• Title/Summary/Keyword: Direct detection

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Real-Time Monitoring of Mitochondrial ATP Synthesis and Hydrolysis by Surface Infrared Spectroscopy

  • Yamaguchi, Ryo-Taro;Hirano-Iwata, Ayumi;Aonuma, Yuki;Yoshimura, Yuya;Shinohara, Yasuo;Kimura, Yasuo;Niwano, Michio
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.02a
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    • pp.108-109
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    • 2013
  • Mitochondria play key roles in the production of cell's energy. Their dominant function is the synthesis of adenosine 5'-triphosphate (ATP) from adenosine diphosphate (ADP) and phosphate (Pi) through the oxidative phosphorylation. Evaluation of drug-induced mitochondrial toxicity has become increasingly important since mitochondrial dysfunction has recently been implicated in numerous diseases including cancer and diabetes mellitus. Mitochondrial functions have been monitored via oxygen consumption, mitochondrial membrane potential, and more importantly via ATP synthesis since ATP synthesis is the most essential function of mitochondria. Various analytical methods have been employed to investigate ATP synthesis in mitochondria, including high performance liquid chromatography (HPLC), bioluminescence technique, and pH measurement. However, most of these methods are based on destructive analysis or indirect monitoring through the enzymatic reaction. Infrared absorption spectroscopy (IRAS) is one of the useful techniques for real-time, label-free, and direct monitoring of biological reactions [1,2]. However, the strong water absorption requires very short path length in the order of several micrometers. Transmission measurements with thin path length are not suitable for mitochondrial assays because solution handlings necessary for evaluating mitochondrial toxicity, such as rapid mixing of drugs and oxygen supply, are difficult in such a narrow space. On the other hand, IRAS in the multiple internal reflection (MIR) geometry provides an ideal optical configuration to combine solution handling and aqueous-phase measurement. We have recently reportedon a real-time monitoring of drug-induced necrotic and apoptotic cell death using MIR-IRAS [3,4]. Clear discrimination between viable and damaged cells has been demonstrated, showing a promise as a label-free and real-time detection for cell-based assays. In the present study, we have applied our MIR-IRAS system to mitochondria-based assays by monitoring ATP synthesis in isolated mitochondria from rat livers. Mitochondrial ATP synthesis and hydrolysis were in situ monitored with MIR-IRAS, while dissolved oxygen level and solution pH were simultaneously monitored with O2 and pH electrodes, respectively. It is demonstrated that ATP synthesis and hydrolysis can be monitored by the IR spectral changes in phosphate groups in adenine nucleotides and MIR-IRAS is useful for evaluating time-dependent drug effects of mitochondrial toxicants.

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An Enzyme-Linked Immunosorbent Assay for $Aflatoxin\;M_1$ in Cow's Milk without a Cleanup Procedure (희석에 의한 우유 중 $Aflatoxin\;M_1$의 효소면역측정법)

  • Shon, Dong-Hwa;Lim, Sun-Hee;Lee, Yin-Won
    • Korean Journal of Food Science and Technology
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    • v.28 no.6
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    • pp.1184-1187
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    • 1996
  • A simple and rapid detection system for $aflatoxin\;M_1\;(AFM_1)$ in cow's milk by an enzyme-linked immunosorbent assay (ELISA) was developed. Specific antibodies against $AFM_1$, conjugated to bovine serum albumin $(AFM_1-BSA)$ were raised in rabbits and purified. The cross-reactivities of the antibodies against aflatoxin analogs were less than 29.9%. When a competitive direct ELISA (cdELISA) for $AFM_1$, established by use of the antibodies was applied to the spike test of $AFM_1$ onto uncontaminated cow's milk, the assay recovery was unstable unless cow's milk was diluted to 40% (2:3) with phosphate buffered saline (PBS). In that condition of sample dilution, the mean ELISA recovery of $AFM_1$, from the cow's milk was 113% (coefficient of variation (CV) of each recovery percentage, 8.2%) in the range of $0.3{\sim}3.0\;ppb$. These results showed that the ELISA system could be a convenient tool to monitor the contamination of AFM1 more than 0.5 ppb in cow's milk (FDA allowance limit) easily.

