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Overexpression and Activity Analysis of Cystathionine γ-Lyase Responsible for the Biogenesis of H2S Neurotransmitter (새로운 신경전달물질 H2S 발생 효소, cystathionine γ-lyase의 대량발현 조건과 활성측정)

  • Kim, Kyoung-Ran;Byun, Hae-Jung;Cho, Hyun-Nam;Kim, Jung-Hyun;Yang, Seun-Ah;Jhee, Kwang-Hwan
    • Journal of Life Science
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    • v.21 no.1
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    • pp.119-126
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    • 2011
  • There is a growing recognition of the significance of $H_2S$ as a biological signaling molecule involved in vascular and nervous system functions. In mammals, two enzymes in the transsulfuration pathway, cystathionine ${\beta}$-synthase (CBS) and cystathionine ${\gamma}$-lyase (CGL), are believed to be chiefly responsible for $H_2S$ biogenesis. Genetic inborn error of CGL leads to human genetic disease, cystathioninuria, by accumulating cystathionine in the body. This disease is secondarily associated with a wide range of diseases including diabetes insipidus and Down's syndrome. Although the human CGL (hCGL) overexpression is essential for the investigation of its function, structure, reaction specificity, substrate specificity, and protein-protein interactions, there is no clear report concerning optimum overexpression conditions. In this study, we report a detailed analysis of the overexpression conditions of the hCGL using a bacterial system. Maximum overexpression was obtained in conditions of low culture temperature after inducer addition, performing low aeration during overexpression, and using a low concentration inducer (0.1 mM, IPTG) for induction. Expressed hCGL was purified by His-tag affinity column chromatography and confirmed by Western blot using hCGL antibody and enzyme activity analysis. We also report that the His tag with TEV site attached protein exhibits 76% activity for ${\alpha}-{\gamma}$ elimination reaction with L-cystathionine and 88% for ${\alpha}-{\beta}$ elimination reaction with L-cysteine compared to those of wild type hCGL, respectively. His tag with TEV site attached protein also exhibits a 420 nm absorption maximum, which is attributed to the binding cofactor, pyridoxal 5'-phosphate (PLP).

Analysis of Putative Downstream Genes of Arabidopsis AtERF71/HRE2 Transcription Factor using a Microarray (마이크로어레이를 이용한 애기장대 AtERF71/HRE2 전사인자의 하위 유전자 분석)

  • Seok, Hye-Yeon;Lee, Sun-Young;Woo, Dong-Hyuk;Park, Hee-Yeon;Moon, Yong-Hwan
    • Journal of Life Science
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    • v.22 no.10
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    • pp.1359-1370
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    • 2012
  • Arabidopsis AtERF71/HRE2, a transcription activator, is located in the nucleus and is involved in the signal transduction of low oxygen and osmotic stresses. In this study, microarray analysis using AtERF71/HRE2-overexpressing transgenic plants was performed to identify genes downstream of AtERF71/HRE2. A total of 161 different genes as well as AtERF71/HRE2 showed more than a twofold higher expression in AtERF71/HRE2-overexpressing transgenic plants compared with wild-type plants. Among the 161 genes, 24 genes were transcriptional regulators, such as transcription factors and DNA-binding proteins, based on gene ontology annotations, suggesting that AtERF71/HRE2 is an upstream transcription factor that regulates the activities of various downstream genes via these transcription regulators. RT-PCR analysis of 15 genes selected out of the 161 genes showed higher expression in AtERF71/HRE2-overexpressing transgenic plants, validating the microarray data. On the basis of Genevestigator database analysis, 51 genes among the 161 genes were highly expressed under low oxygen and/or osmotic stresses. RT-PCR analysis showed that the expression levels of three genes among the selected 15 genes increased under low oxygen stress and another three genes increased under high salt stress, suggesting that these genes might be downstream genes of AtERF71/HRE2 in low oxygen or high salt stress signal transduction. Microarray analysis results indicated that AtERF71/HRE2 might also be involved in the responses to other abiotic stresses and also in the regulation of plant developmental processes.

Study on Korean Ancient Dietary Culture through Japanese Sacrificial Offerings -Danjanjinja Kakitsisai(2)- (일본신찬(日本神饌)을 통한 한국고대식(韓國古代食)의 추정연구(推定硏究) -담산신사(談山神社) 가길제(嘉吉祭) 백미어식(百味御食)(2)-)

