• Title/Summary/Keyword: Time-dependent analysis

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Application of a New Conjugation Method to Fish Pathogenic Bacteria Containing R Plasmid for the Analysis of Drug-Resistant Status in Aquaculture (새로운 conjugation 방법을 응용한 R plasmid 함유 어병세균의 분리와 양식장 내성균의 현황 분석)

  • Yoo Min Ho;Jeong Joon Beom;Kim Eun Heui;Lee Hyoung Ho;Jeong Hun Do
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.35 no.2
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    • pp.115-121
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    • 2002
  • To develop a new method of conjugation and to determine the distribution of R plasimds, we isolated multi-drug resistant strains from fish pathogenic bacteria in the farms of south and east seacoasts of Korea. Out of the 134 isolates examined, 10 showed resistance to chloramphenicol, tetracycline, streptomycin, ampicillin, colistin, nalidixic acid, oxolinic acid and kanamycin. One out of 10 multi-drug resistance bacteria, Vibfio damsela JE1 (V. damsela JE1), contained transferable R plasmid of chlorarnphenicol- tetracycline resistance genes and other nucleic acids encoding ampicillin and kanamycin resistance. The presence of the R plasmid was confirmed by conjugation using the chromocult medium (CC) as a selective and differential medium for transconiugants with identification based on the growth or colors of the colonies. The frequency of R plasmid transfer with filter mating method was come out much higher than that of broth mating method and appeared to be dependent upon the mating time and temperature. The optimum conditions for filter mating method were found to be 30$^{\circ}C$ and 24hrs as mating temperature and period, respectively, Moreover, donor cells with R plasmid, both isolate and standard bacteria, were shown to have an ability to transfer the plasmid against Escherichia coli K-12 HB101 (E. coli HB101) and Edwardsiella tarda (E. tarda) RE14 at fairly high frequencies, finally, we isolated 3 isolates of Sphingomonas sp., carrying R plasmid from 12 multi-drug resistant bacteria in normal microflora of the flounder (Paralichthys olivaceus) group used for the isolation of V emsela JE1 four months before. The same size and gene transfer chayateristics of R plasimds with those of V damsela JE1 confirmed that normal microflora have the reservoir activity for R plasmid in natural aquatic environment.

Processed Panax ginseng, sun ginseng, inhibits the differentiation and proliferation of 3T3-L1 preadipocytes and fat accumulation in Caenorhabditis elegans

