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Sample records for luminol

  1. Application of forensic luminol for blood detection in endodontic files

    Directory of Open Access Journals (Sweden)

    Rodrigo ARRUDA-VASCONCELOS

    Full Text Available Abstract Introduction Blood is a biological material with high potential of infectious transmission in dental environments, including herpes simplex, hepatitis and AIDS. Aim To investigate the efficacy of luminol in detecting blood in endodontic files before and after the sterilization process. Material and method Luminol was used to investigate the presence or absence of traces of blood tissue in 50 endodontic files, visible to naked eye or not, after performing endodontic treatment and after the cleaning/sterilization process. The results obtained were tabulated and statistically analyzed by using the Friedman’s test at a significance level of 5% (p<0.05. Result By naked eye, it was found that 31/50 files showed no trace of blood, 8/50 showed a slight presence of blood and 11/50 showed a considerable presence of blood after endodontic treatment. After the use of luminol, however, 16/50 endodontic files showed no trace of blood, 19/50 showed a slight presence of blood and 15/50 showed a considerable presence of blood. After the cleaning and sterilization process, no blood was detected in the files. Conclusion It was concluded that the luminol solution is effective in detecting blood tissue in endodontic files as well as in validating the cleaning/sterilization process.

  2. Luminol, horseradish peroxidase, and glucose oxidase ternary functionalized graphene oxide for ultrasensitive glucose sensing.

    Science.gov (United States)

    Li, Fang; Ma, Wenjing; Liu, Jiachang; Wu, Xiang; Wang, Yan; He, Jianbo

    2018-01-01

    Luminol, horseradish peroxidase (HRP), and glucose oxidase (GOx) ternary functionalized graphene oxide (HRP/GOx-luminol-GO) with excellent chemiluminescence (CL) activity and specific enzymatic property was prepared via a simple and general strategy for the first time. In this approach, luminol functionalized GO (luminol-GO) was prepared by gently stirring GO with luminol. Then HRP and GOx were further co-immobilized onto the surface of luminol-GO by storing HRP and GOx with luminol-GO at 4 °C overnight, to form HRP/GOx-luminol-GO bionanocomposites. The synthesized HRP/GOx-luminol-GO could react with H 2 O 2 generated from GOx catalyzed glucose oxidization reaction, to produce strong CL emission in the presence of co-immobilized HRP. Thus, we developed an ultrasensitive, homogeneous, reagentless, selective, and simple CL sensing system for glucose detection. The resulting biosensors exhibited ultra-wide linear range from 5.0 nM to 5.0 mM, and an ultra-low detection limit of 1.2 nM, which was more than 3 orders of magnitude lower than previously reported methods. Furthermore, the sensing system was successfully applied for the detection of glucose in human blood samples.

  3. Electrogenerated chemiluminescence of luminol using low-cost electrodes

    International Nuclear Information System (INIS)

    Salgado, Guillermo; Navarrete, Jose; Bustos, Carlos; Sanchez, Cristian; Ugarte, Ricardo

    2006-01-01

    The purpose of the study was to observe eletrogenerated chemiluminescence (ECL) of luminol using different materials as electrodes such as wires, nails, coins, razor blades, etc. Based on the experimental observations and the mechanisms proposed in the literature, students should be capable of analyzing their results in order to understand the phenomena studied. Students could then elaborate a mechanism of action consistent with the experimental results which could rationalize the formation of intermediates in the reaction and the dependence of ECL on solution pH in addition to other points of interest. (author)

  4. Poly(luminol) based sensor array for determination of dissolved chlorine in water

    Czech Academy of Sciences Publication Activity Database

    Szili, M.; Kašík, Ivan; Matějec, Vlastimil; Nagy, G.; Kovács, B.

    -, č. 192 (2014), s. 92-98 ISSN 0925-4005 Institutional support: RVO:67985882 Keywords : Chemiluminescence * Electropolymerization * Luminol Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 4.097, year: 2014

  5. Quantitation of immunoadsorbed flavoprotein oxidases by luminol-mediated chemiluminescence.

    Science.gov (United States)

    Hinkkanen, A; Maly, F E; Decker, K

    1983-04-01

    The detection of the flavoenzymes 6-hydroxy-L-nicotine oxidase and 6-hydroxy-D-nicotine oxidase at the sub-femtomol level was achieved by coupling the reaction of the immunoadsorbed proteins to the peroxidase-catalysed oxidation of luminol. The H2O2-producing oxidases retained their full activity when bound to the respective immobilized antibodies. This fact allowed the concentration of the enzymes from very dilute solutions and the quantitative assay of their activities in the microU range. Due to strict stereoselectivity and the absence of immunological cross-reactivity, the two flavoproteins could be determined in the same solution. This method was used to measure the 6-hydroxy-D-nicotine oxidase and 6-hydroxy-L-nicotine oxidase activities in Escherichia coli RR1 and different Arthrobacter strains cultured under non-inducing conditions. The same activity ratio of 6-hydroxy-L-nicotine oxidase/6-hydroxy-D-nicotine oxidase as in D L-nicotine-induced cells of A. oxidans was observed in non-induced wild type and in riboflavin-requiring (rf-) mutant cells of this aerob.

  6. The chemiluminescence of luminol in aqueous solutions, a pulse radiolytic study

    International Nuclear Information System (INIS)

    Lind, J.

    1980-01-01

    The mechanism of chemiluminescence was studied when the luminol radical and superoxide were generated simultaneously. Hydroperoxide was formed at the first reaction step with a pKa value between 11 and 12. The dissociated form was found to undergo a monomolecular reaction leading to light generation and having a rate constant > 2 x 10 5 s -1 . The protonated form had the rate constant of approx. 2000 s -1 and formed a product which absorbed around 390 nm without light emission. The reaction of the two-electron oxidation product of luminol, 5-aminophthalazine-1, 4 dione (azaquinone), with hydrogenperoxide was stoichiometrically equivalent to the recombination of superoxide with the luminol radical. The pulseradiolytic generation of the chlorinedioxide radical ClO 2 in aqueous solution of sodium chlorite is described, and the oxidation of luminol by ClO 2 in two steps is reported: 1. Radical adducts are formed. 2. The reaction of ClO 2 x radical with the adduct to form azaquinone. The chemiluminescent reaction between the azaquinone and hydrogenperoxide consists of a second order reaction between HO 2 and the azaquinone to form a hydroperoxide followed by a monomolecular rearrangement of the dissociated form of the latter. Its acidbase equilibrium has a pKa of 10.6. A nitrogencentered structure is ascribed to the hydroperoxide formed by cross-recombination of the luminolradical and superoxide while the reaction between HO 2 and azaquinone yields a carboncentered hydroperoxide. The existence of two different emitters is evidenced by the following observations. 1. A slight difference in chemiluminescence spectra. 2. The formation of an endproduct with pKa approx. 10.2 in the luminolradical superoxide system. 3. The formation of an endproduct which acts as an efficient radial scavenger in the luminol radical superoxide system. (G.B.)

  7. Forensic Luminol Blood Test for Preventing Cross-contamination in Dentistry: An Evaluation of a Dental School Clinic

    Directory of Open Access Journals (Sweden)

    Marcelo Carlos Bortoluzzi

    2014-01-01

    Conclusions: We suggest that the luminol method is suitable for identifying contamination with invisible blood traces and this method may be a useful tool to prevent cross-contamination in the dental care setting.

  8. A quantitative method for determining a representative detection limit of the forensic luminol test for latent bloodstains.

    Science.gov (United States)

    Cassidy, Brianna M; Lu, Zhenyu; Martin, Jennifer P; Tazik, Shawna K; Kellogg, Katie W; DeJong, Stephanie A; Belliveau, Elle O; Kilgore, Katherine E; Ervin, Samantha M; Meece-Rayle, Mackenzie; Abraham, Alyssa M; Myrick, Michael L; Morgan, Stephen L

    2017-09-01

    The luminol test has been used for over 60 years by forensic investigators for presumptive identification of blood and visualization of blood splatter patterns. Multiple studies have estimated the limit of detection (LD) for bloodstains when luminol is employed, with results ranging from 100× to 5,000,000× dilute. However, these studies typically have not identified and controlled important experimental variables which may affect the luminol LD for bloodstains. Without control of experimental parameters in the laboratory, variables which affect the potential of presumptive bloodstain test methods remain largely unknown, and comparisons required to establish new, more powerful detection methods are simply impossible. We have developed a quantitative method to determine the relationship between the amount of blood present and its reaction with luminol by measuring, under controlled conditions, the resulting chemiluminescent intensity with a video camera, combined with processing of the digital intensity data. The method resulted in an estimated LD for bloodstains on cotton fabric at ∼200,000× diluted blood with a specific luminol formulation. Although luminol is the focus of this study, the experimental protocol used could be modified to study effects of variables using other blood detection reagents. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. A novel chemiluminescence assay of organophosphorous pesticide quinalphos residue in vegetable with luminol detection

    Directory of Open Access Journals (Sweden)

    Yao Xin

    2010-06-01

    Full Text Available Abstract Background Organophosphorous pesticides are the most popular pesticides used in agriculture. As acetylcholinesterase inhibitors, organophosphorous pesticides are toxic organic chemicals. The control and detection of organophosphorous pesticide residue in food, water, and environment therefore plays a very important role in maintaining physical health. A sensitive, rapid, simple chemiluminescence(CL method has been developed for the determination of quinalphos based on the reaction of quinalphos with luminol-H2O2 in an alkaline medium. The method has been applied to detection of quinalphos in vegetable samples with satisfactory results. Results The CL method for the determination of organophosphorous pesticide quinalphos is based on the phenomenon that quinalphos can apparently enhance the CL intensity of the luminol-H2O2 system. The optimal conditions were: luminol concentration 5.0 × 10-4 mol/L, H2O2 concentration 0.05 mol/L.pH value 13. In order to restrain the interference from metal ions, 1.0 × 10-3 mol/L of EDTA was added to the luminol solution. The possible mechanism was proposed. Conclusion Under the optimum reaction conditions, CL was linear with the concentration of quinalphos in the range of 0.02 μg/mL -1.0 μg/mL and the detection limit was 0.0055 μg/mL (3σ. This method has been successfully applied to the detection of quinalphos in vegetable samples. According to the experimental data, the average recoveries for quinalphos in cherry tomato and green pepper 97.20% and 90.13%. Meanwhile, the possible mechanism was proposed.

  10. Application of Box-Behnken design in the optimization of catalytic behavior of a new mixed chelate of copper (II) complex in chemiluminescence reaction of luminol

    International Nuclear Information System (INIS)

    Khajvand, Tahereh; Chaichi, Mohammad Javad; Nazari, OmLeila; Golchoubian, Hamid

    2011-01-01

    In this work, we observed an enhancement of chemiluminescence (CL) emission of luminol when a new mixed chelate of copper complex (N-(2-(2-aminoethylamino)ethyl)-1H-pyrrole-2-carboxamide-Cu(II)) was mixed with a solution containing luminol in methanol/water. The Box-Behnken design matrix and response surface methodology (RSM) have been applied to design the experiments to evaluate the interactive effects of the three most important operating variables-luminol (10 -4 -10 -2 M), fluorescein (10 -5 -10 -3 M) and hydrogen peroxide (1-3 M) concentrations on the CL emission of luminol. The total 15 experiments were conducted in the present study towards the construction of a quadratic model. Independent variables luminol and hydrogen peroxide have significant value P F less than 0.0500 indicate that model terms are significant for the CL emission of luminol. The regression equation coefficients were calculated and the data fitted to a second-order polynomial equation for CL emission of luminol. The new introduced inorganic catalyst of luminol CL reaction can be effect more than that of the common ones such as potassium hexacyanoferrate (III) and copper (II) acetate. - Research highlights: → In this study we introduce a new mixed chelate of copper complex as a catalyst of luminol chemiluminescence (CL) reaction. → The copper complex (N-(2-(2-aminoethylamino)ethyl)-1H-pyrrole-2-carboxamide-Cu(II)) catalyst luminol reaction more than that of copper acetate and potassium hexacyanoferrate (III). → The Box-Behnken design matrix and response surface methodology are used for prediction of CL intensity of luminol. → There are good correlation between experimental and expected CL intensity that predicted by the theoretical model. → Fluorescein used as a fluorescer in the luminol CL reaction in presence of the new catalyst.

  11. Determination of sibutramine with a new sensor based on luminol electrochemiluminescence

    International Nuclear Information System (INIS)

    Wang Shaowei; Yu Jinghua; Wan Fuwei; Ge Shenguang; Yan Mei; Zhang Meng

    2011-01-01

    A flow injection electrochemiluminescence (FI-ECL) analysis method for the determination of sibutramine in the presence of luminol was studied under conventional cyclic voltammetry in alkaline Na 2 CO 3 -NaHCO 3 buffer solution (pH 8.0-12.0). This method is based on the enhanced ECL of luminol-sibutramine. Meanwhile, in order to overcome the drawbacks of conventional cells, a FI cell was designed, which is reusable and has a great improvement in sensitivity and selectivity for ECL analysis. Under the optimal experimental conditions, the enhanced ECL intensity was linearly related to the concentration of sibutramine in the range 1.0x10 -8 -1.0x10 -6 g mL -1 with a detection limit of 2.48x10 -9 g mL -1 and a correlation coefficient (R) of 0.9995. The relative standard deviation (RSD) for 1.0x10 -7 g mL -1 samples was 2.1% (n=11). The possible mechanism discussed. The proposed FI-ECL method has been successfully applied to the determination of sibutramine in diet pill samples. - Highlights: → An electrochemiluminescence (ECL) system was studied to determine sibutramine. → A flow injection cell was made to overcome the drawbacks of the conventional ones. → The system was based on the enhanced ECL of luminol-sibutramine. → The system can be satisfactorily applied to determine sibutramine in diet pills.

  12. Self-Catalyzing Chemiluminescence of Luminol-Diazonium Ion and Its Application for Catalyst-Free Hydrogen Peroxide Detection and Rat Arthritis Imaging.

    Science.gov (United States)

    Zhao, Chunxin; Cui, Hongbo; Duan, Jing; Zhang, Shenghai; Lv, Jiagen

    2018-02-06

    We report the unique self-catalyzing chemiluminescence (CL) of luminol-diazonium ion (N 2 + -luminol) and its analytical potential. Visual CL emission was initially observed when N 2 + -luminol was subjected to alkaline aqueous H 2 O 2 without the aid of any catalysts. Further experimental investigations found peroxidase-like activity of N 2 + -luminol on the cleavage of H 2 O 2 into OH • radical. Together with other experimental evidence, the CL mechanism is suggested as the activation of N 2 + -luminol and its dediazotization product 3-hydroxyl luminol by OH • radical into corresponding intermediate radicals, and then further oxidation to excited-state 3-N 2 + -phthalic acid and 3-hydroxyphthalic acid, which finally produce 415 nm CL. The self-catalyzing CL of N 2 + -luminol provides us an opportunity to achieve the attractive catalyst-free CL detection of H 2 O 2 . Experiments demonstrated the 10 -8 M level detection sensitivity to H 2 O 2 as well as to glucose or uric acid if presubjected to glucose oxidase or uricase. With the exampled determination of serum glucose and uric acid, N 2 + -luminol shows its analytical potential for other analytes linking the production or consumption of H 2 O 2 . Under physiological condition, N 2 + -luminol exhibits highly selective and sensitive CL toward 1 O 2 among the common reactive oxygen species. This capacity supports the significant application of N 2 + -luminol for detecting 1 O 2 in live animals. By imaging the arthritis in LEW rats, N 2 + -luminol CL is demonstrated as a potential tool for mapping the inflammation-relevant biological events in a live body.

  13. Formation of Hydrogen Peroxide by Electrochemical Reduction of Molecular Oxygen using Luminol Chemiluminescence

    International Nuclear Information System (INIS)

    Rana, Sohail

    2005-01-01

    Formation of hydrogen peroxide by electrochemical reduction of molecular oxygen was examined by measuring luminol chemiluminescence and absorption spectrum using flow-injection method. Ferryl porphyrin is widely accepted as responsible species to stimulate the emission in hydrogen peroxide/ iron porphyrin/ luminol system. Emission was observed under cathodic potentials (0.05V at pH2.0 and -0.3V at pH11.0) by the electrochemical reduction of aerated electrolytes solution but emission was observed at anodic potentials. Iron porphyrin solution was added at down stream of the working electrode and was essential for the emission. Removal of the dissolved molecular oxygen resulted in the decrease of emission intensity by more than 70%. In order to examine the life time of reduced active species, delay tubes were introduced between working electrode Fe TMPyP inlet. Experimental results suggested the active species were stable for quite a long period. The emission was quenched considerably (>90%) when hydroperoxy was added at the down stream of working electrode whereas the Superoxide dismutase (SOD) had little effect and mannitol had no effect. The spectra at reduction potential under aerated condition were shifted to the longer wavelength (>430nm) compared to the original spectrum of Fe TMPyP (422nm), indicating that the ferryl species were mixed to some extent. These observations lead to the conclusion that hydrogen peroxide was produced first by electrochemical reduction of molecular oxygen which then converted Fe TMPyP into O=FeTMPyP to activate luminol. Comparing emission intensities with the reference experiments, the current efficiencies for the formation of hydrogen peroxide were estimated as about 30-65% in all over the pH range used. (author)

  14. A competitive immunoassay for sensitive detection of small molecules chloramphenicol based on luminol functionalized silver nanoprobe.

    Science.gov (United States)

    Yu, Xiuxia; He, Yi; Jiang, Jie; Cui, Hua

    2014-02-17

    Chloramphenicol (CHL) as a broad-spectrum antibiotic has a broad action spectrum against Gram-positive and Gram-negative bacteria, as well as anaerobes. The use of CHL is strictly restricted in poultry because of its toxic effect. However, CHL is still illegally used in animal farming because of its accessibility and low cost. Therefore, sensitive methods are highly desired for the determination of CHL in foodstuffs. The immunoassays based on labeling as an important tool have been reported for the detection of CHL residues in food-producing animals. However, most of the labeling procedures require multi-step reactions and purifications and thus they are complicated and time-consuming. Recently, in our previous work, luminol functionalized silver nanoparticles have been successfully synthesized, which exhibits higher CL efficiency than luminol functionalized gold nanoparticles. In this work, the new luminol functionalized silver nanoparticles have been used for the labeling of small molecules CHL for the first time and a competitive chemiluminescent immunoassay has been developed for the detection of CHL. Owing to the amplification of silver nanoparticles, high sensitivity for CHL could be achieved with a low detection limit of 7.6×10(-9) g mL(-1) and a wide linear dynamic range of 1.0×10(-8)-1.0×10(-6) g mL(-1). This method has also been successfully applied to determine CHL in milk and honey samples with a good recoveries (92% and 102%, 99% and 107% respectively), indicating that the method is feasible for the determination of CHL in real milk and honey samples. The labeling procedure is simple, convenient and fast, superior to previously reported labeling procedures. The immunoassay is also simple, fast, sensitive and selective. It is of application potential for the determination of CHL in foodstuffs. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Determination of sibutramine with a new sensor based on luminol electrochemiluminescence

    Energy Technology Data Exchange (ETDEWEB)

    Wang Shaowei [Shandong Provincial Key Laboratory of Fluorine Chemistry and Chemical Materials, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Yu Jinghua, E-mail: wangshaowei119@hotmail.co [Shandong Provincial Key Laboratory of Fluorine Chemistry and Chemical Materials, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China); Wan Fuwei; Ge Shenguang; Yan Mei; Zhang Meng [Shandong Provincial Key Laboratory of Fluorine Chemistry and Chemical Materials, School of Chemistry and Chemical Engineering, University of Jinan, Jinan 250022 (China)

    2011-07-15

    A flow injection electrochemiluminescence (FI-ECL) analysis method for the determination of sibutramine in the presence of luminol was studied under conventional cyclic voltammetry in alkaline Na{sub 2}CO{sub 3}-NaHCO{sub 3} buffer solution (pH 8.0-12.0). This method is based on the enhanced ECL of luminol-sibutramine. Meanwhile, in order to overcome the drawbacks of conventional cells, a FI cell was designed, which is reusable and has a great improvement in sensitivity and selectivity for ECL analysis. Under the optimal experimental conditions, the enhanced ECL intensity was linearly related to the concentration of sibutramine in the range 1.0x10{sup -8}-1.0x10{sup -6} g mL{sup -1} with a detection limit of 2.48x10{sup -9} g mL{sup -1} and a correlation coefficient (R) of 0.9995. The relative standard deviation (RSD) for 1.0x10{sup -7} g mL{sup -1} samples was 2.1% (n=11). The possible mechanism discussed. The proposed FI-ECL method has been successfully applied to the determination of sibutramine in diet pill samples. - Highlights: {yields} An electrochemiluminescence (ECL) system was studied to determine sibutramine. {yields} A flow injection cell was made to overcome the drawbacks of the conventional ones. {yields} The system was based on the enhanced ECL of luminol-sibutramine. {yields} The system can be satisfactorily applied to determine sibutramine in diet pills.

  16. Assay of picogram level isocarbophos residue on tangerines and oranges with luminol-albumin chemiluminescence system.

    Science.gov (United States)

    Chen, Donghua; Song, Zhenghua; Lv, Hairu

    2012-12-15

    A sensitive flow injection-chemiluminescence (FI-CL) method for the determination of isocarbophos (ICP) residue on tangerines and oranges was proposed. It was found that the CL intensity from luminol-albumin CL reaction could be obviously quenched in the presence of ICP and the decrease in CL intensity was proportional to the logarithm of ICP concentrations ranging from 1.0 to 1000 pmol L(-1), giving the limit of detection of 0.3 pmol L(-1) (3σ). The proposed procedure was successfully applied to the determination of ICP residue on tangerines and oranges with recoveries varying from 92.0 to 111.0% and RSDs less than 5.0%. The possible CL mechanism of luminol-albumin-ICP reaction was discussed, and ICP to albumin's binding constant (K(D)=1.00 × 10(6) L mol(-1)) and the number of binding sites (n=1.00) were given by the homemade FI-CL model. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. A new luminol chemiluminescence sensor for glucose based on pH-dependent graphene oxide.

    Science.gov (United States)

    Hao, Minjia; Liu, Na; Ma, Zhanfang

    2013-08-07

    In this study, graphene oxide (GO) was found to catalyze the luminol-O2 reaction, which yielded a novel chemiluminescence (CL). Remarkably, the CL emission could be tuned by modulating the pH of the GO dispersion. Transmission electron microscopy, CL spectra, electron spin resonance spectra studies were carried out to investigate the CL mechanism. The results indicate that the CL emission was attributed to the intrinsic catalytic effect of GO acting as the radical generation proliferators and electron transfer accelerators. Based on the GO catalyzed luminol-O2 system, we successfully developed a new CL sensor to detect glucose. Under the optimized conditions, glucose could be assayed in the range of 0.05 mM to 5 mM with a detection limit of 0.044 mM. For the detection of clinical serum samples, it is well consistent with the data determined by commercially available method in hospital, indicating that the new CL method provides a possible application for the detection of glucose in clinical diagnostics.

  18. Enhanced luminol electrochemiluminescence triggered by an electrode functionalized with dendrimers modified with titanate nanotubes

    International Nuclear Information System (INIS)

    Lin, Yanyu; Yang, Yusheng; Dai, Hong; Xu, Guifang; Yang, Ting; Tong, Yuejin; Yang, Caiping; Chen, Guonan

    2013-01-01

    We have constructed a novel electro chemiluminescence (ECL) platform by functionalizing a poly(amidoamine) dendrimer (PAAD) with titanate nanotubes (TiNTs). The PAAD has an open spherical structure that possesses a high density of active groups and thus favors mass transport, while the TiNTs possess excellent electronic conductivity and thus can promote electron transfer on the surface of a glassy carbon electrode (GCE). A study on the intensity and stability of the ECL of luminol on the modified GCE revealed a substantial improvement compared to that of a bare GCE. The effects of the concentration of TiNTs, the pH value of the solution, and of electrochemical parameters on the intensity of the ECL of luminol were studied and resulted in a sensitive ECL sensor for hydrogen peroxide (H 2 O 2 ) that works in the concentration range of 1 nM to 0.9 μM. The scavenging effect of superoxide dismutase (SOD) on the H 2 O 2 electrode ECL was then exploited to design a biosensor for the determination of SOD in concentrations between 50 and 500 nM. (author)

  19. Development of luminol-N-hydroxyphthalimide chemiluminescence system for highly selective and sensitive detection of superoxide dismutase, uric acid and Co2.

    Science.gov (United States)

    Saqib, Muhammad; Qi, Liming; Hui, Pan; Nsabimana, Anaclet; Halawa, Mohamed Ibrahim; Zhang, Wei; Xu, Guobao

    2018-01-15

    N-hydroxyphthalimide (NHPI), a well known reagent in organic synthesis and biochemical applications, has been developed as a stable and efficient chemiluminescence coreactant for the first time. It reacts with luminol much faster than N-hydroxysuccinimide, eliminating the need of a prereaction coil used in N-hydroxysuccinimide system. Without using prereaction coil, the chemiluminescence peak intensities of luminol-NHPI system are about 102 and 26 times greater than that of luminol-N-hydroxysuccinimide system and classical luminol-hydrogen peroxide system, respectively. The luminol-NHPI system achieves the highly sensitive detection of luminol (LOD = 70pM) and NHPI (LOD = 910nM). Based on their excellent quenching efficiencies, superoxide dismutase and uric acid are sensitively detected with LODs of 3ng/mL and 10pM, respectively. Co 2+ is also detected a LOD of 30pM by its remarkable enhancing effect. Noteworthily, our method is at least 4 orders of magnitude more sensitive than previously reported uric acid detection methods, and can detect uric acid in human urine and Co 2+ in tap and lake water real samples with excellent recoveries in the range of 96.35-102.70%. This luminol-NHPI system can be an important candidate for biochemical, clinical and environmental analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Effect of gold nanoparticle as a novel nanocatalyst on luminol-hydrazine chemiluminescence system and its analytical application

    International Nuclear Information System (INIS)

    Safavi, A.; Absalan, G.; Bamdad, F.

    2008-01-01

    In this work the catalytic role of unsupported gold nanoparticles on the luminol-hydrazine reaction is investigated. Gold nanoparticles catalyze the reaction of hydrazine and dissolved oxygen to generate hydrogen peroxide and also catalyze the oxidation of luminol by the produced hydrogen peroxide. The result is an intense chemiluminescence (CL) due to the excited 3-aminophthalate anion. In the absence of gold nanoparticles no detectable CL was observed by the reaction of luminol and hydrazine unless an external oxidant is present in the system. The size effect of gold nanoparticles on the CL intensity was investigated. The most intensive CL signals were obtained with 15-nm gold nanoparticles. UV-vis spectra and transmission electron microscopy studies were used to investigate the CL mechanism. The luminol and hydroxide ion concentration, gold nanoparticles size and flow rate were optimized. The proposed method was successfully applied to the determination of hydrazine in boiler feed water samples. Between 0.1 and 30 μM of hydrazine could be determined with a detection limit of 30 nM

  1. Study on Enhancement Principle and Stabilization for the Luminol-H2O2-HRP Chemiluminescence System.

    Directory of Open Access Journals (Sweden)

    Lihua Yang

    Full Text Available A luminol-H2O2-HRP chemiluminescence system with high relative luminescent intensity (RLU and long stabilization time was investigated. First, the comparative study on the enhancement effect of ten compounds as enhancers to the luminol-H2O2-HRP chemiluminescence system was carried out, and the results showed that 4-(imidazol-1-ylphenol (4-IMP, 4-iodophenol (4-IOP, 4-bromophenol (4-BOP and 4-hydroxy-4'-iodobiphenyl (HIOP had the best performance. Based on the experiment, the four enhancers were dissolved in acetone, acetonitrile, methanol, and dimethylformamide (DMF with various concentrations, the results indicated that 4-IMP, 4-IOP, 4-BOP and HIOP dissolved in DMF with the concentrations of 0.2%, 3.2%, 1.6% and 3.2% could get the highest RLU values. Subsequently, the influences of pH, ionic strength, HRP, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol on the stabilization of the luminol-H2O2-HRP chemiluminescence system were studied, and we found that pH value, ionic strength, 4-IMP, 4-IOP, 4-BOP, HIOP, H2O2 and luminol have little influence on luminescent stabilization, while HRP has a great influence. In different ranges of HRP concentration, different enhancers should be selected. When the concentration is within the range of 0~6 ng/mL, 4-IMP should be selected. When the concentration of HRP ranges from 6 to 25 ng/mL, 4-IOP was the best choice. And when the concentration is within the range of 25~80 ng/mL, HIOP should be selected as the enhancer. Finally, the three well-performing chemiluminescent enhanced solutions (CESs have been further optimized according to the three enhancers (4-IMP, 4-IOP and HIOP in their utilized HRP concentration ranges.

  2. Automated determination of asulam by enhanced chemiluminescence using luminol/peroxidase system.

    Science.gov (United States)

    Sánchez, Francisco García; Díaz, Aurora Navas; Bracho, Visitación; Aguilar, Alfonso; Algarra, Manuel

    2009-01-01

    A flow injection system with chemiluminescence detection for the determination of asulam, enhancer of the system luminol-H(2)O(2)-horseradish peroxidase, is proposed. The method shows a moderate selectivity against other pesticides usually present in formulations of herbicides and in water. The procedure was applied to the determination of asulam in tap water samples and a recovery study was carried out in order to validate the method. The obtained results show acceptable recovery values (between 88.3 and 93.9%). The detection limit for asulam was 0.12 ng/mL. The precision of the method expressed as relative standard deviation was 1.55% (n = 8), at the 19 ng/mL level.

  3. Forensic Luminol Blood Test for Preventing Cross-contamination in Dentistry: An Evaluation of a Dental School Clinic.

    Science.gov (United States)

    Bortoluzzi, Marcelo Carlos; Cadore, Peterson; Gallon, Andrea; Imanishi, Soraia Almeida Watanabe

    2014-10-01

    More than 200 different diseases may be transmitted from exposure to blood in the dental setting. The aim of this study is to identify possible faults in the crosscontamination chain control in a dental school clinic searching for traces of blood in the clinical contact surfaces (CCS) through forensic luminol blood test. Traces of invisible blood where randomly searched in CCS of one dental school clinic. Forty eight surfaces areas in the CCS were tested and the presence of invisible and remnant blood was identified in 28 (58.3%) items. We suggest that the luminol method is suitable for identifying contamination with invisible blood traces and this method may be a useful tool to prevent cross-contamination in the dental care setting.

  4. Forensic Luminol Blood Test for Preventing Cross-contamination in Dentistry: An Evaluation of a Dental School Clinic

    OpenAIRE

    Marcelo Carlos Bortoluzzi; Peterson Cadore; Andrea Gallon; Soraia Almeida Watanabe Imanishi

    2014-01-01

    Background: More than 200 different diseases may be transmitted from exposure to blood in the dental setting. The aim of this study is to identify possible faults in the cross-contamination chain control in a dental school clinic searching for traces of blood in the clinical contact surfaces (CCS) through forensic luminol blood test. Methods: Traces of invisible blood where randomly searched in CCS of one dental school clinic. Results: Forty eight surfaces areas in the CCS were tes...

  5. Detecção de manchas de sangue pelo luminol onde houve entintamento das paredes – estudo de caso

    Directory of Open Access Journals (Sweden)

    Geraldo Elias Miranda

    2016-04-01

    Full Text Available O luminol é um teste presuntivo para detecção de manchas de sangue, muito sensível, efetivo e seletivo. O objetivo deste trabalho é avaliar a eficácia do luminol na detecção de sangue após o entintamento das paredes em um local onde ocorreu um homicídio. Os peritos se dirigiram para a casa do suspeito e com o uso de uma espátula metálica retiraram parte do revestimento da parede do quarto e após nova aplicação do luminol observaram a mancha branco-azulada, intensa e de duração típica de sangue latente. Os fragmentos retirados da parede foram enviados para o laboratório de DNA que confirmou tratar-se de sangue da vítima. A aplicação dessa técnica no caso em questão encontrando manchas de sangue sob a pintura ajudou a perícia a traçar a dinâmica do evento, ficando mais clara a posição da vítima quando foi alvejada com tiros pelo seu agressor além de ligar aquele local à vítima. O luminol não é capaz de detectar sangue que está sob a tinta da parede. A deteção do sangue latente somente ocorre quando a camada de tinta é retirada.

  6. A novel luminol chemiluminescent method catalyzed by silver/gold alloy nanoparticles for determination of anticancer drug flutamide.

    Science.gov (United States)

    Chaichi, Mohammad Javad; Azizi, Seyed Naser; Heidarpour, Maryam

    2013-12-01

    It was found that silver/gold alloy nanoparticles enhance the chemiluminescence (CL) of the luminol-H2O2 system in alkaline solution. The studies of UV-Vis spectra, CL spectra, effects of concentrations luminol, hydrogen peroxide and silver/gold alloy nanoparticles solutions were carried out to explore the CL enhancement mechanism. Flutamide was found to quench the CL signals of the luminol-H2O2 reaction catalyzed by silver/gold alloy nanoparticles, which made it applicable for the determination of flutamide. Under the optimum conditions, the CL intensity is proportional to the concentration of the flutamide in solution over the range 5.0 × 10(-7) to 1.0 × 10(-4)mol L(-1). Detection limit was obtained 1.2 × 10(-8)mol L(-1)and the relative standard deviation (RSD) γ5%. This work is introduced as a new method for the determination of flutamide in commercial tablets. Box-Behnken experimental design is applied to investigate and validate the CL measurement parameters. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Luminol functionalized gold nanoparticles as colorimetric and chemiluminescent probes for visual, label free, highly sensitive and selective detection of minocycline

    Science.gov (United States)

    He, Yi; Peng, Rufang

    2014-11-01

    In this work, luminol functionalized gold nanoparticles (LuAuNPs) were used as colorimetric and chemiluminescent probes for visual, label free, sensitive and selective detection of minocycline (MC). The LuAuNPs were prepared by simple one-pot reduction of HAuCl4 with luminol, which exhibited a good chemiluminescence (CL) activity owing to the presence of luminol molecules on their surface and surface plasmon resonance absorption. In the absence of MC, the color of LuAuNPs was wine red and their size was relatively small (˜25 nm), which could react with silver nitrate, producing a strong CL emission. Upon the addition of MC at acidic buffer solutions, the electrostatic interaction between positively charged MC and negatively charged LuAuNPs caused the aggregation of LuAuNPs, generating a purple or blue color. Simultaneously, the aggregated LuAuNPs did not effectively react with silver nitrate, producing a weak CL emission. The signal change was linearly dependent on the logarithm of MC concentration in the range from 30 ng to 1.0 μg for colorimetric detection and from 10 ng to 1.0 μg for CL detection. With colorimetry, a detection limit of 22 ng was achieved, while the detection limit for CL detection modality was 9.7 ng.

  8. Luminol functionalized gold nanoparticles as colorimetric and chemiluminescent probes for visual, label free, highly sensitive and selective detection of minocycline

    International Nuclear Information System (INIS)

    He, Yi; Peng, Rufang

    2014-01-01

    In this work, luminol functionalized gold nanoparticles (LuAuNPs) were used as colorimetric and chemiluminescent probes for visual, label free, sensitive and selective detection of minocycline (MC). The LuAuNPs were prepared by simple one-pot reduction of HAuCl 4 with luminol, which exhibited a good chemiluminescence (CL) activity owing to the presence of luminol molecules on their surface and surface plasmon resonance absorption. In the absence of MC, the color of LuAuNPs was wine red and their size was relatively small (∼25 nm), which could react with silver nitrate, producing a strong CL emission. Upon the addition of MC at acidic buffer solutions, the electrostatic interaction between positively charged MC and negatively charged LuAuNPs caused the aggregation of LuAuNPs, generating a purple or blue color. Simultaneously, the aggregated LuAuNPs did not effectively react with silver nitrate, producing a weak CL emission. The signal change was linearly dependent on the logarithm of MC concentration in the range from 30 ng to 1.0 μg for colorimetric detection and from 10 ng to 1.0 μg for CL detection. With colorimetry, a detection limit of 22 ng was achieved, while the detection limit for CL detection modality was 9.7 ng. (paper)

  9. Optical fiber chemiluminescence sensor for iron (II) ion based on immobilized luminol

    International Nuclear Information System (INIS)

    Alipao, Arthur A.; Sevilla, Fortunato III.

    1999-01-01

    A chemiluminescence (CL) sensor for iron (II) was developed based on the catalytic action of the analyte on the CL reaction between luminol and oxygen. The reagents were immobilized on a cellulose membrane and set on a reaction cell which was coupled by means of an optical fiber to a spectrofluorometer. The concentration of iron(II) was quantified by measuring the intensity of the light generated from the CL reaction. The response of the sensor system was rapid and highly reproducible. Good sensitivity was displayed by the sensor system over the five orders of magnitude of iron(II) ion concentration. The calibration curve consisted of two portions: (1) a linear range at lower concentrations (7.5 x 10 -7 M to 1.0 x 10 -4 M) exhibiting a positive slope arising from a catalytic action, and (2) a linear range at higher concentrations (5.0 x 10 -2 M to 2.5 x 10 -4 M) wherein the slope is negative due to an inhibitory action of iron(II) on the CL reaction. The sensor system was highly selective for iron(II) ions. (Author)

  10. Luminol-dependent chemiluminescence in antibody-sensitized neutrophils stimulated with protein A-bearing staphylococci.

    Science.gov (United States)

    Nishihara, S; Seki, K; Ikigai, H; Masuda, S

    1988-01-01

    When mouse polymorphonuclear leukocytes (PMNs) sensitized with rabbit antibody to mouse Ehrlich ascites tumor cells were stimulated by Staphylococcus aureus Cowan I cells, a conspicuous luminol-dependent chemiluminescence was observed in the absence of opsonin. The profile of the chemiluminescence (CL) response evoked by staphylococcal cells from antibody-sensitized PMNs had two peaks. An initial peak, observed within 1 min after stimulation, was sharp and high and a second peak, observed about 5 min after stimulation, was low and extended. The CL response of antibody-sensitized PMNs stimulated by S. aureus Cowan I cells was dose-dependently blocked by preincubation with soluble SpA. Cells of a mutant derived from S. aureus Cowan I strain with trace amounts of cell-bound SpA failed to stimulate the antibody-sensitized PMNs to generate the CL response. The antibody-sensitized PMNs were found to phagocytize SpA-bearing S. aureus cells even in the absence of opsonic serum. These results suggest that the observation presented here might provide a useful tool for the investigation of CL response of PMNs.

  11. Development and application of a luminol-based nitrogen dioxide detector

    International Nuclear Information System (INIS)

    Wendel, G.J.

    1985-01-01

    An instrument for the continuous measurement of nitrogen dioxide (NO 2 ) at all atmospheric concentration ranges and conditions was developed. The detector is based on the chemiluminescent reaction between 5-amino-2,3-dihydro-1,4-phthalazinedione (luminol) and NO 2 in alkaline aqueous solution. Development included the optimization of the cell design and the solution composition. Sodium sulfite (Na 2 SO 3 ) and methanol (CH 3 OH) were added to the solution to improve sensitivity and specificity. The detector was favorably compared to two different instruments measuring NO 2 by NO + O 3 chemiluminescent and by a tunable diode laser absorption spectrometry system. The detector has demonstrated a detection limit of 30 parts-per-trillion by volume (ppt) and a frequency response of 0.3 Hz. The instrument was operated for two one-month periods on Bermuda. The purpose was to study air masses from the East Coast of the United States after transport over the ocean. Average daily values were 400 ppt with values as low as 100 ppt measured. Other field experiments involved monitoring of NO 2 in ambient air in the range of 1 to 60 parts-per-billion by volume

  12. Luminol-based electrochemiluminescent biosensors for highly sensitive medical diagnosis and rapid antioxidant detection

    Science.gov (United States)

    Tamiya, Eiichi; Inoue, Yuki; Saito, Masato

    2018-03-01

    We present a review of luminol-based electrochemiluminescence (ECL) biosensors that perform enzymatic reactions and bioanalysis using antioxidant molecules by controlling the spatiotemporal production of a luminescent substrate, catalase activity, and glycated albumin (GA). The ECL intensity depends on the antioxidant capacity because radicals are neutralized by the antioxidants, which suppresses the luminescence. The antioxidant capacities of 22 types beverages were evaluated by comparison with a standard curve of Trolox. The time necessary for the ECL measurement of the antioxidant capacity is only 2 min with screen-printed electrodes and a portable ECL measurement system. Our system was also used to monitor reactive oxygen species released from neutrophils, which play an important role in the immune system, defending against viral and bacterial infections. The quenching of ECL imaging by catalase reaction localized in the multichamber electrode using the electrogenerated substrate was examined as a potential candidate for a sensitive reporting system. The substrate was successfully generated at applied potentials between -1 and -0.4 V in multichamber electrodes and the substrate confinement within the chamber was observed within 60 s of generation. The microchamber electrode system demonstrated a detection limit of 90 fM catalase. We also demonstrated a detection limit of 0.1 µM GA in human serum albumin, which is an improvement of about 70 times over colorimetric methods.

  13. Luminol Chemiluminescence Catalyzed by Silver Nanoparticles for the Sensitive Determination of Penicillamine

    Directory of Open Access Journals (Sweden)

    Fakhr Eldin O. Suliman

    2018-01-01

    Full Text Available A sequential injection method for the determination of penicillamine (PA was developed based on quenching the chemiluminescence generated by oxidation of luminol by hydrogen peroxide in presence of silver nanoparticles (AgNPs. The chemiluminescence (CL of the reaction was found to greatly enhance in presence of AgNPs due to the increased catalyst surface area. The method was sensitive and found suitable for analysis of penicillamine in pharmaceutical preparations. Linear calibration curve is obtained in the range 0.2-1.0 mg mL-1 with a relative standard deviation less than 2%. A recovery percent of 102.3 ±0.2 was obtained with the tablets matrix indicating reasonable selectivity of the method for PA in tablets. The mechanism of quenching of the CL reaction was investigated by UV-Visible spectroscopy and transmission electron microscopy as well as by theoretical calculations using DFT-B3LYP method. The covalent attachment of PA to the AgNPs triggers aggregation of the particles thereby diminishing the surface significantly. The method was applied for the assay of PA in pharmaceutical preparations.

  14. Effect of the luminol signal enhancer on the chemiluminescence intensity and kinetics

    International Nuclear Information System (INIS)

    Liang Yanli; Yu Fei; Yu Songcheng; Wu Yongjun; Zhang Hongquan; Qu Lingbo

    2012-01-01

    The novel p-phenol derivatives, 4-(1-imidazolyl)-phenol, 4-hydroxybiphenyl, 4-hydroxy-4′-iodobiphenyl were employed as highly potent signal enhancers of luminol-hydrogen peroxide (H 2 O 2 )-horseradish peroxidase (HRP) chemiluminescence (CL) system. The CL reaction conditions were optimized, and the enhancement characteristics of these enhancers were compared with each other. The employment of these molecules greatly affected important assay parameters. Practically, the use of a novel enhancer, even a slightly change of the structure (or concentration) of 4-substituted phenol derivative, could affect assay properties quite dramatically. Furthermore, the use of different enhancers in the luminol–H 2 O 2 –HRP system can affect not only the intensity of the CL signal, which is well known, but also its kinetics. The experiment data indicated that the stronger intensity was combined with a more rapid decrease of the CL signal. - Highlights: ► 4-IMP, 4-BIP and HIOP take on different signal enhancement and light kinetics. ► The employment of 4-BIP resulted in a significant improvement of the detection limit. ► The magnitude of the signal enhancement was about 2.5-fold in the same system.

  15. Quantitative Monitoring of Cefradine in Human Urine Using a Luminol/Sulfobutylether-β-Cyclodextrin Chemiluminescence System

    Science.gov (United States)

    Shen, M. X.; Tan, X. J.; Song, Zh. H.

    2018-05-01

    In this paper, a sensitive, rapid, and simple flow-injection chemiluminescence (FI-CL) technique is described for determining cefradine in human urine and capsule samples at the picogram level. The results show that cefradine within 0.1-100.0 nmol/L quantitatively quenches the CL intensity of the luminol/sulfo butylether-β-cyclodextrin (SBE-β-CD) system, with a relative correlation coefficient r of 0.9931. Subsequently, the possible mechanism for the quenching phenomenon is discussed in detail using the FI-CL and molecular docking methods. The proposed CL method, with a detection limit of 0.03 nmol/L (3σ) and relative standard deviations administration, the cefradine reaches a maximum value of 1.37 ± 0.02 mg/mL at 2.0 h in urine, and the total excretion is 4.41 ± 0.03 mg/mL within 8.0 h. The absorption rate constant ka, the elimination rate constant ke, and the half-life t1/2 are 0.670 ± 0.008 h-1, 0.744 ± 0.005 h-1, and 0.93 ± 0.05 h, respectively.

  16. Quantitative Monitoring of Cefradine in Human Urine Using a Luminol/Sulfobutylether-β-Cyclodextrin Chemiluminescence System

    Science.gov (United States)

    Shen, M. X.; Tan, X. J.; Song, Zh. H.

    2018-05-01

    In this paper, a sensitive, rapid, and simple flow-injection chemiluminescence (FI-CL) technique is described for determining cefradine in human urine and capsule samples at the picogram level. The results show that cefradine within 0.1-100.0 nmol/L quantitatively quenches the CL intensity of the luminol/sulfo butylether-β-cyclodextrin (SBE-β-CD) system, with a relative correlation coefficient r of 0.9931. Subsequently, the possible mechanism for the quenching phenomenon is discussed in detail using the FI-CL and molecular docking methods. The proposed CL method, with a detection limit of 0.03 nmol/L (3σ) and relative standard deviations 3.0% (N = 7), is then implemented to monitor the excretion of cefradine in human urine. After orally administration, the cefradine reaches a maximum value of 1.37 ± 0.02 mg/mL at 2.0 h in urine, and the total excretion is 4.41 ± 0.03 mg/mL within 8.0 h. The absorption rate constant ka, the elimination rate constant ke, and the half-life t1/2 are 0.670 ± 0.008 h-1, 0.744 ± 0.005 h-1, and 0.93 ± 0.05 h, respectively.

  17. The comparative performance of PMI estimation in skeletal remains by three methods (C-14, luminol test and OHI): analysis of 20 cases.

    Science.gov (United States)

    Cappella, Annalisa; Gibelli, Daniele; Muccino, Enrico; Scarpulla, Valentina; Cerutti, Elisa; Caruso, Valentina; Sguazza, Emanuela; Mazzarelli, Debora; Cattaneo, Cristina

    2015-01-27

    When estimating post-mortem interval (PMI) in forensic anthropology, the only method able to give an unambiguous result is the analysis of C-14, although the procedure is expensive. Other methods, such as luminol tests and histological analysis, can be performed as preliminary investigations and may allow the operators to gain a preliminary indication concerning PMI, but they lack scientific verification, although luminol testing has been somewhat more accredited in the past few years. Such methods in fact may provide some help as they are inexpensive and can give a fast response, especially in the phase of preliminary investigations. In this study, 20 court cases of human skeletonized remains were dated by the C-14 method. For two cases, results were chronologically set after the 1950s; for one case, the analysis was not possible technically. The remaining 17 cases showed an archaeological or historical collocation. The same bone samples were also screened with histological examination and with the luminol test. Results showed that only four cases gave a positivity to luminol and a high Oxford Histology Index (OHI) score at the same time: among these, two cases were dated as recent by the radiocarbon analysis. Thus, only two false-positive results were given by the combination of these methods and no false negatives. Thus, the combination of two qualitative methods (luminol test and microscopic analysis) may represent a promising solution to cases where many fragments need to be quickly tested.