  • Kim, Chon-Ho;Kaneko, Kentaro;Sumino, Takeshi;Kaneda, Takashi
    • Journal of the Korean Society of Food Culture
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    • v.8 no.2
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    • pp.139-146
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    • 1993
  • The predeccessor of Danjanjinja was Myorak temple which is built in the 7th century. At that age, the Buddhist culture of Japan had highly prospected by transmitting Buddhism to Japan from Han peninsular On the other hand, the private god of Fujiwara family in Danjanjinja is Uchigami which is one of typical Japanese popular belief like Dangshin of Korean's. Through these historical background, it could by presumed that the Kakitsisai might be the original form of Korean Buddhist sacrificial offerings from ancient age. So this study on Kakitsisai what had handed down from generation to generation about for 1300 years help us to study and estimate the ancient dietary culture of Korean and Japanese. 1. Kakitsisai performed high filling method in the sacrificial offerings like Kasuga, Horyuji and Korea. 2. The patterns and colors of high filling offerings are various in Korea and Japan. 3. They used unpolished rice by ancient rice, and called red and black one. We can guess both of countries ate unpolished rice at that age. 4. They used many kind of ancient wild fruits and vegetables. We could recognize what the ancients had eaten the foods.

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Detection of Lamivudine-Resistant Mutations of HBV DNA Polymerase Gene Using PCR-Direct Sequencing

  • Lee, Kyung-Ok;Lee, Hye-Jung;Byun, Ji-Young;Lee, Sung-Yeun;Kim, Jeong-Sook;Jung, Na-Young;Chung, Soo-Jin;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.196-202
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    • 2006
  • Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

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Oxygen Toxicity of Superoxide Dismutase-Deficient Saccharomyces cerevisiae by Paraquat (Paraquat에 의해 유도된 Superoxide Dismutase 결핍 효모의 산소 독성)

  • 김지면;남두현용철순허근
    • KSBB Journal
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    • v.10 no.5
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    • pp.561-567
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    • 1995
  • Using superoxide dismutase (SOD)-deficient mutants of Saccharomyces cerevisiae, the oxygen toxicity induced by paraquat was studied. In aerobic culture condition, yeasts lacking MnSOD (milochondrial SOD) showed more significant growth retardation than CuZnSOD (cytoplasmic SOD)-deficient yeasts. However, not so big differences in growth pattern of those mutants compared with wild type were observed under anaerobic condition. When exposed to paraquat, the growth of yeasts lacking CuZnSOD was severely affected by higher than 0.01mM of paraquat in culture medium. By the analysis of several cellular components ivolved in free radical generating and scavenging system, it was found that, under aerobic condition, the content of lipid peroxides in cell membrane as well as cellular activity of glutathion peroxidase of CuZnSOD-deficient mutants was increased in the presence of paraquat, although significant decrease of catalase activity was observed in those stratns. In MnSOD-deficient yeast, however, increment in cellular activity of glutathion peroxldase and catalase by paraquat was observed without any deterioration of membrane lipid. It implies that the lack of mitochondrial SOD could be compensated by both of glutathion peroxldase and catalase, but that only glutathion peroxidase might act for CuZnSOD in cytoplasm. In contrast, all of SOD-deficient mutants showed a significant decrease in catalase activity, but slight increase in the activities of glutathion peroxidase, when cultivated anaerobically in the medium containing paraquat. Nevertheless, any significant changes of lipid peroxides in cell membranes were not observed during anaerobic cultivation of SOD-deficient mutants. It suggests that a little amount of free radicals generated by paraquat under anaerobic condition could be sufficiently overcome by glutathion peroxidase but not by catalase.

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The Growth and Physiological Responses of Cacalia firma Seedlings by Shading Conditions in Forest Farming (임간재배 시 병풍쌈 유묘의 차광처리별 생장 및 생리 반응)

  • Yoon, Jun Hyuck;Jeon, Kwon Seok;Song, Ki Seon;Park, Yong Bae;Moon, Yong Sun;Lee, Do Hyung
    • Journal of Korean Society of Forest Science
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    • v.103 no.1
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    • pp.65-71
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    • 2014
  • Cacalia firma is a perennial plant in Asteraceae, Parasenecio that distributed in Korea, China, and Japan. As dietary style changes for well-being life, consumer's demand of functional food and organic vegetables is getting increased. This study was conducted to investigate the optimum light conditions of P. firmus in forest farming. One year old seedlings were grown under four different light conditions 10%, 20%, 30%, and 50% of sunlight by shading (equals 50%, 30%, 20%, and 10% relative brightness respectively) and non-treated control under full sunlight. They were analyzed for early growth and physiological response. Seedlings grown under 75% shading showed similar height, root growth, and leaf water content to control. However, their leaf length, width, and total leaf area were increased, which caused increased leaf dry weight and total dry weight. Especially, seedlings under 95% shading showed 40% increase in height and more leaf growth and leaf water content, although they had shorter main root length and root collar diameter than control. In addition specific leaf area (SLA) and leaf area ratio (LAR) were higher than control and indicated that they were statistically significant difference from control. Higher SLA refers thinner leaf thickness, higher LAR means larger leaf area. The results indicate seedlings under 95% shading have higher water content, thinner leaf, and wider lightinterception areas. It is plausible that P. firmus is active in chlorophyll activities and carbon dioxide assimilation at even lower light conditions. These results suggest that the optimum light level of P. firmus for artificial cultivation in forest farming ranges from 75~95% shading (20%-10% of relative brightness). When salability as 'sanchae' (wild edible greens) is considered, P. firmus could be cultivated under 75% shading in forest farming and expected to have better taste and higher yield. We suggest these results as basic data of P. firmus for possible forest farming.

