• 한국산학기술학회논문지
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• 제22권1호
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• pp.431-438
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• 2021

본 연구는 향후 보다 강화될 것으로 예측되어지는 USCG Phase II 형식승인시험을 대비하기 위해 SYBR Green I과 SYBR Gold의 염색 효율을 비교한 후 염색 효율이 높은 시약을 실제 선박평형수처리장치(electrolysis type, UV + electrolysis type)를 통과한 처리수에 적용해서 보았다. 시료의 부피가 0.5 mL ~ 2 mL, 염색 시약(Stock solution)을 100배 및 200배 희석한 조건에서 염색된 바이러스가 가장 선명하게 관찰되었다. SYBR Green I과 SYBR Gold의 염색효율은 장목한 해수조건의 실험구에서 유의한 차이를 보이지 않았지만, SYBR Gold의 노란색에 비해 SYBR Green I으로 염색된 시료에서 발현하는 녹색 형광이 보다 선명해서 관찰이 용이한 것으로 확인되었다. 선박평형수처리장치(electrolysis type, UV + electrolysis type)를 통과하지 않은 실험수 및 대조수에서의 해양 바이러스 현존량은 약 109~1010 VLP 100 mL-1 으로 확인된 반면, 처리수에서는 살아 있는 바이러스가 관찰되지 않았다. 실험수 결과를 보면, SYBR Green I은 해수, 기수, 담수 조건에서 효과적으로 염색이 되는 것으로 확인되었다. 다양한 선박평형수처리기술에 따른 추가적인 검증 및 염색 방법 개발이 필요하지만, SYBR Green I 염색법은 USCG Phase II 미국형식승인시험 바이러스 생산판별에 좋은 대안이 될 수 있을 것으로 판단된다.

• 구강회복응용과학지
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• 제19권2호
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• pp.87-96
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• 2003

The purposes of this study were to find out the optimum intensity of magnetic field where magnetism could promote the activity of osteoblast, and to discover the possibility of clinical application in the areas of dental implants and bone grafts by confirming the effect of clinically increasing bone formation. In this experiment, we used the Neodymium magnet, which had magnetic power six times as strong as the current ones and enabled the resistances against the demagnetization up to 20 to 50 times to be minimized with the size of 1mm in sight. In order to culture cells, a specially designed device was used. It was made to adjust the distance and accordingly to control the intensity of the magnetic field, by placing the cell culture plate in the center with a magnet of 1mm long and thick installed on the both ends. Using MC3T3-E1 cell, a kind of osteoblast-like cell, we cultured, for 24 hours, not only the test group which had been cultured under the magnetic fields with different intensity of 5, 10, 50, 100, 500, and 1000 Gauss, but also the control group excluding the influences of the magnetic field. After observing the cell's form and the density of the culture medium through an inverted microscope, we made a series of proceedings needed for the immunofluoroscence staining, such as fixation, normal serum reaction, primary antibody reaction, and secondary antibody reaction. And with a fluorescence microscope, we observed those-above and compared the frequency of expression of IFG-1 receptor. To make a Western immunoblotting analysis, the cells cultured under the same condition as the above had the procedure of the lysis buffer and the acrylamide gel electrophoresis was carried out. Protein transferred into the nitrocellulose membrane and tested on the primary and the secondary antibody reactions was observed and compared. The results were as follows: When observed through an inverted microscope, the nuclear divisions of the cells under the magnetic field of 10 Gauss were the most active, and the density of the cells could be observed the most enormously. As the result of an immunofluoroscence staining of IGF-1 receptor, the expression of IFG-1 was the most frequently observed under the magnetic field of 10 Gauss. On the other hand, few differences of consideration were made between the test group cultured under the magnetic fields of 5, 500, and 1000 Gauss and the control group. In respect of the expression of IFG-1 receptor, the test group cultured under the magnetic fields of 50 and 100 Gauss were higher than the control group, and lower than that cultured under the magnetic field of 10 Gauss.(p<0.05) According to the Western immunoblotting analysis, the band of IFG-1 receptor which had 85KDa of molecular weight was the darkest. Judging from the above-mentioned results, the growth factor receptor of an osteoblast cell which was an important criterion for the bone formation was increased in maximum under the magnetic field of 10 Gauss. Moreover it was observed that the optimum intensity of magnetic field in which magnetism made the activity of the osteoblast cell increase was about 10 Gauss.