  18. Revelado de manchas latentes: efectividad del luminol y evaluación de su efecto sobre el estudio del DNA Development of latent stains: effectiveness of luminol and evaluation of its effect on DNA analysis

    OpenAIRE

    A. Castelló Ponce; M. Álvarez Seguí; M. Miquel Feucht; F.A. Verdú Pascual

    2002-01-01

    Con frecuencia, las ropas manchadas de sangre durante un acto violento son lavadas para tratar de eliminar los vestigios. El análisis del DNA mediante PCR ha dotado a la criminalística de la posibilidad de estudiar indicios mínimos que, con otros métodos, sería imposible analizar. Este tipo de muestra plantea una mayor dificultad en la detección de la mancha. En el presente trabajo se analiza la sensibilidad del luminol para encontrar manchas de sangre invisibles. Sobre estas manchas se inten...

  19. Highly luminescent S,N co-doped carbon quantum dots-sensitized chemiluminescence on luminol-H2 O2 system for the determination of ranitidine.

    Science.gov (United States)

    Chen, Jianqiu; Shu, Juan; Chen, Jiao; Cao, Zhiran; Xiao, An; Yan, Zhengyu

    2017-05-01

    S,N co-doped carbon quantum dots (N,S-CQDs) with super high quantum yield (79%) were prepared by the hydrothermal method and characterized by transmission electron microscopy, photoluminescence, UV-Vis spectroscopy and Fourier transformed infrared spectroscopy. N,S-CQDs can enhance the chemiluminescence intensity of a luminol-H 2 O 2 system. The possible mechanism of the luminol-H 2 O 2 -(N,S-CQDs) was illustrated by using chemiluminescence, photoluminescence and ultraviolet analysis. Ranitidine can quench the chemiluminescence intensity of a luminol-H 2 O 2 -N,S-CQDs system. So, a novel flow-injection chemiluminescence method was designed to determine ranitidine within a linear range of 0.5-50 μg ml -1 and a detection limit of 0.12 μg ml -1 . The method shows promising application prospects. Copyright © 2016 John Wiley & Sons, Ltd.

  20. Electrochemiluminescence of luminol enhanced by the synergetic catalysis of hemin and silver nanoparticles for sensitive protein detection.

    Science.gov (United States)

    Jiang, Xinya; Chai, Yaqin; Wang, Haijun; Yuan, Ruo

    2014-04-15

    A novel and ultrasensitive electrochemiluminescence (ECL) immunosensor, which was based on the amplifying ECL of luminol by hemin-reduced graphene oxide (hemin-rGO) and Ag nanoparticles (AgNPs) decorated reduced graphene oxide (Ag-rGO), was constructed for the detection of carcinoembryonic antigen (CEA). For this proposed sandwich-type ECL immunosensor, Au nanoparticles electrodeposited (DpAu) onto hemin-rGO (DpAu/hemin-rGO) constructed the base of the immunosensor. DpAu had outstanding electrical conductivity to promote the electron transfer at the electrode interface and had good biocompatibility to load large amounts of primary antibody (Ab1), which provided an excellent platform for this immunosensor. Moreover, AgNPs and glucose oxidase (GOD) functionalized graphene labeled secondary antibody (Ag-rGO-Ab2-GOD) was designed as the signal probe for the sandwiched immunosensor. Not only did the hemin-rGO improve the electron transfer of the electrode surface, but hemin also further amplified the ECL signal of luminol in the presence of hydrogen peroxide (H2O2). With the aid of Ag-rGO-Ab2-GOD, enhanced signal was obtained by in situ generation of H2O2 and catalysis of AgNPs to ECL reaction of the luminol-H2O2 system. The as-prepared ECL immunosensor exhibited excellent analytical property for the detection of CEA in the range from 0.1 pg mL(-1) to 160 ng mL(-1) with a detection limit of 0.03 pg mL(-1) (SN(-1)=3). © 2013 Published by Elsevier B.V.

  1. Flow Injection Photosensitized Chemiluminescence of Luminol with Cu(II-Rose Bengal: Mechanistic Approach and Vitamin A and C Determination

    Directory of Open Access Journals (Sweden)

    Muhammad Asgher

    2014-01-01

    Full Text Available Rose Bengal photosensitized flow injection chemiluminescence method is reported using luminol-Cu(II for the determination of vitamins A and C in pharmaceutical formulations. The reaction is based on the enhancement effect of analyte in the production of anion radicals of Rose Bengal (RB•− which rapidly interact with dissolved oxygen and generate superoxide anions radicals (O2•− and hydrogen peroxide (H2O2. Highly reactive hydroxyl radicals (•OH were produced via dismutation of H2O2 by catalyst (Cu2+. The generated superoxide anions radicals and hydroxyl radicals thus oxidize luminol in alkaline medium to generate strong chemiluminescence. The limit of detection (3s of the blank, n=6 of vitamins A and C and RB was found to be 0.008, 0.005, and 0.05 μg mL−1, respectively. The sample throughput of 70 h−1 for vitamins A and C and 30 h−1 for RB was found. Calibration curve was linear in the range of 0.05–15, 0.01–20, and 0.1–50 μg mL−1 for vitamins A and C and RB, respectively, with relative standard deviations (RSDs; n=3 in the range 1.6–3.6%. The method was successfully applied to pharmaceutical formulations and the results obtained were in good agreement with the labeled values.

  2. Enhancement effect of CdTe quantum dots-IgG bioconjugates on chemiluminescence of luminol-H2O2 system

    International Nuclear Information System (INIS)

    Kanwal, Shamsa; Traore, Zoumana; Zhao Chunfang; Su Xingguang

    2010-01-01

    In this paper we developed an entirely new and highly sensitive luminol-H 2 O 2 flow injection chemiluminescence system using the enhancement effect of CdTe quantum dots-IgG bioconjugates. Immunoglobulin G (IgG) as a kind of bio-molecule was conjugated to different sized CdTe semiconductor quantum dots (QDs). Using PL spectra and CL intensity profiles, it was found that chemiluminescence resonance energy transfer (CRET) was possibly occurring between CdTe-IgG bioconjugate and luminol. Under optimum conditions, increase of IgG concentration in CdTe-IgG bioconjugate resulted enhancing effect on CL intensity of luminol-H 2 O 2 system. Moreover quenching effects on CL intensity by addition of different proteases can construct turn off biosensor for these proteases with low detection limits and wide linear range. Furthermore, the effects of various organic and inorganic species on CdTe-IgG bioconjugates enhanced luminol-H 2 O 2 CL system were also studied in this paper.

  3. A turn-on chemiluminescence biosensor for selective and sensitive detection of adenosine based on HKUST-1 and QDs-luminol-aptamer conjugates.

    Science.gov (United States)

    Lin, Yanna; Dai, Yuxue; Sun, Yuanling; Ding, Chaofan; Sun, Weiyan; Zhu, Xiaodong; Liu, Hao; Luo, Chuannan

    2018-05-15

    In this work, HKUST-1 and QDs-luminol-aptamer conjugates were prepared. The QDs-luminol-aptamer conjugates can be adsorbed by graphene oxide through π-π conjugation. When the adenosine was added, the QDs-luminol-aptamer conjugates were released from magnetic graphene oxide (MGO), the chemiluminescent switch was turned on. It was reported that HKUST-1 can catalyze the chemiluminescence reaction of luminol-H 2 O 2 system in an alkaline medium, and improve the chemiluminescence resonance energy transfer (CRET) between chemiluminescence and QDs indirectly. Thus, the adenosine can be detected sensitively. Based on this phenomenon, the excellent platform for detection of adenosine was established. Under the optimized conditions, the linear detection range for adenosine was 1.0 × 10 -12 -2.2 × 10 -10 mol/L with a detection limit of 2.1 × 10 -13 mol/L. The proposed method was successfully used for adenosine detection in biological samples. Copyright © 2018 Elsevier B.V. All rights reserved.

  4. Electrochemiluminescence quenching of luminol by CuS in situ grown on reduced graphene oxide for detection of N-terminal pro-brain natriuretic peptide.

    Science.gov (United States)

    Li, Xiaojian; Lu, Peng; Wu, Bin; Wang, Yaoguang; Wang, Huan; Du, Bin; Pang, Xuehui; Wei, Qin

    2018-07-30

    A novel electrochemiluminescence (ECL) signal-off strategy based on CuS in situ grown on reduced graphene oxide (CuS-rGO) quenching luminol/H 2 O 2 system was firstly proposed. Luminol was grafted on the surface of Au@Fe 3 O 4 -Cu 3 (PO 4 ) 2 nanoflowers (Luminol-Au@Fe 3 O 4 -Cu 3 (PO 4 ) 2 ) which exhibited excellent catalytic effect towards the reduction of H 2 O 2 to enhance the ECL intensity of luminol. Cu 3 (PO 4 ) 2 nanoflowers showed large surface area which can immobilize more Fe 3 O 4 and Au nanoparticles. The quenching mechanism of CuS-rGO was due to ECL resonance energy transfer (RET). The spectral overlap between fluorescence spectrum of Luminol-Au@Fe 3 O 4 -Cu 3 (PO 4 ) 2 and UV-vis absorption spectrum of CuS-rGO revealed that resonance energy transfer was possible. Au nanoparticles were immobilized on the surface of CuS-rGO to capture secondary antibodies. After a sandwich-type immunoreaction, a remarkable decrease of ECL signal was observed. Under the optimal conditions, the immunosensor showed excellent performance for N-terminal pro-brain natriuretic peptide (NT-proBNP) detection with a wide detection range from 0.5 pg mL -1 to 20 ng mL -1 and a low detection limit of 0.12 pg mL -1 (S/N = 3). The prepared NT-proBNP immunosensor displayed high sensitivity, excellent stability and good specificity. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Long-term chemiluminescence signal is produced in the course of luminol oxidation catalyzed by enhancer-independent peroxidase purified from Jatropha curcas leaves.

    Science.gov (United States)

    Duan, Peipei; Cai, Feng; Luo, Yongting; Chen, Yangxi; Zou, Shujuan

    2015-09-01

    Isoenzyme c of horseradish peroxidase (HRP-C) is widely used in enzyme immunoassay combined with chemiluminescence (CL) detection. For this application, HRP-C activity measurement is usually based on luminol oxidation in the presence of hydrogen peroxide (H2O2). However, this catalysis reaction was enhancer dependent. In this study, we demonstrated that Jatropha curcas peroxidase (JcGP1) showed high efficiency in catalyzing luminol oxidation in the presence of H2O2. Compared with HRP-C, the JcGP1-induced reaction was enhancer independent, which made the enzyme-linked immunosorbent assay (ELISA) simpler. In addition, the JcGP1 catalyzed reaction showed a long-term stable CL signal. We optimized the conditions for JcGP1 catalysis and determined the favorable conditions as follows: 50 mM Tris buffer (pH 8.2) containing 10 mM H2 O2, 14 mM luminol and 0.75 M NaCl. The optimum catalysis temperature was 30°C. The detection limit of JcGP1 under optimum condition was 0.2 pM. Long-term stable CL signal combined with enhancer-independent property indicated that JcGP1 might be a valuable candidate peroxidase for clinical diagnosis and enzyme immunoassay with CL detection. Copyright © 2014 John Wiley & Sons, Ltd.

  6. Strong enhancement of the electrochemiluminescence of luminol by AuAg and PtAg alloy nanoclusters, and its sensitization by phenolic artificial oestrogens

    International Nuclear Information System (INIS)

    Wang, Ke; Tu, Yifeng; Wei, Xiuhua

    2014-01-01

    This paper reports on the synthesis of AuAg and PtAg alloy nanoclusters (NCs) and their enhancement effect on the electrochemiluminescence (ECL) of luminol. The conditions of synthesis were optimized, and the structure and properties of the NCs were characterized by X-ray diffraction, transmission electron microscopy, electrochemistry, and optical spectroscopy. The NCs are found to intensify (by up to 20 times) the ECL of luminol in solution of pH 8.5. This finding can largely extend the useful pH range of the ECL of luminol. The enhanced ECL is strongly affected by oxygen and hydrogen peroxide, and the mechanism of enhancement is attributed to the accelerated production of reactive oxygen species. The enhanced ECL is also affected by phenolic artificial estrogens, and this was used for their determination with detection limits as low as 700 pg L −1 (with AuAg) and 1.6 ng L −1 (with PtAg). The method was applied to the determination of such estrogens in egg samples using diethylstilbestrol as a reference substance. (author)

  7. Luminol testing in detecting modern human skeletal remains: a test on different types of bone tissue and a caveat for PMI interpretation.

    Science.gov (United States)

    Caudullo, Giorgio; Caruso, Valentina; Cappella, Annalisa; Sguazza, Emanuela; Mazzarelli, Debora; Amadasi, Alberto; Cattaneo, Cristina

    2017-01-01

    When forensic pathologists and anthropologists have to deal with the evaluation of the post-mortem interval (PMI) in skeletal remains, luminol testing is frequently performed as a preliminary screening method. However, the repeatability of this test on the same bone, as well as comparative studies on different bones of the same individual, has never been performed. Therefore, with the aim of investigating the influence that different types of bones may exert on the response to the luminol test, the present study analysed three different skeletal elements (femoral diaphysis, vertebra and cranial vault), gathered from ten recent exhumed skeletons (all with a 20-year PMI). The analysis was performed twice on the same bone after 2 months: the analysis at time 0 concerned the whole bone, whereas the second concerned only a part of the same bone taken during the first test (which already had been broken). The overall results showed different responses, depending on the type of bone and on the integrity of the samples. Negative results at the first analysis (6.6% out of the total of samples) are consistent with what is reported in the literature, whilst at the second analysis, the increase of about 20% of false-negative results highlights that the luminol test ought to be performed with caution in case of broken bones or elements which are taphonomically altered. Results have thus proven that the exposition to environmental agents might result in haemoglobin (Hb) loss, as detected even after only 2 months. The study also focused on the crucial issue of the type of bone subjected to testing, remarking the suitability of the femoral diaphysis (100% of positive responses at the first analysis vs only 18% of false-negative results at the second test, corresponding to 5% of total false-negative results) as opposed to other bone elements that showed a low yield. In particular, the cranial vault gave poor results, with 40% of discrepancy between results from the two analyses

  8. СHANGES IN PARAMETERS OF LUMINOL-DEPENDENT AND LUCIGENIN-DEPENDENT CHEMILUMINESCENCE OF PERIPHERAL BLOOD NEUTROPHILS IN PATIENTS WITH BLADDER CANCER IN THE DISEASE DYNAMICS

    Directory of Open Access Journals (Sweden)

    L. M. Kurtasova

    2015-01-01

    Full Text Available The study deals with parameters of luminol-dependent and lucigenin-dependent chemiluminescence (CL of peripheral blood neutrophils from patients with bladder cancer (BC prior to surgical treatment. We examined sixty patients (45 to 55 years old with advanced bladder cancer (TNM prior to the operation, and forty-six patients at 10 days after surgical treatment. A control group consisted of 56 healthy donors. Luminol-dependent and lucigenin-dependent chemiluminescence of blood neutrophils was assessed according to De Sole et al. (1983. Chemiluminescence assays of peripheral blood neutrophils from the patients with bladder cancer revealed changes in production of reactive oxygen species (ROS, both for initial stage of oxidation reaction, and total level of active oxygen radicals. We have found disturbed values of primary-to-secondary ROS ratio in the cells. In the patients with bladder cancer, some changes in oxidative metabolism of the blood neutrophils have been registered. These alterations may play an important role in promotion of potential effector cell functions, thus, probably, affecting the whole-scale development of a cytopathic effect exerted by neutrophilic granulocytes. 

  9. Multiple enhancement of luminol electrochemiluminescence using electrodes functionalized with titania nanotubes and platinum black: ultrasensitive determination of hydrogen peroxide, resveratrol, and dopamine

    International Nuclear Information System (INIS)

    Ming, Liang; Peng, Tingting; Tu, Yifeng

    2016-01-01

    We describe a substantial improvement of the electrochemiluminescence (ECL) of luminol which is widely used in flow injection analysis (FIA). It is based on synchronous dual sensitization of ECL by using titania nanotubes (TiNTs) and platinum black (PB). A piece of indium tin oxide (ITO) glass functionalized with TiNTs acts as the first working electrode, and a PB-modified platinum plate serves as the second one. By applying two constant potentials to the two electrodes, strong and consecutive ECL emission of luminol is obtained. The system works well in assays as shown for the successful quantitation of hydrogen peroxide (H 2 O 2 ), of the antioxidant resveratrol, and of the neutrotransmitter dopamine (DA) in spiked human serum samples. The detection limits for these three species (at a signal-to-noise ratio of 3) are as low as 66 pM (H 2 O 2 ), 22 nM (resveratrol), and 30 nM (DA). Recoveries in assays of DA in spiked serum range from 97.3 to 105.4 %. In our perception, the technique of dual sensitization represents a substantial improvement of the detection limits of ECL assays. (author)

  10. Enhancing and inhibiting effects of aromatic compounds on luminol-dimethylsulfoxide-OH(-) chemiluminescence and determination of intermediates in oxidative hair dyes by HPLC with chemiluminescence detection.

    Science.gov (United States)

    Zhou, Jian; Xu, Hong; Wan, Guo-Hui; Duan, Chun-Feng; Cui, Hua

    2004-10-08

    The effect of 36 aromatic compounds on the luminol-dimethylsulfoxide-OH(-) chemiluminescence (CL) was systematically studied. It was found that dihydroxybenzenes, and ortho- and para-substituted aminophenols and phenylenediamines inhibited the CL and phenols with three or more than three hydroxyls except phloroglucin tended to enhance the CL. The CL inhibition and enhancement was proposed to be dependent on whether superoxide anion radical (O(2)(-)) was competitively consumed by compounds in the CL system. Trihydroxybenzenes were capable of generating superoxide anion radical, leading to the CL enhancement, whereas dihydroxybenzenes were superoxide anion radical scavenger, causing the CL inhibition. Based on the inhibited CL, a novel method for the simultaneous determination of p-phenylenediamine, o-phenylenediamine, p-aminophenol, o-aminophenol, resorcinol and hydroquinone by high-performance liquid chromatography coupled with chemiluminescence detection was developed. The method has been successfully applied to determine intermediates in oxidative hair dyes and wastewater of shampooing after hair dyed.

  11. Effect of Human and Bovine Serum Albumin on kinetic Chemiluminescence of Mn (III-Tetrakis (4-Sulfonatophenyl Porphyrin-Luminol-Hydrogen Peroxide System

    Directory of Open Access Journals (Sweden)

    Sayed Yahya Kazemi

    2012-01-01

    Full Text Available The present work deals with an attempt to study the effect of human and bovine serum albumin on kinetic parameters of chemiluminescence of luminol-hydrogen peroxide system catalyzed by manganese tetrasulfonatophenyl porphyrin (MnTSPP. The investigated parameters involved pseudo-first-order rise and fall rate constant for the chemiluminescence burst, maximum level intensity, time to reach maximum intensity, total light yield, and values of the intensity at maximum CL which were evaluated by nonlinear least square program KINFIT. Because of interaction of metalloporphyrin with proteins, the CL parameters are drastically affected. The systems resulted in Stern-Volmer plots with values of 3.17×105 and 3.7×105M−1 in the quencher concentration range of 1.5×10−6 to 1.5×10−5 M for human serum albumin (HSA and bovine serum albumin (BSA, respectively.

  12. Luminol/CdTe quantum dots/sodium periodate system in conjunction with response-surface methodology for chemiluminometric determination of some tetracyclines

    International Nuclear Information System (INIS)

    Imani-Nabiyyi, Amin; Sorouraddin, Mohammad H.; Amjadi, Mohammad; Naseri, Abdolhossein

    2014-01-01

    A simple and sensitive chemiluminometric method is described for the determination of tetracycline (TC) and oxytetracycline (OTC) in their hydrochloride form on the basis of drastic enhancement of the chemiluminescence (CL) from the luminol/L-cysteine-capped CdTe quantum dots (QDs)/sodium periodate system, caused by the presence of TC and OTC hydrochloride. The method uses pH value at which tetracyclines have proven to be stable. Response surface methodology was exploited for optimizing the experimental conditions. The synthesized QDs were characterized by means of X-ray diffraction (XRD), transmission electron microscopy (TEM), UV–vis spectroscopy, and photoluminescence spectroscopy. A possible mechanism of the CL system was proposed based on the CL spectra and the effect of some specific radical-scavengers on the system. Under the optimum conditions linear dynamic ranges of 5.0×10 −8 –6.0×10 −6 mol L −1 and 5.0×10 −8 –8.0×10 −6 mol L −1 were observed for TC and OTC, respectively with corresponding detection limits of 2.2×10 −8 mol L −1 and 3.0×10 −8 mol L −1 . The precision (RSD%) for five replicate determinations of TC and OTC were respectively no more than 3.4% and 4.0%. The method was satisfactorily applied to the determination of TC.HCl and OTC.HCl in water samples, pharmaceutical formulations, and honey. - Highlights: • A simple, rapid, and sensitive method for determination of some tetracyclines. • Enhancement of CL of luminol/Cys-capped CdTe QDs/NaIO 4 with TC.HCl and OTC.HCl. • No need to chemical, photochemical, or any peculiar pretreatment of the samples. • Measuring the analytes in conditions at which they are stable. • Capable of measuring TCs in pharmaceutical, water, and honey samples

  13. Comparative study of ß-glucan induced respiratory burst measured by nitroblue tetrazolium assay and real-time luminol-enhanced chemiluminescence assay in common carp (Cyprinus carpio L.)

    DEFF Research Database (Denmark)

    Jiménez, Natalia Ivonne Vera; Pietretti, D.; Wiegertjes, G. F.

    2013-01-01

    kidney cells of carp. However, whereas the NBT assay was shown to detect the production of only superoxide anions, the real-time luminol-enhanced assay could detect the production of both superoxide anions and hydrogen peroxide. Only the chemiluminescence assay could reliably record the production of ROS......-point measurement based on the intracellular reduction of nitroblue tetrazolium (NBT) and a real-time luminol-enhanced assay based on the detection of native chemiluminescence. Both assays allowed for detection of dose-dependent changes in magnitude of the respiratory burst response induced by β-glucans in head...... on a real-time scale at frequent and continual time intervals for time course experiments, providing more detailed information on the respiratory burst response. The real-time chemiluminescence assay was used to measure respiratory burst activity in macrophage and neutrophilic granulocyte-enriched head...

  14. Luminol/CdTe quantum dots/sodium periodate system in conjunction with response-surface methodology for chemiluminometric determination of some tetracyclines

    Energy Technology Data Exchange (ETDEWEB)

    Imani-Nabiyyi, Amin; Sorouraddin, Mohammad H., E-mail: soruraddin@tabrizu.ac.ir; Amjadi, Mohammad; Naseri, Abdolhossein

    2014-07-01

    A simple and sensitive chemiluminometric method is described for the determination of tetracycline (TC) and oxytetracycline (OTC) in their hydrochloride form on the basis of drastic enhancement of the chemiluminescence (CL) from the luminol/L-cysteine-capped CdTe quantum dots (QDs)/sodium periodate system, caused by the presence of TC and OTC hydrochloride. The method uses pH value at which tetracyclines have proven to be stable. Response surface methodology was exploited for optimizing the experimental conditions. The synthesized QDs were characterized by means of X-ray diffraction (XRD), transmission electron microscopy (TEM), UV–vis spectroscopy, and photoluminescence spectroscopy. A possible mechanism of the CL system was proposed based on the CL spectra and the effect of some specific radical-scavengers on the system. Under the optimum conditions linear dynamic ranges of 5.0×10{sup −8}–6.0×10{sup −6} mol L{sup −1} and 5.0×10{sup −8}–8.0×10{sup −6} mol L{sup −1} were observed for TC and OTC, respectively with corresponding detection limits of 2.2×10{sup −8} mol L{sup −1} and 3.0×10{sup −8} mol L{sup −1}. The precision (RSD%) for five replicate determinations of TC and OTC were respectively no more than 3.4% and 4.0%. The method was satisfactorily applied to the determination of TC.HCl and OTC.HCl in water samples, pharmaceutical formulations, and honey. - Highlights: • A simple, rapid, and sensitive method for determination of some tetracyclines. • Enhancement of CL of luminol/Cys-capped CdTe QDs/NaIO{sub 4} with TC.HCl and OTC.HCl. • No need to chemical, photochemical, or any peculiar pretreatment of the samples. • Measuring the analytes in conditions at which they are stable. • Capable of measuring TCs in pharmaceutical, water, and honey samples.

  15. Investigation of iron(III) reduction and trace metal interferences in the determination of dissolved iron in seawater using flow injection with luminol chemiluminescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Ussher, Simon J. [School of Earth, Ocean and Environmental Sciences (SEOES), University of Plymouth, Drake Circus, Plymouth PL4 8AA (United Kingdom); Milne, Angela [School of Earth, Ocean and Environmental Sciences (SEOES), University of Plymouth, Drake Circus, Plymouth PL4 8AA (United Kingdom); Department of Oceanography, Florida State University, Tallahassee, FL 32306-4320 (United States); Landing, William M. [Department of Oceanography, Florida State University, Tallahassee, FL 32306-4320 (United States); Attiq-ur-Rehman, Kakar [Department of Chemistry, University of Balochistan, Quetta (Pakistan); Seguret, Marie J.M.; Holland, Toby [School of Earth, Ocean and Environmental Sciences (SEOES), University of Plymouth, Drake Circus, Plymouth PL4 8AA (United Kingdom); Achterberg, Eric P. [National Oceanography Centre, University of Southampton, European Way, Southampton SO14 3ZH (United Kingdom); Nabi, Abdul [Department of Chemistry, University of Balochistan, Quetta (Pakistan); Worsfold, Paul J., E-mail: pworsfold@plymouth.ac.uk [School of Earth, Ocean and Environmental Sciences (SEOES), University of Plymouth, Drake Circus, Plymouth PL4 8AA (United Kingdom)

    2009-10-12

    A detailed investigation into the performance of two flow injection-chemiluminescence (FI-CL) manifolds (with and without a preconcentration column) for the determination of sub-nanomolar dissolved iron (Fe(II) + Fe(III)), following the reduction of Fe(III) by sulphite, in seawater is described. Kinetic experiments were conducted to examine the efficiency of reduction of inorganic Fe(III) with sulphite under different conditions and a rigorous study of the potential interference caused by other transition metals present in seawater was conducted. Using 100 {mu}M concentrations of sulphite a reduction time of 4 h was sufficient to quantitatively reduce Fe(III) in seawater. Under optimal conditions, cobalt(II) and vanadium(IV)/(III) were the major positive interferences and strategies for their removal are reported. Specifically, cobalt(II) was masked by the addition of dimethylglyoxime to the luminol solution and vanadium(IV) was removed by passing the sample through an 8-hydroxyquinoline column in a low pH carrier stream. Manganese(II) also interfered by suppression of the CL response but this was not significant at typical open ocean concentrations.

  16. Flow-based determination of methionine in pharmaceutical formulations exploiting TGA-capped CdTe quantum dots for enhancing the luminol-KIO{sub 4} chemiluminescence

    Energy Technology Data Exchange (ETDEWEB)

    Zhou, Min, E-mail: mzhou8367@sina.com [Key Laboratory of Eco-Environment-Related Polymer Materials, Ministry of Education, Key Laboratory of Polymer Materials of Gansu Province, Key Laboratory of Bioelectrochemistry & Environmental Analysis of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070 (China); Wang, Ailian [Key Laboratory of Eco-Environment-Related Polymer Materials, Ministry of Education, Key Laboratory of Polymer Materials of Gansu Province, Key Laboratory of Bioelectrochemistry & Environmental Analysis of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070 (China); Jiuquan Enviromental Protection Bureau, Jiuquan 735000 (China); Li, Cong; Luo, Xiaowei; Ma, Yongjun [Key Laboratory of Eco-Environment-Related Polymer Materials, Ministry of Education, Key Laboratory of Polymer Materials of Gansu Province, Key Laboratory of Bioelectrochemistry & Environmental Analysis of Gansu Province, College of Chemistry and Chemical Engineering, Northwest Normal University, Lanzhou 730070 (China)

    2017-03-15

    A novel flow-injection chemiluminescence method (FI-CL) was established for the determination of methionine in this paper, based on its strong enhancement on CL intensity of the luminol-KIO{sub 4} system catalyzed by thioglycolic acid-capped CdTe quantum dots in alkaline media. Under the optimized conditions, the relative CL intensity was in proportion to methionine concentration in the range from 1.0×10{sup −8} to 1.0×10{sup −5} g mL{sup −1} with a detection limit of 6.6×10{sup −9} g mL{sup −1} (3σ). The relative standard deviation (RSD) of the CL intensity for 1.0×10{sup −6} g mL{sup −1} standard methionine solution was 0.97% (n=11). The proposed method was successfully applied to determine methionine in commercial pharmaceutical formulations with recoveries between 98.0% and 101.9%. The possible CL mechanism was discussed as well. - Graphical abstract: Methionine in commercial pharmaceutical formulations was determined by flow-injection chemiluminescence and the possible chemiluminescence mechanism was discussed as well.

  17. Highly sensitive trivalent copper chelate–luminol chemiluminescence system for capillary electrophoresis chiral separation and determination of ofloxacin enantiomers in urine samples

    Directory of Open Access Journals (Sweden)

    Hao-Yue Xie

    2014-12-01

    Full Text Available A simple, fast and sensitive capillary electrophoresis (CE strategy combined with chemiluminescence (CL detection for analysis of ofloxacin (OF enantiomers was established in the present work. Sulfonated β-cyclodextrin (β-CD was used as the chiral additive being added into the running buffer of luminol–diperiodatocuprate (III (K5[Cu(HIO62], DPC chemiluminescence system. Under the optimum conditions, the proposed method was successfully applied to separation and analysis of OF enantiomers with the detection limits (S/N=3 of 8.0 nM and 7.0 nM for levofloxacin and dextrofloxacin, respectively. The linear ranges were both 0.010–100 μM. The method was utilized for analyzing OF in urine; the results obtained were satisfactory and recoveries were 89.5–110.8%, which demonstrated the reliability of this method. This approach can also be further extended to analyze different commercial OF medicines. Keywords: Ofloxacin, Chiral analysis, Sulfonated-β-CD, Capillary electrophoresis, Chemiluminescence

  18. Development of a reagentless electrochemiluminescent electrode for flow injection analysis using copolymerised luminol/aniline on nano-TiO2 functionalised indium-tin oxide glass.

    Science.gov (United States)

    Liu, Chao; Wei, Xiuhua; Tu, Yifeng

    2013-07-15

    In this study, a nano-structured copolymer of luminol/aniline (PLA) was deposited onto nano-TiO2-functionalised indium tin oxide (ITO)-coated glass by electrochemical polymerisation using cyclic voltammetry (CV). The resulting reagentless electrochemiluminescent (ECL) electrode (ECLode) can be used for flow injection analysis (FIA). The properties of the ECLode were characterised by CV, electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). The ECLode has high background ECL emission as well as excellent stability and reproducibility, and yielding sensitive response towards target analytes. The ECL emissions of the ECLode were 50 times higher than PLA/ITO, and 500 times higher than polyluminol (PL)/ITO. The ECLode showed sensitive responses to reactive oxygen species (ROSs), permitting its application for determination of antioxidants by quenching. Under optimised conditions, an absolute detection limit of 69.9 pg was obtained for resveratrol, comparable to the highest levels of sensitivity achieved by other methods. Thus, the gross antioxidant content of red wine was determined, with satisfactory recoveries between 87.6% and 108.3%. These results suggest a bright future for the use of the ECLode for single-channel FIA due to its high sensitivity, accuracy and reproducibility. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Fe3O4 and metal-organic framework MIL-101(Fe) composites catalyze luminol chemiluminescence for sensitively sensing hydrogen peroxide and glucose.

    Science.gov (United States)

    Qian Tang, Xue; Dan Zhang, Yi; Wei Jiang, Zhong; Mei Wang, Dong; Zhi Huang, Cheng; Fang Li, Yuan

    2018-03-01

    In this work, Fe 3 O 4 and metal-organic framework MIL-101(Fe) composites (Fe 3 O 4 /MIL-101(Fe)) was demonstrated to possess excellent catalytic property to directly catalyze luminol chemiluminescence without extra oxidants. We utilized Fe 3 O 4 /MIL-101(Fe) to develop a ultra-sensitive quantitative analytical method for H 2 O 2 and glucose. The possible mechanism of the chemiluminescence reaction had been investigated. Under optimal conditions, the relative chemiluminescence intensity was linearly proportional to the logarithm of H 2 O 2 concentration in the range of 5-150nM with a limit of detection of 3.7nM (signal-to-noise ratio = 3), and glucose could be linearly detected in the range from 5 to 100nM and the detection limit was 4.9nM (signal-to-noise ratio = 3). Furthermore, the present approach was successfully applied to quantitative determination of H 2 O 2 in medical disinfectant and glucose in human serum samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. A sensitive inhibition chemiluminescence method for the determination of 6-mercaptopurine in tablet and biological fluid using the reaction of luminol-Ag(III) complex in alkaline medium

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Hanwen, E-mail: hanwen@hbu.edu.cn [College of Chemistry and Environmental Science, Hebei University, Key Laboratory of Analytical Science and Technology of Hebei Province, Baoding 071002 (China); Wang, Ting; Liu, Xuyang; Chen, Peiyun [College of Chemistry and Environmental Science, Hebei University, Key Laboratory of Analytical Science and Technology of Hebei Province, Baoding 071002 (China)

    2013-02-15

    A sensitive inhibition chemiluminescence (CL) method for the determination of 6-mercaptopurine (6-MP) is developed. The mechanism of the CL reaction between Ag(III) complex {l_brace}[Ag(HIO{sub 6}){sub 2}]{sup 5-}{r_brace} and luminol in alkaline solution was proposed, along with the inhibition mechanism of 6-MP on the CL emission. The inhibition degree of CL emission was proportional to the logarithm of 6-MP concentration. The effects of the reaction conditions on CL emission and inhibition were examined. Under the optimized conditions, the detection limit (s/n=3) was 3.7 Multiplication-Sign 10{sup -10} g ml{sup -1}. The recoveries of 6-MP were in the range of 97.7-105% with the RSD of 2.1-3.4% (n=5) for tablet samples, 103-106% with the RSDs of 1.1-2.1% for spiked serum sample, and 97.2-101% with the RSD of 2.0-4.5% for spiked urine sample. The accuracy of this method for the tablet analysis was examined by comparing with the pharmacopoeia method. The proposed method was used for the determination of 6-MP at clinically relevant concentrations in real urine and serum samples with satisfactory results. - Highlights: Black-Right-Pointing-Pointer A sensitive inhibition chemiluminescence (CL) method for the determination of 6-MP is developed. Black-Right-Pointing-Pointer The inhibition mechanism of 6-MP on the CL emission was proposed. Black-Right-Pointing-Pointer The detection limit was 3.7 Multiplication-Sign 10{sup -10} g ml{sup -1}. Black-Right-Pointing-Pointer The accuracy was examined by comparing with the pharmacopoeia method.

  1. Determination of phenols by flow injection and liquid chromatography with on-line quinine-sensitized photo-oxidation and quenched luminol chemiluminescence detection

    International Nuclear Information System (INIS)

    Zhang Wei; Danielson, Neil D.

    2003-01-01

    An on-line quinine-sensitized photo-oxidation with quenched chemiluminescence (CL) detection method is developed for phenols using flow injection (FI) and liquid chromatography (LC). This detection method is based on the decrease of light emission from the luminol CL reaction due to the photo-oxidation of phenols that scavenge the photogenerated reactive oxygen species (e.g. singlet oxygen ( 1 O 2 ) and superoxide (O 2 · - )). On-line photo-oxidation is achieved using a coil photo-reactor made from fluoroethylene-propylene copolymer tubing (3048 mmx0.25 mm i.d.) coiled around a mercury UV lamp. A buffer of pH 7 and a concentration of 350 μM for quinine sulfate are determined optimum for the sensitized photo-oxidation. Using a carrier system flow rate of 60 μl/min, calibration curves taken by FI for 10 phenolic compounds in aqueous solutions showed this decreasing sensitivity order: 4-chlorophenol, phenol, 4-nitrophenol, 3-hydroxy-L-kynurenine, 2-nitrophenol, salicylate, 3-nitrophenol, catechol, 2,4-dinitrophenol, and 2,4-dichlorophenol. This detection method using two tandem coil photo-reactors is also applied for the LC separation of phenol, 4-nitrophenol and 4-chlorophenol on an octadecyl (C18) silica LC column using acetonitrile-H 2 O (40:60, v/v) as a mobile phase. The quenched CL detection limits (about 1 μM or 20 pmol) for phenol and 4-chlorophenol are comparable to those for UV detection at 254 nm. Some selectivity in the quenched CL detection is evident by no interference in the FI phenol response even when benzaldehyde and phenethanol concentrations are 8 and 15 times that of phenol

  2. Comparative study of β-glucan induced respiratory burst measured by nitroblue tetrazolium assay and real-time luminol-enhanced chemiluminescence assay in common carp (Cyprinus carpio L.).

    Science.gov (United States)

    Vera-Jimenez, N I; Pietretti, D; Wiegertjes, G F; Nielsen, M E

    2013-05-01

    The respiratory burst is an important feature of the immune system. The increase in cellular oxygen uptake that marks the initiation of the respiratory burst is followed by the production of reactive oxygen species (ROS) such as superoxide anion and hydrogen peroxide which plays a role in the clearance of pathogens and tissue regeneration processes. Therefore, the respiratory burst and associated ROS constitute important indicators of fish health status. This paper compares two methods for quantitation of ROS produced during the respiratory burst in common carp: the widely used, single-point measurement based on the intracellular reduction of nitroblue tetrazolium (NBT) and a real-time luminol-enhanced assay based on the detection of native chemiluminescence. Both assays allowed for detection of dose-dependent changes in magnitude of the respiratory burst response induced by β-glucans in head kidney cells of carp. However, whereas the NBT assay was shown to detect the production of only superoxide anions, the real-time luminol-enhanced assay could detect the production of both superoxide anions and hydrogen peroxide. Only the chemiluminescence assay could reliably record the production of ROS on a real-time scale at frequent and continual time intervals for time course experiments, providing more detailed information on the respiratory burst response. The real-time chemiluminescence assay was used to measure respiratory burst activity in macrophage and neutrophilic granulocyte-enriched head kidney cell fractions and total head kidney cell suspensions and proved to be a fast, reliable, automated multiwell microplate assay to quantitate fish health status modulated by β-glucans. Copyright © 2013 Elsevier Ltd. All rights reserved.

  3. A highly sensitive NADH sensor based on a mycelium-like nanocomposite using graphene oxide and multi-walled carbon nanotubes to co-immobilize poly(luminol) and poly(neutral red) hybrid films.

    Science.gov (United States)

    Chiang Lin, Kuo; Yu Lai, Szu; Ming Chen, Shen

    2014-08-21

    Hybridization of poly(luminol) (PLM) and poly(neutral red) (PNR) has been successfully performed and further enhanced by a conductive and steric hybrid nanotemplate using graphene oxide (GO) and multi-walled carbon nanotubes (MWCNTs). The morphology of the PLM-PNR-MWCNT-GO mycelium-like nanocomposite is studied by SEM and AFM and it is found to be electroactive, pH-dependent, and stable in the electrochemical system. It shows electrocatalytic activity towards NADH with a high current response and low overpotential. Using amperometry, it has been shown to have a high sensitivity of 288.9 μA mM(-1) cm(-2) to NADH (Eapp. = +0.1 V). Linearity is estimated in a concentration range of 1.33 × 10(-8) to 1.95 × 10(-4) M with a detection limit of 1.33 × 10(-8) M (S/N = 3). Particularly, it also shows another linear range of 2.08 × 10(-4) to 5.81 × 10(-4) M with a sensitivity of 151.3 μA mM(-1) cm(-2). The hybridization and activity of PLM and PNR can be effectively enhanced by MWCNTs and GO, resulting in an active hybrid nanocomposite for determination of NADH.

  4. A novel "dual-potential" electrochemiluminescence aptasensor array using CdS quantum dots and luminol-gold nanoparticles as labels for simultaneous detection of malachite green and chloramphenicol.

    Science.gov (United States)

    Feng, Xiaobin; Gan, Ning; Zhang, Huairong; Yan, Qing; Li, Tianhua; Cao, Yuting; Hu, Futao; Yu, Hongwei; Jiang, Qianli

    2015-12-15

    A novel type of "dual-potential" electrochemiluminescence (ECL) aptasensor array was fabricated on a homemade screen-printed carbon electrode (SPCE) for simultaneous detection of malachite green (MG) and chloramphenicol (CAP) in one single assay. The SPCE substrate consisted of a common Ag/AgCl reference electrode, carbon counter electrode and two carbon working electrodes (WE1 and WE2). In the system, CdS quantum dots (QDs) were modified on WE1 as cathode ECL emitters and luminol-gold nanoparticles (L-Au NPs) were modified on WE2 as anode ECL emitters. Then the MG aptamer complementary strand (MG cDNA) and CAP aptamer complementary strand (CAP cDNA) were attached on CdS QDs and L-Au NPs, respectively. The cDNA would hybridize with corresponding aptamer that was respectively tagged with cyanine dye (Cy5) (as quenchers of CdS QDs) and chlorogenic acid (CA) (as quenchers of l-Au NPs) using poly(ethylenimine) (PEI) as a bridging agent. PEI could lead to a large number of quenchers on the aptamer, which increased the quenching efficiency. Upon MG and CAP adding, the targets could induce strand release due to the highly affinity of analytes toward aptamers. Meanwhile, it could release the Cy5 and CA, which recovered cathode ECL of CdS QDs and anode ECL of L-Au NPs simultaneously. This "dual-potential" ECL strategy could be used to detect MG and CAP with the linear ranges of 0.1-100 nM and 0.2-150 nM, with detection limits of 0.03 nM and 0.07 nM (at 3sB), respectively. More importantly, this designed method was successfully applied to determine MG and CAP in real fish samples and held great potential in the food analysis. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Luminol-and lucigenin-amplified chemiluminescence with rat liver microsomes. Kinetics and influence of ascorbic acid, glutathione, dimethylsulfoxide, N-t-butyl-a-phenyl-nitrone, copper-ions and a copper complex, catalase, superoxide dismutase, hexobarbital and aniline.

    Science.gov (United States)

    Klinger, W; Karge, E; Kretzschmar, M; Rost, M; Schulze, H P; Dargel, R; Reinemann, C; Rein, H

    1996-07-01

    For the investigation of luminol (LM)-and lucigenin (LC)-amplified chemiluminescence (CL) in rat liver microsomes using both a liquid-scintillation counter (LKB/Wallac 1219 Rackbeta) and a Berthold luminometer (AutoLumat LB 953) optimal incubation mixtures and conditions and basic kinetics have been established. Whereas calibration curves for both LM- and LC-CL are performed with hydrogenperoxide (LC quantum yield is 6.25 fold higher as that of LM), distinct differences were revealed with microsomes, indicating that different reactive oxygen species (ROS) are determined: Both LM- and LC-CL follow the kinetics of enzymatic reactions in terms of dependence on protein and NADPH or NADH concentration, time course, temperature etc., but with differences. LM-CL does not work without addition of Fe2+, whereas LC-CL does. Both copper ions and copper bound in a complex abolish CL, LC-CL being much more sensitive. Isolated cytochrome P-450 (P450) and NADPH P450 reductase from liver of pheno-barbital treated rats alone proved to be inactive in LM-and LC-CL production, whereas te combination 1:1 without and with addition of lipid was highly active in both LM-and LC-CL. Ascorbic acid and glutathione as scavengers diminish both LM- and LC-CL in concentrations higher then 10(5). Dimethyl-sulfoxide (DMSO) was ineffective in LM-CL up to concentrations of 0.2 M, the very high concentration of 2 M diminished LM-CL only to 1/3. LC-CL was diminished starting at concentrations of 100 mM and at 2 M only 10% of maximum LC-CL was observed. The trap substance N-t-butyl-a-phenylnitrone (BNP) also diminished LC-CL more effectively than LM-CL. Clearcut differences were revealed by the addition of catalase and superoxide dismutase: both enzymes diminished LM-CL only, without any influence on LC-CL. Hexobarbital, a potent uncoupler of P450, enhances LM-CL fivefold, whereas LC-CL is barely influenced. Aniline (without uncoupling capability) decreased both LM-and LC-CL increasingly with increasing

  6. Monosodium Luminol for Improving Brain Function in Gulf War Illness

    Science.gov (United States)

    2016-10-01

    much attention. First, veterans who were stationed in the battlefield areas believed to have consumed pills of pyridostigmine bromide (PB) during...966, 2007; Yassa and Stark, Trends Neurosci , 34: 515-525). Maintenance of this function depends upon the integrity of the hippocampus particularly...as described in our published report (Hattiangady et al., Frontiers in Behavioral Neuroscience , 2014). We examined rats belonging to different

  7. Monosodium Luminol for Improving Brain Function in Gulf War Illness

    Science.gov (United States)

    2015-10-01

    of BrdU+ cells (i.e. newly born cells), doublecortin (DCX, a marker of newly born neurons ), glial fibrillary acidic protein, (GFAP, a marker of...can reduce hydrogen peroxide and short chain organic, fatty acid , and phospholipid hydroperoxides. It is also believed to play a role in the...gene encoding thioredoxin reductase 1, which reduces thioredoxins as well as other substrates, and plays a role in selenium metabolism and protection

  8. Differential stimulation of luminol-enhanced chemiluminescence (CL) and arachidonic acid metabolism in rat peritoneal neutrophils

    Energy Technology Data Exchange (ETDEWEB)

    Sturm, R.J.; Adams, L.M.; Cullinan, C.A.; Berkenkopf, J.W.; Weichman, B.M.

    1986-03-05

    Phorbol 12-myristate, 13-acetate (PMA) induced the production of radical oxygen species (ROS) from rat peritoneal neutrophils as assessed by CL. ROS generation occurred in a time- (maximum at 13.5 min) and dose- (concentration range of 1.7-498 nM) related fashion. However, 166 nM PMA did not induce either cyclooxygenase (CO) or lipoxygenase (LPO) product formation by 20 min post-stimulation. Conversely, A23187, at concentrations between 0.1 and 10 ..mu..M, stimulated both pathways of arachidonic acid metabolism, but had little or no effect upon ROS production. When suboptimal concentrations of PMA (5.5 nM) and A23187 (0.1-1 ..mu..M) were coincubated with the neutrophils, a synergistic ROS response was elicited. However, arachidonic acid metabolism in the presence of PMA was unchanged relative to A12187 alone. Nordihydroguaiaretic acid (NDGA) inhibited both PMA-induced CL (IC/sub 50/ = 0.9 ..mu..M) and A23187-induced arachidonic acid metabolism (IC/sub 50/ = 1.7 ..mu..M and 6.0 ..mu..M for LPO and CO, respectively). The mixed LPO-CO inhibitor, BW755C, behaved in a qualitatively similar manner to NDGA, whereas the CO inhibitors, indomethacin, piroxicam and naproxen had no inhibitory effect on ROS generation at concentrations as high as 100 ..mu..M. These results suggest that NDGA and BW755C may inhibit CL and arachidonic acid metabolism by distinct mechanisms in rat neutrophils.

  9. Liquid chromatography with luminol-based electrochemiluminescence detection determination of histamine

    NARCIS (Netherlands)

    Steijger, O.M.; Kamminga, D.A.; Brummelhuis, A.; Lingeman, H.

