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Full validation of high-throughput bioanalytical method for the new drug in plasma by LC-MS/MS and its applicability to toxicokinetic analysis

  • Han, Sang-Beom
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2006.11a
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    • pp.65-74
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    • 2006
  • Modem drug discovery requires rapid pharmacokinetic evaluation of chemically diverse compounds for early candidate selection. This demands the development of analytical methods that offer high-throughput of samples. Naturally, liquid chromatography / tandem mass spectrometry (LC-MS/MS) is choice of the analytical method because of its superior sensitivity and selectivity. As a result of the short analysis time(typically 3-5min) by LC-MS/MS, sample preparation has become the rate- determining step in the whole analytical cycle. Consequently tremendous efforts are being made to speed up and automate this step. In a typical automated 96-well SPE(solid-phase extraction) procedure, plasma samples are transferred to the 96-well SPE plate, internal standard and aqueous buffer solutions are added and then vacuum is applied using the robotic liquid handling system. It takes only 20-90 min to process 96 samples by automated SPE and the analyst is physically occupied for only approximately 10 min. Recently, the ultra-high flow rate liquid chromatography (turbulent-flow chromatography)has sparked a huge interest for rapid and direct quantitation of drugs in plasma. There is no sample preparation except for sample aliquotting, internal standard addition and centrifugation. This type of analysis is achieved by using a small diameter column with a large particle size(30-5O ${\mu}$m) and a high flow rate, typically between 3-5 ml/min. Silica-based monolithic HPLC columns contain a novel chromatographic support in which the traditional particulate packing has been replaced with a single, continuous network (monolith) of pcrous silica. The main advantage of such a network is decreased backpressure due to macropores (2 ${\mu}$m) throughout the network. This allows high flow rates, and hence fast analyses that are unattainable with traditional particulate columns. The reduction of particle diameter in HPLC results in increased column efficiency. use of small particles (<2 urn), however, requires p.essu.es beyond the traditional 6,000 psi of conventional pumping devices. Instrumental development in recent years has resulted in pumping devices capable of handling the requirements of columns packed with small particles. The staggered parallel HPLC system consists of four fully independent binary HPLC pumps, a modified auto sampler, and a series of switching and selector valves all controlled by a single computer program. The system improves sample throughput without sacrificing chromatographic separation or data quality. Sample throughput can be increased nearly four-fold without requiring significant changes in current analytical procedures. The process of Bioanalytical Method Validation is required by the FDA to assess and verify the performance of a chronlatographic method prior to its application in sample analysis. The validation should address the selectivity, linearity, accuracy, precision and stability of the method. This presentation will provide all overview of the work required to accomplish a full validation and show how a chromatographic method is suitable for toxirokinetic sample analysis. A liquid chromatography/tandem mass spectrometry (LC-MS/MS) method developed to quantitate drug levels in dog plasma will be used as an example of tile process.

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Characterization and Formation Mechanism of Zr-Cu and Zr-Cu-Al Metallic Glass Thin Film by Sputtering Process

  • Lee, Chang-Hun;Sun, Ju-Hyun;Moon, Kyoung-Il;Shin, Seung-Yong
    • Proceedings of the Korean Vacuum Society Conference
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    • 2012.02a
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    • pp.271-272
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    • 2012
  • Bulk Metallic Glasses (BMGs or amorphous alloy) exhibit high strength and good corrosion resistance. Applications of thin films and micro parts of BMGs have been used a lot since its inception in the research of BMGs. However, Application and fabrication of BMGs are limited to make structural materials. Thin films of BMGs which is sputtered on the surface of structural materials by sputtering process is used to improve limits about application of BMGs. In order to investigate the difference of properties between designed alloys and thin films, we identified that thin films deposited on the surface that have the characteristic of the amorphous films and the composition of designed alloys. Zr-Cu (Cu=30, 35, 38, 40, 50 at.%) and Zr-Cu-Al (Al=10 at.% fixed, Cu=26, 30, 34, 38 at.%) alloys were fabricated with Zr (99.7% purity), Cu (99.997% purity), and Al (99.99% purity) as melting 5 times by arc melting method before rods 2mm in diameter was manufactured. In order to analyze GFA (Glass Forming Ability), rods were observed by Optical Microscopy and SEM and $T_g$, $T_x$, ($T_x$ is crystallization temperature and $T_g$ is the glass transition temperature) and Tm were measured by DTA and DSC. Powder was manufactured by Gas Atomizer and target was sintered using powder in large supercooled liquid region ($=T_x-T_g$) by SPS(Spark Plasma Sintering). Amorphous foil was prepared by RSP process with 5 gram alloy button. The composition of the foil and sputtered thin film was analyzed by EDS and EPMA. In the result of DSC curve, binary alloys ($Zr_{62}Cu_{38}$, $Zr_{60}Cu_{40}$, $Zr_{50}Cu_{50}$) and ternary alloys ($Zr_{64}Al_{10}Cu_{26}$, $Zr_{56}Al_{10}Cu_{34}$, $Zr_{52}Al_{10}Cu_{38}$) have $T_g$ except for $Zr_{70}Cu_{30}$ and $Zr_{60}Al_{10}Cu_{30}$. The compositions with $T_g$ made into powders. Figure shows XRD data of thin film showed similar hollow peak.

