• Title/Summary/Keyword: Ion gel

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Study on the Anti-HT-29 Human Colon Cancer Activity of $\beta$-Glucans and Their Enzymatically Hydrolyzed Oligosaccharides from Agalicus blazei Murill (아가리쿠스로부터 분리한 $\beta$-glucan과 그 올리고당류의 HT-29 인체 대장암 세포에 대한 항암 활성에 관한 연구)

  • Lee, Mi-Young;Kim, Ki-Hoon;Kim, Yea-Woon;Chang, Hun-Gil;Lee, Dong-Seok
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.319-325
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    • 2006
  • [ $\beta$ ]-Glucans (AG) were prepared from Agaricus blazei cultured in the medium fortified with the roots of Pueraria spp. by repeated extraction with hot water, gel filtration chromatography and DEAE ion exchange chromatography. Oligosaccharides (AO) were derived from the hydrolysis of AG by an endo-$\beta$-(1$\rightarrow$6)-glucanase from Bacillus megaterium. The anti-HT-29 human colon cancer activity of AG or AO was investigated using MTT assay, apoptosis assay, cell cycle analysis, and cDNA microairay. AG and AO both inhibited proliferation and growth of HT-29 cells, and stimulated apoptosis of the cells in a dose-dependent manner. In cell cycle analysis, treating HT-29 cells with AG or AO resulted in the increase of cells in the G0 (sub-G1) and G1 phase. Especially, AO was more effective in inducing G0/G1 cell cycle arrest than AG. To screen the genes involved in the increase of apoptosis, the gene expression profile of the HT-29 cells treated with AO was examined by cDNA microarray. While several genes involved in cell cycle progression (CCND2 and CDK2) were down-regulated, many genes involved in apoptosis (TNFSF9, TNFRSF9, FADD, CASP8, BAD, CRADD, CASP9 etc), cell cycle inhibitor (CDKN2A), immune response (IL6, IL18, IL6R etc), and tumor suppressor (CEACAM1, TP53BP2, IRF1, and PHB) were up-regulated. These results suggest that AO could inhibit the proliferation and growth of HT-29 cells by G0/G1 cell cycle arrest and induction of apoptosis.

Characteristic Properties of Fucoidan Sulfate Purified from Gompi, Ecklonia stolonifera (곰피에서 정제한 Fucoidan Sulfate의 특성)

  • Lee, Hong-Soo;Jin, Sung-Hyun;Kim, Hee-Sook;Ryu, Byung-Ho
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.716-723
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    • 1995
  • The fucoidan purified from Korean brown seaweed, Ecklonia stolonifera was characterized on molecular structure and blood anticoagulant activities. Extraction was conducted at $100^{\circ}C$ with water and repeated twice. The crude fucodian was 151.1g out of 20.0 kg of Ecklonia stolonifera. The Fucoidan-1, which was purified from crude fucoidan using calcium chloride and cetyl pyridium chloride (CPC), was 35.2% against crude fucoidan. Fucoidan-5 was obtained approximately 28.1% from Fucoidan-1 through DEAE-Toyopearl 650 M ion-exchange column chromatography and showed one band by cellulose acetate electrophoresis. The molecular weight of Fucoidan-5 was estimated to be about 21,000∼23,000 dalton by Sephacryl S-300 gel filtration chromatography. Fucoidan-5 consists of 35.7% of fucose and 4.3% of galactose and the molar ratio of fucose and sulfate was about one to one. IR spectrum of Fucoidan-5 showed absorption at $1240\;cm^{-1}\;and\;850\;cm^{-1}$ and specific rotation value, $[\alpha]$, was $[\alpha]$. These results suggests that the sulfate maybe bind at $C_{4}$ carbon on ${\alpha}-L-fucose$. Gas chromatograph of methyl alditol acetate revealed that Fucoidan-5 is a fucose containing sulfated polysaccharide with $({\alpha}l-2)\;or\;({\alpha}l-2)$ glycosidic linkage. Anti-thrombin activity of the Fucoidan-5 was estimated as 1.4 time stronger than heparin. From above results, the purification methods using CPC and ion exchange chromatography is effective tools for obtaining highly purified fucoidan from Gompi, Ecklonia stolonifera.