    1998-01-01

    The liquid chromatographic determination of histamine was achieved by precolumn derivatization with N-(4-aminobutyl)-N-ethylisoluminol isothiocyanate and electrochemiluminescence detection. Detection was carried out using postcolumn on-line electrochemical reagent generation and oxidation of the

  10. High Catalysis Activity of Cu2O Microcrystals to the Electrochemiluminesence of Luminol and H2O2

    DEFF Research Database (Denmark)

    Zhang, Ling; Xu, Guobao; Zhang, Jingdong

    Cuprous oxide (Cu2O) is a classical p-type semiconductor with a direct band gap of 2.17 eV, which is wildly used for solar energy conversion, CO oxidation, and photo catalytic water splitting for the low cost and environmental friendliness. For the energy band positions are favorable to the hydro......Cuprous oxide (Cu2O) is a classical p-type semiconductor with a direct band gap of 2.17 eV, which is wildly used for solar energy conversion, CO oxidation, and photo catalytic water splitting for the low cost and environmental friendliness. For the energy band positions are favorable...... to the hydrogen evolution and oxygen evolution potentials, Cu2O materials also catalyze the reduction of hydrogen peroxide (H2O2), which is an critical molecule in the bodies’ metabolism processes or the industrial catalysis reactions. To improve detection sensitivity of H2O2, people have composed Cu2O materials...... that Cu2O semiconductor microcrystal possess the good catalytic performance to this ECL reaction, which is important to develop the high-efficient and lowcost biosensors....

  11. Encapsulation of Hemin in Metal-Organic Frameworks for Catalyzing the Chemiluminescence Reaction of the H2O2-Luminol System and Detecting Glucose in the Neutral Condition.

    Science.gov (United States)

    Luo, Fenqiang; Lin, Yaolin; Zheng, Liyan; Lin, Xiaomei; Chi, Yuwu

    2015-06-03

    Novel metal-organic frameworks (MOFs) based solid catalysts have been synthesized by encapsulating Hemin into the HKUST-1 MOF materials. These have been first applied in the chemiluminescence field with outstanding performance. The functionalized MOFs not only maintain an excellent catalytic activity inheriting from Hemin but also can be cyclically utilized as solid mimic peroxidases in the neutral condition. The synthesized Hemin@HKUST-1 composites have been used to develop practical sensors for H2O2 and glucose with wide response ranges and low detection limits. It was envisioned that catalyst-functionalized MOFs for chemiluminescence sensing would have promising applications in green, selective, and sensitive detection of target analytes in the future.

  12. Comparative study of B-glucan induced respiratory burst measured by nitroblue tetrazolium assay and real-time luminol-enhanced chemiluminescence assay in common carp (Cyrpinus carpio L.)

    NARCIS (Netherlands)

    Vera-Jimenez, N.I.; Pietretti, D.; Wiegertjes, G.F.; Nielsen, M.E.

    2013-01-01

    The respiratory burst is an important feature of the immune system. The increase in cellular oxygen uptake that marks the initiation of the respiratory burst is followed by the production of reactive oxygen species (ROS) such as superoxide anion and hydrogen peroxide which plays a role in the

  13. 鲁米诺-[铁氰化钾-亚铁氰化钾]-酚磺乙胺化学发光体系%Chemiluminescence Reaction of Luminol-[Potassium Ferric yanide-Potassium Ferrocyanide]-Ethamsylate System

    Institute of Scientific and Technical Information of China (English)

    李银环; 杜建修; 吕九如

    2002-01-01

    实验发现,酚磺乙胺与鲁米诺在碱性溶液中可产生弱的化学发光,铁氰化钾和亚铁氰化钾的存在对该化学发光反应具有很强的催化作用.基于此,建立了基于鲁米诺-铁氰化钾/亚铁氰化钾-酚磺乙胺体系测定酚磺乙胺的流动注射化学发光新方法.酚磺乙胺浓度在4 .0×10-6~1.0×10-4 g/L范围内与化学发光强度具有线性关系.该方法的检出限为1×10-6 g/L,相对标准偏差为1.4%(2.0×10-5 g/L 酚磺乙胺溶液,n=11).该方法用于止血敏注射液中酚磺乙胺的含量测定,结果令人满意.同时 ,还对可能的化学发光反应机理进行了初步探讨.

  14. Bulletin of Materials Science | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    Recent analytical applications of nanoparticle sensitized lucigenin and luminol ..... Analysis on insulator–metal transition in yttrium doped LSMO from electron ... Synthesis and luminescence properties of Eu-activated Ca4Mg5(PO4)6 for ...

  15. Determination of monoamine neurotransmitters and their metabolites in a mouse brain microdialysate by coupling high-performance liquid chromatography with gold nanoparticle-initiated chemiluminescence

    International Nuclear Information System (INIS)

    Li Na; Guo Jizhao; Liu Bo; Yu Yuqi; Cui Hua; Mao Lanqun; Lin Yuqing

    2009-01-01

    Our previous work showed that gold nanoparticles could trigger chemiluminescence (CL) between luminol and AgNO 3 . In the present work, the effect of some biologically important reductive compounds, including monoamine neurotransmitters and their metabolites, reductive amino acids, ascorbic acid, uric acid, and glutathione, on the novel CL reaction were investigated for analytical purpose. It was found that all of them could inhibit the CL from the luminol-AgNO 3 -Au colloid system. Among them, monoamine neurotransmitters and their metabolites exhibited strong inhibition effect. Taking dopamine as a model compound, the CL mechanism was studied by measuring absorption spectra during the CL reaction and the reaction kinetics via stopped-flow technique. The CL inhibition mechanism is proposed to be due to that these tested compounds competed with luminol for AgNO 3 to inhibit the formation of luminol radicals and to accelerate deposition of Ag atoms on surface of gold nanoparticles, leading to a decrease in CL intensity. Based on the inhibited CL, a novel method for simultaneous determination of monoamine neurotransmitters and their metabolites was developed by coupling high-performance liquid chromatography with this CL reaction. The new method was successfully applied to determine the compounds in a mouse brain microdialysate. Compared with the reported HPLC-CL methods, the proposed method is simple, fast, and could determine more analytes. Moreover, the limits of linear ranges for NE, E, and DA using the proposed method were one order of magnitude lower than the luminol system without gold nanoparticles.

  16. Effect of the dimetilsulfoxido in the response chemiluminescent and the consumption of oxygen of neutrophils activated human

    International Nuclear Information System (INIS)

    Garcia, J.

    2001-01-01

    Dimethylsulfoxide (DMSO), a hydroxyl radical scavenger, exerted a dose dependent inhibition on the luminol and lucigenin-enhanced chemiluminescent responses of human neutrophils activated with soluble and particulate stimulants. DMSO inhibition of the luminol chemiluminescense induced by calcium ionophore A23187 was probably due to OH scavenging, whereas inhibition of the lucigenin chemiluminescence suggested DMSO negatively affects the NADPH-dependent membrane oxidase of neutrophils. In agreement with this, DMSO moderately inhibited O2 consumption in PMN suspensions stimulated with chemotactic peptide and opsonized zymosan-induced luminol chemiluminescense was observed only when added before or in conjunction with stimulants, whereas A23187-induced chemiluminescense was inhibited by DMSO regardless of time of addition. Washing of DMSO-treated PMN resulted in increased luminol enhanced chemiluminescense in response to chemotactic peptide and opsonized zymosan. This is consistent with the idea that DMSO may be interfering with activation of the membrane subunits of the oxidase by translocation and docking of the cytoplasmic, regulatory subunits. These data imply that DMSO inhibits neutrophil chemiluminescense both by OH scavenging and interfering with oxidase activation. Key words:Dimethylsulfoxide, chemiluminescent, luminol, lucigenin,neutrophils [es

  17. [Development and Application of Catalytic Tyrosine Modification].

    Science.gov (United States)

    Sato, Shinichi; Tsushima, Michihiko; Nakamura, Kosuke; Nakamura, Hiroyuki

    2018-01-01

     The chemical labeling of proteins with synthetic probes is a key technique used in chemical biology, protein-based therapy, and material science. Much of the chemical labeling of native proteins, however, depends on the labeling of lysine and cysteine residues. While those methods have significantly contributed to native protein labeling, alternative methods that can modify different amino acid residues are still required. Herein we report the development of a novel methodology of tyrosine labeling, inspired by the luminol chemiluminescence reaction. Tyrosine residues are often exposed on a protein's surface and are thus expected to be good targets for protein functionalization. In our studies so far, we have found that 1) hemin oxidatively activates luminol derivatives as a catalyst, 2) N-methyl luminol derivative specifically forms a covalent bond with a tyrosine residue among the 20 kinds of natural amino acid residues, and 3) the efficiency of tyrosine labeling with N-methyl luminol derivative is markedly improved by using horseradish peroxidase (HRP) as a catalyst. We were able to use molecular oxygen as an oxidant under HRP/NADH conditions. By using these methods, the functionalization of purified proteins was carried out. Because N-methyl luminol derivative is an excellent protein labeling reagent that responds to the activation of peroxidase, this new method is expected to open doors to such biological applications as the signal amplification of HRP-conjugated antibodies and the detection of protein association in combination with peroxidase-tag technology.

  18. Applications of chemiluminescence to bacterial analysis

    Science.gov (United States)

    Searle, N. D.

    1975-01-01

    Luminol chemiluminescence method for detecting bacteria was based on microbial activation of the oxidation of the luminol monoanion by hydrogen peroxide. Elimination of the prior lysing step, previously used in the chemiluminescence technique, was shown to improve considerably the reproducibility and accuracy of the method in addition to simplifying it. An inexpensive, portable photomultiplier detector was used to measure the maximum light intensity produced when the sample is added to the reagent. Studies of cooling tower water show that the luminol chemiluminescence technique can be used to monitor changes in viable cell population both under normal conditions and during chlorine treatment. Good correlation between chemiluminescence and plate counts was also obtained in the analysis of process water used in paper mills. This method showed good potential for monitoring the viable bacteria populations in activated sludge used in waste treatment plants to digest organic matter.

  19. Study on the luminescence behavior of sulfobutylether-β-cyclodextrin with risperidone and its analytical application.

    Science.gov (United States)

    Wu, Min; Chen, Donghua; Song, Zhenghua

    2012-10-01

    The interaction of sulfobutylether-β-cyclodextrin (SBE-β-CD) with risperidone (RISP) was first described with luminol-SBE-β-CD chemiluminescence (CL) system by flow injection analysis (FIA). In luminol-SBE-β-CD CL system, the 1:1 SBE-β-CD···luminol(*) complexation could enhance CL intensity of luminol and produce the effect of complexation enhancement of CL (CEC). It was found that RISP could quench the CL intensity of SBE-β-CD···luminol(*) and caused the effect of complexation enhancement of quenching (CEQ), the formation constant K(R-CD) 3.4×10(4) L mol(-1) and the stoichiometric ratio 1:1 of RISP···SBE-β-CD complex were obtained by the proposed CL model. Association degree α 0.036 of RISP···SBE-β-CD complex was also given by CL method. Based on the linear relationship to the decrement of luminol-SBE-β-CD-RISP CL intensity and the logarithm of RISP concentration, RISP also can be quantified in the linear range of 3.0-500.0 nmol L(-1) with a detection limit of 1.0 nmol L(-1) (3σ). The proposed method was successfully applied to monitoring excreted RISP in human urine. It was found that RISP reached its maximum after oral administration for 1.5 h with the total excretion of 14.26% within 8.5 h; the elimination rate constant k and half-life time t(1/2) were 0.474 and 1.5 h, respectively. Copyright © 2012 Elsevier B.V. All rights reserved.

  20. Lyoluminescence characteristics of trehalose dihydrate

    Energy Technology Data Exchange (ETDEWEB)

    Raman, Anand E-mail: anandr@apsara.barc.ernet.in; Oommen, Issac K.; Sharma, D.N

    2001-03-01

    Lyoluminescence (LL) of trehalose dihydrate showed a linear gamma ray sensitivity in the dose range 0.1-5x10{sup 2} Gy. At increased sample weight dissolved, the LL/mg showed a reduction when the microcontroller based integrated measurement system was used. The LL/mg was found to be independent of the sample weight when the instantaneous peak output observed by the electrometer amplifier on dissolution of the phosphor was taken as a measure of LL yield. The LL spectral measurement of trehalose dihydrate in luminol solution (LL sensitizer) confirms an energy transfer from the radiation induced free radicals to luminol molecule to produce light.

  1. Lyoluminescence characteristics of trehalose dihydrate

    International Nuclear Information System (INIS)

    Raman, Anand; Oommen, Issac K.; Sharma, D.N.

    2001-01-01

    Lyoluminescence (LL) of trehalose dihydrate showed a linear gamma ray sensitivity in the dose range 0.1-5x10 2 Gy. At increased sample weight dissolved, the LL/mg showed a reduction when the microcontroller based integrated measurement system was used. The LL/mg was found to be independent of the sample weight when the instantaneous peak output observed by the electrometer amplifier on dissolution of the phosphor was taken as a measure of LL yield. The LL spectral measurement of trehalose dihydrate in luminol solution (LL sensitizer) confirms an energy transfer from the radiation induced free radicals to luminol molecule to produce light

  2. Fiber-Optic Chemiluminescent Biosensors for Monitoring Aqueous Alcohols and Other Water Quality Parameters

    Science.gov (United States)

    Verostko, Charles E. (Inventor); Atwater, James E. (Inventor); Akse, James R. (Inventor); DeHart, Jeffrey L. (Inventor); Wheeler, Richard R. (Inventor)

    1998-01-01

    A "reagentless" chemiluminescent biosensor and method for the determination of hydrogen peroxide, ethanol and D-glucose in water is disclosed. An aqueous stream is basified by passing it through a solid phase base bed. Luminol is then dissolved in the basified effluent at a controlled rate. Oxidation of the luminol is catalyzed by the target chemical to produce emitted light. The intensity of the emitted light is detected as a measure of the target chemical concentration in the aqueous stream. The emitted light can be transmitted by a fiber optic bundle to a remote location from the aqueous stream for a remote reading of the target chemical concentration.

  3. New electrochemiluminescence catalyst: Cu2O semiconductor crystal and the enhanced activity of octahedra synthesized by iodide ions coordination

    DEFF Research Database (Denmark)

    Zhang, Ling; Qi, Liming; Gao, Wenyue

    2017-01-01

    Cuprous oxide (Cu2O) crystals are usually used as the photocatalysts of water splittings and carbon monoxide oxidations. Herein, we report the new catalytic properties of Cu2O to the electrochemilumenescence (ECL) reactions of luminol and oxygen. Adjusting the shape of Cu2O microcrystals from cub...

  4. Additional information to the in vitro antioxidant activity of Ginkgo biloba L

    NARCIS (Netherlands)

    Lugasi, A; Horvahovich, P; Dworschák, E

    The in vitro antioxidant and free radical scavenging activity of the ethanol extract from Ginkgo biloba L. was examined in different systems. The extract showed hydrogen-donating ability, reducing power, copper-binding property, free radical scavenging activity in a H2O2/.OH-luminol system and it

  5. Determination of hydrogen peroxide in water by chemiluminescence detection, (1). Flow injection type hydrogen peroxide detection system

    International Nuclear Information System (INIS)

    Yamashiro, Naoya; Uchida, Shunsuke; Satoh, Yoshiyuki; Morishima, Yusuke; Yokoyama, Hiroaki; Satoh, Tomonori; Sugama, Junichi; Yamada, Rie

    2004-01-01

    A flow injection type hydrogen peroxide detection system with a sub-ppb detection limit has been developed to determine hydrogen peroxide concentration in water sampled from a high temperature, high pressure hydrogen peroxide water loop. The hydrogen peroxide detector is based on luminol chemiluminescence spectroscopy. A small amount of sample water (20 μl) is mixed with a reagent mixture, an aqueous solution of luminol and Co 2+ catalyst, in a mixing cell which is installed just upstream from the detection cell. The optimum values for pH and the concentrations of luminol and Co 2+ ion have been determined to ensure a lower detectable limit and a higher reproducibility. The photocurrent detected by the detection system is expressed by a linear function of the hydrogen peroxide concentration in the region of lower concentration ([H 2 O 2 ] 2 O 2 ] in the region of higher concentration ([H 2 O 2 ] > 10 ppb). The luminous intensity of luminol chemiluminescence is the highest when pH of the reagent mixture is 11.0. Optimization of the major parameters gives the lowest detectable limit of 0.3 ppb. (author)

  6. Chemiluminescence measurements as an identification method for gamma-irradiated foodstuffs

    Energy Technology Data Exchange (ETDEWEB)

    Boegl, W; Heide, L

    1985-01-01

    Samples of 19 different spices, milk powder, whole onions and frozen chicken were exposed to a Co-60 source with radiation doses up to 10/sup 4/ Gy. The subsequent reaction of the irradiated foodstuffs in a luminol solution resulted in light emission (chemiluminescence). This effect can be used as an indicator of radiation treatment.

  7. STIMULATION OF OXIDANT PRODUCTION IN ALVEOLAR MACROPHAGES BY POLLUTANT AND LATEX PARTICLES

    Science.gov (United States)

    Air pollutant dusts as well as chemically defined particles were examined for their activating effect on oxidant production (O2- and H2O2) in guinea pig alveolar macrophages (AM). Oxidant production was measured as chemiluminescence of albumin-bound luminol. All particles examine...

  8. Determination of monoamine neurotransmitters and their metabolites in a mouse brain microdialysate by coupling high-performance liquid chromatography with gold nanoparticle-initiated chemiluminescence

    Energy Technology Data Exchange (ETDEWEB)

    Li Na; Guo Jizhao; Liu Bo; Yu Yuqi [Department of Chemistry, University of Science and Technology of China (USTC), JinZhai Road No: 96, 230026 Hefei, Anhui (China); Cui Hua, E-mail: hcui@ustc.edu.cn [Department of Chemistry, University of Science and Technology of China (USTC), JinZhai Road No: 96, 230026 Hefei, Anhui (China); Mao Lanqun; Lin Yuqing [Beijing National Laboratory for Molecular Sciences, Institute of Chemistry, Chinese Academy of Sciences (CAS), 100080 Beijing (China)

    2009-07-10

    Our previous work showed that gold nanoparticles could trigger chemiluminescence (CL) between luminol and AgNO{sub 3}. In the present work, the effect of some biologically important reductive compounds, including monoamine neurotransmitters and their metabolites, reductive amino acids, ascorbic acid, uric acid, and glutathione, on the novel CL reaction were investigated for analytical purpose. It was found that all of them could inhibit the CL from the luminol-AgNO{sub 3}-Au colloid system. Among them, monoamine neurotransmitters and their metabolites exhibited strong inhibition effect. Taking dopamine as a model compound, the CL mechanism was studied by measuring absorption spectra during the CL reaction and the reaction kinetics via stopped-flow technique. The CL inhibition mechanism is proposed to be due to that these tested compounds competed with luminol for AgNO{sub 3} to inhibit the formation of luminol radicals and to accelerate deposition of Ag atoms on surface of gold nanoparticles, leading to a decrease in CL intensity. Based on the inhibited CL, a novel method for simultaneous determination of monoamine neurotransmitters and their metabolites was developed by coupling high-performance liquid chromatography with this CL reaction. The new method was successfully applied to determine the compounds in a mouse brain microdialysate. Compared with the reported HPLC-CL methods, the proposed method is simple, fast, and could determine more analytes. Moreover, the limits of linear ranges for NE, E, and DA using the proposed method were one order of magnitude lower than the luminol system without gold nanoparticles.

  9. MIL-53(Fe) MOF-mediated catalytic chemiluminescence for sensitive detection of glucose.

    Science.gov (United States)

    Yi, Xueling; Dong, Wenfei; Zhang, Xiaodan; Xie, Jianxin; Huang, Yuming

    2016-12-01

    Various analytical applications of metal-organic frameworks (MOFs) have been rapidly developed in the past few years. However, the employment of MOFs as catalysts in chemiluminescence (CL) analysis is rare. Here, for the first time, we found that MIL-53(Fe) MOFs could significantly enhance the CL of luminol in the presence of H 2 O 2 in an alkaline medium. The CL intensity in the luminol-H 2 O 2 -MIL-53(Fe) system was about 20 times higher than that in the luminol-H 2 O 2 system. Moreover, the XRD pattern of MIL-53(Fe) after CL reaction was almost the same as that of the original MIL-53(Fe), confirming the catalytic role of MIL-53(Fe) in the luminol-H 2 O 2 -MIL-53(Fe) system. The possible mechanism behind the enhancing phenomenon was discussed based on the results from the CL spectra, FL probe experiments, and active oxygen species measurements. By coupling with the glucose oxidase-based catalytic oxidation reaction, a sensitive and selective CL method was developed for the detection of glucose. There is a linear relationship between the logarithm of CL intensity and the logarithm of glucose concentration in the range from 0.1 to 10 μM, and a detection limit of 0.05 μM (S/N = 3) is obtained. The proposed method has been applied to the determination of glucose in human serum samples with satisfactory results. Graphical abstract MIL-53(Fe) MOFs are found to greatly enhance the chemiluminescence emission of the luminol-H 2 O 2 system, and this finding resulted in a new chemiluminescence method for biosensing of glucose when coupled with the glucose oxidase.

  10. Cavitation occurrence around ultrasonic dental scalers.

    Science.gov (United States)

    Felver, Bernhard; King, David C; Lea, Simon C; Price, Gareth J; Damien Walmsley, A

    2009-06-01

    Ultrasonic scalers are used in dentistry to remove calculus and other contaminants from teeth. One mechanism which may assist in the cleaning is cavitation generated in cooling water around the scaler. The vibratory motion of three designs of scaler tip in a water bath has been characterised by laser vibrometry, and compared with the spatial distribution of cavitation around the scaler tips observed using sonochemiluminescence from a luminol solution. The type of cavitation was confirmed by acoustic emission analysed by a 'Cavimeter' supplied by NPL. A node/antinode vibration pattern was observed, with the maximum displacement of each type of tip occurring at the free end. High levels of cavitation activity occurred in areas surrounding the vibration antinodes, although minimal levels were observed at the free end of the tip. There was also good correlation between vibration amplitude and sonochemiluminescence at other points along the scaler tip. 'Cavimeter' analysis correlated well with luminol observations, suggesting the presence of primarily transient cavitation.

  11. Detection of gamma irradiated pepper and papain by chemiluminescence

    International Nuclear Information System (INIS)

    Sattar, Abdus; Delincee, H.; Diehl, J.F.

    1987-01-01

    Chemiluminescence (CL) measurements of black pepper and of papain using luminol and lucigenin reactions were studied. Effects of grinding, irradiation (5-20 kGy) and particle size (750-140 μm) on CL of pepper, and of irradiation (10-30 kGy) on CL of papain, were investigated. All the tested treatments affected the luminescence response in both the luminol and lucigenin reactions; however, the pattern of changes in each case, was inconsistent. Optimum pepper size for maximum luminescence was 560 μm, and optimum irradiation doses were > 15 kGy for pepper and > 20 kGy for papain. Chemiluminescence may possibly be used as an indicator or irradiation treatment for pepper and papain at a dose of 10 kGy or higher, but further research is needed to establish the reliability of this method. (author)

  12. Imaging Reactive Oxygen Species in Arthritis

    Directory of Open Access Journals (Sweden)

    Wei-Tsung Chen

    2004-07-01

    Full Text Available Reactive oxygen species (ROS have been shown to play a role in the pathogenesis of arthritides. Luminol was used as the primary reporter of ROS and photons resulting from the chemiluminescence reaction were detected using a super-cooled CCD photon counting system. Luminol was injected intravenously into groups of animals with different models of arthritis. Imaging signal correlated well with the severity of arthritis in focal and pan-arthritis as determined by histological measurement of ROS by formazan. Measurements were highly reproducible, sensitive, and repeatable. In vivo chemiluminescence imaging is expected to become a useful modality to elucidate the role of ROS in the pathogenesis of arthritides and in determining therapeutic efficacy of protective therapies.

  13. Applications of capillary electrophoresis with chemiluminescence detection in clinical, environmental and food analysis. A review

    International Nuclear Information System (INIS)

    Lara, Francisco J.; Airado-Rodríguez, Diego; Moreno-González, David; Huertas-Pérez, José F.; García-Campaña, Ana M.

    2016-01-01

    This paper reviews the latest developments and analytical applications of chemiluminescence detection coupled to capillary electrophoresis (CE-CL). Different sections considering the most common CL systems have been included, such as the tris(2,2′-bipyridine)ruthenium(II) system, the luminol and acridinium derivative reactions, the peroxyoxalate CL or direct oxidations. Improvements in instrumental designs, new strategies for improving both resolution and sensitivity, and applications in different fields such as clinical, pharmaceutical, environmental and food analysis have been included. This review covers the literature from 2010 to 2015. - Highlights: • An up-to-date critical review about the evolution of CE-CL is presented. • Tris(2,2′-bipyridine)ruthenium(II) and luminol as the most used CL systems. • Instrumental designs and strategies for improving resolution and sensitivity. • Applications in clinical, pharmaceutical, environmental and food analysis.

  14. Lyoluminescence of tris (hydroxymethyl) aminomethane in gamma dosimetry

    International Nuclear Information System (INIS)

    Azorin, J.; Gutierrez, A.; Guadarrama, L.

    1986-01-01

    Gamma dosimetry using lyoluminescence (LL) has advantages over other conventional systems because of its low cost and simplicity of use. Results of the investigation of the LL properties of tris (hydroxymethyl) aminomethane irradiated with 60 Co gamma radiation are presented and compared with the results of the LL of mannose and sucrose. Tris on dissolution in luminol shows a higher LL response than that of mannose and lower than that of sucrose, and has the highest stability of the three systems. The LL response of tris was linear in the range 5 Gy to 1kGy and showed no fading during one week. A comparison with the results of other authors as well as with measurements by ESR techniques are also reported. From these results we conclude that tris (hydroxymethyl) aminomethane on dissolution in luminol provides a LL dosimetric system useful for gamma dosimetry in the radiation processing of foods. (author)

  15. Antioxidant potential of six pine species.

    Science.gov (United States)

    Guri, Anilda; Kefalas, Panagiotis; Roussis, Vassilios

    2006-04-01

    The aim of the study was to evaluate the antioxidant efficacy of extracts obtained from six Pinus species (P. pinea, P. brutia, P. radiata, P. halepensis, P. attenuata, P. nigra) growing in natural forests in Southern Greece. Specimens of fresh, dry needles and pine bark were extracted and fractionated with a variety of organic solvents and the efficient concentration and their radical scavenging activity was evaluated by the Co(II)/EDTA induced luminol plateau chemiluminescence assay. Copyright 2006 John Wiley & Sons, Ltd.

  16. Sonochemical analysis of the output of ultrasonic dental descalers

    OpenAIRE

    King, David

    2010-01-01

    Ultrasonic descalers are used in dentistry to remove calculus and other contaminants from teeth. One mechanism which may assist in the cleaning is cavitation generated in cooling water around the descaler. The spatial distribution of cavitation around three designs of descaler tips and under three load conditions has been observed using sonochemiluminescence from a luminol solution and compared with the vibratory motion of the tips in a water bath, characterised by scanning laser vibrometry. ...

  17. Cavitation occurrence around ultrasonic dental scalers

    OpenAIRE

    Felver, Bernhard; King, David C; Lea, Simon C; Price, Gareth J; Damien Walmsley, A

    2009-01-01

    Ultrasonic scalers are used in dentistry to remove calculus and other contaminants from teeth. One mechanism which may assist in the cleaning is cavitation generated in cooling water around the scaler. The vibratory motion of three designs of scaler tip in a water bath has been characterised by laser vibrometry, and compared with the spatial distribution of cavitation around the scaler tips observed using sonochemiluminescence from a luminol solution. The type of cavitation was confirmed by a...

  18. Lyoluminescence technique as an identification method for irradiated food stuffs

    International Nuclear Information System (INIS)

    Chazhoor, J.S.

    1988-01-01

    The paper presents the studies made on the suitability of lyoluminescence technique as an analytical method for the identification of irradiated food stuffs. Powder milk, cinnamon, cardamom, clove, red chilly, cocoa, pepper, tea, coffee, turmeric and coriander showed lyoluminescence response when irradiated by a 10 kGy 60 Co and dissolved in luminol solution. Various dosimetric parameters such as effect of storage time, proportionality of the lyoluminescence response to dose etc were studied. (author). 1 tab., 3 figs

  19. Disposable, Autonomic, Energy-Converting Ion Channel Sensor Materials

    Science.gov (United States)

    2018-07-02

    a chemical reaction involving an analyte of interest results in light emission, is applicable to enzyme detection, DNA sequencing, treatment... chemical reaction itself. Most CL protocols use luminol, a well-known chemical that emits light when oxidized. The target analyte is typically labeled...Aziz Fennouri, Ph.D., Nanotechnology, Postdoctoral fellow Thomas Schroeder, Chemical Engineering , Ph.D. student Anirvan Guha, Physics, Ph.D. student

  20. Elimination of hydrogen peroxide by Haemophilus somnus, a catalase-negative pathogen of cattle.

    OpenAIRE

    Sample, A K; Czuprynski, C J

    1991-01-01

    Haemophilus somnus is a catalase-negative, gram-negative pathogen of cattle which is refractory to killing by bovine neutrophils. In this report, we showed that H. somnus rapidly inhibited Luminol-dependent chemiluminescence of bovine neutrophils costimulated with opsonized zymosan or phorbol myristate acetate. We have postulated that this inhibition resulted in part from H. somnus preventing the accumulation of hydrogen peroxide (H2O2) during the oxidative burst. In support of this hypothesi...

  1. Antioxidant activity of Citrus paradisi seeds glyceric extract.

    Science.gov (United States)

    Giamperi, Laura; Fraternale, Daniele; Bucchini, Anahi; Ricci, Donata

    2004-03-01

    The antioxidant activity of Citrus paradisi (grapefruit) seeds glyceric extract dissolved in ethanol and in aqueous media was evaluated using three different methods: evaluation by DPPH assay, by 5-lipoxygenase assay and by luminol/xanthine/xanthine oxidase chemiluminescence assay. The total phenolic content was determined by the Prussian Blue method opportunely modified. The grapefruit seeds glyceric extract utilized as aqueous solutions demonstrated antioxidant properties better than those displayed by alcoholic solutions.

  2. Potential role of NADPH-oxidase in early steps of lead-induced oxidative burst in Vicia faba roots

    OpenAIRE

    Pourrut, Bertrand; Perchet, Geoffrey; Silvestre, Jérôme; Cecchi, Marie; Guiresse, Agnès Maritchù; Pinelli, Eric

    2008-01-01

    The mechanism of oxidative burst induced by lead in Vicia faba excised roots was investigated by luminol-dependent chemiluminescence. Results showed that lead triggered a rapid and dose-dependent increase in chemiluminescence production. In this study, specific inhibitors of putative reactive oxygen species (ROS) sources were used to determine the mechanism of lead-induced ROS generation. This generation was sensitive to dephenylene iodonium (DPI), quinacrine and imidazole, some inhibitors of ...

  3. Gamma dosimetry using lyoluminescence of Tris (hydroxymethyl) aminomethane

    International Nuclear Information System (INIS)

    Mehta, S.K.; Oommen, I.K.; Sengupta, S.; Soman, S.D.

    1983-01-01

    Lyoluminescence (LL) of Tris (hydroxymethyl) aminomethane has been studied and compared with the LL of glucose, mannose, valine and glutamine. Among these materials, Tris (hydroxymethyl) aminomethane on dissolution in luminol shows the highest LL sensitivity and stability. The linear dosimetric range of this system is 0.1-2x10 2 Gy which makes it useful in the fields of radiotherapy and radiation processing of food. (orig.)

  4. Observation of chemiluminescence induced by hydrodynamic cavitation in microchannels.

    Science.gov (United States)

    Podbevsek, D; Colombet, D; Ledoux, G; Ayela, F

    2018-05-01

    We have performed hydrodynamic cavitation experiments with an aqueous luminol solution as the working fluid. Light emission, together with the high frequency noise which characterizes cavitation, was emitted by the two-phase flow, whereas no light emission from luminol was recorded in the single phase liquid flow. Light emission occurs downstream transparent microdiaphragms. The maximum level of the recorded signal was around 180 photons per second with flow rates of 380 µl/s, that corresponds to a real order of magnitude of the chemiluminescence of 75,000 photons per second. The yield of emitted photons increases linearly with the pressure drop, which is proportional to the square of the total flow rate. Chemiluminescence of luminol is a direct and a quantitative demonstration of the presence of OH hydroxyl radicals created by hydrodynamic cavitation. The presented method could be a key to optimize channel geometry for processes where radical production is essential. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. Enhanced visible-light-induced photocatalytic activity of α-Fe2O3 adsorbing redox enzymes

    Directory of Open Access Journals (Sweden)

    Kai Kamada

    2015-03-01

    Full Text Available We report fabrication of hybrid photocatalyst composed of an n-type semiconductor (α-Fe2O3 and a redox enzyme (horseradish peroxidase; HRP, and its performance for oxidation of luminol in an aqueous solution. The hybrid photocatalyst is simply formed via physical adsorption of HRP to an α-Fe2O3 sintered body. Under visible light irradiation, the bare α-Fe2O3 with a narrow bandgap photocatalytically oxidizes luminol along with blue emission that can be used as an indicator of the photocatalytic performance. The blue emission is largely strengthened after the adsorption of HRP, demonstrating that the presence of enzyme improves apparent photocatalytic activity of α-Fe2O3. The favorable effect is derived from synergistic oxidation of luminol by the biocatalysts (HRP as well as by the photocatalyst (α-Fe2O3. In this paper, influence of excitation wavelength, adsorption amount of HRP, and reaction temperature on the overall photocatalytic activity are elucidated, and then a reaction mechanism of the proposed novel hybrid photocatalyst is discussed in detail.

  6. A sniffer-camera for imaging of ethanol vaporization from wine: the effect of wine glass shape.

    Science.gov (United States)

    Arakawa, Takahiro; Iitani, Kenta; Wang, Xin; Kajiro, Takumi; Toma, Koji; Yano, Kazuyoshi; Mitsubayashi, Kohji

    2015-04-21

    A two-dimensional imaging system (Sniffer-camera) for visualizing the concentration distribution of ethanol vapor emitting from wine in a wine glass has been developed. This system provides image information of ethanol vapor concentration using chemiluminescence (CL) from an enzyme-immobilized mesh. This system measures ethanol vapor concentration as CL intensities from luminol reactions induced by alcohol oxidase and a horseradish peroxidase (HRP)-luminol-hydrogen peroxide system. Conversion of ethanol distribution and concentration to two-dimensional CL was conducted using an enzyme-immobilized mesh containing an alcohol oxidase, horseradish peroxidase, and luminol solution. The temporal changes in CL were detected using an electron multiplier (EM)-CCD camera and analyzed. We selected three types of glasses-a wine glass, a cocktail glass, and a straight glass-to determine the differences in ethanol emission caused by the shape effects of the glass. The emission measurements of ethanol vapor from wine in each glass were successfully visualized, with pixel intensity reflecting ethanol concentration. Of note, a characteristic ring shape attributed to high alcohol concentration appeared near the rim of the wine glass containing 13 °C wine. Thus, the alcohol concentration in the center of the wine glass was comparatively lower. The Sniffer-camera was demonstrated to be sufficiently useful for non-destructive ethanol measurement for the assessment of food characteristics.

  7. Graphene-based chemiluminescence resonance energy transfer for homogeneous immunoassay.

    Science.gov (United States)

    Lee, Joon Seok; Joung, Hyou-Arm; Kim, Min-Gon; Park, Chan Beum

    2012-04-24

    We report on chemiluminescence resonance energy transfer (CRET) between graphene nanosheets and chemiluminescent donors. In contrast to fluorescence resonance energy transfer, CRET occurs via nonradiative dipole-dipole transfer of energy from a chemiluminescent donor to a suitable acceptor molecule without an external excitation source. We designed a graphene-based CRET platform for homogeneous immunoassay of C-reactive protein (CRP), a key marker for human inflammation and cardiovascular diseases, using a luminol/hydrogen peroxide chemiluminescence (CL) reaction catalyzed by horseradish peroxidase. According to our results, anti-CRP antibody conjugated to graphene nanosheets enabled the capture of CRP at the concentration above 1.6 ng mL(-1). In the CRET platform, graphene played a key role as an energy acceptor, which was more efficient than graphene oxide, while luminol served as a donor to graphene, triggering the CRET phenomenon between luminol and graphene. The graphene-based CRET platform was successfully applied to the detection of CRP in human serum samples in the range observed during acute inflammatory stress.

  8. Design of Multi Bubble Sonoluminescence Reactor for Low Frequency Pressure Radiation

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Yun Seok; Lee, Jae Young [Handong Global University, Pohang (Korea, Republic of)

    2012-05-15

    Sonoluminescence phenomenon has been intensively studied due to its extraordinary capability to produce very high temperature and pressure within micro bubbles exposed by the pressure radiation. In general, it has been widely used for the chemical treatment including dissociation of the toxics and synthesis of functional materials such as nano catalysis. As for the nuclear applications, some of researchers tried to realization of the bubble fusion and change of the decay constant. Unfortunately applications to the nuclear industry are very skeptically accepted in the academic society. In spite of all such skepticism, the studies on sonoluminescence still have the room to be explored. In the present study, we investigated the relation among the reactor size, power and frequency of the pressure radiation. Main motivation of the present study came from some mismatch in the degradation rate of TCE in the multibubble sonoluminescence reactors (MBSL reactor) between Lee et al (2011) and Oh and Lee (2010). Both studies utilized horn type ultrasound source with the frequency of 20 kHz. However, the shape and volume of the reactors were different form each other. In the present study, we simply measured the light emission from the luminol solution in the reactors to evaluate the effectiveness of the MBSL. As noted in the study of Lee et al, the hot spot of MBSL dissociate water molecules into OH radicals which dissociate luminol in the solution to emit radiation. Therefore, the intensity and distribution of the radiation of luminol dissociation in the reactor are key index of the population of hot spots. Results and discussions are made by comparing the light emission intensity with different operating powers

  9. Multifunctional reduced graphene oxide trigged chemiluminescence resonance energy transfer: Novel signal amplification strategy for photoelectrochemical immunoassay of squamous cell carcinoma antigen.

    Science.gov (United States)

    Zhang, Yan; Sun, Guoqiang; Yang, Hongmei; Yu, Jinghua; Yan, Mei; Song, Xianrang

    2016-05-15

    Herein, a photoelectrochemical (PEC) immunoassay is constructed for squamous cell carcinoma antigen (SCCA) detection using zinc oxide nanoflower-bismuth sulfide (Bi2S3) composites as photoactive materials and reduced graphene oxide (rGO) as signal labels. Horseradish peroxidase is used to block sites against nonspecific binding, and then participated in luminol-based chemiluminescence (CL) system. The induced CL emission is acted as an inner light source to excite photoactive materials, simplifying the instrument. A novel signal amplification strategy is stem from rGO because of the rGO acts as an energy acceptor, while luminol serves as a donor to rGO, triggering the CL resonance energy transfer phenomenon between luminol and rGO. Thus, the efficient CL emission to photoactive materials decreases. Furthermore, the signal amplification caused by rGO labeled signal antibodies is related to photogenerated electron-hole pairs: perfect matching of energy levels between rGO and Bi2S3 makes rGO a sink to capture photogenerated electrons from Bi2S3; the increased steric hindrance hinders the electron donor to the surface of Bi2S3 for reaction with the photogenerated holes. On the basis of the novel signal amplification strategy, the proposed immunosensor exhibits excellent analytical performance for PEC detection of SCCA, ranging from 0.8 pg mL(-1) to 80 ng mL(-1) with a low detection limit of 0.21 pg mL(-1). Meanwhile, the designed signal amplification strategy provides a general format for future development of PEC assays. Copyright © 2015 Elsevier B.V. All rights reserved.

  10. Sequential Injection Determination of D-Glucose by Chemiluminescence Using an Open Tubular Immobilised Enzyme Reactor

    DEFF Research Database (Denmark)

    Liu, Xuezhu; Hansen, Elo Harald

    1996-01-01

    A sequential injection analysis system is described that incorporates a nylon tubular reactor containing immobilised glucose oxidase, allowing determination of D-glucose by means of subsequent luminol chemiluminescence detection of the hydrogen peroxide generated in the enzymatic reaction....... The operating parameters were optimised by fractional factorial screening and response surface modelling. The linear range of D-glucose determination was 30-600 mu M, With a detection limit of 15 mu M using a photodiode detector. The sampling frequency was 54 h(-1). Lower LOD (0.5 mu M D-glucose) could...

  11. On-line monitoring of Glucose and penicillin by sequential injection analysis

    DEFF Research Database (Denmark)

    Min, R.W.; Nielsen, Jens Bredal; Villadsen, John

    1996-01-01

    and a detector. The glucose analyzer is based on an enzymatic reaction using glucose oxidase, which converts glucose to glucono-lactone with formation of hydrogen peroxide and subsequent detection of H2O2 by a chemiluminescence reaction involving luminol. The penicillin analysis is based on formation......A sequential injection analysis (SIA) system has been developed for on-line monitoring of glucose and penicillin during cultivations of the filamentous fungus Penicillium chrysogenum. The SIA system consists of a peristaltic pump, an injection valve, two piston pumps, two multi-position valves...

  12. The synthesis of a water-soluble derivative of rutin as an antiradical agent

    Energy Technology Data Exchange (ETDEWEB)

    Pedriali, Carla Aparecida; Fernandes, Adjaci Uchoa [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Quimica. Dept. de Bioquimica]. E-mail: capedriali@hotmail.com; Bernusso, Leandra de Cassia; Polakiewicz, Bronislaw [Universidade de Sao Paulo (USP), SP (Brazil). Faculdade de Ciencias Farmaceuticas. Dept. de Tecnologia Bioquimico-Farmaceutica

    2008-07-01

    The purpose of this study was to synthesize a water-soluble derivative of rutin (compound 2) by introducing carboxylate groups on rutin's sugar moiety. The rutin derivative showed an almost 100-fold solubility increase in water. The antiradical capacity of compound 2 was evaluated using the luminol/AAPH system, and the derivative's activity was 1.5 times greater than that of Trolox. Despite the derivative's high solubility in water (log P = -1.13), lipid peroxidation of brain homogenate membranes was very efficiently inhibited (inhibition values were only 19% lower than the inhibition values of rutin). (author)

  13. Chemoluminescence measurement of 16 spices as a method to identify ionizing radiation treatment. Die Messung der Chemilumineszenz von 16 Gewuerzen als Nachweis einer Behandlung mit ionisierenden Strahlen

    Energy Technology Data Exchange (ETDEWEB)

    Heide, L; Boegl, W

    1984-09-01

    This report is based on a series of experiments conducted with 16 spices to find out whether the chemiluminescence intensity measurements of the reaction to luminol solution is appropriate as an indicator of treatment with ionizing radiation. For this purpose the optimum conditions of reaction were first assessed for each spice in order to determine the dependence of the chemiluminescence intensity on the radiation dose and storage time after irradiation. Only in garlic the irradiation was not able to be identified any more after 3 days of storage time. In most of the spices it is possible to take proof of radiation exposure even after storage times of more than 2 months.

  14. Lyoluminescence characteristics of lactose

    International Nuclear Information System (INIS)

    Oommen, I.K.; Sengupta, S.; Iyengar, T.S.

    1994-01-01

    The lyoluminescence (LL) of lactose shows a linear γ-ray dose response in the range 1-10 4 Gy. LL standardization measurement of lactose with a luminol sensitizer shows a sharp increase in light output in the pH range 10-11. Effects of radical scavengers and oxygen confirm that the radiation-induced free radicals take part in the LL process and oxygen is essential for initiating the LL mechanism. LL emission spectral measurements indicate an energy transfer from diffusing free radicals to sensitizer molecules to produce light. (author)

  15. A multisyringe flow injection system with immobilized glucose oxidase based on homogeneous chemiluminescence detection

    International Nuclear Information System (INIS)

    Manera, Matias; Miro, Manuel; Estela, Jose Manuel; Cerda, Victor

    2004-01-01

    In this paper, enzyme containing reactors are for the first time implemented in the multisyringe flow injection analysis (MSFIA) technique interfaced with chemiluminescence detection for biochemical assays. The automated methodology is based on the on-line substrate conversion in an oxidase packed-bed reactor and the post-column chemiluminogenic catalysed-reaction of the generated oxidising species with an organic molecule (namely, 3-aminophthalhydrazide) in front of the photosensor module. Various catalysts in homogeneous phase are compared taking advantage of the benefits of the MSFIA concept. On one hand, mineral catalysts (namely, Co(II)) are assessed, on the other hand, minute and accurate volumes of soluble organic species (viz., horseradish peroxidase (HRP)) are readily handled without requiring further immobilization protocols. The potentials of the MSFIA-CL concept with immobilisation of the proper oxidase protein are demonstrated using glucose as a model of substrate. Despite the different pH and kinetic requirements for both the substrate conversion in the enzyme-reactor and the Co(II)/HRP-mediated luminol oxidation integrated in the flow system, the MSFIA approach warrants maximum yields owing to the independent optimisation of the physical and chemical parameters of the various reactions involved. Under the optimised configurations and experimental variables, dynamic working ranges from 2.5x10 -6 to 1.0x10 -3 mol l -1 glucose may be obtained for both detection schemes by proper photomultiplier gain selection. The detection and determination limits calculated at the 3σ and 10σ level were 8.6x10 -7 and 2.0x10 -6 mol l -1 glucose, respectively, for the Co(II)-luminol system, and 1.3x10 -6 and 2.3x10 -6 mol l -1 glucose, respectively, for the HRP-luminol procedure. The repeatability (n=10) at the 1.0x10 -5 mol l -1 level was slightly better for the Co(II)-catalysed reaction (2.5% versus 4.0%). The developed MSFIA-CL methodology was used for kinetic

  16. The synthesis of a water-soluble derivative of rutin as an antiradical agent

    International Nuclear Information System (INIS)

    Pedriali, Carla Aparecida; Fernandes, Adjaci Uchoa; Bernusso, Leandra de Cassia; Polakiewicz, Bronislaw

    2008-01-01

    The purpose of this study was to synthesize a water-soluble derivative of rutin (compound 2) by introducing carboxylate groups on rutin's sugar moiety. The rutin derivative showed an almost 100-fold solubility increase in water. The antiradical capacity of compound 2 was evaluated using the luminol/AAPH system, and the derivative's activity was 1.5 times greater than that of Trolox. Despite the derivative's high solubility in water (log P = -1.13), lipid peroxidation of brain homogenate membranes was very efficiently inhibited (inhibition values were only 19% lower than the inhibition values of rutin). (author)

  17. Active oxygen doctors the evidence

    Science.gov (United States)

    Castelló, Ana; Francès, Francesc; Corella, Dolores; Verdú, Fernando

    2009-02-01

    Investigation at the scene of a crime begins with the search for clues. In the case of bloodstains, the most frequently used reagents are luminol and reduced phenolphthalein (or phenolphthalin that is also known as the Kastle-Meyer colour test). The limitations of these reagents have been studied and are well known. Household cleaning products have evolved with the times, and new products with active oxygen are currently widely used, as they are considered to be highly efficient at removing all kinds of stains on a wide range of surfaces. In this study, we investigated the possible effects of these new cleaning products on latent bloodstains that may be left at a scene of a crime. To do so, various fabrics were stained with blood and then washed using cleaning agents containing active oxygen. The results of reduced phenolphthalein, luminol and human haemoglobin tests on the washed fabrics were negative. The conclusion is that these new products alter blood to such an extent that it can no longer be detected by currently accepted methods employed in criminal investigations. This inability to locate bloodstains means that highly important evidence (e.g. a DNA profile) may be lost. Consequently, it is important that investigators are aware of this problem so as to compensate for it.

  18. Cavitation and non-cavitation regime for large-scale ultrasonic standing wave particle separation systems--In situ gentle cavitation threshold determination and free radical related oxidation.