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Text Filtering using Iterative Boosting Algorithms (반복적 부스팅 학습을 이용한 문서 여과)

  • Hahn, Sang-Youn;Zang, Byoung-Tak
    • Journal of KIISE:Software and Applications
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    • v.29 no.4
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    • pp.270-277
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    • 2002
  • Text filtering is a task of deciding whether a document has relevance to a specified topic. As Internet and Web becomes wide-spread and the number of documents delivered by e-mail explosively grows the importance of text filtering increases as well. The aim of this paper is to improve the accuracy of text filtering systems by using machine learning techniques. We apply AdaBoost algorithms to the filtering task. An AdaBoost algorithm generates and combines a series of simple hypotheses. Each of the hypotheses decides the relevance of a document to a topic on the basis of whether or not the document includes a certain word. We begin with an existing AdaBoost algorithm which uses weak hypotheses with their output of 1 or -1. Then we extend the algorithm to use weak hypotheses with real-valued outputs which was proposed recently to improve error reduction rates and final filtering performance. Next, we attempt to achieve further improvement in the AdaBoost's performance by first setting weights randomly according to the continuous Poisson distribution, executing AdaBoost, repeating these steps several times, and then combining all the hypotheses learned. This has the effect of mitigating the ovefitting problem which may occur when learning from a small number of data. Experiments have been performed on the real document collections used in TREC-8, a well-established text retrieval contest. This dataset includes Financial Times articles from 1992 to 1994. The experimental results show that AdaBoost with real-valued hypotheses outperforms AdaBoost with binary-valued hypotheses, and that AdaBoost iterated with random weights further improves filtering accuracy. Comparison results of all the participants of the TREC-8 filtering task are also provided.

Intermediate-Representation Translation Techniques to Improve Vulnerability Analysis Efficiency for Binary Files in Embedded Devices (임베디드 기기 바이너리 취약점 분석 효율성 제고를 위한 중간어 변환 기술)

  • Jeoung, Byeoung Ho;Kim, Yong Hyuk;Bae, Sung il;Im, Eul Gyu
    • Smart Media Journal
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    • v.7 no.1
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    • pp.37-44
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    • 2018
  • Utilizing sequence control and numerical computing, embedded devices are used in a variety of automated systems, including those at industrial sites, in accordance with their control program. Since embedded devices are used as a control system in corporate industrial complexes, nuclear power plants and public transport infrastructure nowadays, deliberate attacks on them can cause significant economic and social damages. Most attacks aimed at embedded devices are data-coded, code-modulated, and control-programmed. The control programs for industry-automated embedded devices are designed to represent circuit structures, unlike common programming languages, and most industrial automation control programs are designed with a graphical language, LAD, which is difficult to process static analysis. Because of these characteristics, the vulnerability analysis and security related studies for industry automation control programs have only progressed up to the formal verification, real-time monitoring levels. Furthermore, the static analysis of industrial automation control programs, which can detect vulnerabilities in advance and prepare for attacks, stays poorly researched. Therefore, this study suggests a method to present a discussion on an industry automation control program designed to represent the circuit structure to increase the efficiency of static analysis of embedded industrial automation programs. It also proposes a medium term translation technology exploiting LLVM IR to comprehensively analyze the industrial automation control programs of various manufacturers. By using LLVM IR, it is possible to perform integrated analysis on dynamic analysis. In this study, a prototype program that converts to a logical expression type of medium language was developed with regards to the S company's control program in order to verify our method.

