• Journal of Life Science
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• v.23 no.2
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• pp.197-206
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• 2013

The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.26 no.1
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• pp.163-186
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• 2000

We have previously screened 150 medicinal plants on the inhibition of elastase and found a significant inhibitory effects of the extracts of Areca catechu L. on the aging and inflammation against the skin tissues. To isolate and identify the compounds having biological activity, we was further purified by each of the solvent fractions, silica gel column chromatography, preparative TLC and reversed-Phase HPLC. Peak in HPLC, which coincided with the inhibitory activity against elastase, was identified as Phenolic substance using various colorimetric methods, UV, and IR. $IC_{50}$/ values of phenolic substance purified from Areca catechu were 26.9 $\mu\textrm{g}$/$m\ell$ for porcine pancreatic elastase (PPE) and 60.8 $\mu\textrm{g}$/$m\ell$ for human neutrophil elastase (HNE). This Phenolic substance showed more potent activity than those of reference compounds, oleanolic acid (76.5 $\mu\textrm{g}$/$m\ell$ for PPE, 219.2 $\mu\textrm{g}$/$m\ell$ for HNE) and ursolic acid (31.0 $\mu\textrm{g}$/$m\ell$ for PPE, 118.6 $\mu\textrm{g}$/$m\ell$ for HNE). According to the Lineweaver-Burk Plots, the inhibition against both PPE and HNE by this phenolic substance was competitive with substrate. Phenolic substance from Areca catechu exhibited high free radical scavenging effect ($SC_{50}$/ : 6 $\mu\textrm{g}$/$m\ell$) and inhibited effectively hyaluronidase activity ($IC_{50}$/: 210 $\mu\textrm{g}$/$m\ell$). These results suggest that the Phenolic substance Purified from Areca catechu showed anti-aging effect by protecting connective tissue proteins.

• Journal of Ginseng Research
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• v.43 no.4
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• pp.600-605
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• 2019

Background: The leaves and roots of Panax ginseng are rich in ginsenosides. However, the chemical compositions of the leaves and roots of P. ginseng differ, resulting in different medicinal functions. In recent years, the aerial parts of members of the Panax genus have received great attention from natural product chemists as producers of bioactive ginsenosides. The aim of this study was the isolation and structural elucidation of novel, minor ginsenosides in the leaves of P. ginseng and evaluation of their antiinflammatory activity in vitro. Methods: Various chromatographic techniques were applied to obtain pure individual compounds, and their structures were determined by nuclear magnetic resonance and high-resolution mass spectrometry, as well as chemical methods. The antiinflammatory effect of the new compounds was evaluated on lipopolysaccharide-stimulated RAW 264.7 cells. Results and conclusions: Two novel, minor triterpenoid saponins, ginsenoside $LS_1$ (1) and 5,6-didehydroginsenoside $Rg_3$ (2), were isolated from the leaves of P. ginseng. The isolated compounds 1 and 2 were assayed for their inhibitory effect on nitric oxide production in LPS-stimulated RAW 264.7 cells, and Compound 2 showed a significant inhibitory effect with $IC_{50}$ of $37.38{\mu}M$ compared with that of NG-monomethyl-L-arginine ($IC_{50}=90.76{\mu}M$). Moreover, Compound 2 significantly decreased secretion of cytokines such as prostaglandin $E_2$ and tumor necrosis factor-${\alpha}$. In addition, Compound 2 significantly suppressed protein expression of inducible nitric oxide synthase and cyclooxygenase-2. These results suggested that Compound 2 could be used as a valuable candidate for medicinal use or functional food, and the mechanism is warranted for further exploration.

• The Journal of Engineering Geology
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• v.31 no.4
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• pp.659-669
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• 2021

The purpose of this study is to elucidate the relationship between occurrence of natural radioactive materials such as ²³⁸U and ²²²Rn and original mixing ratio of helium isotope of groundwater from various geology, and to suggest the underground aquifer environment from helium original mixing data. 9 groundwater samples were collected from five study areas, and ²³⁸U, Rn-222 and helium isotope were analyzed. A high ²³⁸U content of the range of 218~477 ㎍ /L in the groundwater occurs in the twomica granite. ⁴He air-crust mixing ratio and the Rn-222 content show a rough relation, that is, Rn-222 content increases according to the increase of ⁴He crust mixing ratio. Because of helium and radon are an inert gas, their behavior in underground environment is assumed as an analogous. The ²³⁸U content and He isotope in groundwater does not show any distinct correlation. The groundwater can be classified as three groups (air, air-crust mixing, crust-mantle mixing origin) on the diagram of ³He/⁴He vs ⁴He/²⁰Ne, which is composed of original mixing line from air-crust-mantle end members. This original mixing of helium can provide the information of underground aquifer characteristic such as the connection with surface environment or isolation condition from air environment.

