• Korean Journal of Clinical Laboratory Science
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• v.52 no.1
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• pp.53-61
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• 2020

Cell migration is a central process for recovering from wounds triggered by physical distress besides embryogenesis and cancer metastasis. Wound healing assay is widely used as a fundamental research technique for investigation of two-dimensional cell migration in vitro. The most common approach for imitating physical wound in vitro is mechanical scratching on the surface of the confluent monolayer by using sharp materials. The iron metal pin with a suspension spring for fine adjustment of the orthogonal contact surface between the scratching point and the individual bottom of multi-well plate with planar curvatures were adopted for the creative invention of a 96-well plate wound maker. While classic tips drew diverse and zigzag scratching patterns on the confluent monolayer, our wound maker displayed synchronized linear wounds in the middle of each well of a 96-well plate that was seeded with several cell lines. Given that several types of multi-well plates commercially available are compatible with our lab-made wound maker for creating uniform scratches on the confluent monolayer for the collective cell migration in wound healing assay, it is certain that the application of this wound maker to the real-time wound healing assay in high content screening (HCS) is superior than utilization of typical polypropylene pipette tips.

• Journal of Ginseng Research
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• v.41 no.2
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• pp.169-179
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• 2017

Background: Ginsenosides are the main pharmacological components of Panax ginseng root, which are thought to be primarily responsible for the suppressing effect on oxidative stress. Methods: 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and oxygen radical absorption capacity were applied to evaluate the antioxidant activities of the ginsenosides. Human embryonic kidney 293 (HEK-293) cells were incubated with ginsenosides extracted by pulsed electric field (PEF) and solvent cold soak extraction (SCSE) for 24 h and then the injury was induced by $40{\mu}M$ $H_2O_2$. The cell viability and surface morphology of HEK-293 cells were studied using MTS assay and scanning electron microscopy, respectively. Dichloro-dihydro-fluorescein diacetate fluorescent probe assay was used to measure the level of intracellular reactive oxygen species. The intracellular antioxidant activities of ginsenosides were evaluated by cellular antioxidant activity assay in HepG2 cells. Results: The PEF extracts displayed the higher 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity and stronger oxygen radical absorption capacity (with an oxygen radical absorption capacity value of $14.48{\pm}4.04{\mu}M\;TE\;per\;{\mu}g/mL$). The HEK-293 cell model also suggested that the protective effect of PEF extracts was dose-dependently greater than SCSE extracts. Dichloro-dihydro-fluorescein diacetate assay further proved that PEF extracts are more active (8% higher than SCSE extracts) in reducing intracellular reactive oxygen species accumulation. In addition, scanning electron microscopy images showed that the HEK-293 cells, which were treated with PEF extracts, maintained more intact surface morphology. Cellular antioxidant activity values indicated that ginsenosides extracted by PEF had stronger cellular antioxidant activity than SCSE ginsenosides extracts. Conclusion: The present study demonstrated the antioxidative effect of ginsenosides extracted by PEF in vitro. Furthermore, rather than SCSE, PEF may be more useful as an alternative extraction technique for the extraction of ginsenosides with enhanced antioxidant activity.

• Journal of the Korean Wood Science and Technology
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• v.35 no.1
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• pp.17-26
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• 2007

This study was conducted to investigate the effects of End-coating (CO) treatment and Vapor-dam (V) treatment on the air circulating oven drying characteristics of green stock (Paulownia tomentosa) for drum. The reduction of the weight of specimens was greatly displayed in order of control (C-NC) treatment, End-coating (C-CO) treatment, Vapor-dam (V-NC) treatment and Vapor-dam/End-coating (V-CO) treatment after the beginning of drying. The V-CO treated specimen showed smoother temperature gradient compared with the V-NC treated specimen throughout whole drying stage and there was little temperature gradient between the inner and outer part of the cylindrical stock. The C-CO treated specimen showed higher value of vapor pressure in both the inner and outer part of the cylindrical stock until the middle stage of drying, while after the middle stage of drying only the inner part of the cylindrical stock presented higher value compared with the C-NC treated specimen. The distribution of the absolute vapor pressure of the V-CO treated specimen was similar to that of the V-NC treated specimen during the initial stage of drying. However, the former showed a large distribution in order of the hollow, the outer part, the inner part of the cylindrical stock after the initial stage of drying, which was entirely different model of distribution from that of the V-NC treated specimen. Surface checks and ring failures were not observed for all of the specimens, while end checks were severely occurred for the C-NC treated specimen and the V-NC treated specimen.