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Purification and Characterization of a Lipolytic Enzyme Produced by Aeromonas hydrophila PL43 (Aeromonas hydrophila PL43이 생산하는 지질분해 효소의 정제 및 특성)

  • Kim, Yong-Woo;Hong, Sung Wook;Chung, Kun Sub
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.130-139
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    • 2016
  • A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co2+, Cu2+, and Fe2+, but was recovered by metal chelating of EDTA. The Km and Vmax values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.

Analysis of Bacterials Community Structure in Leadchate-Contaminated Groundwater using Denaturing Gradient Gel Electrophoresis (Denaturing Gradient Gel Electrophoresis를 이용한 매립지 침출수로 오염된 지하수의 세균 군집 분석)

  • Kim Jai-Soo;Kim Ji-Young;Koo So-Yeon;Ko Kyung-Seok;Lee Sang-Don;Cho Kyung-Suk;Koh Dong-Chan
    • Microbiology and Biotechnology Letters
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    • v.34 no.2
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    • pp.166-173
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    • 2006
  • This research has been performed to clarify the relationship between hydrogeochemistry and bacterial community structure in groundwater contaminated with landfill leachate. We collected and analyzed samples from 5 sites such as leachate (KSG1-12), treated leachate (KSG1-16), two contaminated groundwaters (KSG1-07 and KSG1-08) and non-contaminated groundwater (KSG1-13). pH was 8.83, 8.04, 6.87, 6.87 and 6.53 in order; redox potential (Eh) 108, 202, 47, 200 and 154 mV; electric conductivity (EC) 3710, 894, 1223, 559 and 169.9 $\mu$S/cm; suspended solids (SS) 86.45, 13.74, 4.18, 0.24 and 11.91 mg/L. In KSG01-12, the ion concentrations were higher especially in $Cl^-$ and $HCO_3^-$ than other sites. The concentrations of Fe, Mn and $SO_4^{2-}$ were higher In KSG1-07 than in KSG1-08, and vise versa in $NO_3^{2-}$. In the comparison of DGGE fingerprint patterns, the similarity was highest between KSG1-13 and KSG1-16 (57.2%), probably due to common properties like low or none contaminant concentrations. Otherwise KSG1-08 showed lowest similarities with KSG1-13 (25.8%) and KSG1-12 (27.6%), maybe because of the degree of contamination. The most dominant bacterial species in each site were involved in $\alpha$-Proteobacteria (55.6%) in KSG1-12, $\gamma$-Proteobacteria (50.0%) in KSG1-16, $\beta$-Proteobacteria (66.7%) in KSG1-07, $\gamma$-Proteobacteria (54.5%) in KSG1-08 and $\beta$-Proteobacteria (36.4%) in KSG1-13. These results indicate that the microbial community structure might be changed according to the flow of leachate in grounderwater, implying changes in concentrations of pollutants, available electron accepters and/or other environmental conditions.

Structural and optical properties of Ni-substituted spinel $LiMn_2O_4$ thin films (니켈 치환된 스피넬 LiMn2O4 박막의 구조적, 광학적 성질)