  • Lee, Hyejin;Kim, Jinhee;Park, Jun Yeon;Kang, Ki Sung;Park, Joeng Hill;Hwang, Gwi Seo
    • Journal of Ginseng Research
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    • v.41 no.3
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    • pp.257-267
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    • 2017
  • Background: Heat-processed ginseng, sun ginseng (SG), has been reported to have improved therapeutic properties compared with raw forms, such as increased antidiabetic, anti-inflammatory, and antihyperglycemic effects. The aim of this study was to investigate the antiobesity effects of SG through the suppression of cell differentiation and proliferation of mouse 3T3-L1 preadipocyte cells and the lipid accumulation in Caenorhabditis elegans. Methods: To investigate the effect of SG on adipocyte differentiation, levels of stained intracellular lipid droplets were quantified by measuring the oil red O signal in the lipid extracts of cells on differentiation Day 7. To study the effect of SG on fat accumulation in C. elegans, L4 stage worms were cultured on an Escherichia coli OP50 diet supplemented with $10{\mu}g/mL$ of SG, followed by Nile red staining. To determine the effect of SG on gene expression of lipid and glucose metabolism-regulation molecules, messenger RNA (mRNA) levels of genes were analyzed by real-time reverse transcription-polymerase chain reaction analysis. In addition, the phosphorylation of Akt was examined by Western blotting. Results: SG suppressed the differentiation of 3T3-L1 cells stimulated by a mixture of 3-isobutyl-1-methylxanthine, dexamethasone, and insulin (MDI), and inhibited the proliferation of adipocytes during differentiation. Treatment of C. elegans with SG showed reductions in lipid accumulation by Nile red staining, thus directly demonstrating an antiobesity effect for SG. Furthermore, SG treatment down-regulated mRNA and protein expression levels of peroxisome proliferator-activated receptor subtype ${\gamma}$ ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein-alpha ($C/EBP{\alpha}$) and decreased the mRNA level of sterol regulatory element-binding protein 1c in MDI-treated adipocytes in a dose-dependent manner. In differentiated 3T3-L1 cells, mRNA expression levels of lipid metabolism-regulating factors, such as amplifying mouse fatty acid-binding protein 2, leptin, lipoprotein lipase, fatty acid transporter protein 1, fatty acid synthase, and 3-hydroxy-3-methylglutaryl coenzyme A reductase, were increased, whereas that of the lipolytic enzyme carnitine palmitoyltransferase-1 was decreased. Our data demonstrate that SG inversely regulated the expression of these genes in differentiated adipocytes. SG induced increases in the mRNA expression of glycolytic enzymes such as glucokinase and pyruvate kinase, and a decrease in the mRNA level of the glycogenic enzyme phosphoenol pyruvate carboxylase. In addition, mRNA levels of the glucose transporters GLUT1, GLUT4, and insulin receptor substrate-1 were elevated by MDI stimulation, whereas SG dose-dependently inhibited the expression of these genes in differentiated adipocytes. SG also inhibited the phosphorylation of Akt (Ser473) at an early phase of MDI stimulation. Intracellular nitric oxide (NO) production and endothelial nitric oxide synthase mRNA levels were markedly decreased by MDI stimulation and recovered by SG treatment of adipocytes. Conclusion: Our results suggest that SG effectively inhibits adipocyte proliferation and differentiation through the downregulation of $PPAR{\gamma}$ and $C/EBP{\alpha}$, by suppressing Akt (Ser473) phosphorylation and enhancing NO production. These results provide strong evidence to support the development of SG for antiobesity treatment.

Bioactive Materials and Biological Activity in the Extracts of Leaf, Stem Mixture and Root from Angelica gigas Nakai (참당귀 잎, 줄기혼합물과 뿌리 추출물의 생리활성물질 및 그 활성작용)

  • Heo, Jin-Sun;Cha, Jae-Young;Kim, Hyun-Woo;Ahn, Hee-Young;Eom, Kyung-Eun;Heo, Su-Jin;Cho, Young-Su
    • Journal of Life Science
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    • v.20 no.5
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    • pp.750-759
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    • 2010
  • The bioactive materials (phenolic compounds, flavonoids, minerals, decursin and decursinol angelate) and biological activities (DPPH [$\alpha,\alpha$'-diphenyl-$\beta$-picrylhydrazyl] free radical scavenging capability, reducing power, and tyrosinase activity) in the extracts of leaf, stem mixture (AGLS), and root (AGR) from Angelica gigas Nakai were examined by using water, hot water and ethanol solvent. The highest extract yield (21.89%) was found in the water extract of AGR. The highest concentrations of phenolic compounds and flavonoids in the ethanol extracts of AGLS and AGR were 14.99% and 14.79%. Major minerals of AGLS and AGR were K, Mg, Fe, Na and Ca. Decursin and decursinol angelate were the major ingredients of Angelica gigas, detected at 18.71 and 18.89 min of retention time by HPLC analysis, respectively. The highest concentrations of decursin and decursinol angelate in the Angelica gigas ethanol extract were found in root ($41.7\;{\mu}g/g$) and leaf ($34.04\;{\mu}g/g$). The highest free radical scavenging activity was found in the hot water extracts of AGLS and AGR, and its activity was stronger in all extracts of AGLS than AGR. The highest reducing power was found in the ethanol extracts of AGLS and AGR and this was dependent on the sample concentration. The hot water extracts of AGLS and AGR revealed the highest inhibition activity on tyrosinase. Overall, these results may provide the basic data needed to understand the biological activities of bioactive materials derived from Angelica gigas.

