• Title/Summary/Keyword: Membrane filtration

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Protective Effects of Chongmyunggongjin-dan on H2O2-induced C6 Glial Cell Death (H2O2로 유발된 C6 신경교세포 사멸에 대한 총명공진단의 보호 효과)

  • Hwang, Gyu-sang;Shin, Yong-jeen
    • The Journal of Internal Korean Medicine
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    • v.41 no.1
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    • pp.44-58
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    • 2020
  • Objectives: This study was conducted to identify the protective effects of Chongmyunggongjin-dan (CMGJD) on Hydrogen peroxide (H2O2)-induced apoptosis mechanisms in C6 glial cells. Method: We used CMGJD after distilled water extraction, filtration, and lyophilization. The ROS scavenging effect was examined by fluorescence microscopy. Expression levels of proteins related to ROS generation were investigated by western blotting. Functional changes in organelles related to Reactive oxygen species (ROS) generation were investigated by immunoblotting and by verifying expression level of relevant enzymes. Results: The CMGJD extract protected the cells against H2O2-induced morphological changes and DNA fragmentation, inhibited the increase of Heme_oxygenase-1(HO-1) and the decrease in catalase, protected against the loss of mitochondrial membrane potential, inhibited disturbances of lysosomal function, and induced an increase in peroxisomes. Conclusion: CMGJD was confirmed to have a protective effect on H2O2-induced C6 glial cell death possibly by blocking the pathways causing damage to subcellular organelles, such as mitochondria, lysosomes, and peroxisomes. We assume that CMGJD will be effective for the prevention and treatment of ischemic stroke in a clinical environment.

Determination of Antibiotic Residues: I. Extraction and Clean-up Methods for Solid Samples_A Review (시료 중 잔류 항생제 분석 방법: I. 고상 시료 전처리 방법)

  • Kim, Chansik;Ryu, Hong-Duck;Chung, Eu Gene;Kim, Yongseok;Rhew, Doug Hee
    • Journal of Korean Society on Water Environment
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    • v.32 no.6
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    • pp.600-627
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    • 2016
  • Korea is one of the countries with a large veterinary antibiotics market, although antimicrobial resistance in bacteria is becoming a serious issue in many countries. The Korean government started to take interest in estimating the effects of livestock manure on rivers and agricultural soils and in monitoring of heavy metals, organic pollutants and antibiotics in the ambient water and soil. In this paper, pre-treatment methods to separate the selected antibiotics from solid samples were reviewed. It is essential to select an efficient and appropriate procedure for pre-treatment due to the high proportion of proteins and organics in biosolid samples. Pre-treatment consists of extraction followed by clean-up. Initially, homogenized samples were extracted by sonication, mechanical agitation or pressurized liquid extraction with methanol/acetonitrile/water mixture under acidic/basic conditions depending on the compound. However, aminoglycosides and colistin were extracted with 5% trichloroacetic acid and HCl, respectively. Since the ${\beta}-lactams$ are easily decomposed in acidic and basic conditions, they were extracted in neutral pH. Filtration with a membrane (pore size, $0.2{\mu}m$) or solid phase extraction with HLB and methanol, as eluents, was normally applied for the clean-up. At least, three different pre-treatment procedures should be adopted to screen all the selected antibiotics in solid samples.

Determination of Antibiotic Residues: II. Extraction and Clean-up Methods for Liquid Samples_A Review (시료 중 잔류 항생제 분석 방법: II. 액상 시료 전처리 방법)

