• Title/Summary/Keyword: Molecular Separation

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Analysis of Cyclic Adenosine Monophosphate (cAMP) Separation via RP-HPLC (reversed-phase high-performance liquid chromatography) by the Moment Method and the van Deemter Equation (역상 크로마토그래피에서 모멘트 방법과 van Deemter 식을 이용한 고리형 아데노신 일인산의 분리특성 연구)

  • Lee, Il Song;Ko, Kwan Young;Kim, In Ho
    • Korean Chemical Engineering Research
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    • v.53 no.6
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    • pp.723-729
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    • 2015
  • The moment analysis of cyclic adenosine monophosphate (cAMP) was performed using chromatograms that were obtained with the pulse input method from an octadecyl silica (ODS) high-performance liquid chromatography (HPLC) column. The general rate (GR) model was employed to calculate the first absolute moment and the second central moment. Three important coefficients for moment analysis, which are molecular diffusivity ($D_m$), external mass transfer coefficient ($k_f$), and intra-particle diffusivity ($D_e$), were estimated by the Wilke-Chang equation, Wilson-Geankoplis equation, and comparing van Deemter equation to theoretical plate number equation, respectively. Experiments were conducted by various conditions of flow rates, methanol volume ratio of the mobile phase, and solute concentration. After the moment analysis, results were organized by van Deemter plots. Also van Deemter coefficients were compared each other to effect $H_{ax}$, $H_f$, and $H_d$ on height equivalent to a theoretical plate (HETP, $H_{total}$). The value of intraparticle diffusion ($H_d$) was the primary factor which makes for HETP whereas external mass transfer ($H_f$) was disregardable factor.

Consecutive automated production of carbon-11 labeled radiopharmaceuticals by sharing 11C-methylation reagent from one 11C-synthetic module

  • Park, Hyun Sik;Lee, Hong Jin;An, Hyun Ho;Moon, Byung Seok;Lee, Byung Chul;Kim, Sang Eun
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.2 no.2
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    • pp.123-131
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    • 2016
  • Increasing clinical demand for carbon-11 labeled radiopharmaceuticals has triggered technological advances in fields of radiochemistry and automated modules. Even though carbon-11 has a short half-life ($t_{1/2}=20.4min$), the consecutive second production of carbon-11 labeled radiopharmaceutical in one $^{11}C$-synthetic module should be delayed at least over 4 h to avoid the high radiation exposure. We herein aimed to produce two different carbon-11 labeled radiopharmaceuticals ([$^{11}C$]PIB and [$^{11}C$]methionine) by sharing of [$^{11}C$]methylation source in one $^{11}C$-synthetic module. The synthesis of $^{11}C$-labeling reagents ($[^{11}C]CH_3I$ or $[^{11}C]CH_3OTf$) is fully automated using the commercial TRACERlab $FX_{C-pro}$ module and is readily adaptable to $^{11}C$-labeling reactor for [$^{11}C$]PIB as well as another $^{11}C$-labeling apparatus for [$^{11}C$]methionine via the three-way valve. After completing the [$^{11}C$]PIB production, the re-synthesized $[^{11}C]CH_3I$ was passed through the three-way valve connected the polyetheretherketone (PEEK) line and loaded into the C18 Sep-Pak cartridge including the methionine precursor. The labeled product [^${11}C$]methionine was purified by a simple cartridge separation and reformulated into saline. The radiochemical yield of [$^{11}C$]PIB and [$^{11}C$]methionine were $5.3{\pm}0.6%$ and $18.7{\pm}0.8%$ (n.d.c.), respectively, with over 97% of radiochemical purity. The specific activity of [$^{11}C$]PIB was over $110GBq/{\mu}mol$. Total production time of two radiopharmaceuticals needs about 2 h from $1^{st}$ beam irradiation including quality control tests. Final [$^{11}C$]PIB and [$^{11}C$]methionine were satisfied all quality control test standards.