  • Lee, Jung-Han;Kim, Kwang-Joo
    • Journal of the Korean Vacuum Society
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    • v.15 no.5
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    • pp.527-533
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    • 2006
  • Spinel $LiNi_xMn_{2-x}O_4$ thin films were synthesized up to x = 0.9 by a sol-gel method employing spin-coating. The Ni-substituted films were found to maintain cubic structure at low x but to exhibit tetragonal structure for $x{\geq}0.6$. Such cubic-tetragonal phase transition indicates that $Ni^{3+}(d7)$ ions with low-spin $(t_{2g}^6,e_g^1)$ state occupy the octahedral sites of the compound, thus being subject to the Jahn-Teller distortion. By x-ray photoelectron spectroscopy both $Ni^{2+}$ and $Ni^{3+}$ ions were detected. Optical properties of the $LiNi_xMn_{2-x}O_4$ films were investigated by spectroscopic ellipsometry (SE) in the visible?ultraviolet range. The measured dielectric function spectra by SE mainly consist of broad absorption structures attributed to charge-transfer (CT) transitions, $O^{2-}(2p){\rightarrow}Mn^{4+}(3d)$ for 1.9 $(t_{2g})$ and $2.8{\sim}3.0$ eV $(e_g)$ structures and $O^{2-}(2p){\rightarrow}Mn^{3+}(3d)$ for 2.3 $(t_{2g})$ and $3.4{\sim}3.6$ eV $(e_g)$ structures. Also, sharp absorption structures were observed at about 1.6, 1.7, and 1.9 eV, interpreted as due to d-d crystal-field transitions within the octahedral $Mn^{3+}$ ion. The strengths of these absorption structures are reduced by the Ni substitution. Rapid reduction of the CT transition strength involving the eg states for x = 0.6 is attributed to the reduced wavefunction overlap between the $e_g$ and the $O^{2-}(2p)$ states due to the tetragonal extension of the lattice constant by the Jahn-Teller effect.

Purification and Characterization of the Red Carotenoprotein from the Muscle of Blue Mussel, Mytilus edulis (진주담치 근육중의 적색 Carotenoprotein의 정제 및 특성)

  • YANG Huyn-Pil;LEE An-Jong;KIM Yong-Tae;KIM Se-Kwon
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.482-494
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    • 1994
  • Most of carotenoprotein complexes have been extracted by using buffered solutions. However, in this study carotenoprotein from the muscle of Blue mussel(Mytilus edulis) was extracted by a detergent such as Triton X-100. It was purified and characterized by $20\%$ (w/v) $(NH_4)_2SO_4$, DEAE-cellulose ion exchange and Sephacryl S-300 gel filtration. The carotenoprotein(${\lambda}_{max}=462nm$) had an approximate M. W. of 372KDa(gel filtration). SDS-PAGE analysis of the carotenoprotein indicated the presence of four polypeptides of 60KDa($23.70\%$), 46.9KDa($9.14\%$), 26KDa($49.14\%$) and 13KDa($18.02\%$). Carotenoprotein denaturated by treatment with SDS to a final concentration of $0.2\%$ (w/v) caused a hypsochromic shift of ${\lambda}_{max}$ from 462nm to 456nm. The carotenoprotein contained lipids as structure units. The amino acid composition of the carotenoprotein contained large essential amino acid amounts of $62.8\%$, and the content of threonine($35.9\%$) was higher than other amino acids, but histidine, methionine and proline were not present. In the carotenoprotein, the major fatty acids were $C_{16:4},\;C_{16:0},\;C_{20:5}\;and\;C_{22:6}$. The percentages of polyunsaturated fatty acids($62.4\%$) were higher compared to other fatty acids(saturated fatty acids $19.6\%$, monounsaturated fatty acids $18.0\%$). Carotenoid was extracted from the carotenoprotein by acetone and it was separated into five different components by preparative TLC(benzene:petroleum ether:acetone=69:17:14). The major components of carotenoid were mytiloxanthin($74.79\%$) and 3,4,3'- trihydroxy-7',8'-didehydro-${\beta}$-carotene($18.26\%$), and they were at least presented as prosthetic groups of carotenoprotein.

