• Title/Summary/Keyword: 컬럼

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A Nucleic Acid Amplification Tests for Reliable HCV RNA Detection Method for Plasma-Derived Products (핵산증폭시험을 이용한 혈장분획물질에서 HCV RNA 검출)

  • Hong, Seung-Hee
    • Korean Journal of Microbiology
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    • v.44 no.4
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    • pp.293-298
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    • 2008
  • HCV is transmitted via various plasma derived products. Current methods to detect hepatitis C virus (HCV) are based on its antibody detection in the donated blood and plasma. Viral contamination can potentially escape such detection during the window period of infection, when no antibody is present or the level of antibody is too low to detect. It is trying to application of nucleic acid amplification tests (NAT) for the direct detection of HCV. The objective of this study was to develop a reliable NAT for the HCV RNA detection from plasma-derived products. The most useful primers was selected for NAT among 5 sets of primers. We have also found that QIAamp viral RNA isolation kit was the most efficient for HCV RNA isolation. The highest sensitivity and specificity was appeared in $48^{\circ}C$ annealing temperature and 30 pmol of primers. With a spiking of HCV to albumin, immunoglobulins and coagulation factors, NAT can detect up to 100 IU/ml. Meanwhile, COBAS amplicor HCV 2.0 afforded a lower sensitivity in high concentrated intramuscular immunoglobulins to below 500 IU/ml. Our results suggested that NAT appears to be a highly sensitive and specific method for HCV RNA detection in plasma-derived products.

Experimental and FEMLAB Simulation Study of Ibuprofen Racemate Separation in HPLC (Ibuprofen Racemate의 HPLC 분리실험과 FEMLAB 전산모사 연구)

  • Lee, Eun;Chang, Sang-Mork;Kim, Jong-Min;Kim, Woo-Shick;Kim, In-Ho
    • KSBB Journal
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    • v.21 no.3
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    • pp.224-229
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    • 2006
  • FEMLAB is a powerful interactive environment for modeling, solving all kinds of scientific and engineering problems based on partial differential equations(PDEs). Separation process of chiral compound in HPLC columns was simulated by FEMLAB. To study change of elution profile with isotherm models, non-competitive and competitive Langmuir adsorption isotherm were adopted. Separated material was (R, S)-ibuprofen [(R, S)-2-(4-isobutyl phenyl) propionic acid], an anti-inflammatory agent, which retain the pharmacological activity in the (S)-(+)-enantiomer. Sample concentrations were changed from 0.5 mg/ml to 2.0 mg/ml at a flow rate of 1 ml/min and flow rate varied from 1 ml/min to 3 ml/min at an ibuprofen concentration of 2.0 mg/ml and $20{\mu}l$ of injection volume. Simulated results were well fitted with experimental data.

Characterization and Cloning of a Phytase from Escherichia coli WC7. (Escherichia coli WC7가 생산하는 Phytase의 효소특성과 그 유전자의 클로닝)

  • 최원찬;오병철;김형권;강선철;오태광
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.1-7
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    • 2002
  • Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

Analysis of Flumeqnine in Meats (식육 중 항생제 flumequine의 분석)

  • Kuk, Ju-Hee;Ko, Yong-Seok;Kim, Yong-Hoon;Yoon, Chang-Yong;Sun, Nam-Kyu;Kim, Eun-Jung;Seo, Jee-Woo;Park, Ji-Won;Kang, Kil-Jin
    • Korean Journal of Food Science and Technology
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    • v.41 no.1
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    • pp.7-10
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    • 2009
  • An analytical method for the determination of flumequine in meats was developed and validated using high-performance liquid chromatography with fluorescence detection. The samples were mixed with sodium sulfate and extracted with ethyl acetate. After clean-up, the residues were dissolved in mobile phase. The calibration curves showed high linearity ($r^2$=0.9979) within the concentration range of 0.1-1.0 mg/kg. The limit of detection and limit of quantification were validated at 0.005 and 0.017 mg/kg, respectively. The recoveries in fortified meats ranged from 90.8 to 101.1%. The method was then validated in correspondence with the CODEX guidelines for flumequine residue in meats. Herein we monitored 150 samples of meats that were purchased in Korea (Seoul, Busan, Daegu, Daejeon, and Gwangju). Among the tested samples, flumequine was detected in 1 of beef and 1 of pork at levels in the range of 0.048-0.080 mg/kg. Overall, the flumequine residues in the tested samples were within the Maximun residue limit.