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Multi-point Dynamic Displacement Measurements of Structures Using Digital Image Correlation Technique (Digital Image Correlation기법을 이용한 구조물의 다중 동적변위응답 측정)

  • Kim, Sung-Wan;Kim, Nam-Sik
    • Journal of the Earthquake Engineering Society of Korea
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    • v.13 no.3
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    • pp.11-19
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    • 2009
  • Recently, concerns relating to the maintenance of large structures have been increased. In addition, the number of large structures that need to be evaluated for their structural safety due to natural disasters and structural deterioration has been rapidly increasing. It is common for the structural characteristics of an older large structure to differ from the characteristics in the initial design stage, and changes in dynamic characteristics may result from a reduction in stiffness due to cracks on the materials. The process of deterioration of such structures enables the detection of damaged locations, as well as a quantitative evaluation. One of the typical measuring instruments used for the monitoring of bridges and buildings is the dynamic measurement system. Conventional dynamic measurement systems require considerable cabling to facilitate a direct connection between sensor and DAQ logger. For this reason, a method of measuring structural responses from a remote distance without the mounted sensors is needed. In terms of non-contact methods that are applicable to dynamic response measurement, the methods using the doppler effect of a laser or a GPS are commonly used. However, such methods could not be generally applied to bridge structures because of their costs and inaccuracies. Alternatively, a method using a visual image can be economical as well as feasible for measuring vibration signals of inaccessible bridge structures and extracting their dynamic characteristics. Many studies have been conducted using camera visual signals instead of conventional mounted sensors. However, these studies have been focused on measuring displacement response by an image processing technique after recording a position of the target mounted on the structure, in which the number of measurement targets may be limited. Therefore, in this study, a model experiment was carried out to verify the measurement algorithm for measuring multi-point displacement responses by using a DIC (Digital Image Correlation) technique.

Space-Time Concatenated Convolutional and Differential Codes with Interference Suppression for DS-CDMA Systems (간섭 억제된 DS-CDMA 시스템에서의 시공간 직렬 연쇄 컨볼루션 차등 부호 기법)

  • Yang, Ha-Yeong;Sin, Min-Ho;Song, Hong-Yeop;Hong, Dae-Sik;Gang, Chang-Eon
    • Journal of the Institute of Electronics Engineers of Korea TC
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    • v.39 no.1
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    • pp.1-10
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    • 2002
  • A space-time concatenated convolutional and differential coding scheme is employed in a multiuser direct-sequence code-division multiple-access(DS-CDMA) system. The system consists of single-user detectors (SUD), which are used to suppress multiple-access interference(MAI) with no requirement of other users' spreading codes, timing, or phase information. The space-time differential code, treated as a convolutional code of code rate 1 and memory 1, does not sacrifice the coding efficiency and has the least number of states. In addition, it brings a diversity gain through the space-time processing with a simple decoding process. The iterative process exchanges information between the differential decoder and the convolutional decoder. Numerical results show that this space-time concatenated coding scheme provides better performance and more flexibility than conventional convolutional codes in DS-CDMA systems, even in the sense of similar complexity Further study shows that the performance of this coding scheme applying to DS-CDMA systems with SUDs improves by increasing the processing gain or the number of taps of the interference suppression filter, and degrades for higher near-far interfering power or additional near-far interfering users.

Analysis of Nitrosamines Concentration in Condom by using LC-MS/MS (LC-MS/MS를 이용한 콘돔에 함유된 니트로사민류 농도 분석)

  • Park, Na-Youn;Kim, Sungmin;Jung, Woong;Kho, Younglim
    • Journal of the Korean Chemical Society
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    • v.62 no.3
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    • pp.181-186
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    • 2018
  • Nitrosamines are the nitrosocompounds which are produced by nitrosation reactions of the secondary amine and nitrite, and has been found to be produced through the vulcanization process during the production of rubber products Recently, nitrosamines have been detected in rubber products and become a major topic. Condoms are disposable medical devices, so safety is important because they come into direct contact with the skin and mucous membranes. In this study, we developed an analytical method for nitrosamines in condoms by applying ISO 29941 method. The samples were eluted by distilled water, and target compounds were extracted by liquid-liquid extraction with dichloromethane. And then after concentrated, and quantitatively analyzed by LC-MS/MS. The accuracies of the analytical method were ranged from 85.8 to 108.7%, precisions were lower than 11.5%, and the detection limits were from 0.11 (NDPA and NDBA) to 0.48 (NPYR) ng/mL. Among the 31 condom samples, NDBA was detected from 2 cases by extraction of distilled water, and NDMA were detected from 1 case, NDEA from 4 cases and NDBA from 26 cases by extraction of artificial saliva (pH 4.5). The total amount of nitrosamines in all samples were less than $500{\mu}g/kg$.