    Science.gov (United States)

    Johansson, Linda; Singh, Tanoj; Leong, Thomas; Mawson, Raymond; McArthur, Sally; Manasseh, Richard; Juliano, Pablo

    2016-01-01

    We here suggest a novel and straightforward approach for liter-scale ultrasound particle manipulation standing wave systems to guide system design in terms of frequency and acoustic power for operating in either cavitation or non-cavitation regimes for ultrasound standing wave systems, using the sonochemiluminescent chemical luminol. We show that this method offers a simple way of in situ determination of the cavitation threshold for selected separation vessel geometry. Since the pressure field is system specific the cavitation threshold is system specific (for the threshold parameter range). In this study we discuss cavitation effects and also measure one implication of cavitation for the application of milk fat separation, the degree of milk fat lipid oxidation by headspace volatile measurements. For the evaluated vessel, 2 MHz as opposed to 1 MHz operation enabled operation in non-cavitation or low cavitation conditions as measured by the luminol intensity threshold method. In all cases the lipid oxidation derived volatiles were below the human sensory detection level. Ultrasound treatment did not significantly influence the oxidative changes in milk for either 1 MHz (dose of 46 kJ/L and 464 kJ/L) or 2 MHz (dose of 37 kJ/L and 373 kJ/L) operation. Copyright © 2015 Elsevier B.V. All rights reserved.

  19. Simultaneous determination of isoniazid and p-aminosalicylic acid by capillary electrophoresis using chemiluminescence detection.

    Science.gov (United States)

    Zhang, Xinfeng; Xuan, Yuelan; Sun, Aimin; Lv, Yi; Hou, Xiandeng

    2009-01-01

    It was found that isoniazid (ISO) or p-aminosalicylic acid (PAS) could enhance the chemiluminescence (CL) emission from Cu (II)-luminol-hydrogen peroxide system, and the increased chemiluminescence signals were proportional to their concentrations, respectively. Based on this phenomenon, a chemiluminescence method coupled to capillary electrophoresis (CE) was established for simultaneous determination of ISO and PAS. The CE conditions including running buffer and running voltage were investigated in detail. The effects of the pH of H(2)O(2) solution and the concentrations of luminol, H(2)O(2) and Cu (II) on the CL signal were also investigated carefully. Under the optimized conditions, the analysis could be accomplished within 10 min, with the limits of detection of 0.3 microg mL(-1) for ISO and 1.1 microg mL(-1) for PAS, corresponding to 7.2 and 26.4 pg per injection (24 nL), respectively. Finally, the method was validated by determining the two analytes in pharmaceutical preparation and spiked human serum samples. The results of pharmaceutical tablet analysis were in good agreement with the labeled amounts. The recoveries for ISO and PAS in human serum were in the range of 92-104% and 90-113%, respectively. Copyright 2008 John Wiley & Sons, Ltd.

  20. Effect of desensitizing agents on dentin permeability.

    Science.gov (United States)

    Ishihata, Hiroshi; Kanehira, Masafumi; Nagai, Tomoko; Finger, Werner J; Shimauchi, Hidetoshi; Komatsu, Masashi

    2009-06-01

    To investigate the in vitro efficacy of two dentin desensitizing products at reducing liquid permeability through human dentin discs. The tested hypothesis was that the products, in spite of different chemical mechanisms were not different at reducing or eliminating flow through dentin discs. Dentin slices (1 mm thick) were prepared from 16 extracted human third molars and their permeability was indirectly recorded in a split chamber model, using a chemiluminescence technique, after EDTA treatment (control), after soaking with albumin, and after desensitizer application. Two products were studied: MS Coat, a self-curing resin-containing oxalate product, and Gluma Desensitizer, a glutaraldehyde/HEMA-based agent without initiator. The dentin slices were mounted between an upper chamber, filled with an aqueous solution of 1% potassium ferricyanide and 0.3% hydrogen peroxide, and a lower chamber filled with 1% sodium hydroxide solution and 0.02% luminol. The upper solution was pressurized, and upon contact with the luminol solution a photochemical signal was generated and recorded as a measure of permeability throughout two consecutive pressurizing cycles at 2.5 and 13 kPa (26 and 133 cm H2O), respectively. The permeability of the control and albumin-soaked samples was similarly high. After application of the desensitizing agents, dentin permeability was reduced to virtually zero at both pressure levels (P < 0.001).

  1. On methods for the detection of reactive oxygen species generation by human spermatozoa: analysis of the cellular responses to catechol oestrogen, lipid aldehyde, menadione and arachidonic acid.

    Science.gov (United States)

    Aitken, R J; Smith, T B; Lord, T; Kuczera, L; Koppers, A J; Naumovski, N; Connaughton, H; Baker, M A; De Iuliis, G N

    2013-03-01

    Oxidative stress is known to have a major impact on human sperm function and, as a result, there is a need to develop sensitive methods for measuring reactive oxygen species (ROS) generation by these cells. A variety of techniques have been developed for this purpose including chemiluminescence (luminol and lucigenin), flow cytometry (MitoSOX Red, dihydroethidium, 4,5-diaminofluorescein diacetate and 2',7'-dichlorodihydrofluorescein diacetate) and spectrophotometry (nitroblue tetrazolium). The relative sensitivity of these assays and their comparative ability to detect ROS generated in different subcellular compartments of human spermatozoa, have not previously been investigated. To address this issue, we have compared the performance of these assays when ROS generation was triggered with a variety of reagents including 2-hydroxyestradiol, menadione, 4-hydroxynonenal and arachidonic acid. The results revealed that menadione predominantly induced release of ROS into the extracellular space where these metabolites could be readily detected by luminol-peroxidase and, to a lesser extent, 2',7'-dichlorodihydrofluorescein. However, such sensitivity to extracellular ROS meant that these assays were particularly vulnerable to interference by leucocytes. The remaining reagents predominantly elicited ROS generation by the sperm mitochondria and could be optimally detected by MitoSOX Red and DHE. Examination of spontaneous ROS generation by defective human spermatozoa revealed that MitoSOX Red was the most effective indicator of oxidative stress, thereby emphasizing the general importance of mitochondrial dysregulation in the aetiology of defective sperm function. © 2013 American Society of Andrology and European Academy of Andrology.

  2. Paramecium caudatum as a source of nitric oxide: Chemiluminescent detection based on Bluestar® Forensic reagent connected with microdialysis.

    Science.gov (United States)

    Bancirova, Martina

    2017-11-01

    Nitric oxide (NO) chemistry inside the body is the most interesting part of its behavior. NO is involved in controlling blood pressure, and in transmitting nerve signals and a variety of other signaling processes. To explain the behavior of NO, it is necessary to determine its immediate concentration or observe time-dependent changes in its concentration. In Paramecium caudatum, NO is formed by calcium-dependent nNOS (NOS1)-like protein, which is distributed in the cytoplasm. NO synthesis affects the ciliary beat and consequent motility of cells and blocked NO synthesis reduces the ability of cells to move. The possibility of online coupling of microdialysis (of P. caudatum solution) with NO detection is demonstrated. Direct measurement of NO is carried out using dilute Bluestar ® Forensic reagent (luminol-H 2 O 2 system; one of the NO detections is based upon the chemiluminescent reaction between NO and the luminol-H 2 O 2 system, which is specifically reactive to NO). The effect of a nitric oxide synthase inhibitor, NG-nitro-l-arginine methyl ester was observed. NO production was inhibited and the movement of P. caudatum was restricted. These effects were time dependent and after a specific time were reversed. Copyright © 2017 John Wiley & Sons, Ltd.

  3. Fructose and tagatose protect against oxidative cell injury by iron chelation.

    Science.gov (United States)

    Valeri, F; Boess, F; Wolf, A; Göldlin, C; Boelsterli, U A

    1997-01-01

    To further investigate the mechanism by which fructose affords protection against oxidative cell injury, cultured rat hepatocytes were exposed to cocaine (300 microM) or nitrofurantoin (400 microM). Both drugs elicited massively increased lactate dehydrogenase release. The addition of the ketohexoses D-fructose (metabolized via glycolysis) or D-tagatose (poor glycolytic substrate) significantly attenuated cocaine- and nitrofurantoin-induced cell injury, although both fructose and tagatose caused a rapid depletion of ATP and compromised the cellular energy charge. Furthermore, fructose, tagatose, and sorbose all inhibited in a concentration-dependent manner (0-16 mM) luminolenhanced chemiluminescence (CL) in cell homogenates, indicating that these compounds inhibit the iron-dependent reactive oxygen species (ROS)-mediated peroxidation of luminol. Indeed, both Fe2+ and Fe3+ further increased cocaine-stimulated CL, which was markedly quenched following addition of the ketohexoses. The iron-independent formation of superoxide anion radicals (acetylated cytochrome c reduction) induced by the prooxidant drugs remained unaffected by fructose or tagatose. The iron-chelator deferoxamine similarly protected against prooxidant-induced cell injury. In contrast, the nonchelating aldohexoses D-glucose and D-galactose did not inhibit luminol CL nor did they protect against oxidative cell injury. These data indicate that ketohexoses can effectively protect against prooxidant-induced cell injury, independent of their glycolytic metabolism, by suppressing the iron-catalyzed formation of ROS.

  4. DNA tetrahedral scaffolds-based platform for the construction of electrochemiluminescence biosensor.

    Science.gov (United States)

    Feng, Qiu-Mei; Zhou, Zhen; Li, Mei-Xing; Zhao, Wei; Xu, Jing-Juan; Chen, Hong-Yuan

    2017-04-15

    Proximal metallic nanoparticles (NPs) could quench the electrochemiluminescence (ECL) emission of semiconductor quantum dots (QDs) due to Förster energy transfer (FRET), but at a certain distance, the coupling of light-emission with surface plasmon resonance (SPR) result in enhanced ECL. Thus, the modification strategies and distances control between QDs and metallic NPs are critical for the ECL intensity of QDs. In this strategy, a SPR enhanced ECL sensor based on DNA tetrahedral scaffolds modified platform was reported for the detection of telomerase activity. Due to the rigid three-dimensional structure, DNA tetrahedral scaffolds grafting on the electrode surface could accurately modulate the distance between CdS QDs and luminol labelled gold nanoparticles (L-Au NPs), meanwhile provide an enhanced spatial dimension and accessibility for the assembly of multiple L-Au NPs. The ECL intensities of both CdS QDs (-1.25V vs. SCE) and luminol (+0.33V vs. SCE) gradually increased along with the formation of multiple L-Au NPs at the vertex of DNA tetrahedral scaffolds induced by telomerase, bringing in a dual-potential ECL analysis. The proposed method showed high sensitivity for the identification of telomerase and was successfully applied for the differentiation of cancer cells from normal cells. This work suggests that DNA tetrahedral scaffolds could serve as an excellent choice for the construction of SPR-ECL system. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Effect of resonance frequency, power input, and saturation gas type on the oxidation efficiency of an ultrasound horn.

    Science.gov (United States)

    Rooze, Joost; Rebrov, Evgeny V; Schouten, Jaap C; Keurentjes, Jos T F

    2011-01-01

    The sonochemical oxidation efficiency (η(ox)) of a commercial titanium alloy ultrasound horn has been measured using potassium iodide as a dosimeter at its main resonance frequency (20 kHz) and two higher resonance frequencies (41 and 62 kHz). Narrow power and frequency ranges have been chosen to minimise secondary effects such as changing bubble stability, and time available for radical diffusion from the bubble to the liquid. The oxidation efficiency, η(ox), is proportional to the frequency and to the power transmitted to the liquid (275 mL) in the applied power range (1-6 W) under argon. Luminol radical visualisation measurements show that the radical generation rate increases and a redistribution of radical producing zones is achieved at increasing frequency. Argon, helium, air, nitrogen, oxygen, and carbon dioxide have been used as saturation gases in potassium iodide oxidation experiments. The highest η(ox) has been observed at 5 W under air at 62 kHz. The presence of carbon dioxide in air gives enhanced nucleation at 41 and 62 kHz and has a strong influence on η(ox). This is supported by the luminol images, the measured dependence of η(ox) on input power, and bubble images recorded under carbon dioxide. The results give insight into the interplay between saturation gas and frequency, nucleation, and their effect on η(ox). Copyright © 2010 Elsevier B.V. All rights reserved.

  6. A chemiluminescent method for determination of lipid peroxidation

    International Nuclear Information System (INIS)

    Liang Xiaofeng; Hu Tianxi; Fan Xiaobing

    2003-01-01

    We established a chemiluminescent system for determination of lipid peroxidation and screening anti-oxidants. The lipid containing unsaturated fatly acids was injected into a galls tube. Luminol solution and the deionized water were added into it too. The glass tube was put into a preincubation box to incubate it for 0.5 h at 37 degree C. AAPH solution was injected into the tube for immediate measurement in a biochemiluminometer at 38-39 degree C. The pulses /6s(CP6s) were determined with T-2 program. Chemiluminescent dynamic and lipid peroxidation changes were observed continuously. Once the CL intensity of lipid peroxidation got peak, the antioxidant which has different concentration was added immediately in situ. A certain CL intensity (CP6s) was chosen as evaluation index to compare the activity of antioxidants. A luminol chemiluminescent system for determination of lipid peroxidation has been made. It was found that Vit. C, teapolyphenol, and glutathione have effects on scavenging lipid free radicals. The new method is quick, sensitive, and simple for determination of lipid peroxidation and screening antioxidants

  7. Quantum dots assisted photocatalysis for the chemiluminometric determination of chemical oxygen demand using a single interface flow system

    Energy Technology Data Exchange (ETDEWEB)

    Silvestre, Cristina I.C.; Frigerio, Christian [Requimte, Department of Chemistry, Faculty of Pharmacy, Porto University, Rua Anibal Cunha 164, 4099-030, Porto (Portugal); Santos, Joao L.M., E-mail: joaolms@ff.up.pt [Requimte, Department of Chemistry, Faculty of Pharmacy, Porto University, Rua Anibal Cunha 164, 4099-030, Porto (Portugal); Lima, Jose L.F.C. [Requimte, Department of Chemistry, Faculty of Pharmacy, Porto University, Rua Anibal Cunha 164, 4099-030, Porto (Portugal)

    2011-08-12

    Highlights: {yields} A novel flow method for the determination of chemical oxygen demand is proposed. {yields} CdTe nanocrystals are irradiated with UV light to generate strong oxidizing species. {yields} Reactive species promote a fast catalytic degradation of organic matter. {yields} Luminol is used as a chemiluminescence probe for indirect COD assessment. {yields} A single interface flow system was implemented to automate the assays. - Abstract: A novel flow method for the determination of chemical oxygen demand (COD) is proposed in this work. It relies on the combination of a fully automated single interface flow system, an on-line UV photocatalytic unit and quantum dot (QD) nanotechnology. The developed approach takes advantage of CdTe nanocrystals capacity to generate strong oxidizing species upon irradiation with UV light, which fostered a fast catalytic degradation of the organic compounds. Luminol was used as a chemiluminescence (CL) probe for indirect COD assessment, since it is easily oxidized by the QD generated species yielding a strong CL emission that is quenched in the presence of the organic matter. The proposed methodology allowed the determination of COD concentrations between 1 and 35 mg L{sup -1}, with good precision (R.S.D. < 1.1%, n = 3) and a sampling frequency of about 33 h{sup -1}. The procedure was applied to the determination of COD in wastewater certified reference materials and the obtained results showed an excellent agreement with the certified values.

  8. Application of multi-step excitation schemes for detection of actinides and lanthanides in solutions by luminescence/chemiluminescence laser spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Izosimov, I. [Joint Institute for Nuclear Research, Joliot Curie 6, Dubna 141980 (Russian Federation)

    2016-07-01

    The use of laser radiation with tunable wavelength allows the selective excitation of actinide/lanthanide species with subsequent registration of luminescence/chemiluminescence for their detection. This work is devoted to applications of the time-resolved laser-induced luminescence spectroscopy and time-resolved laser-induced chemiluminescence spectroscopy for the detection of lanthanides and actinides. Results of the experiments on U, Eu, and Sm detection by TRLIF (Time Resolved Laser Induced Fluorescence) method in blood plasma and urine are presented. Data on luminol chemiluminescence in solutions containing Sm(III), U(IV), and Pu(IV) are analyzed. It is shown that appropriate selectivity of lanthanide/actinide detection can be reached when chemiluminescence is initiated by transitions within 4f- or 5f-electron shell of lanthanide/actinide ions corresponding to the visible spectral range. In this case chemiluminescence of chemiluminogen (luminol) arises when the ion of f element is excited by multi-quantum absorption of visible light. The multi-photon scheme of chemiluminescence excitation makes chemiluminescence not only a highly sensitive but also a highly selective tool for the detection of lanthanide/actinide species in solutions. (author)

  9. Tape Cassette Bacteria Detection System

    Science.gov (United States)

    1973-01-01

    The design, fabrication, and testing of an automatic bacteria detection system with a zero-g capability and based on the filter-capsule approach is described. This system is intended for monitoring the sterility of regenerated water in a spacecraft. The principle of detection is based on measuring the increase in chemiluminescence produced by the action of bacterial porphyrins (i.e., catalase, cytochromes, etc.) on a luminol-hydrogen peroxide mixture. Since viable as well as nonviable organisms initiate this luminescence, viable organisms are detected by comparing the signal of an incubated water sample with an unincubated control. Higher signals for the former indicate the presence of viable organisms. System features include disposable sealed sterile capsules, each containing a filter membrane, for processing discrete water samples and a tape transport for moving these capsules through a processing sequence which involves sample concentration, nutrient addition, incubation, a 4 Molar Urea wash and reaction with luminol-hydrogen peroxide in front of a photomultiplier tube. Liquids are introduced by means of a syringe needle which pierces a rubber septum contained in the wall of the capsule. Detection thresholds obtained with this unit towards E. coli and S. marcescens assuming a 400 ml water sample are indicated.

  10. Ultrasound-mediated drug delivery by gas bubbles generated from a chemical reaction.

    Science.gov (United States)

    Lee, Sungmun; Al-Kaabi, Leena; Mawart, Aurélie; Khandoker, Ahsan; Alsafar, Habiba; Jelinek, Herbert F; Khalaf, Kinda; Park, Ji-Ho; Kim, Yeu-Chun

    2018-02-01

    Highly echogenic and ultrasound-responsive microbubbles such as nitrogen and perfluorocarbons have been exploited as ultrasound-mediated drug carriers. Here, we propose an innovative method for drug delivery using microbubbles generated from a chemical reaction. In a novel drug delivery system, luminol encapsulated in folate-conjugated bovine serum albumin nanoparticles (Fol-BSAN) can generate nitrogen gas (N 2 ) by chemical reaction when it reacts with hydrogen peroxide (H 2 O 2 ), one of reactive oxygen species (ROS). ROS plays an important role in the initiation and progression of cancer and elevated ROS have been observed in cancer cells both in vitro and in vivo. High-intensity focussed ultrasound (HIFU) is used to burst the N 2 microbubbles, causing site-specific delivery of anticancer drugs such as methotrexate. In this research, the drug delivery system was optimised by using water-soluble luminol and Mobil Composition of Matter-41 (MCM-41), a mesoporous material, so that the delivery system was sensitive to micromolar concentrations of H 2 O 2 . HIFU increased the drug release from Fol-BSAN by 52.9 ± 2.9% in 10 minutes. The cytotoxicity of methotrexate was enhanced when methotrexate is delivered to MDA-MB-231, a metastatic human breast cancer cell line, using Fol-BSAN with HIFU. We anticipate numerous applications of chemically generated microbubbles for ultrasound-mediated drug delivery.

  11. Involvement of both protein kinase C and G proteins in superoxide production after IgE triggering in guinea pig eosinophils

    Directory of Open Access Journals (Sweden)

    Toshiya Aizawa

    1997-01-01

    Full Text Available To study the function and mechanism of eosinophils via the low affinity IgE receptor (FceRII, we examined the production of 02 metabolites by measuring the luminol-dependent chemiluminescence (LDCL response and the generation of cysteinyl leukotrienes. Eosinophils obtained from guinea pig peritoneal fluid sensitized with horse serum were purified. Luminol-dependent chemiluminescence was induced by stimulation with monoclonal anti-CD23 antibody, but not by mouse serum (controls. The mean (±SEM value of LDCL was 20.6±1.3X103 c.p.m. This reaction consisted of an initial rapid phase and a propagation phase and ended within lOmin. Guinea pig eosinophils were histochemically stained with monoclonal anti-CD23 antibody. The major product generated in the LDCL response was superoxide, as determined by the measurement of superoxide by cytochrome c reduction and the complete inhibitory effect of superoxide dismutase on the LDCL response. Pretreatment with either pertussis toxin or cholera toxin inhibited the LDCL reaction. Depletion of bivalent ions by EDTA inhibited this response and the protein kinase C inhibitor D-sphingosin inhibited both 1-oleoyl-2-acetyl-glycerol-induced and FcϵRII-mediated LDCL. These findings suggest that the NADPH-protein kinase C pathway may be involved in the FceRII-mediated LDCL response in guinea pig eosinophils.

  12. Phytochemical Characteristics, Free Radical Scavenging Activities, and Neuroprotection of Five Medicinal Plant Extracts

    Directory of Open Access Journals (Sweden)

    Chia Lin Chang

    2012-01-01

    Full Text Available The objective of this study was to determine phytochemical characteristics, chemiluminescence antioxidant capacities, and neuroprotective effects on PC12 cells for methanol extracts of Spatholobus suberectus, Uncaria rhynchophylla, Alpinia officinarum, Drynaria fortunei, and Crataegus pinnatifida. The C. pinnatifida extract (CPE afforded the greatest yield and total phenolic content. The S. suberectus extract (SSE yielded the greatest total flavonoid content. The U. rhynchophylla extract (URE produced the greatest total tannin content, and the A. officinarum extract (AOE produced the greatest total triterpenoid content. The D. fortunei extract, assayed using horseradish peroxidase-luminol-hydrogen peroxide (H2O2, and AOE using pyrogallol-luminol assay each exhibited better antioxidant activity than the L-ascorbic acid and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid did. The CPE, SSE, and URE presented neurogrowth effects and neuroprotective activities on H2O2-induced PC12 cell death at 0.5–5.0 μg/mL. The CPE represents a promising medicinal plant source for the treatment of H2O2-induced neurodegenerative disease, because of its useful phytochemical characteristics.

  13. Phytochemical Characteristics, Free Radical Scavenging Activities, and Neuroprotection of Five Medicinal Plant Extracts

    Science.gov (United States)

    Chang, Chia Lin; Lin, Che San; Lai, Guia Hung

    2012-01-01

    The objective of this study was to determine phytochemical characteristics, chemiluminescence antioxidant capacities, and neuroprotective effects on PC12 cells for methanol extracts of Spatholobus suberectus, Uncaria rhynchophylla, Alpinia officinarum, Drynaria fortunei, and Crataegus pinnatifida. The C. pinnatifida extract (CPE) afforded the greatest yield and total phenolic content. The S. suberectus extract (SSE) yielded the greatest total flavonoid content. The U. rhynchophylla extract (URE) produced the greatest total tannin content, and the A. officinarum extract (AOE) produced the greatest total triterpenoid content. The D. fortunei extract, assayed using horseradish peroxidase-luminol-hydrogen peroxide (H2O2), and AOE using pyrogallol-luminol assay each exhibited better antioxidant activity than the L-ascorbic acid and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid did. The CPE, SSE, and URE presented neurogrowth effects and neuroprotective activities on H2O2-induced PC12 cell death at 0.5–5.0 μg/mL. The CPE represents a promising medicinal plant source for the treatment of H2O2-induced neurodegenerative disease, because of its useful phytochemical characteristics. PMID:21845204

  14. A molecularly imprinted electrochemiluminescence sensor based on the mimetic enzyme catalytic effect for ultra-trace Ni2+ determination.

    Science.gov (United States)

    Yang, Bin; Li, Jianping; Zhang, Lianming; Xu, Guobao

    2016-10-21

    A novel molecularly imprinted polymer (MIP) electrochemiluminescence (MIP-ECL) sensor was developed for the highly sensitive and selective determination of ultra-trace levels of Ni 2+ . The complex Ni 2+ -dimethylglyoxime (Ni-DMG) was chosen as the template molecule to construct the MIP and then acted as a mimetic enzyme to catalyse the oxidisation of luminol to enhance the ECL signal. When the imprinted cavities were occupied by Ni-DMG in the rebinding process, the ECL intensities produced by the luminol-H 2 O 2 ECL system on the MIP-modified electrode surface increased with increased concentration of the Ni-DMG complex. The highly sensitive determination of Ni 2+ was achieved through a catalytic reaction. This technique could be used for the quantitative analysis of Ni 2+ with concentrations from 3.0 × 10 -12 mol L -1 to 6.0 × 10 -9 mol L -1 . The detection limit was 1.01 × 10 -12 mol L -1 , which is much lower than that reported previously. In addition, the allowable amounts of interference ions in the MIP-ECL sensor were higher than that in other common molecularly imprinted sensors because of its excellent recognition of 3D cavity-to-complex molecules and ligand-to-metal ions. This method was successfully used to determine Ni 2+ in real samples, such as apples, carrots and grapes, and has been proven feasible for practical applications.

  15. Identification of irradiated spices with aid of scintillation counter

    International Nuclear Information System (INIS)

    Uusheimo, K.

    1989-08-01

    The aim off the work was to determine how one can identify gamma-irradiated spices with aid of a scintillation counter (LKB/Wallac 1219 RackBeta Spectral) by chemiluminescence measurements. Even though scintillation counters are more sensitive than real luminometers they have not been capable in identifying the irradiated spices after contact with photosensitizer like luminol, isoluminol and lucigenin presumably because the actual chemiluminescence reaction took place before the sample vial reached the measuring range. Whereas it was noticed that the identification of pure, dry allspice, black pepper, white peppar and cardemom was possible without any solutions when there were also present similar unirradiated spices. The identification was possible even after 23 weeks duration depending on the dose of the irradiation (10 kGy or 50 kGy) and the weight of the samples (1 g or 9 g). The duration of the investigation was 23 weeks

  16. Chemiluminescence immunoassay for chloramphenicol

    International Nuclear Information System (INIS)

    Lin Si; Xu Wenge; Liu Yibing

    2007-06-01

    A simple, solid-phase chemiluminescence immunoassay (CLIA) for the measurement of Chloramphenicol(CAP) in foodstuffs is described. A rabbit anti-CAP IgG is passively adsorbed onto the walls of polypropylene plates. The labeled conjugant is horseradish peroxidase(HRP) conjugate of CAP. Luminol solution is used as the substrate of HRP. The light yield is inversely proportional to the concentration of CAP. The method has a similar sensitivity (0.05 ng/mL), specificity, precision, and accuracy to a conventional enzyme immunoassay (EIA). The intra-assay and inter-assay CVs of ten samples were <8 and <20%, respectively, and the analytical recovery of the method was 87% 100%. The experimental correlation coefficient of dilution was found to be 0.999 using milk supernatant as buffer. The assay range for the method was 0.1-10 ng/mL, and it displayed good linearity. (authors)

  17. Potentiation of the generation of reactive oxidants by human phagocytes during exposure to benoxaprofen and ultraviolet radiation in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Anderson, R.; Eftychis, H.A.

    1986-09-01

    The effects of ultraviolet (UV) radiation on the spontaneous membrane-associated oxidative metabolism of human polymorphonuclear leukocytes (PMNL) and mononuclear leukocytes (MNL), co-incubated in the presence and absence of the non-steroidal, anti-inflammatory drug (NSAID) benoxaprofen at various concentrations, were investigated in vitro. Assays of superoxide generation and luminol-enhanced chemiluminescence (CL) were used to detect the production of reactive oxidants by PMNL and MNL. The pro-oxidative effects of benoxaprofen and UV radiation alone and in combination are dependent on intact phagocyte membrane-associated oxidative metabolism. It is postulated that the pro-oxidative interactions which occur between human phagocytes, benoxaprofen and ultraviolet radiation cause the dermatological side-effects of benoxaprofen.

  18. Superoxide scavenging activity of pirfenidone-iron complex

    International Nuclear Information System (INIS)

    Mitani, Yoshihiro; Sato, Keizo; Muramoto, Yosuke; Karakawa, Tomohiro; Kitamado, Masataka; Iwanaga, Tatsuya; Nabeshima, Tetsuji; Maruyama, Kumiko; Nakagawa, Kazuko; Ishida, Kazuhiko; Sasamoto, Kazumi

    2008-01-01

    Pirfenidone (PFD) is focused on a new anti-fibrotic drug, which can minimize lung fibrosis etc. We evaluated the superoxide (O 2 ·- ) scavenging activities of PFD and the PFD-iron complex by electron spin resonance (ESR) spectroscopy, luminol-dependent chemiluminescence assay, and cytochrome c reduction assay. Firstly, we confirmed that the PFD-iron complex was formed by mixing iron chloride with threefold molar PFD, and the complex was stable in distillated water and ethanol. Secondary, the PFD-iron complex reduced the amount of O 2 ·- produced by xanthine oxidase/hypoxanthine without inhibiting the enzyme activity. Thirdly, it also reduced the amount of O 2 ·- released from phorbor ester-stimulated human neutrophils. PFD alone showed few such effects. These results suggest the possibility that the O 2 ·- scavenging effect of the PFD-iron complex contributes to the anti-fibrotic action of PFD used for treating idiopathic pulmonary fibrosis

  19. Utilization of the human cell line HL-60 for chemiluminescence based detection of microorganisms and related substances

    DEFF Research Database (Denmark)

    Timm, Michael; Hansen, Erik W; Moesby, Lise

    2006-01-01

    species (ROS) when challenged with pyrogenic substances. In a luminol enhanced chemilumimetric assay the responsiveness of differentiated HL-60 cells is tested towards Salmonella typhimurium, Bacillus subtilis, Saccharomyces cerevisiae, Candida albicans, lipopolysaccharide (LPS) and lipoteichoic acid (LTA......). The results show a poor sensitivity to S. typhimurium but displays good sensitivity towards B. subtilis, LTA and LPS. Furthermore, the sensitivity towards the yeasts C. albicans and S. cerevisiae is considerably better than obtained in other in vitro cell systems. Overall these results indicate that the HL-60...... cell assay possibly could be evolved to a supplementary assay for the known pyrogenic detection assays. Furthermore, the utilization of the assay for pyrogenic examination of recombinant drugs derived from yeast expression systems would be relevant to examine....

  20. Chemiluminescence response of whole blood and separated blood cells in cases of experimentally induced pancreatitis and MDTQ-DA-Trasylol-ascorbic acid therapy

    Energy Technology Data Exchange (ETDEWEB)

    Zimmermann, T.; Schuster, R.; Lauschke, G.; Trausch, M. (Medical Academy of Dresden (Germany). Department of Surgery); Albrecht, S. (Medical Academy of Dresden (Germany). Institute of Clinical Chemistry); Kopprasch, S.; Kuehne, H. (Medical Academy of Dresden (Germany). Institute of Pathological Biochemistry)

    1991-12-24

    The formation of reactive O{sub 2} species in the pancreas of pigs after induction of necrotizing or oedematous pancreatitis was studied by means of luminol- and lucigenin-sensitized chemiluminescence. The effect of 2,2-dimethyl-4-methanesulphonic acid-1,2-dihydroquinoline in combination with Trasylol and ascorbic acid was studied in vivo. This combined therapy leads to a reduction in the chemiluminescence response by 50-70% with prevention of pancreatogenic shock and multiple organ failure by improvement of the gluthathione status. A combination of radical traps, kallikrein inhibitors and natural antioxidative sub-stances is an efficient alternative therapy in cases of acute pancrea-titis. (author). 10 refs.; 5 figs.

  1. Possibility of determination of the level of antioxidants in human body using spectroscopic methods

    International Nuclear Information System (INIS)

    Timofeeva, E; Gorbunova, E

    2016-01-01

    In this work, the processes of antioxidant defence against aggressive free radicals in human body were investigated theoretically; and the existing methods of diagnosis of oxidative stress and disturbance of antioxidant activity were reviewed. Also, the kinetics of free radical reactions in the oxidation of luminol and interaction antioxidants (such as chlorophyll in the multicomponent system of plant's leaves and ubiquinone) with the UV radiation were investigated experimentally by spectroscopic method. The results showed that this method is effective for recording the luminescence of antioxidants, free radicals, chemiluminescent reactions and fluorescence. In addition these results reveal new opportunities for the study of the antioxidant activity and antioxidant balance in a multicomponent system by allocating features of the individual components in spectral composition. A creation of quality control method for drugs, that are required for oxidative stress diagnosis, is a promising direction in the development of given work. (paper)

  2. The alpha hemolysin of Escherichia Coli power the metabolism oxidative of neutrophils human beings in response to the peptide chemotactic FMLP: comparison with the ionophore of calcium A23187

    International Nuclear Information System (INIS)

    Garcia, J.

    2000-01-01

    The calcium ionophore ionomycin primes polymorphonuclear leukocytes (PMN) for increased superoxide production upon stimulation with the chemotactic peptide FMLP (Helman Finkel, T. et al J Biol Chem 1987; 262: 12589-12596) In this investigation we assessed the effect of PMN priming with either alpha hemolysin (AH) or the calcium ionophore A23187, both of which increase intracellular calcium, on the oxidative metabolism of PMN (as measured by chemiluminescence) in response to secondary stimulation with FMLP. Both A23187 and AH priming increased, the luminol-enhanced chemiluminescence in response to secondary stimulation with FMLP, indicating overstimulation of PMLP oxidative metabolism. Additional experiments using lucigenin as chemiluminescence enhancer showed that A23187, but not AH priming of PMN, increased superoxide release in a manner similar to that reported for ionomycin. These results are discussed in reference to infectious processes involving hemolytic E. coli (Author) [es

  3. Chemoluminescence measurement of 16 spices as a method to identify ionizing radiation treatment

    International Nuclear Information System (INIS)

    Heide, L.; Boegl, W.

    1984-09-01

    This report is based on a series of experiments conducted with 16 spices to find out whether the chemiluminescence intensity measurements of the reaction to luminol solution is appropriate as an indicator of treatment with ionizing radiation. For this purpose the optimum conditions of reaction were first assessed for each spice in order to determine the dependence of the chemiluminescence intensity on the radiation dose and storage time after irradiation. Only in garlic the irradiation was not able to be identified any more after 3 days of storage time. In most of the spices it is possible to take proof of radiation exposure even after storage times of more than 2 months. (orig./PW) [de

  4. Irradiation monitoring of spices by chemiluminescence method

    International Nuclear Information System (INIS)

    Bhatti, K.A.; Khan, H.M.; Sattar, A.; Ahmad, A.

    2001-01-01

    Some spices (cumin, coriander, black pepper and red pepper), commonly used due to their specific aroma and nutritional value were studied using chemiluminescence (CL) method to test whether the food has been irradiated or not. The spices were ground to different mesh sizes and subjected to gamma ray doses of 2.5, 5.0, 7.5 and 10 Kgy. The effect of radiation doses and particle size on the CL response of irradiated and un-irradiated samples was checked using luminol and lucigenin sensitizers. The storage experiment was also performed for all spices with both the sensitizers in order to pick the suitable sensitizer for CL study. During this study Co-60 gamma ray source and Bio-orbit 1250 luminometer were used after precise calibration

  5. β-Orcinol Metabolites from the Lichen Hypotrachyna revoluta

    Directory of Open Access Journals (Sweden)

    Panagiota Papadopoulou

    2007-05-01

    Full Text Available Four new β-orcinol metabolites, hypotrachynic acid (1, deoxystictic acid (2, cryptostictinolide (3 and 8 ́-methylconstictic acid (4 along with the metabolites 8 ́-methylstictic acid (5, 8 ́-methylmenegazziaic acid (6, stictic acid (7, 8 ́-ethylstictic acid (8 and atranorin (9, that have been previously described, were isolated for the first time from the tissue extracts of the lichen Hypotrachyna revoluta (Flörke Hale. The structures of the new metabolites were elucidated on the basis of extensive spectroscopic analyses. Radical scavenging activity (RSA of the metabolites isolated in adequate amounts, was evaluated using luminol chemiluminescence and comparison with Trolox®.

  6. A novel flow injection chemiluminescence method for automated and miniaturized determination of phenols in smoked food samples.

    Science.gov (United States)

    Vakh, Christina; Evdokimova, Ekaterina; Pochivalov, Aleksei; Moskvin, Leonid; Bulatov, Andrey

    2017-12-15

    An easily performed fully automated and miniaturized flow injection chemiluminescence (CL) method for determination of phenols in smoked food samples has been proposed. This method includes the ultrasound assisted solid-liquid extraction coupled with gas-diffusion separation of phenols from smoked food sample and analytes absorption into a NaOH solution in a specially designed gas-diffusion cell. The flow system was designed to focus on automation and miniaturization with minimal sample and reagent consumption by inexpensive instrumentation. The luminol - N-bromosuccinimide system in an alkaline medium was used for the CL determination of phenols. The limit of detection of the proposed procedure was 3·10 -8 ·molL -1 (0.01mgkg -1 ) in terms of phenol. The presented method demonstrated to be a good tool for easy, rapid and cost-effective point-of-need screening phenols in smoked food samples. Copyright © 2017 Elsevier Ltd. All rights reserved.

  7. Characterization of the response chemiluminescence of neutrophils human beings to the hemolysin Escherichia coli alpha

    International Nuclear Information System (INIS)

    Garcia, J.

    2000-01-01

    Escherichia coli alpha hemolysin (AH) evoked a luminol-amplified chemiluminescence (CL) response from human polymorphonuclear leukocytes (PMN). Analysis of kinetic parameters of the PMN CL response to AH established similarities with that of PMN to the calcium ionophore A23187. PMN CL responses to both AH and A23187 were equally decreased by preincubating PMN with A63612, a hidroxamic acid derivative and lipooxigenase inhibitor, showing that the CL response to both hemolysin and ionophore share a common mechanism, probably activation of leukotriene synthesis, due to calcium entry into the cells brought about by AH and A23187. In addition, the CL response of PMN to AH was lowered by the hydroxyl radical scavenger dimethyl sulfoxide, further suggesting arachidonate metabolism is involved in CL response. (Author) [es

  8. New synthesis of gold nanocorals using a diazonium compound, and their application to an electrochemiluminescent assay of hydrogen peroxide

    International Nuclear Information System (INIS)

    Xu, Min; Qi, Wenjing; Zhang, Ling; Lai, Jianping; Aziz-ur-Rehman; Majeed, Saadat; Xu, Guobao

    2014-01-01

    The reaction of hydrogen tetracholoroaurate, sodium borohydride and the diazonium compound prepared from 4-aminobenzoic acid results in the formation of gold nanocorals (Au-NCs) for the first time. Scanning electron microscopy images and transmission electron microscopy images show that the Au-NCs are composed of nanowires with a diameter of 5.3 nm. A glassy carbon electrode modified with Au-NCs is found to trigger intense electrochemiluminescence of the luminol/H 2 O 2 system at a potential of −0.13 V. The effect was exploited to determine H 2 O 2 in the 0.1 to 100 μM concentration range with a 30 nM detection limit. (author)

  9. Luminescent chemical waves in the Cu(II)-catalyzed oscillatory oxidation of SCN- ions with hydrogen peroxide.

    Science.gov (United States)

    Pekala, Katarzyna; Jurczakowski, Rafał; Lewera, Adam; Orlik, Marek

    2007-05-10

    The oscillatory oxidation of thiocyanate ions with hydrogen peroxide, catalyzed by Cu2+ ions in alkaline media, was so far observed as occurring simultaneously in the entire space of the batch or flow reactor. We performed this reaction for the first time in the thin-layer reactor and observed the spatiotemporal course of the above process, in the presence of luminol as the chemiluminescent indicator. A series of luminescent patterns periodically starting from the random reaction center and spreading throughout the entire solution layer was reported. For a batch-stirred system, the bursts of luminescence were found to correlate with the steep decreases of the oscillating Pt electrode potential. These novel results open possibilities for further experimental and theoretical investigations of those spatiotemporal patterns, including studies of the mechanism of this chemically complex process.

  10. Chemiluminescence immunoassay for prostate-specific antigen

    International Nuclear Information System (INIS)

    Zhang Xuefeng; Liu Yibing; Jia Juanjuan; Xu Wenge; Li Ziying; Chen Yongli; Han Shiquan

    2008-01-01

    The chemiluminescence immunoassay (CLIA) for serum total prostate-specific antigen (T-PSA) was developed. The reaction of luminol with hydrogen peroxide was introduced into this chemiluminescence system. The detection limit is established as 0.12 μg/L (n=10, mean of zero standard + 2SD) and the analytical recovery of PSA is 83.8%-118.7%. The intra-assay and inter-assay CVs vary from 4.4%-5.0% and 6.2%-11.7%, respectively. The experimental correlation coefficient of dilution is found to be 0.999. Compared with immunoradiometric assay (IRMA) kits, the correlative equation is y=1.07x+0.68, and correlation coefficient r=0.97. The standard range for the method is 1.5-80 μg/L, and it presents good linearity. (authors)

  11. Phagocytic and chemiluminescent responses of mouse peritoneal macrophages to living and killed Salmonella typhimurium and other bacteria

    International Nuclear Information System (INIS)

    Tomita, T.; Blumenstock, E.; Kanegasaki, S.

    1981-01-01

    In the presence of luminol, resident as well as thioglycolate-induced and immunized macrophages emitted chemiluminescence more efficiently when the cells were exposed to living Salmonella typhimurium than when they were exposed to the same bacterium killed by ultraviolet light or heat. This phenomenon was observed whether or not the bacterium was opsonized. The different response to living and killed bacteria was also found with Escherichia coli, Pseudomonas aeruginosa, Proteus morganii, and Enterobacter aerogenes, but not with Shigella sonnei, Klebsiella pneumoniae, and Propionibacterium acnes. The results suggest that macrophages respond better to living, motile bacteria than to nonmotile or killed bacteria. The experimental results obtained with motility mutants of S. typhimurium, E. coli, and P. aeruginosa confirm that macrophages exposed to the motile bacteria emit chemiluminescence more efficiently and ingest the motile bacteria at a much faster rate than the nonmotile bacteria

  12. A low concentration of ethanol reduces the chemiluminescence of human granulocytes and monocytes but not the tumor necrosis factor alpha production by monocytes after endotoxin stimulation

    DEFF Research Database (Denmark)

    Parlesak, Alexandr; Diedrich, J. P.; Schäfer, Christian

    1998-01-01

    necrosis factor alpha (TNF-alpha) from Mphi. Further, the efficiency of ethanol to inactivate chemically generated ROS was tested. Significant stimulation of ROS release occurred at endotoxin concentrations of 1 ng/ml or higher in both PMNs and Mphi. Ethanol significantly suppressed the formation of ROS...... immunogens and to increase the susceptibility of alcohol abusers to infectious diseases. As endotoxemia is common in alcohol abusers, we investigated the effect of ethanol (21.7 mmol/liter) on the luminol-amplified chemiluminescence of PMNs and Mphi after endotoxin stimulation and the release of tumor...... identical (6 to 8 ng/ml) in both PMNs and Mphi, independent of the presence of ethanol. In contrast to ROS formation, ethanol had no effect on the amount of TNF-alpha produced by endotoxin-stimulated Mphi. Ethanol was shown to be unable to decrease the levels of chemically generated ROS under physiological...

  13. The antioxidants curcumin and quercetin inhibit inflammatory processes associated with arthritis.

    Science.gov (United States)

    Jackson, J K; Higo, T; Hunter, W L; Burt, H M

    2006-04-01

    Curcumin and quercetin are antioxidant molecules with anti-proliferative, anti-inflammatory and immunosuppressive activities. The objective of this study was to investigate the inhibitory activity of these agents using four assays of inflammatory aspects of arthritis. Crystal-induced neutrophil activation was measured by luminol-dependent chemiluminescence. Synoviocyte proliferation was measured by an MTS assay using HIG-82 rabbit synoviocytes in cell culture. Chondrocyte (cultured primary cells) expression of the matrix metalloproteinases collagenase and stromelysin was measured by Northern Blot analysis. Angiogenesis was measured using the chorioallantoic membrane of the chick embryo. Both agents inhibited neutrophil activation, synoviocyte proliferation and angiogenesis. Curcumin strongly inhibited collagenase and stromelysin expression at micromolar concentrations whereas quercetin had no effect in this assay. These studies suggest that curcumin and to a lesser extent quercetin may offer therapeutic potential for the treatment of crystal-induced arthritis or rheumatoid arthritis.

  14. An electrochemiluminescence-based fibre optic biosensor for choline flow injection analysis.

    Science.gov (United States)

    Tsafack, V C; Marquette, C A; Leca, B; Blum, L J

    2000-01-01

    A fibre optic biosensor based on luminol electrochemiluminescence (ECL) integrated in a flow injection analysis (FIA) system was developed for the detection of choline. The electrochemiluminescence of luminol was generated by a glassy carbon electrode polarised at +425 mV vs. a platinum pseudo-reference electrode. Choline oxidase (Chx) was immobilised either covalently on polyamide (ABC type) or on UltraBind preactivated membranes, or by physical entrapment in a photo-cross-linkable poly(vinyl alcohol) polymer (PVA-SbQ) alone or after absorption on a weak anion exchanger, DEAE (diethylaminoethyl) Sepharose. The optimisation of the reaction conditions and physicochemical parameters influencing the FIA biosensor response demonstrated that the choline biosensor exhibited the best performances in a 30 mM veronal buffer containing 30 mM KCl and 1.5 mM MgCl2, at pH 9. The use of a 0.5 ml min-1 flow rate enabled the measurement of choline by the membrane-based ECL biosensors in 8 or 5 min, with ABC or UltraBind membranes, respectively, whereas the measurement required only 3 min with the DEAE-PVA system. For comparison, the detection of choline was performed with Chx immobilised using the four different supports. The best performances were obtained with the DEAE-PVA-Chx sensing layer, which allowed a detection limit of 10 pmol, whereas with the ABC, the UltraBind and the PVA systems, the detection limits were 300 pmol, 75 pmol and 220 pmol, respectively. The DEAE-based system also exhibited a good operational stability since 160 repeated measurements of 3 nmol of choline could be performed with an RSD of 4.5% whereas the stability under the best conditions was 45 assays with the other supports.

  15. Chitosan oligosaccharides attenuates oxidative-stress related retinal degeneration in rats.

    Directory of Open Access Journals (Sweden)

    I-Mo Fang

    Full Text Available This study investigated the therapeutic potential and mechanisms of chitosan oligosaccharides (COS for oxidative stress-induced retinal diseases. Retinal oxidative damage was induced in Sprague-Dawley rats by intravitreal injection of paraquat (PQ. Low-dose (5 mg/kg or high-dose (10 mg/kg COS or PBS was intragastrically given for 14 days after PQ injection. Electroretinograms were performed to determine the functionality of the retinas. The surviving neurons in the retinal ganglion cell layer and retinal apoptosis were determined by counting Neu N-positive cells in whole-mounted retinas and TUNEL staining, respectively. The generation of reactive oxygen species (ROS was determined by lucigenin- and luminol-enhanced chemiluminescence. Retinal oxidative damages were assessed by staining with nitrotyrosine, acrolein, and 8-hydroxy-2'-deoxyguanosine (8-OHdG. Immunohistochemical studies were used to demonstrate the expression of nuclear factor-kappa B (NF-κB p65 in retinas. An in vitro study using RGC-5 cells was performed to verify the results. We demonstrated COS significantly enhanced the recovery of retinal function, preserved inner retinal thickness, and decreased retinal neurons loss in a dose-dependent manner. COS administration demonstrated anti-oxidative effects by reducing luminol- and lucigenin-dependent chemiluminenscense levels and activating superoxide dismutase and catalase, leading to decreased retinal apoptosis. COS markedly reduced retinal NF-κB p65. An in vitro study demonstrated COS increased IκB expression, attenuated the increase of p65 and thus decreased NF-κB/DNA binding activity in PQ-stimulated RGC-5 cells. In conclusion, COS attenuates oxidative stress-induced retinal damages, probably by decreasing free radicals, maintaining the activities of anti-oxidative enzymes, and inhibiting the activation of NF-κB.

  16. An ultrasensitive chemiluminescence immunoassay for fumonisin B1 detection in cereals based on gold-coated magnetic nanoparticles.