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Purification of antigenic proteins of Paragonimus westermani and their applicability to experimental cat paragonimiasis (폐(肺)디스토마(Paragonimus westermani) 감염(感染) 고양이 혈청(血淸)에 대(對)한 ELISA 항체가(抗體價)의 의의(意義))

  • Choi, Won-Young;Yoo, Jae-Eul;Nam, Ho-Woo;Choi, Hyung-Rak
    • Parasites, Hosts and Diseases
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    • v.24 no.2
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    • pp.177-186
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    • 1986
  • This study was designed to evaluate the partially purified antigens which were fractionated from crude extract of Paragonimus westermani and to monitor the enzyme-linked immunosorbent assay (ELISA) in experimental cat paragonimiasis during the course of infection as well as before and after chemotherapy. Crude extract of 6-month-old adult P. westermani was fractionated to 5 antigens by successive applications of ammonium sulfate precipitation, ion exchange chromatography and gel filtration. And the cats, 10 in each group, were infected with 60, 30, 15, and 5 metacercariae, then the half of each group was treated with praziquantel 2 times in one day of 100mg per kilogram of weight on 150 days after the infection. Sera were collected every 10 days. ELISA was performed with the concentration of $2{\mu}g/ml$ antigen, 100 times diluted sera and 1,000 times diluted alkaline phosphatase conjugated anti-cat IgG. The results were as follows: 1. Absorbance by ELISA with proteins precipitated by differential concentration of ammonium sulfate was the highest at $51{\sim}65%$ precipitate (PA2), followed by $0{\sim}50%$ precipitate (PAl), $66{\sim}80%$ precipitate (PA3), and $81{\sim}90%$ precipitate (PA4). Unprecipitated protein over 90% ammonium sulfate (PA5) showed the lowest antigenicity. 2. Fractionation of PA1, PA2, and PA3 through the DEAE-cellulose column did not differentiate the antigenic proteins. 3. By passing through the Sephadex G-200 column, PA1 and PA2 were fractionated to high molecular weight proteins and those of low molecular weight which showed high absorbance by ELISA (PA1-I, II and PA2-I, II). But PA3 was shown to have a fraction of high molecular weight proteins (PA3-I) which showed high antigenicity. 4. SDS-polyacrylamide gel electrophoresis of PA1-I, P A1-II, PA2-I, PA2-II, PA3-I, and crude extract was performed. Fraction PA1-I was composed of proteins which had the molecular weight of 270 kilodaltons(KD) to 196 KD; of them 220KD protein was major band. Fraction PA2-I was composed of $255{\sim}225\;KD$, and PA3-I, $255{\sim}240\;KD$, respectively. Fraction PA1-II and fraction PA2-II consisted of 30 KD proteins. 5. Absorbance by ELISA began to increase within $10{\sim}20$ days after the infection and reached the highest on $140{\sim}180$ days, then made plateau thereafter. 6. Absorbance by ELISA decreased after praziquantel treatment. In 60 metacercariae infection group, the absorbance had been decreasing, but remained within the positive range during observation period, while those of 30, 15, and 5 metacercariae infection groups turned to negative range. 7. Fraction PA1-II showed the highest antigenicity in ELISA, then fraction PA2-I, fraction PA1-I, fraction PA2-II, fraction PA3-I and crude extract followed. In early phase of infection, the absorbance of fraction PA1-II showed more rapid increase than those of the other fractions and it came to positive range at $20{\sim}30$ days after infection.

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Performance Evaluation of Backwash Hydrodynamic Separator Filter for Treatment of Micro Particles (역세척 Hydrodynamic Separator Filter를 이용한 미세입자 제거 특성 분석)

