• Title/Summary/Keyword: immunoassay

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A New Concept for Efficient Sensitivity Amplification of a QCM Based Immunosensor for TNF-α by Using Modified Magnetic Particles under Applied Magnetic Field

  • Bahk, Yeon-Kyoung;Kim, Hyung-Hoon;Park, Deog-Su;Chang, Seung-Cheol;Go, Jeung-Sang
    • Bulletin of the Korean Chemical Society
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    • v.32 no.12
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    • pp.4215-4220
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    • 2011
  • This study introduces a new concept for a simple, efficient and cheap sensitivity amplification of a Quartz Crystal Microbalance (QCM) based immunosensor system for the detection of tumor necrosis factor-alpha (TNF-${\alpha}$, TNF) by using an in-built magnetic system. The frequency shift due to the applied magnetic field was successfully observed on magnetic particles labeled detection antibodies, anti-human TNF-${\alpha}$, which were bound to the immunologically captured TNF-${\alpha}$ on the gold coated quartz crystals. In the present system, the magnitude of frequency shift depends on both the strength of magnetic field and the amount of target antigen applied. Significant signal amplification was observed when the additional built-in residual stress generated by the modified magnetic particles under the magnetic field applied. Used in conjunction with a sandwich type non-competitive immunoassay format, the lower detection limit was calculated to be 25 $ngmL^{-1}$ and showed good linearity up to TNF-${\alpha}$ concentrations as high as 2.0 ${\mu}gmL^{-1}$. The sensitivity, most importantly, was improved up to 4.3 times compared with the same QCM system which was used only an antigen-antibody binding without additional magnetic amplification.

A Study of Lupus Anticoagulants and Anticardiolipin Antibodies in Patients with Infertility and Recurrent Spontaneous Abortion (불임환자와 반복자연유산 환자에서 루프스 항응고인자와 항카디오리핀 항체에 대한 연구)

  • Nam, Yoon-Sung;Cha, Kwang-Yul;Baek, Jin-Young;Kim, Nam-Keun;Kang, Myung-Seo;Oh, Do-Yeon
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.29-35
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    • 2002
  • Objective : To report the prevalence of lupus anticoagulants and anticardiolipin antibodies in patients with recurrent spontaneous abortion and infertility. Material and Method: Lupus anticoagulants and anticardiolipin antibodies were analyzed by Diluted Russell's Viper Venom Test (DRVVT) and solid phase enzyme immunoassay, respectively. Results : In 200 patients with infertility, there were 6 cases (3%) with positive lupus anticoagulants or anticardiolipin antibodies. Of these, 3 patients (1.5%) showed positive lupus anticoagulants and anticardiolipin antibodies, respectively. In 120 patients with recurrent spontaneous abortion, there were 13 cases (10.8%) of positive lupus anticoagulants or anticardiolipin antibodies. Of these, one patient (1%) showed lupus anticoagulants and 12 patients (10%) showed anticardiolipin antibodies. But in two groups, there was no cases with positive lupus anticoagulants and anticardiolipin antibodies. Conclusion: Lupus anticoagulants and anticardiolipin antibodies are definite cause of recurrent spontaneous abortion. There has been a speculation that they might be associated with infertility and repeated IVF failures. But it was found that the role of lupus anticoagulants and anticardiolipin antibodies in these cases are not clear.

Analysis on Causal Factors Affecting the Stress of Pilots by the Environmental Differences between Live-Virtual Simulation (Live-Virtual 시뮬레이션 환경차이에 따른 조종사 스트레스 유발요인 분석)

  • Kim, Jinju;Kim, Sungho;Seol, Hyeonju;Jee, Cheolkyu;Hong, Youngseok
    • Journal of Korean Society of Industrial and Systems Engineering
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    • v.39 no.1
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    • pp.31-36
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    • 2016
  • Recently, Live-Virtual-Constructive (L-V-C) integrate training system has proposed as a solution for the problems such as limitation of training areas, increase of mission complexity, rise in oil prices. In order to integrate each training system into the one effectively, we should solve the issue about stress of pilots by the environmental differences between Live and Virtual simulation which could be occurred when each system is connected together. Although it was already examined in previous study that the psychological effects on pilots was occurred by the environmental differences between actual and simulated flights, the study did not include what the causal factors affecting psychological effects are. The aim of this study is to examine which environmental factors that cause pilots' psychological effects. This study analyzed the biochemical stress hormone, cortisol to measure the pilots' psychological effects and cortisol was measured using Enzyme-linked immunoassay (EIA). A total of 40 pilots participated in the experiment to compare the differences in pilots' cortisol response among live simulation, virtual simulation, and the virtual simulation applying three environmental factors (gravity force, noise, and equipment) respectively. As a result, there were significant differences in cortisol level when applied the gravity force and equipment factors to the virtual simulation, while there was no significant difference in the case of the noise factor. The results from this study can be used as a basis for the future research on how to make L-V system by providing minimum linkage errors and design the virtual simulator that can reduce the differences in the pilots' psychological effects.