Anti-atopic Effects of Castanea crenata Inner Shell Extracts Fermented by Lactobacillus bifermentans (Lactobacillus bifermentans로 발효한 율피의 항아토피 효과)

  • Kim, Bae Jin;Son, Woo Rim;Choi, Mi Ok;Jo, Seung Kyeung;Jung, Hee Kyoung;Lee, Jin Tae;Kim, Hak Yoon;Kwoen, Dae Jun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.9
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    • pp.1378-1386
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    • 2013
  • Atopic dermatitis (AD) is a common skin disease characterized by chronic and relapsing inflammatory dermatitis with immunological disturbances. In spite of the continuous increase in the incidence of AD, it is regrettable that till date there is no effective treatment to treat the same. Therefore, the present study was designed to examine the possible anti-atopic effects of Castanea crenata inner shell extracts fermented by Lactobacillus bifermentans (FCS) in 2,4-dinitrochlorobenzene (DNCB) induced AD in NC/Nga mice. Based on the results of HPLC analysis, we found that FCS contains anti-inflammatory factors such as gallic acid (10.18 mg/g) and ellagic acid (2.14 mg/g). The groups that we have used in this study included 0.1%, 1%, 5% fermented Castanea crenata inner shell extracts (FCS 0.1, FCS 1, FCS 5), 1,3-butylene glycol treated control (AD), and normal mice. After topical FCS treatment, we observed that the clinical severity score for AD was lower in both the FCS 1 and FCS 5 groups than the AD group. We also proved beyond doubt that there was improvement of melanin, erythema and skin moisture indices in the FCS 5 group. Spleen index and gene expression levels of pro-inflammatory cytokines such as IL-$1{\beta}$ and TNF-${\alpha}$ were significantly decreased in the FCS 5 group compared to the AD group (P<0.05). Further, we also found that the level of serum immunoglobulin E (IgE) in the FCS-treated group was decreased in a concentration-dependent manner. The results of our study suggest that FCS can be effectively used as a cosmeceutical ingredient for both the prevention and improvement of AD.

Effect of Nardostachyos Rhizoma on Apoptosis, Differentiation and Proliferation in HL-60 cells