• Clinical and Experimental Reproductive Medicine
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• v.32 no.2
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• pp.133-147
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• 2005

Objectives: This study was carried out to establish human embryonic stem cells derived from frozen-thawed embryos using mouse embryonic fibroblasts (mEFs), human fetal skin and muscle fibroblasts as feeder cells, and to identify the characteristic of embryonic stem cells. Methods: When primary mEFs, human fetal skin and muscle fibroblasts were prepared, passaging on 4 days from replating could have effective trypsinization and clear feeder layers. Eight of 23 frozenthawed 4~8 cell stage embryos donated from consenting couples developed to blastocysts. Inner cell mass (ICM) was isolated by immunosurgery. ICM was co-cultured on mEFs, human fetal skin or muscle fibroblasts. The ICM colonies grown on mEFs, human fetal skin or muscle fibroblasts were tested the expression of stage specific embryonic antigen-3, -4 (SSEA-3, -4), octamer binding transcription factor-4 mRNA (Oct-4) and alkaline phosphatase surface marker. Results: We obtained 1 ICM colony from 2 ICM co-cultured on mEFs as feeder cells and did not obtain any ICM colony from 6 ICM clumps co-cultured on human fetal skin or muscle fibroblasts. The colony formed on mEFs could be passaged 30 times every 5 days with sustaining undifferentiated colony appearance. When the colonies cultured on mEFs were grown on human fetal skin or muscle fibroblasts, the colonies could be passaged 15 times every 9 days with sustaining undifferentiated colony appearance. The colonies grown on mEFs and human fetal fibroblasts expressed SSEA-4 and alkaline phosphatase surface markers and positive for the expression of Oct-4 by reverse transcription-polymerase chain reaction (RT-PCR). The produced embryoid body differentiated spontaneously to neural progenitorlike cells, neuron-like cells and beating cardiomyocyte-like cells, and frozen-thawed embryonic stem cells displayed normal 46,XX karyotype. Conclusions: The human embryonic stem cells can be established by using mEFs and human fetal fibroblasts produced in laboratory as feeder cells.

• Journal of Microbiology and Biotechnology
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• v.29 no.1
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• pp.160-170
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• 2019

The lactic acid bacteria species Lactobacillus plantarum (L. plantarum) has been used extensively for vaccine delivery. Considering to the critical role of dendritic cells in stimulating host immune response, in this study, we constructed a novel CD11c-targeting L. plantarum strain with surface-displayed variable fragments of anti-CD11c, single-chain antibody (scFv-CD11c). The newly designed L. plantarum strain, named 409-aCD11c, could adhere and invade more efficiently to bone marrow-derived DCs (BMDCs) in vitro due to the specific interaction between scFv-CD11c and CD11c located on the surface of BMDCs. After incubation with BMDCs, the 409-aCD11c strain harboring a eukaryotic vector pValac-GFP could lead to more efficient expression of GFP compared with wild-type strains shown by flow cytometry analysis, indicating the enhanced translocation of pValac-GFP from L. plantarum to BMDCs. Similar results were also observed in an in vivo study, which showed that oral administration resulted in efficient expression of GFP in both Peyer's patches (PP) and mesenteric lymph nodes (MLNs) within 7 days after the last administration. In addition, the CD11c-targeting strain significantly promoted the differentiation and maturation of DCs, the differentiation of $IL-4^+$ and $IL-17A^+$ T helper (Th) cells in MLNs, as well as production of $B220^+$ $IgA^+$ B cells in the PP. In conclusion, this study developed a novel DC-targeting L. plantarum strain which could increase the ability to deliver eukaryotic expression plasmid to host cells, indicating a promising approach for vaccine study.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.4
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• pp.557-566
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• 2005