• Korean Journal of Ecology and Environment
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• v.56 no.1
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• pp.94-103
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• 2023

Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria's early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.

• Journal of Life Science
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• v.32 no.3
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• pp.202-209
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• 2022

The green alga Chlamydomonas reinhardtii, a unicellular haploid eukaryote, has long been used by researchers and industries as a cell factory to produce high value-added microalgae substances using genetic modification. Microalga K01, presumed to be Chlamydomonas, was isolated from 12 freshwater samples from the Chungcheong and Jeolla regions to replace C. reinhardtii, an introduced species currently used in most basic and industrial research. The isolated K01 strain was identified as C. reinhardtii through morphological and phylogenetic studies of the 18S rDNA gene sequence (NCBI accession number KC166137). The growth and lipid content of the isolated C. reinhardtii K01 were compared with three wild and four mutant strains in TAP medium, and it was found that the K01 strain could produce 1.74×10⁷ cells/ml by the third day of culture. The growth rate of C. reinhardtii K01 was 1.5 times faster than UTEX2244, which showed the highest number of cells (1.20×10⁷ cells/ml) among the compared strains. The lipid content of the isolated C. reinhardtii K01 (20.67%) was similar to those of the wild strains, although the fatty acid oleate C18:1 was not detected in the isolated strain but was identified in the seven others. The cell density of the isolated strain increased to 0.87 g/l during a six-day culture in BG11 medium, where nitrate (NaNO₃) was introduced as a nitrogen source, while the seven acquired strains showed almost no cell proliferation.

• Annals of Clinical Microbiology
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• v.22 no.1
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• pp.9-13
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• 2019

Background: The isolation of carbapenemase-producing Enterobacteriaceae (CPE) has become increasingly common. Continuous surveillance for these organisms is essential because their infections are closely related to outbreaks of illness and are associated with high mortality rates. The aim of this study was to develop and evaluate multiplex PCR as a means of detecting several important CPE genes simultaneously. Methods: We aimed to develop a multiplex PCR that could detect seven CPE genes simultaneously. The multiplex PCR was composed of seven primer sets for the detection of KPC, IMP, VIM, NDM-1, GES, OXA-23, and OXA-48. We designed different PCR product sizes of at least 100 bp. We evaluated the performance of this new test using 69 CPE-positive clinical isolates. Also, we confirmed the specificity to rule out false-positive reactions by using 71 carbapenem-susceptible clinical strains. Results: A total of 69 CPE clinical isolates showed positive results and were correctly identified as KPC (N=14), IMP (N=13), OXA-23 (N=12), OXA-48 (N=11), VIM (N=9), GES (N=5), and NDM (N=5) by the multiplex PCR. All 71 carbapenem-susceptible clinical isolates, including Enterococcus faecalis, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, and Pseudomonas aeruginosa, showed negative results. Conclusion: This multiplex PCR can detect seven CPE genes at a time and will be useful in clinical laboratories.

• Journal of Life Science
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• v.33 no.11
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• pp.915-922
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• 2023

Lactic acid bacteria (LAB) are widespread in a variety of environments including fermented dairy products, gastroinstetinal and urogenital tracts of human and animals, plant, soil and water. Leuconostoc mesenteroides DB3 was detected by the strongest antibacterial activities among 24 Leuconostoc strains isolated from Camellia japonica flowers. Acid tolerance of L. mesenteroides DB3 existed up to pH 2.5, but the resistance did not show at pH 2.0, which relatively excellent acid resistance existed. Bile acid tolerance was very stable within the test range to 1.2%. L. mesenteroides DB3 exhibited the optimal growth at 30℃, and showed a slight slow growth when compared with L. mesenteroides KCTC3505, which reached a stationary phase at 18 hr. The pH was changed along with the growth curve, but was maintained above pH 3.98. L. mesenteroides DB3 had higher initial antibacterial activities when compared to L. mesenteroides KCTC3505, but it showed similar activities with the standard strain after the latter part of the logarithmic growth phase. Although lactic acid production in L. mesenteroides DB3 was induced by lower amount in the initial part to the standard strain, it was exhibited by similar amounts after the late logarithmic growth phase. Muicin adhesion of L. mesenteroides DB-3 maintained superior to L. mesenteroides KCTC3505. Both strains showed excellent emulsification ability for kerosene. In summary, we evaluate that L. mesenteroides DB-3 has a high potential for application as probiotics owing to its excellent antibacterial activity, acid resistance, bile acid resistance, and muicin adhesion.

