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Byunghee Park, Suk-Jung Choi
In the present study, a method was developed for detection of Vibrio parahaemolyticus based on a stationary liquid phase lab-on-a-chip (SLP LOC). The present SLP LOC comprises a sample chamber, washing chamber, and detection chamber connected by two channels. The method utilizes two types of particles: capture particles (CPs), which are magnetic nanoparticles functionalized with antibody; and labeling particles (LPs), which are silica nanoparticles functionalized with horseradish peroxidase and antibody. Samples were added to the sample chamber with CPs and LPs, forming a CP-bacteria-LP complex, and the complex was transported to the detection chamber containing chromogenic substrate solution...
December 1, 2016: Biosensors & Bioelectronics
Ya Li, Yanqing Li, Junli Zhao, Xiaojing Zheng, Qinwen Mao, Haibin Xia
Enzyme-linked immunosorbent assay (ELISA) has been one of the main methods for detecting an antigen in an aqueous sample for more than four decades. Nowadays, one of the biggest concerns for ELISA is still how to improve the sensitivity of the assay, and the luciferase-luciferin reaction system has been noticed as a new detection method with high sensitivity. In this study, a luciferin-luciferase reaction system was used as the detection method for a sandwich ELISA system. It was shown that this new system led to an increase in the detection sensitivity of at least two times when compared with the traditional horseradish peroxidase (HRP) detection method...
December 6, 2016: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
Masaki Yamaguchi, Shingo Sato
Horseradish peroxidase (HRP)-catalyzed oxidation of 2,2-bis[3-acethylfilicinic acid-5-yl]acetic acid (BAFA, 4) produces Dehydro-3,3'-diacetyl-5,5'-methylenedifilicinic acid (DDMF, 3). A new photometric hydrogen donor (4) for peroxidase (POD)-catalyzed oxidation was demonstrated to be potentially useful for spectrocolorimetric and spectrofluorometric determination of HRP. Our developed colorimetric (absorption at 483nm) and fluorometric (emission at 529nm) systems both gave a linear calibration curve for HRP (y=0...
November 13, 2016: Spectrochimica Acta. Part A, Molecular and Biomolecular Spectroscopy
Barbara S Janović, Milica Lj Mićić Vićovac, Zoran M Vujčić, Miroslava T Vujčić
Peroxidases (EC have enormous biotechnological applications. Usage of more abundant, basic isoforms of peroxidases in diagnostic kits and/or in immunochemistry has led to under exploitation and disregard of horseradish peroxidase (HRP) acidic isoforms. Therefore, acidic horseradish peroxidase (HRP-A) isoenzyme was used for the preparation of a biocatalyst with improved ability in dye decolorization. Ten biocatalysts were prepared by covalent binding of enzyme to chitosan and alginate, adsorption followed by cross-linking on inorganic support (aluminum oxide), and encapsulation in spherical calcium alginate beads via polyethylene glycol...
December 1, 2016: Environmental Science and Pollution Research International
Dongpo Xu, Guoquan Yan, Mingxia Gao, Chunhui Deng, Xiangmin Zhang
Glycosylation and phosphorylation as the commonest and most important posttranslational modifications of proteins play critical roles in biological processes. Specific and sensitive strategies have been developed to identify the glycosylation and phosphorylation of proteins by mass spectrometry but most of these methods have mainly focused on enriching glycopeptides or phosphopeptides separately, and few of them can enrich both of them. In this work, highly selective SiO2-NH2@TiO2 hollow microspheres were designed and synthesized for the simultaneous selective enrichment of both phosphopeptides and glycopeptides...
November 29, 2016: Analytical and Bioanalytical Chemistry
Feng-Bin Wang, Yi-Han Liao, Yao-Chen Wang
The inner thoracic cavity is lined by the parietal pleura, and the lung lobes are covered by the visceral pleura. The parietal and visceral plurae form the pleural cavity that has negative pressure within to enable normal respiration. The lung tissues are bilaterally innervated by vagal and spinal nerves, including sensory and motor components. This complicated innervation pattern has made it difficult to discern the vagal vs. spinal processes in the pulmonary visceral pleura. With and without vagotomy, we identified vagal nerve fibres and endings distributed extensively in the visceral pleura ('P'-type nerve endings) and triangular ligaments ('L'-type nerve endings) by injecting wheat germ agglutinin-horseradish peroxidase as a tracer into the nucleus of solitary tract or nodose ganglion of male Sprague-Dawley rats...
November 29, 2016: Journal of Anatomy
Yoshinori Mitsui
BACKGROUND: Since artesunate (ART) became a vital component of artemisinin (ARM)-based combination therapies for the treatment for malaria, counterfeit ART drugs have spread in regions of Southeast Asia and Africa. The consumption of counterfeit ART drugs has resulted in the death of many patients. Thus, evaluating the quality of ART drugs is needed. There are several methods for quantitating the ART content in tablets, the most common being a high-performance liquid chromatography. However, that method is hampered by the need for expensive equipment and a rather time-consuming process of extraction...