• 한국식품영양과학회지
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• 제41권4호
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• pp.437-442
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• 2012

홍국쌀 추출물의 항산화 작용을 DPPH radical 및 hydroxyl radical 소거능, 간암 세포에서의 산화적 DNA 손상과 항산화 효소의 유전자 발현에 미치는 영향으로 분석하였다. 홍국쌀 추출물의 DPPH radical 소거능은 ethyl acetate 추출물, methanol 추출물, butanol 추출물 순이었으며, ethyl acetate 추출물의 DPPH radical 소거능은 0.2 mg/mL에서 85%, $IC_{50}$는 0.13 mg/mL로 나타났다. Ethyl acetate 추출물의 hydroxyl radical 소거능을 DCF fluorescence로 측정한 결과, cuvette에서는 2.5 ${\mu}g$/mL에서 44.2%, 5.0 ${\mu}g$/mL 74.1%, 10.0 ${\mu}g$/mL >100%로 나타났고, $HepG_2$ cell에서는 ethyl acetate 추출물로 전처리한 세포의 radical이 $H_2O_2$로만 처리한 세포에 비해 유의적으로 감소하였다. 홍국쌀 ethyl acetate로 전처리한 세포의 DNA 손상이 $H_2O_2$로만 처리한 세포에 비해 유의적으로 낮았으며, 추출물 대신 lovastatin을 처리한 세포는 DNA 손상이 증가하는 것으로 나타났다. 항산화 효소 유전자의 상대적 발현 정도를 측정한 결과, 산화 스트레스 없이 ethyl acetate 추출물로 전처리한 세포에서는 SOD와 GPx가 control에 비해 각각 3.25배, 2.67배 유의적으로 증가하였으며, 추출물로 전처리한 후 산화 스트레스에 노출시킨 세포에서는 CAT가 control에 비해 5 ${\mu}g$/mL에서 4.64배, 10 ${\mu}g$/mL에서 7.0배 유의적으로 증가하였다.

• 생물환경조절학회지
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• 제27권4호
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• pp.285-293
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• 2018

본 연구는 환경 친화적 방법인 브러싱을 이용한 기계적 자극의 영향을 받는 오이와 토마토 플러그 묘의 생육 억제 효과를 구명하기 위해 수행되었다. 오이(Cucumis sativus L. 'Joeunbaekdadagi')와 토마토(Solanum lycopersicum L. 'Mini Chal')를 2017년 10월 9일 상업용 혼합 상토가 충진된 40구 플러그 트레이($54{\times}27.5{\times}5cm$)에 파종하였다. 벤로형 유리온실의 재배환경은 $15-25^{\circ}C$의 재배 온도 범위와 $50{\pm}10%$의 상대습도를 유지하였다. 파종 15일후에, 오이와 토마토 묘에 무처리(대조구), $7.5mg{\cdot}L^{-1}$의 diniconazole을 처리하였다. 또한, 오이와 토마토의 brushing 처리는 2, 4, 또는 6시간 간격으로 각각 15일과 20일간 적용되었다. 1회씩 brushing 처리를 하였다. 오이와 토마토의 초장, 하배축, 절간장은 대조구에 비해 diniconazole 처리에서 억제되었다. 잎의 크기는 오이와 토마토 모두 감소하였지만, 반면에 엽록소 값은 diniconazole 처리에서 증가하였다. 그러나 오이의 경경은 2시간 brushing 간격 처리에서 가장 두꺼웠다. 지상부와 지하부의 생체중은 diniconazole 처리에서 유의적으로 낮았다. Brushing의 적용은 토마토 묘의 지상부와 지하부의 건물중, 충실도를 촉진시킴으로써 묘소질을 향상 시켰다. 토마토 묘의 엽록소 형광은 2시간 처리에서 급격히 감소하였으며, 이는 brushing 처리에 의한 기계적 스트레스를 나타낸다. 토마토 묘의 상대 생장률은 diniconazole 처리에서 유의적으로 낮았지만, 오이 묘는 모든 처리에서 유의적인 차이가 없었다. 결과적으로, 오이와 토마토 묘의 생육 억제는 생장조절제의 화학 물질에 의한 diniconazole 처리에서 가장 효과적이었다. 그러나 환경 친화적인 관점에서, 2시간의 brushing 간격 처리는 오이와 토마토 묘의 생장에서 화학적 방법을 대체할 수 있는 응용 가능성을 가지고 있다고 판단된다.

• Journal of Plant Biotechnology
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• 제5권3호
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• pp.143-148
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorhodamine123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MSP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to playa novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.