In preparation of steamed foam-cakes, effects of whipping time, amount of wheat flour, and amounts of emulsifier on physical properties of the steamed foam cakes were investigated using RSM (response surface methodology). The three independent variables selected for the RSM experiment were whipping time $(X_1)$, amount of wheat flour $(X_2)$, and amounts of wheat flour $(X_2)$, and concentration of emulsifier $(X_3)$ were set for single-stage mixing, respectively. A rotatable central composite design was used for treatment arrangement. The responses from the product for loaf volume, color values and textural properties were analysed. In the analysis of variance for the foam cakes prepared by single-stage method, significant interactions were observed between independent variables (experimental factors) and physical property like loaf volume (p<0.05); textural properties like hardness, gumminess, and chewiness (p<0.05). Among independent variables, concentration of emulsifier had the most effects on physical properties while whipping time. The ordinary points in surface response showed maximal points with physical property like colorimetric b value while other properties revealed saddle points. The 3-dimensional response surface graphs of the predicted regression models displayed decreasing loaf volumes with increasing whipping times and emulsifier concentrations beyond optimum levels. The optimum conditions for best loaf volume and textural property (hardness, gummimess and chewiness) of the products selected by extracting intersectional areas of the contour maps that commonly overlapped all characteristics were; $11\~13$ min whipping time, $470\~486\;g$ amount of wheat flour, and $19\~20\;g$ emulsifier concentration, in case of single-stage method. The median values extracted from the RSM experimental results for optimum manufacturing conditions for single-stage method, i.e., 12 min whipping time, 478 g amount of wheat flour, and 20 g emulsifier concentration were empirically proven to fit the predicted levels of physical properties from the final foam cakes.

• Journal of the Korean Geotechnical Society
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• v.29 no.9
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• pp.5-15
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• 2013

In this paper, the unsaturated slope stability analysis considering suction stress (Lu and Godt, 2008) was introduced and the results applied for a certain sand slope were analyzed. The unsaturated slope stability analysis considering suction stress can analyze both conditions of steady infiltration and no infiltration, and it can estimate the safety factor of slope as a function of soil depth. Also, the influence of weathering phenomenon at a certain depth from the ground surface can be considered. The stability analysis considering suction stress was applied to the unsaturated infinite slope composed of sand with the relative density of 60%. The suction stress under no infiltration condition was affected by ground water table until a certain influencing depth. However, the suction stress under steady infiltration condition was affected by seepage throughout the soils. Especially, the maximum suction stress was displayed around ground surface. The factor of safety in the infinite slope under no infiltration condition rapidly increased and decreased within the influence zone of ground water table. As a result of slope stability analysis, the factor of safety is less than 1 at the depth of 2.4 m below the ground surface. It means that the probability of slope failure is too high within the range of depths. The factor of safety under steady infiltration condition is greater than that under no infiltration condition due to the change of suction stress induced by seepage. As the steady infiltration rate of precipitation was getting closer to the saturated hydraulic conductivity, the factor of safety decreased. In case of the steady infiltration rate of precipitation with $-1.8{\times}10^{-3}cm/s$, the factor of safety is less than 1 at the depths between 0.2 m and 3 m below the ground surface. It means that the probability of slope failure is too high within the range of depths, and type of slope failure is likely to be shallow landslides.

• Korean Journal of Heritage: History & Science
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• v.43 no.3
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• pp.306-333
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• 2010

Previous studies on Wolseong fortress have focused on capital system of Silla Dynasty and on the recreation of Wolseong fortress due to the excavations in and around Wolseong moat. Since the report on the Geographical Survey of Wolseong fortress was published and GPR survey in Wolseong fortress was executed as a trial test in 2004, the academic interest in the site has now expanded to the inside of the fortress. From such context, the preliminary research on the fortress including geophysical survey had been commenced. GPR survey had been conducted for a year from March, 2007. The principal purpose of the recent 3D GPR survey was to provide visualization of subsurface images of the entire Wolseong fortress area. In order to obtain 3D GPR data, dense profile lines were laid in grid-form. The total area surveyed was $112,535m^2$. Depth slice was applied to analyse each level to examine how the layers of the remains had changed and overlapped over time. In addition, slice overlay analysis methodology was used to gather reflects of each depth on a single map. Isolated surface visualization, which is one of 3D analysis methods, was also employed to gain more in-depth understanding and more accurate interpretations of the remain The GPR survey has confirmed that there are building sites whose archaeological features can be classified into 14 different groups. Three interesting areas with huge public building arrangement have been found in Zone 2 in the far west, Zone 9 in the middle, and Zone 14 in the far east. It is recognized that such areas must had been used for important public functions. This research has displayed that 3D GPR survey can be effective for a vast area of archaeological remains and that slice overlay images can provide clearer image with high contrast for objects and remains buried the site.