Risk Assessment and Safety Measures for Methanol Separation Process in BPA Plant (BPA 공장의 메탄올 분리공정에서 위험성 평가 및 안전대책)

  • Woo, In-Sung;Lee, Joong-Hee;Lee, In-Bok;Chon, Young-Woo;Park, Hee-Chul;Hwang, Seong-Min;Kim, Tae-Ok
    • Journal of the Korean Institute of Gas
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    • v.16 no.3
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    • pp.22-28
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    • 2012
  • For a methanol separation column of the BPA (Bisphenol A) plant, HAZOP (hazard and operability) assessment was performed and damage ranges were predicted from the accident scenarios for the fire and the explosion. As a result, the damage range of the jet fire was 20 m in the case of rupture of the discharge pipe (50 mm diameter) of safety valve, and that of the flash fire was 267 m in the case of catastrophic rupture. Also, the damage ranges of the unconfined vapor cloud explosion (UVCE) for the rupture of the discharge pipe and for the catastrophic rupture were 22 m and 542 m, respectively. For the worst case of release scenarios, safety measures were suggested as follows: the pressure instruments, which can detect abnormal rise of the internal pressure in the methanol separation column, should be installed by the 2 out of 3 voting method in the top section of the column. Through the detection, the instruments should simultaneously shut down the control and the emergency shut-off valves.

Anticariogenic activity of acanthoic acid isolated from Acanthopanax koreanum $N_{AKAI}$ against oral pathogens (섬오가피로부터 분리된 acanthoic acid의 항치아우식효과)

  • Kim, Hong-Jun;Jeong, Seung-Il;Lee, Hwa-Jung;Ju, Young-Sung
    • Korean Journal of Oriental Medicine
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    • v.12 no.1
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    • pp.69-75
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    • 2006
  • This research was aimed to search for natural antimicrobial agents that are sefe for humans and specific for oral pathogens. Acanthoic was isolated from the chloroform fraction of methanol extract of Acanthopanax koreanum $N_{AKAI}$ and its structure were elucidated by 13-NMR, 1H-NNR and ESIMS. Antibacterial activity of acanthoic acid was investigated by the minimum inhibitory (MIC) and minimun bactericidal (MBC) concentration. MIC/MBC of acanthoic acid against Streptococcus mutans $N_{AKAI}$ causing dental caries was determined to be $2/4\;{\mu}g/mL$, which was much lower than these of other natural antimicrobial agents such as $8/16\;{\mu}g/mL$ of sangurinarine and $250/500\;{\mu}/mL$ of green tea extract, $500/600\;{\mu}g/mL$ of thymol and borneol. Acanthoic and significantly inhibited the growth of other cariogenic bacteria such as Streptococcus sobrinus $N_{AKAI}$ and Streptococcus sanguis $N_{AKAI}$, and Streptococcus gordonii $N_{AKAI}$ in the MIC range of $4{\sim}32\;{\mu}g/mL$. My finding suggests that acanthoic acid could be employed as a potential antibacterial agent for preventing dental caries.

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Catalytic CVD-Kinetics of Pyrolytic Carbon and SiC on the Stainless Steel Stent (Stainless Steel Stent에 Pyrolytic Carbon과 SiC의 촉매적 CVD-Kinetic연구)