Effects of Joint Mobilization Techniques on the Joint Receptors (관절 가동운동이 관절 감수기에 미치는 영향)

  • Kim, Suhn-Yeop
    • The Journal of Korean Academy of Orthopedic Manual Physical Therapy
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    • v.2 no.1
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    • pp.9-19
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    • 1996
  • Type I, II, III are regarded as "true" joint receptors, type IV is considered a class of pain receptor. Type I, II and III mechanoreceptors, via static and dynamic input, signal joint position, intraarticular pressure changes, and the direction, amplitude, and velocity of joint movements. Type I mechanoreceptor subserve both static and dynamic physiologic functions. Type I are found primarily in the stratum fibrosum of the joint capsule and ligaments. Type I receptors have a low threshold for activation and are allow to adapt to changes altering their firing frequency. Type II receptors have a low threshold for activation. These dynamic receptors respond to joint movement. Type II receptors are thus termed rapidly adapting. Type II joint receptors are located at the junction of the synovial membrane and fibrosum of the joint capsule and intraarticular and extraarticular fat pads. Type III receptors have been found in collateral ligaments of the joints of the extremities. Morphologically similar to Golgi tendon organ. These dynamic receptors have a high threshold to stimulation and are slowly adating. Type IV receptors possess free nerve ending that have been found in joint capsule and fat pads. They are not normally active, but respond to extreme mechanical deformation of the joint as well as to direct chemical or mechanical irritation. Small amplitude oscillatory and distraction movements(joint mobilization) techniques are used to stimulate the mechanoreceptors that may inhibit the transmission of nociceptors stimuli at the spinal cord or brain stem levels.

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BACTERIAL IDENTIFICATION WITH RANDOM-CLONED RESTRICTION FRAGMENT OF Porphyromonas endodontalis ATCC 35406 GENOMIC DNA (무작위로 클로닝한 Porphyromonas endodontalis ATCC 35406 지놈 DNA의 제한절편 hybridization법에 의한 세균동정)

  • Um, Won-Seok;Han, Yoon-Soo
    • Restorative Dentistry and Endodontics
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    • v.20 no.2
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    • pp.645-654
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    • 1995
  • Porphyromonas endodontalis is a black-pigmented anaerobic Gram negative rod which is associated with endodontal infections. It has been isolated from infected dental root canals and submucous abscesses of endodontal origin. DNA probe is an available alternative, offering the direct detection of a specific microorganism. Nucleic-acid probes can be off different types: whole different: whole-genomic, cloned or oligonucleotide probes. Wholegenomic probes are the most sensitive because the entire genome is used for possible hybridization sites. However, as genetically similar species of bacteria are likely to be present in specimences, cross-reactions need to be considered. Cloned probes are isolated sequences of DNA that do not show cross-reactivity and are produced in quantity by cloning in a plasmid vector. Cloned probes can approach the sensitivity found with whole-genomic probes while avoiding known cross-reacting species. Porphyromonas endodontalis ATCC 35406 (serotype $O_1K_1$) was selected in this experiment to develop specific cloned DNA probes. EcoR I-digested genomic DNA fragments of P. endodontalis ATCC 35406 were cloned into pUC18 plasmid vector. From the E. coli transformed with the recombinant plasmid 4 clones were selected to be tested as specific DNA probes. Restriction-digested whole-genomic DNAs prepared from P. gingivalis 38(serotype a), W50(serotype b), A7A1-28(serotype C), P. intermedia 9336(serotype b), G8-9K-3(serotype C), P. endodontalis ATCC 35406(serotype $O_1K_1$), A. a Y4(serotype b), 75(serotype a), 67(serotype c), were each seperated on agarose gel electrophoresis, blotted on nylon membranes, and were hybridized with digoxigenin-dUTP labeled probe. The results were as follows: 1. Three clones of 1.6kb(probe e), 1.6kb(probe f), and 0.9kb(probe h) in size, were obtained. These clones were identified to be a part of the genomic DNA of P. endodontalis ATCC 35406 judging from their specific hybridization to the genomic DNA fragments of their own size on Southern blot. 2. The clones of 4.9kb(probe i) was identified to be a part of the genomic DNA of P. endodontalis ATCC 35406. but not to specific for itself. It was hybridized to P. gingivalis A7A1-28, P. intermedia G89K-3.