    Science.gov (United States)

    Jie, Mingsha; Yu, Songcheng; Yu, Fei; Liu, Lie; He, Leiliang; Li, Yanqiang; Zhang, Hongquan; Qu, Lingbo; Harrington, Peter de B; Wu, Yongjun

    2018-07-01

    In the present study, a novel highly sensitive magnetic enzyme chemiluminescence immunoassay (MECLIA) was developed to detect fumonisin B 1 (FB 1 ) in cereal samples. The gold-coated magnetic nanoparticles (Fe 3 O 4 @Au, GoldMag) were used as solid phase carrier to develop a competitive CLIA for detecting FB 1 , in which FB 1 in samples would compete with FB 1 -ovalbumin coated on the surface of Fe 3 O 4 @Au nanoparticles for binding with FB 1 antibodies. Successively, horseradish peroxidase labeled goat anti-rabbit IgG (HRP-IgG) was conjugated with FB 1 antibodies on the microplate. In substrate solution containing luminol and H 2 O 2 , HRP-IgG catalyzed luminol oxidation by H 2 O 2 , generating a high chemiluminescence signal. The FB 1 immune GoldMag particles were characterized by Fourier transform infrared spectroscopy, scanning electron microscope and zeta potential analysis, etc. RESULTS: The concentrations and the reaction times of these immunoreagents were optimized to improve the performances of this method. The established method could detect as low as 0.027 ng mL -1 FB 1 from 0.05 ng mL -1 to 25 ng mL -1 , demonstrating little cross-reaction (less than 2.4%) with other structurally related compounds. The average intrassay relative SD (RSD) (n = 6) was 3.4% and the average interassay RSD (n = 6) was 5.4%. This method was successfully applied for the determination of FB 1 in corn and wheat and gave recoveries of between 98-110% and 91-105%, respectively. The results of the present study suggest that the MECLIA approach has potential application for high-throughput fumonisin screening in cereals. © 2018 Society of Chemical Industry. © 2018 Society of Chemical Industry.

  17. Effect of therapeutic plasma concentrations of non-steroidal anti-inflammatory drugs on the production of reactive oxygen species by activated rat neutrophils

    Directory of Open Access Journals (Sweden)

    Paino I.M.M.

    2005-01-01

    Full Text Available The release of reactive oxygen specie (ROS by activated neutrophil is involved in both the antimicrobial and deleterious effects in chronic inflammation. The objective of the present investigation was to determine the effect of therapeutic plasma concentrations of non-steroidal anti-inflammatory drugs (NSAIDs on the production of ROS by stimulated rat neutrophils. Diclofenac (3.6 µM, indomethacin (12 µM, naproxen (160 µM, piroxicam (13 µM, and tenoxicam (30 µM were incubated at 37ºC in PBS (10 mM, pH 7.4, for 30 min with rat neutrophils (1 x 10(6 cells/ml stimulated by phorbol-12-myristate-13-acetate (100 nM. The ROS production was measured by luminol and lucigenin-dependent chemiluminescence. Except for naproxen, NSAIDs reduced ROS production: 58 ± 2% diclofenac, 90 ± 2% indomethacin, 33 ± 3% piroxicam, and 45 ± 6% tenoxicam (N = 6. For the lucigenin assay, naproxen, piroxicam and tenoxicam were ineffective. For indomethacin the inhibition was 52 ± 5% and diclofenac showed amplification in the light emission of 181 ± 60% (N = 6. Using the myeloperoxidase (MPO/H2O2/luminol system, the effects of NSAIDs on MPO activity were also screened. We found that NSAIDs inhibited both the peroxidation and chlorinating activity of MPO as follows: diclofenac (36 ± 10, 45 ± 3%, indomethacin (97 ± 2, 100 ± 1%, naproxen (56 ± 8, 76 ± 3%, piroxicam (77 ± 5, 99 ± 1%, and tenoxicam (90 ± 2, 100 ± 1%, respectively (N = 3. These results show that therapeutic levels of NSAIDs are able to suppress the oxygen-dependent antimicrobial or oxidative functions of neutrophils by inhibiting the generation of hypochlorous acid.

  18. In Vivo Bioluminescence Imaging for Longitudinal Monitoring of Inflammation in Animal Models of Uveitis.

    Science.gov (United States)

    Gutowski, Michal B; Wilson, Leslie; Van Gelder, Russell N; Pepple, Kathryn L

    2017-03-01

    We develop a quantitative bioluminescence assay for in vivo longitudinal monitoring of inflammation in animal models of uveitis. Three models of experimental uveitis were induced in C57BL/6 albino mice: primed mycobacterial uveitis (PMU), endotoxin-induced uveitis (EIU), and experimental autoimmune uveitis (EAU). Intraperitoneal injection of luminol sodium salt, which emits light when oxidized, provided the bioluminescence substrate. Bioluminescence images were captured by a PerkinElmer In Vivo Imaging System (IVIS) Spectrum and total bioluminescence was analyzed using Living Image software. Bioluminescence on day zero was compared to bioluminescence on the day of peak inflammation for each model. Longitudinal bioluminescence imaging was performed in EIU and EAU. In the presence of luminol, intraocular inflammation generates detectable bioluminescence in three mouse models of uveitis. Peak bioluminescence in inflamed PMU eyes (1.46 × 105 photons/second [p/s]) was significantly increased over baseline (1.47 × 104 p/s, P = 0.01). Peak bioluminescence in inflamed EIU eyes (3.18 × 104 p/s) also was significantly increased over baseline (1.09 × 104 p/s, P = 0.04), and returned to near baseline levels by 48 hours. In EAU, there was a nonsignificant increase in bioluminescence at peak inflammation. In vivo bioluminescence may be used as a noninvasive, quantitative measure of intraocular inflammation in animal models of uveitis. Primed mycobacterial uveitis and EIU are both acute models with robust anterior inflammation and demonstrated significant changes in bioluminescence corresponding with peak inflammation. Experimental autoimmune uveitis is a more indolent posterior uveitis and generated a more modest bioluminescent signal. In vivo imaging system bioluminescence is a nonlethal, quantifiable assay that can be used for monitoring inflammation in animal models of uveitis.

  19. Photoactivation by visible light of CdTe quantum dots for inline generation of reactive oxygen species in an automated multipumping flow system

    Energy Technology Data Exchange (ETDEWEB)

    Ribeiro, David S.M.; Frigerio, Christian; Santos, Joao L.M. [Requimte, Department of Chemical Sciences, Laboratory of Applied Chemistry, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira no. 228, 4050-313 Porto (Portugal); Prior, Joao A.V., E-mail: joaoavp@ff.up.pt [Requimte, Department of Chemical Sciences, Laboratory of Applied Chemistry, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira no. 228, 4050-313 Porto (Portugal)

    2012-07-20

    Highlights: Black-Right-Pointing-Pointer CdTe quantum dots generate free radical species upon exposure to visible radiation. Black-Right-Pointing-Pointer A high power visible LED lamp was used as photoirradiation element. Black-Right-Pointing-Pointer The laboratory-made LED photocatalytic unit was implemented inline in a MPFS. Black-Right-Pointing-Pointer Free radical species oxidize luminol producing a strong chemiluminescence emission. Black-Right-Pointing-Pointer Epinephrine scavenges free radical species quenching chemiluminescence emission. - Abstract: Quantum dots (QD) are semiconductor nanocrystals able to generate free radical species upon exposure to an electromagnetic radiation, usually in the ultraviolet wavelength range. In this work, CdTe QD were used as highly reactive oxygen species (ROS) generators for the control of pharmaceutical formulations containing epinephrine. The developed approach was based on the chemiluminometric monitoring of the quenching effect of epinephrine on the oxidation of luminol by the produced ROS. Due to the relatively low energy band-gap of this chalcogenide a high power visible light emitting diode (LED) lamp was used as photoirradiation element and assembled in a laboratory-made photocatalytic unit. Owing to the very short lifetime of ROS and to ensure both reproducible generation and time-controlled reaction implementation and development, all reactional processes were implemented inline by using an automated multipumping micro-flow system. A linear working range for epinephrine concentration of up to 2.28 Multiplication-Sign 10{sup -6} mol L{sup -1} (r = 0.9953; n = 5) was verified. The determination rate was about 79 determinations per hour and the detection limit was about 8.69 Multiplication-Sign 10{sup -8} mol L{sup -1}. The results obtained in the analysis of epinephrine pharmaceutical formulations by using the proposed methodology were in good agreement with those furnished by the reference procedure, with

  20. Fingerprint enhancement revisited and the effects of blood enhancement chemicals on subsequent profiler Plus fluorescent short tandem repeat DNA analysis of fresh and aged bloody fingerprints.

    Science.gov (United States)

    Frégeau, C J; Germain, O; Fourney, R M

    2000-03-01

    This study was aimed at determining the effect of seven blood enhancement reagents on the subsequent Profiler Plus fluorescent STR DNA analysis of fresh or aged bloody fingerprints deposited on various porous and nonporous surfaces. Amido Black, Crowle's Double Stain. 1,8-diazafluoren-9-one (DFO), Hungarian Red, leucomalachite green, luminol and ninhydrin were tested on linoleum, glass, metal, wood (pine, painted white), clothing (85% polyester/15% cotton, 65% polyester/35% cotton, and blue denim) and paper (Scott 2-ply and Xerox-grade). Preliminary experiments were designed to determine the optimal blood dilutions to use to ensure a DNA typing result following chemical enhancement. A 1:200 blood dilution deposited on linoleum and enhanced with Crowle's Double Stain generated enough DNA for one to two rounds of Profiler Plus PCR amplification. A comparative study of the DNA yields before and after treatment indicated that the quantity of DNA recovered from bloody fingerprints following enhancement was reduced by a factor of 2 to 12. Such a reduction in the DNA yields could potentially compromise DNA typing analysis in the case of small stains. The blood enhancement chemicals selected were also evaluated for their capability to reveal bloodmarks on the various porous and nonporous surfaces chosen in this study. Luminol. Amido Black and Crowle's Double Stain showed the highest sensitivity of all seven chemicals tested and revealed highly diluted (1:200) bloody fingerprints. Both luminol and Amido Black produced excellent results on both porous and nonporous surfaces, but Crowle's Double Stain failed to produce any results on porous substrates. Hungarian Red, DFO, leucomalachite green and ninhydrin showed lower sensitivities. Enhancement of bloodmarks using any of the chemicals selected, and short-term exposure to these same chemicals (i.e., less than 54 days), had no adverse effects on the PCR amplification of the nine STR systems surveyed (D3S 1358, HumvWA, Hum

  1. Sequential injection chemiluminescence immunoassay for nonionic surfactants by using magnetic microbeads

    International Nuclear Information System (INIS)

    Zhang Ruiq; Nakajima, Hizuru; Soh, Nobuaki; Nakano, Koji; Masadome, Takashi; Nagata, Kazumi; Sakamoto, Kazuhira; Imato, Toshihiko

    2007-01-01

    A rapid and sensitive immunoassay based on a sequential injection analysis (SIA) using magnetic microbeads for the determination of alkylphenol polyethoxylates (APnEOs) is described. An SIA system was constructed from a syringe pump, a switching valve, a flow-through type immunoreaction cell equipped with a photon counting unit and a neodymium magnet. Magnetic beads, to which an anti-APnEOs monoclonal antibody was immobilized, were used as a solid support in an immunoassay. The introduction, trapping and release of the magnetic beads in and from the immunoreaction cell were controlled by means of a neodymium magnet and adjusting the flow of a carrier solution. The immunoassay was based on an indirect competitive immunoreaction of an anti-APnEOs monoclonal antibody immobilized on the magnetic beads with a sample APnEOs and a horseradish peroxidase (HRP)-labeled APnEOs in the same sample solution, and was based on the subsequent chemiluminscence reaction of HRP on the magnetic microbeads with a luminol solution containing hydrogen peroxide and p-iodophenol. The anti-APnEOs antibody was immobilized on the magnetic microbeads by coupling the antibody with the magnetic beads after activation of a carboxylate moiety on the surface of the magnetic beads that had been coated with a polylactic acid film. The antibody immobilized magnetic beads were introduced in the immunoreaction cell and trapped in it by the neodymium magnet, which was equipped beneath the immunoreaction cell. An APnEOs sample solution containing the HRP-labeled APnEOs at a constant concentration, and a luminol solution containing hydrogen peroxide and p-iodophenol were sequentially introduced into the immunoreaction cell, according to an SIA programmed sequence. Chemiluminescence emission was monitored by means of a photon counting unit located at the upper side of the immunoreaction cell by collecting the emitted light with a lens. A typical sigmoidal calibration curve was obtained, when the logarithm

  2. Flow-through solid-phase based optical sensor for the multisyringe flow injection trace determination of orthophosphate in waters with chemiluminescence detection

    International Nuclear Information System (INIS)

    Morais, Ines P.A.; Miro, Manuel; Manera, Matias; Estela, Jose Manuel; Cerda, Victor; Souto, M. Renata S.; Rangel, Antonio O.S.S.

    2004-01-01

    In this work, a novel flow-through solid-phase based chemiluminescence (CL) optical sensor is described for the trace determination of orthophosphate in waters exploiting the multisyringe flow injection analysis (MSFIA) concept with multicommutation. The proposed time-based injection flow system relies upon the in-line derivatisation of the analyte with ammonium molybdate in the presence of vanadate, and the transient immobilisation of the resulting heteropolyacid in a N-vinylpyrrolidone/divinylbenzene copolymer packed spiral shape flow-through cell located in front of the window of a photomultiplier tube. The simultaneous injection of well-defined slugs of luminol in alkaline medium and methanol solution towards the packed reactor is afterwards performed by proper switching of the solenoid valves. Then, the light emission from the luminol oxidation by the oxidant species retained onto the sorbent material is readily detected. At the same time, the generated molybdenum-blue compound is eluted by the minute amount of injected methanol, rendering the system prepared for a new measuring cycle. Therefore, the devised sensor enables the integration of the solid-phase CL reaction with elution and detection of the emitted light without the typical drawbacks of the molybdenum-blue based spectrophotometric procedures regarding the excess of molybdate anion, which causes high background signals due to its self-reduction. The noteworthy features of the developed CL-MSFIA system are the feasibility to accommodate reactions with different pH requirements and the ability to determine trace levels of orthophosphate in high silicate content samples (Si/P ratios up to 500). Under the optimised conditions, a dynamic linear range from 5 to 50 μg P l -1 for a 1.8 ml sample, repeatability better than 3.0% and a quantification limit of 4 μg P l -1 were attained. The flowing stream system handles 11 analysis h -1 and has been successfully applied to the determination of trace levels of

  3. Estimating the yin-yang nature of Western herbs: a potential tool based on antioxidation-oxidation theory.

    Science.gov (United States)

    Gilca, Marilena; Gaman, Laura; Lixandru, Daniela; Stoian, Irina

    2014-01-01

    One of the biggest obstacles to progress in traditional Chinese medicine (TCM) development in Western countries is the difficulty of applying the traditional concepts to the Western medicinal plants, which are not traditionally described in ancient literature. During recent years, new advances in the field of understanding Yin/Yang aspects from a modern bioscientific point of view have led to the conclusion that antioxidationoxidation concepts might mirror a Yin-Yang relationship. This study was intended to integrate the Yin-Yang theory of the traditional Chinese medicine with modern antioxidation-oxidation theory, and to propose a biochemical tool based on redox parameters (e.g. antioxidant capacity, chemiluminescence-CL signal inducing capacity), usable for the classification of Western medicinal plants from Yin/Yang perspective. Trolox equivalent antioxidant capacity (TEAC) of six vegetal aqueous extracts (Symphitum officinalae (radix)-SYM, Inula helenium (radix)-INU, Calendula officinalis (flores)-CAL, Angelica arhanghelica (folium)ANG(F), Angelica arhanghelica (radix)-ANG(R), Ecbalium Elaterium (fruits)-ECB) and luminol-enhanced chemiluminescence of PMNL on addition of these vegetal extracts were measured. Percentages from the maximal or minimal values obtained were calculated for each extract (TEAC%, PMNL stimulation%, PMNL inhibition%, relative speed of action% (RSA%%)), specific Yin-Yang significance was assigned to each relative parameter. In the end, an integration of all the relative values was done, in order to find a global "Yin" or a "Yang" trait of each vegetal extract. TEAC decreased in the following order: SYM > INU > CAL >ANG(F) > ANG(R > ECB. Three vegetal extracts (SYM > INU > ECB) decreased the luminol-enhanced chemiluminescence of PMNL, two (ANG(R) > ANG(F)) increased it, while one (CAL) had a dual effect. After the integration of the percentages, CAL was found to have a global "Yang" trait, while the rest of the plants had a global "Yin

  4. Detection of food irradiation with luminescence methods

    International Nuclear Information System (INIS)

    Anderle, H.

    1997-06-01

    Food irradiation is applied as method for the preservation of foods, the prevention of food spoilage and the inhibition of food-borne pathogens. Doses exceeding 10 kGy (10 kJ/kg) are not recommended by the WHO. The different legislation requires methods for the detection and the closimetry of irradiated foods. Among the physical methods based on the radiation-induced changes in inorganic, nonhygroscopic crystalline solids are thermoluminescence (TL), photostimulated luminescence (PSL) and lyoluminescence (LL) measurement. The luminescence methods were tested on natural minerals. Pure quartz, feldspars, calcite, aragonite and dolomite of known origin were irradiated, read out and analyzed to determine the influence of luminescence-activators and deactivators. Carbonate minerals show an orange-red TL easily detectable by blue-sensitive photomultiplier tubes. TIL-inactive carbonate samples may be identified by a lyoluminescence method using the reaction of trapped irradiation-generated charge carriers with the solvent during crystal-lattice breakup. The fine-ground mineral is dissolved in an alkaline complexing agent/chemiluminescence sensitizer/chemiluminescence catalyst (EDTA/luminol/hemin) reagent mixture. The TL and PSL of quartz is too weak to contribute a significant part for the corresponding signals in polymineral dust. Alkali and soda feldspar show intense TL and PSL. The temperature maxima in the TL glow curves allow a clear distinction. PSL does not give this additional information, it suffers from bleaching by ambient light and requires light-protection. Grain disinfestated with low irradiation doses (500 Gy) may not identified by both TL and PSL measurement. The natural TL of feldspar particles may be overlap with the irradiation-induced TL of other minerals. As a routine method, irradiated spices are identified with TL measurement. The dust particles have to be enriched by heavy-liquid flotation and centrifugation. The PSL method allows a clear

  5. Sample Handling and Chemical Kinetics in an Acoustically Levitated Drop Microreactor

    Science.gov (United States)

    2009-01-01

    Accurate measurement of enzyme kinetics is an essential part of understanding the mechanisms of biochemical reactions. The typical means of studying such systems use stirred cuvettes, stopped-flow apparatus, microfluidic systems, or other small sample containers. These methods may prove to be problematic if reactants or products adsorb to or react with the container’s surface. As an alternative approach, we have developed an acoustically-levitated drop reactor eventually intended to study enzyme-catalyzed reaction kinetics related to free radical and oxidative stress chemistry. Microliter-scale droplet generation, reactant introduction, maintenance, and fluid removal are all important aspects in conducting reactions in a levitated drop. A three capillary bundle system has been developed to address these needs. We report kinetic measurements for both luminol chemiluminescence and the reaction of pyruvate with nicotinamide adenine dinucleotide, catalyzed by lactate dehydrogenase, to demonstrate the feasibility of using a levitated drop in conjunction with the developed capillary sample handling system as a microreactor. PMID:19769373

  6. Ultra-sensitive chemiluminescent detection of Staphylococcus aureus based on competitive binding of Staphylococcus protein A-modified magnetic beads to immunoglobulin G

    International Nuclear Information System (INIS)

    Xiong, Jie; Wang, Wenwen; Zhou, Yali; Kong, Weijun; Wang, Zhenxing; Fu, Zhifeng

    2016-01-01

    Staphylococcus protein A (SPA) is a surface protein only expressed naturally in the cell walls of Staphylococcus aureus (S. aureus) and binds specifically to the Fc region of immunoglobulin G (IgG). This fact can be utilized for the detection of S. aureus. Specifically, SPA-modified magnetic beads, compete with S. aureus pathogens for binding to rabbit IgG that previously was labeled with horseradish peroxidase (HRP). The beads were then magnetically separated, and chemiluminescence (CL) was generated by adding the reagents luminol and H_2O_2. Under optimal conditions, the intensity of CL decreases with increasing concentration of S. aureus over a very wide linear range (10 to 1.0 × 10"9 cfu·mL"−"1), with a limit of detection of 6.0 cfu·mL"−"1 at an S/N ratio of 3. The assay (including binding reaction, magnetic separation, washing of beads and detection) is completed within 50 min which is faster than many reported methods. It can well distinguish S. aureus from other Gram-positive and Gram-negative bacteria. The magnetic beads have the beneficial effect of eliminating undesired matrix effects and of concentrating the sample. The method was applied to the analysis of urine, apple juice and glucose injection samples spiked with S. aureus, and recoveries ranged from 85 to 107 %. (author)

  7. Glucose impairs aspirin inhibition in platelets through a NAD(P)H oxidase signaling pathway.

    Science.gov (United States)

    Kobzar, Gennadi; Mardla, Vilja; Samel, Nigulas

    2017-07-01

    Hyperglycemia has been suggested to play a role in the increased platelet resistance to antiplatelet therapy in patients with diabetes mellitus. Exposure to high glucose impairs platelet inhibition by aspirin. It has been found that antioxidant agents reduce the effect of glucose, confirming the involvement of reactive oxygen species (ROS) in the effect of glucose. The aim of the study was to examine the mechanism of ROS increase by high glucose in aspirin-treated platelets. Platelet aggregation was measured by the optical method, and the production of ROS was detected using luminol-dependent horseradish peroxidase-enhanced chemiluminescence. We found that glucose did not affect ADP-induced platelet aggregation. However, it reduced the effect of aspirin on platelet aggregation, which was accompanied by an increase in ROS generation. The inhibition of NAD(P)H oxidase (NOX) prevented the glucose effect and ROS generation. The same result was recorded after the inhibition of p38 mitogen-activated protein kinases (p38 MAPK), phospholipase A 2 (PLA 2 ) or 12-lipoxygenase (12-LOX). The inhibition of TxA 2 receptor did not decrease the effect of glucose indicating that the effect was not caused by activation of TxA 2 receptors. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Assessment of Antioxidant Activity of Spray Dried Extracts of Psidium guajava Leaves by DPPH and Chemiluminescence Inhibition in Human Neutrophils

    Directory of Open Access Journals (Sweden)

    M. R. V. Fernandes

    2014-01-01

    Full Text Available This work evaluated the physicochemical properties and antioxidant activity of spray dried extracts (SDE from Psidium guajava L. leaves. Different drying carriers, namely, maltodextrin, colloidal silicon dioxide, Arabic gum, and β-cyclodextrin at concentrations of 40 and 80% relative to solids content, were added to drying composition. SDE were characterized through determination of the total phenolic, tannins, and flavonoid content. Antioxidant potential of the SDE was assessed by two assays: cellular test that measures the luminol-enhanced chemiluminescence (LumCL produced by neutrophils stimulated with phorbol myristate acetate (PMA and the DPPH radical scavenging (DPPH* method. In both assays the antioxidant activity of the SDE occurred in a concentration-dependent manner and showed no toxicity to the cells. Using the CLlum method, the IC50 ranged from 5.42 to 6.50 µg/mL. The IC50 of the SDE ranged from 7.96 to 8.11 µg/mL using the DPPH• method. Psidium guajava SDE presented significant antioxidant activity; thus they show high potential as an active phytopharmaceutical ingredient. Our findings in human neutrophils are pharmacologically relevant since they indicate that P. guajava SDE is a potential antioxidant and anti-inflammatory agent in human cells.

  9. A sensitive chemiluminescent immunoassay to detect Chromotrope FB (Chr FB) in foods.

    Science.gov (United States)

    Xu, Kun; Long, Hao; Xing, Rongge; Yin, Yongmei; Eremin, Sergei A; Meng, Meng; Xi, Rimo

    2017-03-01

    Chromotrope FB (Chr FB) is a synthetic azo dye permitted for use in foods and medicines. An acceptable daily intake (ADI) of Chr FB was 0-0.5mg/kg in China. In this study, we synthesized a Chr FB hapten with an amino group to prepare its artificial immunogen. Polyclonal antibodies obtained from New Zealand rabbits were applied to develop an indirect competitive chemiluminescent immunoassay (icCLIA) to detect Chr FB in foods. A horseradish peroxidase (HRP)-luminol-H 2 O 2 system was used to yield CL signal with p-iodophenol as an enhancement reagent. The method showed good specificity towards Chr FB and could detect as low as 0.02ngmL -1 Chr FB in buffer, 0.07ngg -1 in yoghurt candy, 0.07ngg -1 in vitamin drink and 0.13ngg -1 in bread. Compared with HPLC method, the proposed method is more sensitive by two orders of magnitude. The accuracy and precision of this method are acceptable and comparable with HPLC method. Therefore, the proposed method could be used for rapid screening of Chr FB in the mentioned foodstuffs. Copyright © 2016. Published by Elsevier B.V.

  10. An automatic enzyme immunoassay based on a chemiluminescent lateral flow immunosensor.

    Science.gov (United States)

    Joung, Hyou-Arm; Oh, Young Kyoung; Kim, Min-Gon

    2014-03-15

    Microfluidic integrated enzyme immunosorbent assay (EIA) sensors are efficient systems for point-of-care testing (POCT). However, such systems are not only relatively expensive but also require a complicated manufacturing process. Therefore, additional fluidic control systems are required for the implementation of EIAs in a lateral flow immunosensor (LFI) strip sensor. In this study, we describe a novel LFI for EIA, the use of which does not require additional steps such as mechanical fluidic control, washing, or injecting. The key concept relies on a delayed-release effect of chemiluminescence substrates (luminol enhancer and hydrogen peroxide generator) by an asymmetric polysulfone membrane (ASPM). When the ASPM was placed between the nitrocellulose (NC) membrane and the substrate pad, substrates encapsulated in the substrate pad were released after 5.3 ± 0.3 min. Using this delayed-release effect, we designed and implemented the chemiluminescent LFI-based automatic EIA system, which sequentially performed the immunoreaction, pH change, substrate release, hydrogen peroxide generation, and chemiluminescent reaction with only 1 sample injection. In a model study, implementation of the sensor was validated by measuring the high sensitivity C-reactive protein (hs-CRP) level in human serum. © 2013 Elsevier B.V. All rights reserved.

  11. A Highly Sensitive Chemiluminometric Assay for Real-Time Detection of Biological Hydrogen Peroxide Formation.

    Science.gov (United States)

    Zhu, Hong; Jia, Zhenquan; Trush, Michael A; Li, Y Robert

    2016-05-01

    Hydrogen peroxide (H 2 O 2 ) is a major reactive oxygen species (ROS) produced by various cellular sources, especially mitochondria. At high levels, H 2 O 2 causes oxidative stress, leading to cell injury, whereas at low concentrations, this ROS acts as an important second messenger to participate in cellular redox signaling. Detection and measurement of the levels or rates of production of cellular H 2 O 2 are instrumental in studying the biological effects of this major ROS. While a number of assays have been developed over the past decades for detecting and/or quantifying biological H 2 O 2 formation, none has been shown to be perfect. Perhaps there is no perfect assay for sensitively and accurately quantifying H 2 O 2 as well as other ROS in cells, wherein numerous potential reactants are present to interfere with the reliable measurement of the specific ROS. In this context, each assay has its own advantages and intrinsic limitations. This article describes a highly sensitive assay for real-time detection of H 2 O 2 formation in cultured cells and isolated mitochondria. This assay is based on the luminol/horseradish peroxidase-dependent chemiluminescence that is inhibitable by catalase. The article discusses the usefulness and shortcomings of this chemiluminometric assay in detecting biological H 2 O 2 formation induced by beta-lapachone redox cycling with both cells and isolated mitochondria.

  12. Phenylboronic acid immunoaffinity reactor coupled with flow injection chemiluminescence for determination of {alpha}-fetoprotein

    Energy Technology Data Exchange (ETDEWEB)

    Wu Yafeng [Jiangsu Provincial Key Lab of Biomaterials and Biodevices, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 210096 (China); Zhuang Yafeng [Department of Chemistry, Changzhou Institute of Technology, Changzhou 213022 (China); Liu Songqin [Jiangsu Provincial Key Lab of Biomaterials and Biodevices, School of Chemistry and Chemical Engineering, Southeast University, Nanjing 210096 (China)], E-mail: liusq@seu.edu.cn; He Lin [Department of Chemistry, North Carolina State University, Raleigh, NC 27695-8204 (United States)

    2008-12-23

    A reusable and sensitive immunoassay based on phenylboronic acid immunoaffinity reactor in combination with flow injection chemiluminescence (CL) for determination of glycoprotein was described. The reactor was fabricated by immobilizing 3-aminophenylboronic acid (APBA) on glass microbeads with {gamma}-glycidoxypropyltrimethoxysilane (GPMS) as linkage. The {alpha}-fetoprotein (AFP) could be easily immobilized on the APBA coated beads through sugar-boronic interaction. After an off-line incubation, the mixture of the analyte AFP with horseradish peroxidase-labeled AFP antibody (HRP-anti-AFP) was injected into the reactor. This led the trapping of free HRP-anti-AFP by the surface coated AFP on glass beads. The trapped HRP-anti-AFP was detected by chemiluminescence due to its sensitizing effect on the reaction of luminol and hydrogen peroxide. Under optimal conditions, the chemiluminescent signal was proportional to AFP concentration in the range of 10-100 ng mL{sup -1}. The whole assay process including regeneration of the reactor could be completed within 31 min. The proposed system showed acceptable detection and fabrication reproducibility, and the results obtained with the present method were in acceptable agreement with those from parallel single-analyte test of practical clinical sera. The described method enabled a low-cost, time saving and was potential to detect the serum AFP level in clinical diagnosis.

  13. In situ-synthesized cadmium sulfide nanowire photosensor with a parylene passivation layer for chemiluminescent immunoassays.

    Science.gov (United States)

    Im, Ju-Hee; Kim, Hong-Rae; An, Byoung-Gi; Chang, Young Wook; Kang, Min-Jung; Lee, Tae-Geol; Son, Jin Gyeng; Park, Jae-Gwan; Pyun, Jae-Chul

    2017-06-15

    The direct in situ synthesis of cadmium sulfide (CdS) nanowires (NWs) was presented by direct synthesis of CdS NWs on the gold surface of an interdigitated electrode (IDE). In this work, we investigated the effect of a strong oxidant on the surfaces of the CdS NWs using X-ray photoelectron spectroscopy, transmission electron microscopy, and time-of-flight secondary ion mass spectrometry. We also fabricated a parylene-C film as a surface passivation layer for in situ-synthesized CdS NW photosensors and investigated the influence of the parylene-C passivation layer on the photoresponse during the coating of parylene-C under vacuum using a quartz crystal microbalance and a photoanalyzer. Finally, we used the in situ-synthesized CdS NW photosensor with the parylene-C passivation layer to detect the chemiluminescence of horseradish peroxidase and luminol and applied it to medical detection of carcinoembryonic antigen. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. An in-fiber integrated optofluidic device based on an optical fiber with an inner core.

    Science.gov (United States)

    Yang, Xinghua; Yuan, Tingting; Teng, Pingping; Kong, Depeng; Liu, Chunlan; Li, Entao; Zhao, Enming; Tong, Chengguo; Yuan, Libo

    2014-06-21

    A new kind of optofluidic in-fiber integrated device based on a specially designed hollow optical fiber with an inner core is designed. The inlets and outlets are built by etching the surface of the optical fiber without damaging the inner core. A reaction region between the end of the fiber and a solid point obtained after melting is constructed. By injecting samples into the fiber, the liquids can form steady microflows and react in the region. Simultaneously, the emission from the chemiluminescence reaction can be detected from the remote end of the optical fiber through evanescent field coupling. The concentration of ascorbic acid (AA or vitamin C, Vc) is determined by the emission intensity of the reaction of Vc, H2O2, luminol, and K3Fe(CN)6 in the optical fiber. A linear sensing range of 0.1-3.0 mmol L(-1) for Vc is obtained. The emission intensity can be determined within 2 s at a total flow rate of 150 μL min(-1). Significantly, this work presents information for the in-fiber integrated optofluidic devices without spatial optical coupling.

  15. The effect of lipid peroxidation products on reactive oxygen species formation and nitric oxide production in lipopolysaccharide-stimulated RAW 264.7 macrophages.

    Science.gov (United States)

    Ambrozova, Gabriela; Pekarova, Michaela; Lojek, Antonin

    2011-02-01

    Lipid peroxidation induced by oxidants leads to the formation of highly reactive metabolites. These can affect various immune functions, including reactive oxygen species (ROS) and nitric oxide (NO) production. The aim of the present study was to investigate the effects of lipid peroxidation products (LPPs) - acrolein, 4-hydroxynonenal, and malondialdehyde - on ROS and NO production in RAW 264.7 macrophages and to compare these effects with the cytotoxic properties of LPPs. Macrophages were stimulated with lipopolysaccharide (0.1 μg/ml) and treated with selected LPPs (concentration range: 0.1-100 μM). ATP test, luminol-enhanced chemiluminescence, Griess reaction, Western blotting analysis, amperometric and total peroxyl radical-trapping antioxidant parameter assay were used for determining the LPPs cytotoxicity, ROS and NO production, inducible nitric oxide synthase expression, NO scavenging, and antioxidant properties of LPPs, respectively. Our study shows that the cytotoxic action of acrolein and 4-hydroxynonenal works in a dose- and time-dependent manner. Further, our results imply that acrolein, 4-hydroxynonenal, and malondialdehyde can inhibit, to a different degree, ROS and NO production in stimulated macrophages, partially independently of their toxic effect. Also, changes in enzymatic pathways (especially NADPH-oxidase and nitric oxide synthase inhibition) and NO scavenging properties are included in the downregulation of reactive species formation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  16. Application of chemiluminescence to the study of alpha, beta and gamma radiolysis of water

    International Nuclear Information System (INIS)

    Broudic, V.; Muzeau, B.; Jegou, C.; Bonnal, M.; Gavazzi, A.; Marques, C.

    2004-01-01

    In the frame of the French research program on the long-term behavior of spent nuclear fuel, experiments are conducted in ATALANTE to develop and validate models of spent fuel evolution in contact with an aqueous phase. One of the mechanisms that may govern intermediate or long-term alteration of the spent fuel matrix in a repository is the oxidizing dissolution by radiolysis products of water. Leaching experiments in de-aerated media requires the analysis of hydrogen peroxide, as a major product of water radiolysis, down to 10 -8 mol.L -1 . This work presents the results obtained using the chemiluminescence reaction of iso-luminol with H 2 O 2 , catalyzed by micro-peroxidase. Depending on the samples used, different types of radiolytic processes were studied: α radiolysis of water when leaching UO 2 pellets doped with alpha emitters, or γ radiolysis of water when leaching the same samples or spent fuel in a gamma field. Influences of operating conditions on the analytical results are discussed. (authors)

  17. Aluminum induces lipid peroxidation and aggregation of human blood platelets

    Directory of Open Access Journals (Sweden)

    T.J.C. Neiva

    1997-05-01

    Full Text Available Aluminum (Al3+ intoxication is thought to play a major role in the development of Alzheimer's disease and in certain pathologic manifestations arising from long-term hemodialysis. Although the metal does not present redox capacity, it can stimulate tissue lipid peroxidation in animal models. Furthermore, in vitro studies have revealed that the fluoroaluminate complex induces diacylglycerol formation, 43-kDa protein phosphorylation and aggregation. Based on these observations, we postulated that Al3+-induced blood platelet aggregation was mediated by lipid peroxidation. Using chemiluminescence (CL of luminol as an index of total lipid peroxidation capacity, we established a correlation between lipid peroxidation capacity and platelet aggregation. Al3+ (20-100 µM stimulated CL production by human blood platelets as well as their aggregation. Incubation of the platelets with the antioxidants nor-dihydroguaiaretic acid (NDGA (100 µM and n-propyl gallate (NPG (100 µM, inhibitors of the lipoxygenase pathway, completely prevented CL and platelet aggregation. Acetyl salicylic acid (ASA (100 µM, an inhibitor of the cyclooxygenase pathway, was a weaker inhibitor of both events. These findings suggest that Al3+ stimulates lipid peroxidation and the lipoxygenase pathway in human blood platelets thereby causing their aggregation

  18. Free radical scavenging activity of coenzyme Q measured by a chemiluminescent assay

    International Nuclear Information System (INIS)

    Battino, Maurizio; Ferri, Elida; Girotti, Stefano; Lenaz, Giorgio

    1991-01-01

    Involvement of coenzyme Q (CoQ) in anti-oxydant activities, in addition to its major redox role, has frequently been suggested in recent years. In order to elucidate if CoQ could really be engaged in scavenging free radicals produced endogenously in a biological system, an experimental system was developed in which beef heart mitochondria in the presence of a saturating NADH concentration and of rotenone produce free radicals. The presence of oxygen-reactive forms was easily detected by a luminol-dependent chemiluminescence process. The chemi-luminescence assay showed that short-chain CoQ homologues can act as pro-oxidants, enhancing free radical effects, while exogenous coenzyme Q 10 could scavenge free radicals, especially at very low concentration. In this system, exogenous CoQ 10 was more effective than α-tocopherol at the same concentration in scavenging free radicals. The molecular mechanism that leads to this activity is still unclear, but these results are of biochemical importance because they indicate that CoQ may act as an anti=oxidant in situations mimicking physiopathological conditions. This direct chemiluminescent method is promising for studies of biochemical processes which involve active oxygen species. (author). 24 refs.; 4 figs

  19. Evaluation of Antibacterial Enrofloxacin in Eggs by Matrix Solid Phase Dispersion-Flow Injection Chemiluminescence

    Directory of Open Access Journals (Sweden)

    Xiaocui Duan

    2014-01-01

    Full Text Available The study based on the chemiluminescence (CL reaction of potassium ferricyanide and luminol in sodium hydroxide medium, enrofloxacin (ENRO could dramatically enhance CL intensities and incorporated with matrix solid-phase dispersion (MSPD technique (Florisil used as dispersant, dichloromethane eluted the target compounds. A simple flow injection chemiluminescence (FL-CL method with MSPD technique for determination of ENRO in eggs was described. Under optimal conditions, the CL intensities were linearly related to ENRO concentration ranging from 4.0×10-8 g.L−1 to 5.0×10-5 g.L−1, with a correlation coefficient of 0.9989 and detection limit of 5.0×10-9 g.L−1. The relative standard deviation was 3.6% at an ENRO concentration of 2.0×10-6 g.L−1. Our testing technique can help ensure food safety, and thus, protect public health.

  20. Determination of aminoglycoside antibiotics using an on-chip microfluidic device with chemiluminescence detection

    International Nuclear Information System (INIS)

    Sierra-Rodero, M.; Fernandez-Romero, J.M.; Gomez-Hens, A.

    2012-01-01

    We describe an on-chip microflow injection (μFI) approach for the determination of aminoglycoside antibiotics using chemiluminescence (CL) detection. The method is based on the inhibition of the Cu(II)-catalyzed CL reaction of luminol and hydrogen peroxide by the aminoglycosides due to the formation of a complex between the antibiotic and Cu(II). The main features of the method include small sample volumes and a fast response. Syringe pumps were used to insert the sample and the reagents into the microfluidic device. CL was collected using a fiber optic bundle connected to a luminescence detector. All instrumental, hydrodynamic and chemical variables involved in the system were optimized using neomycin as the aminoglycoside model. Inhibition is proportional to the concentration of the antibiotics. The dynamic ranges of the calibration graphs obtained for neomycin, streptomycin and amikacin are 0.3-3.3, 0.9-13.7, and 0.8-8.5 μmol L -1 , and the detection limits are 0.09, 0.28 and 0.24 μmol L -1 , respectively. The precision of the methods, expressed as relative standard deviation, is in the range from 0.8 to 5.0 %. The method was successfully applied to the determination of neomycin in water samples, with recoveries ranging from 80 to 120 %. (author)

  1. Effects of heme-PrP complex on cell-free conversion and peroxidase-linked immunodetection of prions in blood-based assays.

    Science.gov (United States)

    Soutyrine, Andrei; Yogasingam, Nishandan; Huang, Hongsheng; Mitchell, Gordon

    2015-08-01

    Prion protein (PrP) binding to natural and synthetic porphyrins has been previously demonstrated but the effects of endogenous heme interactions with PrP remain uncertain. This study investigated implications of this interaction in blood-based peroxidase-linked prion immunodetection and seeded conversion of cellular prion (PrP(C)) into disease associated form (PrP(Sc)). Heme binding to recombinant PrP(C) enhanced intrinsic peroxidase activity (POD) by 2.5-fold and POD inherent to denatured blood accounted for over 84% of luminol-based substrate oxidation in a prion immunodetection assay. An immuno-capture assay showed that 75-98% of blood POD was attributable to binding of PrP(C) with endogenous heme. Additionally, 10 μM heme inhibited (PPrP(C) to PrP(Sc) through the protein misfolding cycling amplification assay. We conclude that the observed effects can interfere with cell-free conversion and peroxidase-linked immunodetection of prions in blood-based assays. These results indicate that heme-PrP interactions could modulate intrinsic POD and protect PrP(C) from conversion into PrP(Sc). Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.

  2. Sonochemical cleaning efficiencies in dental instruments

    Science.gov (United States)

    Tiong, T. Joyce; Walmsley, A. Damien; Price, Gareth J.

    2012-05-01

    Ultrasound has been widely used for cleaning purposes in a variety of situations, including in dental practice. Cleaning is achieved through a combination of acoustically driven streaming effects and sonochemical effects arising from the production of inertial cavitation in a liquid. In our work, various dental instruments used for endodontic (root canal) treatment have been evaluated for their efficiency in producing sonochemical effects in an in-vitro cleaning environment. The areas where cavitation was produced were mapped by monitoring chemiluminescence from luminol solutions and this was correlated with their cleaning efficiencies - assessed by the ability to bleach a dye, to form an emulsion by mixing immiscible components and also to remove ink from a glass surface. The results showed good correlation (Pearson's coefficient > 0.9) between the cavitation and cleaning efficiencies, suggesting that the former plays an important role in cleaning. The methods developed and the results will be beneficial in endodontics research in order to optimise future root canal instruments and treatments.

  3. Correlation Between Metabolic Syndrome, Periodontitis and Reactive Oxygen Species Production. A Pilot Study.

    Science.gov (United States)

    Patini, Romeo; Gallenzi, Patrizia; Spagnuolo, Gianrico; Cordaro, Massimo; Cantiani, Monica; Amalfitano, Adriana; Arcovito, Alessandro; Callà, Cinzia; Mingrone, Gertrude; Nocca, Giuseppina

    2017-01-01

    Metabolic syndrome (MetS) is associated with an increased risk of periodontitis even if the mechanism is unknown. Since both MetS and periodontitis are characterized by an alteration of inflammation status, the aim of this pilot study was to determine if differences in ROS metabolism of phagocytes isolated from (A) patients with MetS, (B) patients with both MetS and mild periodontitis, (C) healthy subjects and (D) normal weight subjects with mild periodontitis, were present. ROS metabolism was studied by a Chemiluminescence (CL) technique: the system was made up of luminol (100 nmol/L) and cells (1 × 10 5 ) in the presence or absence of stimulus constituted by opsonized zymosan (0.5 mg). The final volume (1.0 mL) was obtained using modified KRP buffer. ROS production was measured at 25°C for 2 h, using an LB 953 luminometer (Berthold, EG & G Co, Germany). All the experiments were performed in triplicate. All results are mean ± standard deviation (SD). The group of means was compared by the analysis of variance "(ANOVA)". A value of p < 0.05 was considered significant. Results showed that basal ROS production (both from PMNs and from PBMs) of groups A, B and D was increased with respect to that obtained from group C ( p <0.05). These results are congruent with literature data, although the actual clinical relevance of the phenomenon remains to be evaluated.

  4. Sonochemical and high-speed optical characterization of cavitation generated by an ultrasonically oscillating dental file in root canal models.

    Science.gov (United States)

    Macedo, R G; Verhaagen, B; Fernandez Rivas, D; Gardeniers, J G E; van der Sluis, L W M; Wesselink, P R; Versluis, M

    2014-01-01

    Ultrasonically Activated Irrigation makes use of an ultrasonically oscillating file in order to improve the cleaning of the root canal during a root canal treatment. Cavitation has been associated with these oscillating files, but the nature and characteristics of the cavitating bubbles were not yet fully elucidated. Using sensitive equipment, the sonoluminescence (SL) and sonochemiluminescence (SCL) around these files have been measured in this study, showing that cavitation occurs even at very low power settings. Luminol photography and high-speed visualizations provided information on the spatial and temporal distribution of the cavitation bubbles. A large bubble cloud was observed at the tip of the files, but this was found not to contribute to SCL. Rather, smaller, individual bubbles observed at antinodes of the oscillating file with a smaller amplitude were leading to SCL. Confinements of the size of bovine and human root canals increased the amount of SL and SCL. The root canal models also showed the occurrence of air entrainment, resulting in the generation of stable bubbles, and of droplets, near the air-liquid interface and leading eventually to a loss of the liquid. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Antifatigue and Antioxidant Activity of Alcoholic Extract from Saussurea involucrata

    Directory of Open Access Journals (Sweden)

    Jang-Chang Lee

    2011-10-01

    Full Text Available Fatigue is a noticeable and highly prevalent symptom in tense, industriously, and economically affluent modern society. Therefore, new antifatigue agents to smooth the fatigue feature are an energetic topic. The total ethanol extract (ESI of Saussurea involucrata Kar et Kir., known as Tian-Shan snow lotus, was evaluated for antifatigue activity in ICR mice with mice forced swimming test and the determination of the contents of blood lactic acid and serum urea nitrogen. ESI (0.05, 0.15, 0.25 g/kg was administered orally to mice for 4 weeks. The average swimming times to exhaustion of the ESI-treated ICR mice (0.15, 0.25 g/kg were prolonged by 132% and 180% (p<0.001 with a lessening of fatigue compared with that of the control group. Analysis of biochemical parameters showed that levels of serum urea nitrogen and blood lactic acid of experimental groups were also decreased significantly (p<0.001 compared with that of the control group. The antioxidant activity of ESI was investigated by the 2,2-diphenyl-1-picrylhydrazyl (DPPH free radical-scavenging assay and the hydrogen peroxide-induced luminol chemiluminescence assay and the results indicated that ESI exerts DPPH scavenging ability and reducing power. These results provide scientific evidence that S. involucrata may have been potential as an antifatigue agent.

  6. Eco-friendly synthesis of gelatin-capped bimetallic Au-Ag nanoparticles for chemiluminescence detection of anticancer raloxifene hydrochloride.

    Science.gov (United States)

    Alarfaj, Nawal A; El-Tohamy, Maha F

    2016-09-01

    This study described the utility of green analytical chemistry in the synthesis of gelatin-capped silver, gold and bimetallic gold-silver nanoparticles (NPs). The preparation of nanoparticles was based on the reaction of silver nitrate or chlorauric acid with a 1.0 wt% aqueous gelatin solution at 50°C. The gelatin-capped silver, gold and bimetallic NPs were characterized using transmission electron microscopy, UV-vis, X-ray diffraction and Fourier transform infrared spectroscopy, and were used to enhance a sensitive sequential injection chemiluminescence luminol-potassium ferricyanide system for determination of the anticancer drug raloxifene hydrochloride. The developed method is eco-friendly and sensitive for chemiluminescence detection of the selected drug in its bulk powder, pharmaceutical injections and biosamples. After optimizing the conditions, a linear relationship in the range of 1.0 × 10(-9) to 1.0 × 10(-1)  mol/L was obtained with a limit of detection of 5.0 × 10(-10)  mol/L and a limit of quantification of 1.0 × 10(-9)  mol/L. Statistical treatment and method validation were performed based on ICH guidelines. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  7. A readout circuit dedicated for the detection of chemiluminescence using a silicon photomultiplier

    Science.gov (United States)

    Baszczyk, M.; Dorosz, P.; Mik, L.; Kucewicz, W.; Reczynski, W.; Sapor, M.