Two-dimensional gel Electrophoresis of Helicobacter pylori for Proteomic Analysis

  • Jung, Tae-Sung;Kang, Seung-Chul;Choi, Yeo-Jeong;Jeon, Beong-Sam;Park, Jeong-Won;Jung, Sun-Ae;Song, Jae-Young;Choi, Sang-Haeng;Park, Seong-Gyu;Choe, Mi-Young;Lee, Byung-Sang;Byun, Eun-Young;Baik, Seung-Chul
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.2
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    • pp.97-108
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    • 2000
  • Two-dimensional gel electrophoresis (2-DE) is an essential tool of proteomics to analyse the entire set of proteins of an organism and its variation between organisms. Helicobacter pylori was tried to identify differences between strains. As the first step, whole H. pylori was lysed using high concentration urea contained lysis buffer [9.5 M Urea, 4% CHAPS, 35 mM Tris, 65 mM DTT, 0.01% SDS and 0.5% Ampholite (Bio-Rad, pH 3-10)]. The extract ($10\;{\mu}g$) was rehydrated to commercially available immobilised pH gradient (IPG) strips, then the proteins were separated according to their charges as the first dimensional separation. The IPG strips were placed on Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) to separate according to molecular mass of the proteins as the second dimension. The separated protein spots were visualised by silver staining in order to compare different expression of proteins between strains. Approximately 120 spots were identified in each mini-protein electrophoresised gel, furthermore about 65 to 75 spots were regarded as identical proteins in terms of pI value and molecular weight between strains used. In addition, distinct differences were found between strains, such as 219-1, Y7 and Y14, CH150. Two representative strains were examined using strips which had pH range from 4 to 7. This strips showed a number of isoforms which were considered large spots on pH range 3-10. Furthermore, the rest of spots on pH 4-7 IPG strips appeared very distinctive compared to broad range IPG strips. 2-DE seems to be an excellent tool for analysing and identifying variations between H. pylori strains.

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Evolution of sea Urchin Strongylocentrotus intermedius Based on DNA Sequences of a Mitochondrial Gene, Cytochrome c Oxidase Subunit I (미토콘드리아 유전자, 치토그롬 옥시다제(subunit I)의 염기서열을 이용한 새치성게(Strongylocentrotus intermedius)의 진화과정 분석)

  • Lee, Youn-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.5 no.2
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    • pp.157-168
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    • 2000
  • Sea urchin S. intermedius occurring in the Korean east coast is a cold water species that belongs to the family Strongylocentrotidae of Echinoidea. Although it is known that there are nine species in the family, species identification criteria, phylogenetic relationships, time and process of evolution of the family members have not been uncovered clearly. In the present study, I tried to find some clues to such problems for S. intermedius by means of DNA sequences. For this, cytochrome c oxidase subunit I (COI), one of the mitochondrial genes that evolve fast and follow maternal inheritance was analyzed. DNA was extracted from the female gonad of S. intermedius, a segment of COI gene amplified by polymerase chain reaction (PCR), and finally a total of 1077 base pair sequence of COI obtained by cloning and sequencing the PCR product. The sequence was compared with homologous genes of other sea urchins and echinoderm species. Phylogenetic trees of the COI gene segment revealed that S. intenedius is a sister species of S. purpuratus which lives along the east coast of the Paciflc. With reference to the fossil records of sea urchins and genetic distances in the molecular phylogenies, it is estimated that the two species were separated about 0.89 million years ago when the earth temperature fluctuated significantly. The current disjunct distribution patterns of the two species and the climate change of the earth at the time of separation suggest that speciation might have occurred by vicariance. The COI gene sequence obtained here now can be used as a molecular character which discerns S. intermedius from the other sea urchin species of Strongylocentrotidae.

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Effects of Fabrication Conditions on Electro-optic Properties of UV-Cured Polymer/Liquid Crystal Composite Films (UV 경화형 고분자/액정 복합체의 제조 조건에 따른 전기광학적 특성)

  • Park, Se Kwang;Park, Lee Soon;Keum, Chang Dae;Seok, Jae Wook;Ahn, Won Sool
    • Applied Chemistry for Engineering
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    • v.9 no.4
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    • pp.579-584
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    • 1998
  • Polymer dispersed liquid crystal(PDLC) composite films were made by polymerization induced phase separation method using UV-curing to investigate the effect of fabrication conditions, such as photoinitiator concentration, film thickness, polymerization temperature, and electric field during polymerization, etc., on the electro-optic properties. As the amount of photoinitiator increased, the driving voltage of PDLC device increased due to the increase of small-size liquid crystal phases. This was considered as the results from the increased interfacial area between liquid crystal (LC) and polymer matrix, since LC molecules at the interfacial regions were relatively difficult to response for the applied electric field. When the higher molecular weight oligomer (PTDA-1000) was used as matrix, the initial transmittance was observed to be relatively higher than that for the lower molecular weight oligomer (PTDA-250). Saturation transmittance for PTDA-1000 was observed at relatively lower voltage than that for PTDA-250, of which transmittance was not saturated even at 60 V. As polymerization temperature increased, the initial transmittance of resulting PDLC film increased due to the larger LC droplets formation and the more matched refractive index between LC and matrix than those cases for the lower polymerization temperature. Though driving voltage decreased for the thinner film, it was considered that optimum thickness of the film should be maintained to get some practical contrast, which is the ratio of off- and on-state transmittance. Furthermore, electro-optic properties such as initial transmittance, driving voltage, and response time were observed to be considerably affected by application of external field during polymerization.

