A Novel Arc Fault Detector for Early Detection of Electrical Fires.

Science.gov (United States)

Yang, Kai; Zhang, Rencheng; Yang, Jianhong; Liu, Canhua; Chen, Shouhong; Zhang, Fujiang

2016-04-09

Arc faults can produce very high temperatures and can easily ignite combustible materials; thus, they represent one of the most important causes of electrical fires. The application of arc fault detection, as an emerging early fire detection technology, is required by the National Electrical Code to reduce the occurrence of electrical fires. However, the concealment, randomness and diversity of arc faults make them difficult to detect. To improve the accuracy of arc fault detection, a novel arc fault detector (AFD) is developed in this study. First, an experimental arc fault platform is built to study electrical fires. A high-frequency transducer and a current transducer are used to measure typical load signals of arc faults and normal states. After the common features of these signals are studied, high-frequency energy and current variations are extracted as an input eigenvector for use by an arc fault detection algorithm. Then, the detection algorithm based on a weighted least squares support vector machine is designed and successfully applied in a microprocessor. Finally, an AFD is developed. The test results show that the AFD can detect arc faults in a timely manner and interrupt the circuit power supply before electrical fires can occur. The AFD is not influenced by cross talk or transient processes, and the detection accuracy is very high. Hence, the AFD can be installed in low-voltage circuits to monitor circuit states in real-time to facilitate the early detection of electrical fires.

A Novel Arc Fault Detector for Early Detection of Electrical Fires

Science.gov (United States)

Yang, Kai; Zhang, Rencheng; Yang, Jianhong; Liu, Canhua; Chen, Shouhong; Zhang, Fujiang

2016-01-01

Arc faults can produce very high temperatures and can easily ignite combustible materials; thus, they represent one of the most important causes of electrical fires. The application of arc fault detection, as an emerging early fire detection technology, is required by the National Electrical Code to reduce the occurrence of electrical fires. However, the concealment, randomness and diversity of arc faults make them difficult to detect. To improve the accuracy of arc fault detection, a novel arc fault detector (AFD) is developed in this study. First, an experimental arc fault platform is built to study electrical fires. A high-frequency transducer and a current transducer are used to measure typical load signals of arc faults and normal states. After the common features of these signals are studied, high-frequency energy and current variations are extracted as an input eigenvector for use by an arc fault detection algorithm. Then, the detection algorithm based on a weighted least squares support vector machine is designed and successfully applied in a microprocessor. Finally, an AFD is developed. The test results show that the AFD can detect arc faults in a timely manner and interrupt the circuit power supply before electrical fires can occur. The AFD is not influenced by cross talk or transient processes, and the detection accuracy is very high. Hence, the AFD can be installed in low-voltage circuits to monitor circuit states in real-time to facilitate the early detection of electrical fires. PMID:27070618

A New Anomaly Detection System for School Electricity Consumption Data

Directory of Open Access Journals (Sweden)

Wenqiang Cui

2017-11-01

Full Text Available Anomaly detection has been widely used in a variety of research and application domains, such as network intrusion detection, insurance/credit card fraud detection, health-care informatics, industrial damage detection, image processing and novel topic detection in text mining. In this paper, we focus on remote facilities management that identifies anomalous events in buildings by detecting anomalies in building electricity consumption data. We investigated five models within electricity consumption data from different schools to detect anomalies in the data. Furthermore, we proposed a hybrid model that combines polynomial regression and Gaussian distribution, which detects anomalies in the data with 0 false negative and an average precision higher than 91%. Based on the proposed model, we developed a data detection and visualization system for a facilities management company to detect and visualize anomalies in school electricity consumption data. The system is tested and evaluated by facilities managers. According to the evaluation, our system has improved the efficiency of facilities managers to identify anomalies in the data.

A universal DNA-based protein detection system.

Science.gov (United States)

Tran, Thua N N; Cui, Jinhui; Hartman, Mark R; Peng, Songming; Funabashi, Hisakage; Duan, Faping; Yang, Dayong; March, John C; Lis, John T; Cui, Haixin; Luo, Dan

2013-09-25

Protein immune detection requires secondary antibodies which must be carefully selected in order to avoid interspecies cross-reactivity, and is therefore restricted by the limited availability of primary/secondary antibody pairs. Here we present a versatile DNA-based protein detection system using a universal adapter to interface between IgG antibodies and DNA-modified reporter molecules. As a demonstration of this capability, we successfully used DNA nano-barcodes, quantum dots, and horseradish peroxidase enzyme to detect multiple proteins using our DNA-based labeling system. Our system not only eliminates secondary antibodies but also serves as a novel method platform for protein detection with modularity, high capacity, and multiplexed capability.

[Detection of protein-protein interactions by FRET and BRET methods].

Science.gov (United States)

Matoulková, E; Vojtěšek, B

2014-01-01

Nowadays, in vivo protein-protein interaction studies have become preferable detecting meth-ods that enable to show or specify (already known) protein interactions and discover their inhibitors. They also facilitate detection of protein conformational changes and discovery or specification of signaling pathways in living cells. One group of in vivo methods enabling these findings is based on fluorescent resonance energy transfer (FRET) and its bio-luminescent modification (BRET). They are based on visualization of protein-protein interactions via light or enzymatic excitation of fluorescent or bio-luminescent proteins. These methods allow not only protein localization within the cell or its organelles (or small animals) but they also allow us to quantify fluorescent signals and to discover weak or strong interaction partners. In this review, we explain the principles of FRET and BRET, their applications in the characterization of protein-protein interactions and we describe several findings using these two methods that clarify molecular and cellular mechanisms and signals related to cancer biology.

Effect of External Electric Field Stress on Gliadin Protein Conformation

OpenAIRE

Singh, Ashutosh; Munshi, Shirin; Raghavan, Vijaya

2013-01-01

A molecular dynamic (MD) modeling approach was applied to evaluate the effect of external electric field on gliadin protein structure and surface properties. Static electric field strengths of 0.001 V/nm and 0.002 V/nm induced conformational changes in the protein but had no significant effect on its surface properties. The study of hydrogen bond evolution during the course of simulation revealed that the root mean square deviation, radius of gyration and secondary structure formation, all de...

A dielectrophoresis-impedance method for protein detection and analysis

Directory of Open Access Journals (Sweden)

Ahmad Sabry Mohamad

2017-01-01

Full Text Available Dielectrophoresis (DEP has increasingly been used for the assessment of the electrical properties of molecular scale objects including proteins, DNA, nanotubes and nanowires. However, whilst techniques have been developed for the electrical characterisation of frequency-dependent DEP response, biomolecular study is usually limited to observation using fluorescent markers, limiting its applicability as a characterisation tool. In this paper we present a label-free, impedance-based method of characterisation applied to the determination of the electrical properties of colloidal protein molecules, specifically Bovine Serum Albumin (BSA. By monitoring the impedance between electrodes as proteins collect, it is shown to be possible to observe multi-dispersion behaviour. A DEP dispersion exhibited at 400 kHz is attributable to the orientational dispersion of the molecule, whilst a second, higher-frequency dispersion is attributed to a Maxwell-Wagner type dispersion; changes in behaviour with medium conductivity suggest that this is strongly influenced by the electrical double layer surrounding the molecule.

A dielectrophoresis-impedance method for protein detection and analysis

Science.gov (United States)

Mohamad, Ahmad Sabry; Hamzah, Roszymah; Hoettges, Kai F.; Hughes, Michael Pycraft

2017-01-01

Dielectrophoresis (DEP) has increasingly been used for the assessment of the electrical properties of molecular scale objects including proteins, DNA, nanotubes and nanowires. However, whilst techniques have been developed for the electrical characterisation of frequency-dependent DEP response, biomolecular study is usually limited to observation using fluorescent markers, limiting its applicability as a characterisation tool. In this paper we present a label-free, impedance-based method of characterisation applied to the determination of the electrical properties of colloidal protein molecules, specifically Bovine Serum Albumin (BSA). By monitoring the impedance between electrodes as proteins collect, it is shown to be possible to observe multi-dispersion behaviour. A DEP dispersion exhibited at 400 kHz is attributable to the orientational dispersion of the molecule, whilst a second, higher-frequency dispersion is attributed to a Maxwell-Wagner type dispersion; changes in behaviour with medium conductivity suggest that this is strongly influenced by the electrical double layer surrounding the molecule.

Electric-field-induced superconductivity detected by magnetization measurements of an electric-double-layer capacitor

International Nuclear Information System (INIS)

Kasahara, Yuichi; Takeuchi, Yuki; Ye, Jianting; Yuan, Hongtao; Shimotani, Hidekazu; Iwasa, Yoshihiro; Nishimura, Takahiro; Sato, Tatsuya

2010-01-01

We report evidence for superconductivity induced by the application of strong electric fields onto the surface of a band insulator, ZrNCl, provided by the observation of a shielding diamagnetic signal. We introduced an electric-double-layer capacitor configuration and in situ magnetization measurements at low temperatures as a method to detect the novel electric-field-induced superconducting state. The results showed excellent agreement with a previous report using a transistor configuration, demonstrating that the present technique is a novel method for investigating the nonequilibrium phase induced by electric fields. (author)

In-channel electrochemical detection in the middle of microchannel under high electric field.

Science.gov (United States)

Kang, Chung Mu; Joo, Segyeong; Bae, Je Hyun; Kim, Yang-Rae; Kim, Yongseong; Chung, Taek Dong

2012-01-17

We propose a new method for performing in-channel electrochemical detection under a high electric field using a polyelectrolytic gel salt bridge (PGSB) integrated in the middle of the electrophoretic separation channel. The finely tuned placement of a gold working electrode and the PGSB on an equipotential surface in the microchannel provided highly sensitive electrochemical detection without any deterioration in the separation efficiency or interference of the applied electric field. To assess the working principle, the open circuit potentials between gold working electrodes and the reference electrode at varying distances were measured in the microchannel under electrophoretic fields using an electrically isolated potentiostat. In addition, "in-channel" cyclic voltammetry confirmed the feasibility of electrochemical detection under various strengths of electric fields (∼400 V/cm). Effective separation on a microchip equipped with a PGSB under high electric fields was demonstrated for the electrochemical detection of biological compounds such as dopamine and catechol. The proposed "in-channel" electrochemical detection under a high electric field enables wider electrochemical detection applications in microchip electrophoresis.

Aggregation of egg white proteins with pulsed electric fields and thermal processes.

Science.gov (United States)

Wu, Li; Zhao, Wei; Yang, Ruijin; Yan, Wenxu; Sun, Qianyan

2016-08-01

Pulsed electric field (PEF) processing is progressing towards application for liquid egg to ensure microbial safety. However, it usually causes protein aggregation, and the mechanism is still unclear. In this study, egg white protein was applied to investigate the changes in protein structure and mechanism of aggregates formation and a comparison was made with thermal treatment. Soluble protein content decreased with the increase of turbidity after both treatments. Fluorescence intensity and free sulfhydryl content were increased after being treated at 70 °C for 4 min. Less-remarkable changes of hydrophobicity were observed after PEF treatments (30 kV cm(-1) , 800 µs). Soluble and insoluble aggregates were observed by thermal treatment, and disulfide bonds were the main binding forces. The main components of insoluble aggregates formed by thermal treatment were ovotransferrin (30.58%), lysozyme (18.47%) and ovalbumin (14.20%). While only insoluble aggregates were detected during PEF processes, which consists of ovotransferrin (11.86%), lysozyme (21.11%) and ovalbumin (31.07%). Electrostatic interaction played a very important role in the aggregates formation. PEF had a minor impact on the structure of egg white protein. PEF had insignificant influence on heat-sensitive protein, indicating that PEF has potential in processing food with high biological activity and heat sensitive properties. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

Fault Detection and Isolation for Wind Turbine Electric Pitch System

DEFF Research Database (Denmark)

Zhu, Jiangsheng; Ma, Kuichao; Hajizadeh, Amin

2017-01-01

This paper presents a model-based fault detection and isolation scheme applied on electric pitch system of wind turbines. Pitch system is one of the most critical components due to its effect on the operational safety and the dynamics of wind turbines. Faults in this system should be precisely...... detected to prevent failures and decrease downtime. To detect faults of electric pitch actuators and sensors, an extended kalman filter (EKF) based multiple model adaptive estimation (MMAE) designed to estimate the states of the system. The proposed method is demonstrated in case studies. The simulation...

