• Title/Summary/Keyword: Isolated System

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Genetic Characterization for Human Enterovirus Isolated from Busan Region in Korea (부산지역에서 분리된 엔테로바이러스의 유행양상 분석 및 유전자형 연구)

  • Kim, Nam-Ho;Min, Sang-Kee;Park, Eun-Hee;Park, Yon-Koung;Kwan, Sun-Mok;Jin, Seong-Hyeon
    • Journal of Life Science
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    • v.20 no.6
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    • pp.907-913
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    • 2010
  • Human enteroviruses (HEV) are considered one of the major infectious causes of central nervous system infections such as aseptic encephalomeningitis in pediatrics. This study was focused on providing information related to genetic characteristics and diversities of HEV which prevailed between 2007 and 2009 in Busan, Korea. A total of 2,743 specimens were collected from children and screened for isolation of HEV by cell culture and RT-PCR. Among the specimins, 240 isolates were grouped into 21 different HEV serotypes using VP1 RT-PCR. The major etiological agents were CV-A6 and CV-B2 in 2007, E-6 and E-30 in 2008 and CV-B1 in 2009. The occurrence of HEV infections was the most frequent in the summer (May to August, 188 cases, 78.3%). Most of the isolates were identified from specimens from children under 10 years old, with the highest occurrence in the 2 to 4 year old range (15.2%). However, there were no significant differences between male and female children for the isolates. For analyzing genetic characterization, VP1 gene was amplified by RT-PCR and sequenced. The phylogenetic tree was established by Clustal W method using DNASTAR software. Using the sequence analysis of the VP1 region, it was classified into 2 groups; HEV-A and HEV-B. The HEV-A group contained 6 serotypes and sequences of 31 isolates were compared within each serotype. The HEV-B group contained 10 serotypes and the sequences of 41 isolates were compared within each serotype. Homology analysis of the VP1 region showed that the identity scores of HEV-A and B isolates were different. In conclusion, genetic divergences were observed among the isolates from children between 2007 and 2009 in Busan.

Analysis of Quorum Sensing-Related Phenotypes of Pseudomonas aeruginosa Clinical Isolates (녹농균 임상균주의 쿼럼 센싱 관련 표현형 분석)

  • Jung, Kyung-Ju;Choi, Yu-Sang;Ha, Chang-Wan;Shin, Jeong-Hwan;Lee, Joon-Hee
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.240-247
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    • 2010
  • Pseudomonas aeruginosa is a Gram (-) opportunistic human pathogen causing a wide variety of infections on lung, urinary tract, eyes, and burn wound sites and quorum sensing (QS), a cell density-sensing mechanism plays an essential role in Pseudomonas pathogenesis. In order to investigate the importance of QS in the Pseudomonas infections of Korean patients, we isolated 189 clinical strains of P. aeruginosa from the patients in Pusan Paik Hospital, Busan, South Korea. The QS signal production of these clinical isolates was measured by signal diffusion assay on solid media using reporter strains. While most clinical strains (79.4%) produced the QS signals as similar level as a wild type strain, PAO1 did, where LasR, the initial QS signal sensor-regulator was fully activated, a minority of them (4.2%) produced much less QS signals at the level to which LasR failed to respond. Similarly, while 72.5% of the clinical isolates produced QS signals enough to activate QscR, an another QS signal sensor-regulator, some few of them (9%) produced the QS signals at much lower level where QscR was not activated. For further analysis, we selected 74 clinical strains that were obtained from the patients under suspicion of Pseudomonas infection and investigated the total protease activity that is considered important for virulence. Interestingly, significant portion of them showed very low protease activity (44.6%) or no detectable protease activity (12.2%). When the biofilm-forming ability that is considered very important in chronic infection was examined, most isolates showed lower biofilm-forming activity than PAO1. Similarly, significant portion of clinical isolates showed reduced motility (reduced swarming activity in 51.4% and reduced twitching activity in 41.9%), or non-detectable motility (swarming-negative in 28.4% and twitching-negative in 28.4%). Our result showed that the clinical isolates that produced QS signals at the similar level to wild type could have significantly reduced activities in the protease production, biofilm formation, and motility, and some clinical isolates had unique patterns of motility, biofilm formation, and protease production that are not correlated to their QS activity.