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Improved Genomic DNA Isolation from Soil (토양으로부터 genomic DNA의 효과적인 분리)

  • Kang Ju-Hyung;Kim Bo-Hye;Lee Sun-Yi;Kim Yeong-Jin;Lee Ju-Won;Park Young Min;Ahn Soon-Cheol
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.851-856
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    • 2005
  • Although valuable microbes have been isolated from the soil for the various productions of useful components, the microbes which can be cultivated in the laboratory are only $0.1-1\%$ of all microbes. To solve this problem, the study has recently been tried for making the valuable components from the environment by directly separating unculturable micrbial DNA in the soil. But it is known that humic acid originated from the soil interrupts various restriction enzymes and molecular biological process. Thus, in order to prevent these problems, this study modified the method separated soil DNA with phenol, CTAB and PEG. In order to compare the degree of purity for each DNA and the molecular biological application process, $A_{260}/A_{280}$ ratio, restriction enzymes, and PCR were performed. In case of DNA by the modified method, total yield of DNA was lower but $A_{260}/A_{280}$ ratio was higher than the previously reported methods. It was confirmed that the degree of purity is improved by the modified method. But it was not cut off by all kinds of tested restriction enzymes because of the operation of a very small amount of interrupting substances. When PCR was operated with each diluted DNA in different concentrations and GAPDH primer, the DNA by the modified method could be processed for PCR in the concentration of 100 times higher than by the previously reported separation method. Therefore, this experiment can find out the possibility of utilization for the unknown substances by effectively removing the harmful materials including humic acid and help establishing metagenomic DNA library from the soil DNA having the high degree of purity.

Evaluation of the Diversity of Cyclodextrin-Producing Paenibacillus graminis Strains Isolated from Roots and Rhizospheres of Different Plants by Molecular Methods

  • Vollu Renata Estebanez;Fogel Rafael;Santos Silvia Cristina Cunha dos;Mota Fabio Faria da;Seldin Lucy
    • Journal of Microbiology
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    • v.44 no.6
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    • pp.591-599
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    • 2006
  • To address the diversity of cyclodextrin-producing P. graminis strains isolated from wheat roots and rhizospheres of maize and sorghum sown in Australia, Brazil, and France, restriction fragment length polymorphism analysis of part of genes encoding RNA polymerase (rpoB-RFLP) and DNA gyrase subunit B (gyrB-RFLP) was used to produce genetic fingerprints. A phylogenetic tree based on rpoB gene sequences was also constructed. The isolates originated from Brazil could be separated from those from Australia and France, when data from the rpoB-based phylogenetic tree or gyrB-RFLP were considered. These analyses also allowed the separation of all P. graminis strains studied here into four clusters; one group formed by the strains GJK201 and $RSA19^T$, second group formed by the strains MC22.02 and MC04.21, third group formed by the strains TOD61, TOD 221, TOD302, and TOD111, and forth group formed by all strains isolated from plants sown in Cerrado soil, Brazil. As this last group was formed by strains isolated from sorghum and maize sown in the same soil (Cerrado) in Brazil, our results suggest that the diversity of these P. graminis strains is more affected by the soil type than the plant from where they have been isolated.