  • 이보성;이무용
    • Proceedings of the KAIS Fall Conference
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    • 2000.10a
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    • pp.30-33
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    • 2000
  • 최근 국내에서도 관 동맥 질환 환자의 수가 급증하고 있으며, 관 동맥 질환의 치료 방법인 관 동맥 성형 술은 관 동맥 stent의 도입에 의하여 보편화되어 국내에서 년간 5000개 이상의 stent가 시술되고 있다. 그러나 stent는 고가(1,200천원/개)로 전량 수입에 의존하고 있으며, 시술 후 사망까지 이를 수 있는 혈전에 의한 급성 페쇠와 재 협착이 문제점이다. 이를 위한 한가지 방법이 생체 적합성이 뛰어난 복합 stent의 개발인데 SiC나 Carbon을 coating한 stent는 시술 후 혈전 형성을 억제하는 것으로 알려져 있다. 특히 가장 순수한 Pyrolytic carbon은 hemocompatibility가 탁월하고 기밀 성이기 때문에 본 연구에서 그의 CVB-Kinetics를 연구코저 하는 것이다. methane으로부터 pyrolytic carbon의 CVD는 온도에 따라서 다양한 구조를 가지며 따라서 그의 mechanism도 다양하다는 것은 잘 알려져 있다. 더구나 광간(균질)반응과 표면(불균질)반응의 정량적 관계에 따라서도 다르다는 것도 확인되었다. 그러나 stainless steel 316L로 만든 stent는 12 - 15 %의 Ni과 2%의 Mo을 함유해서 금속성을 잃지 않는 저온(600℃)에서도 pyrolytic carbon의 속매적 CVD가 가능함을 그리고 SiC의 코팅에 적합한 buffer layer 역할을 함을 확인하였다. 그리하여 본 연구는 반응기 설계에 필요한 저온 촉매적 pyrolytic carbon의 CVD-kinetics의 연구결로 그의 mechanism과 함께 rate law 식을 유도, 확인하였으며 600℃, 90kPa에서 P/sub ch4//P/sub H2/=5:1과 체류시간 1.8 sec가 최적임을 발견하였다. 이때 석출속도 11.2 g-mol/g-cat.h 혹은 두께속도로 73 nm/sec를 나타내었다.메타놀-물 (1 : 1) 유출액에서 $(0.80\;{\mu}g)$ 검출되었다. 하면 morey eel내장에서 얻은 독물질도 DEAE-셀루로즈에서 ST-1 과 ST-2로 나누어지며, 이 ST-1의 TLC, HPLC 및 알루미나 컬럼상의 거동이 파랑비늘돔에서 얻은 ST-1의 그것과 같으므로 scaritoxin으로 보고한 ST-1은 ciguatoxin의 형태인 less polar cigutoxin (LPCTX) 으로 생각된다.에서 각각 대조구의 57, 413 및 315% 증진되었다. 거품의 열안정성은 15분 whipping시, pH 4.0(대조구, 30.2%) 및 5.0(대조구, 23.7%)에서 각각 $0{\sim}38.0$$0{\sim}57.0%$이었고 pH 7.0(대조구, 39.6%) 및 8.0(대조구, 43.6%)에서 각각 $0{\sim}59.4$$36.6{\sim}58.4%$이었으며 sodium alginate 첨가시가 가장 양호하였다. 전체적으로 보아 거품안정성이 높은 것은 열안정성도 높은 경향이며, 표면장력이 낮으면 거품형성능이 높아지고, 비점도가 높으면 거품안정성 및 열안정성이 높아지는 경향이 있었다.protocol.eractions between application agents that are developed using different languages. Dynamic agent invocation is accomplished by Java Native Interface(JNI) that links two heterogeneous methods, and by KQML language interface that facilitates the communications between heterogeneous agents. This scheme of dyna

Continuous Production of Authentic Human Growth Hormone from Methionyl Human Growth Hormone Using the Column Reactor of Immobilized Aminopeptidase M (고정화 Aminopeptidase M 컬럼 반응기를 이용한 메치오닐 인간성장호르몬으로부터 천연형 인간성장호르몬의 연속생산)

  • 이성희;김기태
    • KSBB Journal
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    • v.10 no.3
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    • pp.283-291
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    • 1995
  • The characteristics of aminopeptidase M(ApM) immobilized covalently on Cellufine Formyl and the continuous production of authentic human growth hormone(hGH) from methionyl human growth hormono(met-hGH) using the column reactor packed with immobilized ApM were investigated. Immobilized ApM with the proportion of 2.3mg ApM per 1g Cellufine Formyl gel had the highest met-hGH conversion activity. The optimum pH(7.0) and temperature($55^{\circ}C$) showed no appreciable difference between free and immobilized enzymes and the optimum temperature in continuous operation of the column reactor was also found to be $55^{\circ}C$. Under the conditions at which met-hGH was converted completely to hGH, the yield and productivity were about 77% and 0.8mg hGH/ml$.$h, respectively. In two column reactors of different sizes, met-hGH was converted to hGH with the same conversion rates and hGH yields at the same space velocities. The half-life of the reactor systems at $45^{\circ}C$ and $55^{\circ}C$ were projected from the continuous operations for 90 days to be 225 days and 81 days, respectively.