• The Journal of Korean Academy of Prosthodontics
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• v.49 no.3
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• pp.245-253
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• 2011

Purpose: The aim of this in vitro study was to estimate surface characteristic after peptide coating and investigate biological response of human mesenchymal stem cell to anodized titanium discs coated with RGD peptide by physical adhesion and chemical fixation. Materials and methods: Fluorescence isothiocyanate (FITC) modified RGD-peptide was coated on the anodized titanium discs (diameter 12 mm, height 3 mm) using two methods. One was physical adhesion method and the other was chemical fixation method. Physical adhesion was performed by dip and dry procedure, chemical fixation was performed by covalent bond via silanization. In this study, human mesenchymal stem cell was used for experiments. The experiments consisted of surface characteristic evaluation after peptide coating, analysis about cell adhesion, proliferation, differentiation, and mineralization. Obtained data are statistically treated using Kruskal-Wallis test and Bonferroni test was performed as post hoc test (P=.05). Results: The evaluation of FE-SEM images revealed no diffenrence at micro-surfaces between each groups. Total coating dose was higher at physical adhesion experimental group than at chemical fixation experimental group. In cell adhesion and proliferation, RGD peptide coating did not show a statistical significance compared with control group (P>.05). In cell differentiation and mineralization, physical adhesion method displayed significantly increased levels compared with control group and chemical fixation method (P<.05). Conclusion: RGD peptide coating seems to enhance osseointegration by effects on the response of human mesenchymal stem cell. Especially physical adhesion method showed more effective than chemical fixation method on response of human mesenchymal stem cell.

• Korean Journal of Environmental Biology
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• v.24 no.1 s.61
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• pp.67-80
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• 2006

The effect of environmental forcing on picophytoplankton distribution pattern was investigated in the tropical and subtropical western Pacific (TSWP) and the East Sea in September, 2002, and the continental shelf of the East China Sea (C-ECS) in August, 2003. The abundance of picophytoplankton populations, Synechococcus, Prochlorococcus and picoeukaryotes were determined by flow cytometry analyses. Picophytoplankton vertical profiles and integrated abundance $(0\sim100\;m)$ were compared with these three physiochemically different regions. Variation patterns of integrated cell abundance of Synechococcus and Prochlorococcus in these three regions showed contrasting results. Synechococcus showed average abundance of $84.5X10^{10}\;cells\;m^{-2}$, in the TSWP, $305.6X10^{10}\;cells\;m^{-2}$ in the C-ECS, and $125.4X10^{10}\;cells\; m^{-2}$ in the East Sea where increasing cell concentrations were observed in the region with abundant nutrient. On the other hand, Prochlorococcus showed average abundance of $504.5X10^{10}\;cells\;m^{-2}$ in the TSWP, $33.2x10^{10}\;cells\;m^{-2}$ in the C-ECS, and $130.2X10^{10}\;cells\;m^{-2}$ in the East Sea exhibiting a distinctive pattern of increasing cell abundance in oligotrophic warm water. Although picoeukaryotes showed a similar pattern to Synechococcus, the abundance was 1/10 of Synechococcus. Synechococcus and picoeukaryotes showed ubiquitous distribution whereas Prochlorococcus generally did not appear in the C-ECS and the East Sea with low salinity environment. The average depth profiles for Synechococcus and Prochlorococcus displayed uniform abundance in the surface mixed layer with a rapid decrease below the surface mixed layer. for Prochlorococcus, a similar rapid decreasing trend was not observed below the surface mixed layer of the TSWP, but Prochlorococcus continued to show high cell abundance even down to 100 m depth. Picoeukaryotes showed uniform abundance along $0\sim100\;m$ depth in the C-ECS, and abundance maximum layer appeared in the East Sea at $20\sim30\;m$ depth.