An Experimental Study on the Anti-inflammatory Effects of Cheonggisan Extract in Allergic Late Inflammation (알레르기 후기 반응 염증 억제효과에 관한 청기산(淸肌散)의 실험적 연구)

  • Jo, Suk-Yong;Kang, Min-Seo;Kim, Yong-Min;Kim, Hee-Taek
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.3
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    • pp.95-105
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    • 2016
  • Objectives : Allergic diseases have a various symptoms of hyperresponsiveness and recently hyperresponsive reaction in the chronic phase is reported as the important mechanisms. Cheonggisan(CGS) is used in oriental clinics for curing various skin diseases due to effect of controlling of pruritus. There have been studies on the anti-allergic effect and anti-inflammatory effect of CGS, but there had no study of anti-allergic effects in allergic late inflammation of CGS, so we aimed to find out the effects of CGS in allergic late inflammation in our study.Methods : To investigate the anti-allergy effect and anti-inflammatory effect of CGS, RAW 264.7 macrophage cells and CSG water-extracts were used. Cytotoxic effect of CSG was examined by MTT assay, an oxidative product of NO was measured in the culture medium by the Griess reagent assay. The level of prostaglandin E2(PGE2) was measured by competitive enzyme-linked immunoassay. Cytokine(PGE2, IL-1β, IL-6, TNF-α) was measured by Bio-Plex suspension assay system and quantitative multiplexed cytokine/chemokine assay.Results : We investigated that there was no cytotoxic effect of CGS water-extract at any levels of concentration on RAW 264.7 macrophage cells by MTT assay. CGS water-extracts significantly suppressed the levels of the inflammatory mediators such as NO and PGE2, cytokine of IL-1β, TNF-α at the level of 400 ㎍/㎖ CGS concentration. But there was no significant effect on IL-6 production suppression.Conclusions : These results suggest that CSG water-extract has and anti-inflammatory effects in allergic reaction. These properties may contribute to the allergic diseases and inflammatory related disease care.

Effects of Auklandia Lappa on Dextran Sulfate Sodium-Induced Murine Colitis (당목향(唐木香)이 DSS(Dextran sulfate sodium)로 유발된 염증성 장질환 동물모델에 미치는 영향)

  • Kim, So-Yeon;Park, Jae-Woo;Ryu, Bong-Ha
    • The Journal of Internal Korean Medicine
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    • v.34 no.2
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    • pp.134-146
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    • 2013
  • Objectives : Auklandia Lappa (ALE) is one of the herbs used frequently to treat abdominal pain and diarrhea and reported anti-inflammatory activity by suppressing proinflammatory cytokines. This study was designed to investigate whether ALE could show protective activities on experimental colitis induced by dextran sulfate sodium (DSS) models. Methods : Colitis was induced by DSS in Balb/c mice. ALE 10, 30, 100 and 300 mg/kg were orally administered twice a day for 7 days in DSS model. Mice weight was measured daily. Scoring of clinical findings was measured every other day. Colon length, edema, fecal blood and histological damages were assessed at day 7 in DSS model. In histological analysis, we checked cryptal glands, surface epithelium, submucosa, transmural, stroma and scored degree of inflammatory cell damage by modified histological scoring. We also calculated cytokines concentrations including IFN-${\gamma}$, TNF-${\alpha}$, IL-6, IL-$1{\beta}$, IL-17, IL-23, IL-10 and TGF-${\beta}1$ by Biometric Multiplex Cytokine Profiling method. Results : ALE showed the protective effects on DSS-induced experimental colitis. ALE inhibited shortening of colon length and histological damages of colon does-dependently, but it did not inhibit weight loss. ALE also inhibited IFN-${\gamma}$ and IL-6 expression, and upregulated cytokines (IL-10, TGF-${\beta}1$) related to regulatory T cell differentiation and proliferation. Conclusions : The current results demonstrate the clinical utility of ALE in traditional medicine and indicate the possibility of potent drug development of inflammatory bowel diseases from natural products. Further investigations for exact mechanisms will be needed.

