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Purification and Characterization of Degradative Enzyme of Dental Plaque from Streptomyces sp. Y9343 (Streptomyces sp. Y9343이 生産하는 齒面細菌膜 分解酵素의 精製와 特性)

  • Kim, Seong-Joo;Han, Hong-Keun;Yoon, Jeong-Weon
    • Microbiology and Biotechnology Letters
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    • v.24 no.1
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    • pp.9-18
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    • 1996
  • Streptococcus mutans has been implicated as primary causative agents of dental caries by insoluble glucan (IG) in human and experimental animals. An attempt was made to search for the ${\alpha}$-1,3 glucanase that degrades IG produced by S. mutans. ${\alpha}$-1,3 glucanase was detected in the culture supernatant of microorganisms, which are isolated from soils on agar medium containing IG as a sole carbon source. This Streptomyces sp. hydrolysed IG produced by immobilized S. mutans and was named as Y9373. This enzyme required ${\alpha}$-1,3 glucan (IG) as an inducer. The optimum conditions for enzyme production were studied. The enzyme was purified by 30~70% $(NH_4)_2SO_4$ precipitation, anion exchange chroma tography on DEAE-cellulose and gel filtration on Sepadex G-75. The purified enzyme has a specific activity of 7840.0 U/mg protein giving 32.1-fold purification and final yield of 0.53%. The molecular weight was estimated to be about 22.5 kDa by SDS-PAGE. The optimum pH and temperature for enzyme reaction were 6.5 and 37$^{\circ}C$, respectively and the enzyme was relatively stable at the temperature below 60$^{\circ}C$. The activity of purified enzyme was enhanced by adding $Co^{2+},\;Mn^{2+}\;and\;Mg^{2+}$ into the medium, whereas inhibited by adding $Hg^{2+},\;Zn^{2+}$ and SDS. The $K_m\;and\;V_{max}$ value of ${\alpha}$-1,3 glucanase for IG were estimated to be 2.50 mM and 0.0431 mM/min, respectively. The thin layer chromatographic analysis of hydrolysates from IG with ${\alpha}$-1,3 glucanase showed that glucose was the main product of reaction. This enzyme activity was about 14 times higher than marketing dextranase as preventive agent against artificial dental caries by S. mutans in TH medium including 5% sucrose after 30 minutes.

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Expression and Function of Calcineurin in Inflammatory Arthritis (류마티스 관절염에서 칼시뉴린의 발현과 기능)

  • Park, Bo-Hyoung;Yoo, Seung-Ah;Hong, Kyung-Hee;Hyoung, Bok-Jin;Hwang, Yu-Na;Cho, Chul-Soo;Park, Won;Kim, Wan-Uk
    • IMMUNE NETWORK
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    • v.6 no.1
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    • pp.33-42
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    • 2006
  • Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.

Assessing the Impact of Climate Change on Water Resources: Waimea Plains, New Zealand Case Example

