• Title/Summary/Keyword: Flow Detection

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Detection of Human Taurine Transporter and Production of Monoclonal Antibody

  • An, Hye-Suk;Han, Hee-Chang;Lee, Sun-Min;Park, Taesun;Park, Kun-Koo;Kim, Ha-Won
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2001.11a
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    • pp.102-102
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    • 2001
  • Taurine (2-ethaneaminosulfonic acid) is one of the major intracellular ${\beta}$ -amino acids in mammals and is required for a number of biological processes including membrane stabilization, osmoregulation, antioxidation, detoxification, modulation of calcium flux and neurornodulation. The taurine transporter (TAUT) which contains 12 hydrophobic membrane-spanning domains has been cloned from dog kidney, rat brain, mouse brain, human thyroid, placenta and retina. In this study, The TAUT cDNA from the human intestinal epithelial cell, HT-29 was cloned and sequenced. Reverse-transcription polymerase chain reaction (RT-PCR) was performed to amplify partial cDNA encoding human intestinal TAUT. The coding region of the PCR product was 732 bp long. The primers were designed to encode highly conserved amino acid sequences near the transmembrane domains III (IPYFIFLF) and Ⅵ (KYKYNSYR) both in human and mouse. The TAUT cDNA amplified was ligated into the pGEX 4T-1 expression vector. The resulting sequence of human intestinal TAUT cDNA (Accession number of NCBI Genebank is AF346763) was identical to the sequences of the TAUTs previously determined in the human placenta and retina except 3 base pairs from that of the reported human thyroid. TAUT specific antibodies were generated to use them as biological tools in the studies of the biological role of TAUT. Peptides of 149-162 amino acid residue (14 amino acids) of the TAUT were synthesized. The synthetic peptide used in this study was LFQSFQKELPWAHC. This region was chosen not only to avoid putative glycosylation sites but also to exclude regions of known homology with GABA transporters in the extracellular hydrophilic domains. The synthetic peptide, TAUT-1 was conjugated with carrier protein, kehole lympet hemocyanin (KLH) to use as an antigen. When used for immunization on a rabbit to produce polyclonal antiserum, the conjugates elicited high -titered specific anti-TAUT-1 antibodies, which reacted well with the ovalbumin (OVA) conjugated peptides in ELISA. The KLH-conjugated peptide was also used as immunizing antigen in BALB/c mice to produce TAUT specific monoclonal antibodies. From the culture supernatant of the hybridoma, the specificity of anti-TAUT-1 monoclonal antibodies was confirmed by ELISA. Further applications of more tools in TAUT expression analysis will be performed such as western blotting and flow cytometry.

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Analytical Method Development of Avicularin and Quercitrin in Astilbe chinensis Extract Using HPLC (HPLC를 이용한 노루오줌 추출물의 Avicularin 및 Quercitrin 분석법 개발)

  • Kwon, Jin Gwan;Jung, Yeon Woo;Seo, Changon;Hong, Seong Su;Shin, Hyun Tak;Jung, Su Young;Choi, Jeong June;Kim, Jin Kyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.45 no.4
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    • pp.333-340
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    • 2019
  • This study attempted to eatablish a high performance liquid chromatography (HPLC) analysis method for the determination of avicularin, quercitrin as a part of the quality control for the development of functional cosmetic materials from Astilbe chinensis extract. HPLC was performed on a Capcell Pak C18 MGII column with a gradient elution of 0.05% (v/v) trifluoroacetic acid (TFA) and acetonitrile at a flow rate of 1.0 mL/min at 30 ℃. The analyte was detected at 254 nm. The HPLC method was performed in accordance with the international conference on harmonization (ICH) guideline (version 4, 2005) of analytical procedures with respect to specificity, precision, accuracy, and linearity. The limits of detection and quantitation of avicularin and quercitrin were 0.094 and 0.285 mg/mL, 0.031 and 0.095 mg/mL respectively. Calibration curves showed good linearity r2 > 0.99990 for avicularin and r2 > 0.99994 for quercitrin. Precision of analysis was satisfied with less than 0.59% for avicularin and 0.63% for quercitrin. Recoveries of quantified compounds ranged from 100.97 to 101.77% for avicularin and 100.18 to 100.32% for quercitrin. These result indicated that the established HPLC method is very useful for the determination of marker compounds in A. chinensis extracts.