Dot Blot Assay for Screening of Anti-hantavirus Antibodies by Using Nucleocapsid Protein of Hantaan Virus (한탄바이러스 핵단백질을 이용한 항 한타바이러스 항체 검색용 Dot Blot Assay)

  • Cho, Hae-Wol;Chung, Yeun-Jun;Kim, Chung-Lim;Ban, Sang-Ja;Nam, Jae-Hwan;Lee, Hyeong-Woo;Lee, Yoo-Jin;Kim, Eun-Jung
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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    • pp.59-65
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    • 1996
  • For easy and rapid screening of hemorrhagic fever with renal syndrome (HFRS) without any laboratory equipment, dot blot enzyme immunoassay was developed and tried to detect anti-hantavirus antibodies. The nucleocapsid protein of Hantaan virus was isolated by affinity chromatography and used for making the dot strip. 28 of 29 Hantaan virus infected sera showed positive signals and 21 of 22 HFRS negative sera showed no positive signals. Anti-Seoul virus monoclonal antibody also exibited positive signal but the intensity of colorization was approximately 5 fold less than that of anti-Hantaan monoclonal antibody. The sensitivity of dot blot assay was equal or superior to indirect immunofluorescent assay (IFA) or ELISA test. Overall, the screening results with dot blot assay showed 92.2 % of concordance with IFA or ELISA test. This results suggests that dot blot assay could be applied a tool for easy and rapid screening of HFRS.

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Pharmacokinetics and Tissue Distribution of Recombinant Human Erythropoietin (DA-3285) in the Laboratory Animals (Recombinant human erythropoietin (DA-3285)의 실험동물에서의 약동력학 및 조직분포)

  • 심현주;이응두;이종진;김흥재;이상득;이성희;김원배;양중익
    • Biomolecules & Therapeutics
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    • v.4 no.1
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    • pp.78-83
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    • 1996
  • The pharmacokinetics and tissue distribution of DA-3285 (recombinant human erythropoietin, recently manufactured by Research Laboratories of Dong-A Pharmaceutical Company) were studied in the laboratory animals. The plasma, urine, and tissue concentration of DA-3285 were measured by a double-antibody sandwich enzyme immunoassay. After intravenous administration of DA-3285, 20, 100, 500 and 2500 units/kg to rats, the plasma concentrations declined polyexponentially with the terminal half-lives of 2.15, 2.10, 2.31, and 2.35 hr, respectively. Total body clearance (20.7∼26.6 mι/hr/kg) and apparent volume of distribution at steady state (57.2∼70.1 mι/kg) were independent of the dose and AUC increased proportionally with the dose. The renal clearance was much lower than total body clearance, suggesting that extrarenal clearance, presumably metabolism , plays a significant role in elimination of DA-3285. In all rat tissues, the tissue to plasma ratios were smaller than unity, indicating less affinity of DA-3285 to rat tissues and was proved by considerably less value of Vdss. After 3 times a week for consecutive 3 weeks i.v. administration of DA-3285, 100 units/kg to rats, the plasma concentrations and pharmacokinetic parameters of DA-3285 were not significantly different from those in a single administration. After s.c. administration to the rat, plasma concentrations of DA-3285 peaked at 6 hr and the extent of bioavailability was 26.7%. In mice, rabbits and dogs, at DA-3285 dose of 100 units/kg, the mean terminal haw-lives were 2.78, 3.05, and 4.01 hr, respectively. Compared with reported data in the literatures, DA-3285 has similar properties to rh-EPO manufactured by other companies in view of pharmacokinetics.

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The Relation between Exposure to Environmental Tobacco Smoke and the Quantity of Cotinine in the Urine of School Children in Taif City, Saudi Arabia