Direct Detection of Biotinylated Proteins by Mass Spectrometry

Science.gov (United States)

2015-01-01

Mass spectrometric strategies to identify protein subpopulations involved in specific biological functions rely on covalently tagging biotin to proteins using various chemical modification methods. The biotin tag is primarily used for enrichment of the targeted subpopulation for subsequent mass spectrometry (MS) analysis. A limitation of these strategies is that MS analysis does not easily discriminate unlabeled contaminants from the labeled protein subpopulation under study. To solve this problem, we developed a flexible method that only relies on direct MS detection of biotin-tagged proteins called “Direct Detection of Biotin-containing Tags” (DiDBiT). Compared with conventional targeted proteomic strategies, DiDBiT improves direct detection of biotinylated proteins ∼200 fold. We show that DiDBiT is applicable to several protein labeling protocols in cell culture and in vivo using cell permeable NHS-biotin and incorporation of the noncanonical amino acid, azidohomoalanine (AHA), into newly synthesized proteins, followed by click chemistry tagging with biotin. We demonstrate that DiDBiT improves the direct detection of biotin-tagged newly synthesized peptides more than 20-fold compared to conventional methods. With the increased sensitivity afforded by DiDBiT, we demonstrate the MS detection of newly synthesized proteins labeled in vivo in the rodent nervous system with unprecedented temporal resolution as short as 3 h. PMID:25117199

Protein detection using biobarcodes.

Science.gov (United States)

Müller, Uwe R

2006-10-01

Over the past 50 years the development of assays for the detection of protein analytes has been driven by continuing demands for higher levels of sensitivity and multiplexing. The result has been a progression of sandwich-type immunoassays, starting with simple radioisotopic, colorimetric, or fluorescent labeling systems to include various enzymatic or nanostructure-based signal amplification schemes, with a concomitant sensitivity increase of over 1 million fold. Multiplexing of samples and tests has been enabled by microplate and microarray platforms, respectively, or lately by various molecular barcoding systems. Two different platforms have emerged as the current front-runners by combining a nucleic acid amplification step with the standard two-sided immunoassay. In both, the captured protein analyte is replaced by a multiplicity of oligonucleotides that serve as surrogate targets. One of these platforms employs DNA or RNA polymerases for the amplification step, while detection is by fluorescence. The other is based on gold nanoparticles for both amplification as well as detection. The latter technology, now termed Biobarcode, is completely enzyme-free and offers potentially much higher multiplexing power.

Improved detection of calcium-binding proteins in polyacrylamide gels

International Nuclear Information System (INIS)

Anthony, F.A.; Babitch, J.A.

1984-01-01

The authors refined the method of Schibeci and Martonosi (1980) to enhance detection of calcium-binding proteins in polyacrylamide gels using 45 Ca 2+ . Their efforts have produced a method which is shorter, has 40-fold greater sensitivity over the previous method, and will detect 'EF hand'-containing calcium-binding proteins in polyacrylamide gels below the 0.5 μg level. In addition this method will detect at least one example from every described class of calcium-binding protein, including lectins and γ-carboxyglutamic acid containing calcium-binding proteins. The method should be useful for detecting calcium-binding proteins which may trigger neurotransmitter release. (Auth.)

Detection of weak electric fields by sharks, rays, and skates.

Science.gov (United States)

Adair, Robert K.; Astumian, R. Dean; Weaver, James C.

1998-09-01

The elasmobranchs-sharks, rays, and skates-can detect very weak electric fields in their aqueous environment through a complex sensory system, the ampullae of Lorenzini. The ampullae are conducting tubes that connect the surface of the animal to its interior. In the presence of an electric field, the potential of the surface of the animal will differ from that of the interior and that potential is applied across the apical membrane of the special sensory cells that line the ampullae. The firing rate of the afferent neurons that transmit signals from the ampullae has been shown to vary with that potential. We show that those firing rates can be described quantitatively in terms of synchronous firing of the sensory cells that feed the neurons. We demonstrate that such synchronism follows naturally from a hypothetical weak cell-to-cell interaction that results in a self-organization of the sensory cells. Moreover, the pulse rates of those cells-and the neurons that service the cells-can be expected to vary with the imposed electric fields in accord with measured values through actions of voltage gated transmembrane proteins in the apical sector of the cell membranes that admit Ca(++) ions. We also present a more conjectural model of signal processing at the neuron level that could exploit small differences in firing rates of nerve fibers servicing different ampullae to send an unambiguous signal to the central nervous system of the animal. (c) 1998 American Institute of Physics.

Immunofluorescence detection of pea protein in meat products.

Science.gov (United States)

Petrášová, Michaela; Pospiech, Matej; Tremlová, Bohuslava; Javůrková, Zdeňka

2016-08-01

In this study we developed an immunofluorescence method to detect pea protein in meat products. Pea protein has a high nutritional value but in sensitive individuals it may be responsible for causing allergic reactions. We produced model meat products with various additions of pea protein and flour; the detection limit (LOD) of the method for pea flour was 0.5% addition, and for pea protein it was 0.001% addition. The repeatabilities and reproducibilities for samples both positive and negative for pea protein were all 100%. In a blind test with model products and commercial samples, there was no statistically significant difference (p > 0.05) between the declared concentrations of pea protein and flour and the immunofluorescence method results. Sensitivity was 1.06 and specificity was 1.00. These results show that the immunofluorescence method is suitable for the detection of pea protein in meat products.

Rotation Detection Using the Precession of Molecular Electric Dipole Moment

Science.gov (United States)

Ke, Yi; Deng, Xiao-Bing; Hu, Zhong-Kun

2017-11-01

We present a method to detect the rotation by using the precession of molecular electric dipole moment in a static electric field. The molecular electric dipole moments are polarized under the static electric field and a nonzero electric polarization vector emerges in the molecular gas. A resonant radio-frequency pulse electric field is applied to realize a 90° flip of the electric polarization vector of a particular rotational state. After the pulse electric field, the electric polarization vector precesses under the static electric field. The rotation induces a shift in the precession frequency which is measured to deduce the angular velocity of the rotation. The fundamental sensitivity limit of this method is estimated. This work is only a proposal and does not involve experimental results.

Magnetogastrographic detection of gastric electrical response activity in humans

International Nuclear Information System (INIS)

Irimia, Andrei; Richards, William O; Bradshaw, L Alan

2006-01-01

The detection and characterization of gastric electrical activity has important clinical applications, including the early diagnosis of gastric diseases in humans. In mammals, this phenomenon has two important features: an electrical control activity (ECA) that manifests itself as an electric slow wave (with a frequency of 3 cycles per minute in humans) and an electrical response activity (ERA) that is characterized by spiking potentials during the plateau phase of the ECA. Whereas the ECA has been recorded in humans both invasively and non-invasively (magnetogastrography-MGG), the ERA has never been detected non-invasively in humans before. In this paper, we report on our progress towards the non-invasive detection of ERA from the human stomach using a procedure that involves the application of principal component analysis to MGG recordings, which were acquired in our case from ten normal human patients using a Superconducting QUantum Interference Device (SQUID) magnetometer. Both pre- and post-prandial recordings were acquired for each patient and 20 min of recordings (10 min of pre-prandial and 10 min of post-prandial data) were analysed for each patient. The mean percentage of ECA slow waves that were found to exhibit spikes of suspected ERA origin was 41% and 61% for pre- and post-prandial recordings, respectively, implying a 47% ERA increase post-prandially (P < 0.0001 at a 95% confidence level). The detection of ERA in humans is highly encouraging and points to the possible use of non-invasive ERA recordings as a valuable tool for the study of human gastric disorders

Signal amplification in electrochemical detection of buckwheat allergenic protein using field effect transistor biosensor by introduction of anionic surfactant

Directory of Open Access Journals (Sweden)

Sho Hideshima

2016-03-01

Full Text Available Food allergens, especially buckwheat proteins, sometimes induce anaphylactic shock in patients after ingestion. Development of a simple and rapid screening method based on a field effect transistor (FET biosensor for food allergens in food facilities or products is in demand. In this study, we achieved the FET detection of a buckwheat allergenic protein (BWp16, which is not charged enough to be electrically detected by FET biosensors, by introducing additional negative charges from anionic surfactants to the target proteins. A change in the FET characteristics reflecting surface potential caused by the adsorption of target charged proteins was observed when the target sample was coupled with the anionic surfactant (sodium dodecyl sulfate; SDS, while no significant response was detected without any surfactant treatment. It was suggested that the surfactant conjugated with the protein could be useful for the charge amplification of the target proteins. The surface plasmon resonance analysis revealed that the SDS-coupled proteins were successfully captured by the receptors immobilized on the sensing surface. Additionally, we obtained the FET responses at various concentrations of BWp16 ranging from 1 ng/mL to 10 μg/mL. These results suggest that a signal amplification method for FET biosensing is useful for allergen detection in the food industry. Keywords: Field effect transistor biosensor, Food allergen, Signal amplification, Ionic surfactant, Intrinsic charge

Comparison of membrane electroporation and protein denature in response to pulsed electric field with different durations.

Science.gov (United States)

Huang, Feiran; Fang, Zhihui; Mast, Jason; Chen, Wei

2013-05-01

In this paper, we compared the minimum potential differences in the electroporation of membrane lipid bilayers and the denaturation of membrane proteins in response to an intensive pulsed electric field with various pulse durations. Single skeletal muscle fibers were exposed to a pulsed external electric field. The field-induced changes in the membrane integrity (leakage current) and the Na channel currents were monitored to identify the minimum electric field needed to damage the membrane lipid bilayer and the membrane proteins, respectively. We found that in response to a relatively long pulsed electric shock (longer than the membrane intrinsic time constant), a lower membrane potential was needed to electroporate the cell membrane than for denaturing the membrane proteins, while for a short pulse a higher membrane potential was needed. In other words, phospholipid bilayers are more sensitive to the electric field than the membrane proteins for a long pulsed shock, while for a short pulse the proteins become more vulnerable. We can predict that for a short or ultrashort pulsed electric shock, the minimum membrane potential required to start to denature the protein functions in the cell plasma membrane is lower than that which starts to reduce the membrane integrity. Copyright © 2013 Wiley Periodicals, Inc.

Electrical Versus Optical: Comparing Methods for Detecting Terahertz Radiation Using Neon Lamps

Science.gov (United States)

Slocombe, L. L.; Lewis, R. A.

2018-05-01

Terahertz radiation impinging on a lit neon tube causes additional ionization of the encapsulated gas. As a result, the electrical current flowing between the electrodes increases and the glow discharge in the tube brightens. These dual phenomena suggest two distinct modes of terahertz sensing. The electrical mode simply involves measuring the electrical current. The optical mode involves monitoring the brightness of the weakly ionized plasma glow discharge. Here, we directly compare the two detection modes under identical experimental conditions. We measure 0.1-THz radiation modulated at frequencies in the range 0.1-10 kHz, for lamp currents in the range 1-10 mA. We find that electrical detection provides a superior signal-to-noise ratio while optical detection has a faster response. Either method serves as the basis of a compact, robust, and inexpensive room-temperature detector of terahertz radiation.

ELISA for Detection of Soya Proteins in Meat Products

Directory of Open Access Journals (Sweden)

Eva Renčová

2009-01-01

Full Text Available Indirect competitive ELISA method for the detection of soya proteins in meat products was developed. The detection limit of the method is 0.5% of the weight of added soya protein. A total of 131 meat product samples such as salamis or sausages from the Czech Republic market were investigated for the presence of soya proteins. Soya proteins were detected in 84% of the investigated samples without any declaration on the package of the product. The use of vegetable additives, namely soya in meat products in the market of the Czech Republic is very frequent and the restriction of its usage by legislation relates only to some kinds of durable products and ham (Act 264/2003 Coll.. The need for sensitive inspecting methods for soya protein detection is not only associated with the economic aspect (adulteration, but mainly with consumer health protection in case of allergy to soya proteins.