    2018-05-01

    A readout circuit dedicated for the detection of the chemiluminescence phenomenon using a silicon photomultiplier (SiPM) is presented. During chemiluminescence, light is generated as a result of chemical reaction. Chemiluminescence is used in many applications within medicine, chemistry, biology and biotechnology, and is one of the most important sensing techniques in biomedical science and clinical medicine. The front-end electronics consist of a preamplifier and a fast shaper—this produces pulses, the peaking time which is 3.6 ns for a single photon and the FWHM is 3.8 ns. The system has been optimised to measure chemiluminescence—it is sensitive at the level of single photons, it generates a low number of overlapping pulses and is accurate. Two methods of signal detection are analysed and compared: the counting of events and amplitude detection. The relationship between the chemiluminescence light intensity and the concentration of the chemical compound (luminol) is linear in the range of the tested concentrations and has strong linearity parameters and low prediction intervals.

  8. Química recreativa con agua oxigenada

    Directory of Open Access Journals (Sweden)

    Mª Luisa Aguilar Muñoz

    2011-01-01

    Full Text Available Con el objeto de despertar el interés por la química así como de tratar diversos conceptos químicos y biológicos, presentamos la reacción de descomposición catalítica del agua oxigenada (H2O2 en tres experiencias diferentes y con distintos catalizadores. Esta reacción es muy exotérmica, produciendo agua y oxígeno. En la primera experiencia, se mezclan en una probeta la disolución de agua oxigenada y un poco de lavavajillas. Al añadir yoduro de potasio, éste actúa como catalizador y aparece gran cantidad de espuma debido al oxígeno desprendido en la reacción. En la segunda, se utiliza como catalizador un óxido de manganeso finamente dividido, que al dejarlo caer en el recipiente que contiene agua oxigenada, produce un espectacular chorro de vapor de agua. En la última, se produce una llamativa reacción luminiscente entre el peróxido de hidrógeno y el luminol utilizando como catalizador el ión hierro contenido en el hexacianoferrato (III de potasio (ferricianuro de potasio.

  9. Investigation on a Sensitive Chemiluminescence System Based on Ni(IV Complex to Determine Two β2-Agonist Drugs in Urine and Swine Feed and Their Mechanism

    Directory of Open Access Journals (Sweden)

    Xiao Dong

    2018-04-01

    Full Text Available Veterinary drug residues, particularly traces of β2-agonists, can cause various kinds of harmful impact to the environment and public health. Here, a sensitive chemiluminescence (CL method incorporated with a flow injection analysis is developed for the determination of two β2-agonists [i.e., salbutamol (SAL and terbutaline (TEB]. The system is based on the CL reaction of Ni(IV complex with luminol in alkaline solutions, whereas SAL and TEB can significantly enhance CL intensities. Under optimum conditions, CL intensities are proportional to the SAL and TEB concentration in the range of 1.0 × 10−9 M to 5.0 × 10−7 M and 1.0 × 10−9 M to 1.0 × 10−7 M, respectively. The limits of detection (3σ are 1.0 × 10−11 M for TEB, and 1.3 × 10−11 for SAL respectively. Relative standard deviations (n = 11 are less than 2% for 5.0 × 10−8 M SAL and TEB. Possible reaction mechanisms for the CL system are suggested based on the CL system spectra, Ni(IV complex oxidation characteristics, and electron spin resonance (ESR techniques. The proposed method has been applied to the analysis of urine and swine feed samples with satisfactory results.

  10. Attempts to Image the Early Inflammatory Response during Infection with the Lymphatic Filarial Nematode Brugia pahangi in a Mouse Model.

    Directory of Open Access Journals (Sweden)

    Elmarie Myburgh

    Full Text Available Helminth parasites remain a major constraint upon human health and well-being in many parts of the world. Treatment of these infections relies upon a very small number of therapeutics, most of which were originally developed for use in animal health. A lack of high throughput screening systems, together with limitations of available animal models, has restricted the development of novel chemotherapeutics. This is particularly so for filarial nematodes, which are long-lived parasites with a complex cycle of development. In this paper, we describe attempts to visualise the immune response elicited by filarial parasites in infected mice using a non-invasive bioluminescence imaging reagent, luminol, our aim being to determine whether such a model could be developed to discriminate between live and dead worms for in vivo compound screening. We show that while imaging can detect the immune response elicited by early stages of infection with L3, it was unable to detect the presence of adult worms or, indeed, later stages of infection with L3, despite the presence of worms within the lymphatic system of infected animals. In the future, more specific reagents that detect secreted products of adult worms may be required for developing screens based upon live imaging of infected animals.

  11. Improvement on simultaneous determination of chromium species in aqueous solution by ion chromatography and chemiluminescence detection

    DEFF Research Database (Denmark)

    Gammelgaard, Bente; Liao, Y.P.; Jons, O.

    1997-01-01

    A sensitive method for the simultaneous determination of chromium(III) and chromium(VI) was chromatography and chemiluminescence detection. Two Dionex ion-exchange guard columns in series, CG5 and AG7, were used to separate chromium(III) from chromium(VI). Chromium(VI) was reduced by potassium......, the stabilities of reductant and luminol solutions were studied. The linear range of the calibration curve for chromium(III) and chromium(VI) was 1-400 mu g l(-1). The detection limit was 0.12 mu g l(-1) for chromium(III) and 0.09 mu g l(-1) for chromium(VI), respectively. The precision at the 20 mu g l(-1) level...... was 1.4% for chromium(III) and 2.5% for chromium(VI), respectively. The accuracy of the chromium(III) determination was determined by analysis of the NIST standard reference material 1643c, Trace elements in water with the result 19.1 +/- 1.0 mu g Cr(III) l(-1) (certified value 19.0 +/- 0.6 mu g Cr...

  12. Highly sensitive determination of diclofenac based on resin beads and a novel polyclonal antibody by using flow injection chemiluminescence competitive immunoassay

    Science.gov (United States)

    Shi, Jing; Xu, Mingxia; Tang, Qinghui; Zhao, Kang; Deng, Anping; Li, Jianguo

    2018-02-01

    A novel flow injection chemiluminescence immunoassay for simple, sensitive and low-cost detection of diclofenac was established based on specific binding of antigen and antibody. Carboxylic resin beads used as solid phase carrier materials provided good biocompatibility and large surface-to-volume ratio for modifying more coating antigen. There was a competitive process between the diclofenac in solution and the immobilized coating antigen to react with the limited binding sites of the polyclonal antibody to form the immunocomplex. The second antibody labelled with horseradish peroxidase was introduced into the immunosensor and trapped by captured polyclonal antibody against diclofenac, which could effectively amplify chemiluminescence signals of luminol-PIP-H2O2. Under optimal conditions, the diclofenac could be detected quantitatively. The chemiluminescence intensity decreased linearly with the logarithm of the diclofenac concentration in the range of 0.1-100 ng mL- 1 with a detection limit of 0.05 ng mL- 1 at a signal-to-noise ratio of 3. The immunosensor exhibited high sensitivity, specificity and acceptable stability. This easy-operated and cost-effective analytical method could be valuable for the diclofenac determination in real water samples.

  13. Pd@Au core-shell nanocrystals with concave cubic shapes: kinetically controlled synthesis and electrocatalytic properties.

    Science.gov (United States)

    Zhang, Ling; Niu, Wenxin; Zhao, Jianming; Zhu, Shuyun; Yuan, Yali; Hua, Lianzhe; Xu, Guobao

    2013-01-01

    A new type of concave cubic Pd@Au core-shell nanocrystals is synthesized through a kinetically controlled growth process. Pd nanocubes of 56 nm are used as the inner core, and CTAC and Br(-) are used as the capping agent and selective adsorbent, respectively. A suitable ratio of HAuCl4 and cubic Pd seeds and the presence of Br(-) anions are critical to the growth of the concave cubic Pd@Au core-shell nanocrystals. The fast deposition rate on the corners of the cubic Pd seeds promotes the overgrowth of the Au outer shell along the direction, leading to the formation of concave cubic nanostructures. The reduction process is monitored by the surface plasmon resonance spectra of the nanocrystals, and the extinction band became broader and red shifted as the nanocrystals became larger. The electrocatalytic properties of the concave cubic Pd@Au core-shell nanocrystals were investigated with the cathodic electrochemiluminescence reaction of luminol and H2O2. A possible electrocatalytic mechanism was proposed and analyzed.

  14. A multipumping flow system for in vitro screening of peroxynitrite scavengers.

    Science.gov (United States)

    Ribeiro, Marta F T; Dias, Ana C B; Santos, João L M; Fernandes, Eduarda; Lima, José L F C; Zagatto, Elias A G

    2007-09-01

    Peroxynitrite anion is a reactive nitrogen species formed in vivo by the rapid, controlled diffusion reaction between nitric oxide and superoxide radicals. By reacting with several biological molecules, peroxynitrite may cause important cellular and tissue deleterious effects, which have been associated with many diseases. In this work, an automated flow-based procedure for the in vitro generation of peroxynitrite and subsequent screening of the scavenging activity of selected compounds is developed. This procedure involves a multipumping flow system (MPFS) and exploits the ability of compounds such as lipoic acid, dihydrolipoic acid, cysteine, reduced glutathione, oxidized glutathione, sulindac, and sulindac sulfone to inhibit the chemiluminescent reaction of luminol with peroxynitrite under physiological simulated conditions. Peroxynitrite was generated in the MPFS by the online reaction of acidified hydrogen peroxide with nitrite, followed by a subsequent stabilization by merging with a sodium hydroxide solution to rapidly quench the developing reaction. The pulsed flow and the timed synchronized insertion of sample and reagent solutions provided by the MPFS ensure the establishment of the reaction zone only inside the flow cell, thus allowing maximum chemiluminescence emission detection. The results obtained for the assayed compounds show that, with the exception of oxidized glutathione, all are highly potent scavengers of peroxynitrite at the studied concentrations.

  15. Branched nanotrees with immobilized acetylcholine esterase for nanobiosensor applications

    Energy Technology Data Exchange (ETDEWEB)

    Risveden, Klas; Bhand, Sunil; Danielsson, Bengt [Department of Pure and Applied Biochemistry, Center for Chemistry and Chemical Engineering, Lund University, PO Box 124, SE-22100 Lund (Sweden); Dick, Kimberly A; Samuelson, Lars [Solid State Physics, Lund University, Box 118, S-22100 Lund (Sweden); Rydberg, Patrik, E-mail: Kimberly.Dick@ftf.lth.se [Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, University of Copenhagen, Universitetsparken 2, DK-2100 Copenhagen (Denmark)

    2010-02-05

    A novel lab-on-a-chip nanotree enzyme reactor is demonstrated for the detection of acetylcholine. The reactors are intended for use in the RISFET (regional ion sensitive field effect transistor) nanosensor, and are constructed from gold-tipped branched nanorod structures grown on SiN{sub x}-covered wafers. Two different reactors are shown: one with simple, one-dimensional nanorods and one with branched nanorod structures (nanotrees). Significantly higher enzymatic activity is found for the nanotree reactors than for the nanorod reactors, most likely due to the increased gold surface area and thereby higher enzyme binding capacity. A theoretical calculation is included to show how the enzyme kinetics and hence the sensitivity can be influenced and increased by the control of electrical fields in relation to the active sites of enzymes in an electronic biosensor. The possible effects of electrical fields employed in the RISFET on the function of acetylcholine esterase is investigated using quantum chemical methods, which show that the small electric field strengths used are unlikely to affect enzyme kinetics. Acetylcholine esterase activity is determined using choline oxidase and peroxidase by measuring the amount of choline formed using the chemiluminescent luminol reaction.

  16. Dual-signal amplification strategy for ultrasensitive chemiluminescence detection of PDGF-BB in capillary electrophoresis.

    Science.gov (United States)

    Cao, Jun-Tao; Wang, Hui; Ren, Shu-Wei; Chen, Yong-Hong; Liu, Yan-Ming

    2015-12-01

    Many efforts have been made toward the achievement of high sensitivity in capillary electrophoresis coupled with chemiluminescence detection (CE-CL). This work describes a novel dual-signal amplification strategy for highly specific and ultrasensitive CL detection of human platelet-derived growth factor-BB (PDGF-BB) using both aptamer and horseradish peroxidase (HRP) modified gold nanoparticles (HRP-AuNPs-aptamer) as nanoprobes in CE. Both AuNPs and HRP in the nanoprobes could amplify the CL signals in the luminol-H2 O2 CL system, owing to the excellent catalytic behavior of AuNPs and HRP in the CL system. Meanwhile, the high affinity of aptamer modified on the AuNPs allows detection with high specificity. As proof-of-concept, the proposed method was employed to quantify the concentration of PDGF-BB from 0.50 to 250 fm with a detection limit of 0.21 fm. The applicability of the assay was further demonstrated in the analysis of PDGF-BB in human serum samples with acceptable accuracy and reliability. The result of this study exhibits distinct advantages, such as high sensitivity, good specificity, simplicity, and very small sample consumption. The good performances of the proposed strategy provide a powerful avenue for ultrasensitive detection of rare proteins in biological sample, showing great promise in biochemical analysis. Copyright © 2015 John Wiley & Sons, Ltd.

  17. Development of a Capillary-driven, Microfluidic, Nucleic Acid Biosensor

    Directory of Open Access Journals (Sweden)

    Fei HE

    2011-12-01

    Full Text Available An ideal point-of-care device would incorporate the simplicity and reliability of a lateral flow assay with a microfluidic device. Our system consists of self-priming microfluidics with sealed conjugate pads of reagent delivery and an absorbent pad for additional fluid draw. Using poly (methyl methacrylate (PMMA as a substrate, we have developed a single-step surface modification method which allows strong capillary flow within a sealed microchannel. Conjugate pads within the device held trapped complex consisting of the magnetic beads and nucleic-acid-probe-conjugated horseradish peroxidase (HRP. Magnetic beads were released when sample entered the chamber and hybridized with the complex. The complex was immobilized over a magnet while a luminol co-reactant stream containing H2O2 was merged with the channel. A plate reader was able to quantify the chemiluminescence signal. This new format of biosensor will allow for a smaller and more sensitive biosensor, as well as commercial-scale manufacturing and low materials cost.

  18. Assessment of antioxidant activity of spray dried extracts of Psidium guajava leaves by DPPH and chemiluminescence inhibition in human neutrophils.

    Science.gov (United States)

    Fernandes, M R V; Azzolini, A E C S; Martinez, M L L; Souza, C R F; Lucisano-Valim, Y M; Oliveira, W P

    2014-01-01

    This work evaluated the physicochemical properties and antioxidant activity of spray dried extracts (SDE) from Psidium guajava L. leaves. Different drying carriers, namely, maltodextrin, colloidal silicon dioxide, Arabic gum, and β -cyclodextrin at concentrations of 40 and 80% relative to solids content, were added to drying composition. SDE were characterized through determination of the total phenolic, tannins, and flavonoid content. Antioxidant potential of the SDE was assessed by two assays: cellular test that measures the luminol-enhanced chemiluminescence (LumCL) produced by neutrophils stimulated with phorbol myristate acetate (PMA) and the DPPH radical scavenging (DPPH∗ method). In both assays the antioxidant activity of the SDE occurred in a concentration-dependent manner and showed no toxicity to the cells. Using the CLlum method, the IC50 ranged from 5.42 to 6.50 µg/mL. The IC50 of the SDE ranged from 7.96 to 8.11 µg/mL using the DPPH(•) method. Psidium guajava SDE presented significant antioxidant activity; thus they show high potential as an active phytopharmaceutical ingredient. Our findings in human neutrophils are pharmacologically relevant since they indicate that P. guajava SDE is a potential antioxidant and anti-inflammatory agent in human cells.

  19. Visible light assisted photodegradation of halocarbons on the dye modified TiO{sub 2} surface using visible light

    Energy Technology Data Exchange (ETDEWEB)

    Chatterjee, Debabrata; Dasgupta, Shimanti [Chemistry Section, Central Mechanical Engineering Research Institute, Durgapur 713209 (India); N Rao, Nageswar [Wastewater Technology Division, National Environmental Engineering Research Institute, Nagpur 400020 (India)

    2006-05-05

    Visible light assisted photodegradation of various halocarbons, viz. chloro-phenols, trichloroethylene, 1,2-dichloroethane and 1,4-dichlorobenzene, has been achieved on the surface of dye-modified TiO{sub 2} semiconductor. Thionine, eosinY, rhodamineB, methyleneblue, nileblueA and safranineO were used for the modification of TiO{sub 2} semiconductor photocatalyst. After 5h of irradiation with a 150W Xenon lamp, over 55-72% degradation of pollutants has been achieved. A working mechanism involving excitation of surface adsorbed dye, followed by charge injection into the TiO{sub 2} conduction band and formation of reactive {sup *}O{sub 2}{sup -}/{sup *}HO{sub 2} radicals, is proposed for the degradation of the enlisted organics to carbon dioxide. Efficacy of the photocatalytic system towards halocarbon degradation has been discussed in regard to excited state redox properties of the dyes selected for this study. Formation of {sup *}O{sub 2}{sup -} radical has been substantiated by using a chemiluminescent probe luminol. (author)

  20. Zinc oxide nanoparticle-enhanced ultrasensitive chemiluminescence immunoassay for the carcinoma embryonic antigen

    International Nuclear Information System (INIS)

    Pal, Souvik; Bhand, Sunil

    2015-01-01

    An ultrasensitive enzyme-linked immunosorbent assay (ELISA) is reported for the determination of carcinoma embryonic antigen (CEA) in human serum. It was realized using a microplate reader using a 384-well plate. Monoclonal antibody (Ab) against CEA (1° Ab) acting as the capture probe was immobilized on zinc oxide nanoparticles (ZnO-NPs) in the form of self-assembled monolayers (SAMs). CEA captured by 1° Ab was quantified using a sandwich ELISA wherein a polyclonal second antibody against CEA (2° Ab) was used for detection and quantified using an HRP-labeled secondary antibody (3° Ab). The ZnO-NPs-CEA capture probe was deposited on the bottom of the wells in order to enhance capture of CEA. A 3-fold enhancement in the chemiluminescence (CL) signal of luminol is found (compared to a conventional ELISA). CEA can be quantified by this method in concentrations as low as 1 pg · mL −1 . The upper limit of detection is 20 ng · mL −1 . The use of ZnO-NPs also imparts improved thermal stability. When stored at 4 °C in phosphate-buffered saline of pH 7.4, the probe displays stability of up to 30 days. (author)

  1. Measurement of hydroxyl radical production in ultrasonic aqueous solutions by a novel chemiluminescence method.

    Science.gov (United States)

    Hu, Yufei; Zhang, Zhujun; Yang, Chunyan

    2008-07-01

    Measurement methods for ultrasonic fields are important for reasons of safety. The investigation of an ultrasonic field can be performed by detecting the yield of hydroxyl radicals resulting from ultrasonic cavitations. In this paper, a novel method is introduced for detecting hydroxyl radicals by a chemiluminescence (CL) reaction of luminol-hydrogen peroxide (H2O2)-K5[Cu(HIO6)2](DPC). The yield of hydroxyl radicals is calculated directly by the relative CL intensity according to the corresponding concentration of H2O2. This proposed CL method makes it possible to perform an in-line and real-time assay of hydroxyl radicals in an ultrasonic aqueous solution. With flow injection (FI) technology, this novel CL reaction is sensitive enough to detect ultra trace amounts of H2O2 with a limit of detection (3sigma) of 4.1 x 10(-11) mol L(-1). The influences of ultrasonic output power and ultrasonic treatment time on the yield of hydroxyl radicals by an ultrasound generator were also studied. The results indicate that the amount of hydroxyl radicals increases with the increase of ultrasonic output power (< or = 15 W mL(-1)). There is a linear relationship between the time of ultrasonic treatment and the yield of H2O2. The ultrasonic field of an ultrasonic cleaning baths has been measured by calculating the yield of hydroxyl radicals.

  2. Design of a sensitive aptasensor based on magnetic microbeads-assisted strand displacement amplification and target recycling.

    Science.gov (United States)

    Li, Ying; Ji, Xiaoting; Song, Weiling; Guo, Yingshu

    2013-04-03

    A cross-circular amplification system for sensitive detection of adenosine triphosphate (ATP) in cancer cells was developed based on aptamer-target interaction, magnetic microbeads (MBs)-assisted strand displacement amplification and target recycling. Here we described a new recognition probe possessing two parts, the ATP aptamer and the extension part. The recognition probe was firstly immobilized on the surface of MBs and hybridized with its complementary sequence to form a duplex. When combined with ATP, the probe changed its conformation, revealing the extension part in single-strand form, which further served as a toehold for subsequent target recycling. The released complementary sequence of the probe acted as the catalyst of the MB-assisted strand displacement reaction. Incorporated with target recycling, a large amount of biotin-tagged MB complexes were formed to stimulate the generation of chemiluminescence (CL) signal in the presence of luminol and H2O2 by incorporating with streptavidin-HRP, reaching a detection limit of ATP as low as 6.1×10(-10)M. Moreover, sample assays of ATP in Ramos Burkitt's lymphoma B cells were performed, which confirmed the reliability and practicality of the protocol. Copyright © 2013 Elsevier B.V. All rights reserved.

  3. LC-DAD-MS (ESI+) analysis and antioxidant capacity of crocus sativus petal extracts.

    Science.gov (United States)

    Termentzi, Aikaterini; Kokkalou, Eugene

    2008-04-01

    In this study, various fractions isolated from the petals of Crocus sativus were assessed at first for their phenolic content both qualitatively and quantitatively and secondly for their antioxidant activity. The phytochemical analysis was carried out by LC-DAD-MS (ESI (+)) whereas the antioxidant potential was evaluated by applying two methodologies, the DPPH. radical scavenging activity test and the Co(II)-induced luminol chemiluminescence procedure. According to data obtained from these antioxidant tests, the diethyl ether, ethyl acetate and aqueous fractions demonstrated the strongest antioxidant capacity. Interestingly, the major constituents identified in these fractions correspond to kaempferol, quercetin, naringenin and some flavanone and flavanol derivatives glycosylated and esterified with phenylpropanoic acids. In addition, the presence of some nitrogen-containing substances, as well as other phenolics and phenylpropanoic derivatives was also traced. The identification and structural elucidation of all substances isolated in this study was achieved by both comparing available literature data and by proposed fragmentation mechanisms based on evaluating the LC-DAD-MS (ESI (+)) experimental data. The quantitative analysis data obtained thus far have shown that Crocus sativus petals are a rich source of flavonoids. Such a fact suggests that the good antioxidant capacity detected in the various fractions of Crocus sativus petals could be attributed to the presence of flavonoids, since it is already known that these molecules exert antioxidant capability. The latter, along with the use of Crocus sativus in food and pharmaceutical industry is discussed.

  4. Oxidative response of neutrophils to platelet-activating factor is altered during acute ruminal acidosis induced by oligofructose in heifers.

    Science.gov (United States)

    Concha, Claudia; Carretta, María Daniella; Alarcón, Pablo; Conejeros, Ivan; Gallardo, Diego; Hidalgo, Alejandra Isabel; Tadich, Nestor; Cáceres, Dante Daniel; Hidalgo, María Angélica; Burgos, Rafael Agustín

    2014-01-01

    Reactive oxygen species (ROS) production is one of the main mechanisms used to kill microbes during innate immune response. D-lactic acid, which is augmented during acute ruminal acidosis, reduces platelet activating factor (PAF)-induced ROS production and L-selectin shedding in bovine neutrophils in vitro. This study was conducted to investigate whether acute ruminal acidosis induced by acute oligofructose overload in heifers interferes with ROS production and L-selectin shedding in blood neutrophils. Blood neutrophils and plasma were obtained by jugular venipuncture, while ruminal samples were collected using rumenocentesis. Lactic acid from plasma and ruminal samples was measured by HPLC. PAF-induced ROS production and L-selectin shedding were measured in vitro in bovine neutrophils by a luminol chemiluminescence assay and flow cytometry, respectively. A significant increase in ruminal and plasma lactic acid was recorded in these animals. Specifically, a decrease in PAF-induced ROS production was observed 8 h after oligofructose overload, and this was sustained until 48 h post oligofructose overload. A reduction in PAF-induced L-selectin shedding was observed at 16 h and 32 h post oligofructose overload. Overall, the results indicated that neutrophil PAF responses were altered in heifers with ruminal acidosis, suggesting a potential dysfunction of the innate immune response.

  5. Enzymatic logic calculation systems based on solid-state electrochemiluminescence and molecularly imprinted polymer film electrodes.

    Science.gov (United States)

    Lian, Wenjing; Liang, Jiying; Shen, Li; Jin, Yue; Liu, Hongyun

    2018-02-15

    The molecularly imprinted polymer (MIP) films were electropolymerized on the surface of Au electrodes with luminol and pyrrole (PY) as the two monomers and ampicillin (AM) as the template molecule. The electrochemiluminescence (ECL) intensity peak of polyluminol (PL) of the AM-free MIP films at 0.7V vs Ag/AgCl could be greatly enhanced by AM rebinding. In addition, the ECL signals of the MIP films could also be enhanced by the addition of glucose oxidase (GOD)/glucose and/or ferrocenedicarboxylic acid (Fc(COOH) 2 ) in the testing solution. Moreover, Fc(COOH) 2 exhibited cyclic voltammetric (CV) response at the AM-free MIP film electrodes. Based on these results, a binary 3-input/6-output biomolecular logic gate system was established with AM, GOD and Fc(COOH) 2 as inputs and the ECL responses at different levels and CV signal as outputs. Some functional non-Boolean logic devices such as an encoder, a decoder and a demultiplexer were also constructed on the same platform. Particularly, on the basis of the same system, a ternary AND logic gate was established. The present work combined MIP film electrodes, the solid-state ECL, and the enzymatic reaction together, and various types of biomolecular logic circuits and devices were developed, which opened a novel avenue to construct more complicated bio-logic gate systems. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Interferences of commercial NO2 instruments in the urban atmosphere and in a smog chamber

    Directory of Open Access Journals (Sweden)

    J. Kleffmann

    2012-01-01

    Full Text Available Reliable measurements of atmospheric trace gases are necessary for both, a better understanding of the chemical processes occurring in the atmosphere, and for the validation of model predictions. Nitrogen dioxide (NO2 is a toxic gas and is thus a regulated air pollutant. Besides, it is of major importance for the oxidation capacity of the atmosphere and plays a pivotal role in the formation of ozone and acid precipitation. Detection of NO2 is a difficult task since many of the different commercial techniques used are affected by interferences. The chemiluminescence instruments that are used for indirect NO2 detection in monitoring networks and smog chambers use either molybdenum or photolytic converters and are affected by either positive (NOy or negative interferences (radical formation in the photolytic converter. Erroneous conclusions on NO2 can be drawn if these interferences are not taken into consideration. In the present study, NO2 measurements in the urban atmosphere, in a road traffic tunnel and in a smog-chamber using different commercial techniques, i.e. chemiluminescence instruments with molybdenum or photolytic converters, a Luminol based instrument and a new NO2-LOPAP, were compared with spectroscopic techniques, i.e. DOAS and FTIR. Interferences of the different instruments observed during atmospheric measurements were partly characterised in more detail in the smog chamber experiments. Whereas all the commercial instruments showed strong interferences, excellent agreement was obtained between a new NO2-LOPAP instrument and the FTIR technique for the measurements performed in the smog chamber.

  7. Heterogeneity of neutrophil antibodies in patients with primary Sjögren's syndrome.

    Science.gov (United States)

    Lamour, A; Le Corre, R; Pennec, Y L; Cartron, J; Youinou, P

    1995-11-01

    Our aims were to determine the prevalence of neutrophil antibodies in patients with primary Sjögren's syndrome (pSS), identify their target antigen(s), and evaluate their functional significance. Neutrophil antibodies were detected using an indirect immunofluorescence (IIIF) test and an enzyme-linked immunosorbent assay (ELISA), using recombinant human Fc-gamma receptor (Fc gamma RIIIb) as a capture agent. Luminol-dependent chemiluminescence was then measured by an established technique. Antibodies to neutrophils were detected in 30 of 66 patients (45%) and categorized on the basis of positivity for the two assays: IIF+/ELISA+ (group A: five patients), IIF+/ELISA- (group B: five patients), and IFF-/ELISA+ (group C: 20 patients). All positive sera contained antibodies directed to the neutrophil specific Fc gamma RIIIb, and none of them bound to NAnull neutrophils. The titer of neutrophil-reactive antibodies (groups A and B) showed no correlation with the neutrophil count, but these autoantibodies did reduce the cell ability to generate a respiratory burst. Thus, neutrophil antibodies are common in patients with pSS. Their main target appears to be Fc gamma RIII, and this may partly account for the dysfunction in Fc gamma R-mediated clearance by the reticuloendothelial system reported in these patients.

  8. DNA origami nanorobot fiber optic genosensor to TMV.

    Science.gov (United States)

    Torelli, Emanuela; Manzano, Marisa; Srivastava, Sachin K; Marks, Robert S

    2018-01-15

    In the quest of greater sensitivity and specificity of diagnostic systems, one continually searches for alternative DNA hybridization methods, enabling greater versatility and where possible field-enabled detection of target analytes. We present, herein, a hybrid molecular self-assembled scaffolded DNA origami entity, intimately immobilized via capture probes linked to aminopropyltriethoxysilane, onto a glass optical fiber end-face transducer, thus producing a novel biosensor. Immobilized DNA nanorobots with a switchable flap can then be actuated by a specific target DNA present in a sample, by exposing a hemin/G-quadruplex DNAzyme, which then catalyzes the generation of chemiluminescence, once the specific fiber probes are immersed in a luminol-based solution. Integrating organic nanorobots to inorganic fiber optics creates a hybrid system that we demonstrate as a proof-of-principle can be utilized in specific DNA sequence detection. This system has potential applications in a wide range of fields, including point-of-care diagnostics or cellular in vivo biosensing when using ultrathin fiber optic probes for research purposes. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Chemical Constituents with Free-Radical-Scavenging Activities from the Stem of Microcos paniculata

    Directory of Open Access Journals (Sweden)

    Yu Chen

    2010-08-01

    Full Text Available The free-radical-scavenging activities of various solvent extracts of Microcos paniculata were evaluated through in vitro model systems, such as 1,1-diphenyl-2-picrylhydrazyl (DPPH, 2,2'-azino-bis-(3-ethylbenzothiazoline-6-sulfonate (ABTS and Co (II EDTA-induced luminol chemiluminescence by flow injection. In all three of these systems the ethyl acetate (EtOAc extract showed the highest free-radical-scavenging activity compared with the other three (n-BuOH, water and petroleum ether extracts. Free-radical-scavenging assay-guided chromatographic separation of the EtOAc extract, using a normal-phase and reverse-phase silica gel column chromatography yielded five compounds: a new triterpene named methyl 3b-O-p-hydroxy-E-cinnamoyloxy-2a,23-dihydroxyolean-12-en-28-oate (1, whose spectral data are presented for the first time, together with four known compounds, epicatechin (2, 3-trans-feruloyl maslinic acid (3, maslinic acid (4 and sucrose (5. All of the compounds were isolated from Microcos paniculata for the first time. The compounds were identified by spectroscopic methods. Among them, compound 2 displayed significant free-radical-scavenging activity which is similar to that of standard antioxidant ascorbic acid (VC and therefore may be a promising natural antioxidant.

  10. Chemiluminescent Determination of Oxamyl in Drinking Water and Tomato Using Online Postcolumn UV Irradiation in a Chromatographic System.

    Science.gov (United States)

    Murillo Pulgarín, José A; García Bermejo, Luisa F; Durán, Armando Carrasquero

    2018-03-07

    High-performance liquid chromatography (HPLC) was used to separate oxamyl from other pesticides in drinking water and tomato paste. The eluate emerging from the column tail was mixed with an alkaline solution of Co 2+ in EDTA and irradiated with UV light to induce photolysis of the carbamate in order to obtain free radicals and other reactive species that oxidize luminol and produce chemiluminescence (CL) as a result. The intensity of the CL signal was monitored in the form of chromatographic peaks. Under the optimum operating conditions for the HPLC-UV-CL system, the analyte concentration was linearly related to peak area. The limit of detection as determined in accordance with the IUPAC criterion was 0.17 mg L -1 . Oxamyl was successfully extracted with recoveries of 88.7-103.1% from spiked tomato paste by using a simple QuEChERS (Quick, Easy, Cheap, Effective, Rugged, and Safe) sample preparation approach. Similar recoveries were obtained from drinking water samples spiked with oxamyl concentrations above the LOD. The proposed method is a simple, fast, accurate choice for quantifying this pesticide.

  11. Advantages and limitations of common testing methods for antioxidants.

    Science.gov (United States)

    Amorati, R; Valgimigli, L

    2015-05-01

    Owing to the importance of antioxidants in the protection of both natural and man-made materials, a large variety of testing methods have been proposed and applied. These include methods based on inhibited autoxidation studies, which are better followed by monitoring the kinetics of oxygen consumption or of the formation of hydroperoxides, the primary oxidation products. Analytical determination of secondary oxidation products (e.g. carbonyl compounds) has also been used. The majority of testing methods, however, do not involve substrate autoxidation. They are based on the competitive bleaching of a probe (e.g. ORAC assay, β-carotene, crocin bleaching assays, and luminol assay), on reaction with a different probe (e.g. spin-trapping and TOSC assay), or they are indirect methods based on the reduction of persistent radicals (e.g. galvinoxyl, DPPH and TEAC assays), or of inorganic oxidizing species (e.g. FRAP, CUPRAC and Folin-Ciocalteu assays). Yet other methods are specific for preventive antioxidants. The relevance, advantages, and limitations of these methods are critically discussed, with respect to their chemistry and the mechanisms of antioxidant activity. A variety of cell-based assays have also been proposed, to investigate the biological activity of antioxidants. Their importance and critical aspects are discussed, along with arguments for the selection of the appropriate testing methods according to the different needs.

  12. A magnetic particles-based chemiluminescence enzyme immunoassay for rapid detection of ovalbumin.

    Science.gov (United States)

    Feng, Xiao-Li; Ren, Hong-Lin; Li, Yan-Song; Hu, Pan; Zhou, Yu; Liu, Zeng-Shan; Yan, Dong-Ming; Hui, Qi; Liu, Dong; Lin, Chao; Liu, Nan-Nan; Liu, Yan-Yan; Lu, Shi-Ying

    2014-08-15

    Egg allergy is an important public health and safety concern, so quantification and administration of food or vaccines containing ovalbumin (OVA) are urgently needed. This study aimed to establish a rapid and sensitive magnetic particles-chemiluminescence enzyme immunoassay (MPs-CLEIA) for the determination of OVA. The proposed method was developed on the basis of a double antibodies sandwich immunoreaction and luminol-H2O2 chemiluminescence system. The MPs served as both the solid phase and separator, the anti-OVA MPs-coated polyclonal antibodies (pAbs) were used as capturing antibody, and the horseradish peroxidase (HRP)-labeled monoclonal antibody (mAb) was taken as detecting antibody. The parameters of the method were evaluated and optimized. The established MPs-CLEIA method had a linear range from 0.31 to 100ng/ml with a detection limit of 0.24ng/ml. The assays showed low reactivities and less than 5% of intraassay and interassay coefficients of variation (CVs), and the average recoveries were between 92 and 97%. Furthermore, the developed method was applied in real samples analysis successfully, and the correlation coefficient with the commercially available OVA kit was 0.9976. Moreover, it was more rapid and sensitive compared with the other methods for testing OVA. Copyright © 2014 Elsevier Inc. All rights reserved.

  13. Soil emissions of nitric oxide in a seasonally dry tropical forest of Mexico

    Science.gov (United States)

    Davidson, Eric A.; Vitousek, Peter M.; Riley, Ralph; Matson, Pamela A.; Garcia-Mendez, Georgina; Maass, J. M.

    1991-01-01

    Soil emissions of NO were measured at the Chamela Biological Station, Mexico, using soil covers and a field apparatus of NO detection based on CrO3 conversion of NO to NO2 and detection of NO2 by chemiluminescence with Luminol. Mean NO fluxes from forest soils ranged from 0.14 to 0.52 ng NO-N/sq cm/hr during the dry season and from 0.73 to 1.27 ng NO-N/sq cm/hr during the wet season. A fertilized floodplain pasture exhibited higher fluxes, but an unfertilized upland pasture, which represents the fastest growing land use in the region, had flux rates similar to the forest sites. Wetting experiments at the end of the dry season caused large pulses of NO flux, equaling 10 percent to 20 percent of the estimated annual NO emissions of 0.5-1.0 kg N/ha from the forest sites. Absence of a forest canopy during the dry season and the first wet season rain probably results in substantial NO(x) export from the forest system that may be important to regional atmospheric chemical processes. Wetting experiments during the wet season and a natural rain event had little or no stimulatory effect on NO flux rates.

  14. Minocycline affects human neutrophil respiratory burst and transendothelial migration.

    Science.gov (United States)

    Parenti, Astrid; Indorato, Boris; Paccosi, Sara

    2017-02-01

    This study aimed at investigating the in vitro activity of minocycline and doxycycline on human polymorphonuclear (h-PMN) cell function. h-PMNs were isolated from whole venous blood of healthy subjects; PMN oxidative burst was measured by monitoring ROS-induced oxidation of luminol and transendothelial migration was studied by measuring PMN migration through a monolayer of human umbilical vein endothelial cells. Differences between multiple groups were determined by ANOVA followed by Tukey's multiple comparison test; Student's t test for unpaired data for two groups. Minocycline (1-300 µM) concentration dependently and significantly inhibited oxidative burst of h-PMNs stimulated with 100 nM fMLP. Ten micromolar concentrations, which are superimposable to C max following a standard oral dose of minocycline, promoted a 29.8 ± 4 % inhibition of respiratory burst (P minocycline impaired PMN transendothelial migration, with maximal effect at 100 µM (42.5 ± 7 %, inhibition, n = 5, P minocycline exerted on innate immune h-PMN cell function.

  15. Chemiluminescent Diagnostics of Free-Radical Processes in an Abiotic System and in Liver Cells in the Presence of Nanoparticles Based on Rare-Earth Elements nReVO4:Eu3+ (Re = Gd, Y, La) and CeO2

    Science.gov (United States)

    Averchenko, E. A.; Kavok, N. S.; Klochkov, V. K.; Malyukin, Yu. V.

    2014-11-01

    We have used luminol-dependent chemiluminescence with Fenton's reagent to study the effect of nanoparticles based on rare-earth elements of different sizes and shapes on free-radical processes in abiotic and biotic cell-free systems, and also in isolated cells in vitro. We have estimated the effects of rare-earth orthovanadate nanoparticles of spherical (GdYVO4:Eu3+, 1-2 nm), spindle-shaped (GdVO4:Eu3+, 25 ×8 nm), and rod-shaped (LaVO4:Eu3+, 57 × (6-8) nm) nanoparticles and spherical CeO2 nanoparticles (sizes 1-2 nm and 8-10 nm). We have shown that in contrast to the abiotic system, in which all types of nanoparticles exhibit antiradical activity, in the presence of biological material, extra-small spherical (1-2 nm) nanoparticles of both types exhibit pro-oxidant activity, and also enhance pro-oxidant induced oxidative stress (for the pro-oxidants hydrogen peroxide and tert-butyl hydroperoxide). The effect of rare-earth orthovanadate spindle and rod shaped nanoparticles in this system was neutral; a moderate antioxidant effect was exhibited by 8-10 nm CeO2 nanoparticles.

  16. Antioxidant effects of cultured wild ginseng root extracts on the male reproductive function of boars and guinea pigs.

    Science.gov (United States)

    Yun, Suk Jun; Bae, Gui-Seck; Park, Jae Hawn; Song, Tae Ho; Choi, Ahreum; Ryu, Buom-Yong; Pang, Myung-Geol; Kim, Eun Joong; Yoon, Minjung; Chang, Moon Baek

    2016-07-01

    The main objective of this study was to investigate the effects of cultured wild ginseng root extracts (cWGRE) on the sperm of boars and the reproductive system of guinea pigs. Firstly, semen collected from boars (n=10) were incubated in 38°C for 1h with xanthine and xanthine oxidase to generate ROS. The cWGRE was added to the sperm culture system to test its antioxidant effect on the boar sperm. The amount of Reactive Oxygen Species (ROS) was measured by a chemiluminescence assay using luminol. The results indicated that the addition of cWGRE to boar sperm culture inhibited xanthine and xanthine oxidase-induced ROS concentrations. Treatment with cWGRE also had a positive effect on maintaining sperm motility. Effects of cWGRE administration on vitamin C-deficient guinea pigs were further investigated. Hartley guinea pigs (n=25) at 8 weeks of age were randomly divided into five groups. With the exception of the positive control group, each group was fed vitamin C-deficient feed for 21days (d). Respective groups were also orally administered cWGRE, ginseng extract, or mixed ginsenosides for 21 days. In comparison to the control group, oral administration of cWGRE reduced (Pproduction. Copyright © 2016 Elsevier B.V. All rights reserved.

  17. Enhanced Silver Nanoparticle Chemiluminescence Method for the Determination of Gemifloxacin Mesylate using Sequential Injection Analysis

    International Nuclear Information System (INIS)

    Alarfaj, N.A.; Aly, F.A.; Tamimi, A.A.

    2013-01-01

    A sequential injection analysis (SIA) with chemiluminescence detection has been proposed for the determination of the antibiotic gemifloxacin mesylate (GFX). The developed method is based on the enhancement effect of silver nanoparticles (Ag NPs) on the chemiluminescence (CL) signal of luminol-potassium ferricyanide reaction in alkaline medium. The introduction of gemifloxacin in this system produced a significant decrease in the CL intensity in presence of (Ag NPs). The optimum conditions for CL emission were investigated. Linear relationship between the decrease in CL intensity and concentration was obtained in the range 0.01-1000 ng mL-1, (r = 0.9997) with detection limit of 2.0 pg mL-1 and quantification limit of 0.01 pg mL-1. The relative standard deviation was 1.3 %. The proposed method was employed for the determination of gemifloxacin in bulk drug, in its pharmaceutical dosage forms and biological fluids such as human serum and urine. The interference of some common additive compounds such as glucose, lactose, starch, talc and magnesium stearate was investigated, and no interference was found from these excipients. The obtained SIA results were statistically compared with those obtained from a reported method and did not show any significant difference at confidence level 95%. (author)

  18. Antioxidant effects of herbal therapies used by patients with inflammatory bowel disease: an in vitro study.

    Science.gov (United States)

    Langmead, L; Dawson, C; Hawkins, C; Banna, N; Loo, S; Rampton, D S

    2002-02-01

    Herbal remedies used by patients for treatment of inflammatory bowel disease include slippery elm, fenugreek, devil's claw, Mexican yam, tormentil and wei tong ning, a traditional Chinese medicine. Reactive oxygen metabolites produced by inflamed colonic mucosa may be pathogenic. Aminosalicylates (5-ASA) are antioxidant and other such agents could be therapeutic. To assess the antioxidant effects of herbal remedies in cell-free oxidant-generating systems and inflamed human colorectal biopsies. Luminol-enhanced chemiluminescence in a xanthine/xanthine oxidase cell-free system was used to detect superoxide scavenging by herbs and 5-ASA, and fluorimetry to define peroxyl radical scavenging using a phycoerythrin degradation assay. Chemiluminescence was used to detect herbal effects on generation of oxygen radicals by mucosal biopsies from patients with active ulcerative colitis. Like 5-ASA, all herbs, except fenugreek, scavenged superoxide dose-dependently. All materials tested scavenged peroxyl dose-dependently. Oxygen radical release from biopsies was reduced after incubation in all herbs except Mexican yam, and by 5-ASA. All six herbal remedies have antioxidant effects. Fenugreek is not a superoxide scavenger, while Mexican yam did not inhibit radical generation by inflamed biopsies. Slippery elm, fenugreek, devil's claw, tormentil and wei tong ning merit formal evaluation as novel therapies in inflammatory bowel disease.

  19. (±)-2-Chloropropionic acid elevates reactive oxygen species formation in human neutrophil granulocytes

    International Nuclear Information System (INIS)

    Aam, B.B.; Fonnum, F.

    2006-01-01

    (±)-2-Chloropropionic acid (2-CPA) is a neurotoxic compound which kills cerebellar granule cells in vivo, and makes cerebellar granule cells in vitro produce reactive oxygen species (ROS). We have studied the effect of 2-CPA on ROS formation in human neutrophil granulocytes in vitro. We found an increased formation of ROS after 2-CPA exposure using three different methods; the fluorescent probe DCFH-DA and the chemiluminescent probes lucigenin and luminol. Four different inhibitors of ROS formation were tested on the cells in combination with 2-CPA to characterize the signalling pathways. The spin-trap s-PBN, the ERK1/2 inhibitor U0126 and the antioxidant Vitamin E inhibited the 2-CPA-induced ROS formation completely, while the mitochondrial transition permeability pore blocker cyclosporine A inhibited the ROS formation partly. We also found that 2-CPA induced an increased nitric oxide production in the cells by using the Griess reagent. The level of reduced glutathione, measured with the DTNB assay, was decreased after exposure to high concentrations of 2-CPA. Western blotting analysis showed that 2-CPA exposure led to an elevated phosphorylation of ERK MAP kinase. This phosphorylation was inhibited by U0126. Based on these experiments it seems like the mechanisms for 2-CPA induced toxicity involves ROS formation and is similar in neutrophil granulocytes as earlier shown in cerebellar granule cells. This also implies that 2-CPA may be immunotoxic

  20. Evaluation of Sonochemiluminescence in a Phantom in the Presence of Protoporphyrin IX Conjugated to Nanoparticles

    International Nuclear Information System (INIS)

    Shanei, A.; Sazgarnia, A.; Hassanzadeh-Kayyat, M.; Eshghi, H.; Soudmand, S.; Attaran Kakhki, N.

    2012-01-01

    When a liquid is irradiated with high-intensity and low-frequency ultrasound, acoustic cavitation occurs and there are some methods to determine and quantify this phenomenon. The existing methods for performing these experiments include sonochemiluminescence and chemical dosimetric methods. The particles in a liquid decrease the ultrasonic intensity threshold needed for cavitation onset. In this study, a new nano conjugate made up of Protoporphyrin IX and gold nanoparticles, i.e., Au-PpIX was used to provide nucleation sites for cavitation. The nonradiative relaxation time of PpIX in the presence of GNPs is longer than the similar time for PpIX without GNPs. This effect can be used in medical diagnostic and therapeutic applications. The acoustic cavitation activity was investigated studying integrated sonochemiluminescence signal in the wavelength range of 400-500 nm in polyacrylamide gel phantom containing luminol using a cooled CCD spectrometer at different intensities of 1 MHz ultrasound. In order to confirm these results, a chemical dosimetric method was utilized, too. sonochemiluminescence signal level in gel phantom containing Au-PpIX was higher than the other phantoms. These results have been confirmed by the chemical dosimetric data. This finding can be related to the existence of PpIX as a sensitizer and GNPs as cavitation nuclei. In other words, nanoparticles have acted as the sites for cavitation and have increased the cavitation rate. Another theory is that activation of PpIX has produced more free radicals and has enhanced the sonochemiluminescence signal level.