2016: Tropical Medicine and Health
Yue Lin, Mengmeng Zhao, Yajuan Guo, Xiaoming Ma, Fang Luo, Longhua Guo, Bin Qiu, Guonan Chen, Zhenyu Lin
In this work, 3,3',5,5'-tetramethylbenzidine(II) (TMB(2+)), derived from H2O2-horseradish peroxidase (HRP)-3,3',5,5'-tetramethylbenzidine (H2O2-HRP-TMB) reaction system, was used to etch AuNRs to generate different colors of solution. Many enzyme reactions are involved in the production of H2O2 (e.g., glucose can react with the dissolved oxygen in the presence of glucose oxidase (GOx) to produce H2O2). Given this information, a simple visual biosensor was developed in this study, with glucose as the example target...
November 25, 2016: Scientific Reports
J Reiling, K R Bridle, M Gijbels, F G Schaap, L Jaskowski, N Santrampurwala, L J Britton, C M Campbell, S W M Olde Damink, D H G Crawford, C H C Dejong, J Fawcett
INTRODUCTION: This study explored whether bacterial endotoxins, in the form of lipopolysaccharides (LPS), could have an injurious effect on the biliary tract in conjunction with ischemia. METHOD: Sixty-four rats were randomly assigned to four groups: sham operation (sham), 1mg/kg LPS i.p. (LPS), hepatic ischemia reperfusion (IR), and IR combined with LPS (IR+LPS). Following 1 or 6 hours of reperfusion, serum liver tests, bile duct histology, immunofluorescence microscopy (zonula occludens-1), bile composition (bile salts, phospholipids, lactate dehydrogenase), hepatic gene expression (bile salt transporters and inflammatory mediators) as well as serum and biliary cytokine concentrations were quantified and compared between the study groups...
November 23, 2016: Liver Transplantation
María Pedrero, F Javier Manuel de Villena, Cristina Muñoz-San Martín, Susana Campuzano, María Garranzo-Asensio, Rodrigo Barderas, José M Pingarrón
An amperometric magnetoimmunosensor for the determination of human p53 protein is described in this work using a sandwich configuration involving the covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic beads (HOOC-MBs) and incubation of the modified MBs with a mixture of the target protein and horseradish peroxidase-labeled antibody (HRP-anti-p53). The resulting modified MBs are captured by a magnet placed under the surface of a disposable carbon screen-printed electrode (SPCE) and the amperometric responses are measured at -0...
November 21, 2016: Biosensors
Yongmei Wu, Kai Sheng, Yongqin Liu, Qian Yu, Baoxian Ye
In this work, an ultrasensitive electrochemical microRNA detection strategy was developed based on porous palladium-modified horseradish peroxidase sphere (Pd@HRP) and target-induced assembly of DNAzyme. A highly loaded HRP sphere was prepared by covalent layer-by-layer assembly with CaCO3 as sacrificial template for the first time, and was further modified with porous Pd. Notably, Pd@HRP composite showed a good redox activity of HRP and electrocatalytic activity toward H2O2. The utilization of Pd@HRP as electrochemical signal indicator and enhancer to fabricate biosensor could avoid the need for additional redox mediator and amplify the detection sensitivity...
December 15, 2016: Analytica Chimica Acta
O Amor-Gutiérrez, E Costa Rama, A Costa-García, M T Fernández-Abedul
In this work we have developed an amperometric enzymatic biosensor in a paper-based platform with a mixed electrode configuration: carbon ink for the working electrode (WE) and metal wires (from a low-cost standard electronic connection) for reference (RE) and auxiliary electrodes (AE). A hydrophobic wax-defined paper area was impregnated with diluted carbon ink. Three gold-plated pins of the standard connection are employed, one for connecting the WE and the other two acting as RE and AE. The standard connection works as a clip in order to support the paper in between...
November 6, 2016: Biosensors & Bioelectronics
Balal Khalilzadeh, Nasrin Shadjou, Hadi Afsharan, Morteza Eskandani, Hojjatollah Nozad Charoudeh, Mohammad-Reza Rashidi
Introduction: Growing demands for ultrasensitive biosensing have led to the development of numerous signal amplification strategies. In this report, a novel electrochemiluminescence (ECL) method was developed for the detection and determination of caspase-3 activity based on reduced graphene oxide sheets decorated by gold nanoparticles as signal amplification element and horseradish peroxidase enzyme (HRP) as ECL intensity enhancing agent. Methods: The ECL intensity of the luminol was improved by using the streptavidin coated magnetic beads and HRP in the presence of hydrogen peroxide...