Development of Method for Analysis of Four Sulfonylurea Pesticides, Rimsulfuron, Ethametsulfuron-methyl, Tribenuron-methyl, Chlorimuron-ethyl Residues by High-Performance Liquid Chromatography with Diode-Array Detection (HPLC/DAD를 이용한 림술푸론, 에타메트설푸론메틸, 트리베누론메칠, 클로리무론에칠 4종 성분의 잔류농약 분석법 개발)

  • Koo, Yun-Chang;Yang, Sung-Yong;Wang, Zeng;An, Eun-Mi;Heo, Kyoung;Kim, Hyeng-Kook;Shin, Han-Seung;Lee, Jin-Won;Lee, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.8
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    • pp.1231-1235
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    • 2010
  • The method for residue analysis of four sulfonylurea pesticides, rimsulfuron, ethametsulfuron-methyl, tribenuron-methyl and chlorimuron-ethyl was examined and analyzed by HPLC with ODS column ($250\;mm{\times}4.6\;mm$, $5\;{\mu}m$ diameter particle size) which was maintained at $35^{\circ}C$. Mobile phase consisted of solvent A (20 mM $KH_2PO_4$, pH 2.5) and solvent B (acetonitrile). Isocratic elution of the column with 45% solvent A and 55% solvent B at a flow rate of 1 mL/min resulted in retention times of 5.92, 6.54, 9.28, and 14.35 min for rimsulfuron, ethametsulfuron-methyl, tribenuron-methyl, and chlorimuron-ethyl, respectively. All injection volumes were $20\;{\mu}L$. The limit of quantitation was 0.02, 0.01, 0.001, and 0.004 mg/kg for rimsulfuron, ethametsulfuron-methyl, tribenuron-methyl, and chlorimuron-ethyl, respectively. Recovery rate test was performed with three farm products, rice, apple and soybean. Four sulfonylurea pesticides were spiked at concentrations of 0.05, 0.1 and 0.5 mg/kg. The recovery rates were ranged from 86.12% to 116.26% and the standard deviations of all experiments were within 10%.

Clinical Applications of Neuroimaging with Susceptibility Weighted Imaging: Review Article (SWI의 신경영상분야의 임상적 이용)

  • Roh, Keuntak;Kang, Hyunkoo;Kim, Injoong
    • Investigative Magnetic Resonance Imaging
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    • v.18 no.4
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    • pp.290-302
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    • 2014
  • Purpose : Susceptibility-weighted magnetic resonance (MR) sequence is three-dimensional (3D), spoiled gradient-echo pulse sequences that provide a high sensitivity for the detection of blood degradation products, calcifications, and iron deposits. This pictorial review is aimed at illustrating and discussing its main clinical applications. Materials and Methods: SWI is based on high-resolution, 3D, fully velocity-compensated gradient-echo sequences using both magnitude and phase images. To enhance the visibility of the venous structures, the magnitude images are multiplied with a phase mask generated from the filtered phase data, which are displayed at best after post-processing of the 3D dataset with the minimal intensity projection algorithm. A total of 200 patients underwent MR examinations that included SWI on a 3 tesla MR imager were enrolled. Results: SWI is very useful in detecting multiple brain disorders. Among the 200 patients, 80 showed developmental venous anomaly, 22 showed cavernous malformation, 12 showed calcifications in various conditions, 21 showed cerebrovascular accident with susceptibility vessel sign or microbleeds, 52 showed brain tumors, 2 showed diffuse axonal injury, 3 showed arteriovenous malformation, 5 showed dural arteriovenous fistula, 1 showed moyamoya disease, and 2 showed Parkinson's disease. Conclusion: SWI is useful in detecting occult low flow vascular lesions, calcification and microbleed and characterising diverse brain disorders.

Highly Sensitive Detection of Pathogenic Bacteria Using PDMS Micro Chip Containing Glass Bead (유리비드를 포함한 PDMS 마이크로칩을 이용한 고감도 감염성 병원균 측정에 관한 연구)