  • Lee, Jun-Ho;Bang, Ki-Woong
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.10
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    • pp.694-701
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    • 2012
  • The main purpose of this study is to evaluate of backwash system of hydrodynamic separator filter (HSF) with solar powered submerged pumps. It consists of a photovoltaic solar array, control electronics, battery, and two submersible pump powered by a 12 voltage DC motor. The laboratory scale study on treatable potential of micro particles using backwash HSF that was a combined with perlite filter cartridge and backwash nozzles. Since it was not easy to use actual storm water in the scaled-down hydraulic model investigations, it was necessary to reproduce ranges of particle sizes with synthetic materials. The synthesized storm runoff was made with water and addition of particles; ion exchange resin partices, silica gel particles, and commercial area manhole sediment particles. HSF was made of acryl resin with 250 mm of diameter filter chamber and overall height of 800 mm. Four case test were performed with different backwashing conditions and determined the SS removal efficiency with various surface loading rates. The operated range of surface loading rate was about 308~$1,250m^3/m^2/day$. It was found that SS removal efficiency of HSF using two submersible pumps improved by about 18% compared with HSF without backwash. Nonpoint control devices with solar water pumping systems would be useful for backwashing the filter in areas with not suppling electricity and reduce filter media exchange cost.

Tungsten-Doped Titania Nanopowders - Their Chemical Vapor Synthesis and Photocatalytic Activity (텅스텐이 도핑된 티타니아 나노분말의 화학기상합성 및 광촉매 활성)

  • Park, Bo-In;Kang, Kae-Myung;Jie, Hyunseock;Song, Bong-Geun;Park, Jong-Ku;Cho, So-Hye
    • Journal of the Korean Institute of Gas
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    • v.16 no.6
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    • pp.143-147
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    • 2012
  • Photocatalytic properties of $TiO_2$ nanopowders has been received much attention due to their high potentials for environmental applications such as remediation of polluted environments. The $TiO_2$ nanopowders doped with metal or non-metal elements have been synthesized by variety methods such as flame method, chemical vapor synthesis, sol-gel, ion implantation, which affect a doping behavior in different ways resulting in different surface characteristics, leading to different photocatalytic activity. In addition to an effect of synthesis methods, the photocatalytic activity of $TiO_2$ nanopowders can be improved by subsequent heat-treatments. In this study, to obtain a highly efficient photocatalyst, we synthesized $TiO_2$ nanopowders doped with tungsten by the chemical vapor synthesis method (CVS) and determined their physical properties and photocatalytic activity, together with subsequent post-treatment in the range of $300^{\circ}C$ to $700^{\circ}C$.

Physicochemical Properties and Dietary Effect of Glycoprotein from Sea Cucumber(Stichopus japonicus) (해삼 당단백질의 물리화학적 특성과 식이효과)

  • 류홍수;문정혜;유병진;문수경
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.25 no.2
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    • pp.240-248
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    • 1996
  • To clarify the potentiality of sea cucumbers as dietary food, the effects of those glycoprotein on dietary proteins and physicochemical properties of those proteins were studied. Crude glycoprotein was efficiently extracted using 20mM sodium phosphate beffer(pH 7.0) and by salting out with 80% ammoniym sulfate saturation. The fractions obtained through the DEAE-cellulose ion exchange chromatography was identified as glycoprotein by Schiff's reagent and SDS polyacrylanide gel electro-phoresis. The yields of each glycoprotein from the three kinds of sea cucumbers were 0.814(red), 0.184(blue) and 0.232(black) and the molecular weights of the glycoproteins subunits were ranged from 20,000 dalton(blue and black) to 29,000 dalton(red), respectively. The electrophoretic patterns of the glycoprotein isolates were similar to each other and any significant difference in amino acid pattern was observed. Predominant arnino scids were Asx(aspartic acid and asparagine) and Glx(glutamic acid and glutamine) ; in contrast, histidine and methionine were below 2% as compared to total amino acids. water holding capacities of the glycoprotein isolates from red, blue and black cucumbers were equally 100% and emulsion activities ranged from 53% to 64%. In addition the emulsion stabilities were 7.04, 1.37 and 2.44, respectively. In vitro digestibility of some proteins(casein, SPI and squid) was decreased as increasing the level of the freeze dried sea cucumber powder and glycoprotein isolates. But squid protein was not affected.

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