A Study on the Properties and Preparation of Silicon-based Defoamer Used in the Purification of Wasted-Water Extruded in the Paper-Fabrication (제지공장의 폐수처리에 사용되는 실리콘계 소포제의 제조 및 물성에 관한 연구)

  • Choi, Sang-goo;Lee, Nae-Taek
    • Applied Chemistry for Engineering
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    • v.16 no.5
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    • pp.614-619
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    • 2005
  • The water-soluble defoamers were fabricated by the mixing polyol, modified silicon resin, silicon resin and surfactant. For the defoamers, the various properties such as phase-separation time, viscosity and defoamerability were examined. The phase-saparation time of PPG mixtures was found to be PPG 400>PPG 3,000>PPG 1000. When PPG 1000 was mixed, mixtures represented the excellent defoamerability. The phase-saparation time of silicon resin mixtures was TSF-451-350>TSF-451-200>TSF-451-50. As more of high molecular silicon resin was mixed, the resulting mixtues showed reduced defoamerability. When the TSF-451-50 was mixed, the mixture's volume was increased because of the reduction of solubility. The modified silicon resin was smoothly dispersed in water, but the compatibility with PPG was not good. The defoamerability of surfactant was SPAN 20>SPAN 60>SPAN 80. SPAN 80 showed good miscibility for the silicon resin, but not good for YAS 6406 or PPG 1000.

Kinetic Studies on Homopolymerization of $\alpha$-Methylstyrene and Sequential Block Copolymerization of Isobutylene with $\alpha$-Methylstyrene by Living/Controlled Cationic Polymerization (리빙/조절 양이온중합에 의한 알파메틸스티렌 호모중합 및 이소부틸렌과의 블록공중합에 대한 반응속도론 연구)

  • Wu, Yibo;Guo, Wenli;Li, Shuxin;Gong, Huiqing
    • Polymer(Korea)
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    • v.32 no.4
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    • pp.366-371
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    • 2008
  • The controlled/living cationic polymerization of $\alpha$-methylstyrene (${\alpha}MeSt$) and sequential block copolymerization of isobutylene (IB) with ${\alpha}MeSt$ were achieved using 2-chloro-2,4,4-trimethylpentane (TMPCl)/titanium tetrachloride ($TiCl_4$)/titanium isopropoxide ($Ti(OiPr)_4$)/2,6-ditert-butylpyridine (DtBP) initiating system in $CH_3Cl$/hexane(50/50 v/v) solvent mixture at $-80^{\circ}C$. The polymerization rate decreased with increasing $[Ti(OiPr)_4]/[TiCl_4]$ ratio in the homopolymerization of ${\alpha}MeSt$. The effects of $[Ti(OiPr)_4]/[TiCl_4]$ ratios and $PIB^+$ molecular weight on the polymerization rate and blocking efficiency were also investigated. Well-defined poly(isobutylene-b-$\alpha$-methylstyrene)s were demonstrated by $^1H$-NMR and triple detection SEC; refractive index (RI), multiangle laser light scattering (MALLS) and ultraviolet (UV) detectors. Blocking efficiencies for the poly(isobutylene-b-$\alpha$-methylstyrene)s of almost 100% were obtained when ${\alpha}MeSt$ was induced by PIB's of $M_n\;{\geq}\;41000$ at $[Ti(OiPr)_4]/[TiCl_4]=1$. Differential scanning calorimetry (DSC) of the block copolymers showed two glass transition temperatures, thereby demonstrating microphase separation.

Cell-SELEX Based Identification of an RNA Aptamer for Escherichia coli and Its Use in Various Detection Formats

  • Dua, Pooja;Ren, Shuo;Lee, Sang Wook;Kim, Joon-Ki;Shin, Hye-su;Jeong, OK-Chan;Kim, Soyoun;Lee, Dong-Ki
    • Molecules and Cells
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    • v.39 no.11
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    • pp.807-813
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    • 2016
  • Escherichia coli are important indicator organisms, used routinely for the monitoring of water and food safety. For quick, sensitive and real-time detection of E. coli we developed a 2'F modified RNA aptamer Ec3, by Cell-SELEX. The 31 nucleotide truncated Ec3 demonstrated improved binding and low nano-molar affinity to E. coli. The aptamer developed by us out-performs the commercial antibody and aptamer used for E. coli detection. Ec3(31) aptamer based E. coli detection was done using three different detection formats and the assay sensitivities were determined. Conventional Ec3(31)-biotin-streptavidin magnetic separation could detect E. coli with a limit of detection of $1.3{\times}10^6CFU/ml$. Although, optical analytic technique, biolayer interferometry, did not improve the sensitivity of detection for whole cells, a very significant improvement in the detection was seen with the E. coli cell lysate ($5{\times}10^4CFU/ml$). Finally we developed Electrochemical Impedance Spectroscopy (EIS) gap capacitance biosensor that has detection limits of $2{\times}10^4CFU/mL$ of E. coli cells, without any labeling and signal amplification techniques. We believe that our developed method can step towards more complex and real sample application.