  • Ju Sung-Min;Lee Jun;Choi Ho-Seung;Yoon Sang-Hak;Kim Sung-Hoon;Jeon Byung-Hun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.1
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    • pp.163-170
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    • 2006
  • Nardostachyos Rhizoma (N. Rhizoma) belonging to the family Valerianaceae has been anti-arrhythmic effect, and sedation to the central nerve and a smooth muscle. We reported that the water extract of N. Rhizoma induced apoptotic cell death and differentiation in human promyelocytic leukemia (HL-60) cells. Cytotoxicity of N. Rhizoma was detected only in HL-60 cells (IC50 is about 200 ${\mu}g/ml$). The cytotoxic activity of N. Rhizoma in HL-60 cells was increased in a dose-dependent manner. We used several measures of apoptosis to determine whether these processes were involved in N. Rhizoma-induced apoptotic cell death. The high-dose (200 ${\mu}g/ml$) treatment of N. Rhizoma to HL-60 cells showed cell shrinkage, cell membrane blobbing, apoptotic bodies, and the fragmentation of DNA, suggesting that these cells underwent apoptosis. Treatment of HL-60 cells with N. Rhizoma time-dependently induced activation of caspase-3, caspase-8, and caspase-9 and proteolytic cleavage of poly(ADP-ribose) polymerase. Also, we investigated the effect of N. Rhizoma on cellular differentiation and proliferation in HL-60 cells. Differentiation and proliferation of HL-60 cells was determined through expression of CD11b and CD14 surface antigens using flow cytometry and nitroblue tetrazolium (NBT) assay, and through analysis of cell cycle using propidium iodide assay, respectively. N. Rhizoma induced the differentiation of HL-60 at the low-dose (100 ${\mu}g/ml$) treatment, as shown by increased expression of differentiation surface antigen CD11b, but not CDl4 and increased reducing activity of NBT. When HL-60 cells were treated with N. Rhizoma at concentration of $50{\mu}g/ml\;and\;100{\mu}g/ml$, NBT-reducing activities induced approximately 1.5-fold and 20.0-fold as compared with the control. In contrast, HL-60 cells treated with the N. Rhizoma-ATRA combination showed markedly elevated levels of 26.3-fold at $50{\mu}g/ml$ N. Rhizoma-0.1 ${\mu}M$ ATRA combination and 27.5-fold at 50 ${\mu}g/ml$ N. Rhizoma-0.2 ${\mu}M$ ATRA combination than when treated with N. Rhizoma alone or ATRA alone. It may be that N. Rhizoma plays important roles in synergy with ATRA during differentiation of HL-60 cells. DNA flow-cytometry indicated that N. Rhizoma markedly induced a G1 phase arrest of HL-60 cells. N. Rhizoma-treated HL-60 cells increased the cell population in G1 phase from 32.71% to 42.26%, whereas cell population in G2/M and S phases decreased from 23.61% to 10.33% and from 37.78% to 33.98%, respectively. We examined the change in the $p21^{WAF1/Cip1}\;and\;p27^{Kip1}$ proteins, which are the CKIs related with the G1 phase arrest. The expression of the CDK inhibitor $p27^{Kip1},\;but\;not\;p21^{WAF1/Cip1}$ were markedly increased by N. Rhizoma. Taken together, these results demonstrated that N. Rhizoma induces apoptotic cell death through activation of caspase-3, and potently inhibits the proliferation of HL-60 cells via the G1 phase cell cycle arrest in association with $p27^{Kip1}$ and granulocytic differentiation induction .

Isolation and Characterization of a Marine Bacterium, Pseudomonas sp. YJ-1 with Anti-Methicillin Resistant Staphylococcus aureus Activity (항 Methicillin Resistant Staphylococcus aureus 활성을 가지는 해양미생물 Pseudomonas sp. YJ-1의 분리와 특성)

  • Woo, Ye-Ju;Jeong, Seong-Yun
    • Korean Journal of Environmental Biology
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    • v.35 no.4
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    • pp.694-705
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    • 2017
  • The aim of this study was to isolate and identify marine bacterium with anti-methicillin-resistant Staphylococcus aureus (MRSA) activity, and to purify the anti-MRSA compound, as well as to determine its activity and synergistic effects. Among the marine bacteria isolated in this study, the YJ-1 isolate had the strongest anti-MRSA activity. The YJ-1 isolate was identified on the basis of its biochemical characteristics and an analysis of 16S rRNA gene sequences. The YJ-1 isolate showed over 99.2% homology with Pseudomonas stutzeri, and was designated as a Pseudomonas sp. YJ-1. The optimal culture conditions were $25^{\circ}C$ and initial pH 7.0. For the purification of the anti-MRSA compounds, the YJ-1 was cultured in Pa PES-II medium, and the culture filtrates were extracted by ethyl acetate, hexane, and 80% MeOH. The 80% MeOH fraction was separated by a $C_{18}$ ODS column, silica gel chromatography and a reverse phase HPLC, to yield three anti-MRSA agents, the MR1, MR2, and MR3 compounds. When the MR1 compound of $250{\mu}g\;mL^{-1}$ concentration was applied to the MRSA cells, over 95% of bacterial cells was killed within 48 hr. Compared with vancomycin and ampicillin, the MR1 compound showed significant anti-MRSA activity. In addition, the anti-MRSA activity was increased by dose and time dependent manners. Furthermore, the combination of an MR1 compound with vancomycin produced a more rapid decrease in the MRSA cells than did the MR1 compound alone. Taken together, our results suggest that the Pseudomonas sp. YJ-1 and its anti-MRSA compounds could be employed as a natural antibacterial agent in MRSA infections.

