• Title/Summary/Keyword: Reaction monitoring

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Quantitative measurement of salivary testosterone in Korean adults by stable isotope-dilution liquid chromatographyelectrospray-tandem mass spectrometry

  • Lee, Sang-Hoo;Kwon, Soon-Ho;Shin, Hye-Jin;Park, Ji-Myeong;Lim, Hwan-Sub;Lee, Kyoung-Ryul;Kim, Young-Jin
    • BMB Reports
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    • v.43 no.11
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    • pp.761-765
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    • 2010
  • Salivary testosterone levels in Korean adults were quantitatively measured for the first time by liquid chromatography-electrospray-tandem mass spectrometry (LC ESI MS/MS). Salivary testosterone was separated on a multiple reaction monitoring (MRM) chromatogram within 7 min. The LC ESI MS/MS assay was validated over the linearity range of 0.01-2.00 ng/ml (r=0.99987) using testosterone-$d_3$ as an internal standard. The lower limit of quantification (LOQ) was 0.01 ng/ml. The intra- and inter-assay precisions were 1.54% to 4.09% and 0.96% to 4.29%, respectively. The mean recovery was 93.32% (range 88.43-98.05%). The validated assay was then applied to measure the salivary testosterone levels of Korean adults. In men, the salivary testosterone level collected between 9:00-11:00 am was approximately 2.8 times higher than that in women (P < 0.0001). Salivary testosterone levels in both sexes negatively correlated with age. The present assay would also be useful in measuring salivary testosterone levels in clinical laboratories.

THE EFFECTS OF CELL WALL PROTEINS OF STREPTOCOCCUS SPP. ON DNA SYNTHESIS OF L929 CELLS AND THEIR SDS-PAGE PATTERNS (연쇄 구균의 세포벽 단백질이 L929 세포의 DNA합성에 미치는 영향 및 SDS-PAGE 양상에 관한 연구)

  • Lee, Se-Jong;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
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    • v.20 no.1
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    • pp.71-95
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    • 1995
  • Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

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Monitoring of Genetically Modified Bean Sprouts at Traditional Markets in Seoul and a Survey of Perception of Traditional Market Merchants and High School Students on Genetically Modified Organisms (서울 일부지역 재래시장의 유전자재조합 콩나물의 판매실태 및 상인과 일부 고등학생의 유전자재조합 식품에 대한 인식도 조사)

  • Choi, Seul-Ki;Lim, Jung-Hoon;Shin, Jee-Seong;Jeong, Jin-An;Joung, Hyo-Jee
    • Korean Journal of Community Nutrition
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    • v.15 no.5
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    • pp.679-686
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    • 2010
  • The purpose of this study is to monitor genetically modified bean sprouts at traditional markets in Seoul and to investigate perception of traditional market merchants and high school students on genetically modified organisms (GMOs). We analyzed 30 bean sprouts that were selected at 11 traditional markets in Seoul using the method of polymerase chain reaction (PCR). Also, we compared perception of GMOs between merchants (n = 30) and students (n = 126). Knowledge test about GMOs was performed by students. The result of PCR, 16.7% of bean sprouts were confirmed as GM bean sprouts (n = 5). Students had significantly more exposure to information about GMOs than merchants (p < 0.05). Major information sources about GMOs were from mass media (television, newspaper and radio). About half of subjects were not aware that they eat GMOs and GMOs are sold to consumers. Only 17.3% of subjects had constant eating intent for GMOs after perceiving foods that he/she usually eats are GMOs. 51.3% of subjects had willingness to purchase GMOs if GMOs have same quality and lower cost than natural foods. 37.2% of subjects thought that GMOs would be harmful to humans. Students had more positive perception of GMOs' side effects than merchants (p < 0.01). There was no merchant who knew labeling of GMOs. 19.1% of students knew labeling of GMOs. Students' mean percentage of correct answers of six questions about GMOs was 45.2%. Therefore, providing adequate information about GMOs is needed for consumer's choice whether to purchase GMOs or not.

Quantitative Determination of the Bioactive Marker Components in Gyeji-tang Using LC-ESI-MS/MS (LC-ESI-MS/MS를 이용한 계지탕 중 주요 성분 분석)

  • Seo, Chang-Seob;Ha, Hyekyung
    • Korean Journal of Pharmacognosy
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    • v.49 no.1
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    • pp.76-83
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    • 2018
  • A traditional herbal formula, Gyeji-tang has been used to treat the early colds, headache, chills, and fever in Asian countries. In this study, we were performed simultaneous determination of the 14 bioactive marker compounds, gallic acid, spinosin, paeoniflorin, albiflorin, liquiritin apioside, liquiritin, 6'''-feruloylspinosin, liquilitigenin, coumarin, cinnmamic acid, benzoylpaeoniflorin, cinnamaldehyde, glycyrrhizin, and 6-gingerol in Gyeji-tang using an ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS/MS). Analytical column was used a Waters Acquity UPLC BEH $C_{18}$ analytical column ($2.1{\times}100mm$, $1.7{\mu}m$) and maintained at $45^{\circ}C$ with a flow rate of 0.3 mL/min. The mobile phase consists of 0.1% (v/v) formic acid in water and acetonitrile with gradient elution. The MS analysis was conducted using multiple reaction monitoring in the positive and negative modes by a Waters ACQUITY TQD LC-MS/MS system. The calibration curves of 14 bioactive marker compounds showed linearity with correlation coefficients ${\geq}0.9798$. The limits of detection and quantification values were in the range of 0.11-6.66 ng/mL and 0.34-19.99 ng/mL, respectively. As a result of the analysis using the established LC-MS/MS method, the amounts of tested 14 compounds in the lyophilized Gyeji-tang sample were detected up to $85.7{\mu}g/g$. These results may be useful for quality assessment of a traditional herbal formulas.