  • Kim, Chansik;Ryu, Hong-Duck;Chung, Eu Gene;Kim, Yongseok;Rhew, Doug Hee
    • Journal of Korean Society on Water Environment
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    • v.32 no.6
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    • pp.628-648
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    • 2016
  • Increased attention has been paid to the presence of veterinary antibiotics in various environmental matrices due to their toxicological behavior in the ecosystem and development of antibiotic-resistant strains of pathogenic bacteria. In the this review, 37 target antimicrobials were selected based on annual sales of antibiotics for livestock in South Korea 2014. Also, extraction and clean-up methods for the determination of the antibiotic residues in liquid samples including water, milk, and honey were comprehensively reviewed in the literature. Solid-phase extraction (SPE) was commonly used as a pre-treatment method for the samples. Most of the analytes were extracted in acidic conditions (2.5~4.0) except for aminoglycosides, which were extracted in neutral conditions (7.0~8.0). ${\beta}-Lactams$ showed the highest recoveries in neutral pH due to their degradation characteristics in acidic media. Starta-X, Oasis HLB, and Oasis MCX were frequently applied as an SPE cartridge and Oasis HLB showed the highest recoveries for the majority of antibiotic classes. The homogenized honey and milk were extracted by mixing with acids for deproteinization. Solids and other interfering substances in the extract were eliminated by centrifugation followed by membrane filtration or SPE before injection into HPLC.

A Study on the Adsorption Characteristics of Phenol in the presence of Humic Acid Using Activated Carbon Fiber (섬유상활성탄소를 이용한 Humic Acid 공존시 페놀의 흡착특성에 관한 연구)

  • Tak, Seong-Jae;Seo, Seong-Wen;Kim, Seong-Sun;Kim, Jin-Man
    • Journal of Korean Society of Water and Wastewater
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    • v.14 no.1
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    • pp.54-61
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    • 2000
  • Recently, our circumstances are threatened by an accident that leakage of under ground storage tank and illegal dumping of synthetic organic compounds at chemical plants and many treatment methods, Activated carbon adsorption, Ozonization, Membrane filtration and Photocatalystic oxidation, are developed to remove such a synthetic organic compounds. And it has reported that Activated carbon adsorption have a great removal efficiency to nondegradable matters and organic compounds which have a high molecular weight. Comparing with other adsorbents, Activated carbon adsorption have a worse efficiency when ad desorption speed is low. Thus improved type of adsorbents was invented and one of those is Activated Carbon Filter. The purpose of this study was getting information about adsorption characteristic phenol which can be applied Activated Carbon Fiber and Granular Activated Carbon. In detail, With comparing removal characteristics of phenol in the presence Humic Acid using Activated Carbon Fiber(ACF) and Granular Activated. Carbon(GAC), it is to certify an effective application of Activated Carbon Fiber. At the range of this study, Batch test, Isotherm adsorption test and Factorial analysis, following conclusion were obtained from the results of this study. Batch test was carried to know time of adsorption equilibrium. In this study about time of adsorption equilibrium by ACF was faster than GAC's, for developed micropore of ACF. From the result of phenol adsorption test, High removal rate of adsorption is shown at pH 5. The result of lsotherm adsorption test, it has represented that the Freundlich's isotherm is most suitable one in others, that a ACF's adsorption capacity is more excellent than GAC's. Adsorption of phenol exiting humic acid is decreased getting raised humic acid concentration. Since ACF's micropore is developed at this time, an effect of high molecular humic acid is lower. Factorial analysis was carried to know about Main effect which was injection dosage of adsorbent in the range of this study.

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Expression, Purification, Crystallization and Preliminary X-Ray Crystallographic Analysis of CnrX from Cupriavidus metallidurans CH34

  • Kim, Kook-Han;Jung, Eun-Jung;Im, Ha-Na;Lelie, Daniel Van Der;Kim, Eunice Eun-Kyeong
    • Journal of Microbiology and Biotechnology
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    • v.18 no.1
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    • pp.43-47
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    • 2008
  • The nickel and cobalt resistance of Cupriavidus metallidurans CH34 is mediated by the CnrCBA efflux pump encoded by the cnrYHXCBAT metal resistance determinant. The products of the three genes cnrYXH transcriptionally regulate expression of cnr. CnrY and CnrX are membrane-bound proteins, probably functioning as anti-sigma factors, whereas CnrH is a cnr-specific extracytoplasmic functions (ECF) sigma factor. The periplasmic domain of CnrX (residues 29-148) was cloned as a N-terminal His-tagged protein, expressed in Escherichia coli, and purified using affinity chromatography and gel filtration. The molecular mass was estimated to be about 13.6kDa by size exclusion chromatography, corresponding to a monomer. The tetragonal bipyramid crystals were obtained by mixing an equal volume of protein in 50mM Tris-HCl, pH 7.5, 1% glycerol, 100mM NaCl, 1mM DTT, and the reservoir solution of 15% w/v PEG 2000, 100mM lithium chloride at 277K in 2-4 days using hanging drop vapor diffusion. The protein concentration was 24mg/ml. The crystal that diffracted to $2.42{\AA}$ resolution belongs to space group $P4_1\;or\;P4_3$ with unit cell parameters of $a=b=32.14{\AA},\;c=195.31{\AA},\;{\alpha}={\beta}={\gamma}=90^{\circ}$, with one molecule of CnrX in the asymmetric unit.

