• Title/Summary/Keyword: Fluorescence polarization

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Transbilayer Effects of Chlorpromazine.HCl on Rotational Mobility of Synaptosomal Plasma Membrane Vesicles Isolated from Bovine Brain

  • Ahn, Ki-Weon;Choi, Chang-Hwa;Kim, Inn-Se;Chung, In-Kyo;Cho, Goon-Jae;Jang, Hye-Ock;Yun, Il
    • BMB Reports
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    • v.33 no.6
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    • pp.541-547
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    • 2000
  • Fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) was used to evaluate the effects of chlorpromazine HCl on the range of the rotational mobility of bulk bilayer structure of the synaptosomal plasma membrane vesicles (SPMV) isolated from a bovine brain. In a dose-dependent manner, chlorpromazine HCl increased the anisotropy (r), limiting anisotropy ($r_{\infty}$) and order parameter (S) of DPH in the membranes. Cationic 1-[4-(trimethylammonio)-phenyl]-6-phenylhexa-1,3,5-hexatriene (TMA-DPH) and anionic 3-[p-(6-phenyl)-1,3,5-hexatrienyl]-phenylpropionic acid (PRO-DPH) were utilized to examine the range of transbilayer asymmetric rotational mobility of the neuronal membranes. The anisotropy (r) of TMA-DPH in the inner monolayer was 0.034 greater than the value of PRO-DPH in the outer monolayer of the membranes. Both cationic TMA-DPH and anionic PRO-DPH were also used to examine the transbilayer asymmetric effects of chlorpromazine HCl on the range of rotational mobility of the membranes. Chlorpromazine HCl have a decreasing effects on the rotational mobility of the bulk bilayer structures and have a greater decreasing effect on the mobility of the inner monolayer as compared to the outer monolayer of the membranes. It has been proven that chlorpromazine HCl exhibit a selective rather than nonselective fluidizing effect within the transbilayer domains of the SPMV.

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Development of a Simultaneous Detection and Quantification Method of Anorectics in Human Urine Using GC-MS and its Application to Legal Cases (GC-MS를 이용한 사람 뇨시료 중 비만치료제 분석 및 비만치료제 남용 현황의 법과학적 고찰)

  • Choi, Hyeyoung;Lee, Jaesin;Jang, Moonhee;Yang, Wonkyung;Kim, Eunmi;Choi, Hwakyung
    • YAKHAK HOEJI
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    • v.57 no.6
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    • pp.420-425
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    • 2013
  • Phentermine (PT) and phenmetrazine (PM) have been widely used as anti-obesity drugs. These drugs should be used with caution due to its close relation to amphetamine in its structure and toxicity. PT and PM, amphetamine-type anorectics, have recently been considered as alternatives for methamphetamine abuse in Korea. In addition, the misuse and abuse of PT and PM obtained by illegal sources such as the internet become a serious social problem. In the present study, a simultaneous detection and quantification method for determining PT and PM in human urine was developed and validated according to the international guidelines. The urine samples were screened using a fluorescence polarization immunooassay and analyzed by gas chromatography mass spectrometry (GC-MS) after extraction using automatic solid phase extraction (SPE) with a mixed-mode cation exchange cartridge and derivatization with pentafluoropropionic anhydride (PFPA). The validation results for selectivity, linearity, limits of detection (LOD) and quantification (LOQ), intra- and inter-assay precision and accuracy and recovery were satisfactory. The validated method was successfully applied to authentic urine samples collected from 38 drug abuse suspects. PT and/or PM were identified with or without methamphetamine in urine samples. Abuse of PT and PM have increased continuously in Korea, therefore, closer supervision of the inappropriate use of anoretics is necessary.