  • Won, Ji-Yeong;Min, Jun-Hong
    • KSBB Journal
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    • v.24 no.5
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    • pp.432-438
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    • 2009
  • Here, we demonstrated simple nucleic acid, RNA, concentration method using polymer micro chip containing glass bead ($100\;{\mu}m$). Polymer micro chip was fabricated by PDMS ($1.5\;cm\;{\times}\;1.5\;cm$, $100\;{\mu}m$ in the height) including pillar structure ($160\;{\mu}m\;(I)\;{\times}\;80\;{\mu}m\;(w)\;{\times}\;100\;{\mu}m\;(h)$, gap size $50\;{\mu}m$) for blocking micro bead. RNA could be adsorbed on micro glass bead at low pH by hydrogen bonding whereas RNA was released at high pH by electrostatic force between silica surface and RNA. Amount of glass beads and flow rate were optimized in aspects of adsorption and desorption of RNA. Adsorption and desorption rate was measured with real time PCR. This concentrated RNA was applied to amplification micro chip in which NASBA (Nucleic Acid Sequence Based Amplification) was performed. As a result, E.coli O157 : H7 in the concentration of 10 c.f.u./10 mL was successfully detected by these serial processes (concentration and amplification) with polymer micro chips. It implies this simple concentration method using polymer micro chip can be directly applied to ultra sensitive method to measure viable bacteria and virus in clinical samples as well as environmental samples.

An Analysis Method for Detecting Vulnerability to Symbolic Link Exploit (심볼릭 링크 공격 취약성 검출을 위한 분석 기법)

  • Joo, Seong-Yong;Ahn, Joon-Seon;Jo, Jang-Wu
    • The KIPS Transactions:PartA
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    • v.15A no.1
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    • pp.45-52
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    • 2008
  • In this paper we define a vulnerable code to symbolic link exploit and propose a technique to detect this using program analysis. The existing methods to solve symbolic link exploit is for protecting it, on accessing a temporary file they should perform an investigation whether the file is attacked by symbolic link exploit. If programmers miss the investigation, the program may be revealed to symbolic link exploit. Because our technique detects all the vulnerable codes to symbolic link exploit, it helps programmers keep the program safety. Our technique add two type qualifiers to the existing type system to analyze vulnerable codes to symbolic link exploit, it detects the vulnerable codes using type checking including the added type qualifiers. Our technique detects all the vulnerable codes to symbolic link exploit automatically, it has the advantage of saving costs of modifying and of overviewing all codes because programmers apply the methods protecting symbolic link exploit to only the detected codes as vulnerable. We experiment our analyzer with widely used programs. In our experiments only a portion of all the function fopen() is analyzed as the vulnerabilities to symbolic link exploit. It shows that our technique is useful to diminish modifying codes.

Determination of Rare Earth Elements by Inductively Coupled Plasma Mass Spectrometry with Introduction of Organic Phase Using Ultrasonic Nebulizer (초음파 분무기를 이용한 유기상의 주입에 의한 희토류 원소의 유도결합 플라스마 질량분석법적 정량)

  • Lee, Sang Hak;Son, Bum Mok
    • Analytical Science and Technology
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    • v.15 no.2
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    • pp.127-134
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    • 2002
  • Analytical method to determine rare earth elements which were extracted to organic phase by inductively coupled plasma mass spectrometry(ICP-MS) was investigated. Organic phase which had extracted rare earth elements was directly aspirated into ICP-MS by ultrasonic nebulizer(USN) in order to reduce solvent load to the plasma. Then, the count rate increased when MIBK(methyl isobutyl ketone) was added to EtOH(ethanol) but decreased when 0.03 M HEH(2-ethylhexyl-2-ethylhexyl phosphonic acid, $P_{507}$, PC88A) was added to solvent which mixed MIBK with EtOH. The optimal temperatures of desolvation system were -10 $^{\circ}C$ for the condenser and 150 $^{\circ}C$ for the heating tube. The optimal nebulizer flow rate which gave maximum count rate and minimum reflect power was 0.7 L/min. The optimal pH and extraction time were 4.3 and 10 min for MIBK-0.03 M HEH system. Detection limits which were obtained through calibration curves at the range of 0.2 ${\sim}$ 20 ng/mL were 0.02 ${\sim}$ 0.05 ng/mL under the optimal experimental conditions.