Protein permeation through an electrically tunable membrane

International Nuclear Information System (INIS)

Jou, Ining A; Melnikov, Dmitriy V; Gracheva, Maria E

2016-01-01

Protein filtration is important in many fields of science and technology such as medicine, biology, chemistry, and engineering. Recently, protein separation and filtering with nanoporous membranes has attracted interest due to the possibility of fast separation and high throughput volume. This, however, requires understanding of the protein’s dynamics inside and in the vicinity of the nanopore. In this work, we utilize a Brownian dynamics approach to study the motion of the model protein insulin in the membrane–electrolyte electrostatic potential. We compare the results of the atomic model of the protein with the results of a coarse-grained and a single-bead model, and find that the coarse-grained representation of protein strikes the best balance between the accuracy of the results and the computational effort required. Contrary to common belief, we find that to adequately describe the protein, a single-bead model cannot be utilized without a significant effort to tabulate the simulation parameters. Similar to results for nanoparticle dynamics, our findings also indicate that the electric field and the electro-osmotic flow due to the applied membrane and electrolyte biases affect the capture and translocation of the biomolecule by either attracting or repelling it to or from the nanopore. Our computational model can also be applied to other types of proteins and separation conditions. (paper)

Electrical detection of liquid lithium leaks from pipe joints

Energy Technology Data Exchange (ETDEWEB)

Schwartz, J. A., E-mail: jschwart@pppl.gov; Jaworski, M. A.; Mehl, J.; Kaita, R.; Mozulay, R. [Princeton Plasma Physics Laboratory, Princeton, New Jersey 08543-0451 (United States)

2014-11-15

A test stand for flowing liquid lithium is under construction at Princeton Plasma Physics Laboratory. As liquid lithium reacts with atmospheric gases and water, an electrical interlock system for detecting leaks and safely shutting down the apparatus has been constructed. A defense in depth strategy is taken to minimize the risk and impact of potential leaks. Each demountable joint is diagnosed with a cylindrical copper shell electrically isolated from the loop. By monitoring the electrical resistance between the pipe and the copper shell, a leak of (conductive) liquid lithium can be detected. Any resistance of less than 2 kΩ trips a relay, shutting off power to the heaters and pump. The system has been successfully tested with liquid gallium as a surrogate liquid metal. The circuit features an extensible number of channels to allow for future expansion of the loop. To ease diagnosis of faults, the status of each channel is shown with an analog front panel LED, and monitored and logged digitally by LabVIEW.

Electrical detection of spin current and spin relaxation in nonmagnetic semiconductors

International Nuclear Information System (INIS)

Miah, M Idrish

2008-01-01

We report an electrical method for the detection of spin current and spin relaxation in nonmagnetic semiconductors. Optically polarized spins are dragged by an electric field in GaAs. We use the anomalous Hall effect for the detection of spin current and spin relaxation. It is found that the effect depends on the electric field and doping density as well as on temperature, but not on the excitation power. A calculation for the effect is performed using the measured spin polarization by a pump-probe experiment. The results are also discussed in comparison with a quantitative evaluation of the spin lifetimes of the photogenerated electrons under drift in GaAs

Electrical detection of spin current and spin relaxation in nonmagnetic semiconductors

Energy Technology Data Exchange (ETDEWEB)

Miah, M Idrish [Nanoscale Science and Technology Centre and School of Biomolecular and Physical Sciences, Griffith University, Nathan, Brisbane, QLD 4111 (Australia); Department of Physics, University of Chittagong, Chittagong 4331 (Bangladesh)], E-mail: m.miah@griffith.edu.au

2008-09-21

We report an electrical method for the detection of spin current and spin relaxation in nonmagnetic semiconductors. Optically polarized spins are dragged by an electric field in GaAs. We use the anomalous Hall effect for the detection of spin current and spin relaxation. It is found that the effect depends on the electric field and doping density as well as on temperature, but not on the excitation power. A calculation for the effect is performed using the measured spin polarization by a pump-probe experiment. The results are also discussed in comparison with a quantitative evaluation of the spin lifetimes of the photogenerated electrons under drift in GaAs.

Solid-state NMR analysis of membrane proteins and protein aggregates by proton detected spectroscopy

International Nuclear Information System (INIS)

Zhou, Donghua H.; Nieuwkoop, Andrew J.; Berthold, Deborah A.; Comellas, Gemma; Sperling, Lindsay J.; Tang, Ming; Shah, Gautam J.; Brea, Elliott J.; Lemkau, Luisel R.; Rienstra, Chad M.

2012-01-01

Solid-state NMR has emerged as an important tool for structural biology and chemistry, capable of solving atomic-resolution structures for proteins in membrane-bound and aggregated states. Proton detection methods have been recently realized under fast magic-angle spinning conditions, providing large sensitivity enhancements for efficient examination of uniformly labeled proteins. The first and often most challenging step of protein structure determination by NMR is the site-specific resonance assignment. Here we demonstrate resonance assignments based on high-sensitivity proton-detected three-dimensional experiments for samples of different physical states, including a fully-protonated small protein (GB1, 6 kDa), a deuterated microcrystalline protein (DsbA, 21 kDa), a membrane protein (DsbB, 20 kDa) prepared in a lipid environment, and the extended core of a fibrillar protein (α-synuclein, 14 kDa). In our implementation of these experiments, including CONH, CO(CA)NH, CANH, CA(CO)NH, CBCANH, and CBCA(CO)NH, dipolar-based polarization transfer methods have been chosen for optimal efficiency for relatively high protonation levels (full protonation or 100 % amide proton), fast magic-angle spinning conditions (40 kHz) and moderate proton decoupling power levels. Each H–N pair correlates exclusively to either intra- or inter-residue carbons, but not both, to maximize spectral resolution. Experiment time can be reduced by at least a factor of 10 by using proton detection in comparison to carbon detection. These high-sensitivity experiments are especially important for membrane proteins, which often have rather low expression yield. Proton-detection based experiments are expected to play an important role in accelerating protein structure elucidation by solid-state NMR with the improved sensitivity and resolution.

Network single-walled carbon nanotube biosensors for fast and highly sensitive detection of proteins

International Nuclear Information System (INIS)

Hu Pingan; Zhang Jia; Wen Zhenzhong; Zhang Can

2011-01-01

Detection of proteins is powerfully assayed in the diagnosis of diseases. A strategy for the development of an ultrahigh sensitivity biosensor based on a network single-walled carbon nanotube (SWNT) field-effect transistor (FET) has been demonstrated. Metallic SWNTs (m-SWNTs) in the network nanotube FET were selectively removed or cut via a carefully controlled procedure of electrical break-down (BD), and left non-conducting m-SWNTs which magnified the Schottky barrier (SB) area. This nanotube FET exhibited ultrahigh sensitivity and fast response to biomolecules. The lowest detection limit of 0.5 pM was achieved by exploiting streptavidin (SA) or a biotin/SA pair as the research model, and BD-treated nanotube biosensors had a 2 x 10 4 -fold lower minimum detectable concentration than the device without BD treatment. The response time is in the range of 0.3-3 min.

Neuromuscular electrical stimulation prior to presleep protein feeding stimulates the use of protein-derived amino acids for overnight muscle protein synthesis.

Science.gov (United States)

Dirks, Marlou L; Groen, Bart B L; Franssen, Rinske; van Kranenburg, Janneau; van Loon, Luc J C

2017-01-01

Short periods of muscle disuse result in substantial skeletal muscle atrophy. Recently, we showed that both neuromuscular electrical stimulation (NMES) as well as presleep dietary protein ingestion represent effective strategies to stimulate muscle protein synthesis rates. In this study, we test our hypothesis that NMES can augment the use of presleep protein-derived amino acids for overnight muscle protein synthesis in older men. Twenty healthy, older [69 ± 1 (SE) yr] men were subjected to 24 h of bed rest, starting at 8:00 AM. In the evening, volunteers were subjected to 70-min 1-legged NMES, while the other leg served as nonstimulated control (CON). Immediately following NMES, 40 g of intrinsically l-[1- 13 C]-phenylalanine labeled protein was ingested prior to sleep. Blood samples were taken throughout the night, and muscle biopsies were obtained from both legs in the evening and the following morning (8 h after protein ingestion) to assess dietary protein-derived l-[1- 13 C]-phenylalanine enrichments in myofibrillar protein. Plasma phenylalanine concentrations and plasma l-[1- 13 C]-phenylalanine enrichments increased significantly following protein ingestion and remained elevated for up to 6 h after protein ingestion (P protein-bound l-[1- 13 C]-phenylalanine enrichments (MPE) increased to a greater extent in the stimulated compared with the control leg (0.0344 ± 0.0019 vs. 0.0297 ± 0.0016 MPE, respectively; P protein-derived amino acids in the NMES compared with CON leg. In conclusion, application of NMES prior to presleep protein feeding stimulates the use of dietary protein-derived amino acids for overnight muscle protein synthesis in older men. Neuromuscular electrical stimulation (NMES) as well as presleep dietary protein ingestion represent effective strategies to stimulate muscle protein synthesis rates. Here we demonstrate that in older men after a day of bed rest, the application of NMES prior to presleep protein feeding stimulates the use of

Fluorescence-Based Multiplex Protein Detection Using Optically Encoded Microbeads

Directory of Open Access Journals (Sweden)

Dae Hong Jeong

2012-03-01

Full Text Available Potential utilization of proteins for early detection and diagnosis of various diseases has drawn considerable interest in the development of protein-based multiplex detection techniques. Among the various techniques for high-throughput protein screening, optically-encoded beads combined with fluorescence-based target monitoring have great advantages over the planar array-based multiplexing assays. This review discusses recent developments of analytical methods of screening protein molecules on microbead-based platforms. These include various strategies such as barcoded microbeads, molecular beacon-based techniques, and surface-enhanced Raman scattering-based techniques. Their applications for label-free protein detection are also addressed. Especially, the optically-encoded beads such as multilayer fluorescence beads and SERS-encoded beads are successful for generating a large number of coding.

A highly sensitive method for detection of molybdenum-containing proteins

International Nuclear Information System (INIS)

Kalakutskii, K.L.; Shvetsov, A.A.; Bursakov, S.A.; Letarov, A.V.; Zabolotnyi, A.I.; L'vov, N.P.

1992-01-01

A highly sensitive method for detection of molybdenum-containing proteins in gels after electrophoresis has been developed. The method involves in vitro labeling of the proteins with the radioactive isotope 185 W. The method used to detect molybdenum-accumulating proteins in lupine seeds, xanthine dehydrogenase and another molybdenum-containing protein in wheat, barley, and pea seedlings, and nitrate reductase and xanthine dehydrogenase in bacteroides from lupine nodules. Nitrogenase could not be detected by the method. 16 refs., 5 figs

Detection of electric field around field-reversed configuration plasma

International Nuclear Information System (INIS)

Ikeyama, Taeko; Hiroi, Masanori; Nogi, Yasuyuki; Ohkuma, Yasunori

2010-01-01

Electric-field probes consisting of copper plates are developed to measure electric fields in a vacuum region around a plasma. The probes detect oscillating electric fields with a maximum strength of approximately 100 V/m through a discharge. Reproducible signals from the probes are obtained with an unstable phase dominated by a rotational instability. It is found that the azimuthal structure of the electric field can be explained by the sum of an n=2 mode charge distribution and a convex-surface electron distribution on the deformed separatrix at the unstable phase. The former distribution agrees with that anticipated from the diamagnetic drift motions of plasma when the rotational instability occurs. The latter distribution suggests that an electron-rich plasma covers the separatrix.

Protein lipoxidation: Detection strategies and challenges

Directory of Open Access Journals (Sweden)

Giancarlo Aldini

2015-08-01

Full Text Available Enzymatic and non-enzymatic lipid metabolism can give rise to reactive species that may covalently modify cellular or plasma proteins through a process known as lipoxidation. Under basal conditions, protein lipoxidation can contribute to normal cell homeostasis and participate in signaling or adaptive mechanisms, as exemplified by lipoxidation of Ras proteins or of the cytoskeletal protein vimentin, both of which behave as sensors of electrophilic species. Nevertheless, increased lipoxidation under pathological conditions may lead to deleterious effects on protein structure or aggregation. This can result in impaired degradation and accumulation of abnormally folded proteins contributing to pathophysiology, as may occur in neurodegenerative diseases. Identification of the protein targets of lipoxidation and its functional consequences under pathophysiological situations can unveil the modification patterns associated with the various outcomes, as well as preventive strategies or potential therapeutic targets. Given the wide structural variability of lipid moieties involved in lipoxidation, highly sensitive and specific methods for its detection are required. Derivatization of reactive carbonyl species is instrumental in the detection of adducts retaining carbonyl groups. In addition, use of tagged derivatives of electrophilic lipids enables enrichment of lipoxidized proteins or peptides. Ultimate confirmation of lipoxidation requires high resolution mass spectrometry approaches to unequivocally identify the adduct and the targeted residue. Moreover, rigorous validation of the targets identified and assessment of the functional consequences of these modifications are essential. Here we present an update on methods to approach the complex field of lipoxidation along with validation strategies and functional assays illustrated with well-studied lipoxidation targets.

The Architecture Design of Detection and Calibration System for High-voltage Electrical Equipment

Science.gov (United States)

Ma, Y.; Lin, Y.; Yang, Y.; Gu, Ch; Yang, F.; Zou, L. D.