Converting Lands that are damaged by Graveyards into Tree Burial Sites in order to Restore Green Areas (산지묘지의 훼손지 복원을 위한 수목장지로의 전환)

  • Woo, Jae-Wook;Byun, Woo-Hyuk;Kim, Hak-Beom;Park, Won-Kyoung;Kim, Min-Su;Norsyuhada, Norsyuhada
    • Journal of the Korean Institute of Landscape Architecture
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    • v.40 no.3
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    • pp.69-80
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    • 2012
  • The purpose of this paper was to study the issues related to converting the graveyards within forests into spaces intended for tree burials by means of planting, given the situation that the graveyards have encroached on land and damaged the environment. For the reason, a field survey was performed to determine the width, length, and distance to the nearest tree of 205 graveyards in the capital area. Through this, it was determined that the domestic lands damaged by graveyards amounted to $862km^2$, including the areas that were deforested to manage the graves. This only confirms that land encroachment by graveyards is a serious issue. The methods for making tree burial sites were examined from the perspective of how to meet public demands given the graveyard's spatial distinctiveness. As a result, this study suggested different methods to establish tree burial sites according to the degree of transformation and the term of its formation. This study also classified the graveyards into three types, and identified the planting methods that harmonized the safe growth of trees and the scenic beauty of memorial places based on the standard. This is in order to plant trees that are shade-tolerant and suitable to the forest line, along with which other tree line was and also, to plant aesthetic trees around the empty space. Through applying the developed methods, this study established and monitored two exemplary sites in Yongin and Boryeng. Aesthetic trees were planted in Yongin site which was located in an open area, aod the shade-tolerant trees were planted in Boryeong, which was located in a forest area. As a result, the image of a garden appeared at Yongin site and the image of a tree colony harmonized with the near forest emerged at Boryeong site. Therefore, it is confirmed that the method of planting according to the distribution status of neighboring trees was effective. As a result of monitoring, mulching wood chips were suitable for sites that were small or easy to approach. This is because the weeds were controlled in Yongin site by mulching. Furthermore, by monitoring the growth of 11 species of vegetation, this study confirmed that low and cover-type vegetations were suitable for tree burial sites. In Boryeong site, the wild cherry trees, which were planted as adult trees, all died, and the tilling of snake's beard, which were planted as cover vegetation, was slow. Therefore, this study found that seedlings were more suitable to plant in forest graveyards than adult trees, which were large and difficult to approach, and it was effective to use the remaining lawn and form a low vegetation after the crown of trees had expanded to such places.

An Ecological Aesthetics and Symbolism of the Seonghyelsa Nahanjeon Floral Lattice with Patterns of Lotus Pond Scenery (연지(蓮池)로 본 성혈사 나한전 꽃살문양의 생태미학과 상징성)

  • Rho, Jae-Hyun;Lee, Da-Young;Choi, Seung-Hee
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.36 no.3
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    • pp.160-171
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    • 2018
  • This study aims to find an original form of temple flower decoration patterns, considering floral lattice pattern as a view element composing temple landscape. To that end, we analyzed and interpreted the form and symbol expressed in the floral lattice pattern at Nahanjeon of Seonghyel Temple at Yeongju, Gyeongsangbukdo. The front side of Nahanjeon windows shows a sculpture with 176 pure patterns in a form where two squares are in sequence. The basic concept of main front door (the inner gate of Nahanjeon) frames is considered the design language of lotus pond that symbolizes "square land" in traditional gardens. The four leaf clover and arrowhead are water plants discovered in areas nearby ponds, which are a realistic expression conforming to the water ecology of lotus pond. The lotus, which is the most important plant at the main front door, indicates purity, a non-stained state, and the world of the lotus sanctuary, which is the land of blissful happiness in Buddhism. The lotus expressed in the floral lattice pattern is spread in a diverse form, containing the features of creation and destruction, showing the landscape character of the "One Body of Buddha and Lotus". The expression of flying birds such as kingfishers and egrets is an ecologically aesthetic idea to infuse dynamism and vitality into a seemingly static aquatic ecosystem. The floral lattice pattern contains lotus pond scenery showing symbiosis of animals(i.e., dragons, frogs, crabs, fishes, egrets, wild geese, and kingfishers) and plants(i.e., four leaf clovers and arrowheads), which are symbols of relief faith for longevity, wealth, preciousness, and many sons. The pattern is not just an ecological aesthetic expression but a holistic harmony of ecological components such as growth and disappearance of lotus and its leaves, fitting habitats, symbiosis, and food chain.