2016: BioImpacts: BI
Rui Chen, Xiaolin Huang, Juan Li, Shan Shan, Weihua Lai, Yonghua Xiong
Immunoassay is a powerful tool for rapid detection of food borne pathogens in food safety monitoring. However, conventional immunoassay always suffers from low sensitivity when it employs enzyme-catalyzing chromogenic substrates to generate colored molecules as signal outputs. In the present study, we report a novel fluorescence immunoassay for the sensitive detection of E. coli O157:H7 through combination of the ultrahigh bioactivity of catalase to hydrogen peroxide (H2O2) and H2O2-sensitive mercaptopropionic acid modified CdTe QDs (MPA-QDs) as a signal transduction...
December 1, 2016: Analytica Chimica Acta
Xiangheng Niu, Yanfang He, Jianming Pan, Xin Li, Fengxian Qiu, Yongsheng Yan, Libo Shi, Hongli Zhao, Minbo Lan
Nanosized materials acting as substitutes of natural enzymes are currently attracting significant research due to their stable enzyme-like characteristics, but some flaws of these nanozymes, including their limited catalytic rate and efficiency, need to be remedied to enable their wider applications. In this work, we verify for the first time the catalytic behavior of uncapped nanobranch-based CuS clews as a peroxidase mimic. XRD, XPS, SEM, and TEM proofs demonstrate that high-purity CuS clews composed of intertwined wires with abundant nanodendrites outside are successfully produced via a facile one-pot hydrothermal synthesis approach, with thiourea as both the sulfion source and the structure-directing agent...
December 1, 2016: Analytica Chimica Acta
Rui Yuan, Xiaolin Yu, Yuhong Zhang, Lulu Xu, Wei Cheng, Zhiguang Tu, Shijia Ding
Herein, a simple and novel fluorescence biosensing strategy has been developed for ultrasensitive determination of microRNA (miRNA) by combining target-triggered DNA nanoassembly on quantum dots (QDs) with DNAzyme-modulated double quenching of QDs. In presence of miRNA target, the target triggered catalytic hairpin assembly (CHA) amplification and powered highly efficient DNA nanoassembly on the surface of QDs, leading to exhibition of numerous G-quadruplexes close to the QDs. The G-quadruplex folded properly and bound hemin to form a stable G-quadruplex/hemin complex...
November 5, 2016: Biosensors & Bioelectronics
George Boyan, Erica Ehrhardt
The nervous system of the antenna of the grasshopper Schistocerca gregaria consists of two nerve tracts in which sensory cells project their axons to the brain. Each tract is pioneered early in embryogenesis by a pair of identified cells located apically in the antennal lumen. The pioneers are thought to originate in the epithelium of the antenna and then delaminate into the lumen where they commence axogenesis. However, unambiguous molecular identification of these cells in the epithelium, of an identifiable precursor, and of their mode of generation has been lacking...
November 10, 2016: Development Genes and Evolution
Junlin Wen, Junhua Chen, Li Zhuang, Shungui Zhou
An ultrasensitive and high-throughput nucleic acid detection system, termed as strand displacement reaction-enzyme linked immunosorbent assay (SDR-ELISA), has been developed on the basis of antibody-like DNA nanostructures. Three digoxigenin or biotin modified hairpin probes are utilized to construct antibody-like DNA nanostructures that feature affinity toward streptavidin and anti-digoxigenin antibody via isothermal target-triggered SDR amplification. These antibody-like nanostructures have been employed to conjugate horseradish-peroxidase-labeled anti-digoxigenin antibody with streptavidin that is immobilized on microliter plate wells for enzyme-linked colorimetric assay...
November 2, 2016: Biosensors & Bioelectronics
Xiaoqiang Liu, Pepipei Liu, Xiaohe Huo, Xiuhua Liu, Jin Liu
TiO2 nanosheets (TNSs) were synthesized and deposited on multi-wall carbon nanotubes (MWCNTs) to form a nano-composite through a hydrothermal method, followed by the characterization with various spectroscopic and microscopic techniques. The TNS-MWCNT composite was then applied as not only an electrode scaffold to immobilize primary antibody, but also as a carrier to load secondary antibody and horseradish peroxidase (HRP). In both cases, bis(sulfosuccinimidyl) suberate sodium salt acted as an amino cross-linker to covalently bind the biomolecules on TNS-MWCNT composite through their surface primary amino groups...
November 23, 2016: Analytica Chimica Acta
Shunxiang Gao, Xin Zheng, Bo Hu, Mingjuan Sun, Jihong Wu, Binghua Jiao, Lianghua Wang
In this study, we coupled biolayer interferometry (BLI) with competitive binding assay through an enzyme-linked aptamer and developed a real-time, ultra-sensitive, rapid quantitative method for detection of the marine biotoxin palytoxin. Horseradish peroxidase-labeled aptamers were used as biorecognition receptors to competitively bind with palytoxin, which was immobilized on the biosensor surface. The palytoxin: horseradish peroxidase-aptamer complex was then submerged in a 3,3'-diaminobenzidine solution, which resulted in formation of a precipitated polymeric product directly on the biosensor surface and a large change in the optical thickness of the biosensor layer...
September 25, 2016: Biosensors & Bioelectronics
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