2018-01-01

With the construction of Material Quality Inspection Center of Shandong electric power company, Electric Power Research Institute takes on more jobs on quality analysis and laboratory calibration for high-voltage electrical equipment, and informationization construction becomes urgent. In the paper we design a consolidated system, which implements the electronic management and online automation process for material sampling, test apparatus detection and field test. In the three jobs we use QR code scanning, online Word editing and electronic signature. These techniques simplify the complex process of warehouse management and testing report transferring, and largely reduce the manual procedure. The construction of the standardized detection information platform realizes the integrated management of high-voltage electrical equipment from their networking, running to periodic detection. According to system operation evaluation, the speed of transferring report is doubled, and querying data is also easier and faster.

Methods for detecting and locating leaks in containment facilities using electrical potential data and electrical resistance tomographic imaging techniques

Science.gov (United States)

Daily, William D.; Laine, Daren L.; Laine, Edwin F.

1997-01-01

Methods are provided for detecting and locating leaks in liners used as barriers in the construction of landfills, surface impoundments, water reservoirs, tanks, and the like. Electrodes are placed in the ground around the periphery of the facility, in the leak detection zone located between two liners if present, and/or within the containment facility. Electrical resistivity data is collected using these electrodes. This data is used to map the electrical resistivity distribution beneath the containment liner between two liners in a double-lined facility. In an alternative embodiment, an electrode placed within the lined facility is driven to an electrical potential with respect to another electrode placed at a distance from the lined facility (mise-a-la-masse). Voltage differences are then measured between various combinations of additional electrodes placed in the soil on the periphery of the facility, the leak detection zone, or within the facility. A leak of liquid though the liner material will result in an electrical potential distribution that can be measured at the electrodes. The leak position is located by determining the coordinates of an electrical current source pole that best fits the measured potentials with the constraints of the known or assumed resistivity distribution.

Immunofluorescence detection of milk protein in meat products

Directory of Open Access Journals (Sweden)

Michaela Petrášová

2015-05-01

Full Text Available Nowadays there are various vegetable protein additives intended for the manufacture of meat products in the food industry. These ingredients include both, plant-origin as well as animal-origin proteins. The most common vegetable additives include various types of flour, starch, fiber and plant protein. Among animal proteins, the most commonly used are plasma, collagen or milk protein. Milk protein is added to meat products due to its functional properties, such as emulsifying fats, improving the holding capacity of meat, improving juiciness, gel-forming capacity and affecting the taste of the product. Usage of these proteins, however, is currently limited by the effective legislation, not only in order to prevent consumer deception, but also because of their potential impact on consumers' health of. Thus, this issue has received considerable attention not only in the Czech Republic, but also globally. The main risk is the impossibility of selecting a suitable foodstuff for individuals with potential allergic reactions. The only option for allergic consumers to protect themselves is to strictly exclude the given allergen from their diet. Although the number of studies dealing with the reduction or loss of allergenicity is increasing, yet these practices are not common. Most of the population suffering from food allergies is thus still dependent on strict exclusion of foodstuffs causing adverse allergic reactions from their diet. Detection of allergens in foodstuffs is unfortunately quite difficult due to the fact that they occur in trace amounts and are often masked by different parts of the foodstuff. This research dealt with the detection of milk protein in meat products purchased in the market network of the Czech Republic, whereas declaration given by the manufacturer on the packaging for the small meat products purchased from the market was used to verify the detection of milk protein by the immunofluorescence method. 20 products were

Laser-based optical activity detection of amino acids and proteins

Energy Technology Data Exchange (ETDEWEB)

Reitsma, B.H.

1987-08-01

The optical activity detector (OAD) for HPLC is a selective detector for optically active substances including amino acids and proteins. Four free amino acids were resolved using cation-exchange chromatography followed by detection with refractive index detector (RI) for proline and threonine and the OAD to an ultraviolet absorbance detector (uv) for tyrosine and phenylalanine. Amino acid detection by refractive index is not sensitive and uv absorbance detects only three amino acids. Derivatization of amino acids to make them detectable by uv absorbance enhances the applicability of OA/uv for the determination of enantiomeric ratios. The separation of 16 dansyl-L-amino acids by RP-HPLC with detection by OA/uv is illustrated. Calculation of the specific rotation of 22 dansyl-L-amino acids shows that derivatization enhances the OA detectability of some amino acids but degrades that of others. RP-HPLC of proteins is a rapidly developing technique. Several researchers have reported the detection of multiple peaks when a pure protein is subjected to HPLC under certain conditions. These multiple peaks have been determined to be different conformations of the same protein. Since proteins are optically active, OA is a suitable detector. The RP-HPLC separation of conformers of soybean trypsin inhibitor is illustrated. Detection by OA/uv provides insights from the chromatogram unavailable from uv absorbance detection alone. In addition, identification of impurities is simplified with OA/uv. Specific rotations of the separated protein fractions show no significant change accompanying change in conformation. 163 refs., 13 figs., 9 tabs.

CMsearch: simultaneous exploration of protein sequence space and structure space improves not only protein homology detection but also protein structure prediction

KAUST Repository

Cui, Xuefeng

2016-06-15

Motivation: Protein homology detection, a fundamental problem in computational biology, is an indispensable step toward predicting protein structures and understanding protein functions. Despite the advances in recent decades on sequence alignment, threading and alignment-free methods, protein homology detection remains a challenging open problem. Recently, network methods that try to find transitive paths in the protein structure space demonstrate the importance of incorporating network information of the structure space. Yet, current methods merge the sequence space and the structure space into a single space, and thus introduce inconsistency in combining different sources of information. Method: We present a novel network-based protein homology detection method, CMsearch, based on cross-modal learning. Instead of exploring a single network built from the mixture of sequence and structure space information, CMsearch builds two separate networks to represent the sequence space and the structure space. It then learns sequence–structure correlation by simultaneously taking sequence information, structure information, sequence space information and structure space information into consideration. Results: We tested CMsearch on two challenging tasks, protein homology detection and protein structure prediction, by querying all 8332 PDB40 proteins. Our results demonstrate that CMsearch is insensitive to the similarity metrics used to define the sequence and the structure spaces. By using HMM–HMM alignment as the sequence similarity metric, CMsearch clearly outperforms state-of-the-art homology detection methods and the CASP-winning template-based protein structure prediction methods.

Electrically Excited Plasmonic Nanoruler for Biomolecule Detection.

Science.gov (United States)

Dathe, André; Ziegler, Mario; Hübner, Uwe; Fritzsche, Wolfgang; Stranik, Ondrej

2016-09-14

Plasmon-based sensors are excellent tools for a label-free detection of small biomolecules. An interesting group of such sensors are plasmonic nanorulers that rely on the plasmon hybridization upon modification of their morphology to sense nanoscale distances. Sensor geometries based on the interaction of plasmons in a flat metallic layer together with metal nanoparticles inherit unique advantages but need a special optical excitation configuration that is not easy to miniaturize. Herein, we introduce the concept of nanoruler excitation by direct, electrically induced generation of surface plasmons based on the quantum shot noise of tunneling currents. An electron tunneling junction consisting of a metal-dielectric-semiconductor heterostructure is directly incorporated into the nanoruler basic geometry. With the application of voltage on this modified nanoruler, the plasmon modes are directly excited without any additional optical component as a light source. We demonstrate via several experiments that this electrically driven nanoruler possesses similar properties as an optically exited one and confirm its sensing capabilities by the detection of the binding of small biomolecules such as antibodies. This new sensing principle could open the way to a new platform of highly miniaturized, integrated plasmonic sensors compatible with monolithic integrated circuits.

Electrical excitation and optical detection of ultrasounds in PZT based piezoelectric transducers

Energy Technology Data Exchange (ETDEWEB)

Babilotte, P; Diallo, O; Hue, L-P Tran Hu; Feuillard, G [University Francois Rabelais de Tours, Laboratory Imaging and Brain, Team Ultrasonic Characterisation and Piezoelectricity, ENIVL, Rue de la Chocolaterie, 41034 BLOIS CEDEX (France); Kosec, M; Kuscer, D, E-mail: philippe.babilotte@univ-tours.fr [Josef Stefan Institute, Jamova cesta 39, 1000 LJUBLJANA (Slovenia)

2011-01-01

The displacement response of piezoelectric PZT thick films fabricated by means of electrophoretic deposition and laid down an alumina substrate is investigated using coherent optical detection. According to thickness properties determined by electrical impedance measurements, the film presents a resonance around 40 MHz. Other resonance peaks are observed that correspond to eigen modes of the film substrate couple structure. Uniformity of the response of the integrated structure is studied across the surface of the sample when excited by either a continuous or impulse electrical voltage. Results on the amplitude of the detected signal versus the frequency and the input excitation voltage are reported. The optical detection used in these experiments is complementary to conventional techniques of characterization of piezoelectric devices such as electrical impedance measurements and allows getting information on the displacement response of the device.

Mass spectrometric detection of proteins in non-aqueous media : the case of prion proteins in biodiesel

Energy Technology Data Exchange (ETDEWEB)

Douma, M.D.; Kerr, G.M.; Brown, R.S.; Keller, B.O.; Oleschuk, R.D. [Queen' s Univ., Kingston, ON (Canada). Dept. of Chemistry

2008-08-15

This paper presented a filtration method for detecting protein traces in non-aqueous media. The extraction technique used a mixture of acetonitrile, non-ionic detergent and water along with filter disks with embedded C{sub 8}-modified silica particles to capture the proteins from non-aqueous samples. The extraction process was then followed by an elution of the protein from the filter disk and direct mass spectrometric detection and tryptic digestion with peptide mapping and MS/MS fragmentation of protein-specific peptides. The method was used to detect prion proteins in spiked biodiesel samples. A tryptic peptide with the sequence YGQGSPGGNR was used for unambiguous identification. Results of the study showed that the method is suitable for the large-scale testing of protein impurities in tallow-based biodiesel production processes. 33 refs., 6 figs.

Detection of significant protein coevolution.

Science.gov (United States)

Ochoa, David; Juan, David; Valencia, Alfonso; Pazos, Florencio

2015-07-01

The evolution of proteins cannot be fully understood without taking into account the coevolutionary linkages entangling them. From a practical point of view, coevolution between protein families has been used as a way of detecting protein interactions and functional relationships from genomic information. The most common approach to inferring protein coevolution involves the quantification of phylogenetic tree similarity using a family of methodologies termed mirrortree. In spite of their success, a fundamental problem of these approaches is the lack of an adequate statistical framework to assess the significance of a given coevolutionary score (tree similarity). As a consequence, a number of ad hoc filters and arbitrary thresholds are required in an attempt to obtain a final set of confident coevolutionary signals. In this work, we developed a method for associating confidence estimators (P values) to the tree-similarity scores, using a null model specifically designed for the tree comparison problem. We show how this approach largely improves the quality and coverage (number of pairs that can be evaluated) of the detected coevolution in all the stages of the mirrortree workflow, independently of the starting genomic information. This not only leads to a better understanding of protein coevolution and its biological implications, but also to obtain a highly reliable and comprehensive network of predicted interactions, as well as information on the substructure of macromolecular complexes using only genomic information. The software and datasets used in this work are freely available at: http://csbg.cnb.csic.es/pMT/. pazos@cnb.csic.es Supplementary data are available at Bioinformatics online. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

Hydrophobicity-driven self-assembly of protein and silver nanoparticles for protein detection using surface-enhanced Raman scattering.

Science.gov (United States)

Kahraman, Mehmet; Balz, Ben N; Wachsmann-Hogiu, Sebastian

2013-05-21

Surface-enhanced Raman scattering (SERS) is a promising analytical technique for the detection and characterization of biological molecules and structures. The role of hydrophobic and hydrophilic surfaces in the self-assembly of protein-metallic nanoparticle structures for label-free protein detection is demonstrated. Aggregation is driven by both the hydrophobicity of the surface as well as the charge of the proteins. The best conditions for obtaining a reproducible SERS signal that allows for sensitive, label-free protein detection are provided by the use of hydrophobic surfaces and 16 × 10(11) NPs per mL. A detection limit of approximately 0.5 μg mL(-1) is achieved regardless of the proteins' charge properties and size. The developed method is simple and can be used for reproducible and sensitive detection and characterization of a wide variety of biological molecules and various structures with different sizes and charge status.

Effect of electric charge on the transperitoneal transport of plasma proteins during CAPD

NARCIS (Netherlands)

Buis, B.; Koomen, G. C.; Imholz, A. L.; Struijk, D. G.; Reddingius, R. E.; Arisz, L.; Krediet, R. T.