Formation of Calcareous Deposit on Steel Plate by using Waste Oyster Shell (강판상에 굴 패각을 이용한 탄산칼슘 피막의 형성)

  • Kim, Beomsoo;Kwon, Jaesung;Kim, Yeonwon;Lee, Myeonghoon;Yang, Jeonghyeon
    • Journal of the Korean institute of surface engineering
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    • v.50 no.6
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    • pp.531-535
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    • 2017
  • Enormous amount of waste oyster-shell (OS) has a major disposal problem in coastal regions. OSs have attracted much attention for recycling, because these are mainly composed of calcium carbonate with rare impurities. In this study, we demonstrate the calcareous deposit films on steel plate by using OSs on the basic of cathodic protection technique. The composition of the OSs was analyzed by wavelength dispersive X-ray fluorescence spectrometer. Carbon dioxide gas was pumped into distilled water to make carbonic acid solution for dissolution of OS. The calcareous deposit was characterized by second electron microscope (SEM), energy dispersive X-ray analysis (EDX) and X-ray diffraction. Corrosion rates were estimated by measurements of anodic polarization and immersion test. It is confirmed that calcareous deposits on steel plate are formed under all condition of cathodic protection by using waste OS from the SEM and EDX results. Calcareous deposits on steel by OS provide good corrosion resistance by acting as a barrier to oxygen supply to the steel surface.

Investigating the Morphology and Kinetics of Three-Dimensional Neuronal Networks on Electro-Spun Microstructured Scaffolds

  • Kim, Dongyoon;Kim, Seong-Min;Kang, Donghee;Baek, Goeun;Yoon, Myung-Han
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.277.2-277.2
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    • 2013
  • Petri dishes and glass slides have been widely used as general substrates for in vitro mammalian cell cultures due to their culture viability, optical transparency, experimental convenience, and relatively low cost. Despite the aforementioned benefit, however, the flat two-dimensional substrates exhibit limited capability in terms of realistically mimicking cellular polarization, intercellular interaction, and differentiation in the non-physiological culture environment. Here, we report a protocol of culturing embryonic rat hippocampal neurons on the electro-spun polymeric network and the results from examination of neuronal cell behavior and network formation on this culture platform. A combinatorial method of laser-scanning confocal fluorescence microscopy and live-cell imaging technique was employed to track axonal outgrowth and synaptic connectivity of the neuronal cells deposited on this model culture environment. The present microfiber-based scaffold supports the prolonged viability of three-dimensionally-formed neuronal networks and their microscopic geometric parameters (i.e., microfiber diameter) strongly influence the axonal outgrowth and synaptic connection pattern. These results implies that electro-spun fiber scaffolds with fine control over surface chemistry and nano/microscopic geometry may be used as an economic and general platform for three-dimensional mammalian culture systems, particularly, neuronal lineage and other network forming cell lines.

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Diagnostic Accuracy of the Quidel Sofia Rapid Influenza Fluorescent Immunoassay in Patients with Influenza-like Illness: A Systematic Review and Meta-analysis

  • Lee, Jonghoo;Song, Jae-Uk;Kim, Yee Hyung
    • Tuberculosis and Respiratory Diseases
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    • v.84 no.3
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    • pp.226-236
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    • 2021
  • Background: Although the Quidel Sofia rapid influenza fluorescent immunoassay (FIA) is widely used to identify influenza A and B, the diagnostic accuracy of this test remains unclear. Thus, the objective of this study was to determine the diagnostic performance of this test compared to reverse transcriptase-polymerase chain reaction. Methods: A systematic literature search was performed using MEDLINE, EMBASE, and the Cochrane Central Register. Pooled sensitivity, specificity, diagnostic odds ratio (DOR), and a hierarchical summary receiver-operating characteristic curve (HSROC) of this test for identifying influenza A and B were determined using meta-analysis. A sensitivity subgroup analysis was performed to identify potential sources of heterogeneity within selected studies. Results: We identified 17 studies involving 8,334 patients. Pooled sensitivity, specificity, and DOR of the Quidel Sofia rapid influenza FIA for identifying influenza A were 0.78 (95% confidence interval [CI], 0.71-0.83), 0.99 (95% CI, 0.98-0.99), and 251.26 (95% CI, 139.39-452.89), respectively. Pooled sensitivity, specificity, and DOR of this test for identifying influenza B were 0.72 (95% CI, 0.60-0.82), 0.98 (95% CI, 0.96-0.99), and 140.20 (95% CI, 55.92-351.54), respectively. The area under the HSROC for this test for identifying influenza A was similar to that for identifying influenza B. Age was considered a probable source of heterogeneity. Conclusion: Pooled sensitivities of the Quidel Sofia rapid influenza FIA for identifying influenza A and B did not quite meet the target level (≥80%). Thus, caution is needed when interpreting data of this study due to substantial betweenstudy heterogeneity.