• Journal of the Korean Institute of Landscape Architecture
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• v.52 no.1
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• pp.17-33
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• 2024

This study suggests management planning of Bonguisan Neighborhood Park located on the central of Chuncheon city and highly used for citizen's rest and leisure space utilizing its vegetation structure feature. Bonguisan has been the central of the chuncheon since the period of the Three states in Korean history and consistently damaged, especially in present era, an isolation and sererance of its ecosystem has deepen for indiscreet urban development. The percentage of actual vegetation of Boinguisan Neighborhood Park is as follows: Quercus mongolica is 28.5%, Quercus mongolica - Quercus serrata is 2.1%, Pinus densiflora is 15.6%, Pinus densiflora - Quercus mongolica is 15.9%, Betula schmidtii is 1.6%, Robinia pseudoacacia is 5.9%, Pinus koraiensis is 1.6%. Quercus mongolica is distributed on the southwest, northwest, southeast side of region, Pinus densiflora is distributed on the ridge of east and southeast side of region, Betula schmidtii is distributed on the valley of northeast side region and steep slope region which is on the north side of chungwonsa temple. Pinus densiflora community (Comm. I) and Quercus acutissima - Robinia pseudoacacia community (Comm. V) is expected to undergo succession since it's categorized as Quercus spp. and Quercus mongolica community (Comm. II) and Quercus serrata-Quercus mongolica community (Comm. III), Betula schmidtii community (Comm. IV), Pinus koraiensis community (Comm. VI) is expected to maintain. Also for target vegetation and management planning, Vegetation of Bonguisan Neighborhood Park is classified as 1st Natural landscape conservation and improvement type, 2nd Ecological succession type, 3rd Unusual community conservation type, and 4th Recreation and experience type. And we suggested ecological management measure about each management types. For efficient management of Bonguisan Neighborhood Park, it is need to unify management system of it and after designating Pinus densiflora community and Betula schmidtii community which has high ecological preservation value as an ecological landscape protected area and manage it.

• Tuberculosis and Respiratory Diseases
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• v.47 no.3
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• pp.321-330
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• 1999

Background : Early diagnosis and proper antibiotic treatment are very important in the management of ventilator-associated pneumonia (VAP) because of its high mortality. Bronchoscopy with a protected specimen brush (PSB) has been considered the standard method to isolate the causative organisms of VAP. However, this method burdens consumer economically to purchase a PSB. Another useful method for the diagnosis of VAP is quantitative cultures of aspirated specimens through bronchoscopic bronchoalveolar lavage (BAL), for which the infusion of more than 120 m1 of saline has been recommended for adequate sampling of a pulmonary segment. However, occasionally it leads to deterioration of the patient's condition. We studied the diagnostic efficacy of minibronchoalveolar lavage (miniBAL), which retrieves only 25 ml of BAL fluid, in the isolation of causative organisms of VAP. Methods: We included 38 consecutive patients (41 cases) suspected of having VAP on the basis of clinical evidence, who had received antibiotics before the bronchoscopy. The two diagnostic techniques of PSB and miniBAL, which were performed one after another at the same pulmonary segment, 'were compared prospectively. The cut-off values for quantitative cultures to define causative bacteria of VAP were more than $10^3$ colony-forming units (cfu)/ml for PSB and more than $10^4$ cfu/ml for BAL. Results: The amount of instilled normal saline required to retrieve 25 ml of BAL fluid was $93{\pm}32 ml$ (mean${\pm}$SD). The detection rate of causative agents was 46.3% (19/41) with PSB and 43.9% (18/41) with miniBAL. The concordance rate of PSB and miniBAL in the bacterial culture was 85.4% (35/41). Although arterial blood oxygen saturation dropped significantly (p<0.05) during ($92{\pm}10%$) and 10 min after ($95{\pm}3%$) miniBAL compared with the baseline ($97{\pm}3%$), all except 3 cases were within normal ranges. The significantly elevated heart rate during ($l25{\pm}24$/min, p<0.05) miniBAL compared with the baseline ($1l1{\pm}22$/min) recovered again in 10 min after ($111{\pm}26$/min) miniBAL. Transient hypotension was developed during the procedure in two cases. The procedure was stopped in one case due to atrial flutter. Conclusion: MiniBAL is a safe and effective technique to detect the causative organisms of VAP.