  • Zemansky, Gil;Hong, Yoon-Seeok Timothy;Rose, Jennifer;Song, Sung-Ho;Thomas, Joseph
    • Proceedings of the Korea Water Resources Association Conference
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    • 2011.05a
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    • pp.18-18
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    • 2011
  • Climate change is impacting and will increasingly impact both the quantity and quality of the world's water resources in a variety of ways. In some areas warming climate results in increased rainfall, surface runoff, and groundwater recharge while in others there may be declines in all of these. Water quality is described by a number of variables. Some are directly impacted by climate change. Temperature is an obvious example. Notably, increased atmospheric concentrations of $CO_2$ triggering climate change increase the $CO_2$ dissolving into water. This has manifold consequences including decreased pH and increased alkalinity, with resultant increases in dissolved concentrations of the minerals in geologic materials contacted by such water. Climate change is also expected to increase the number and intensity of extreme climate events, with related hydrologic changes. A simple framework has been developed in New Zealand for assessing and predicting climate change impacts on water resources. Assessment is largely based on trend analysis of historic data using the non-parametric Mann-Kendall method. Trend analysis requires long-term, regular monitoring data for both climate and hydrologic variables. Data quality is of primary importance and data gaps must be avoided. Quantitative prediction of climate change impacts on the quantity of water resources can be accomplished by computer modelling. This requires the serial coupling of various models. For example, regional downscaling of results from a world-wide general circulation model (GCM) can be used to forecast temperatures and precipitation for various emissions scenarios in specific catchments. Mechanistic or artificial intelligence modelling can then be used with these inputs to simulate climate change impacts over time, such as changes in streamflow, groundwater-surface water interactions, and changes in groundwater levels. The Waimea Plains catchment in New Zealand was selected for a test application of these assessment and prediction methods. This catchment is predicted to undergo relatively minor impacts due to climate change. All available climate and hydrologic databases were obtained and analyzed. These included climate (temperature, precipitation, solar radiation and sunshine hours, evapotranspiration, humidity, and cloud cover) and hydrologic (streamflow and quality and groundwater levels and quality) records. Results varied but there were indications of atmospheric temperature increasing, rainfall decreasing, streamflow decreasing, and groundwater level decreasing trends. Artificial intelligence modelling was applied to predict water usage, rainfall recharge of groundwater, and upstream flow for two regionally downscaled climate change scenarios (A1B and A2). The AI methods used were multi-layer perceptron (MLP) with extended Kalman filtering (EKF), genetic programming (GP), and a dynamic neuro-fuzzy local modelling system (DNFLMS), respectively. These were then used as inputs to a mechanistic groundwater flow-surface water interaction model (MODFLOW). A DNFLMS was also used to simulate downstream flow and groundwater levels for comparison with MODFLOW outputs. MODFLOW and DNFLMS outputs were consistent. They indicated declines in streamflow on the order of 21 to 23% for MODFLOW and DNFLMS (A1B scenario), respectively, and 27% in both cases for the A2 scenario under severe drought conditions by 2058-2059, with little if any change in groundwater levels.

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Establishment and Maintenance of Embryonic Stem-like Cell Lines from In Vitro Produced Bovine Blastocysts (체외수정 유래 소 배반포로부터 유사 배아 줄기 세포의 확립 및 유지)

  • Lee, Yu-Yeon;Kim, Sun-Uk;Kim, Ji-Su;Song, Bong-Seok;Cho, Yoon-Jeong;Park, Jung-Sun;Yu, Dae-Yeul;Jin, Dong-Il;Lee, Kyung-Kwang;Koo, Deog-Bon
    • Reproductive and Developmental Biology
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    • v.31 no.3
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    • pp.215-220
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    • 2007
  • This study was conducted to examine the establishment of bovine ES-like cells having pluripotency. The hatched blastocysts derived from culture of in vitro fertilized embryos for 10 to 12 days dissociated mechanically into ICM-and trophectoderm-rich clumps using needle, and cultured onto mitotically-inactivated MEF feeder layer. The primary colonies originated from ICM cells were detached mechanically 7 days after seeding and subsequent subculture was conducted at intervals of every 5 to 7 days. Two ES -like cell lines were established and maintained over 40 passages. Self-renewal of the established lines was confirmed by examining the alkaline phosphatase activity, stem cell-specific marker profiles including SSEA isotopes, Oct-4 and STAT3. Moreover, the established cell lines could produce anchorage-independent embryoid bodies (EBs) with gradual decrease of Oct-4 transcript level in time-dependent manner.

Fresh Water Injection Test in a Fractured Bedrock Aquifer for the Mitigation of Seawater Intrusion (해수침투 저감을 위한 균열암반 대수층 내 담수주입시험)

  • Shin, Je-Hyun;Byun, Joong-Moo
    • Economic and Environmental Geology
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    • v.43 no.4
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    • pp.371-379
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    • 2010
  • Fresh water injection test in a fractured bedrock aquifer was applied as an efficient approach to lower saline concentrations in the saltwater-freshwater transition zone formed by seawater intrusion in a coastal area. The methodology and effectiveness of fresh water injection for hydraulically controlling seawater intrusion is overwhelmingly site dependent, and there is an urgent need to characterize the permeable fractures or unconsolidated porous formations which can allow for seawater flow and transport. Considering aquifer characteristics, injection and monitoring boreholes were optimally designed and completed to inject fresh water through sand layer and fractured bedrock, respectively. We devised and used the injection system using double packer for easy field operation and maintenance. Overall fracture distribution was systematically identified from borehole image logs, and the section of fresh water injection was decided from injection test and monitoring. With fresh water injection, the fluid electrical conductivity of the monitoring well started to be lowered by the inflow of fresh water at the specific depth. And this inflow leaded to the replacement of the fluid in the upper parts of the borehole with fresh water. Furthermore, the injection effect lasted more than several months, which means that fresh water injection may contribute to the mitigation of seawater intrusion in a coastal area.