1996-01-01

BACKGROUND: Controversy exists as to whether electric charges of plasma proteins influence their transport across the peritoneal membrane during CAPD. Fixed negative charges in the peritoneal membrane are diminished during peritonitis in rats. METHODS: Peritoneal clearances of 10 proteins and their

DiffSLC: A graph centrality method to detect essential proteins of a protein-protein interaction network.

Science.gov (United States)

Mistry, Divya; Wise, Roger P; Dickerson, Julie A

2017-01-01

Identification of central genes and proteins in biomolecular networks provides credible candidates for pathway analysis, functional analysis, and essentiality prediction. The DiffSLC centrality measure predicts central and essential genes and proteins using a protein-protein interaction network. Network centrality measures prioritize nodes and edges based on their importance to the network topology. These measures helped identify critical genes and proteins in biomolecular networks. The proposed centrality measure, DiffSLC, combines the number of interactions of a protein and the gene coexpression values of genes from which those proteins were translated, as a weighting factor to bias the identification of essential proteins in a protein interaction network. Potentially essential proteins with low node degree are promoted through eigenvector centrality. Thus, the gene coexpression values are used in conjunction with the eigenvector of the network's adjacency matrix and edge clustering coefficient to improve essentiality prediction. The outcome of this prediction is shown using three variations: (1) inclusion or exclusion of gene co-expression data, (2) impact of different coexpression measures, and (3) impact of different gene expression data sets. For a total of seven networks, DiffSLC is compared to other centrality measures using Saccharomyces cerevisiae protein interaction networks and gene expression data. Comparisons are also performed for the top ranked proteins against the known essential genes from the Saccharomyces Gene Deletion Project, which show that DiffSLC detects more essential proteins and has a higher area under the ROC curve than other compared methods. This makes DiffSLC a stronger alternative to other centrality methods for detecting essential genes using a protein-protein interaction network that obeys centrality-lethality principle. DiffSLC is implemented using the igraph package in R, and networkx package in Python. The python package can be

Electrical detection of magnetization dynamics via spin rectification effects

Energy Technology Data Exchange (ETDEWEB)

Harder, Michael, E-mail: michael.harder@umanitoba.ca; Gui, Yongsheng, E-mail: ysgui@physics.umanitoba.ca; Hu, Can-Ming, E-mail: hu@physics.umanitoba.ca

2016-11-23

The purpose of this article is to review the current status of a frontier in dynamic spintronics and contemporary magnetism, in which much progress has been made in the past decade, based on the creation of a variety of micro and nanostructured devices that enable electrical detection of magnetization dynamics. The primary focus is on the physics of spin rectification effects, which are well suited for studying magnetization dynamics and spin transport in a variety of magnetic materials and spintronic devices. Intended to be intelligible to a broad audience, the paper begins with a pedagogical introduction, comparing the methods of electrical detection of charge and spin dynamics in semiconductors and magnetic materials respectively. After that it provides a comprehensive account of the theoretical study of both the angular dependence and line shape of electrically detected ferromagnetic resonance (FMR), which is summarized in a handbook format easy to be used for analysing experimental data. We then review and examine the similarity and differences of various spin rectification effects found in ferromagnetic films, magnetic bilayers and magnetic tunnel junctions, including a discussion of how to properly distinguish spin rectification from the spin pumping/inverse spin Hall effect generated voltage. After this we review the broad applications of rectification effects for studying spin waves, nonlinear dynamics, domain wall dynamics, spin current, and microwave imaging. We also discuss spin rectification in ferromagnetic semiconductors. The paper concludes with both historical and future perspectives, by summarizing and comparing three generations of FMR spectroscopy which have been developed for studying magnetization dynamics.

Effect of transcutaneous electrical muscle stimulation on postoperative muscle mass and protein synthesis

DEFF Research Database (Denmark)

Vinge, O; Edvardsen, L; Jensen, F

1996-01-01

In an experimental study, 13 patients undergoing major elective abdominal surgery were given postoperative transcutaneous electrical muscle stimulation (TEMS) to the quadriceps femoris muscle on one leg; the opposite leg served as control. Changes in cross-sectional area (CSA) and muscle protein ...... protein synthesis and muscle mass after abdominal surgery and should be evaluated in other catabolic states with muscle wasting.......In an experimental study, 13 patients undergoing major elective abdominal surgery were given postoperative transcutaneous electrical muscle stimulation (TEMS) to the quadriceps femoris muscle on one leg; the opposite leg served as control. Changes in cross-sectional area (CSA) and muscle protein...... synthesis were assessed by computed tomography and ribosome analysis of percutaneous muscle biopsies before surgery and on the sixth postoperative day. The percentage of polyribosomes in the ribosome suspension decreased significantly (P

Measurement of absolute displacement-amplitude of ultrasonic wave using piezo-electric detection method

Energy Technology Data Exchange (ETDEWEB)

Park, Seong Hyun; Kim, Jong Beom; Jhang, Kyung Young [Hanyang University, Seoul (Korea, Republic of)

2017-02-15

A nonlinear ultrasonic parameter is defined by the ratio of displacement amplitude of the fundamental frequency component to that of the second-order harmonic frequency component. In this study, the ultrasonic displacement amplitude of an SUS316 specimen was measured via a piezo-electric-based method to identify the validity of piezo-electric detection method. For comparison, the ultrasonic displacement was also determined via a laser-based Fabry-Pérot interferometer. The experimental results for both measurements were in good agreement. Additionally, the stability of the repeated test results from the piezo-electric method exceeded that of the laser-interferometric method. This result indicated that the piezo-electric detection method can be utilized to measure a nonlinear ultrasonic parameter due to its excellent stability although it involves a complicated process.

Measurement of absolute displacement-amplitude of ultrasonic wave using piezo-electric detection method

International Nuclear Information System (INIS)

Park, Seong Hyun; Kim, Jong Beom; Jhang, Kyung Young

2017-01-01

A nonlinear ultrasonic parameter is defined by the ratio of displacement amplitude of the fundamental frequency component to that of the second-order harmonic frequency component. In this study, the ultrasonic displacement amplitude of an SUS316 specimen was measured via a piezo-electric-based method to identify the validity of piezo-electric detection method. For comparison, the ultrasonic displacement was also determined via a laser-based Fabry-Pérot interferometer. The experimental results for both measurements were in good agreement. Additionally, the stability of the repeated test results from the piezo-electric method exceeded that of the laser-interferometric method. This result indicated that the piezo-electric detection method can be utilized to measure a nonlinear ultrasonic parameter due to its excellent stability although it involves a complicated process

Non-invasive electrocardiogram detection of in vivo zebrafish embryos using electric potential sensors

Science.gov (United States)

Rendon-Morales, E.; Prance, R. J.; Prance, H.; Aviles-Espinosa, R.

2015-11-01

In this letter, we report the continuous detection of the cardiac electrical activity in embryonic zebrafish using a non-invasive approach. We present a portable and cost-effective platform based on the electric potential sensing technology, to monitor in vivo electrocardiogram activity from the zebrafish heart. This proof of principle demonstration shows how electrocardiogram measurements from the embryonic zebrafish may become accessible by using electric field detection. We present preliminary results using the prototype, which enables the acquisition of electrophysiological signals from in vivo 3 and 5 days-post-fertilization zebrafish embryos. The recorded waveforms show electrocardiogram traces including detailed features such as QRS complex, P and T waves.

Protein Detection with Aptamer Biosensors

Directory of Open Access Journals (Sweden)

Regina Stoltenburg

2008-07-01

Full Text Available Aptamers have been developed for different applications. Their use as new biological recognition elements in biosensors promises progress for fast and easy detection of proteins. This new generation of biosensor (aptasensors will be more stable and well adapted to the conditions of real samples because of the specific properties of aptamers.

Detection of heart-type fatty acid-binding protein (h-FABP) using piezoresistive polymer microcantilevers functionalized by a dry method

Science.gov (United States)

Agarwal, Dilip Kumar; Prasad, Abhinav; Vinchurkar, Madhuri; Gandhi, Sahir; Prabhakar, Deepika; Mukherji, Soumyo; Rao, V. Ramgopal

2018-03-01

Piezoresistive microcantilever-based sensor platform is being used for the last two decades due to their low cost, rapid response and label-free detection system. In this work, we are reporting a microfabricated piezoresistive SU-8/carbon black (polymer cantilever)-based sensor platform for the detection of a clinically important early-stage cardiac marker, i.e., fatty acid-binding protein. It is a most preferred cardiac marker for the diagnosis of acute myocardial infarction. The embodiment of the sensor is a SU-8 microcantilever chip with an integrated nanoparticle composite (carbon black) as a piezoresistor for on-chip electrical transduction. Prior to improving the sensing and susceptibility towards the specific target biomolecule (i.e., h-FABP), the fabricated SU-8 polymer cantilevers were subjected to tailored functionalization. This includes the use of an in-house dry method of hot wire chemical vapour deposition technique to graft amine groups onto the SU-8 surface. The surface-modified microcantilevers were further integrated with a polydimethylsiloxane liquid flow cell and connected externally with an electrical read-out system. Immobilization of the antibody corresponding to the marker protein on the microcantilever surface and subsequent recording of the signal generated upon the antibody-antigen interaction were carried out inside the liquid flow cell. Using our optimized immobilization protocol with this experimental set-up, we were successfully able to detect h-FABP concentration as low as 100 ng/ml.

Highly sensitive detection for proteins using graphene oxide-aptamer based sensors.

Science.gov (United States)

Gao, Li; Li, Qin; Li, Raoqi; Yan, Lirong; Zhou, Yang; Chen, Keping; Shi, Haixia

2015-07-07

In recent years, the detection of proteins by using bare graphene oxide (GO) to quench the fluorescence of fluorescein-labeled aptamers has been reported. However, the proteins can be adsorbed on the surface of bare GO to prevent the sensitivity from further being improved. In order to solve this problem, polyethylene glycol (PEG)-protected GO was used to prevent the proteins using thrombin as an example from nonspecific binding. The detection limit was improved compared to bare GO under the optimized ratio of GO to PEG concentration. The results show that our method is a promising technique for the detection of proteins.

Administering and Detecting Protein Marks on Arthropods for Dispersal Research.

Science.gov (United States)

Hagler, James R; Machtley, Scott A

2016-01-28

Monitoring arthropod movement is often required to better understand associated population dynamics, dispersal patterns, host plant preferences, and other ecological interactions. Arthropods are usually tracked in nature by tagging them with a unique mark and then re-collecting them over time and space to determine their dispersal capabilities. In addition to actual physical tags, such as colored dust or paint, various types of proteins have proven very effective for marking arthropods for ecological research. Proteins can be administered internally and/or externally. The proteins can then be detected on recaptured arthropods with a protein-specific enzyme-linked immunosorbent assay (ELISA). Here we describe protocols for externally and internally tagging arthropods with protein. Two simple experimental examples are demonstrated: (1) an internal protein mark introduced to an insect by providing a protein-enriched diet and (2) an external protein mark topically applied to an insect using a medical nebulizer. We then relate a step-by-step guide of the sandwich and indirect ELISA methods used to detect protein marks on the insects. In this demonstration, various aspects of the acquisition and detection of protein markers on arthropods for mark-release-recapture, mark-capture, and self-mark-capture types of research are discussed, along with the various ways that the immunomarking procedure has been adapted to suit a wide variety of research objectives.

Studies on Some Biophysical Properties of the Serum Protein of Mice blood exposed to an electric field

International Nuclear Information System (INIS)

Hanafy, M.S.

2005-01-01

As an indication of the effect of the electric field on each of the dielectric properties and the molecular structure of the serum protein of the mice blood, an electric field of a 6 kv/m strength and 50 Hz frequency was directed to three groups of mice for exposure periods 30, 45 and 60 days respectively, and investigated directly. Another group was exposed to also 60 days, but investigated after 30 days from switching off the electric field for delayed effect studies. The molecular structure of the serum protein was studied by measuring each of the dielectric relaxation and the electric conductivity in the frequency range 0.15 MHz at 4 ± 0.5 degree C and the dielectric increment (Δ), relaxation time (τ) and average molecular radii (τ) were calculated for all groups. The absorption spectra of the extracted protein were also measured in the wavelength range 200 600 nm. Moreover, electrophoresis of enzymes B-esterase, lactate and Malate dehydrogenase extracted from the blood serum of exposed mice were taken by using the gel electrophoresis technique. The results indicated that exposure of the animals to 50 H, 6 kv/m electric field resulted in the decrease of serum protein permittivity values and increase its conductivity a fact that indicates pronounced changes in the molecular structure of total serum protein the exposed mice. In addition, the intensity of the absorption spectral bands of serum protein of exposed mice were found to decrease relative to unexposed mice. Also the enzymes B-esterase and lactate dehydrogenase were slightly affected by exposing to the electric field whereas their number of bands and their intensities changed relative to the unexposed mice but the malate dehydrogenase was not affected

Effect of processing on the detectability of peanut protein by ELISA.