A Experimental Study of PTEN (Phosphatase and Tensin) Role in Mesothelioma (중피종에서 PTEN(Phosphatase and Tensin)의 역할에 대한 실험적 연구)

  • 이석기;김권천
    • Journal of Chest Surgery
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    • v.36 no.11
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    • pp.852-857
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    • 2003
  • Background: Conventional treatment for mesothelioma is largely ineffective. We evaluated the novel approach of adenoviral gene transfection of PTEN gene in mesothelioma cancer cell lines, inflammatory and epithelial subtype, which are sensitive to adenoviral p53. Material and Method: Binary adenoviral PTEN and LacZ (Ad/GT-LacZ and Ad/GV16) vectors were used for transduction of the mesothelioma cell lines, REN (p53 sensitive). Protein levels were determined by Western blotting assay. Apoptosis was assessed by fluorescence-activated cell sorter analysis of subdiploid populations. Cell viability was determined with the XTT assay. Statistical analysis was performed with analysis of variance and the Student t test. Result: 72 hours after the treatment of adenoviral PTEN gene, cell killing were 32.9% for REN compared to control cell (2.5%) at MOI of 20. Also we observed the over-expression of proapoptotic protein, bax and decreased expression of bcl-2 protein in REN cells. But the expression of BCL-xl, Bak, Bad proteins were not altered. Conclusion: Adenovirus Pten-mediated overexpression of the Bax gene induces apoptosis and decreased cellular viability in p53-sensitive mesothelioma cells. These data suggest that the transfection of PTEN gene may represent a alternative gene therapy strategy to treat mesothelioma.

Excess Volumes of Aqueous Solutions of Nonionic Amphiphile 2-(2-Hexyloxyethoxy)ethanol (비이온 양친매성 분자 2-(2-Hexyloxyethoxy)ethanol 수용액의 과잉 부피)

  • Lee, Jungno;Hwang, Yoonmi;Kang, Minhee;Lim, Kyung-Hee
    • Journal of the Korean Applied Science and Technology
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    • v.37 no.3
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    • pp.516-525
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    • 2020
  • The densities of aqueous solutions of the amphiphile 2-(2-hexyloxyethoxy)ethanol (C6E2) were measured at 279.15 K and 282.15 K by vibrating-tube densitometry. Then using the density data of the binary C6E2 (1)/water(2) system, the excess volumes and partial molar volumes were determined at various compositions. Excess volume VE exhibits negative deviation for the whole region of composition, which implies relatively stronger attraction between molecules. At the C6E2 mole fraction of around 0.45, VE was at its minimum. Partial molar volume ${\bar{V}}_1$ increases monotonously with the mole fraction x1(=x) and ${\bar{V}}_2$ decreases with x. Any particular point in ${\bar{V}}_1$ and ${\bar{V}}_2$, which may point to molecular association, was unobserved.

Using Numerical Maps to Select Solar Panel Installation Sites no Expressway Slopes (수치지도를 이용한 고속국도 주변 태양광 패널 설치 대상지 선정)

  • Jung, Jaehoon;Kim, Byungil
    • Korean Journal of Construction Engineering and Management
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    • v.17 no.5
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    • pp.71-77
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    • 2016
  • Solar energy is a viable source to replace fossil fuels. However, challenges associated with site selection for solar panel installation inhibit the uptake of solar energy systems. Expressway slopes offer a potentially attractive alternative for solar panel installation for the following reasons: expressway slopes are vacant public sites, they are abundant (about 4,193km in South Korea), and they are linear in nature. Traditoinally when selecting sites for solar systems conventional surveying methods are employed. Unfortunately, these methods can be dangerous, time consuming, and labor intensive. To overcome these limitations of conventional site selection methodologies, we propose an automated approach using numerical maps. First, contour and expressway polylines are extracted separately from numeric maps. The extracted contour lines are then converted into a digital terrain model; this is used to calculate aspect and slope information. Next, the extracted expressway lines are projected onto a binary image and refined to recover the disconnections, and then applied to create a buffer zone to narrow the search space. Finally, all data sets are overlaid to identify candidate sites for solar panel systems and are visually verified through comparisons with aerial photos.

Experiment and Simulation of 2-bed PSA for Hydrogen Separation from H2/CH4 Gas Mixture (수소/메탄 혼합 기체로부터 수소 분리를 위한 두 탑 PSA 실험과 전산 모사)