Phosphate Concentration Dependent Degradation of Biofilm in S. aureus Triggered by Physical Properties (인산염 농도에 따른 물성 변화로 발생하는 황색포도상구균 바이오필름 제거 현상)

  • Song, Sang-Hun;Hwang, Byung Woo;Son, Seong Kil;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.47 no.4
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    • pp.361-368
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    • 2021
  • The objective of this study was to establish technology for removing bacteria with human- and eco-friendly material. Staphylococcus aureus as an important component for balanced equilibrium among microbiomes, was cultured under various concentrations of phosphate. Experimental observation relating to physical properties was performed in an addition of phosphate buffer. Statistically minimum value of size and hardness using atomic force microscope was observed on the matured biofilm at 5 mM concentration of phosphate. As a result of absorbance for the biofilm tagged with dye, concentration of biofilm was reduced with phophate, too. To identify whether this reduction by phosphate at the 5 mM is caused by counter ion or not, sodium chloride was treated to the biofilm under the same condition. To elucidate components of the biofilm counting analysis of the biofilm using time-of-flight secondary ion mass spectrometry was employed. The secondary ions from the biofilm revealed that alteration of physical properties is consistent to the change of extracellular polymeric substrate (EPS) for the biofilm. Viscoelastic characterization of the biofilm using a controlled shear stress rheometer, where internal change of physical properties could be detected, exhibited a static viscosity and a reduction of elastic modulus at the 5 mM concentration of phosphate. Accordingly, bacteria at the 5 mM concentration of phosphate are attributed to removing the EPS through a reduction of elastic modulus for bacteria. We suggest that the reduction of concentration of biofilm induces dispersion which assists to easily spread its dormitory. In conclusion, it is elucidated that an addition of phosphate causes removal of EPS, and that causes a function of antibiotic.

Perceived Social Support Among the Elderly People Living Alone and Their Preference for Institutional Care: Analysis of the Mediator Effect in the Perception of the Probability of Lonely Death (독거노인의 지각된 사회적 지지와 시설 돌봄 선호: 고독사 가능성 인식의 매개 효과 분석)

  • Cho, Hye Jin;Lee, Jun Young
    • 한국노년학
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    • v.40 no.4
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    • pp.707-727
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    • 2020
  • This study aims to empirically analyze the role that perception of the probability of lonely death among the elderly people living alone plays in the relationship between perceived social support and preference for institutional care based on Andersen's expanded Behavioral Model (2002). The subjects (n=676) of this study were the elderly people living alone, extracted from the "2018 Seoul Aging Survey." With "perceived social support" as an independent variable, "preference for institutional care" as a dependent variable, and "perception of the probability of lonely death" as a mediator variable, we conducted a Binary Logistic Regression to follow the three steps of analyzing mediation effect, as suggested by Baron and Kenny (1986). The results showed that perceived social support has a negative effect on the preference for institutional care and perception of the probability of lonely death among the elderly people living alone; at the same time, perception of the probability of lonely death was found to have a positive effect on their preference for institutional care. Lastly, perception of the probability of lonely death was found to partially mediate the effect of perceived social support among the elderly people living alone in terms of their preference for institutional care. Based on these findings, the practical implications of this study can be summarized as follows. First, various programs and support should be provided to the elderly people living alone in order to enhance the level of perceived social support, a factor that has been confirmed to increase preference for institutional care among the elderly people living alone. Second, as the perception of the probability of lonely death was confirmed to be a psychosocial factor of the preference for institutional care, we need to promote education and support for older people living alone to prepare them for lonely death. These efforts are expected to form a foundations for implementing a community-based integrated care system, "Aging in Place," which is the policy direction required for older people care.

