• Title/Summary/Keyword: Sensitivity derivative

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Development and Application of Enzyme Immunoassay for Endosulfan Residue Analysis (Endosulfan 과 그 분해산물의 Enzyme Immunoassay에 의한 분석법의 개발과 응용)

  • Suh, Yong-Tack;Shim, Jae-Han;Lee, Kang-Bong
    • Korean Journal of Environmental Agriculture
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    • v.11 no.1
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    • pp.59-66
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    • 1992
  • An enzyme immunoassay(EIA) was developed for the analysis of insecticide endosulfan and its degradation products. The sensitivity and specificity of the antibody produced were examined. Optimal conditions in the ELISA system for residue analysis were also discussed. A mixed suspension of endosulfan-hemocyanin conjugate(ES-KLH) 1.1 mg/ml and Freund's adjuvant was injected subcutaneously to white rabbits and then collected antisera were tested for titers by indirect ELISA(1/24,000). Because of difficulties in the synthesis of endosulfan peroxidase conjugate, amine derivative of endosulfan-diol was synthesized and it showed 40% of conjugate yields(2mg/ml of conjugate). the highest sensitivity obtained enzyme-conjugate was a concentration of 200ng/ml. The detection limit of endosulfan in ELISA system was 5 ppb on the standard curve. In application of ELISA for residue analysis, the recoveries were really 100% both in the spiked soil and apple sample regardless of endosulfan concentration treated. On the other hand, chlorinated hydrocarbons of similar structure with endosulfan showed low cross-reactivity$(2.2%{\sim}29.2%).$

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Inhibition of Barley Acetolactate Synthase by Triazolopyrimidine Derivative (트리아졸로피리미딘계 유도체에 의한 보리 Acetolactate Synthase의 저해)

  • Kim, Sung Ho;NamGoong, Sung Keon;Shin, Jung Hyu;Chang, Soo Ik;Choi, Jung Do
    • Journal of the Korean Chemical Society
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    • v.43 no.4
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    • pp.461-468
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    • 1999
  • Acetolactate synthase (ALS) catalyzes the first common reaction in the biosynthesis of branched-chain amino acids, valine, leucine, and isoleucine. ALS is the common target of several classes of structurally diverse herbicides, the triazolopyrimidines, the imidazolinones, the sulfonylureas, and pyrimidyl-oxy-benzoates. We examined ihibitory activities of newly synthesized triazolopyrimidine sulfonamide derivatives using partially purified ALS from barley. $IC_{50}$ values for the active derivatives are 0.5nM∼8$\mu$M. Among them TP1 and TP2 are the most potent ALS inhibitors with $IC_{50}$ values of 0.5nM and 1.6nM, respectively. These inhibitors are more potent in the inhibition of barley ALS than commercial herbicides, Metosulam ($IC_{50}$;3.6 nM), Flumetsulam ($IC_{50}$;126 nM), and Cadre ($IC_{50};4 {\mu}M$). The progress curves for inhibition of ALS by TP2 showed that the amount of inhibition increases with time. The inhibition of ALS by TP2 was mixed-type inhibition with respect to pyruvate. Dual inhibition analyses of TP2 versus an imidazolinone, Cadre, and Leu showed parallel and intercepting kinetic pattern, respectively. The results suggest that TP2 binds to ALS competively with Cadre but not with Leu. Chemical modification of cysteinly residues in ALS decreased the sensitivity of ALS to Leu, while the modification did not affect the sensitivity of ALS to TP2 and Cadre.

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Determination of homogentisic acid in human plasma by GC-MS for diagnosis of alkaptonuria (GC-MS를 이용한 혈장 중 호모겐티식산의 분석;알캅톤뇨증의 진단)

  • Thapa, Maheshwor;Yu, Jundong;Lee, Wonjae;Islam, Fokhrul;Yoon, Hye-Ran
    • Analytical Science and Technology
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    • v.28 no.5
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    • pp.323-330
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    • 2015
  • Alkaptonuria, a rare inherited metabolic disease, is characterized by a lack of homogentisate dioxygenase and accumulation of homogentisic acid (HGA), leading to homogentisic aciduria, arthritis, and ochronosis. In this study, a rapid analytical method, without an expensive and tedious solid phase extraction step, was developed to quantify HGA in plasma using GC-MS. HGA-spiked pooled plasma samples were subjected to liquid-liquid extraction (LLE) with ethyl acetate, followed by trimethylsilyl derivatization (TMS) and GC-MS quantification using selected ion monitoring. The formation of TMS derivative of the 1 carboxylic and 2 hydroxyl functional groups was performed by reacting BSTFA (with 10% TMCS) for 5 min at 80 ℃. For selected ion monitoring, quantification and confirmation ions were determined based on specific ions (m/z 384, m/z 341 and m/z 252) of the TMS derivative of HGA. Calibration curves of pooled normal plasma specimens showed a linear relationship in the range of 1-100 ng/µL. The precision and accuracy were within a relative standard deviation (RSD) of 1 to 15% and a bias of -5 to 25%. Recoveries were obtained in the range of 99-125% and 95-115% for intra-day and inter-day assay, respectively, at 2, 20 and 80 ng/µL. The limit of detection (LOD) and limit of quantification (LOQ) were 0.4 ng/µL and 4 ng/µL, respectively. No homogentisic acid was excreted from normal Korean plasma samples. Collectively, the results from the present study suggest that this method could be useful for routine diagnosis and therapeutic monitoring of alkaptonuria patients with excellent sensitivity and rapidity.