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Gene Cloning, Purification and Characterization of Xylanase 10A from Paenibacillus woosongensis in Escherichia coli (Paenibacillus woosongensis로부터 대장균에 Xylanase 10A의 유전자 클로닝과 정제 및 특성분석)

  • Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.48 no.2
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    • pp.158-166
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    • 2020
  • A gene coding for the xylanase was cloned from Paenibacillus woosongensis, followed by determination of its complete nucleotide sequence. This xylanase gene, designated as xyn10A, consists of 1,446 nucleotides encoding a polypeptide of 481 amino acid residues. Based on the deduced amino acid sequence, Xyn10A was identified to be a modular enzyme composed of a catalytic domain highly homologous to the glycosyl hydrolase family 10 xylanase and a putative carbohydrate-binding module (CBM) in the C-terminus. By using DEAE-sepharose and phenyl-sepharose column chromatography, Xyn10A was purified from the cellfree extract of recombinant Escherichia coli carrying a P. woosongensis xyn10A gene. The N-terminal amino acid sequence of the purified Xyn10A was identified to exactly match the sequence immediately following the signal peptide predicted by the Signal5.0 server. The purified Xyn10A was a truncated protein of 33 kDa, suggesting the deletion of CBM in the C-terminus by intracellular hydrolysis. The purified enzyme had an optimum pH and temperature of 6.0 and 55-60℃, respectively, with the kinetic parameters Vmax and Km of 298.8 U/mg and 2.47 mg/ml, respectively, for oat spelt xylan. The enzyme was more active on arabinoxylan than on oat spelt xylan and birchood xylan with low activity for p-nitrophenyl-β-xylopyranoside. Xylanase activity was significantly inhibited by 5 mM Cu2+, Mn2+, and SDS, and was noticeably enhanced by K+, Ni2+, and Ca2+. The enzyme could hydrolyze xylooligosaccharides larger than xylobiose. The predominant products resulting from xylooligosaccharide hydrolysis were xylobiose and xylose.

Removals of Formaldehyde by Silver Nano Particles Attached on the Surface of Activated Carbon (나노 은입자가 첨착된 활성탄의 포름알데히드 제거특성)

  • Shin, Seung-Kyu;Kang, Jeong-Hee;Song, Ji-Hyeon
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.10
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    • pp.936-941
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    • 2010
  • This study was conducted to investigate formaldehyde removals by silver nano-particles attached on the surface of granular activated carbon (Ag-AC) and to compare the results to those obtained with ordinary activated carbon (AC). The BET analysis showed that the overall surface area and the fraction of micropores (less than $20{\AA}$ diameter) of the Ag-AC were significantly decreased because the silver particles blocked the small pores on the surface of the Ag-AC. The formaldehyde removal capacity of the Ag-AC determined using the Freundlich isotherm was higher than that of AC. Despite the decreased BET surface area and micropore volume, the Ag-AC had the increased removal capacity for formaldehyde, presumably due to catalytic oxidation by silver nano-particles. In contrast, the adsorption intensity of the Ag-AC, estimated by 1/n in the Freundlich isotherm equation, was similar to that of the ordinary AC, indicating that the surface modification using silver nano-particles did not affect the adsorption characteristics of AC. In a column experiment, the Ag-AC also showed a longer breakthrough time than that of the AC. Simulation results using the homogeneous surface diffusion model (HSDM) were well fitted to the breakthrough curve of formaldehyde for the ordinary AC, but the predictions showed substantial deviations from the experimental data for the Ag-AC. The discrepancy was due to the catalytic oxidation of silver nano-particles that was not incorporated in the HSDM. Consequently, a new numerical model that takes the catalytic oxidation into accounts needs to be developed to predict the combined oxidation and adsorption process more accurately.