Serum Levels of IL-$1\beta$, IL-2 and IL-6 in Functional Gastrointestinal Disorders in Relation to Depression and Stressful Life Events (기능성 위장관 장애에서 혈청 IL-$1\beta$, IL-2, IL-6의 변화 : 우울증 및 스트레스와의 상관성)

  • Park, Si-Sung;Lee, Sang-Shin;Park, Moo-In;Koo, Ja-Young;Park, Je-Min;Kim, Myung-Jung
    • Korean Journal of Psychosomatic Medicine
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    • v.10 no.2
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    • pp.130-141
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    • 2002
  • Objectives : Stress induces alteration of gastrointestinal function in which interleukin (IL) and hypothalamo-pituitary-axis are involved. Depression again is associated with functional gastrointestinal disorders (FGID) and interleukins. The author attempted to look into the role of interleukins in the FGID also the association of depression and stress in this context. Methods : Entered were 20 patients with FGID, diagnosed by Rome IT criteria and 20 healthy controls. Depression was measured by Hamilton Rating Scale for Depression (HAM-D), and stress of the last one year by Schedule of Recent Experience (SRE). Serum levels of IL-$1\beta$, IL-2, IL-6 were measured by Enzyme-linked Immunosorbent Assay and serum cortisol by Fluorescence Polarization Immunoassay. The results were as follows : Results: 1) Serum levels of IL-$1\beta$, IL-2 were significantly lower in the patients with FGID than those in the controls, but level of IL-6 did not differ between two groups. 2) The patient group showed significantly higher level of serum cortisol as well as higher degrees of depression and stress. 3) Positive correlation was noted between depression and serum cortisol, and between depression and IL-2. A trend of negative correlation was seen between depression IL-$1\beta$. Positive correlation was noted between SRE and IL-6 and IL-6 and serum cortisol. Conclusions : It was concluded that FGID might be related to depression and stress. Changes of the interleukins might be involved with elevated cortisol level.

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Analysis of Methamphetamine and Amphetamine in Oral Fluid of Eleven Drug Abusers (마약남용자 11명의 타액 중 메스암페타민의 분석)

