hoon
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.22 no.1
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    • pp.257-266
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    • 2021
  • Urban areas can often suffer flood damage because of the more frequent catastrophic rainfall events from climate change. Flood mitigation measures consist of (1) structural and (2) non-structural measures. In this study, the proposed method focused on operating an urban drainage system among non-structural measures. The combined inland-river operation technique estimates the inflow of pump stations based on the water level obtained from a preselected monitoring point, and the pump station expels the stored rainwater to the riverside based on those estimates. In this study, the proposed method was applied to the Mokgam drainage watershed, where catastrophic rainfall events occurred (i.e., 2010- and 2011-years), and severe flood damage was recorded in Seoul. Using the proposed method, the efficiency of flood reduction from the two rainfall events was reduced by 34.9 % and 54.4 %, respectively, compared to the current operation method. Thus, the proposed method can minimize the flood damage in the Mokgam drainage watershed by reserving the additional storage space of a reservoir. In addition, flooding from catastrophic rainfall can be prevented, and citizens' lives and property in urban areas can be protected.