Modification of Medium to Examine Fecal Coliforms in Water (물에서의 분원성대장균군 검사를 위한 최적 배지 선정)

  • Lee, Eunsook;Ko, Nayun;Choi, Byungdo;Kim, Bogsoon
    • Journal of Korean Society on Water Environment
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    • v.36 no.4
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    • pp.275-283
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    • 2020
  • Fecal coliforms are indicator bacteria to evaluate fecal contamination and microbiological safety in environment water. To examine fecal coliforms by membrane filtration, 1% rosolic acid solution dissolved in sodium hydroxide(0.2 M) should be added to m-FC medium according to Korean standard method. To reduce the exposure of researchers to harmful chemicals and expenditure of unnecessary cost, we evaluated if the rosolic acid solution is required to detect fecal coliforms. For 113 samples collected from five intake sources of Seoul, 42 samples of six tributaries, and 11 samples of sewage, the number of fecal coliforms was compared in medium with or without the reagent. As a result, the number was higher in m-FC medium without the reagent, but there was not a statistically significant difference. In the water intake, m-FC medium without the reagent could be used to examine fecal coliforms except in July, August and in case of rainfall. When heterotrophic plate counts exceeded 1,000 CFU/filter, or during rainfall, there was an effect of background bacteria in two types of the medium. However, it was more appropriate to use m-FC medium with the reagent to suppress gram-positive bacteria that can grow on medium without the reagent. In the tributary and sewage samples, the effect of the background bacteria was low, allowing the use of medium without the reagent regardless rainfall. Thus, it is necessary to present in standard method that the addition of rosolic acid solution in m-FC medium can be selected according to the characteristics of samples.

A Simple Method for the Concentration of Fish Pathogenic Virus in Sea Water (한외여과막을 이용한 해수내 어류 병원바이러스 농축법)

  • Oh, Myung-Joo;Kim, Suk-Ryul;Jung, Sung-Ju;Kim, Hyeung-Rak;Kim, Heung-Yun;Yeo, In-Kyu
    • Journal of fish pathology
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    • v.13 no.1
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    • pp.61-66
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    • 2000
  • A method was developed for concentrating fish pathogenic virus from sea water using membrane ultrafiltration system and centricon. The method consists of passing large volumes (Ca. 20 liter) of sea water through ultrafiltration (PAN) filter followed by cross-flow filtration method and centrifugation use the centricon (Plus-20). This procedure permitted the processing of 20 liter of sea water which resulted in a 20,000-fold reduction in the volume of water and greater than 90% recovery of the seeded MABV.

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Recovery of Pd(II), Pt(IV), and Rh(III) Using Polyelectrolytes