  • Kim, Eun-Mi;Lee, Ju-Seon;Choi, Hye-Young;Choi, Hwa-Kyung;Chung, Hee-Sun
    • YAKHAK HOEJI
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    • v.52 no.6
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    • pp.419-425
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    • 2008
  • A qualitative and quantitative analytical method was developed for detection of methamphetamine (MA) and its main metabolite amphetamine (AM) in oral fluid. Oral fluids of eleven drug abusers were provided by Police, specimens were collected by stimulation with a cotton swab treated with 20 mg of citric acid ($Salivette^{(R)}$; Sarstedt, USA). As the preliminary test, oral fluid samples were screened for amphetamines by Fluorescence Polarization Immunoassay (TDxFLx, Abbott Co.). Extraction for MA was performed using solid-phase extraction (SPE) by $RapidTrace^{TM}$ (Zymark, USA) with mixed mode cation exchange cartridge, CLEAN $SCREEN^{(R)}$ (130 mg/3 ml, UCT) after dilution with phosphate buffer. Samples were evaporated and derivatized by pentafluoropropionic acid anhydride (PFPA). Quantitation of MA and AM was performed by gas chromatography-mass spectrometry (GC-MS) using selective ion monitoring (SIM), the quantitation ions were m/z 204 (MA), 208 (MA-$D_5$), 190 (AM) and 194 (AM-$D_5$). The selectivity, linearity of calibration, limit of detection (LOD) and quantification (LOQ) within- and between day precision, accuracy and recoveries were examined as parts of the method validation. All oral fluid samples gave positive results to immunoassay for MA (cut-off level, 50 ng/ml as d-amphetamine). Concentrations of MA and AM by GC-MS in eleven samples were ranged 104.2${\sim}$4603.3 ng/ml and 32.4${\sim}$268.6 ng/ml, respectively. Extracted calibration curves of MA and AM were linear over the two concentration range of 1${\sim}$100 and 50${\sim}$1000 ng/ml with correlation coefficient of above 0.999. LOQ of MA and AM was 1 and 3 ng/ml, respectively. The intraand inter-day run precisions (CV) for MA and AM were less than 10%, and the accuracies (bias) for MA and AM were also less than 10% at the two different concentrations 5 and 100 ng/ml at low calibration range, 50 and 1000 ng/ml at high calibration range. The absolute recoveries of MA and AM at low and high calibration ranges were more than 82% and 75%, respectively. In this study the qualitative and quantitative analytical method of MA in oral fluid was established. Oral fluid testing may detect drug use in past hours because of its shorter detection window than urine, and be useful in post-accident situations. So oral fluids will be most useful for testing drug abuse in the driving under the influence of drug (DUID) as the alternative specimens of urine.

Quantitative Analysis of Methotrexate using Hitach-7600 P-module (Hitach-7600 P-module을 이용한 Methotrexate 정량분석 평가)

  • Kim, Min-Eui;Cha, Kyong-Ho;Kim, Seung-Hee;Kim, Nam-Joo;Chae, Hyo-Jin
    • Korean Journal of Clinical Laboratory Science
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    • v.41 no.4
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    • pp.167-172
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    • 2009
  • Methotrexate (MTX) in one of the antineoplastic drug and it is known to effective to management of acute lymphoblastic leukemia in children, management of choriocarcinoma and related trophoblastic tumors in women, management of carcinomas of the breast, tongue, pharynx, and tests, maintenance of remission in leukemia and treatment of serve, debilitating psoriasis. Intermediate to high-dose methotrexate administration followed by leucovorin rescue is effective in treatment of carcinoma of the lung and osteogenic sarcoma. Intrathecal administration is effective in treating meningeal leukemia or lymphoma. There are FPIA (Fluorescence polarization immunoassay) and EMIT (Enzyme multiplied immunotechique) methods that measure for MTX. We evaluated the FPIA and EMIT methods. MTX were measured by Hitachi-7600 P-module using EMIT and FPIA using TDX in the sera 60 patients. The performance characteristics evaluated were, light influence, linearity, comparison with FPIA. Also, precision evaluated were three level controls through put following CLSI evaluation protocols (EP10-A). When the MTX value of $4.16{\pm}5.78{\mu}{\mu}mol/L$ (mean, SD) by the Hitachi-7600 P-module was compared with that of $4.05{\pm}5.47{\mu}{\mu}mol/L$ by FPIA, coefficients of correlation of 0.988 was obtained. The regression equation was Y (Hitachi-7600 P-module) = 0.9408 x (FPIA) + 0.1316 (r=0.9885, n=60). CVs of MTX measured by Hitachi 7600 P-module was 6.78% at $0.33{\mu}{\mu}mol/L$, 0.96% at $1.16{\mu}{\mu}mol/L$, and 0.96% at $8.04{\mu}{\mu}mol/L$. The precision was excellent in each group. The linearity was acceptable. We evaluated that MTX is light-sensitive on prolonged exposure to direct sunlight. Comparing with the FPIA using TDX, the Hitachi-7600 P-module using EMIT showed good coefficient of correlation and precision. Therefore the Hitachi-7600 P-module can replace the FPIA for quantitative analysis of MTX.

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