HPLC-tandem Mass Spectrometric Analysis of the Marker Compounds in Forsythiae Fructus and Multivariate Analysis

  • Cho, Hwang-Eui;Ahn, Su-Youn;Son, In-Seop;Hwang, Gyung-Hwa;Kim, Sun-Chun;Woo, Mi-Hee;Lee, Seung-Ho;Son, Jong-Keun;Hong, Jin-Tae;Moon, Dong-Cheul
    • Natural Product Sciences
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    • v.17 no.2
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    • pp.147-159
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    • 2011
  • A high-performance liquid chromatography-electrospray ionization-tandem mass spectrometric method was developed to determine simultaneously eight marker constituents of Forsythiae fructus, and subsequently applied it to classify its two botanical origins. The marker compounds of Forsythia suspensa were phillyrin, pinoresinol, phillygenin, lariciresinol and forsythiaside; those of F.viridissima were arctiin, arctigenin and matairesinol. Separation of the eight analytes was achieved on a phenyl-hexyl column (150${\times}$2.0 mm i.d., 3 ${\mu}M$) using gradient elution with the mobile phase: (A) 10% acetonitrile in 0.5% acetic acid, (B) 40% aqueous acetonitrile. A few fragment ions specific to the types of lignans, among the product ions generated by collisonally induced dissociation (CID) of molecular ion clusters, such as [M-H]$^-$ or [M+OAc]$^-$ were used not only for fingerprinting analysis but for the quantification of each epimer by using multiple-reaction monitoring mode. It was shown good linearity ($r^2{\geq}$ 0.9998) over the wide range of all analytes; intra- and inter-day precisions (RSD, %) were within 9.14% and the accuracy ranged from 84.3 to 115.1%. The analytical results of 40 drug samples, combined with multivariate statistical analyses - principal component analysis (PCA) and hierarchical cluster analysis (HCA) - clearly demonstrated the classification of the test samples according to their botanical origins. This method would provide a practical strategy for assessing the authenticity or quality of the herbal drug.

HPLC/MS/MS Method for Determination of Soyasaponins in the Soybean Varieties (콩 함유 사포닌의 종류 및 함량 분석)

  • Han, Sang-Jun
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.56 no.3
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    • pp.244-249
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    • 2011
  • A sensitive and rapid high-performance liquid chromatographic-tandem mass spectrometric (HPLC/MS/MS) assay was developed for the determination of soyasaponins in soybean. Among soyasaponins, soyasaponin I was isolated and characterized from methanol extracts of soybean as analytical standards and the development of a new analytical procedure for quantification of its content in various cultivars. The structures of these compound was elucidated by $^1H$, $^{13}C$ NMR experiments and by mass spectrometric analysis. Aqueous ethanol extracts of soybean samples were injected on an Agilent XDB-C18 column ($4.6mm{\times}50mm$, $1.8{\mu}m$) with a mobile phase consisting of 10 mM ammonium acetate-acetonitrile, a flow rate of 0.3 mL/min and a total run time of 8 min. Detection was performed by mass spectrometer bin the multiple reaction monitoring (MRM) mode with negative electrospray ionization (ESI) m/z at 941 ${\rightarrow}$ 615 for soyasaponin I. In the 9 soybean samples, contents of soyasaponin I ranged from 205 to 726 mg/kg, and correlated negatively with seed size.

Analysis of X Irradiation Related Genes in HL60 Cells Using cDNA Microarray (cDNA Microarray를 이용한 HL60 세포주에서 방사선 조사 관련 유전자의 검색 및 분석)

  • Park, Keon-Uk;Hwang, Mi-Sun;Suh, Seong-Il;Suh, Min-Ho;Kwon, Taeg-Kyu;Park, Jong-Wook;Cho, Jae-We;Choi, Eun-Ju;Baek, Won-Ki
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.4
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    • pp.299-308
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    • 2000
  • Recently developed cDNA microarray or DNA chip technology allows expression monitoring of expression of hundreds and thousands of genes simultaneously and provides a format for identifying genes as well as changes in their activity. In order to search for changes in gene expression after X irradiation in HL60 cells, cDNA microarray technique was done. In this study, expression of 588 human genes (including oncogenes, tumor suppressor genes, cell cycle regulator genes, intracellular signal transduction modulator genes, apoptosis related genes, transcription factor genes, growth factors and receptor genes, cytokine genes, etc) were analyzed. For cDNA microarray analysis mRNAs were extracted from control and 8 Gy-irradiated HL60 cells. As a result the changes in expression of several genes were observed. This alteration of gene expression was confirmed by reverse transcription-polymerase chain reaction. The expression of heat shock 60 KD protein, c-jun, erythroid differentiation factor, CPP32, myeloid cell nuclear differentiation antigen, MAP kinase-activated protein kinase, interleukin-8, monocyte chemotactic peptide 1 and RANTES genes was increased, but the expression of p55CDC gene was decreased after X irradiation.