Science.gov (United States)

Iqbal, Amjad; Ateeq, Nadia

2013-12-01

Chicken IgY was used for the detection and quantification of peanut proteins by indirect competitive ELISA. The method was optimized by using a checker board approach to determine the optimal concentration of coating antigen, primary antibody and secondary antibody. Peanut protein could be detected in foods down to levels of 10 ppm. The effect of physical (heat treatment at 80 °C and 100 °C) and chemical (acid, alkali and reducing sugar) treatments on the IgY binding of peanut proteins was investigated. The optimized assay was relatively sensitive for the roasted peanut proteins. However, the binding ability of chicken IgYs to peanut proteins was found to be significantly altered by denaturation and hydrolysis of proteins. It was also observed that the effect of Millard chemistry on the detectability of peanut protein was less pronounced at high temperatures than at low temperatures. Copyright © 2013 Elsevier Ltd. All rights reserved.

Analysis of electric moments of RNA-binding proteins: implications for mechanism and prediction

Directory of Open Access Journals (Sweden)

Sarai Akinori

2011-02-01

Full Text Available Abstract Background Protein-RNA interactions play important role in many biological processes such as gene regulation, replication, protein synthesis and virus assembly. Although many structures of various types of protein-RNA complexes have been determined, the mechanism of protein-RNA recognition remains elusive. We have earlier shown that the simplest electrostatic properties viz. charge, dipole and quadrupole moments, calculated from backbone atomic coordinates of proteins are biased relative to other proteins, and these quantities can be used to identify DNA-binding proteins. Closely related, RNA-binding proteins are investigated in this study. In particular, discrimination between various types of RNA-binding proteins, evolutionary conservation of these bulk electrostatic features and effect of conformational changes by complex formation are investigated. Basic binding mechanism of a putative RNA-binding protein (HI1333 from Haemophilus influenza is suggested as a potential application of this study. Results We found that similar to DNA-binding proteins (DBPs, RNA-binding proteins (RBPs also show significantly higher values of electric moments. However, higher moments in RBPs are found to strongly depend on their functional class: proteins binding to ribosomal RNA (rRNA constitute the only class with all three of the properties (charge, dipole and quadrupole moments being higher than control proteins. Neural networks were trained using leave-one-out cross-validation to predict RBPs from control data as well as pair-wise classification capacity between proteins binding to various RNA types. RBPs and control proteins reached up to 78% accuracy measured by the area under the ROC curve. Proteins binding to rRNA are found to be best distinguished (AUC = 79%. Changes in dipole and quadrupole moments between unbound and bound structures were small and these properties are found to be robust under complex formation. Conclusions Bulk electric

ZnO nanorod biosensor for highly sensitive detection of specific protein binding

International Nuclear Information System (INIS)

Kim, Jin Suk; Park, Won Il; Lee, Chul Ho; Yi, Gyu Chul

2006-01-01

We report on the fabrication of electrical biosensors based on functionalized ZnO nanorod surfaces with biotin for highly sensitive detection of biological molecules. Due to the clean interface and easy surface modification, the ZnO nanorod sensors can easily detect streptavidin binding down to a concentration of 25 nM, which is more sensitive than previously reported one-dimensional (1D) nanostructure electrical biosensors. In addition, the unique device structure with a micrometer-scale hole at the center of the ZnO nanorod's conducting channel reduces the leakage current from the aqueous solution, hence enhancing device sensitivity. Moreover, ZnO nanorod field-effect-transistor (FET) sensors may open up opportunities to create many other oxide nanorod electrical sensors for highly sensitive and selective real-time detection of a wide variety of biomolecules.

CMsearch: simultaneous exploration of protein sequence space and structure space improves not only protein homology detection but also protein structure prediction

KAUST Repository

Cui, Xuefeng; Lu, Zhiwu; Wang, Sheng; Jing-Yan Wang, Jim; Gao, Xin

2016-01-01

Motivation: Protein homology detection, a fundamental problem in computational biology, is an indispensable step toward predicting protein structures and understanding protein functions. Despite the advances in recent decades on sequence alignment

Detecting significant changes in protein abundance

Directory of Open Access Journals (Sweden)

Kai Kammers

2015-06-01

Full Text Available We review and demonstrate how an empirical Bayes method, shrinking a protein's sample variance towards a pooled estimate, leads to far more powerful and stable inference to detect significant changes in protein abundance compared to ordinary t-tests. Using examples from isobaric mass labelled proteomic experiments we show how to analyze data from multiple experiments simultaneously, and discuss the effects of missing data on the inference. We also present easy to use open source software for normalization of mass spectrometry data and inference based on moderated test statistics.

Carbon nanotube-based sensing devices for human Arginase-1 detection

Directory of Open Access Journals (Sweden)

S. Baldo

2016-03-01

Full Text Available A new carbon nanotube-based device for detection of Arginase 1 (ARG-1 was produced. Multi-walled carbon nanotubes (MWCNTs were deposited between electrodes by dielectrophoresis (DEP in an accurate and reproducible way. This deposition method has the advantages of low cost and room temperature conditions and therefore, can be used on different kinds of substrates (silicon, glass, plastics allowing for large scale production of chemical or biological sensors. Scanning electrical microscope (SEM and electrical characterization have been performed on the biosensors before and after protein exposure. The devices were tested in the present work for the detection of ARG-1. They show high sensitivity and reproducibility, and can be easily and suitably modified to detect other proteins. Keywords: Carbon nanotube, Biosensor, Arginase, Dielectrophoresis, Biomarker, Protein

Isoelectric Point, Electric Charge, and Nomenclature of the Acid-Base Residues of Proteins

Science.gov (United States)

Maldonado, Andres A.; Ribeiro, Joao M.; Sillero, Antonio

2010-01-01

The main object of this work is to present the pedagogical usefulness of the theoretical methods, developed in this laboratory, for the determination of the isoelectric point (pI) and the net electric charge of proteins together with some comments on the naming of the acid-base residues of proteins. (Contains 8 figures and 4 tables.)

Silicon nanowire field-effect transistors for the detection of proteins

Science.gov (United States)

Madler, Carsten

In this dissertation I present results on our efforts to increase the sensitivity and selectivity of silicon nanowire ion-sensitive field-effect transistors for the detection of biomarkers, as well as a novel method for wireless power transfer based on metamaterial rectennas for their potential use as implantable sensors. The sensing scheme is based on changes in the conductance of the semiconducting nanowires upon binding of charged entities to the surface, which induces a field-effect. Monitoring the differential conductance thus provides information of the selective binding of biological molecules of interest to previously covalently linked counterparts on the nanowire surface. In order to improve on the performance of the nanowire sensing, we devised and fabricated a nanowire Wheatstone bridge, which allows canceling out of signal drift due to thermal fluctuations and dynamics of fluid flow. We showed that balancing the bridge significantly improves the signal-to-noise ratio. Further, we demonstrated the sensing of novel melanoma biomarker TROY at clinically relevant concentrations and distinguished it from nonspecific binding by comparing the reaction kinetics. For increased sensitivity, an amplification method was employed using an enzyme which catalyzes a signal-generating reaction by changing the redox potential of a redox pair. In addition, we investigated the electric double layer, which forms around charges in an electrolytic solution. It causes electrostatic screening of the proteins of interest, which puts a fundamental limitation on the biomarker detection in solutions with high salt concentrations, such as blood. We solved the coupled Nernst-Planck and Poisson equations for the electrolyte under influence of an oscillating electric field and discovered oscillations of the counterion concentration at a characteristic frequency. In addition to exploring different methods for improved sensing capabilities, we studied an innovative method to supply power

Microscopic Void Detection for Predicting Remaining Life in Electric Cable Insulation

International Nuclear Information System (INIS)

Horvath, David A.; Avila, Steven M.

2003-01-01

A reliable method of testing for remaining life in electric cable insulation has continued to elude the nuclear industry as it seeks to extend the life and license of its nuclear stations. Until recently, a trendable, measurable electrical property has not been found, and unexpected cable failures continue to be reported. Most reliable approaches to date rely on monitoring mechanical properties, which are assumed to degrade faster than the insulation's electrical properties. This paper introduces a promising technique based on void characterization, which is dependent on an electrical property related to dielectric strength. A relationship between insulation void characteristics (size and density) and the onset of partial discharge is known to exist. A similar relationship can be shown between void characteristics and unacceptable leakage currents (another typical cable failure criterion). For low-voltage cables, it is believed void content can be correlated to mechanical property degradation.This paper will report on an approach for using void information, research results showing the existence of trendable void characteristics in commonly used electric insulation materials, and techniques for detecting the voids (both laboratory- and field-based techniques). Acoustical microscopy was found to be potentially more suitable than conventional ultrasound for nondestructive in situ detection and monitoring of void characteristics in jacketed multiconductor insulation while ignoring the jacket. Also, optical and scanning electron microscope techniques will play an essential role in establishing the database necessary for continued development and implementation of this promising technique

A storage ring experiment to detect a proton electric dipole moment

Energy Technology Data Exchange (ETDEWEB)