  1. Protective effect of low dose caffeine on psychological stress and cognitive function.

    Science.gov (United States)

    Çakır, Özgür Kasımay; Ellek, Nurfitnat; Salehin, Nabila; Hamamcı, Rabia; Keleş, Hülya; Kayalı, Damla Gökçeoğlu; Akakın, Dilek; Yüksel, Meral; Özbeyli, Dilek

    2017-01-01

    Caffeine is an adrenergic antagonist that enhances neuronal activity. Psychological stress depresses cognitive function. To investigate the effects of acute and chronic low dose caffeine on anxiety-like behavior and cognitive functions of acute or chronic psychological stressed rats. Acute or chronic caffeine (3mg/kg) was administered to male Sprague Dawley rats (200-250g, n=42) before acute (cat odor) and chronic variable psychological stress (restraint overcrowding stress, elevated plus maze, cat odor, forced swimming) induction. Anxiety and cognitive functions were evaluated by hole-board and object recognition tests. The brain glutathione and malondialdehyde assays, myeloperoxidase, nitric oxide (NO), superoxide dismutase (SOD), luminol and lucigenin activity and histological examination were done. ANOVA and Student's t-test were used for statistical analysis. The depressed cognitive function with chronic stress exposure and the increased anxiety-like behavior with both stress inductions were improved via both caffeine applications (pcaffeine pretreatments in chronic stressed rats, and chronic caffeine in acute stressed ones reduced the elevated myeloperoxidase activities (pcaffeine (pcaffeine (pcaffeine decreased SOD activity (pcaffeine. The increased anxiety-like behavior and depleted cognitive functions under stress conditions were improved with both acute and predominantly chronic caffeine pretreatments by decreasing oxidative damage parameters. Copyright © 2016 Elsevier Inc. All rights reserved.

  2. Anti-inflammatory activity of 3-thiazolyl coumarins

    International Nuclear Information System (INIS)

    Salar, U.; Khan, M.; Bakhtawar, A.; Kanwal, A.; Jabeen, A.; Faheem, A.

    2017-01-01

    3-Thiazolyl coumarins 1-33 along with coumarin scaffold (IC50 = 5.2 +- 0.2 mu g/mL) were evaluated for in vitro antiinflammatory activity. Activity of compounds was investigated by looking their influence on oxidative burst activity of zymosan stimulated whole blood phagocytes by using a luminol enhanced chemiluminescence technique. Ibuprofen was used as standard drug (IC50 = 54.2 +- 9.2 mu M). Four 3-thiazolyl coumarin derivatives 9 (IC50 = 31.0 +- 2.5 mu g/mL), 13 (IC50 = 27.1 +- 4.2 mu g/mL), 18 (IC50 = 5.6 +- 2.6 mu g/mL), and 29 (IC50 = 1.9 +- 1.0 mu g/mL) out of thirty-three demonstrated antiinflammatory activity as compared to the standard ibuprofen (IC50 = 11.2 +- 1.9 mu g/mL). Especially, compound 29 showed many folds better activity as compared to coumarin and standard ibuprofen. Structure-activity relationship was also established. It is worth-mentioning that active analogs 9, 13, 18, and 29 were found to be non-toxic on NIH-3T3 mouse fibroblast cell line. (author)

  3. An aptasensor for staphylococcus aureus based on nicking enzyme amplification reaction and rolling circle amplification.

    Science.gov (United States)

    Xu, Jingguo; Guo, Jia; Maina, Sarah Wanjiku; Yang, Yumeng; Hu, Yimin; Li, Xuanxuan; Qiu, Jiarong; Xin, Zhihong

    2018-05-15

    An ultra-sensitive aptamer-based biosensor for the detection of staphylococcus aureus was established by adopting the nicking enzyme amplification reaction (NEAR) and the rolling circle amplification (RCA) technologies. Aptamer-probe (AP), containing an aptamer and a probe sequence, was developed to act as the recognition unit of the biosensor, which was specifically bound to S. aureus. The probe was released from AP and initiated into the subsequent DNA amplification reactions where S. aureus was present, converting the detection of S. aureus to the investigation of probe oligonucleotide. The RCA amplification products contained a G-quadruplex motif and formed a three dimensional structure in presence of hemin. The G4/hemin complex showed horseradish peroxidase (HRP)-mimic activity and catalyzed the chemiluminescence reaction of luminol mediated by H 2 O 2 . The results showed that the established biosensor could detect S. aureus specifically with a good linear correlation at 5-10 4  CFU/mL. The signal values based on NEAR-RCA two-step cycle were boosted acutely, much higher than that relied on one-cycle magnification. The limit of detection (LoD) was determined to be as low as 5 CFU/mL. The established aptasensor exhibited a good discrimination of living against dead S. aureus, and can be applied to detect S. aureus in the food industry. Copyright © 2018 Elsevier Inc. All rights reserved.

  4. Adaptation of a load-inject valve for a flow injection chemiluminescence system enabling dual-reagent injection enhances understanding of environmental Fenton chemistry

    International Nuclear Information System (INIS)

    Jones, Matthew R.; Nightingale, Philp D.; Turner, Suzanne M.; Liss, Peter S.

    2013-01-01

    Graphical abstract: -- Highlights: •Measurement of multiple components of Fenton chemistry; Fe(II) and H 2 O 2 . •Rapid, quasi-simultaneous analysis enables calculation of environmental kinetics. •Low, nano to pico-molar detection limits with dual analyte analysis. •Able to measure complex matrix samples – organically enriched seawater. •Low cost system with appreciable sensitivity compared to single analyte analysis. -- Abstract: Environmental Fenton chemistry has been poorly constrained within the marine environment at a multi-component level. A simple, unique, reconfiguration of a flow-injection analytical system combined with luminol chemiluminescence allows quasi-simultaneously the measurement, using a single load-inject valve and a single photon multiplier tube, of reduced iron, Fe(II), and hydrogen peroxide. The system enables rapid, every 22 s, measurements with good accuracy at environmentally relevant concentrations, less than 5% relative standard deviations on both a 5 nM Fe(II) standard and a 60 nM hydrogen peroxide standard. Limits of detection were as low as 40 pM Fe(II) and 100 pM hydrogen peroxide. The system showed excellent capability by measuring from within an organic rich seawater the photochemically induced production of Fe(II) and hydrogen peroxide and their subsequent cycling and Fenton like interactions

  5. Decoupling peroxyacetyl nitrate from ozone in Chinese outflows observed at Gosan Climate Observatory

    Science.gov (United States)

    Han, Jihyun; Lee, Meehye; Shang, Xiaona; Lee, Gangwoong; Emmons, Louisa K.

    2017-09-01

    We measured peroxyacetyl nitrate (PAN) and other reactive species such as O3, NO2, CO, and SO2 with aerosols including mass, organic carbon (OC), and elemental carbon (EC) in PM2. 5 and K+ in PM1. 0 at Gosan Climate Observatory in Korea (33.17° N, 126.10° E) during 19 October-6 November 2010. PAN was determined through fast gas chromatography with luminol chemiluminescence detection at 425 nm every 2 min. The PAN mixing ratios ranged from 0.1 (detection limit) to 2.4 ppbv with a mean of 0.6 ppbv. For all measurements, PAN was unusually better correlated with PM2. 5 (Pearson correlation coefficient, γ = 0.79) than with O3 (γ = 0.67). In particular, the O3 level was highly elevated with SO2 at midnight, along with a typical midday peak when air was transported rapidly from the Beijing areas. The PAN enhancement was most noticeable during the occurrence of haze under stagnant conditions. In Chinese outflows slowly transported over the Yellow Sea, PAN gradually increased up to 2.4 ppbv at night, in excellent correlation with a concentration increase in PM2. 5 OC and EC, PM2. 5 mass, and PM1. 0 K+. The high K+ concentration and OC / EC ratio indicated that the air mass was impacted by biomass combustion. This study highlights PAN decoupling with O3 in Chinese outflows and suggests PAN as a useful indicator for diagnosing continental outflows and assessing their perturbation of regional air quality in northeast Asia.

  6. The oxidative burst reaction in mammalian cells depends on gravity.

    Science.gov (United States)

    Adrian, Astrid; Schoppmann, Kathrin; Sromicki, Juri; Brungs, Sonja; von der Wiesche, Melanie; Hock, Bertold; Kolanus, Waldemar; Hemmersbach, Ruth; Ullrich, Oliver

    2013-12-20

    Gravity has been a constant force throughout the Earth's evolutionary history. Thus, one of the fundamental biological questions is if and how complex cellular and molecular functions of life on Earth require gravity. In this study, we investigated the influence of gravity on the oxidative burst reaction in macrophages, one of the key elements in innate immune response and cellular signaling. An important step is the production of superoxide by the NADPH oxidase, which is rapidly converted to H2O2 by spontaneous and enzymatic dismutation. The phagozytosis-mediated oxidative burst under altered gravity conditions was studied in NR8383 rat alveolar macrophages by means of a luminol assay. Ground-based experiments in "functional weightlessness" were performed using a 2 D clinostat combined with a photomultiplier (PMT clinostat). The same technical set-up was used during the 13th DLR and 51st ESA parabolic flight campaign. Furthermore, hypergravity conditions were provided by using the Multi-Sample Incubation Centrifuge (MuSIC) and the Short Arm Human Centrifuge (SAHC). The results demonstrate that release of reactive oxygen species (ROS) during the oxidative burst reaction depends greatly on gravity conditions. ROS release is 1.) reduced in microgravity, 2.) enhanced in hypergravity and 3.) responds rapidly and reversible to altered gravity within seconds. We substantiated the effect of altered gravity on oxidative burst reaction in two independent experimental systems, parabolic flights and 2D clinostat / centrifuge experiments. Furthermore, the results obtained in simulated microgravity (2D clinorotation experiments) were proven by experiments in real microgravity as in both cases a pronounced reduction in ROS was observed. Our experiments indicate that gravity-sensitive steps are located both in the initial activation pathways and in the final oxidative burst reaction itself, which could be explained by the role of cytoskeletal dynamics in the assembly and function

  7. Escherichia coli LF82 differentially regulates ROS production and mucin expression in intestinal epithelial T84 cells: implication of NOX1.

    Science.gov (United States)

    Elatrech, Imen; Marzaioli, Viviana; Boukemara, Hanane; Bournier, Odile; Neut, Christel; Darfeuille-Michaud, Arlette; Luis, José; Dubuquoy, Laurent; El-Benna, Jamel; My-Chan Dang, Pham; Marie, Jean-Claude

    2015-05-01

    Increased reactive oxygen species (ROS) production is associated with inflamed ileal lesions in Crohn's disease colonized by pathogenic adherent-invasive Escherichia coli LF82. We investigated whether such ileal bacteria can modulate ROS production by epithelial cells, thus impacting on inflammation and mucin expression. Ileal bacteria from patients with Crohn's disease were incubated with cultured epithelial T84 cells, and ROS production was assayed using the luminol-amplified chemiluminescence method. The gentamicin protection assay was used for bacterial invasion of T84 cell. The expression of NADPH oxidase (NOX) subunits, mucin, and IL-8 was analyzed by quantitative real-time PCR and Western blots. Involvement of NOX and ROS was analyzed using diphenyleneiodonium (DPI) and N-acetylcysteine (NAC). Among different bacteria tested, only LF82 induced an increase of ROS production by T84 cells in a dose-dependent manner. This response was inhibited by DPI and NAC. Heat- or ethanol-attenuated LF82 bacteria and the mutant LF82ΔFimA, which does not express pili type 1 and poorly adheres to epithelial cells, did not induce the oxidative response. The LF82-induced oxidative response coincides with its invasion in T84 cells, and both processes were inhibited by DPI. Also, we observed an increased expression of NOX1 and NOXO1 in response to LF82 bacteria versus the mutant LF82ΔFimA. Furthermore, LF82 inhibited mucin gene expression (MUC2 and MUC5AC) in T84 cells while increasing the chemotactic IL-8 expression, both in a DPI-sensitive manner. Adherent-invasive E. coli LF82 induced ROS production by intestinal NADPH oxidase and altered mucin and IL-8 expression, leading to perpetuation of inflammatory lesions in Crohn's disease.

  8. Chemiluminescence immunoassay based on dual signal amplification strategy of Au/mesoporous silica and multienzyme functionalized mesoporous silica

    Energy Technology Data Exchange (ETDEWEB)

    Lin Jiehua, E-mail: linjiehua@qust.edu.cn [Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042 (China); Zhao Yue; Wei Zhijing; Wang Wei [Key Laboratory of Eco-chemical Engineering, Ministry of Education, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042 (China)

    2011-11-15

    Highlights: > The increased amount of monoclonal antibody in Au/SiO{sub 2} led to a wider linear range. > Due to the increased HRP tags in HRP-Ab{sub 2}/SiO{sub 2}, signal amplification achieved. > A simple dual amplification immunoassay achieved with flow injection analysis. - Abstract: A chemiluminescent dual signal amplification strategy for the determination of {alpha}-fetoprotein (AFP) was proposed based on a sandwich immunoassay format. Monoclonal antibody of AFP immobilized on the gold nanoparticles doped mesoporous SiO{sub 2} (Au/SiO{sub 2}) were prepared and used as a primary antibody. Horseradish peroxidase (HRP) and HRP-labeled secondary antibody (Ab{sub 2}) co-immobilized into the mesoporous SiO{sub 2} nanoparticles (HRP-Ab{sub 2}/SiO{sub 2}) were used as the labeled immunological probe. Due to the high ratio surface areas and pore volumes of the mesoporous SiO{sub 2}, not only the amount of AFP monoclonal antibody but also the amount of the modified HRP and Ab{sub 2} in HRP-Ab{sub 2}/SiO{sub 2} were largely increased. Thus the chemiluminescent signal was amplified by using the system of luminol and H{sub 2}O{sub 2} under the catalysis of HRP. Under the optimal conditions, two linear ranges for AFP were obtained from 0.01 to 0.5 ng mL{sup -1} and 0.5 to 100 ng mL{sup -1} with a detection limit of 0.005 ng mL{sup -1} (3{sigma}). The fabricated signal amplification strategy showed an excellent promise for sensitive detection of AFP and other tumor markers.

  9. Comparative antioxidant activity of cultivated and wild Vaccinium species investigated by EPR, human neutrophil burst and COMET assay.

    Science.gov (United States)

    Braga, P C; Antonacci, R; Wang, Y Y; Lattuada, N; Dal Sasso, M; Marabini, L; Fibiani, M; Lo Scalzo, R

    2013-01-01

    The Vaccinium (V.) spp. berries are considered a source of antioxidants, mainly belonging to polyphenols, specifically flavonoids and anthocyanins. Wild genotypes generally contain more antioxidants than cultivated counterparts. So, seven different antioxidants assays on extracts from cultivated and wild Vaccinium berries were performed, to evaluate their difference in terms of bioactivity on oxidative protection and minimum dosage to have a significant action. Four cell-free antioxidant assays (ABTS radical scavenging and electronic paramagnetic resonance using Fremy's salt, superoxide anion and hydroxyl radical), and three assays on human cells (two luminol amplified chemiluminescence, LACL, one on DNA damage, COMET) were used to measure the effects of cultivated blueberry (V. corymbosum) and wild bilberry (V. myrtillus) on the differently induced oxidative stress. Concentrations vs activity patterns were obtained by successive dilutions of extracts in order to identify both EC50 and minimum significant activity (MSA). All the assays (except for the hydroxyl radical scavenging) showed a good relationship mainly with anthocyanin and polyphenol content and the significant greater activity of wild Vaccinium extracts. In fact, LACL data gave an EC50 of 11.8 and an MSA of 5.2 g were calculated as fresh weight dosage in cultivated berries, compared with lower doses in wild berries, EC50 of 5.7 g and MSA of 3.4 g. Wild Vaccinium extracts averaged 3.04 and 2.40 fold more activity than cultivated extracts by EC50 and MSA, respectively. COMET assay confirmed the stronger action on DNA protection in wild samples.

  10. Synthesis and evaluation of the potential deleterious effects of ZnO nanomaterials (nanoneedles and nanoflowers) on blood components, including albumin, erythrocytes and human isolated primary neutrophils

    Energy Technology Data Exchange (ETDEWEB)

    Pastrello, Bruna [São Paulo State University (UNESP), Department of Chemistry, Faculty of Sciences (Brazil); Paracatu, Luana Chiquetto [São Paulo State University (UNESP), Department of Clinical Analysis, School of Pharmaceutical Sciences (Brazil); Carvalho Bertozo, Luiza de [São Paulo State University (UNESP), Department of Chemistry, Faculty of Sciences (Brazil); Paino, Iêda Maria Martinez [University of São Paulo (USP), Nanomedicine and Nanotoxicology Group, Physics Institute of São Carlos (IFSC) (Brazil); Lisboa-Filho, Paulo Noronha [São Paulo State University (UNESP), Department of Physics, Faculty of Sciences (Brazil); Ximenes, Valdecir Farias, E-mail: vfximenes@fc.unesp.br [São Paulo State University (UNESP), Department of Chemistry, Faculty of Sciences (Brazil)

    2016-07-15

    The application of zinc oxide (ZnO) nanoparticles in biomaterials has increased significantly in the recent years. Here, we aimed to study the potential deleterious effects of ZnO on blood components, including human serum albumin (HSA), erythrocytes and human isolated primary neutrophils. To test the influence of the morphology of the nanomaterials, ZnO nanoneedles (ZnO-nn) and nanoflowers (ZnO-nf) were synthesized. The zeta potential and mean size of ZnO-nf and ZnO-nn suspensions in phosphate-buffered saline were −10.73 mV and 3.81 nm and −5.27 mV and 18.26 nm, respectively. The incubation of ZnO with HSA did not cause its denaturation as verified by the absence of significant alterations in the intrinsic and extrinsic fluorescence and in the circular dichroism spectrum of the protein. The capacity of HSA as a drug carrier was not affected as verified by employing site I and II fluorescent markers. Neither type of ZnO was able to provoke the activation of neutrophils, as verified by lucigenin- and luminol-dependent chemiluminescence and by the extracellular release of hydrogen peroxide. ZnO-nf, but not ZnO-nn, induced the haemolysis of erythrocytes. In conclusion, our results reinforce the concept that ZnO nanomaterials are relatively safe for usage in biomaterials. A potential exception is the capacity of ZnO-nf to promote the lysis of erythrocytes, a discovery that shows the importance of the morphology in the toxicity of nanoparticles.

  11. fMLP-Induced IL-8 Release Is Dependent on NADPH Oxidase in Human Neutrophils

    Directory of Open Access Journals (Sweden)

    María A. Hidalgo

    2015-01-01

    Full Text Available N-Formyl-methionyl-leucyl-phenylalanine (fMLP and platelet-activating factor (PAF induce similar intracellular signalling profiles; but only fMLP induces interleukin-8 (IL-8 release and nicotinamide adenine dinucleotide phosphate reduced (NADPH oxidase activity in neutrophils. Because the role of ROS on IL-8 release in neutrophils is until now controversial, we assessed if NADPH oxidase is involved in the IL-8 secretions and PI3K/Akt, MAPK, and NF-κB pathways activity induced by fMLP. Neutrophils were obtained from healthy volunteers. IL-8 was measured by ELISA, IL-8 mRNA by qPCR, and ROS production by luminol-amplified chemiluminescence, reduction of ferricytochrome c, and FACS. Intracellular pH changes were detected by spectrofluorescence. ERK1/2, p38 MAPK, and Akt phosphorylation were analysed by immunoblotting and NF-κB was analysed by immunocytochemistry. Hydroxy-3-methoxyaceto-phenone (HMAP, diphenyleneiodonium (DPI, and siRNA Nox2 reduced the ROS and IL-8 release in neutrophils treated with fMLP. HMAP, DPI, and amiloride (a Na+/H+ exchanger inhibitor inhibited the Akt phosphorylation and did not affect the p38 MAPK and ERK1/2 activity. DPI and HMAP reduced NF-κB translocation induced by fMLP. We showed that IL-8 release induced by fMLP is dependent on NADPH oxidase, and ROS could play a redundant role in cell signalling, ultimately activating the PI3K/Akt and NF-κB pathways in neutrophils.

  12. A cascade amplification strategy based on rolling circle amplification and hydroxylamine amplified gold nanoparticles enables chemiluminescence detection of adenosine triphosphate.

    Science.gov (United States)

    Wang, Ping; Zhang, Tonghuan; Yang, Taoyi; Jin, Nan; Zhao, Yanjun; Fan, Aiping

    2014-08-07

    A highly sensitive and selective chemiluminescent (CL) biosensor for adenosine triphosphate (ATP) was developed by taking advantage of the ATP-dependent enzymatic reaction (ATP-DER), the powerful signal amplification capability of rolling circle amplification (RCA), and hydroxylamine-amplified gold nanoparticles (Au NPs). The strategy relies on the ability of ATP, a cofactor of T4 DNA ligase, to trigger the ligation-RCA reaction. In the presence of ATP, the T4 DNA ligase catalyzes the ligation reaction between the two ends of the padlock probe, producing a closed circular DNA template that initiates the RCA reaction with phi29 DNA polymerase and dNTP. Therein, many complementary copies of the circular template can be generated. The ATP-DER is eventually converted into a detectable CL signal after a series of processes, including gold probe hybridization, hydroxylamine amplification, and oxidative gold metal dissolution coupled with a simple and sensitive luminol CL reaction. The CL signal is directly proportional to the ATP level. The results showed that the detection limit of the assay is 100 pM of ATP, which compares favorably with those of other ATP detection techniques. In addition, by taking advantage of ATP-DER, the proposed CL sensing system exhibits extraordinary specificity towards ATP and could distinguish the target molecule ATP from its analogues. The proposed method provides a new and versatile platform for the design of novel DNA ligation reaction-based CL sensing systems for other cofactors. This novel ATP-DER based CL sensing system may find wide applications in clinical diagnosis as well as in environmental and biomedical fields.

  13. Titania nanotubes decorated with gold nanoparticles for electrochemiluminescent biosensing of glycosylated hemoglobin

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Qun [Institute of Analytical Chemistry, Dushu Lake Campus, Soochow University, Industrial Park, Suzhou, 215123 (China); College of Chemistry and Chemical Engineering, Nantong University, Nantong, 226019 (China); Tang, Shan [Institute of Analytical Chemistry, Dushu Lake Campus, Soochow University, Industrial Park, Suzhou, 215123 (China); Fang, Chen [Institute of Analytical Chemistry, Dushu Lake Campus, Soochow University, Industrial Park, Suzhou, 215123 (China); Second Affiliated Hospital, Soochow University, Suzhou, 215004 (China); Tu, Yi-Feng, E-mail: tuyf@suda.edu.cn [Institute of Analytical Chemistry, Dushu Lake Campus, Soochow University, Industrial Park, Suzhou, 215123 (China)

    2016-09-14

    A glycated hemoglobin (HbA1c) biosensor with high performance has been constructed in this work. Here the fructosyl amino acid oxidase was immobilized onto a pre-functionalized indium tin oxide glass with titania nanotubes decorated with gold nanoparticles. The property of nanocomposite was characterized by transmission electromicroscopy, scanning electron microscopy, electrochemistry and spectroscopy. Under the optimum conditions, fructosyl valine was detected by this biosensor. It exhibited a linear detection range from 4.0 × 10{sup −9} M to 7.2 × 10{sup −7} M, and a limit of detection for 3.8 × 10{sup −9} M at the signal-to-noise ratio of 3. Thus the HbA1c level in whole blood samples of healthy individuals or diabetic patients were evaluated with designed biosensor after pre-treatment of hydrolysis. The results of our detection were closely consistent with that of the standard method. At the same time, our biosensor has some advantages including high sensitivity, disposable usage and low cost, which implies its great promising application in point-of-care testing of HbA1c. - Highlights: • The enhanced electrochemiluminescence of luminol by AuNPs/TiNTs. • An ECL biosensor for HbA1c assay with ultra-high sensitivity. • A promising disposable device for diabetic diagnosis and treatment even for POCT. • The excellent regression of detected results with gold-standard method.

  14. Titania nanotubes decorated with gold nanoparticles for electrochemiluminescent biosensing of glycosylated hemoglobin

    International Nuclear Information System (INIS)

    Zhao, Qun; Tang, Shan; Fang, Chen; Tu, Yi-Feng

    2016-01-01

    A glycated hemoglobin (HbA1c) biosensor with high performance has been constructed in this work. Here the fructosyl amino acid oxidase was immobilized onto a pre-functionalized indium tin oxide glass with titania nanotubes decorated with gold nanoparticles. The property of nanocomposite was characterized by transmission electromicroscopy, scanning electron microscopy, electrochemistry and spectroscopy. Under the optimum conditions, fructosyl valine was detected by this biosensor. It exhibited a linear detection range from 4.0 × 10"−"9 M to 7.2 × 10"−"7 M, and a limit of detection for 3.8 × 10"−"9 M at the signal-to-noise ratio of 3. Thus the HbA1c level in whole blood samples of healthy individuals or diabetic patients were evaluated with designed biosensor after pre-treatment of hydrolysis. The results of our detection were closely consistent with that of the standard method. At the same time, our biosensor has some advantages including high sensitivity, disposable usage and low cost, which implies its great promising application in point-of-care testing of HbA1c. - Highlights: • The enhanced electrochemiluminescence of luminol by AuNPs/TiNTs. • An ECL biosensor for HbA1c assay with ultra-high sensitivity. • A promising disposable device for diabetic diagnosis and treatment even for POCT. • The excellent regression of detected results with gold-standard method.

  15. Multifunctional Dendrimer-templated Antibody Presentation on Biosensor Surfaces for Improved Biomarker Detection.

    Science.gov (United States)

    Han, Hye Jung; Kannan, Rangaramanujam M; Wang, Sunxi; Mao, Guangzhao; Kusanovic, Juan Pedro; Romero, Roberto

    2010-02-08

    Dendrimers, with their well-defined globular shape and a high density of functional groups, are ideal nanoscale materials for templating sensor surfaces. This work exploits dendrimers as a versatile platform for capturing biomarkers with improved sensitivity and specificity. Synthesis, characterization, fabrication, and functional validation of the dendrimer-based assay platform are described. Bifunctional hydroxyl/thiol functionalized G4-polyamidoamine (PAMAM) dendrimer is synthesized and immobilized on to the polyethylene-glycol (PEG)-functionalized assay plate by coupling PEG-maleimide and dendrimer thiol groups. Simultaneously, part of the dendrimer thiol groups are converted to hydrazide functionalities. The resulting dendrimer-modified surface is coupled to the capture antibody in the Fc region of the oxidized antibody. This preserves the orientation flexibility of the antigen binding region (Fv) of the antibody. To validate the approach, the fabricated plates are further used as a solid phase for developing a sandwich type ELISA to detect IL-6 and IL-1β, important biomarkers for early stages of chorioamnionitis. The dendrimer-modified plate provides assays with significantly enhanced sensitivity, lower nonspecific adsorption, and a detection limit of 0.13 pg ml -1 for IL-6 luminol detection and 1.15 pg ml -1 for IL-1β TMB detection, which are significantly better than those for the traditional ELISA. The assays were validated in human serum samples from normal (non-pregnant) woman and pregnant women with pyelonephritis. The specificity and the improved sensitivity of the dendrimer-based capture strategy could have significant implications for the detection of a wide range of cytokines and biomarkers since the capture strategy could be applied to multiplex microbead assays, conductometric immunosensors and field effect biosensors.

  16. Live Candida albicans Suppresses Production of Reactive Oxygen Species in Phagocytes▿ †

    Science.gov (United States)

    Wellington, Melanie; Dolan, Kristy; Krysan, Damian J.

    2009-01-01

    Production of reactive oxygen species (ROS) is an important aspect of phagocyte-mediated host responses. Since phagocytes play a crucial role in the host response to Candida albicans, we examined the ability of Candida to modulate phagocyte ROS production. ROS production was measured in the murine macrophage cell line J774 and in primary phagocytes using luminol-enhanced chemiluminescence. J774 cells, murine polymorphonuclear leukocytes (PMN), human monocytes, and human PMN treated with live C. albicans produced significantly less ROS than phagocytes treated with heat-killed C. albicans. Live C. albicans also suppressed ROS production in murine bone marrow-derived macrophages from C57BL/6 mice, but not from BALB/c mice. Live C. albicans also suppressed ROS in response to external stimuli. C. albicans and Candida glabrata suppressed ROS production by phagocytes, whereas Saccharomyces cerevisiae stimulated ROS production. The cell wall is the initial point of contact between Candida and phagocytes, but isolated cell walls from both heat-killed and live C. albicans stimulated ROS production. Heat-killed C. albicans has increased surface exposure of 1,3-β-glucan, a cell wall component that can stimulate phagocytes. To determine whether surface 1,3-β-glucan exposure accounted for the difference in ROS production, live C. albicans cells were treated with a sublethal dose of caspofungin to increase surface 1,3-β-glucan exposure. Caspofungin-treated C. albicans was fully able to suppress ROS production, indicating that suppression of ROS overrides stimulatory signals from 1,3-β-glucan. These studies indicate that live C. albicans actively suppresses ROS production in phagocytes in vitro, which may represent an important immune evasion mechanism. PMID:18981256

  17. D-lactic acid interferes with the effects of platelet activating factor on bovine neutrophils.

    Science.gov (United States)

    Alarcón, P; Conejeros, I; Carretta, M D; Concha, C; Jara, E; Tadich, N; Hidalgo, M A; Burgos, R A

    2011-11-15

    D-lactic acidosis occurs in ruminants, such as cattle, with acute ruminal acidosis caused by ingestion of excessive amounts of highly fermentable carbohydrates. Affected animals show clinical signs similar to those of septic shock, as well as acute laminitis and liver abscesses. It has been proposed that the inflammatory response and susceptibility to infection could both be caused by the inhibition of phagocytic mechanisms. To determine the effects of d-lactic acid on bovine neutrophil functions, we pretreated cells with different concentrations of D-lactic acid and measured intracellular pH using 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) and calcium flux using FLUO-3 AM-loaded neutrophils. Reactive oxygen species (ROS) production was measured using a luminol chemiluminescence assay, and MMP-9/gelatinase-B granule release was measured by zymography. CD11b and CD62L/l-selectin expression, changes in cell shape, superoxide anion production, phagocytosis of Escherichia coli-Texas red bioparticles, and apoptosis were all measured using flow cytometry. Our results demonstrated that D-lactic acid reduced ROS production, CD11b upregulation and MMP-9 release in bovine neutrophils treated with 100 nM platelet-activating factor (PAF). D-lactic acid induced MMP-9 release and, at higher concentrations, upregulated CD11b expression, decrease L-selectin expression, and induces late apoptosis. We concluded that D-lactic acid can interfere with neutrophil functions induced by PAF, leading to reduced innate immune responses during bacterial infections. Moreover, the increase of MMP-9 release and CD11b expression induced by 10mM D-lactic acid could promote an nonspecific neutrophil-dependent inflammatory reaction in cattle with acute ruminal acidosis. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Chemically Regulated ROS Generation from Gold Nanoparticles for Enzyme-Free Electrochemiluminescent Immunosensing.

    Science.gov (United States)

    Higashi, Yui; Mazumder, Joyotu; Yoshikawa, Hiroyuki; Saito, Masato; Tamiya, Eiichi

    2018-04-17

    In the present work, we report on an enzyme-free electrochemiluminescent (ECL) immunosensing scheme utilizing the catalytic generation of reactive oxygen species (ROS) from gold nanoparticles (AuNPs) (diameter ≥5 nm) dispersed in aqueous solutions of trishydroxymethylaminomethane (Tris). First, to examine this catalytic pathway in detail, the effects of various factors such as the AuNP size and concentration, dispersant type and concentration, and dissolved oxygen were investigated using the electrochemiluminescence (ECL) of luminol. It was found that the catalytic generation of ROS from AuNPs can be regulated chemically by altering conditions such as the type, concentration, and pH of the solution that the AuNPs are dispersed in. Under the best conditions studied in this work, the AuNPs displayed high catalytic activity toward ROS generation, with an estimated apparent turnover number per AuNP of 0.1 s -1 , comparable to those of several common peroxide-producing enzymes. Following these studies, this phenomenon was applied to develop a one-step enzyme-free ECL immunosensor based on sandwiching the target analyte using antibody-conjugated magnetic beads (MB) and AuNPs. Using IgA as a model analyte, the developed immunosensor was able to detect the target in the range of 1 ng/mL to 10 μg/mL, with the lower detection limit being comparable to those of commercial assays for the same target. Altering the antibodies used to modify the MB and AuNPs could further improve the detection limit as well as expand the applicability of this immunoassay to the detection of other analytes.

  19. Development and validation of the first assay method coupling liquid chromatography with chemiluminescence for the simultaneous determination of menadione and its thioether conjugates in rat plasma.

    Science.gov (United States)

    Elgawish, Mohamed Saleh; Shimomai, Chikako; Kishikawa, Naoya; Ohyama, Kaname; Wada, Mitsuhiro; Kuroda, Naotaka

    2013-09-16

    Menadione (2-methyl-1,4-naphthoquinone, MQ), a component of multivitamin drugs with antihemorrhagic, antineoplastic, and antimalarial activity, is frequently used to investigate quinone-induced cytotoxicity. The formation of MQ conjugates with glutathione (GSH) by Michael addition and subsequent biotransformation to yield N-acetyl-l-cysteine conjugates is believed to be an important detoxification process. However, the resulting conjugates, 2-methyl-3-(glutathione-S-yl)-1,4-naphthoquinone (MQ-GS) and 2-methyl-3-(N-acetyl-l-cysteine-S-yl)-1,4-naphthoquinone (MQ-NAC), retain the ability to redox cycle and to arylate cellular nucleophiles. Although the nephrotoxicity and hepatotoxicity of MQ-thiol conjugates have been reported in vitro, methods for their determination in vivo have yet to be published. Herein, a highly sensitive, simple, and selective HPLC-chemiluminescence (HPLC-CL) coupled method is reported, allowing for the first time the simultaneous determination of MQ, MQ-GS, and MQ-NAC in rat plasma after MQ administration. Our method exploits the unique redox characteristics of MQ, MQ-GS, and MQ-NAC to react with dithiothreitol (DTT) to liberate reactive oxygen species (ROS) which are detected by a CL assay using luminol as a CL probe. To verify the proposed mechanism, MQ-GS and MQ-NAC were synthetically prepared. Specimen preparation involved solid-phase extraction on an Oasis HLB cartridge followed by isocratic elution on an ODS column. No interference from endogenous substances was detected. Linearity was observed in the range of 5-120 nM for MQ-GS and MQ-NAC and 10-240 nM for MQ, with detection limits (S/N of 3) of 1.4, 0.8, and 128 fmol for MQ-GS, MQ-NAC, and MQ, respectively. The application of our method reported here is the first to extensively study the stability and reversibility of thiol-quinones.

  20. Evaluation of Sonochemiluminescence in a Phantom in the Presence of Protoporphyrin IX Conjugated to Nanoparticles

    Directory of Open Access Journals (Sweden)

    Ahmad Shanei

    2012-03-01

    Full Text Available Introduction When a liquid is irradiated with high-intensity and low-frequency ultrasound, acoustic cavitation occurs and there are some methods to determine and quantify this phenomenon. The existing methods for performing these experiments include sonochemiluminescence (SCL and chemical dosimetric methods. The particles in a liquid decrease the ultrasonic intensity threshold needed for cavitation onset. In this study, a new nanoconjugate made up of Protoporphyrin IX (PpIX and gold nanoparticles (GNP, i.e., Au-PpIX was used to provide nucleation sites for cavitation. The nonradiative relaxation time of PpIX in the presence of GNPs is longer than the similar time for PpIX without GNPs. This effect can be used in medical diagnostic and therapeutic applications. Materials and Methods The acoustic cavitation activity was investigated studying integrated SCL signal in the wavelength range of 400-500 nm in polyacrylamide gel phantom containing luminol using a cooled CCD spectrometer at different intensities of 1 MHz ultrasound. In order to confirm these results, a chemical dosimetric method was utilized, too. Results SCL signal level in gel phantom containing Au-PpIX was higher than the other phantoms. These results have been confirmed by the chemical dosimetric data. Conclusion This finding can be related to the existence of PpIX as a sensitizer and GNPs as cavitation nuclei. In other words, nanoparticles have acted as the sites for cavitation and have increased the cavitation rate. Another theory is that activation of PpIX has produced more free radicals and has enhanced the SCL signal level.

  1. Expression of IL-17A concentration and effector functions of peripheral blood neutrophils in food allergy hypersensitivity patients.

    Science.gov (United States)

    Żbikowska-Gotz, Magdalena; Pałgan, Krzysztof; Gawrońska-Ukleja, Ewa; Kuźmiński, Andrzej; Przybyszewski, Michał; Socha, Ewa; Bartuzi, Zbigniew

    2016-03-01

    Lymphocytes Th17 and other types of immune system cells produce IL17. By induction of cytokines and chemokines, the IL17 cytokine is involved in mechanisms of allergic reaction with participation of neutrophil granulocytes. It affects activation, recruitment, and migration of neutrophils to the tissues, regulating inflammatory reaction intensity. Excited neutrophils secrete inter alia elastase and reactive oxygen species (ROS)--significant mediators of inflammation process responsible for tissues damage.The aim of the study was to evaluate the concentrations of serum interleukin 17A, serum neutrophil elastase, and ROS production by neutrophils in patients with food allergy.The study included 30 patients with food allergy diagnosed based on interview, clinical symptoms, positive SPT, placebo controlled double-blind oral provocation trial, and the presence of asIgE in blood serum against selected food allergens using fluoro-immuno-enzymatic method FEIA UNICap 100. The control group consisted of 10 healthy volunteers. The concentrations of IL17A were determined in all patients using ELISA method with eBioscience kits, and elastase using BenderMed Systems kits. Chemiluminescence of non-stimulated neutrophils was evaluated using luminol-dependent kinetic method for 40 min on Luminoskan (Labsystems luminometer).The results of serum IL-17A concentrations and the values of chemiluminescence obtained by non-activated neutrophils, as well as elastase concentrations, were higher in patients with food allergic hypersensitivity compared to healthy volunteers.This study demonstrates a significance of IL-17A and activated neutrophil granulocytes in the course of diseases with food allergic hypersensitivity. © The Author(s) 2015.

  2. Fe(III)-TAML activator: a potent peroxidase mimic for chemiluminescent determination of hydrogen peroxide.

    Science.gov (United States)

    Vdovenko, Marina M; Demiyanova, Alexandra S; Kopylov, Kirill E; Sakharov, Ivan Yu

    2014-07-01

    Efforts to replace native peroxidase with its low molecular weight alternatives have stimulated a search for peroxidase mimetics. Herein we describe the oxidation of luminol with hydrogen peroxide catalyzed by commercially available Fe(III)-TAML activator 1a, which was shown to be a more active catalyst than hemin. At Fe(III)-TAML activator 1a use in chemiluminescent assay for H2O2 determination the detection limit value (3σ) of 5×10(-8)M was similar to the detection limit obtained with horseradish peroxidase (1×10(-7)M) and significantly lower than that obtained in the presence of hemin (6×10(-7)M). The linear ranges (R(2)=0.98) of the assay were 6×10(-8)-1×10(-6)M and 6×10(-7)-1×10(-6)M H2O2 for Fe(III)-TAML 1a and hemin, respectively. The CV values for Fe(III)-TAML 1a-based assay measured within the working range varied from 1.0% to 3.7% (n=4), whereas in the case of hemin -5.0% to 9.7% (n=4). Moreover, the sensitivity of Fe(III)-TAML 1a-based method was 56 and 5 times higher than that of hemin- and HRP-based methods, respectively. The obtained results open good perspectives to apply Fe(III)-TAML activator 1a in CL analytical methods instead of hemin, a traditionally used peroxidase mimetic. Copyright © 2014 Elsevier B.V. All rights reserved.

  3. Radiation-induced apoptosis of neural precursors cell cultures: early modulation of the response mediated by reactive oxygen and nitrogen species (ROS/RNS)

    Energy Technology Data Exchange (ETDEWEB)

    Gisone, P.; Dubner, D.; Robello, E.; Michelin, S.; Perez, M. R.

    2004-07-01

    Apoptosis, the typical mode of radiation-induced cell death in developing Central Nervous System (CNS), is closely related with the oxidative status. Enhanced radiation-induced generation of ROS/RNS has been observed after exposures to low radiation doses leading to cellular amplification of signal transduction and further molecular and cellular radiation-responses. Moreover Nitric oxide (NO) and hydroxyl radical are implicated in dopaminergic neurotoxicity in different parading. This study is an attempt to address the participation of radiation-induced free radicals production, the contribution of endogenous NO generation, and the excitonic pathway, in the radiation-induced apoptosis of neural cortical precursors. Cortical cells obtained from at 17 gestational day (gd) were irradiated with doses from 0,2 Gy to 2 Gy at a dose-rate of 0.3 Gy/m. A significant decrease of Luminol-dependent Chemiluminescence was evident 30 m after irradiation reaching basal levels at 120 m follow for a tendency to increasing values Incubations with Superoxide Dismatuse (SOD) decreased significantly the chemiluminescence in irradiated samples NO content estimated by measuring the stable products NO{sub 2} and NO{sub 3} released to the culture medium in the same period, has shown a time-dependent accumulation from 1 h post-irradiation. the apoptosis, determined 24 h post-irradiation by flow cytometry, morphology and DNA fragmentation revealed a dose-effect relationship with significant differences from 0.4 Gy. The samples pre-treated with 10 mM of N-acetyl cysteine (NAC) a precursor of intracellular GSH synthesis, shown a significant decrease of the apoptosis. Apoptosis was significantly increased in irradiated cells after inhibition of nitric oxide synthase (NOS) byL-NAME. We conclude that ROS/RNS play a pivotal role in the early signaling pathways leading to a radiation-induced cell death. (Author) 40 refs.

  4. Radiation-induced apoptosis of neural precursors cell cultures: early modulation of the response mediated by reactive oxygen and nitrogen species (ROS/RNS)

    International Nuclear Information System (INIS)

    Gisone, P.; Dubner, D.; Robello, E.; Michelin, S.; Perez, M. R.

    2004-01-01

    Apoptosis, the typical mode of radiation-induced cell death in developing Central Nervous System (CNS), is closely related with the oxidative status. Enhanced radiation-induced generation of ROS/RNS has been observed after exposures to low radiation doses leading to cellular amplification of signal transduction and further molecular and cellular radiation-responses. Moreover Nitric oxide (NO) and hydroxyl radical are implicated in dopaminergic neurotoxicity in different parading. This study is an attempt to address the participation of radiation-induced free radicals production, the contribution of endogenous NO generation, and the excitonic pathway, in the radiation-induced apoptosis of neural cortical precursors. Cortical cells obtained from at 17 gestational day (gd) were irradiated with doses from 0,2 Gy to 2 Gy at a dose-rate of 0.3 Gy/m. A significant decrease of Luminol-dependent Chemiluminescence was evident 30 m after irradiation reaching basal levels at 120 m follow for a tendency to increasing values Incubations with Superoxide Dismatuse (SOD) decreased significantly the chemiluminescence in irradiated samples NO content estimated by measuring the stable products NO 2 and NO 3 released to the culture medium in the same period, has shown a time-dependent accumulation from 1 h post-irradiation. the apoptosis, determined 24 h post-irradiation by flow cytometry, morphology and DNA fragmentation revealed a dose-effect relationship with significant differences from 0.4 Gy. The samples pre-treated with 10 mM of N-acetyl cysteine (NAC) a precursor of intracellular GSH synthesis, shown a significant decrease of the apoptosis. Apoptosis was significantly increased in irradiated cells after inhibition of nitric oxide synthase (NOS) byL-NAME. We conclude that ROS/RNS play a pivotal role in the early signaling pathways leading to a radiation-induced cell death. (Author) 40 refs

  5. Decoupling peroxyacetyl nitrate from ozone in Chinese outflows observed at Gosan Climate Observatory

    Directory of Open Access Journals (Sweden)

    J. Han

    2017-09-01

    Full Text Available We measured peroxyacetyl nitrate (PAN and other reactive species such as O3, NO2, CO, and SO2 with aerosols including mass, organic carbon (OC, and elemental carbon (EC in PM2. 5 and K+ in PM1. 0 at Gosan Climate Observatory in Korea (33.17° N, 126.10° E during 19 October–6 November 2010. PAN was determined through fast gas chromatography with luminol chemiluminescence detection at 425 nm every 2 min. The PAN mixing ratios ranged from 0.1 (detection limit to 2.4 ppbv with a mean of 0.6 ppbv. For all measurements, PAN was unusually better correlated with PM2. 5 (Pearson correlation coefficient, γ =  0.79 than with O3 (γ =  0.67. In particular, the O3 level was highly elevated with SO2 at midnight, along with a typical midday peak when air was transported rapidly from the Beijing areas. The PAN enhancement was most noticeable during the occurrence of haze under stagnant conditions. In Chinese outflows slowly transported over the Yellow Sea, PAN gradually increased up to 2.4 ppbv at night, in excellent correlation with a concentration increase in PM2. 5 OC and EC, PM2. 5 mass, and PM1. 0 K+. The high K+ concentration and OC ∕ EC ratio indicated that the air mass was impacted by biomass combustion. This study highlights PAN decoupling with O3 in Chinese outflows and suggests PAN as a useful indicator for diagnosing continental outflows and assessing their perturbation of regional air quality in northeast Asia.

  6. An "off-on" electrochemiluminescent biosensor based on DNAzyme-assisted target recycling and rolling circle amplifications for ultrasensitive detection of microRNA.

    Science.gov (United States)

    Zhang, Pu; Wu, Xiaoyan; Yuan, Ruo; Chai, Yaqin

    2015-03-17

    In this study, an off-on switching of a dual amplified electrochemiluminescence (ECL) biosensor based on Pb(2+)-induced DNAzyme-assisted target recycling and rolling circle amplification (RCA) was constructed for microRNA (miRNA) detection. First, the primer probe with assistant probe and miRNA formed Y junction which was cleaved with the addition of Pb(2+) to release miRNA. Subsequently, the released miRNA could initiate the next recycling process, leading to the generation of numerous intermediate DNA sequences (S2). Afterward, bare glassy carbon electrode (GCE) was immersed into HAuCl4 solution to electrodeposit a Au nanoparticle layer (depAu), followed by the assembly of a hairpin probe (HP). Then, dopamine (DA)-modified DNA sequence (S1) was employed to hybridize with HP, which switching off the sensing system. This is the first work that employs DA to quench luminol ECL signal, possessing the biosensor ultralow background signal. Afterward, S2 produced by the target recycling process was loaded onto the prepared electrode to displace S1 and served as an initiator for RCA. With rational design, numerous repeated DNA sequences coupling with hemin to form hemin/G-quadruplex were generated, which could exhibit strongly catalytic toward H2O2, thus amplified the ECL signal and switched the ON state of the sensing system. The liner range for miRNA detection was from 1.0 fM to 100 pM with a low detection limit down to 0.3 fM. Moreover, with the high sensitivity and specificity induced by the dual signal amplification, the proposed miRNA biosensor holds great potential for analysis of other interesting tumor markers.

  7. Variation of iron redox kinetics and its relation with molecular composition of standard humic substances at circumneutral pH.

    Science.gov (United States)

    Lee, Ying Ping; Fujii, Manabu; Kikuchi, Tetsuro; Terao, Koumei; Yoshimura, Chihiro

    2017-01-01

    Oxidation and reduction kinetics of iron (Fe) and proportion of steady-state Fe(II) concentration relative to total dissolved Fe (steady-state Fe(II) fraction) were investigated in the presence of various types of standard humic substances (HS) with particular emphasis on the photochemical and thermal reduction of Fe(III) and oxidation of Fe(II) by dissolved oxygen (O2) and hydrogen peroxide (H2O2) at circumneutral pH (pH 7-8). Rates of Fe(III) reduction were spectrophotometrically determined by a ferrozine method under the simulated sunlight and dark conditions, whereas rates of Fe(II) oxidation were examined in air-saturated solution using luminol chemiluminescence technique. The reduction and oxidation rate constants were determined to substantially vary depending on the type of HS. For example, the first-order rate constants varied by up to 10-fold for photochemical reduction and 7-fold for thermal reduction. The degree of variation in Fe(II) oxidation was larger for the H2O2-mediated reaction compared to the O2-mediated reaction (e.g., 15- and 3-fold changes for the former and latter reactions, respectively, at pH 8). The steady-state Fe(II) fraction under the simulated sunlight indicated that the Fe(II) fraction varies by up to 12-fold. The correlation analysis indicated that variation of Fe(II) oxidation is significantly associated with aliphatic content of HS, suggesting that Fe(II) complexation by aliphatic components accelerates Fe(II) oxidation. The reduction rate constant and steady-state Fe(II) fractions in the presence of sunlight had relatively strong positive relations with free radical content of HS, possibly due to the reductive property of radical semiquinone in HS. Overall, the findings in this study indicated that the Fe reduction and oxidation kinetics and resultant Fe(II) formation are substantially influenced by chemical properties of HS.