A Novel Chenodeoxycholic Derivative HS-1200 Enhances Radiation-induced Apoptosis in Human MCF-7 Breast Cancer Cells (담즙산 합성유도체(HS-1200)가 인체 유방암 세포주(MCF-7)에서 유도하는 방사선 감작 효과)

  • Lee Hyung Sik;Choi Young Min;Kwon Hyuk Chan;Song Yeon Suk
    • Radiation Oncology Journal
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    • v.22 no.2
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    • pp.145-154
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    • 2004
  • Purpose : To examine whether a synthetic bile acid derivatives (HS-1200) sensitizes the radiation-induced apoptosis in human breast cancer cells (MCF-7) and to investigate the underlying mechanism. Materials and Methods : Human breast cancer cells (MCF-7) in exponential growth phase were treated with HS-1200 for 24 hours at 37$^{\circ}C$ with 5$\%$ CO$_{2}$ in air atmosphere. After removal of HS-1200, cells were irradiated with 2$\~$8 Gy X-ray, and then cultured Ii drug-free media for 24-96 hours. The effect of radiation on the clonogenicity of MCF-7 cells was determined with clonogenic cell survival assay with 16$\mu$M of HS-1200. The induction of apoptosis was determined using agarose gel electrophoresis and Hoechst staining. The expression level of apoptosis-related molecules, such as PARP, Bax, Bcl-2, Bak and AIF, were assayed by Western blotting analysis with 40$\mu$M of HS-1200 combined with 8 Gy irradiation. To examine the cellular location of cytochrome c, bax and AIF immunofluorescent stainings were undertaken. Results : Treatment of MCF-7 cells with 40$\mu$M of HS-1200 combined with 8 Gy irradiation showed several changes associated with enhanced apoptosis by agarose gel electrophoresis and Hoechst staining. HS-1200 combined with 8 Gy irradiation treatment also enhanced production of PARP cleavage products and increased Bax/Bcl-2 ratio by Western blotting. Loss of mitochondrial membrane potential ($\Delta$$\psi$$_{m}$) and increased cytochrome c staining indicated that cytochrome c had been released from the mitochondria in HS-1200 treated cells. Conclusion : We demonstrated that combination treatment with a synthetic chenodeoxycholic acid derivative HS-1200 and irradiation enhanced radiation-induced apoptosis of human breast cancer cells (MCF-7). We suggest that the increased Bax/Bcl-2 ratio In HS-1200 co-treatment group underlies the increased radio sensitivity of MCF-7 cells. Further futures studies are remained elusive.

Development of a Model for Calculating Road Congestion Toll with Sensitivity Analysis (민감도 분석을 이용한 도로 혼잡통행료 산정 모형 개발)

  • Kim, Byung-Kwan;Lim, Yong-Taek;Lim, Kang-Won
    • Journal of Korean Society of Transportation
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    • v.22 no.5
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    • pp.139-149
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    • 2004
  • As the expansion of road capacity has become impractical in many urban areas, congestion pricing has been widely considered as an effective method to reduce urban traffic congestion in recent years. The principal reason is that the congestion pricing may lead the user equilibrium (UE) flow pattern to system optimum (SO) pattern in road network. In the context of network equilibrium, the link tolls according to the marginal cost pricing principle can user an UE flow to a SO pattern. Thus, the pricing method offers an efficient tool for moving toward system optimal traffic conditions on the network. This paper proposes a continuous network design program (CNDP) in network equilibrium condition, in order to find optimal congestion toll for maximizing net economic benefit (NEB). The model could be formulated as a bi-level program with continuous variable(congestion toll) such that the upper level problem is for maximizing the NEB in elastic demand, while the lower level is for describing route choice of road users. The bi-level CNDP is intrinsically nonlinear, non-convex, and hence it might be difficult to solve. So, we suggest a heuristic solution algorithm, which adopt derivative information of link flow with respect to design parameter, or congestion toll. Two example networks are used for test of the model proposed in the paper.