Anastassopoulos, V. [Department of Physics, University of Patras, 26500 Rio-Patras, Greece; Andrianov, S. [Faculty of Applied Mathematics and Control Processes, Saint-Petersburg State University, Saint-Petersburg, Russia; Baartman, R. [TRIUMF, 4004 Wesbrook Mall, Vancouver, British Columbia V6T2A3, Canada; Baessler, S. [Department of Physics, University of Virginia, Charlottesville, Virginia 22904, USA; Bai, M. [Institut für Kernphysik and JARA-Fame, Forschungszentrum Jülich, 52425 Jülich, Germany; Benante, J. [Brookhaven National Laboratory, Upton, New York 11973, USA; Berz, M. [Department of Physics and Astronomy, Michigan State University, East Lansing, Michigan 48824, USA; Blaskiewicz, M. [Brookhaven National Laboratory, Upton, New York 11973, USA; Bowcock, T. [Department of Physics, University of Liverpool, Liverpool, United Kingdom; Brown, K. [Brookhaven National Laboratory, Upton, New York 11973, USA; Casey, B. [Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA; Conte, M. [Physics Department and INFN Section of Genoa, 16146 Genoa, Italy; Crnkovic, J. D. [Brookhaven National Laboratory, Upton, New York 11973, USA; D’Imperio, N. [Brookhaven National Laboratory, Upton, New York 11973, USA; Fanourakis, G. [Institute of Nuclear and Particle Physics NCSR Demokritos, GR-15310 Aghia Paraskevi Athens, Greece; Fedotov, A. [Brookhaven National Laboratory, Upton, New York 11973, USA; Fierlinger, P. [Technical University München, Physikdepartment and Excellence-Cluster “Universe,” Garching, Germany; Fischer, W. [Brookhaven National Laboratory, Upton, New York 11973, USA; Gaisser, M. O. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Giomataris, Y. [CEA/Saclay, DAPNIA, 91191 Gif-sur-Yvette Cedex, France; Grosse-Perdekamp, M. [Department of Physics, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA; Guidoboni, G. [University of Ferrara, INFN of Ferrara, Ferrara, Italy; Hacıömeroğlu, S. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Hoffstaetter, G. [Laboratory for Elementary-Particle Physics, Cornell University, Ithaca, New York 14853, USA; Huang, H. [Brookhaven National Laboratory, Upton, New York 11973, USA; Incagli, M. [Physics Department, University and INFN Pisa, Pisa, Italy; Ivanov, A. [Faculty of Applied Mathematics and Control Processes, Saint-Petersburg State University, Saint-Petersburg, Russia; Kawall, D. [Department of Physics, University of Massachusetts, Amherst, Massachusetts 01003, USA; Kim, Y. I. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; King, B. [Department of Physics, University of Liverpool, Liverpool, United Kingdom; Koop, I. A. [Budker Institute of Nuclear Physics, 630090 Novosibirsk, Russia; Lazarus, D. M. [Brookhaven National Laboratory, Upton, New York 11973, USA; Lebedev, V. [Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA; Lee, M. J. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Lee, S. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Lee, Y. H. [Korea Research Institute of Standards and Science, Daejeon 34141, South Korea; Lehrach, A. [Institut für Kernphysik and JARA-Fame, Forschungszentrum Jülich, 52425 Jülich, Germany; RWTH Aachen University and JARA-Fame, III. Physikalisches Institut B, Physikzentrum, 52056 Aachen, Germany; Lenisa, P. [University of Ferrara, INFN of Ferrara, Ferrara, Italy; Levi Sandri, P. [Laboratori Nazionali di Frascati, INFN, I-00044 Frascati, Rome, Italy; Luccio, A. U. [Brookhaven National Laboratory, Upton, New York 11973, USA; Lyapin, A. [Royal Holloway, University of London, Egham, Surrey, United Kingdom; MacKay, W. [Brookhaven National Laboratory, Upton, New York 11973, USA; Maier, R. [Institut für Kernphysik and JARA-Fame, Forschungszentrum Jülich, 52425 Jülich, Germany; Makino, K. [Department of Physics and Astronomy, Michigan State University, East Lansing, Michigan 48824, USA; Malitsky, N. [Brookhaven National Laboratory, Upton, New York 11973, USA; Marciano, W. J. [Brookhaven National Laboratory, Upton, New York 11973, USA; Meng, W. [Brookhaven National Laboratory, Upton, New York 11973, USA; Meot, F. [Brookhaven National Laboratory, Upton, New York 11973, USA; Metodiev, E. M. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Harvard College, Harvard University, Cambridge, Massachusetts 02138, USA; Miceli, L. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Moricciani, D. [Dipartimento di Fisica dell’Univ. di Roma “Tor Vergata” and INFN Sezione di Roma Tor Vergata, Rome, Italy; Morse, W. M. [Brookhaven National Laboratory, Upton, New York 11973, USA; Nagaitsev, S. [Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA; Nayak, S. K. [Brookhaven National Laboratory, Upton, New York 11973, USA; Orlov, Y. F. [Laboratory for Elementary-Particle Physics, Cornell University, Ithaca, New York 14853, USA; Ozben, C. S. [Istanbul Technical University, Istanbul 34469, Turkey; Park, S. T. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Pesce, A. [University of Ferrara, INFN of Ferrara, Ferrara, Italy; Petrakou, E. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Pile, P. [Brookhaven National Laboratory, Upton, New York 11973, USA; Podobedov, B. [Brookhaven National Laboratory, Upton, New York 11973, USA; Polychronakos, V. [Brookhaven National Laboratory, Upton, New York 11973, USA; Pretz, J. [RWTH Aachen University and JARA-Fame, III. Physikalisches Institut B, Physikzentrum, 52056 Aachen, Germany; Ptitsyn, V. [Brookhaven National Laboratory, Upton, New York 11973, USA; Ramberg, E. [Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA; Raparia, D. [Brookhaven National Laboratory, Upton, New York 11973, USA; Rathmann, F. [Institut für Kernphysik and JARA-Fame, Forschungszentrum Jülich, 52425 Jülich, Germany; Rescia, S. [Brookhaven National Laboratory, Upton, New York 11973, USA; Roser, T. [Brookhaven National Laboratory, Upton, New York 11973, USA; Kamal Sayed, H. [Brookhaven National Laboratory, Upton, New York 11973, USA; Semertzidis, Y. K. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Department of Physics, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, South Korea; Senichev, Y. [Institut für Kernphysik and JARA-Fame, Forschungszentrum Jülich, 52425 Jülich, Germany; Sidorin, A. [Joint Institute for Nuclear Research, Dubna, Moscow region, Russia; Silenko, A. [Joint Institute for Nuclear Research, Dubna, Moscow region, Russia; Research Institute for Nuclear Problems of Belarusian State University, Minsk, Belarus; Simos, N. [Brookhaven National Laboratory, Upton, New York 11973, USA; Stahl, A. [RWTH Aachen University and JARA-Fame, III. Physikalisches Institut B, Physikzentrum, 52056 Aachen, Germany; Stephenson, E. J. [Indiana University Center for Spacetime Symmetries, Bloomington, Indiana 47405, USA; Ströher, H. [Institut für Kernphysik and JARA-Fame, Forschungszentrum Jülich, 52425 Jülich, Germany; Syphers, M. J. [Fermi National Accelerator Laboratory, Batavia, Illinois 60510, USA; Department of Physics, Northern Illinois University, DeKalb, Illinois 60115, USA; Talman, J. [Brookhaven National Laboratory, Upton, New York 11973, USA; Talman, R. M. [Laboratory for Elementary-Particle Physics, Cornell University, Ithaca, New York 14853, USA; Tishchenko, V. [Brookhaven National Laboratory, Upton, New York 11973, USA; Touramanis, C. [Department of Physics, University of Liverpool, Liverpool, United Kingdom; Tsoupas, N. [Brookhaven National Laboratory, Upton, New York 11973, USA; Venanzoni, G. [Laboratori Nazionali di Frascati, INFN, I-00044 Frascati, Rome, Italy; Vetter, K. [Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831, USA; Vlassis, S. [Department of Physics, University of Patras, 26500 Rio-Patras, Greece; Won, E. [Center for Axion and Precision Physics Research, Institute for Basic Science (IBS), Daejeon 34141, South Korea; Physics Department, Korea University, Seoul 02841, South Korea; Zavattini, G. [University of Ferrara, INFN of Ferrara, Ferrara, Italy; Zelenski, A. [Brookhaven National Laboratory, Upton, New York 11973, USA; Zioutas, K. [Department of Physics, University of Patras, 26500 Rio-Patras, Greece

2016-11-01

A new experiment is described to detect a permanent electric dipole moment of the proton with a sensitivity of $10^{-29}e\\cdot$cm by using polarized "magic" momentum $0.7$~GeV/c protons in an all-electric storage ring. Systematic errors relevant to the experiment are discussed and techniques to address them are presented. The measurement is sensitive to new physics beyond the Standard Model at the scale of 3000~TeV.

A storage ring experiment to detect a proton electric dipole moment.

Science.gov (United States)

Anastassopoulos, V; Andrianov, S; Baartman, R; Baessler, S; Bai, M; Benante, J; Berz, M; Blaskiewicz, M; Bowcock, T; Brown, K; Casey, B; Conte, M; Crnkovic, J D; D'Imperio, N; Fanourakis, G; Fedotov, A; Fierlinger, P; Fischer, W; Gaisser, M O; Giomataris, Y; Grosse-Perdekamp, M; Guidoboni, G; Hacıömeroğlu, S; Hoffstaetter, G; Huang, H; Incagli, M; Ivanov, A; Kawall, D; Kim, Y I; King, B; Koop, I A; Lazarus, D M; Lebedev, V; Lee, M J; Lee, S; Lee, Y H; Lehrach, A; Lenisa, P; Levi Sandri, P; Luccio, A U; Lyapin, A; MacKay, W; Maier, R; Makino, K; Malitsky, N; Marciano, W J; Meng, W; Meot, F; Metodiev, E M; Miceli, L; Moricciani, D; Morse, W M; Nagaitsev, S; Nayak, S K; Orlov, Y F; Ozben, C S; Park, S T; Pesce, A; Petrakou, E; Pile, P; Podobedov, B; Polychronakos, V; Pretz, J; Ptitsyn, V; Ramberg, E; Raparia, D; Rathmann, F; Rescia, S; Roser, T; Kamal Sayed, H; Semertzidis, Y K; Senichev, Y; Sidorin, A; Silenko, A; Simos, N; Stahl, A; Stephenson, E J; Ströher, H; Syphers, M J; Talman, J; Talman, R M; Tishchenko, V; Touramanis, C; Tsoupas, N; Venanzoni, G; Vetter, K; Vlassis, S; Won, E; Zavattini, G; Zelenski, A; Zioutas, K

2016-11-01

A new experiment is described to detect a permanent electric dipole moment of the proton with a sensitivity of 10 -29 e ⋅ cm by using polarized "magic" momentum 0.7 GeV/c protons in an all-electric storage ring. Systematic errors relevant to the experiment are discussed and techniques to address them are presented. The measurement is sensitive to new physics beyond the standard model at the scale of 3000 TeV.

High resolution in-operando microimaging of solar cells with pulsed electrically-detected magnetic resonance

Science.gov (United States)

Katz, Itai; Fehr, Matthias; Schnegg, Alexander; Lips, Klaus; Blank, Aharon

2015-02-01

The in-operando detection and high resolution spatial imaging of paramagnetic defects, impurities, and states becomes increasingly important for understanding loss mechanisms in solid-state electronic devices. Electron spin resonance (ESR), commonly employed for observing these species, cannot meet this challenge since it suffers from limited sensitivity and spatial resolution. An alternative and much more sensitive method, called electrically-detected magnetic resonance (EDMR), detects the species through their magnetic fingerprint, which can be traced in the device's electrical current. However, until now it could not obtain high resolution images in operating electronic devices. In this work, the first spatially-resolved electrically-detected magnetic resonance images (EDMRI) of paramagnetic states in an operating real-world electronic device are provided. The presented method is based on a novel microwave pulse sequence allowing for the coherent electrical detection of spin echoes in combination with powerful pulsed magnetic-field gradients. The applicability of the method is demonstrated on a device-grade 1-μm-thick amorphous silicon (a-Si:H) solar cell and an identical device that was degraded locally by an electron beam. The degraded areas with increased concentrations of paramagnetic defects lead to a local increase in recombination that is mapped by EDMRI with ∼20-μm-scale pixel resolution. The novel approach presented here can be widely used in the nondestructive in-operando three-dimensional characterization of solid-state electronic devices with a resolution potential of less than 100 nm.

Miniaturized Aptamer-Based Assays for Protein Detection

Directory of Open Access Journals (Sweden)

Alessandro Bosco

2016-09-01

Full Text Available The availability of devices for cancer biomarker detection at early stages of the disease is one of the most critical issues in biomedicine. Towards this goal, to increase the assay sensitivity, device miniaturization strategies empowered by the employment of high affinity protein binders constitute a valuable approach. In this work we propose two different surface-based miniaturized platforms for biomarker detection in body fluids: the first platform is an atomic force microscopy (AFM-based nanoarray, where AFM is used to generate functional nanoscale areas and to detect biorecognition through careful topographic measurements; the second platform consists of a miniaturized electrochemical cell to detect biomarkers through electrochemical impedance spectroscopy (EIS analysis. Both devices rely on robust and highly-specific protein binders as aptamers, and were tested for thrombin detection. An active layer of DNA-aptamer conjugates was immobilized via DNA directed immobilization on complementary single-stranded DNA self-assembled monolayers confined on a nano/micro area of a gold surface. Results obtained with these devices were compared with the output of surface plasmon resonance (SPR assays used as reference. We succeeded in capturing antigens in concentrations as low as a few nM. We put forward ideas to push the sensitivity further to the pM range, assuring low biosample volume (μL range assay conditions.

Detection of questionable occlusal carious lesions using an electrical bioimpedance method with fractional electrical model

Energy Technology Data Exchange (ETDEWEB)

Morais, A. P. [Biomedical Engineering Program, COPPE, Rio de Janeiro (Brazil); Salgado de Oliveira University, Marechal Deodoro Street, 217 – Centro, Niterói, Rio de Janeiro (Brazil); Pino, A. V. [Biomedical Engineering Program, COPPE, Rio de Janeiro (Brazil); Souza, M. N. [Biomedical Engineering Program, COPPE, Rio de Janeiro (Brazil); Electronics Department at Polytechnic School, Federal University of Rio de Janeiro, Centro de Tecnologia Bloco H sala 217, Ilha do Fundão, Rio de Janeiro (Brazil)

2016-08-15

This in vitro study evaluated the diagnostic performance of an alternative electric bioimpedance spectroscopy technique (BIS-STEP) detect questionable occlusal carious lesions. Six specialists carried out the visual (V), radiography (R), and combined (VR) exams of 57 sound or non-cavitated occlusal carious lesion teeth classifying the occlusal surfaces in sound surface (H), enamel caries (EC), and dentinal caries (DC). Measurements were based on the current response to a step voltage excitation (BIS-STEP). A fractional electrical model was used to predict the current response in the time domain and to estimate the model parameters: Rs and Rp (resistive parameters), and C and α (fractional parameters). Histological analysis showed caries prevalence of 33.3% being 15.8% hidden caries. Combined examination obtained the best traditional diagnostic results with specificity = 59.0%, sensitivity = 70.9%, and accuracy = 60.8%. There were statistically significant differences in bioimpedance parameters between the H and EC groups (p = 0.016) and between the H and DC groups (Rs, p = 0.006; Rp, p = 0.022, and α, p = 0.041). Using a suitable threshold for the Rs, we obtained specificity = 60.7%, sensitivity = 77.9%, accuracy = 73.2%, and 100% of detection for deep lesions. It can be concluded that BIS-STEP method could be an important tool to improve the detection and management of occlusal non-cavitated primary caries and pigmented sites.