  8. A permeable cuticle is associated with the release of reactive oxygen species and induction of innate immunity.

    Directory of Open Access Journals (Sweden)

    Floriane L'Haridon

    2011-07-01

    Full Text Available Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS, including H(2O(2 and O(2 (-, are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI or catalase. H(2O(2 was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA. Accordingly, ABA biosynthesis mutants (aba2 and aba3 were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending

  9. The exploitation of inflammation in photodynamic therapy of pleural cancer (Conference Presentation)

    Science.gov (United States)

    Davis, Richard W.; Miller, Joann; Houser, Cassandra L.; Klampatsa, Astero; Jenkins, Tim; Cengel, Keith A.; Albelda, Steven M.; Busch, Theresa M.

    2017-02-01

    The onset of inflammation is a well-known physiology in tumors treated with photodynamic therapy (PDT). After PDT, the release of danger signals causes an influx of neutrophils, activation of dendritic cells, and an eventual initiation of the adaptive immune response. However, inflammation also lies at a crucial fulcrum for treatment outcome, as it can stimulate the expression of resistance factors. Therefore, effective treatment with PDT requires an understanding of the holistic contribution of inflammation. Within, we outline two means of studying tumor inflammation in the setting of PDT. Experiments are conducted in murine models of mesothelioma, including those that incorporate surgery prior to PDT or pleural propagation of the disease. First, we use a chemiluminescent agent, luminol, to detect the influx of neutrophils by in vivo molecular imaging. This longitudinal approach allows for the repeated non-invasive monitoring of PDT-induced neutrophil influx. Data clearly identify protocol-specific differences in tumor-associated neutrophil activity. Second, we describe the application of cone-beam CT to detect the fibrosis associated with murine orthotropic mesothelioma models. This approach incorporates novel methods in image segmentation to accurately identify diffuse disease in the thoracic cavity. These studies lay the foundation for future research to correlate long-term response with local PDT-induced inflammation. Such methods in monitoring of inflammation or tumor burden will enable characterization of the consequences of combinatorial therapy (e.g., intraoperative PDT). Resulting data will guide the selection of pharmacological agents or molecular imaging techniques that respectively exploit inflammation for therapeutic or monitoring purposes.

  10. Dual-readout Immunochromatographic Assay by Utilizing MnO2 Nanoflowers as the Unique Colorimetric/Chemiluminescent Probe

    Energy Technology Data Exchange (ETDEWEB)

    Ouyang, Hui; Lu, Quian; Wang, Wenwen; Song, Yang; Tu, Xinman; Zhu, Chengzhou; Smith, Jordan N.; Du, Dan; Fu, Zhifeng; Lin, Yuehe

    2018-04-17

    Manganese dioxide nanoflowers (MnO2 NFs) were synthesized and utilized as a dual readout probe to develop a novel immunochromatographic test strip (ITS) for detecting pesticide residues using chlorpyrifos as the model analyte. MnO2 NFs-labeled antibody for chlorpyrifos was employed as the signal tracer for conducting the ITS. After 10-min competitive immunoreaction, the tracer antibody was captured by the immobilized immunogen on test line in the test strip, resulting in the accumulation of MnO2 NFs. The accumulation of MnO2 NFs led to the appearance of brown color on the test line, which could be easily observed by the naked eye as a qualitative readout. Moreover, MnO2 NFs showed a remarkably enhancing effect on the luminol-H2O2 chemiluminescent (CL) system. Unlike peroxidase-like nanomaterials, the enhancing mechanism of MnO2 NFs was based on its oxidant activity to decompose H2O2 for forming reactive oxygen species. After initiating the CL system in the test zone, strong CL signal was collected as a quantitative readout to sensitively detect chlorpyrifos. Under optimal conditions, the linear range of chlorpyrifos was 0.1–50 ng/mL with a low detection limit of 0.033 ng/mL (S/N = 3). The reliability of the dual-readout ITS was successfully demonstrated by the application on traditional Chinese medicine and environmental water samples. Due to the simultaneous rapid-qualitative and sensitive-quantitative detection, the dual-readout protocol provides a promising strategy for rapid screening and field assay on various areas such as environmental monitoring, food safety and point-of-care testing.

  11. The oxidant-scavenging abilities in the oral cavity may be regulated by a collaboration among antioxidants in saliva, microorganisms, blood cells and polyphenols: a chemiluminescence-based study.

    Directory of Open Access Journals (Sweden)

    Isaac Ginsburg

    Full Text Available Saliva has become a central research issue in oral physiology and pathology. Over the evolution, the oral cavity has evolved the antioxidants uric acid, ascorbate reduced glutathione, plasma-derived albumin and antioxidants polyphenols from nutrients that are delivered to the oral cavity. However, blood cells extravasated from injured capillaries in gingival pathologies, or following tooth brushing and use of tooth picks, may attenuate the toxic activities of H2O2 generated by oral streptococci and by oxidants generated by activated phagocytes. Employing a highly sensitive luminol-dependent chemiluminescence, the DPPH radical and XTT assays to quantify oxidant-scavenging abilities (OSA, we show that saliva can strongly decompose both oxygen and nitrogen species. However, lipophilic antioxidant polyphenols in plants, which are poorly soluble in water and therefore not fully available as effective antioxidants, can nevertheless be solubilized either by small amounts of ethanol, whole saliva or also by salivary albumin and mucin. Plant-derived polyphenols can also act in collaboration with whole saliva, human red blood cells, platelets, and also with catalase-positive microorganisms to decompose reactive oxygen species (ROS. Furthermore, polyphenols from nutrient can avidly adhere to mucosal surfaces, are retained there for long periods and may function as a "slow-release devises" capable of affecting the redox status in the oral cavity. The OSA of saliva is due to the sum result of low molecular weight antioxidants, albumin, polyphenols from nutrients, blood elements and microbial antioxidants. Taken together, saliva and its antioxidants are considered regulators of the redox status in the oral cavity under physiological and pathological conditions.

  12. The Oxidant-Scavenging Abilities in the Oral Cavity May Be Regulated by a Collaboration among Antioxidants in Saliva, Microorganisms, Blood Cells and Polyphenols: A Chemiluminescence-Based Study

    Science.gov (United States)

    Ginsburg, Isaac; Kohen, Ron; Shalish, Miri; Varon, David; Shai, Ella; Koren, Erez

    2013-01-01

    Saliva has become a central research issue in oral physiology and pathology. Over the evolution, the oral cavity has evolved the antioxidants uric acid, ascorbate reduced glutathione, plasma-derived albumin and antioxidants polyphenols from nutrients that are delivered to the oral cavity. However, blood cells extravasated from injured capillaries in gingival pathologies, or following tooth brushing and use of tooth picks, may attenuate the toxic activities of H2O2 generated by oral streptococci and by oxidants generated by activated phagocytes. Employing a highly sensitive luminol-dependent chemiluminescence, the DPPH radical and XTT assays to quantify oxidant-scavenging abilities (OSA), we show that saliva can strongly decompose both oxygen and nitrogen species. However, lipophilic antioxidant polyphenols in plants, which are poorly soluble in water and therefore not fully available as effective antioxidants, can nevertheless be solubilized either by small amounts of ethanol, whole saliva or also by salivary albumin and mucin. Plant-derived polyphenols can also act in collaboration with whole saliva, human red blood cells, platelets, and also with catalase-positive microorganisms to decompose reactive oxygen species (ROS). Furthermore, polyphenols from nutrient can avidly adhere to mucosal surfaces, are retained there for long periods and may function as a “slow- release devises” capable of affecting the redox status in the oral cavity. The OSA of saliva is due to the sum result of low molecular weight antioxidants, albumin, polyphenols from nutrients, blood elements and microbial antioxidants. Taken together, saliva and its antioxidants are considered regulators of the redox status in the oral cavity under physiological and pathological conditions. PMID:23658797

  13. Glucose-induced effects and joker function of glucose: endocrine or genotoxic prevalence?

    Science.gov (United States)

    Berstein, L M; Vasilyev, D A; Poroshina, T E; Kovalenko, I G

    2006-10-01

    The steady increase in chronic "glycemic load" is characteristic for modern times. Among myriad of glucose functions, two principals can be emphasized: first, endocrine (in particular, ability to induce insulin secretion) and second, DNA-damaging related to formation of reactive oxygen species (ROS). It was suggested by us earlier that a shift in the ratio of mentioned functions reflects a possible "joker" role of glucose as an important modifier of human pathology. Therefore, we embarked on a study to investigate an individual effect of peroral glucose challenge on serum insulin level and ROS generation by mononuclears (luminol-dependent/latex-induced chemiluminescence) in 20 healthy people aged between 28-75. Concentrations of glucose, blood lipids, carbonylated proteins, malondialdehyde, leptin and TNF-alpha were determined as well. On the basis of received data two separate groups could be distinguished: one (n=8), in which glucose stimulation of ROS generation by mononuclears was increased and relatively prevailed over induction of insulin secretion (state of the so called glucose-induced genotoxicity, GIGT), and another (n=12), in which signs of GIGT were not revealed. People who belonged to the first group were characterized with a tendency to lower body mass index, blood leptin and cholesterol and to higher TNF-alpha concentration. Thus, if joker function of glucose is realized in "genotoxic mode", the phenotype (and probably genotype) of subjects may be rather distinctive to the one discovered in glucose-induced "endocrine prevalence". Whether such changes may serve as a pro-mutagenic or pro-endocrine basis for the rise of different chronic diseases or, rather, different features/aggressiveness of the same disease warrants further study.

  14. Effects of gadolinium oxide nanoparticles on the oxidative burst from human neutrophil granulocytes

    International Nuclear Information System (INIS)

    Abrikossova, Natalia; Skoglund, Caroline; Ahrén, Maria; Uvdal, Kajsa; Bengtsson, Torbjörn

    2012-01-01

    We have previously shown that gadolinium oxide (Gd 2 O 3 ) nanoparticles are promising candidates to be used as contrast agents in magnetic resonance (MR) imaging applications. In this study, these nanoparticles were investigated in a cellular system, as possible probes for visualization and targeting intended for bioimaging applications. We evaluated the impact of the presence of Gd 2 O 3 nanoparticles on the production of reactive oxygen species (ROS) from human neutrophils, by means of luminol-dependent chemiluminescence. Three sets of Gd 2 O 3 nanoparticles were studied, i.e. as synthesized, dialyzed and both PEG-functionalized and dialyzed Gd 2 O 3 nanoparticles. In addition, neutrophil morphology was evaluated by fluorescent staining of the actin cytoskeleton and fluorescence microscopy. We show that surface modification of these nanoparticles with polyethylene glycol (PEG) is essential in order to increase their biocompatibility. We observed that the as synthesized nanoparticles markedly decreased the ROS production from neutrophils challenged with prey (opsonized yeast particles) compared to controls without nanoparticles. After functionalization and dialysis, more moderate inhibitory effects were observed at a corresponding concentration of gadolinium. At lower gadolinium concentration the response was similar to that of the control cells. We suggest that the diethylene glycol (DEG) present in the as synthesized nanoparticle preparation is responsible for the inhibitory effects on the neutrophil oxidative burst. Indeed, in the present study we also show that even a low concentration of DEG, 0.3%, severely inhibits neutrophil function. In summary, the low cellular response upon PEG-functionalized Gd 2 O 3 nanoparticle exposure indicates that these nanoparticles are promising candidates for MR-imaging purposes. (paper)

  15. Internal Light Source-Driven Photoelectrochemical 3D-rGO/Cellulose Device Based on Cascade DNA Amplification Strategy Integrating Target Analog Chain and DNA Mimic Enzyme.

    Science.gov (United States)

    Lan, Feifei; Liang, Linlin; Zhang, Yan; Li, Li; Ren, Na; Yan, Mei; Ge, Shenguang; Yu, Jinghua

    2017-11-01

    In this work, a chemiluminescence-driven collapsible greeting card-like photoelectrochemical lab-on-paper device (GPECD) with hollow channel was demonstrated, in which target-triggering cascade DNA amplification strategy was ingeniously introduced. The GPECD had the functions of reagents storage and signal collection, and the change of configuration could control fluidic path, reaction time and alterations in electrical connectivity. In addition, three-dimentional reduced graphene oxide affixed Au flower was in situ grown on paper cellulose fiber for achieving excellent conductivity and biocompatibility. The cascade DNA amplification strategy referred to the cyclic formation of target analog chain and its trigger action to hybridization chain reaction (HCR), leading to the formation of numerous hemin/G-quadruplex DNA mimic enzyme with the presence of hemin. Subjected to the catalysis of hemin/G-quadruplex, the strong chemiluminiscence of luminol-H 2 O 2 system was obtained, which then was used as internal light source to excite photoactive materials realizing the simplification of instrument. In this analyzing process, thrombin served as proof-of-concept, and the concentration of target was converted into the DNA signal output by the specific recognition of aptamer-protein and target analog chain recycling. The target analog chain was produced in quantity with the presence of target, which further triggered abundant HCR and introduced hemin/G-quadruplex into the system. The photocurrent signal was obtained after the nitrogen-doped carbon dots sensitized ZnO was stimulated by chemiluminescence. The proposed GPECD exhibited excellent specificity and sensitivity toward thrombin with a detection limit of 16.7 fM. This judiciously engineered GPECD paved a luciferous way for detecting other protein with trace amounts in bioanalysis and clinical biomedicine.

  16. [Change of blood antioxidant capacity of experimental animals during nutritional correction under oxidative stress].

    Science.gov (United States)

    Basov, A A; Bykov, I M

    2013-01-01

    The effect of nutritional correction (a diet high in foods with antioxidant content) on blood parameters in laboratory animals with metabolic disorders associated with oxidative stress has been studied. In experimental models of laboratory animals (male rabbits weighing 3.5-4.0 kg, n = 40) with purulent septic diseases it has been demonstrated that the use of nutritive correction (replacement of 100 g of the cereal mixture through day on a mixture of cabbage 50 g, carrots 50 g, beet 25 g, apple 25 g, kiwi 10 g and garnet 10 g per 1 rabbit) is not inferior to its efficiency of glutathione use (2 g per day). The use of these antioxidants in laboratory animals significantly reduced the phenomenon of oxidative stress on the 5th day: blood antioxidant capacity significantly increased by 14.9 and 26.6%, and the area of the flash of luminol-dependent H2O2-induced chemiluminescence of blood plasma reduced by 44.2 and 48.6% in the experimental groups receiving respectively nutritive correction and glutathione. The low-molecula level of blood antioxidant capacity was restored and the balance of the activity of superoxide dismutase (decrease) and catalase (increase) was achieved on the 10th day of the experiment. These figures significantly (p < 0.05) differed from than in the group of animals receiving no antioxidant correction. The latter studied parameters of prooxidant-antioxidant system reached values comparable with those in intact animals (n = 10) only on the 30th day, confirming the advisability of appointing a complex antioxidant therapy.

  17. Monitoring of transient cavitation induced by ultrasound and intense pulsed light in presence of gold nanoparticles.

    Science.gov (United States)

    Sazgarnia, Ameneh; Shanei, Ahmad; Shanei, Mohammad Mahdi

    2014-01-01

    One of the most important challenges in medical treatment is invention of a minimally invasive approach in order to induce lethal damages to cancer cells. Application of high intensity focused ultrasound can be beneficial to achieve this goal via the cavitation process. Existence of the particles and vapor in a liquid decreases the ultrasonic intensity threshold required for cavitation onset. In this study, synergism of intense pulsed light (IPL) and gold nanoparticles (GNPs) has been investigated as a means of providing nucleation sites for acoustic cavitation. Several approaches have been reported with the aim of cavitation monitoring. We conducted the experiments on the basis of sonochemiluminescence (SCL) and chemical dosimetric methods. The acoustic cavitation activity was investigated by determining the integrated SCL signal acquired over polyacrylamide gel phantoms containing luminol in the presence and absence of GNPs in the wavelength range of 400-500 nm using a spectrometer equipped with cooled charged coupled devices (CCD) during irradiation by different intensities of 1 MHz ultrasound and IPL pulses. In order to confirm these results, the terephthalic acid chemical dosimeter was utilized as well. The SCL signal recorded in the gel phantoms containing GNPs at different intensities of ultrasound in the presence of intense pulsed light was higher than the gel phantoms without GNPs. These results have been confirmed by the obtained data from the chemical dosimetry method. Acoustic cavitation in the presence of GNPs and intense pulsed light has been suggested as a new approach designed for decreasing threshold intensity of acoustic cavitation and improving targeted therapeutic effects. Copyright © 2013 Elsevier B.V. All rights reserved.

  18. Decreased activity of neutrophils in the presence of diferuloylmethane (curcumin) involves protein kinase C inhibition.

    Science.gov (United States)

    Jancinová, Viera; Perecko, Tomás; Nosál, Radomír; Kostálová, Daniela; Bauerová, Katarína; Drábiková, Katarína

    2009-06-10

    Diferuloylmethane (curcumin) has been shown to act beneficially in arthritis, particularly through downregulated expression of proinflammatory cytokines and collagenase as well as through the modulated activities of T lymphocytes and macrophages. In this study its impact on activated neutrophils was investigated both in vitro and in experimental arthritis. Formation of reactive oxygen species in neutrophils was recorded on the basis of luminol- or isoluminol-enhanced chemiluminescence. Phosphorylation of neutrophil protein kinases C alpha and beta II was assessed by Western blotting, using phosphospecific antibodies. Adjuvant arthritis was induced in Lewis rats by heat-killed Mycobacterium butyricum. Diferuloylmethane or methotrexate was administered over a period of 28 days after arthritis induction. Under in vitro conditions, diferuloylmethane (1-100 microM) reduced dose-dependently oxidant formation both at extra- and intracellular level and it effectively reduced protein kinase C activation. Adjuvant arthritis was accompanied by an increased number of neutrophils in blood and by a more pronounced spontaneous as well as PMA (phorbol myristate acetate) stimulated chemiluminescence. Whereas the arthritis-related alterations in neutrophil count and in spontaneous chemiluminescence were not modified by diferuloylmethane, the increased reactivity of neutrophils to PMA was less evident in diferuloylmethane-treated animals. The effects of diferuloylmethane were comparable with those of methotrexate. Diferuloylmethane was found to be a potent inhibitor of neutrophil functions both in vitro and in experimental arthritis. As neutrophils are considered to be cells with the greatest capacity to inflict damage within diseased joints, the observed effects could represent a further mechanism involved in the antirheumatic activity of diferuloylmethane.

  19. Risk of ocular blood splatter during oculofacial plastic surgery.

    Science.gov (United States)

    Stacey, Andrew W; Czyz, Craig N; Kondapalli, Srinivas Sai A; Hill, Robert H; Everman, Kelly R; Cahill, Kenneth V; Foster, Jill A

    2015-01-01

    To assess intraoperative blood splatter to the ocular surface and adnexa during oculofacial surgery. Four surgeons and multiple assistants at three separate locations wore a total of 331 protective eye shields during 131 surgeries. Postoperatively, a luminol blood detection system was used to identify blood splatter on the shields. In the event of positive blood splatter, the total number of blood spots was counted. Controls were used to verify the blood detection protocol. A postoperative questionnaire was given to all surgeons and assistants after each case, and they were asked whether intraoperative blood splatter was noticed. Blood was detected on 61% of eye shields and in a total of 80% of surgical cases. However, only 2% of blood splatters were recognized intraoperatively by the surgical participants. There was no significant difference in the splatter rate between surgeons (64%), assistants (60%), and surgical technicians (58%) (p = 0.69). Shields worn during full-thickness eyelid procedures, direct brow lifting, orbitotomy with bony window, and orbital fracture repairs were more likely to be splattered (p = 0.03), and there was a significant difference between splatter rates among different surgeons (range, 29-90%; p = 0.0004), suggesting that blood splatter rate may be both procedure dependent and surgeon dependent. Mucocutaneous and transconjunctival transmission of human immunodeficiency virus and viral hepatitis has been documented. These results suggest that oculofacial plastic surgeons should consider eye protection for patients with known blood-borne diseases and in cases where blood splatter is expected. This precautionary practice is supported by the high incidence (98%) of undetected, intraoperative blood splatter.

  20. The Effects of Plantago major on the Activation of the Neutrophil Respiratory Burst

    Directory of Open Access Journals (Sweden)

    Elaine Reina

    2013-10-01

    Full Text Available Plantago major is a common plant that grows worldwide in temperate zones and is found in fields, lawns, and on the roadsides. Its leaves and seeds have been used in almost all parts of the world for centuries as a wound healer, analgesic, antioxidant, and antibiotic, as well as an immune system modulator, antiviral, antifungal, and anti-inflammatory agent. Baicalein and aucubin are the two most biologically active components of P. major, and both have been shown to have antioxidant, anti-inflammatory, and anticancer properties. Neutrophils have a pivotal role in wound healing and inflammation. Their principal mechanism of host defense is the killing of pathogens via the production of reactive oxygen species (ROS. The aim of the present study was to determine the in vitro effects of P. major extract, baicalein, and aucubin on human neutrophil respiratory burst activity. The cytotoxicity of the agents was assessed by lactate dehydrogenase (LDH assays. A standard luminol-dependent chemiluminescence (CL assay was utilized to monitor the respiratory burst of the neutrophils after exposure to P. major extract and its two active ingredients, baicalein and aucubin. Three replicates per group were included in each of the three runs of the experiments and analysis of variance (ANOVA was used for statistical analysis. P. major and baicalein were not toxic to the cells at any of the concentrations examined. Aucubin was toxic to the cells only at the highest concentration tested (P=0.0081. However, genistein was toxic to the cells at all of the concentrations examined except for the lowest concentration of 16.9 μg/ml (P=0.985. P. major (−0.10±0.11, aucubin (0.06±0.16, baicalein (−0.10±0.11, and genistein (−0.18±0.07 all significantly (P<0.0001 inhibited ROS production from the neutrophils. P. major extract inhibited neutrophil ROS production, as did aucubin and baicalein. Therefore, these components should be investigated further with relation to

  1. Radiosensitivity of the developing central nervous system: an important question in radiation protection

    International Nuclear Information System (INIS)

    Perez, Maria del R.; Gisone, Pablo A.; Dubner, Diana L.; Robello, Elizabeth; Michelin, Severino C.

    2004-01-01

    The high radiosensitivity of the human developing brain has been demonstrated by epidemiological data . The maximal incidence of microcephaly and mental retardation in the Hiroshima and Nagasaki cohort was observed among survivors exposed between 8 and 15 weeks of gestation. Experimental studies indicate that this period corresponds to gestational days (g.d.) 14-18 in the rat. Apoptosis, the typical mode of radiation-induced cell death in developing Central Nervous System (CNS), is closely related with the oxidative status. Enhanced radiation-induced generation of reactive oxygen and nitrogen species (ROS/RNS) has been observed after low-dose radiation exposure, leading to the amplification of signal transduction and further molecular and cellular radiation-responses. This study is an attempt to address the participation of ROS/RNS in the early radiation-induced apoptosis of neural cortical precursors. Cortical cells obtained from Wistar rat fetuses at 17 g.d. were irradiated with a dose of 2 Gy at a dose-rate of 0.3 Gy/minute. Nitric oxide (NO) content estimated by measuring the stable products NO 2 and NO 3 released to the culture medium, has shown a time-dependent accumulation from 30 min post-irradiation (pi). ROS/RNS content was determined by luminol-dependent chemiluminescence. A significant decrease of chemiluminescence was evident 30 min pi, reaching basal levels at 120 min, followed by an important augmentation by 4 hs after irradiation. The early decrease in ROS/RNS was not observed in samples irradiated in the presence of L-NAME, an inhibitor of nitric oxide synthase. Incubations with superoxide dismutase (SOD) and catalase (CAT) decreased significantly the chemiluminescence in irradiated samples. Apoptotic cell death, already evident from 4 hs pi., was significantly increased in irradiated cells after incubation with L-NAME. We conclude that ROS/RNS play a pivotal role in the early signaling pathway leading to a radiation-induced cell death of cortical

  2. Rate law of Fe(II) oxidation under low O2 conditions

    Science.gov (United States)

    Kanzaki, Yoshiki; Murakami, Takashi

    2013-12-01

    Despite intensive studies on Fe(II) oxidation kinetics, the oxidation rate law has not been established under low O2 conditions. The importance of Fe(II) oxidation under low O2 conditions has been recently recognized; for instance, the Fe(II)/Fe(III) compositions of paleosols, ancient soils formed by weathering, can produce a quantitative pattern of the atmospheric oxygen increase during the Paleoproterozoic. The effects of partial pressure of atmospheric oxygen (PO2) on the Fe(II) oxidation rate were investigated to establish the Fe(II) oxidation rate - PO2 relationships under low O2 conditions. All oxidation experiments were carried out in a glove box by introducing Ar gas at ∼10-5-∼10-4 atm of PO2, pH 7.57-8.09 and 22 °C. Luminol chemiluminescence was adopted to measure low Fe(II) concentrations (down to ∼2 nM). Combining previous data under higher PO2 conditions (10-3-0.2 atm) with the present data, the rate law for Fe(II) oxidation over a wide range of PO2 (10-5-0.2 atm) was found to be written as: d[Fe(II)]/dt=-k[Fe(II)][[]2 where the exponent of [O2], x, and the rate constant, k, change from x = 0.98 (±0.04) and log k = 15.46 (±0.06) at ∼6 × 10-3-0.2 atm of PO2 to x = 0.58 (±0.02) and log k = 13.41 (±0.03) at 10-5-∼6 × 10-3 atm of PO2. The most plausible mechanism that explains the change in x under low O2 conditions is that, instead of O2, oxygen-derived oxidants, H2O2 and to some extent, O2rad -, dominate the oxidation reactions at PO2. The rate law found in the present study requires us to reconsider distributions of Fe redox species at low PO2 in natural environments, especially in paleoweathering profiles, and may provide a deeper understanding of the evolution of atmospheric oxygen in the Precambrian.

  3. Resveratrol improves ifosfamide-induced Fanconi syndrome in rats

    International Nuclear Information System (INIS)

    Sehirli, Ozer; Sakarcan, Abdullah; Velioglu-Oguenc, Ayliz; Cetinel, Sule; Gedik, Nursal; Yegen, Berrak C.; Sener, Goeksel

    2007-01-01

    Regarding the mechanisms of ifosfamide (IFO)-induced urinary toxicity, several hypotheses have been put forward, among which oxidative stress and depletion of glutathione are suggested. This investigation elucidates the role of free radicals in IFO-induced toxicity and the protection by resveratrol, a natural phytoalexin. Wistar albino rats were injected intraperioneally with saline (0.9% NaCl; control), saline + resveratrol (RVT; 10 mg/kg/day), ifosfamide (IFO; 50 mg/kg/day) or IFO + RVT for 5 days. Urine was collected for 24 h during the 5th day, and at the 120th h after the first injections, animals were killed by decapitation and trunk blood was collected. Lactate dehydrogenase (LDH) activity, total antioxidant capacity (AOC) and pro-inflammatory cytokines TNF-α, IL-β and IL-6 were assayed in plasma samples. Kidney and bladder tissues were obtained for biochemical and histological analysis. Formation of reactive oxygen species in the tissue samples was monitored by using chemiluminescence (CL) technique with luminol and lucigenin probes. The results demonstrated that IFO induced a Fanconi syndrome characterized by increased urinary sodium, phosphate, glucose and protein, along with increased serum creatinine and urea levels. On the other hand, RVT markedly ameliorated the severity of renal dysfunction induced by IFO. Furthermore IFO caused a significant decrease in plasma AOC, which was accompanied with significant increases in the levels of the pro-inflammatory mediators and LDH activity, while RVT treatment reversed all these biochemical indices. In the saline-treated IFO group, glutathione levels were decreased significantly, while the malondialdehyde levels, myeloperoxidase activity and collagen content were increased in both tissues, which were in parallel with the increases in CL values. In the RVT-treated IFO group, all of these oxidant responses were prevented significantly. Our results suggest that IFO causes oxidative damage in the renal and bladder

  4. Variation of iron redox kinetics and its relation with molecular composition of standard humic substances at circumneutral pH.

    Directory of Open Access Journals (Sweden)

    Ying Ping Lee

    Full Text Available Oxidation and reduction kinetics of iron (Fe and proportion of steady-state Fe(II concentration relative to total dissolved Fe (steady-state Fe(II fraction were investigated in the presence of various types of standard humic substances (HS with particular emphasis on the photochemical and thermal reduction of Fe(III and oxidation of Fe(II by dissolved oxygen (O2 and hydrogen peroxide (H2O2 at circumneutral pH (pH 7-8. Rates of Fe(III reduction were spectrophotometrically determined by a ferrozine method under the simulated sunlight and dark conditions, whereas rates of Fe(II oxidation were examined in air-saturated solution using luminol chemiluminescence technique. The reduction and oxidation rate constants were determined to substantially vary depending on the type of HS. For example, the first-order rate constants varied by up to 10-fold for photochemical reduction and 7-fold for thermal reduction. The degree of variation in Fe(II oxidation was larger for the H2O2-mediated reaction compared to the O2-mediated reaction (e.g., 15- and 3-fold changes for the former and latter reactions, respectively, at pH 8. The steady-state Fe(II fraction under the simulated sunlight indicated that the Fe(II fraction varies by up to 12-fold. The correlation analysis indicated that variation of Fe(II oxidation is significantly associated with aliphatic content of HS, suggesting that Fe(II complexation by aliphatic components accelerates Fe(II oxidation. The reduction rate constant and steady-state Fe(II fractions in the presence of sunlight had relatively strong positive relations with free radical content of HS, possibly due to the reductive property of radical semiquinone in HS. Overall, the findings in this study indicated that the Fe reduction and oxidation kinetics and resultant Fe(II formation are substantially influenced by chemical properties of HS.

  5. Mitochondrial DNA maintenance is regulated in human hepatoma cells by glycogen synthase kinase 3β and p53 in response to tumor necrosis factor α.

    Science.gov (United States)

    Vadrot, Nathalie; Ghanem, Sarita; Braut, Françoise; Gavrilescu, Laura; Pilard, Nathalie; Mansouri, Abdellah; Moreau, Richard; Reyl-Desmars, Florence

    2012-01-01

    During chronic liver inflammation, up-regulated Tumor Necrosis Factor alpha (TNF-α) targets hepatocytes and induces abnormal reactive oxygen species (ROS) production responsible for mitochondrial DNA (mtDNA) alterations. The serine/threonine Glycogen Synthase Kinase 3 beta (GSK3β) plays a pivotal role during inflammation but its involvement in the maintenance of mtDNA remains unknown. The aim of this study was to investigate its involvement in TNF-α induced mtDNA depletion and its interrelationship with p53 a protein known to maintain mtDNA copy numbers. Using quantitative polymerase chain reaction (qPCR) we found that at 30 min in human hepatoma HepG2 cells TNF-α induced 0.55±0.10 mtDNA lesions per 10 Kb and a 52.4±2.8% decrease in mtDNA content dependent on TNF-R1 receptor and ROS production. Both lesions and depletion returned to baseline from 1 to 6 h after TNF-α exposure. Luminol-amplified chemiluminescence (LAC) was used to measure the rapid (10 min) and transient TNF-α induced increase in ROS production (168±15%). A transient 8-oxo-dG level of 1.4±0.3 ng/mg DNA and repair of abasic sites were also measured by ELISA assays. Translocation of p53 to mitochondria was observed by Western Blot and co-immunoprecipitations showed that TNF-α induced p53 binding to GSK3β and mitochondrial transcription factor A (TFAM). In addition, mitochondrial D-loop immunoprecipitation (mtDIP) revealed that TNF-α induced p53 binding to the regulatory D-loop region of mtDNA. The knockdown of p53 by siRNAs, inhibition by the phosphoSer(15)p53 antibody or transfection of human mutant active GSK3βS9A pcDNA3 plasmid inhibited recovery of mtDNA content while blockade of GSK3β activity by SB216763 inhibitor or knockdown by siRNAs suppressed mtDNA depletion. This study is the first to report the involvement of GSK3β in TNF-α induced mtDNA depletion. We suggest that p53 binding to GSK3β, TFAM and D-loop could induce recovery of mtDNA content through mtDNA repair.

  6. Mitochondrial DNA maintenance is regulated in human hepatoma cells by glycogen synthase kinase 3β and p53 in response to tumor necrosis factor α.

    Directory of Open Access Journals (Sweden)

    Nathalie Vadrot

    Full Text Available During chronic liver inflammation, up-regulated Tumor Necrosis Factor alpha (TNF-α targets hepatocytes and induces abnormal reactive oxygen species (ROS production responsible for mitochondrial DNA (mtDNA alterations. The serine/threonine Glycogen Synthase Kinase 3 beta (GSK3β plays a pivotal role during inflammation but its involvement in the maintenance of mtDNA remains unknown. The aim of this study was to investigate its involvement in TNF-α induced mtDNA depletion and its interrelationship with p53 a protein known to maintain mtDNA copy numbers. Using quantitative polymerase chain reaction (qPCR we found that at 30 min in human hepatoma HepG2 cells TNF-α induced 0.55±0.10 mtDNA lesions per 10 Kb and a 52.4±2.8% decrease in mtDNA content dependent on TNF-R1 receptor and ROS production. Both lesions and depletion returned to baseline from 1 to 6 h after TNF-α exposure. Luminol-amplified chemiluminescence (LAC was used to measure the rapid (10 min and transient TNF-α induced increase in ROS production (168±15%. A transient 8-oxo-dG level of 1.4±0.3 ng/mg DNA and repair of abasic sites were also measured by ELISA assays. Translocation of p53 to mitochondria was observed by Western Blot and co-immunoprecipitations showed that TNF-α induced p53 binding to GSK3β and mitochondrial transcription factor A (TFAM. In addition, mitochondrial D-loop immunoprecipitation (mtDIP revealed that TNF-α induced p53 binding to the regulatory D-loop region of mtDNA. The knockdown of p53 by siRNAs, inhibition by the phosphoSer(15p53 antibody or transfection of human mutant active GSK3βS9A pcDNA3 plasmid inhibited recovery of mtDNA content while blockade of GSK3β activity by SB216763 inhibitor or knockdown by siRNAs suppressed mtDNA depletion. This study is the first to report the involvement of GSK3β in TNF-α induced mtDNA depletion. We suggest that p53 binding to GSK3β, TFAM and D-loop could induce recovery of mtDNA content through mtDNA repair.

  7. Anticonvulsive and free radical scavenging actions of two herbs, Uncaria rhynchophylla (MIQ) Jack and Gastrodia elata Bl., in kainic acid-treated rats.

    Science.gov (United States)

    Hsieh, C L; Tang, N Y; Chiang, S Y; Hsieh, C T; Lin, J G

    1999-01-01

    Uncaria rhynchophylla (Miq.) Jack (UR) and Gastrodia elata BI. (GE) are traditional Chinese herbs that are usually used in combination to treat convulsive disorders, such as epilepsy, in China. The aim of this study was to compare the anticonvulsive and free radical scavenging activities of UR alone and UR in combination with GE in rats. For the in vitro studies, brain tissues from 6 male Sprague-Dawley (SD) rats were treated with 120 microg/ml kainic acid (KA), with or without varied concentrations of UR or UR plus GE. For the in vivo studies, male SD rats (6 per group) received intraperitoneal (i.p.) injection of KA 12 mg/kg to induce epileptic seizures and generation of free radicals, with or without oral administration of UR 1 g/kg alone or UR 1 g/kg plus GE 1 g/kg. Epileptic seizures were verified by behavioral observations, and electroencephalography (EEG) and electromyography (EMG) recordings. These results showed that UR alone decreased KA-induced lipid peroxide levels in vitro, whereas UR plus GE did not produce a greater effect than UR alone. UR significantly reduced counts of wet dog shakes (WDS), paw tremor (PT) and facial myoclonia (FM) in KA-treated rats and significantly delayed the onset time of WDS, from 27 min in the control group to 40 min in the UR group. UR plus GE did not inhibit seizures more effectively than UR alone, but did further prolong the onset time of WDS to 63 min (P < 0.05 vs. UR alone). UR alone reduced the levels of free radicals in vivo, as measured by lipid peroxidation in the brain and luminol-chemiluminescence (CL) counts and lucigenin-CL counts in the peripheral whole blood, but the combination of GE and UR did not reduce free radical levels more markedly than UR alone. In conclusion, our results indicate that UR has anticonvulsive and free radical scavenging activities, and UR combined with GE exhibit greater inhibition on the onset time of WDS than UR alone. These findings suggest that the anticonvulsive effects of UR and

  8. Diagnosis and therapy of dysfunctions of human leukocytes after irradiation. Final report; Diagnose und Therapie von Funktionsstoerungen menschlicher Leukozyten nach Bestrahlung. Abschlussbericht

    Energy Technology Data Exchange (ETDEWEB)

    Beuningen, D. van; Kaffenberger, W. [Sanitaetsakademie der Bundeswehr, Muenchen (Germany); Baaske, C.; Leipert, D.

    1999-08-01

    Experiments were performed with isolated human PMN and with the promyelocytic HL-60 cell line, induced to differentiate along the granulocytic lineage with dimethyl sulfoxid. The respiratory burst reaction was triggered with soluble stimuli (chemotactic agent: formylated tripeptide, f-MLP, immune complexes, or phorbol ester) and measured flow cytometrically or as chemiluminescence signals with indicator molecules dihydrorhodamine 123 or luminol, respectively. The presence of enzymes was postulated, verified with Western blotting, and their activities were inhibited pharmacologically, lipid 2{sup nd}-messengers (diacylglycerol) were measured by HPLC. Our results identify protein kinase (PK) C activity as the central element of signal transduction cascades involved in the activation of the NADPH oxidase. In addition, several phospholipases ({beta}, {gamma}), protein tyrosine- as well as protein serine/threonine kinases and DAG kinases in combination with phosphatidate phosphohydrolase contribute to intracellular signal transduction processes. For the first time, besides PKC activity new cytoplasmic/membrane-bound elements of signal transduction processes (DAG concentration and DAG kinase activity, phosphatidylinositol 3-kinase activity), and the FC{gamma} receptor-mediated respiratory burst of PMN were identified as radiosensitive ''targets'' for doses < 5 Gy. Theses results provide a basis for causally-oriented therapeutic approaches and could help to explain immunodeficiencies observed after exposure to ionizing radiation. (orig./MG) [German] Die intrazellulaeren Signaluebertragungsprozesse zur Aktivierung der NADPH-Oxidase sind Gegenstand der vorliegenden Untersuchung an bestrahlten PMN des Menschen und an einem PMN-Modell: HL-60 Promyelozyten, die mit Dimethylsulfoxid zur Differenzierung zu Neutrophilen-aehnlichen induziert wurden. Behandlung der Zellen mit loeslichen Rezeptor-abhaengigen oder -unabhaengigen Stimuli (Chemotaxin und

  9. Effect of β-1.3/1.6-D-glucan derived from oyster mushroom Pleurotus ostreatus on biometrical, haematological, biochemical, and immunological indices in rainbow trout (Oncorhynchus mykiss).

    Science.gov (United States)

    Dobsikova, Radka; Blahova, Jana; Franc, Ales; Jakubik, Juraj; Mikulikova, Ivana; Modra, Helena; Novotna, Kamila; Svobodova, Zdenka

    2012-01-01

    Effect of long-term oral administration of three different concentrations (0.5, 1.0, and 2.0%) of micronized β-1.3/1.6-D-glucan derived from oyster mushroom (Pleurotus ostreatus, Hiratake) on biometrical, haematological, biochemical, and immunological indices of half-year-old rainbow trout (Oncorhynchus mykiss) was assessed in the study. Rainbow trout were feed commercial feed pellets containing β-1.3/1.6-D-glucan in the concentrations of 0.5, 1.0, and 2.0% for 85 days. Biometrical indices consisted in total and standard length, body and liver weight, from which derived somatic parameters such as Fulton´s condition factor and hepatosomatic index were calculated. Haematological parameters were evaluated according to unified methods for haematological examination in fish. Plasma biochemical profile was analysed using biochemical analyser Konelab 20i and Easy Lyte Analyzer. A phagocyte cells metabolic activity (induced chemiluminescence of phagocytes) was determined as an immunological parameter by a microplate luminometric method on Immunotech LM-01T. No clinical signs of behavioral, respiratory, or neurologic distress were observed in rainbow trout. Fish showed normal feeding behavior. As for biometric parameters, no significant changes in total and standard length, body weight, liver weight, as well as in condition factor and hepatosomatic index of experimental and control fish were found. In the course of the study, weight gains in rainbow trout were similar and continuous. Shifts in PCV (pglucose, lactate, total protein, cholesterol, calcium, natrium, potassium (all p<0.05), albumins and chlorides (both p<0.01), as well as catalytic activities of ALT and AST (both p<0.05) were changed in the course of the study. A phagocyte cells metabolic activity (luminol-induced chemiluminescence) in rainbow trout was not altered by oyster mushroom β-1.3/1.6-D-glucan administration. After long-term oral administration of three concentrations of micronized β-1.3/1.6-D

  10. Potencial antioxidativo da lágrima de adultos jovens Tear antioxidant potential in young adults

    Directory of Open Access Journals (Sweden)

    Patrícia Ioschpe Gus

    2006-08-01

    Full Text Available OBJETIVO: Estudar os antioxidantes da lágrima humana, estimulada pelo corte de cebola e a possível influência dos hábitos de vida sobre estas medidas. MÉTODOS: A amostra consistiu de dez adultos jovens, que responderam questionário sobre o hábito de fumar, de ingerir bebidas alcoólicas, frutas, vegetais e cereais, de usar vitaminas e/ou drogas e de praticar exercícios. O potencial reativo antioxidante total (TRAP foi analisado por meio da quimioluminescência do luminol, a superóxido dismutase (SOD foi medida pela inibição do piragolol e a absorbância de H2O2 a 240 nm foi utilizada para identificar a catalase. RESULTADOS: A média ± DP dos valores de potencial reativo antioxidante total foi 33,8±11,5 µM e de superóxido dismutase foi 10,8±1,9 U/mL. Não foi identificada atividade da catalase. Detectou-se associação entre a prática regular de exercícios e aumento nos valores de potencial reativo antioxidante total (p=0,021, com diferença de 18,6 µM entre os indivíduos que se exercitavam pelo menos uma vez por semana e os sedentários. Sexo masculino e aumento na concentração de potencial reativo antioxidante total também se associaram estatisticamente (p=0,013, com diferença de 16,3 µM entre os sexos. Houve associação entre hábito tabágico e aumento na superóxido dismutase (p=0,041, com diferença de 3,3 U/mL entre fumantes de mais de cinco cigarros/dia e não fumantes. O uso de vitamina C também demonstrou associação com a superóxido dismutase (p=0,018; a diferença para os que tomavam vitamina C foi de 3,3 U/mL a mais. CONCLUSÃO: Os antioxidantes do lacrimejamento reflexo foram mensuráveis em adultos jovens, e diferentes variáveis parecem ter influenciado nos resultados.PURPOSE: To study the antioxidant status of human tears, stimulated by onion fumes and the possible influence of the life habits thereon were measured. METHODS: Subjects were ten healthy young adults, who answered a questionnaire about

  11. Estudio de la migración y de la actividad metabólica de los polimofonucleares neutrófilos en pacientes con periodontitis juvenil localizada Migration and metabolic activity of polymorphonuclear neutrophils in patients with localized juvenile periodontitis

    Directory of Open Access Journals (Sweden)

    Pablo Javier Patiño Grajales

    1993-03-01

    Full Text Available La periodontitis juvenil localizada (PJL se caracteriza por presentar una pérdida rápida de la inserción del tejido conectivo y el hueso alveolar alrededor de los primeros molares e incisivos de la dentición permanente. Esta Investigación se realizó para establecer una relación entre el comportamiento In vitro de los polimorfonucleares neutrófilos (PMN y los diferentes tipos clínicos de la enfermedad. Se estudiaron 21 pacientes con PJL clasificados en tres grupos: incipiente, moderada y avanzada. Antes de que los pacientes recibieran algún tipo de tratamiento odontológico, se les aislaron los neutrófilos y se estudió, In vitro, la quimiotaxis bajo agarosa y la actividad metabólica mediante la quimioluminiscencia dependiente de luminol (QLDL. Las mismas pruebas fueron practicadas en forma paralela a un grupo de individuos sanos como controles. Los pacientes con PJL avanzada tuvieron una disrninución significativa de la quimiotaxis (p=0.023. Su índice quimiotáctico fue de 9.4 :t 1.01 mientras el de los controles fue 11.9 :t 0.33. De otro lado, la QLDL de los PMN estimulados con forbol miristato acetato (PMA fue normal en todos los grupos, comparados con los controles; en contraste la estimulación de los PMN con zymosan opsonizado, evidenció una actividad metabólica significativamente disminuida en los pacientes con PJL incipiente y avanzada: el promedio de producción de energía lumínica en cpm para el grupo con PJL avanzada fue 703376:!: 74510 y para el de PJL incipiente fue 737000:!: 168610; en contraste en el grupo control fue 916597:!: 47418 (p=0.012. Estos hallazgos revelan que los pacientes con PJL conforman un grupo heterogéneo en cuanto a la función de los PMN. Para esclarecer la respuesta de los PMN en cada tipo de PJL, se hace necesario estudiar la vinculación entre los componentes genéticos, el medio ambiente y los fenotipos clínicos de la enfermedad.

    Anti-inflammatory and antioxidant effect of cerium dioxide nanoparticles immobilized on the surface of silica nanoparticles in rat experimental pneumonia.

    Science.gov (United States)

    Serebrovska, Z; Swanson, R J; Portnichenko, V; Shysh, A; Pavlovich, S; Tumanovska, L; Dorovskych, A; Lysenko, V; Tertykh, V; Bolbukh, Y; Dosenko, V

    2017-08-01

    A massage with the potent counter-inflammatory material, cerium dioxide nanoparticles, is promising and the antioxidant properties of CeO 2 are considered the main, if not the only, mechanism of this action. Nevertheless, the elimination of ceria nano-particles from the organism is very slow and there is a strong concern for toxic effect of ceria due to its accumulation. To overcome this problem, we engineered a combined material in which cerium nanoparticles were immobilized on the surface of silica nanoparticles (CeO 2 NP), which is shown to be easily removed from an organism and could be used as carriers for nano-ceria. In our study particle size was 220±5nm, Zeta-potential -4.5mV (in water), surface charge density -17.22μC/cm 2 (at pH 7). Thirty-six male Wistar rats, 5 months old and 250-290g were divided into four groups: 1) control; 2) CeO 2 NP treatment; 3) experimental pneumonia (i/p LPS injection, 1mg/kg); and 4) experimental pneumonia treated with CeO 2 NP (4 times during the study in dosage of 0.6mg/kg with an orogastric catheter). Gas exchange and pulmonary ventilation were measured four times: 0, 1, 3 and 24h after LPS injection in both untreated and CeO 2 NP-treated animals. The mRNA of TNF-α, Il-6, and CxCL2 were determined by RT-PCR. ROS-generation in blood plasma and lung tissue homogenates were measured by means of lucigenin- and luminol-enhanced chemiluminescence. Endotoxemia in the acute phase was associated with: (1) pathological changes in lung morphology; (2) increase of ROS generation; (3) enhanced expression of CxCL2; and (4) a gradual decrease of VO 2 and V E . CeO2 NP treatment of intact animals did not make any changes in all studied parameters except for a significant augmentation of VO 2 and V E. CeO 2 NP treatment of rats with pneumonia created positive changes in diminishing lung tissue injury, decreasing ROS generation in blood and lung tissue and decreasing pro-inflammatory cytokine expression (TNF-α, Il-6 and CxCL2). Oxygen