• 한국진공학회:학술대회논문집
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• 한국진공학회 2012년도 제43회 하계 정기 학술대회 초록집
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• pp.386-386
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• 2012

For white light emitting diode (LED) applications, it has been reported that Y3Al5O12:Ce3+ (YAG:Ce) in nano-sized phosphor performs better than it does in micro-sized particles. This is because nano-sized YAG:Ce can reduce internal light scattering when coated onto a blue LED surface. Recently, there have been many reports on the synthesis of nano-sized YAG particles using bottom-up method, such as co-precipitation method, sol-gel process, hydrothermal method, solvothermal method, and glycothermal method. However, there has been no report using top-down method. Top-down method has advantages than bottom-up method, such as large scale production and easy control of doping concentration and particle size. Therefore, in this study, nano-sized YAG:Ce phosphors were synthesized by a high energy beads milling process with varying beads size, milling time and milling steps. The beads milling process was performed by Laboratory Mill MINICER with ZrO2 beads. The phase identity and morphology of nano-sized YAG:Ce were characterized by X-ray powder diffraction (XRD) and field-emission scanning electron microscopy (FESEM), respectively. By controlling beads size, milling time and milling steps, we synthesized a size-tunable and uniform nano-sized YAG:Ce phosphors which average diameters were 100, 85 and 40 nm, respectively. After milling, there was no impurity and all of the peaks were in good agreement with YAG (JCPDS No. 33-0040). Luminescence and quantum efficiency (QE) of nano-sized YAG:Ce phosphors were measured by fluorescence spectrometer and QE measuring instrument, respectively. The synthesized YAG:Ce absorbed light efficiently in the visible region of 400-500 nm, and showed single broadband emission peaked at 550 nm with 50% of QE. As a result, by considering above results, high energy beads milling process could be a facile and reproducible synthesis method for nano-sized YAG:Ce phosphors.

• 한국응용곤충학회지
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• 제38권2호
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• pp.139-144
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• 1999

Autographa californica 핵다각체병 바이러스(AcNPV)의 다각체 단백질과 초록색 형광 단백질의 융합단백질의 특성을 분석하였다. 초록색 형광 단백질 유전자는 AcNPV의 완전한 다각체 단백질 유전자의 앞쪽과 뒤쪽에 융합하여 다각체 단백질 유전자의 프로모터 조절하에 도입하였다. 이렇게 작성된 재조합 바이러스를 각각 Ac-GFPPOL 또는 Ac-POLGFP이라고 명명하였다. 이들 재조합 바이러스에 의해 감염된 곤충세포주에서는 56kDa의 융합단백질이 발현되었다. 한편, 흥미롭게도 재조합 바이러스 Ac-POLGFP에 의해 감염된 세포주에서는 초록색 형광이 핵내에서만 다각체 유사 granular particle 형태로 관찰되었다. 반면에 Ac-GFPPOP에 의해 감염된 세포도주에서는 대부분 핵내에 존재하였지만, 세포질과 핵 모두에서 초록색 형광을 관찰할 수 있었다. 그러나 발현된 융합단백질은 분명히 다각체단백질을 포함하고 있음에도 다각체는 형성하지 않았다. 이러한 결과들은 융합단백질에서 다각체단백질의 위치와 관련이 있는 것으로 보여진다.

• Asian Pacific Journal of Cancer Prevention
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• 제13권11호
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• pp.5391-5397
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• 2012

Background: Neuroblastoma (NB), like most human cancers, is characterized by genomic instability, manifested at the chromosomal level as allelic gain, loss or rearrangement. Genetics methods, as well as conventional and molecular cytogenetics may provide valuable clues for the identification of target loci and successful search for major genes in neuroblastoma. We aimed to investigate AURKA and MYCN gene rearrangements and the chromosomal aberrations (CAs) to determine the prognosis of neuroblastoma. Methods: We performed cytogenetic analysis by G-banding in 25 cases [11 girls (44%) and 14 boys (66%)] and in 25 controls. Fluorescence in situ hybridization (FISH) with AURKA and MYCN gene probes was also used on interphase nuclei to screen for alterations. Results: Some 18.4% of patient cells exhibited CAs., with a significant difference between patient and control groups in the frequencies (P<0.0001). Some 72% of the cells had structural aberrations, and only 28% had numerical chnages in patients. Structural aberrations consisted of deletions, translocations, breaks and fragility in various chromosomes, 84% and 52% of the patients having deletions and translocations, respectively. Among these expressed CAs, there was a higher frequency at 1q21, 1q32, 2q21, 2q31, 2p24, 4q31, 9q11, 9q22, 13q14, 14q11.2, 14q24, and 15q22 in patients. 32% of the patients had chromosome breaks, most frequently in chromosomes 1, 2, 3, 4, 5, 8, 9, 11, 12, 19 and X. The number of cells with breaks and the genomic damage frequencies were higher in patients (p<0.001). Aneuploidies in chromosomes X, 22, 3, 17 and 18 were most frequently observed. Numerical chromosome abnormalities were distinctive in 10.7% of sex chromosomes. Fragile sites were observed in 16% of our patients. Conclusion: Our data confirmed that there is a close correlation between amplification of the two genes, amplification of MYCN possibly contributing significantly to the oncogenic properties of AURKA. The high frequencies of chromosomal aberrations and amplifications of AURKA and MYCN genes indicate prognostic value in children with neuroblastomas and may point to contributing factors in their development.