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The study of the stereo X-ray system for automated X-ray inspection system using 3D-reconstruction shape information (3차원 형상복원 정보 기반의 검색 자동화를 위한 스테레오 X-선 검색장치에 관한 연구)

  • Hwang, Young-Gwan;Lee, Nam-Ho
    • Journal of the Korea Institute of Information and Communication Engineering
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    • v.18 no.8
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    • pp.2043-2050
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    • 2014
  • As most the scanning systems developed until now provide radiation scan plane images of the inspected objects, there has been a limitation in judging exactly the shape of the objects inside a logistics container exactly with only 2-D radiation image information. As a radiation image is just the density information of the scanned object, the direct application of general stereo image processing techniques is inefficient. So we propose that a new volume-based 3-D reconstruction algorithm. Experimental results show the proposed new volume based reconstruction technique can provide more efficient visualization for X-ray inspection. For validation of the proposed shape reconstruction algorithm using volume, 15 samples were scanned and reconstructed to restore the shape using an X-ray stereo inspection system. Reconstruction results of the objects show a high degree of accuracy compared to the width (2.56%), height (6.15%) and depth (7.12%) of the measured value for a real object respectively. In addition, using a K-Mean clustering algorithm a detection efficiency of 97% is achieved. The results of the reconstructed shape information using the volume based shape reconstruction algorithm provide the depth information of the inspected object with stereo X-ray inspection. Depth information used as an identifier for an automated search is possible and additional studies will proceed to retrieve an X-ray inspection system that can greatly improve the efficiency of an inspection.

Fabrication and Characterization of Lead Oxide (PbO) Film for High Efficiency X-ray Detector (고효율 X선 검출기 적용을 위한 PbO 필름 제작 및 특성 연구)

  • Cho, Sung-Ho;Kang, Sang-Sik;Choi, Chi-Won;Kwun, Chul;Nam, Sang-Hee
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2007.06a
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    • pp.329-329
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    • 2007
  • Photoconductive poly crystalline lead oxide coated on amorphous thin film transistor (TFT) arrays is the best candidate for direct digital x-ray detector for medical imaging. Thicker films with lessening density often show lower x-ray induced charge generation and collection becomes less efficient. In this work, we present a new methodology used for the high density deposition of PbO. We investigate the structural properties of the films using X-ray diffraction and electron microscopy experiments. The film coatings of approximately $200\;{\mu}m$ thickness were deposited on $2"{\times}2"$ conductive-coated glass substrates for measurements of dark current and x-ray sensitivity. The lead oxide (PbO) films of $200\;{\mu}m$ thickness were deposited on glass substrates using a wet coating process in room temperature. The influence of post-deposition annealing on the characteristics of the lead oxide films was investigated in detail. X-ray diffraction and scanning electron microscopy, and atomic force microscopy have been employed to obtain information on the morphology and crystallization of the films. Also we measured dark current, x-ray sensitivity and linearity for investigation of the electrical characteristics of films. It was found that the annealing conditions strongly affect the electrical properties of the films. The x-ray induced output charges of films annealed in oxygen gas increases dramatically with increasing annealing temperatures up to $500^{\circ}C$ but then drops for higher temperature anneals. Consequently, the more we increase the annealing temperatures, the better density and film quality of the lead oxide. Analysis of this data suggests that incorporation and decomposition reactions of oxygen can be controlled to change the detection properties of the lead oxide film significantly. Post-deposition thermal annealing is also used for densely film. The PbO films that are grown by new methodology exhibit good morphology of high density structure and provide less than $10\;pA/mm^2$ dark currents as they show saturation in gain (at approximate fields of $4\;V/{\mu}m$). The ability to operate at low voltage gives adequate dark currents for most applications and allows voltage electronics designs.

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Development of an Enzyme-Linked Immunosorbent Assay Method for Residual Host Cell derived Proteins in Recombinant Antibody Drug Production (재조합 항체의약품의 생산시 생산세포주 유래 단백질 검출을 위한 ELISA 방법 개발)

  • Joung, Chan-Hi;Lim, Sang-Min;Koo, Yoon-Mo;Lee, Yong-Yoon;Son, Young-Su;Kim, Hyun-Il;Park, Heung-Rok
    • KSBB Journal
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    • v.21 no.3
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    • pp.212-219
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    • 2006
  • The purpose of this study was to develop a assay system of host cell-derived residual proteins in final pharmaceutical products. Accurate and simple assay system for host cell-derived proteins(HCPs) is very important test item in pharmaceutical qualification control. In this study, methods for quantification of residual HCPs in recombinant anti-GPIIbIIIa antibody were developed using a process-specific immunoligand assay which was based on the Enzyme linked Immunosorbent assay(ELISA) system. The assay had a detection limit of 10.8 ng/ml of HCPs with a product concentration of 1 mg/ml. The practical implication of these results is that the developed ELISA system can be used for HCPs qualification control and this system will be applicable to develop another ELISA system of different antibody drug.