Comparative Evaluation on Sperm Parameter of Transgenic Pigs with General Pigs (형질전환 돼지의 정자와 일반돼지의 정자성상에 대한 비교평가)

  • Park, Sang Hyoun;Lee, Gunsup;Lee, Joo Yung;Kim, Kyung Woon;Byun, Sung-June;Ock, Sun A;Hwang, Seongsoo;Yang, Hyeon;Woo, Jae-Seok;Oh, Keon Bong
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.227-233
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    • 2017
  • Pig has been known to be one of the most feasible animals as a bioreactor to produce pharmaceuticals in milk and as a mediator in xenotransplantation research. Previously, we generated transgenic pigs for both purposes, which were expressing Factor 8, vWF, hTPA, and hEPO in milk, along with expression of MCP at GalT gene locus ($GalT^{-MCP/-MCP}$) as well as expressing MCP at GalT gene loci with CD73 expression ($GalT^{-MCP/+}/CD73$). In this study, we performed comparative analyses of sperm parameters between wild type male (WT) pig and those transgenic males to examine the effects of transgenes integrated into the pigs on motility, morphology, viability, and acrosome integrity of the spermatozoa. Our results showed that the rates of actively motile spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 85.0%, 83.3%, 82.5%, 83.3%, 82.5%, 77.5%, and 78.7%, respectively. Whereas, the rates of morphologically normal spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 90.0%, 80.0%, 80.0%, 83.3%, 85.0%, 91.8%, and 80.8%, respectively. In addition, the viability in spermatozoa of WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 93.9%, 82.4%, 89.9%, 83.9%, 87.4%, 92.8%, and 83.6%, respectively. The rates of spermatozoa with normal acrosome integrity in WT, Factor 8, vWF, hTPA, hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs were 98.1%, 98.6%, 98.6%, 98.7%, 98.1%, 99.5%, and 95.1%, respectively. There were no significant differences in motility, morphology, viability, and acrosome integrity of the spermatozoa among WT, Factor 8, vWF, hTPA, and hEPO, $GalT^{-MCP/+}/CD73$, and $GalT^{-MCP/-MCP}$ pigs. These mean that neither random integration nor targeted integration of the transgene into chromosome of pig effect on characteristics of spermatozoa. Ultimately, the transgenic male pigs subjected in this study could apply to propagate their progenies for production of human therapeutic proteins and advancing the xenotransplantation research.

A Study on Facilities Damage Characteristics Caused by Forest Fire in Goseong-Gun (고성산불로 인한 시설물피해특성 연구)

  • Yeom, Chanho;Lee, Si-young;Park, Houngsek;Kwon, Chungeun
    • Journal of the Society of Disaster Information
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    • v.15 no.4
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    • pp.469-478
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    • 2019
  • Purpose: In this studies we examine the facilities damage characteristics caused by forest fire. Therefore, we surveyed damaged facilities from forest fire which was occurred on Goseong-Gun on march 28 in 2019.(damaged areas was 40ha) The types of facilities uses were house, public facility, warehouse and so on. 17 facilities were destroyed. The purpose of this study was to for establishing a disaster safety village in rural areas where damage from a similar type of disaster occurs repeatedly by conducting the consciousness survey targeting at experts and disaster safety officials in a local government. Method: We surveyed meteorological factors(temperature, wind speed, wind direction, humidity) per a minute for analyzing weather condition on Goseong-Gun when forest fire was occurred, spread and extinguished. And we surveyed forest fire risk factors(a slope degree, a slope direction, a geographical feature, a distance between forest and facility, main species, the existence of crown fire ignition, the direction of facility, the main material of building) around 10 damaged facilities. Finally, we analyzed damage pattern of facilities using meteorological factor and forest fire reisk fator Result: The weather condition of Kanseonng AWS (No.517) was high temperature, arid and strong wind, when the forest fire was occurred and spread. An average wind speed was 4.1m/s and the maximum wind speed was 11.6m/s. The main direction of wind was W(225~315°). Damaged facilities were located on the steep slope area and on the mountaintop. The forest density around facilities was high and main species was korean red pine. The crown fire was occurred in the forest around damaged facilities. The average distance was 13.5m from forest to facilities. When the main matarial of building was made by fire resistance materials (for example, rainforced concrete), the damage was slightly. on the other hand, when by flammable material (for example, a Sandwich Panel), the facilities were totally destroyed Conclusion: The results of this research which were the thinning around house, making a safety distance, the improvement of main material of building and etc, will be helpful for establishing a counter measure for a forest fire prevention of facilities in wild land urban interface