The Infection of Heterosporis anguillarum in Cultured Shortfin Eel (Anguilla bicolor pacifica) (양식동남아산 뱀장어(Anguilla bicolor pacifica)의 Heterosporis anguillarum 감염)

  • Kim, Jin-Do;Do, Jeong-Wan;Choi, Hye-Sung;Jo, Hyae-In;Lee, Nam-Sil;Kim, Young-Dae
    • Korean Journal of Environmental Biology
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    • v.32 no.4
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    • pp.382-388
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    • 2014
  • Shortfin eel (Anguilla bicolor pacifica) is a species of commercial importance and its production is greatly affected due to the infection by Heterosporis anguillarum. In this study, we evaluated the effect of H. anguillarum infection on the growth of Shortfin eel. A disease that trunk muscle of cultured shortfin eel, Anguilla bicolor pacifica, were irregular and resulted in death, breakout of the commercial eel culture farm. We observed that the trunk muscle of infected eels were irregular and represented white or yellowish externally. Histopathologically, a great numbers of large or small spores and sporophorocysts were also observed in degenerated muscle layer. The cloning of specific gene of H. anguillarum, encoding small subunit ribosomal RNA (SSU-rRNA) was amplified by the polymerase chain reaction(PCR) from the muscle lesion of diseased eel. The size of clone gene is well matched with the size of small subunit ribosomal RNA of H. anguillarum and thus confirming the infection by H. anguillarum.

A Study on the Micellization of Sodium Dodecyl Sulfate in 𝜔-Phenylakylammonium Salt Solution (𝜔-Phenylalkylammonium Salt의 수용액에서 Sodium Dodecyl Sulfate의 미셀에 관한 연구)

  • Ryu, Yeanho;Oh, Jung Hee
    • Applied Chemistry for Engineering
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    • v.8 no.6
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    • pp.1041-1047
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    • 1997
  • The influence of $\omega$-phenylalkylammonium salt on the critical micelle concentration (CMC) of SDS has been examined using the electric conductivity method. CMC of SDS exhibited the tendency to decrease with the length of alkyl group of additives. The effect of temperature on CMC of SDS in additive solutions has been observed in the range of $18^{\circ}C-50^{\circ}C$. The free energy(${\Delta}G_m^{\circ}$) for the micellization of SDS is negative and the entropy(${\Delta}S_m^{\circ}$) is a large positive value. The enthalpy(ΔHm0is positive in low temperature($18^{\circ}C$) and negative in high temperature($>25^{\circ}C$). In the prensence of organic additives, the micellization of SDS was considered as a spontaneous process and to involve a phase transition. The values of ΔGm0has shown the tendency to increase but the values of ${\Delta}S_m^{\circ}$ and ${\Delta}H_m^{\circ}$ to decrease with the length of alklyl group of additive salts. The changes in ${\Delta}\kappa$(difference of specific conductivity) with increasing mole ratio of additives in the mixed solutions indicated the formation of mixed micelles between SDS and additives. The effect of the length of alkyl chain on the micellization of SDS demonstrated the penetration of organic additives into the palisade layer of the SDS micelle.