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MWCNT, silver nanoparticles, CuBTC를 사용한 염소 이온 센서 합성

  • Gwak, Byeong-Gwan;Park, Su-Bin;Yu, Bong-Yeong
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2018.06a
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    • pp.101-101
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    • 2018
  • Quantitative measurement of chloride ion concentration has an important role in various fields of electrochemistry, medical science, biology, metallurgy, architecture, etc. Among them, its importance of architecture is ever-growing due to unexpected degradations of building structure. These situations are caused by corrosion of reinforced concrete (RC) structure of buildings. And chloride ions are the most powerful factors of RC structure corrosion. Therefore, precise inspection of chloride ion concentration must be required to increase the accuracy of durability monitoring. Multi-walled Carbon nanotubes (MWCNTs) have high chemical resistivity, large surface area and superior electrical property. Thus, it is suitable for the channels of electrical signals made by the sensor. Silver nanoparticles were added to giving the sensing property. CuBTC, one of the metal organic frameworks (MOFs), was employed as a material to improve the sensing property because of its hydrophilicity and high surface area to volume ratio. In this study, sensing element was synthesized by various chemical reaction procedures. At first, MWCNTs were functionalized with a mixture of sulfuric acid and nitric acid because of enhancement of solubility in solution and surface activation. And functionalized MWCNTs, silver nanoparticles, and CuBTC were synthesized on PTFE membrane, one by one. Electroless deposition process was performed to deposit the silver nanoparticles. CuBTC was produced by room temperature synthesis. Surface morphology and composition analysis were characterized by scanning electron microscope (SEM), energy dispersive X-ray spectroscopy (EDS), respectively. X-ray photoelectron spectroscopy (XPS) was also performed to confirm the existence of sensing materials. The electrical properties of sensor were measured by semiconductor analyzer. The chloride ion sensing characteristics were confirmed with the variation of the resistance at 1 V.

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Perioperative red blood cell transfusion in orofacial surgery

  • Park, So-Young;Seo, Kwang-Suk;Karm, Myong-Hwan
    • Journal of Dental Anesthesia and Pain Medicine
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    • v.17 no.3
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    • pp.163-181
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    • 2017
  • In the field of orofacial surgery, a red blood cell transfusion (RBCT) is occasionally required during double jaw and oral cancer surgery. However, the question remains whether the effect of RBCT during the perioperative period is beneficial or harmful. The answer to this question remains challenging. In the field of orofacial surgery, transfusion is performed for the purpose of oxygen transfer to hypoxic tissues and plasma volume expansion when there is bleeding. However, there are various risks, such as infectious complications (viral and bacterial), transfusion-related acute lung injury, ABO and non-ABO associated hemolytic transfusion reactions, febrile non-hemolytic transfusion reactions, transfusion associated graft-versus-host disease, transfusion associated circulatory overload, and hypersensitivity transfusion reaction including anaphylaxis and transfusion-related immune-modulation. Many studies and guidelines have suggested RBCT is considered when hemoglobin levels recorded are 7 g/dL for general patients and 8-9 g/dL for patients with cardiovascular disease or hemodynamically unstable patients. However, RBCT is occasionally an essential treatment during surgeries and it is often required in emergency cases. We need to comprehensively consider postoperative bleeding, different clinical situations, the level of intra- and postoperative patient monitoring, and various problems that may arise from a transfusion, in the perspective of patient safety. Since orofacial surgery has an especially high risk of bleeding due to the complex structures involved and the extensive vascular distribution, measures to prevent bleeding should be taken and the conditions for a transfusion should be optimized and appropriate in order to promote patient safety.

The Applicability Evaluation of UV Indicators in the Exhibition Environment (전시환경에 대한 자외선 지시카드 적용성 평가)

  • Lim, BoA;Shin, EunJeong;Kim, MyoungNam;Lee, SunMyung
    • 보존과학연구
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    • s.33
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    • pp.85-97
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    • 2012
  • In this study, reliability assessment was carried out on UV(ultraviolet) indicators for the application of the indicator method on cultural heritages. As a result, some UV indicators(L, K, and M) proved to be unsuitable due to their low reactivity to UV and high reactivity to RH(relative humidity). On the other hand, indicator S showed considerable color changes and fading due to UV-B and UV-C according to the integral amount of UV irradiation. Indicator S showed the most distinct reaction to UV and a remarkable applicability to exhibition light, which are used in museums. Therefore, UV indicator S is an easy and useful monitoring tool for light environment management in cultural- property conservation facilities.

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