A Study on Bone Formation & Osteoporosis by Taeyoungion-Jahage Extracts

  • Kim, Yi-Geun;Seong, Jun-Ho;Kim, Dong-Il;Lee, Tae-Kyun;Kim, Jun-Ki;Park, Young-Duck
    • The Journal of Korean Obstetrics and Gynecology
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    • v.15 no.4
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    • pp.45-60
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    • 2002
  • Mouse calvarial osteoblast cells were isolated and cultured. To examine whether the cells produce active gelatinases in culture medium or not,the cells were analyzed using by zymograsphic analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). We show that mouse calvarial osteoblasts in culture constitutively synthesize progelatinase- A. Then, mouse osteoblasts, which were stimulated by PTH, $1,25(OH)_2D_3$, mononuclear cell conditioned medium (MCM) and IL-1 as bone resorption agent's, showed increased collagenolysis by producing the active gelatinase. However, treatment of indomethacin and dexamethasone significantly decreased those effects of collagenolysis in mouse osteoblastic cells. On the other hand, IL-1 in stimulating bone resorption was examined using fetal mouse long bone organ culture. IL-1 stimulated bone resorption and produced marked resorption when present simultaneously. Furthermore, when it was examined the effects of indomethacin and dexamethasone on the dose dependent responses of $IL-1{\alpha}$, indomethacin and dexametasone produced a rightward shift in the IL-1 dose response curve. The results of in vitro cytotoxicities showed that Taeyoungjon-Jahage water extracts(T.Y.J-J.H.G extracts) have no any cytotoxicities in concentrations of $1-200\;{\mu}g/ml$ and furthermore there is no any cytotoxicity even in concentration of $300\;{\mu}g/ml$ on mouse calvarial bone cells. T.Y.J.-J.H.G. extracts had protective activity against PTH (2 units/mI), or MCM (5%, v/v), or $rhIL-1{\alpha}$ (1 ng/mI) or $1,25(OH)_2D3$ (10 ng/ml) , $IL-1{\alpha}$ and $IL-1{\beta}-induced$ collagenolysis in the mouse calvarial cells. Pretreatment of the T.Y.J.-J.H.G.extracts for 1 h, which by itself had little effect on cell survival, did not enhance the collagenolysis, nor significantly reduced the collagenolysis by pretreatment. Furthermore. the medicinal extracts were shown to have the protective effects against collagenolysis induced by $IL-1{\alpha}$ and $IL-1{\beta}$. Pretreatment of the extracts for 1 h significantly reduced the collagenolysis. Interestingly, the T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against gelatinase enzyme and processing activity induced by the bone resortion agents of PTH, $1,25(OH)_2D_3$, $IL-1{\beta}$ and $IL-1{\alpha}$, with strong protective effect in pretreatment with the extracts. T.Y.J.-J.H.G. extracts were shown to have the inhibiting effects against $IL-1{\alpha}-$ and $IL-1{\beta}-stimulated$ bone resorption and the effect of the pretreatment with a various concentrations of the medicinal extracts were significant. The inhibition extent and phenomena of IL-1 stimulated bone resorption by nonsteroidal anti-inflammatory agents of indomethacin and dexamethasone were similar to those obtained by T.Y.J.-J.H.G. extracts treatment in the mouse calvarial tissue culture system. These results indicated that the T.Y.J.-J.H.G.-water extracts are highly stable and applicable to clinical uses in osteoporosis.

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High Yield Bacterial Expression and Purification of Active Cytochrome P450 p-coumarate-3-hydroxylase (C3H), the Arabidopsis Membrane Protein (대장균 시스템을 이용한 Arabidopsis 막 단백질 cytochrome P450 p-coumarate-3hydroxylase (C3H) 활성형의 과발현 및 분리정제)

  • Yang, Hee-Jung;Kim, Wan-Yeon;Yun, Young-Ju;Yoon, Ji-Won;Kwon, Tae-Woo;Youn, Hye-Sook;Youn, Bu-Hyun
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1039-1046
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    • 2009
  • The cytochrome P450s (P450s) metabolizing natural products are among the most versatile biological catalysts known in plants, but knowledge of the structural basis for their broad substrate specificity has been limited. The activity of p-coumarate 3-hydroxylase (C3H) is thought to be essential for the biosynthesis of lignin and many other phenylpropanoid pathway products in plants however, all attempts to express and purify the protein corresponding C3H gene have failed. As a result, no conditions suitable for the unambiguous assay of the enzyme are known. The detailed understanding of the mechanism and substrate-specificity of C3Hdemands a method for the production of active protein on the milligram scale. We have developed a bacterial expression and purification system for the plant C3H, which allows for the quick expression and purification of active wild-type C3H via introduction of combinational mutagenesis. The modified cytochrome P450 C3H ($C3H_{mod}$) could be purified in the absence of detergent using immobilized metal affinity chromatography and size exclusion chromatography following extraction from isolated membranes in a high salt buffer and catalytically activated. This method makes the use of isotopic labeling of C3H for NMRstudies and X-ray crystallography practical, and is also applicable to other plant cytochrome P450 proteins.