• 한국가축번식학회지
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• 제21권4호
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• pp.335-343
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• 1997

The effect of thiol compounds on development and intracellular glutathione(GSH) concentrations of bovine embryos produced by in vitro maturation and in vitro fertilization(IVM/IVF) was examined in CRlaa medium with or without $\beta$-mercaptoethanol(0, 10, 25 and 50$\mu$MME) and cysteamine(0, 25, 50 and 75 $\mu$M). Numbers of cells comprising blastocysts were also counted using double fluorescence stain and the total glutathione levels(oxidized and reduced form) of morula and blastocyst embryos were than measured by an enzymatic method. Following routine IVM/IVF procedures oocytes and zygotes were cultured for 40 to 44h in CRlaa medium. Then 2 to 8-cell embyos had cumulus cell removed and were allotted randomly to the experimental medium. In Experiment 1, the proportion of embryos developing to and beyond morulae stages in 0, 10, 25 and 50 $\mu$M $\beta$-ME was 42.9%, 50.0%, 53.7% and 65.6%, respectively. Fifty $\mu$M $\beta$-ME group was significantly higher than those of any other groups (P<0.05). In Experiment 2, the percentages of embryos developed beyond morulae stages in 0, 25, 50 and 75 $\mu$M cysteamine was 42.9%, 40.4%, 60.0% and 59.2%, respectively. Fifty and 75$\mu$M cysteamine groups were significantly higher than in 0 and 25 $\mu$M cysteamine groups, but all of culture medium containing cysteamine(52.6%) was not significantly difference in control group(42.9%). In Experiment 3, the intracellular GSH concentrations of morulae and blastocyst embryos in 0 and 50 $\mu$M $\beta$-ME was 42.4 pM and 44.9 pM, 49.5 pM and 67.8 pM, respectively. Morulae embryos were not difference, but blastocyst embryos were significantly difference between treatments(P<0.05). In Experiment 4, the intracellular GSH concentrations of morulae in CRlaa with or without cysteamine were 39.8 pM and 45.6 pM, and blastocysts were 59.3 pM and 66.8 pM, respectively. Cell numbers of blastocysts were similar to in all experimental groups. These experiments indicate that thiol compounds can increase the proportion of embryos that developing to and beyond morulae stage and the intracellular GSH concentrations.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2003년도 식물바이오벤처 페스티발
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• pp.65-71
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• 2003

Bax, a mammalian pro-apoptotic member of the Bcl-2 family, induces cell death when expressed in yeast. To investigate whether Bax expression can induce cell death in plant, we produced transgenic Arabidopsis plants that contained murine Bax cDNA under control of a glucocorticoid-inducible promoter. Transgenic plants treated with dexamethasone, a strong synthetic glucocorticoid, induced Bax accumulation and cell death, suggesting that some elements of cell death mechanism by Bax may be conserved among various organisms. Therefore, we developed novel yeast genetic system, and cloned several Plant Bax Inhibitors (PBIs). Here, we report the function of two PBIs in detail. PBI1 is ascorbate peroxidase (sAPX). Fluorescence method of dihydrorho-damine 123 oxidation revealed that expression of Bax in yeast cells generated reactive oxygen species (ROS), and which was greatly reduced by co-expression with sAPX. These results suggest that sAPX inhibits the generation of ROS by Bax, which in turn suppresses Baxinduced cell death in yeast. PBI2 encodes nucleoside diphosphate kinase (NDPK). ROS stress strongly induces the expression of the NDPK2 gene in Arabidopsis thaliana (AtNDPK2). Transgenic plants overexpressing AtNDPK2 have lower levels of ROS than wildtype plants. Mutants lacking AtNDPK2 had higher levels of ROS than wildtype. $H_2O_2$ treatment induced the phosphorylation of two endogenous proteins whose molecular weights suggested they are AtMPK3 and AtMPK6. In the absence of $H_2O_2$ treatment, phosphorylation of these proteins was slightly elevated in plants overexpressing AtNDPK2 but markedly decreased in the AtNDPK2 deletion mutant. Yeast two-hybrid and in vitro protein pull-down assays revealed that AtNDPK2 specifically interacts with AtMPK3 and AtMPK6. Furthermore, AtNDPK2 also enhances the MBP phosphorylation activity of AtMPK3 in vitro. Finally, constitutive overexpression of AtNDPK2 in Arabidopsis plants conferred an enhanced tolerance to multiple environmental stresses that elicit ROS accumulation in situ. Thus, AtNDPK2 appears to play a novel regulatory role in $H_2O_2$-mediated MAPK signaling in plants.