Effects of cGMP on the Contractility and Ca Movement in the Aorta of Normotensive Wistar-Kyoto Rats and Spontaneously Hypertensive Rats

  • Park, Hae-Kun;Jeon, Byeong-Hwa;Kim, Se-Hoon;Kim, Hoe-Suk;Chang, Seok-Jong
    • The Korean Journal of Physiology
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    • v.28 no.2
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    • pp.181-190
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    • 1994
  • Endothelium-derived relaxing factor (EDRF) activates guanylate cyclase which mediates the formation of cGMP from GTP in vascular smooth muscle. It is well known that endothelium-dependent relaxation is impaired in spontaneously hypertensive rats (SHR). However, it is still unknown whether the impaired endothelium-dependent relaxation in SHR results from the reduced release of EDRF or from the decrease of vascular response to EDRF. We investigated the effects of cGMP on the contractility and Ca movement in the aorta of SHR and Wistar-Kyoto rats (WKY). The amplitude of the endothelium-dependent relaxation to actylcholine (ACh) was significantly less in SHR than in WKY. L-arginine $(10^{-3}M)$ did not increase endothelium-dependent relaxation in both strains. Sodium nitroprusside (SNP), an activator of guanylate cyclase, relaxed the 40 mM $K^+-induced$ contraction in a dose-dependent manner $(10^{-10}{\sim}10^{-6}\;M)$ in the endothelium-rubbed aortic strips of both strains. However, there was no significant difference in these relaxations between WKY and SHR. 8-bromo-cyclic guanosine monophosphate (8-Br-cGMP), a cell membrane-permeable derivative of cGMP relaxed the 40 mM $K^+-induced$ contraction in a dose-dependent manner $(10^{-6}{\sim}10^{-4}\;M)$ in the endothelium-rubbed aortic strips of both strains. Also norepinephrine $(10^{-6}\;M)-induced$ contractions in normal and Ca-free Tyrode's solution were suppressed by the pretreatment with 8-Br-cGMP $(10^{-4}\;M)$ in either strain. However, the amplitudes of suppression induced by 8-Br-cGMP were greater in SHR than that in WKY. Basal $^{45}Ca$ uptake and 40mM $K^+-stimulated\;^{45}Ca$ uptake were not suppressed by pretreatment with 8-Br-cGMP $(10^{-4}\;M)$ in single aortic smooth muscle cells of both SHR and WKY. From the above results, it is suggested that cGMP decreases Ca sensitivity in vascular smooth muscle cells and that the impaired endothelium-dependent relaxation in the aortic strips of SHR is not the result of a reduced vascular response to EDRF.

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Genome-wide Drug-induced Haploinsufficiency Screening of Fission Yeast for Identification of Hydrazinocurcumin Targets

  • Baek, Seung-Tae;Kim, Dong-Uk;Han, Sang-Jo;Woo, Im-Sun;Nam, Mi-Young;Kim, Li-La;Heo, Kyung-Sun;Lee, Hye-Mi;Hwang, Hye-Rim;Choi, Shin-Jung;Won, Mi-Sun;Lee, Min-Ho;Park, Song-Kyu;Lee, Sung-Hou;Kwon, Ho-Jeong;Maeng, Pil-Jae;Park, Hee-Moon;Park, Young-Woo;Kim, Dong-Sup;Hoe, Kwang-Lae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.263-269
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    • 2008
  • Hydrazinocurcumin (HC), a synthetic derivative of curcumin, has been reported to inhibit angiogenesis via unknown mechanisms. Understanding the molecular mechanisms of the drug's action is important for the development of improved compounds with better pharmacological properties. A genome-wide drug-induced haploinsufficiency screening of fission yeast gene deletion mutants has been applied to identify drug targets of HC. As a first step, the 50% inhibition concentration $(IC_{50})$ of HC was determined to be $2.2{\mu}M$. The initial screening of 4,158 mutants in 384-well plates using robotics was performed at concentrations of 2, 3, and $4{\mu}M$. A second screening was performed to detect sensitivity to HC on the plates. The first screening revealed 178 candidates, and the second screening resulted in 13 candidates, following the elimination of 165 false positives. Final filtering of the condition-dependent haploinsufficient genes gave eight target genes. Analysis of the specific targets of HC has shown that they are related to septum formation and the general transcription processes, which may be related to histone acetyltransferase. The target mutants showed 65% growth inhibition in response to HC compared with wild-type controls, as shown by liquid culture assay.