BIOMECHANICS OF ABUTMENTS SUPPORTING REMOVABLE PARTIAL DENTURES UNDER UNILATERAL LOADING

  • Kim, Seong-Kyun;Heo, Seong-Joo;Koak, Jai-Young;Lee, Jeong-Taek;Roh, Hyun-Ki;Kim, Hyo-Jin;Lee, Seok-Hyung;Lee, Joo-Hee
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.6
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    • pp.753-759
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    • 2007
  • Statement of problem. In distal extension removable partial denture, the preservation of health of abutment teeth is very important, but abutment teeth are subjected to unfavorable stress under unilateral loading specially. Purpose. The purpose of this study was to investigate the biomechanical effects of mandibular removable partial dentures with various prosthetic designs under unilateral loading, using strain gauge analysis. Material and methods. Artificial teeth of both canines were anchored bilaterally in a mandibular edentulous model made of resin. Bilateral distal extension removable partial dentures with splinted and unsplinted abutments were fabricated. Group 1: Clasp-retained mandibular removable partial denture with unsplinted abutments Group 2: Clasp-retained mandibular removable partial denture with splinted abutments by 6-unit bridge. Group 3: Bar-retained mandibular removable partial denture Strain gauges were bonded on the labial plate of the mandibular resin model, approximately 2 mm dose to the abutments. Two unilateral vertical experimental loadings (30N and 100N) were applied subsequently via miniature load cell that were placed at mandibular left first molar region. Strain measurements were performed and simultaneously monitored from a computer connected to data acquisition system. For within-group evaluations, t-test was used to compare the strain values and for between-group comparisons, a one-way analysis of variance (ANOVA) was used and Tukey test was used as post hoc comparisons. Results. The strain values of group 1 and 2 were tensile under loadings. In contrast, strain values of group 3 were compressive in nature. Strain values increased as the applied load in increased from 30N to 100N (p<.05) except for right side in group 1. Under 30N loading, in left side, group 1 showed higher strain values than groups 2 and 3 in absolute quantity (p<.05). And group 2 showed higher strain values than group 1 (p<.05). In right side, group 1 and 2 showed higher strain values than group 3 in absolute quantity (p<.05). Under 100N loading in left side, group 1 showed higher strain values than groups 2 and 3 in absolute quantity (p<.05). And group 2 showed higher strain values than group 1 (p<.05). In right side, group 1 and 2 showed higher strain values than group 3 in absolute quantity (p<.05). Under 30N loading, group 2 and 3 showed higher strain values in right side than in left side. Under 100N loading, right side strain values were higher than left side ones for all groups. Conclusion. Splinting of two isolated abutments by bridge reduced the peri-abutment strain in comparison with unsplinted abutments under unilateral loading. Bar-retained removable partial denture showed the lowest strain of three groups, and compressive nature.

On the Utilization of Inactive BHC isomers -Synthesis of 3-(2,4,5-trichlorophenyl)-1-methyl urea as a herbicide- (BHC 이성질체(異性質體)의 활용(活用)에 관(關)한 연구(硏究) -제초제(除草劑)로서 3-(2,4,5-trichlorophenyl)-1- methyl urea의 합성(合成)-)