Analysis of Determinants of Carbon Emissions Considering the Electricity Trade Situation of Connected Countries and the Introduction of the Carbon Emission Trading System in Europe (유럽 내 탄소배출권거래제 도입에 따른 연결계통국가들의 전력교역 상황을 고려한 탄소배출량 결정요인분석)

  • Yoon, Kyungsoo;Hong, Won Jun
    • Environmental and Resource Economics Review
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    • v.31 no.2
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    • pp.165-204
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    • 2022
  • This study organized data from 2000 to 2014 for 20 grid-connected countries in Europe and analyzed the determinants of carbon emissions through the panel GLS method considering the problem of heteroscedasticity and autocorrelation. At the same time, the effect of introducing ETS was considered by dividing the sample period as of 2005 when the European emission trading system was introduced. Carbon emissions from individual countries were used as dependent variables, and proportion of generation by each source, power self-sufficiency ratio of neighboring countries, power production from resource-holding countries, concentration of power sources, total energy consumption per capita in the industrial sector, tax of electricity, net electricity export per capita, and size of national territory per capita. According to the estimation results, the proportion of nuclear power and renewable energy generation, concentration of power sources, and size of the national territory area per capita had a negative (-) effect on carbon emissions both before and after 2005. On the other hand, the proportion of coal power generation, the power supply and demand rate of neighboring countries, the power production of resource-holding countries, and the total energy consumption per capita in the industrial sector were found to have a positive (+) effect on carbon emissions. In addition, the proportion of gas generation had a negative (-) effect on carbon emissions, and tax of electricity were found to have a positive (+) effect. However, all of these were only significant before 2005. It was found that net electricity export per capita had a negative (-) effect on carbon emissions only after 2005. The results of this study suggest macroscopic strategies to reduce carbon emissions to green growth, suggesting mid- to long-term power mix optimization measures considering the electricity trade market and their role.

A Study on Damage factor Analysis of Slope Anchor based on 3D Numerical Model Combining UAS Image and Terrestrial LiDAR (UAS 영상 및 지상 LiDAR 조합한 3D 수치모형 기반 비탈면 앵커의 손상인자 분석에 관한 연구)

  • Lee, Chul-Hee;Lee, Jong-Hyun;Kim, Dal-Joo;Kang, Joon-Oh;Kwon, Young-Hun
    • Journal of the Korean Geotechnical Society
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    • v.38 no.7
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    • pp.5-24
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    • 2022
  • The current performance evaluation of slope anchors qualitatively determines the physical bonding between the anchor head and ground as well as cracks or breakage of the anchor head. However, such performance evaluation does not measure these primary factors quantitatively. Therefore, the time-dependent management of the anchors is almost impossible. This study is an evaluation of the 3D numerical model by SfM which combines UAS images with terrestrial LiDAR to collect numerical data on the damage factors. It also utilizes the data for the quantitative maintenance of the anchor system once it is installed on slopes. The UAS 3D model, which often shows relatively low precision in the z-coordinate for vertical objects such as slopes, is combined with terrestrial LiDAR scan data to improve the accuracy of the z-coordinate measurement. After validating the system, a field test is conducted with ten anchors installed on a slope with arbitrarily damaged heads. The damages (such as cracks, breakages, and rotational displacements) are detected and numerically evaluated through the orthogonal projection of the measurement system. The results show that the introduced system at the resolution of 8K can detect cracks less than 0.3 mm in any aperture with an error range of 0.05 mm. Also, the system can successfully detect the volume of the damaged part, showing that the maximum damage area of the anchor head was within 3% of the original design guideline. Originally, the ground adhesion to the anchor head, where the z-coordinate is highly relevant, was almost impossible to measure with the UAS 3D numerical model alone because of its blind spots. However, by applying the combined system, elevation differences between the anchor bottom and the irregular ground surface was identified so that the average value at 20 various locations was calculated for the ground adhesion. Additionally, rotation angle and displacement of the anchor head less than 1" were detected. From the observations, the validity of the 3D numerical model can obtain quantitative data on anchor damage. Such data collection can potentially create a database that could be used as a fundamental resource for quantitative anchor damage evaluation in the future.