Review on the Three-Dimensional Inversion of Magnetotelluric Date (MT 자료의 3차원 역산 개관)

  • Kim Hee Joon;Nam Myung Jin;Han Nuree;Choi Jihyang;Lee Tae Jong;Song Yoonho;Suh Jung Hee
    • Geophysics and Geophysical Exploration
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    • v.7 no.3
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    • pp.207-212
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    • 2004
  • This article reviews recent developments in three-dimensional (3-D) magntotelluric (MT) imaging. The inversion of MT data is fundamentally ill-posed, and therefore the resultant solution is non-unique. A regularizing scheme must be involved to reduce the non-uniqueness while retaining certain a priori information in the solution. The standard approach to nonlinear inversion in geophysis has been the Gauss-Newton method, which solves a sequence of linearized inverse problems. When running to convergence, the algorithm minimizes an objective function over the space of models and in the sense produces an optimal solution of the inverse problem. The general usefulness of iterative, linearized inversion algorithms, however is greatly limited in 3-D MT applications by the requirement of computing the Jacobian(partial derivative, sensitivity) matrix of the forward problem. The difficulty may be relaxed using conjugate gradients(CG) methods. A linear CG technique is used to solve each step of Gauss-Newton iterations incompletely, while the method of nonlinear CG is applied directly to the minimization of the objective function. These CG techniques replace computation of jacobian matrix and solution of a large linear system with computations equivalent to only three forward problems per inversion iteration. Consequently, the algorithms are efficient in computational speed and memory requirement, making 3-D inversion feasible.

Development of Enzyme-Protein Binding Assay for Rapid and Sensitive Analysis of Biotin (Biotin 정량분석틀 위한 효소-단백질결합 분석법(EPBA)의 개발)

  • Lee, Kyung-Ae;Shon, Dong-Hwa;Ko, Young-Tae
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1273-1278
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    • 1998
  • Conditions for enzyme-protein binding assay (EPBA) were established in order to detect biotin more rapidly and reproducibly than traditional microbiological assay (MBA). EPBA with streptavidin and biotin-KLH conjugate showed cross-reactivities on biocytin, a derivative with biotin activity, at the rate of 109% $(IC_{50}=0.3\;ppb)\;and\;197%\;(IC_{50}=0.8\;ppb)$, respectively, but not on other derivatives with no biotin activities, such as desthiobiotin, diaminobiotin and 2-iminobiotin. Detection ranges of biotin by EPBA with streptavidin and biotin-KLH conjugates were $0.01{\sim}30\;ng/mL\;and\;0.01{\sim}1.0\;ng/mL(ppb)$, respectively. In the spike test with milk, fruit flake and pine-carrot juice, the correlation coefficience between MBA and EPBA with biotin-KLH conjugates was r=0.994. But MBA showed cross-reactivities both on biocytin and desthiobiotin at the rate of 80.1% and 66.7%, respectively. Detection range of biotin by MBA was $0.1{\sim}0.5\;ng/mL(ppb)$. These results strongly suggest that EPBA is efficient for biotin detection in sensitivity, detection range, cross-reactivity and time consuming.

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Effect of Sodium Butyrate on Blood Glucose, Serum Lipid Profile and Inflammation in Streptozotocin-induced Diabetic Mice (스트렙토조토신으로 유도한 당뇨마우스에서 Sodium Butyrate의 혈당, 혈청 지질 성상 및 염증 억제에 미치는 영향)

  • Yun, Jung-Mi
    • The Korean Journal of Food And Nutrition
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    • v.28 no.2
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    • pp.171-177
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    • 2015
  • Sodium butyrate is a short-chain fatty acid derivative found in foods, such as Parmesan cheese and butter and is produced by anaerobic bacteria fermentation of dietary fibers in the large intestine. There have been reports that butyrate prevented obesity, protected insulin sensitivity, and ameliorated dyslipidemia in dietary obese mice. This study investigated the effects of sodium butyrate on fasting blood glucose level and serum lipid profile in streptozotocin(STZ)-induced diabetic mice. Male C57BL/6 mice were fed AIN-93G for four weeks prior to intraperitoneal injections with STZ (100 mg/kg body weight). Diabetic mice had supplements of 5% sodium butyrate for four weeks. The 5% sodium butyrate diet significantly improved fasting blood glucose level and lipid profile in STZ-induced diabetic mice. Inflammation has been recognized to decrease beta cell insulin secretion and increase insulin resistance. Circulating cytokines can directly affect beta cell function, leading to secretory dysfunction and increased apoptosis. Thus, anti-inflammatory therapies represented a potential approach for the therapy of diabetes and its complications. In this animal study, the 5% sodium butyrate supplementation also inhibited inflammatory cytokine production in STZ-induced diabetic mice. These results suggested that sodium butyrate can be a potential candidate for the prevention of diabetes and its complications.