  • Lee, Kyu-Seung;Park, Chang-Kyu
    • Applied Biological Chemistry
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    • v.22 no.2
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    • pp.109-122
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    • 1979
  • Present study was carried out to reduce residual toxicity of BHC insecticides inherent in the organochlorine pesticides. For This end, r-isomer, the most potent insecticidal component among the BHC stereoisomers, was isolated and thus fortified by means of solvent precipitation. In parallel, 3-(2,4,5-trichlorophenyl)-1-methyl urea was prepared in good yield from technical BHC via 1,2,4-trichlorobenzene, 1,2,4,-trichloronitrobenzene, and 2,4,5-trichloroaniline. In addition, certain merit of the compound which make it possible to use as a herbicide is discussed. The results are summarized as follows; 1. Recrystallizing technical BHC from methanol-water binary solvent system, r-isomer was enriched to 49.7% at 95% recovery of r-isomer. 2. By partitioning technical BHC in 85% of methanolic solution into chloroform, r-isomer was fortified to 89.6% at 90.5% recovery of r-isomer. 3. Yield of 1,2,4-trichlorobenzene from technical BHC was greatly dependent upon concentration of alkalies and to less degree on the alkalies. 4. Surfactants, in particular cationic a quartenary ammonium salt, increased yield of 1,2,4-trichlorobenzene from technical BHC by alkaline hydrolysis. 5. Conversion of 1,2,4-trichlorobenzene to 2,4,5-trichloronitrobenzene was effected almost quantitatively utilizing $HNO_3-H_2SO_4$ nitrating agent at low temperature. 6. Yield of 91.4% was observed for the synthesis of 2,4,5-trichloroaniline by reducing 2,4,5-trichloronitrobenzene in the presence of iron turning and hydrochloric acid. 7. Overall yield based on BHC of 3-(2,4,5-trichlorophenyl)-1- methyl urea was 60.8%. 8. Inhibition effects, both germination and growth, 3-(2,4,5-trichlorophenyl)-1-methyl urea on several crops were found comparable to or more potent than those of $linuron{\circledR}\;and\;diuron{\circledR}$. In addition, it was also noted that susceptibility to the prepared compound depended upon the crops as well as specific part (shoots, roots) of the plant exposed to the chemicals.

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Development of Efficient Screening Methods for Melon Plants Resistant to Fusarium oxysporum f. sp. melonis (멜론 덩굴쪼김병에 대한 효율적인 저항성 검정법 개발)

  • Lee, Won Jeong;Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Heung Tae;Choi, Gyung Ja
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.70-82
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    • 2015
  • This study was conducted to establish an efficient screening system to identify melon resistant to Fusarium oxysporum f. sp. melonis. F. oyxsporum f. sp. melonis GR was isolated from infected melon plants collected at Goryeong and identified as F. oxysporum f. sp. melonis based on morphological characteristics, molecular analyses, and host-specificity tests on cucurbits including melon, oriental melon, cucumber, and watermelon. In addition, the GR isolate was determined as race 1 based on resistance responses of melon differentials to the fungus. To select optimized medium for mass production of inoculum of F. oxysporum f. sp. melonis GR, six media were tested. The fungus produced the most spores (microconidia) in V8-juice broth. Resistance degrees to the GR isolate of 22 commercial melon cultivars and 6 rootstocks for melon plants were investigated. All tested rootstocks showed no symptoms of Fusarium wilt. Among the tested melon cultivars, only three cultivars were susceptible and the other cultivars displayed moderate to high resistance to the GR isolate. For further study, six melon cultivars (Redqueen, Summercool, Superseji, Asiapapaya, Eolukpapaya, and Asiahwanggeum) showing different degrees of resistance to the fungus were selected. The development of Fusarium wilt on the cultivars was tested according to several conditions such as plant growth stage, root wounding, dipping period of roots in spore suspension, inoculum concentration, and incubation temperature to develop the disease. On the basis of the test results, we suggest that an efficient screening method for melon plants resistant to F. oxysporum f. sp. melonis is to remove soil from roots of seven-day-old melon seedlings, to dip the seedlings without cutting in s pore s uspension of $3{\times}10^5conidia/mL$ for 30 min, to transplant the inoculated seedlings to plastic pots with horticulture nursery media, and then to cultivate the plants in a growth room at 25 to $28^{\circ}C$ for about 3 weeks with 12-hour light per day.

Assessing Productivity of Elementary School Lunch Foodservices in Daegu and Gyeongsangbuk-do Area (대구ㆍ경북지역 초등학교 급식소의 급식생산성 분석)

  • 박영숙
    • Korean journal of food and cookery science
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    • v.19 no.3
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    • pp.286-294
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    • 2003
  • The purpose of this study was to analyze the food service management practices and productivity in 49 elementary schools in the Daegu and Gyeongsangbuk-do areas. Survey questionnaires were used to obtain a variety of quantitative and qualitative information, including general food service management and productivity, on elementary school food service systems. Descriptive analysis, $\div$2-test, t-test and one-way ANOVA analysis were used as the statistical methods in this study. Eighteen elementary schools were located in urban areas, 13 in provincial areas and 18 in isolated areas. The average number of meals was 565.1, with a significant difference (p=0.001) between the areas. The average cost per a meal was 1151.0 Won, with a significant difference (p=0.001) between the areas. The productivity Index (meal/hour) was 13.5, with a significant difference (p=0.001) between the areas. There was a significant positive correlation of the productivity Index between the total number of meal, the number of employees, the total food cost, meals per employee and the employee's working period. There was a significant negative correlation between the productivity Index and the number of side dishes, the lost per meal, the labor cost per meal and the employee's job satisfaction degree index (JDI).