  • Nam, Gi-Moon;Jeong, Byung-Man;Kang, Seok-Hyun;Lee, Chang-Ha;Lee, Byung-Kwon;Choi, Dae-Ki
    • Korean Chemical Engineering Research
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    • v.43 no.2
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    • pp.249-258
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    • 2005
  • A two-column six-step pressure swing adsorption(PSA) process was to study separation of hydrogen from hydrogen and methane($60_{vol}%/40_{vol}%$) binary system onto activated carbon adsorbent. The effects of the feed gas pressure, the feed flowrate and the P/F(purge to feed) ratio on the process performance were evaluated. The cyclic steady-states of PSA process were reached to after 15 cycles. $H_2$ purity increases according as the P/F ratio and pressure increase and the feed flow rate decreases; however, $H_2$ recovery shows an opposite phenomena to the purity. PSA process simulation studied to find optimum operation condition. In the results, 22 LPM feed flowrate, 11 atm adsorption pressure and 0.10 P/F ratio might be optimal values to obtain more than 75% recovery and 99% purity hydrogen. In this study was non-isothermal and non-adiabatic model considering linear driving force(LDF) model and Langmuir-Freundlich adsorption isotherm considered to compare between prediction and experimental data.

Spectrophotometric Study of Acidity and Complex Formation of Anti-Inflammatory Drug Piroxicam with Some Transition Metal Ions in Different Methanol/Water Mixtures by Chemometric Methods (Chemometric 방법에 의한 메탄올/물 계에서 전이 금속 이온과 소염제 Piroxicam의 산성도 및 착체 형성에 관한 분광광도법 연구)

  • Ghasemi, Jahan B.;Jalalvand, Alireza
    • Journal of the Korean Chemical Society
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    • v.53 no.6
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    • pp.693-703
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    • 2009
  • The complex formation of anti-inflamatory drug piroxicam (PX, 4-hydroxy-2-methyl-N-2--pridyl-2H-1,2-benzothiazine-3-carboxadiamide-1,1-dioxide) with transition metal ions Co(II), Ni(II), Cu(II) and Zn(II) in methanol(MeOH)/water binary mixtures were studied by spectrophotometric method at 25$^{\circ}C$, constant pH = 5.0 and I = 0.1 M. The computer program SQUAD was used to extract the desired information from the spectral data. The outputs of the fitting processes were stability constants, standard deviations of the estimated stability constants, concentration distribution diagrams and spectral profiles of all species. The sequence of the stability constants of PX complexes with Co(II), Ni(II), Cu(II) and Zn(II) follow the Cu(II) > Co(II) > Ni(II) ${\approx}$ Zn(II) order. This may be due to different geometry tendencies of these metal ions. The acidity constants of the PX were also determined under above condition from its absorption spectra at different pH values. The computer program DATAN was used for determination of acidity constants of PX. The validity of the obtained acidity constants was checked by a well known computer program SPECFIT/32. The effects of the different parameters like solvent nature, cations characteristics on the stability and acidity constants were thoroughly discussed.

A study on the growth mechanism of Burger King based on dynamic models of success and failure of businesses

  • Lee, Sang-Youn
    • East Asian Journal of Business Economics (EAJBE)
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    • v.5 no.4
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    • pp.39-49
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    • 2017
  • Purpose - This study is to propose a creative idea for constant business growth and development by examining characteristics of business outcomes by phase, which are "growth" and "erosion and stagnation," respectively. Research design, data, methodology - It is necessary to identify an occurrence of crisis and its diffusion with a dynamic model in order to identify a success and failure of businesses in an organic way, not on a binary structure. The static perspective is to understand a crisis as a simply one-time event or as a linear causation. Thus, it has a limited understanding of the overall situation and has limits to investigating a foundational cause and developing long-term countermeasures. On the contrary, the dynamic perspective is to understand the crisis as circulation process of the overall system. Thus, it divides elements of the crisis as external and internal ones to understand it as the causal relationship of each element. Results - During the growth period of Burger King, the company promoted its brand very successfully with aggressive and creative marketing activities. However, due to the founder's disposal of management rights and the following changes in the management, the company had no choice but to lose focus on its business philosophy and brand management, and eventually it had to face the big crisis (resonance) which was delisting from the stock market because of the external threat; well-being trend. However, Burger King resumed lifting on the stock exchange by making great efforts to clearly identify the current issues and seek solutions. Under the spirit of "perseverance" and its slogan "Have it your way" the company is now going head to head with McDonald's in the North American region and emerging countries. Conclusions - Then, what is the most crucial factor in the success and failure of businesses? Answers may vary, however, as learned from the case study of Burger King, corporations should inspect the present and focus on developing a long-term strategy for the future and actively fulfill the actions. McDonald's may not be able to innovate by itself in the future as it may become routinized to the growth. There will be chances of winning if we change conditions of individuals or organizations to an organic system in terms of being creative. There is a hopeful message here that an individual or small business may have more advantages in the era of the idea and innovation.