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Estimation of Applicability of Empirical Design Procedure for Predicting Seismic Response of Buried Gas Pipelines through 3D Time-history Analysis (3차원 시간이력해석을 통한 매설가스배관 종방향 지진응답 예측을 위한 경험적 설계법의 적용성 평가)

  • Kwak, Hyungjoo;Park, Duhee;Lee, Jangguen;Kang, Jaemo
    • Journal of the Korean Geotechnical Society
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    • v.31 no.9
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    • pp.53-68
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    • 2015
  • Longitudinal strain is an important component of seismic design for buried pipelines. A design procedure which determines the wavelength from site natural period and shear wave velocity of the soil layer and closed-form solutions of pipelines under a harmonic motion is typically used in design. However, the applicability of the procedure has not yet been thoroughly investigated. In this paper, displacement-time histories extracted from 1D site response analyses are used in 3D shell-spring model to accurately predict the response of pipelines. The results are closely compared to those from the design procedure. The area of interest is East Siberia. Performing a site response analysis to determine site specific displacement time history is highlighted. The site natural period may be used to predict the predominant period of the acceleration time history, but cannot be used to estimate the predominant period of the displacement time history. If an accurate estimate of the predominant period of the displacement time history is provided, it is demonstrated that the design equation can be successfully used to predict the response of pipelines.

Effect of Vapor-Cooled Heat Stations in a Cryogenic Vessel (극저온액체 저장용기에서 열전도 차폐단의 영향)

  • Kim, S.Y.;Kang, B.H.;Choi, H.J.
    • Transactions of the Korean hydrogen and new energy society
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    • v.9 no.4
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    • pp.169-176
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    • 1998
  • An experimental study on effect of vapor-cooled heat stations in a 5.5 liter cryogenic vessel has been performed. The cryogenic vessel is made of stainless steel of thickness of 1mm and insulated by the combined insulation of vacuum, MLI(multi-layer insulation) and vapor-cooled radiation shield. Vapor-cooled heat stations are also constructed based on the 1-dimensional thermal analysis to reduce the heat inleak through a filling tube. Thermal analysis indicates that the vapor-cooled heat stations can substantially enhance the performance of vessel for cryogenic fluids with high $C_p/h_{fg}$ where $C_p$ the specific heat and $h_{fg}$ the heat of vaporization, such as $LH_2$ and LHe. The experimental results for $LN_2$ shows that the total heat inleak into inner vessel consists of 14% radiation and 86% conduction through the filling tube. Therefore, it is expected that the conduction heat in leak of the vessel for high $C_p/h_{fg}$ cryogenic fluids can be significantly reduced. powders. The amount of copper coating was 20wt%. In order to examine corrosion behavior of the electrodes, the corrosion current and the current density, in 6M KOH aqueous solution after removal of oxygen in the solution, were measured by potentiodynamic and cyclic voltamo methods. The results showed that Co in the alloy increased corrosion resistance of the electrode whereas Ni decreased the stability of the electrode during the charge-discharge cycles. The electrode used Si sealant as a binder showed a lower corrosion current density than the electrode used PTFE and the electrode used Cu-coated alloy powders showed the best corrosion resistance.

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Comparison with SAR Patterns of Biological Objects Contacted with Coaxial Waveguide Antenna Using MUR and GPML ABCs in the FDTD Method (유한차분법에서 MUR과 GPML 흡수경계조건을 이용한 동축 도파관 안테나에 접촉된 생체의 SAR 패턴 비교)

  • 구성모;권광희;이창원;원철호;조진호
    • The Journal of Korean Institute of Electromagnetic Engineering and Science
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    • v.9 no.2
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    • pp.149-158
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    • 1998
  • The SAR patterns of biological objects contacted with coaxial waveguide antennal has been investigated, in which the biological object was modeled by a homogeneous and four-layered lossy human body. We derived the finite-difference time-domain(FDTD) algorithm and equation of MUR and generalized perfectly matched layer(GPML) ABCs in cylindrical coordination. The coupling between coaxial waveguide antenna and a biological object was analyzed by use of MUR and GPML ABCs in the FDTD method to obtain the absorbed power patterns in the media. The specific absorption rates (SAR) distribution which was corresponding to the temperature distribution was calculated in each region by use of the steady-state response in the FDTD method. The SAR patterns of the FDTD method using MUR absorbing boundary conditions(ABCs) was compared with those of the FDTD method using GPML ABCs. The comparison exhibits that the penetration depth of the SAR patterns using MUR ABCs is deeper than that of the SAR patterns using GPML ABCs because of loss in free space. However, the spread in the lateral directions of the SAR patterns using GPML ABCs is smaller than of the SAR patterns using MUR ABCs.

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