  • Desouky, Dalia El Sayed;Elnemr, Gamal;Alnawawy, Ali;Taha, Azza Ali
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.1
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    • pp.139-145
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    • 2016
  • Environmental tobacco smoke (ETS) is a major public health problem for all ages. Despite the high prevalence of smoking among the Saudi population, there is limited information about levels of urinary cotinine in Saudi children exposed to SHS. The aim of the study was to assess the exposure of schoolchildren to ETS, and measure their urinary cotinine levels. Multistage cluster sampling was carried out, where schoolchildren from 4 schools were randomly chosen from primary schools in Taif city. A questionnaire including questions on SHS exposure and smoking rules in the residence were sent to students parents/guardians. Urine samples were taken and analyzed for total cotinine using chemiluminescent immunoassay. Of the studied children, 38.4% had a smoking father, 61.8%, 41.2% and 49.3% of them were exposed to ETS indoors, outdoors and both indoors and outdoors respectively. The mean urinary cotinine was significantly higher among children exposed to ETS compared to unexposed children. Urinary cotinine levels in children with both indoor and outdoor exposure was significantly higher compared with its level in children with single exposure. A significant positive correlation was found between urinary cotinine concentrations and the number of cigarette packs smoked by parents, and the number of smokers in the residence. The mean urinary cotinine level was significantly higher in children who reported no smoking rules at the residence.. The study revealed a high exposure of Saudi children to ETS. An antismoking media awareness campaign on the harmful effects of ETS should be carried out, in addition to family counseling programs targeted to parents to protect their children from ETS.

Comparative Serum Proteomic Analysis of Serum Diagnosis Proteins of Colorectal Cancer Based on Magnetic Bead Separation and MALDI-TOF Mass Spectrometry

  • Deng, Bao-Guo;Yao, Jin-Hua;Liu, Qing-Yin;Feng, Xian-Jun;Liu, Dong;Zhao, Li;Tu, Bin;Yang, Fan
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.10
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    • pp.6069-6075
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    • 2013
  • Background: At present, the diagnosis of colorectal cancer (CRC) requires a colorectal biopsy which is an invasive procedure. We undertook this pilot study to develop an alternative method and potential new biomarkers for diagnosis, and validated a set of well-integrated tools called ClinProt to investigate the serum peptidome in CRC patients. Methods: Fasting blood samples from 67 patients diagnosed with CRC by histological diagnosis, 55 patients diagnosed with colorectal adenoma by biopsy, and 65 healthy volunteers were collected. Division was into a model construction group and an external validation group randomly. The present work focused on serum proteomic analysis of model construction group by ClinProt Kit combined with mass spectrometry. This approach allowed construction of a peptide pattern able to differentiate the studied populations. An external validation group was used to verify the diagnostic capability of the peptidome pattern blindly. An immunoassay method was used to determine serum CEA of CRC and controls. Results: The results showed 59 differential peptide peaks in CRC, colorectal adenoma and health volunteers. A genetic algorithm was used to set up the classification models. Four of the identified peaks at m/z 797, 810, 4078 and 5343 were used to construct peptidome patterns, achieving an accuracy of 100% (> CEA, P<0.05). Furthermore, the peptidome patterns could differentiate the validation group with high accuracy close to 100%. Conclusions: Our results showed that proteomic analysis of serum with MALDI-TOF MS is a fast and reproducible approach, which may provide a novel approach to screening for CRC.

Research on the Relationship Between Serum Levels of Inflammatory Cytokines and Non-small Cell Lung Cancer

  • Song, Xiao-Yun;Zhou, Shi-Jie;Xiao, Ning;Li, Yun-Song;Zhen, De-Zhi;Su, Chong-Yu;Liu, Zhi-Dong
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.8
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    • pp.4765-4768
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    • 2013
  • Aims: This study was conducted to evaluate the levels of TNF-${\alpha}$, IL-6, IL-8 and VEGF in serum of patients with non- small cell lung cancer, for assessing their possible diagnostic and prognostic roles. Methods: We enrolled 48 patients newly diagnosed with non-small cell lung cancer and 40 healthy controls. TNF- ${\alpha}$, IL-6 and IL-8 levels were measured in the serum of all the subjects with specific radioimmunoassay kits, while EGF was analyzed by sandwich enzyme immunoassay techniques. Results: A statistically significant difference was observed between lung cancer patients and the control group regarding the values of TNF-${\alpha}$, IL-6, IL-8 and VEGF in serum. Moreover, TNF-${\alpha}$, IL-8 and VEGF levels were higher in patients with advanced stages compared to early stages. In addition, higher serum levels of TNF-${\alpha}$, IL-6, IL-8 and VEGF were found in smokers than in non-smokers, both in patients and controls. Conclusion: Serum levels of TNF-${\alpha}$, IL-6, IL-8 and VEGF were all elevated in lung cancer patients, suggesting that inflammatory cytokines could be jointly used as a screening tool. Though TNF-${\alpha}$, IL-8 and VEGF levels were related to advanced disease, long-term survival studies of NSCLC patients should be performed to confirm whether they can act as biomarkers of advanced disease. In addition, smoking would be an important contributor to the processes of inflammation and lung cancer.