Determination and preconcentration of Cu(II) using microcrystalline p-Dichlorobenzene loaded with salicylaldoxime (Salicylaldoxime이 내포된 p-Dichlorobenzene 미세결정을 이용한 Cu(II)의 예비농축 및 정량)

  • Lee, Ha-Na;Choi, Hee-Seon
    • Analytical Science and Technology
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    • v.23 no.3
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    • pp.240-246
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    • 2010
  • A technique for the determination of trace Cu(II) in various real samples by FAAS after the column preconcentration onto p-dichlorobenzene-SA adsorbent, which is microcrystalline p-dichlorobenzene loaded with salicylaldoxime (SA) has been developed. Several experimental conditions such as pH of the sample solution, the amount of chelating agent salicylaldoxime, the amount of adsorbent p-dichlorobenzene-SA, and flow rate of sample solution were optimized. The interfering effects of various concomitant ions were investigated. $CN^-$ interfered more seriously than any other ions. However, the interference by $1\;{\mu}g\;mL^{-1}\;CN^-$ could be overcome completely by controlling the concentration of Ni(II) to $20\;{\mu}g\;mL^{-1}$. The linear range, correlation coefficient ($R^2$) and detection limit obtained by this technique were $3.0\sim100\;ng\;mL^{-1}$, 0.9901, and $3.1\;ng\;mL^{-1}$, respectively. For validating this technique, the aqueous samples (wastewater, reservoir water and stream water) and the food samples (orange juice, fresh egg and skim milk) were used. Recovery yields of 93~104% were obtained. These measured mean values were not differents from ICP-MS data at 95% confidence level. The good results were obtained from the experiments using the rice flour certified reference material (CRM) sample. Based on the experimental results, it was found that this technique could be applied to the preconcentration and determination of Cu(II) for various real samples.

Establishment of content criteria of marker compounds through the monitoring of Achyranthis Radix collected from Korea and China (한국 및 중국 지역에서 수집된 우슬의 모니터링을 통한 지표성분의 함량기준 설정)

  • Kim, Dae-Hyun;Kim, Sang-Hyuk;Jang, Yu-Seon;Shin, Min-Chul;Chu, Van Men;Lee, Young-Keun;Woo, Mi-Hee;Kang, Jong-Seong
    • Analytical Science and Technology
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    • v.25 no.4
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    • pp.250-256
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    • 2012
  • Two marker compounds of Achyranthis Radix, ecdysterone and inokosterone, were analyzed by HPLC on an ODS column ($250{\times}4.6$ mm, 5 ${\mu}m$) with a mobile phase of 15% acetonitrile containing 0.08% formic acid at a flow rate of 1.0 mL/min and a detection wavelength of UV 254 nm. The method was validated by ICH guideline and applied to the monitoring of marker compounds in 93 samples of Achyranthis Radix collected at various areas in Korea and China. The new content criteria of ecdysterone and inokosterone, established using linear regression method were 0.033% and 0.020%, respectively. When the new content criteria were applied to the quality control test of commercial Achyranthis Radix, 95.4% of total samples including 100% of Korean and 92.6% of Chinese samples were passed the test. Application of new content criteria could protect the Korean products and decrease the distribution of Chinese products with lower quality.

The Change of Tartrate Resistant Acid Phosphatase Activity in Capsaicin-Induced Canine Chondrocyte Death (개 연골세포의 손상에 의한 Tartrate Resistant Acid Phosphatase 활성의 변화 측정)

  • Seol Jae-Won;Lee Hae-Beom;Kim Nam-Soo;Kim In-Shik;Kang Hyung-Sub;Lee Young-Hoon;Kang Dong-Won;Park Sang-Youel
    • Journal of Veterinary Clinics
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    • v.23 no.2
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    • pp.144-148
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    • 2006
  • Apoptotic death of articular chondrocytes has been implicated in the pathogenesis of osteoarthritis. Tartrate resistant acid phosphatase (TRAP) has been used for several years as a marker enzyme of bone-resorbing osteoclasts. This study investigated the activity of TRAP in media of apoptotic cell death-induced canine chondrocyte. We exposed canine chondrocyte to capsaicin and the results showed that capsaicin induced cell death in a dose dependent manner. And we measured TRAP activity in media of chondrocyte death induced by capsaicin treatment and the results capsaicin significantly increased the activity of TRAP in media for dose dependent. We also investigated whether the combination treatment with capsaicin and TRAIL enhance apoptotic cell death in canine chondrocyte. We exposed canine chondrocyte to capsaicin for 24 hrs at the indicated dose, and then treated with recombinant TRAIL protein for 24 hrs. TRAIL alone did not induce cell death after 24 hours, but the combined treatment of both induced more cell death compared with capsaicin alone in a dose dependent manner. Also, the combination treatment with capsaicin and TRAIL increased the activity of TRAP in culture media. These results suggest that TRAP can flow out into extracellular after chondrocyte damage, and TRAP may be a successful biomarker for detection of joint disease such as osteoarthritis.