  • Lee, You-Sean;Lee, Hoosung;Chung, Koo-Soon
    • Analytical Science and Technology
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    • v.8 no.4
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    • pp.561-568
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    • 1995
  • Two methods, precipitation and ultrafiltration, were applied in order to recover platinum group metals(PGM) by complexing them with water-soluble polyelectrolytes, e.g., polyethyleneimine [PEl], poly(2-vinylpyridine) [2-PVP], poly (4-vinylpyridine) [4-PVP], and poly (styrene sulfonic acid) [PSSA]. In the precipitation method, the PGM-polyelectrolyte complex that was formed by mixing first with polybase, e.g.,4-PVP at pH 1 was precipitated by further mixing with polyacid, e.g., PSSA. However, the recovery of PGM obtained by this method was not quantitative(less than 70%). The "sandwiching" binding between the metal anions and two polyelectrolytes was examined by X-ray photoelectron spectroscopy(XPS). The XPS studies indicated that the PGM atom was bound with the acdic and basic polyelectrolyte via its oxygen and nitrogen atom, respectively. The recovery of PGM using polyelectrolyte was further studied by ultrafiltration methods as follows : The PGM ions, eomplexed at pH 1 with polyelectrolyte, allowed the applicntion of membrane filtration by virtue of the great differences in molecular weights between PGM and other low molecular weight species. By applying this method, Pd and Pt (ca. $10^{-4}M$) were selectively separated almost quantitatively from coexisting metal ions, e.g., $Cu^{2+}$ and $Ni^{2+}$. The EPR spectra and viscosity measurements indicated that these polyelectrlytes were not bound to $Cu^{2+}$ and $Ni^{2+}$ ions at this pH, which provided the basis for selective separation of PGM(Pd, Pt and Rh) from these coexisting ions.

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A Novel Simple Method to Purify Recombinant Soluble Human Complement Receptor Type 1 (sCR 1) from CHO Cell Culture

  • Wang, Pi-Chao;Hisamune Kato;Takehiro Inoue;Masatoshi Matsumura;Noriyuki Ishii;Yoshinobu Murakami;Tsukasa Seya
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.2
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    • pp.67-75
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    • 2002
  • The human complement receptor type 1 (CR 1, C3 b/C4b receptor) is a polymorphic membrane glycoprotein expressed on human erythrocytes, peripheral leukocytes, plasma and renal glomerular podocytes, which consists of transmembrane and cytoplasmic domains with 30 repeating homologous protein domains known as short consensus repeats (SCR). CR1 has been used as an inhibitor for inflammatory and immune system for the past several years. Recently; it is reported that CRl was found to suppress the hyper-acute rejection in xeno-transplantation and can be used to cure autoimmune diseases. A soluble form of CRl, called sCRl, is a recombinant CRl by cleaving the transmembrane domain at C-terminus and has been expressed in Chinese Hamster Ovary (CHO) cells. Several purification methods for sCR1 from CHO cells have been reported, but most of them require complicated steps at high cost. Moreover, such methods are mostly performed under the pH condition apt to denaturing sCR1 and causes sCRl losing its activity. We here report a rapid and efficient method to purify sCR1 from CHO cell. The new method consists of a two-stage of cell culture by cultivating cells in serum medium followed by serum-free medium, and a two-stage of column purification by means of heparin and gel filtration column chromatography. By using this novel method, sCR1 can be purified in a simple and effective way with high yield and purity, furthermore, the purified sCR1 was confirmed to retain its activity to suppress the complement activation in vivo and ex vivo.

Antithrombin Active Polysaccharide Isolated from the Alkaline Extract of Red Ginseng

  • Kim Dong Chung;In Man-Jin;Lee Ji Young;Hwang Yoon Kyung;Lee Sung Dong
    • Journal of Ginseng Research
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    • v.23 no.4
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    • pp.217-221
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    • 1999
  • We have isolated an antithrombin active polysaccharide in red ginseng by procedures comprising three major steps involving alkaline extraction, anion exchange and gel permeation chromatography. Active polysaccharide behaved as a single band on cellulose acetate membrane electrophoresis. The average molecular mass was estimated to be about 177 kDa by gel filtration. This polysaccharide was found to be an acidic heteropolysaccharide that contains uronic acid moiety $(40.2\%)$, sulfate group $(9.2\%)$ and protein $(1.5\%)$ in addition to neutral sugar consisted of rhamnose, mannose, galactose, arabinose, glucose, fucose and xylose in a molar ratio of 1.00 : 0.88 : 0.86 : 0.78: 0.70 : 0.33 : 0.22. This polysaccharide inhibited blood coagulation via the intrinsic pathway like heparin in a dose-dependent manner. The clotting of fibrinogen by thrombin was also mitigated by the presence of this polysaccharide.

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