Optimization of Medium Components using Response Surface Methodology for Cost-effective Mannitol Production by Leuconostoc mesenteroides SRCM201425 (반응표면분석법을 이용한 Leuconostoc mesenteroides SRCM201425의 만니톨 생산배지 최적화)

  • Ha, Gwangsu;Shin, Su-Jin;Jeong, Seong-Yeop;Yang, HoYeon;Im, Sua;Heo, JuHee;Yang, Hee-Jong;Jeong, Do-Youn
    • Journal of Life Science
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    • v.29 no.8
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    • pp.861-870
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    • 2019
  • This study was undertaken to establish optimum medium compositions for cost-effective mannitol production by Leuconostoc mesenteroides SRCM201425 isolated from kimchi. L. mesenteroides SRCM21425 from kimchi was selected for efficient mannitol production based on fructose analysis and identified by its 16S rRNA gene sequence, as well as by carbohydrate fermentation pattern analysis. To enhance mannitol production by L. mesenteroides SRCM201425, the effects of carbon, nitrogen, and mineral sources on mannitol production were first determined using Plackett-Burman design (PBD). The effects of 11 variables on mannitol production were investigated of which three variables, fructose, sucrose, and peptone, were selected. In the second step, each concentration of fructose, sucrose, and peptone was optimized using a central composite design (CCD) and response surface analysis. The predicted concentrations of fructose, sucrose, and peptone were 38.68 g/l, 30 g/l, and 39.67 g/l, respectively. The mathematical response model was reliable, with a coefficient of determination of $R^2=0.9185$. Mannitol production increased 20-fold as compared with the MRS medium, corresponding to a mannitol yield 97.46% when compared to MRS supplemented with 100 g/l of fructose in flask system. Furthermore, the production in the optimized medium was cost-effective. The findings of this study can be expected to be useful in biological production for catalytic hydrogenation causing byproduct and additional production costs.

A Study on the Application of Other Effective Area-based Conservation Measures(OECMs) for Natural Heritage - Focusing on the Old Big Trees of Natural Monument and Dangsan Ritual - (자연유산의 '기타 효과적인 지역기반 보전수단(OECMs)' 등재기준 적용 연구 - 천연기념물 노거수와 당산제를 중심으로 -)

  • Jun, Da-Seul;Shin, Hyun-Sil
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.40 no.3
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    • pp.1-9
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    • 2022
  • This study compared and reviewed the recognition determinants by applying the OECMs criteria, focusing on old big trees, plant of natural monument that are natural heritage under the national heritage system of the Cultural Heritage Administration, and the results are as follows. First, among the protected areas designated and managed by government agencies according to each protection purpose, it is necessary to actively introduce new conservation measures, OECMs, to fulfill the Biodiversity strategy for 2030 while the land area is already saturated. Second, the OECMs are geographically defined areas(CBD, 2018), not currently recognized as a protected areas, governed and managed in a way that achieves positived sustained and effective contribution to in situ conservation of biodiversity. Since the selection of term, the scope of application criteria, and the context of interpretation are inevitably different, it is necessary to separately legislate and establish related laws of the OECMs suitable for each country's situation. Third, as a result of reviewing the OECMs criteria for plant of natural monument, the final 58 potential resources were recognized. Important elements among the OECMs criteria are that buffer zones should be spaced apart from designated zones to secure a certain area, and that economic activities through commercial production should not occur and meet biodiversity standards. Among the potential candidates, 23 areas were analyzed to be geographically isolated and independent, such as Forest of Oriental Arborvitae in Do-dong, Daegu, and forest types such as Carstor Aralia of Gungchon-ri, Samcheok and Forest of Common Camellias in Maryang-ri, Seocheon. As a result of reviewing the application of OECMs criteria for plant of natural monument, it was confirmed that the functions as a traditional uses were specialized among the values of biodiversity, and ecosystem services and cultural and spiritual values were inherited through Korea's unique culture of old big trees and Dangsan ritual. In terms of biodiversity criteria, it can be used as an important factor in connecting human and natural ecosystem networks without the discovery of new species.