• Journal of Plant Biotechnology
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• 제2권2호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제25권5호
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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.

• 자원환경지질
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• 제53권1호
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• pp.111-120
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• 2020

괄목할만한 과학기술성과는 이종분야간 초융합 또는 융합의 융합에서 주로 나타나고 있으며, 기술경쟁국들은 고위험 고수익 연구생태계를 추진하고 있다. 구글은 실패를 두려워하지 않는 관점에서 수많은 새로운 도전을 수행하고 있고, 미국 고등연구계획국(DARPA)의 혁신적인 연구추진은 지금까지 인터넷, GPS, 반도체, 마우스, 자율주행차, 드론 등과 같은 획기적인 성과물을 만들었다. 중국은 달기지 개척, 세계 최대 규모의 핵융합 에너지와 초대형 입자가속기 프로젝트를 추진 중이며, 일본도 '문샷형 기술개발연구제도'를 착수하여 파괴적 혁신창출을 추진하려 한다. 우리나라도 글로벌 과학난제의 도전과 해결을 위해 신규 연구프로그램을 준비하고 있다. 따라서 지질자원기술 연구자가 중심이 되어 도전하고 해결하며 공감할 수 있는 과학난제의 주제 발굴과 이들 주제에 관한 사전연구 등이 필요하다. 이를 위해서는 신진연구자 중심의 장기 창의연구사업, 우수한 과제책임자 선정, 선행연구나 참고문헌도 없는 획기적 주제 도출, 과제제안절차 간소화, 아이디어 중점 선정시스템 혁신, 컨퍼런스 형태의 집단지성에 의한 결과평가 등이 필요하다고 사료된다.

• 비파괴검사학회지
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• 제24권3호
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• pp.275-281
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• 2004

대구경 배관에서 소구경 배관으로 연결하는 부위 및 배관의 방향 전환을 위해 사용하는 소켓용접부는 용입불량 및 사용시간이 경과되면서 내부로부터 진전될 수 있는 균열 등의 결함을 가질 수 있다. 그러나 지금까지 적용하고 있는 비파괴검사법인 액체침투탐상과 방사선투과검사로는 내부에 존재하는 균열성 결함의 검출이 어렵다. 본 연구에서는 소켓용접부 내부의 균열성 결함 검출을 위한 초음파 검사 기법을 확립하였고, 검사를 수행할 수 있는 초음파 탐촉자를 설계 제작하였으며, 자동으로 검사할 수 있는 검사 장비와 제어용 운영 프로그램을 개발하였다. 개발된 장비는 컴퓨터를 기반으로 하고 있으며, pulser/receiver를 내장하고 100 MHz 고속 A/D board를 사용하여 초음파 탐상기 역할을 프로그램으로 구현하였으며, ISA interface type으로 4축 제어용 motion controller를 개발하여 자동 검사를 수행하는 scanner를 제어하도록 하였으며, 검사 결과는 소켓용접부 단면을 실시간 영상으로 나타내도록 되어 있다. 인공결함 시험편의 결함을 평가한 결과 결함의 깊이가 1mm인 결함의 평가 길이는 실제 크기보다 작게 나타내고 있으며, 결함의 깊이가 증가할수록 결함의 길이가 더 크게 평가되었다. 본 연구로 개발한 장비는 원자력 발전소나 화학플랜트에 많이 널려 있는 소켓용접부 내의 용입불량, 피로 균열 등을 검출하여 객관적인 검사 결과를 제시할 수 있으므로 설비 안전 관리 및 보수 부위 결정에 기여할 수 있을 것으로 기대된다.

• 한국수정란이식학회지
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• 제9권2호
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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.

• 한국전산구조공학회논문집
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• 제33권2호
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• pp.129-136
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• 2020

본 연구에서 기기 지진취약도 정보의 부분 종속 관계를 정확하게 고려할 수 있는 샘플링 기반 지진 확률론적 리스크 평가 정량화 기법을 개발하였다. 구체적으로 두 가지 대표적 방법론인 EPRI 지진취약도 입력기반 정량화 방법과 JAERI 지진취약도 입력기반 정량화 기법의 장점을 결합하여 리스크 정량화 방법를 제안하였다. 본 연구에서 제안하는 방법의 가장 중요한 특징은 EPRI 지진취약도 입력을 JAERI 지진취약도 입력 공간으로 치환하여 샘플링 방법으로 SPRA 수행하는 것에 있다. 제안된 샘플링기반 접근법을 간단한 예제부터 실제 원전의 지진 확률론적 리스크 평가 문제에 적용한 결과, 본 연구에서 제안하는 방법이 정해에 가까운 시스템 지진취약도 및 지진리스크 값을 산출함을 확인할 수 있었다. 그러므로 본 연구에서 제안하는 방법론은 기존의 SPRA 정량화 방법이 다룰 수 없는 시스템 내 부분 종속 조건을 고려하여 지진 리스크를 정확하게 평가할 수 있는 유용한 도구로 활용될 수 있을 것이라 기대한다.

• Korean Journal of Acupuncture
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• 제38권3호
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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.

• Journal of Ginseng Research
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• 제45권2호
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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.

• Journal of Periodontal and Implant Science
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• 제51권3호
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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.

• 한국산학기술학회논문지
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• 제22권1호
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• pp.257-266
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• 2021

최근 전 세계적으로 기후변화로 인해 설계빈도를 초과하는 강우가 발생함에 따라 도시 지역의 침수피해가 종종 발생한다. 도시 침수방어 대책으로는 구조적 대책과 비구조적 대책이 있다. 본 연구에서는 비구조적 대책 중에서 내배수 시스템의 운영에 초점을 두었다. 제안 방법인 내외수 연계 운영 방법은 상류 지점에 모니터링 지점을 선정하여 빗물펌프장에 유입되는 유입량을 예측하고, 이를 바탕으로 펌프를 운영하는 방법이다. 본 연구에서는 내외수 연계 운영 기술을 목감천 유역에 적용하였으며, 서울특별시에 큰 침수피해가 발생했던 기왕 강우인 2010년, 2011년 사상을 바탕으로 내외수 연계 운영 방법을 검증하였다. 2010년, 2011년 강우 사상을 적용 결과, 기존 운영 방법대비 침수 저감 효과는 각각 34.9 %, 54.4 % 만큼 감소한 것으로 확인되었다. 즉, 본 연구에서 제안한 내외수 연계 운영 방법은 유수지의 추가 저류공간을 확보함으로써 도시 유역 내에서 침수피해를 최소화했음을 확인하였다. 또한, 집중호우가 많이 발생하는 도시지역에서 침수를 예방하고, 시민의 생명과 재산을 지킬 수 있을 것으로 기대한다.