Detection of questionable occlusal carious lesions using an electrical bioimpedance method with fractional electrical model

International Nuclear Information System (INIS)

Morais, A. P.; Pino, A. V.; Souza, M. N.

2016-01-01

This in vitro study evaluated the diagnostic performance of an alternative electric bioimpedance spectroscopy technique (BIS-STEP) detect questionable occlusal carious lesions. Six specialists carried out the visual (V), radiography (R), and combined (VR) exams of 57 sound or non-cavitated occlusal carious lesion teeth classifying the occlusal surfaces in sound surface (H), enamel caries (EC), and dentinal caries (DC). Measurements were based on the current response to a step voltage excitation (BIS-STEP). A fractional electrical model was used to predict the current response in the time domain and to estimate the model parameters: Rs and Rp (resistive parameters), and C and α (fractional parameters). Histological analysis showed caries prevalence of 33.3% being 15.8% hidden caries. Combined examination obtained the best traditional diagnostic results with specificity = 59.0%, sensitivity = 70.9%, and accuracy = 60.8%. There were statistically significant differences in bioimpedance parameters between the H and EC groups (p = 0.016) and between the H and DC groups (Rs, p = 0.006; Rp, p = 0.022, and α, p = 0.041). Using a suitable threshold for the Rs, we obtained specificity = 60.7%, sensitivity = 77.9%, accuracy = 73.2%, and 100% of detection for deep lesions. It can be concluded that BIS-STEP method could be an important tool to improve the detection and management of occlusal non-cavitated primary caries and pigmented sites.

Security attack detection algorithm for electric power gis system based on mobile application

Science.gov (United States)

Zhou, Chao; Feng, Renjun; Wang, Liming; Huang, Wei; Guo, Yajuan

2017-05-01

Electric power GIS is one of the key information technologies to satisfy the power grid construction in China, and widely used in power grid construction planning, weather, and power distribution management. The introduction of electric power GIS based on mobile applications is an effective extension of the geographic information system that has been widely used in the electric power industry. It provides reliable, cheap and sustainable power service for the country. The accurate state estimation is the important conditions to maintain the normal operation of the electric power GIS. Recent research has shown that attackers can inject the complex false data into the power system. The injection attack of this new type of false data (load integrity attack LIA) can successfully bypass the routine detection to achieve the purpose of attack, so that the control center will make a series of wrong decision. Eventually, leading to uneven distribution of power in the grid. In order to ensure the safety of the electric power GIS system based on mobile application, it is very important to analyze the attack mechanism and propose a new type of attack, and to study the corresponding detection method and prevention strategy in the environment of electric power GIS system based on mobile application.

Electrochemical Aptamer Scaffold Biosensors for Detection of Botulism and Ricin Proteins.

Science.gov (United States)

Daniel, Jessica; Fetter, Lisa; Jett, Susan; Rowland, Teisha J; Bonham, Andrew J

2017-01-01

Electrochemical DNA (E-DNA) biosensors enable the detection and quantification of a variety of molecular targets, including oligonucleotides, small molecules, heavy metals, antibodies, and proteins. Here we describe the design, electrode preparation and sensor attachment, and voltammetry conditions needed to generate and perform measurements using E-DNA biosensors against two protein targets, the biological toxins ricin and botulinum neurotoxin. This method can be applied to generate E-DNA biosensors for the detection of many other protein targets, with potential advantages over other systems including sensitive detection limits typically in the nanomolar range, real-time monitoring, and reusable biosensors.

Effects on DPPH inhibition of egg-white protein polypeptides treated by pulsed electric field technology.

Science.gov (United States)

Wang, Ke; Wang, Jia; Liu, Bolong; Lin, Songyi; Zhao, Ping; Liu, Jingbo; Jones, Gregory; Huang, Hsiang-Chi

2013-05-01

Egg-white protein polypeptides are potentially used as a functional ingredient in food products. In this study, the effects on DPPH inhibition of egg-white protein polypeptides ranging from 10 to 30 kDa treated by pulsed electric field (PEF) technology were investigated. 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) inhibition (%) was used to evaluate the antioxidant activity of polypeptides. In order to develop and optimize a pulsed electric field (PEF) mathematical model for improving the antioxidant activity, we have investigated three variables, including concentration (6, 8 and 10 mg mL(-1)), electric field intensity (10, 20 and 30 kV cm(-1)) and pulse frequency (2000, 2350 and 2700 Hz) and subsequently optimized them by response surface methodology (RSM). The concentration (8 mg mL(-1)), electric field intensity (10 kV cm(-1)) and pulse frequency (2000 Hz) were found to be the optimal conditions under which the DPPH inhibition increased 28.44%, compared to the sample without PEF treatment. Both near-infrared spectroscopy (NIR) and mid-infrared spectroscopy (MIR) were used to analyze the change of functional groups. The results showed that PEF technology could improve the antioxidant activity of antioxidant polypeptides from egg-white protein under the optimized conditions. © 2012 Society of Chemical Industry.

Direct protein detection with a nano-interdigitated array gate MOSFET.

Science.gov (United States)

Tang, Xiaohui; Jonas, Alain M; Nysten, Bernard; Demoustier-Champagne, Sophie; Blondeau, Franoise; Prévot, Pierre-Paul; Pampin, Rémi; Godfroid, Edmond; Iñiguez, Benjamin; Colinge, Jean-Pierre; Raskin, Jean-Pierre; Flandre, Denis; Bayot, Vincent

2009-08-15

A new protein sensor is demonstrated by replacing the gate of a metal oxide semiconductor field effect transistor (MOSFET) with a nano-interdigitated array (nIDA). The sensor is able to detect the binding reaction of a typical antibody Ixodes ricinus immunosuppressor (anti-Iris) protein at a concentration lower than 1 ng/ml. The sensor exhibits a high selectivity and reproducible specific detection. We provide a simple model that describes the behavior of the sensor and explains the origin of its high sensitivity. The simulated and experimental results indicate that the drain current of nIDA-gate MOSFET sensor is significantly increased with the successive binding of the thiol layer, Iris and anti-Iris protein layers. It is found that the sensor detection limit can be improved by well optimizing the geometrical parameters of nIDA-gate MOSFET. This nanobiosensor, with real-time and label-free capabilities, can easily be used for the detection of other proteins, DNA, virus and cancer markers. Moreover, an on-chip associated electronics nearby the sensor can be integrated since its fabrication is compatible with complementary metal oxide semiconductor (CMOS) technology.

Detection of protein kinase activity by renaturation in sodium dodecyl sulfate-polyacrylamide gels

International Nuclear Information System (INIS)

Anostario, M. Jr.; Harrison, M.L.; Geahlen, R.L.

1986-01-01

The authors have developed a procedure for identifying protein kinase activity in protein samples following electrophoresis on SDS-polyacrylamide gels. Proteins are allowed to renature directly in the gel by removal of detergent. The gel is then incubated with [γ- 32 P]ATP to allow renatured protein kinases to autophosphorylate or to phosphorylate various substrates which can be incorporated into the gel. The positions of the radiolabeled proteins can then be detected by autoradiography. With this technique, using purified catalytic subunit of cAMP-dependent protein kinase, enzyme concentrations as low as 0.01 μg can be detected on gels containing 1.0 mg/ml casein. The procedure is also applicable for the determination of active subunits of multisubunit protein kinases. For example, when the two subunits of casein kinase II are separated by SDS-polyacrylamide gel electrophoresis and allowed to renature, only the larger α subunit shows activity. This procedure can also be used to detect and distinguish kinases present in heterogeneous mixtures. Starting with a particulate fraction from LSTRA, a murine T cell lymphoma, several distinct enzymes were detected, including a 30,000 Dalton protein with protein-tyrosine kinase activity. This same enzyme has also been detected in T lymphocytes and other T lymphoid cell lines

Bumble Bees (Bombus terrestris use mechanosensory hairs to detect electric fields

Directory of Open Access Journals (Sweden)

Sutton Gregory

2016-01-01

Full Text Available Bees and flowers have an intricate relationship which benefits both organisms. Plants provide nectar bees, in turn, distribute pollen to fertilize plants. To make pollination work, flowers need a mechanism to incentivize individual bees to visit only a single species of flower. Flowers, like modern advertising agencies, use multiple senses to create a floral ‘brand’ that is easily recognized. Size, smell, colour, touch, and even temperature are used to allow bees to differentiate between flower species. Recently, a new sense has been found that is usable by bees to differentiate flowers, an ‘electric sense’: they can identify flowers based only on the flower’s electric field. This new sense provides a novel example of how flowers differentiate themselves to bees and has obvious implications for how bees and flowers interact with the electrical world around us. Bumble bees detect this electric field by using their body hairs, which bend in the presence of electric charge.

An electronic channel switching-based aptasensor for ultrasensitive protein detection

Energy Technology Data Exchange (ETDEWEB)

Li Hongbo; Wang Cui [State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082 (China); Wu Zaisheng, E-mail: wuzaisheng@163.com [State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082 (China); Lu Limin; Qiu Liping; Zhou Hui; Shen Guoli [State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082 (China); Yu Ruqin, E-mail: rqyu@hnu.edu.cn [State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082 (China)

2013-01-03

Highlights: Black-Right-Pointing-Pointer Target IgE is successfully designed to serve as a barrier to separate enzyme from its substrate. Black-Right-Pointing-Pointer This sensing platform of electronic channel switching-based aptasensor can be simply manipulated. Black-Right-Pointing-Pointer The stable hairpin structure of anti-IgE aptamer is utilized to detect target IgE. Black-Right-Pointing-Pointer The sensor is ultrasensitive sensitivity, excellent selectivity and small volume of sample. Black-Right-Pointing-Pointer It is a powerful platform to be further expanded to detect more kinds of proteins and even cells. - Abstract: Due to the ubiquity and essential of the proteins in all living organisms, the identification and quantification of disease-specific proteins are particularly important. Because the conformational change of aptamer upon its target or probe/target/probe sandwich often is the primary prerequisite for the design of an electrochemical aptameric assay system, it is extremely difficult to construct the electrochemical aptasensor for protein assay because the corresponding aptamers cannot often meet the requirement. To circumvent the obstacles mentioned, an electronic channel switching-based (ECS) aptasensor for ultrasensitive protein detection is developed. The essential achievement made is that an innovative sensing concept is proposed: the hairpin structure of aptamer is designed to pull electroactive species toward electrode surface and makes the surface-immobilized IgE serve as a barrier that separates enzyme from its substrate. It seemingly ensures that the ECS aptasensor exhibits most excellent assay features, such as, a detection limit of 4.44 Multiplication-Sign 10{sup -6} {mu}g mL{sup -1} (22.7 fM, 220 zmol in 10-{mu}L sample) (demonstrating a 5 orders of magnitude improvement in detection sensitivity compared with classical electronic aptasensors) and dynamic response range from 4.44 Multiplication-Sign 10{sup -6} to 4.44 Multiplication

Modified electrical survey for effective leakage detection at concrete hydraulic facilities

Science.gov (United States)

Lee, Bomi; Oh, Seokhoon

2018-02-01

Three original electrode arrays for the effective leakage detection of concrete hydraulic facilities through electrical resistivity surveys are proposed: 'cross-potential', 'direct-potential' and modified tomography-like arrays. The main differences with respect to the commonly used arrays are that the current line-sources are separated from potential pole lines and floated upon the water. The potential pole lines are located directly next to the facility in order to obtain intuitive data and useful interpretations of the internal conditions of the hydraulic facility. This modified configuration of the array clearly displays the horizontal variation of the electrical field around the damaged zones of the concrete hydraulic facility, and any anomalous regions that might be found between potential poles placed across the facilities. In order to facilitate the interpretation of these modified electrical surveys, a new and creative way of presenting the measurements is also proposed and an inversion approach is provided for the modified tomography-like array. A numerical modeling and two field tests were performed to verify these new arrays and interpretation methods. The cross and direct potential array implied an ability to detect small variations of the potential field near the measurement poles. The proposed array showed the overall potential distribution across the hydraulic facility which may be used to assist in the search of trouble zones within the structure, in combination with the traditional electrical resistivity array.

Comparative sensitivity of 125I-protein A and enzyme-conjugated antibodies for detection of immunoblotted proteins

International Nuclear Information System (INIS)

Bernstein, J.M.; Stokes, C.E.; Fernie, B.

1987-01-01

Immunoblotting is a powerful technique for the detection of small amounts of immunologically interesting proteins in unpurified preparations. Iodinated protein A (PA) has been widely used as a second antibody for detection of proteins; however, it does not bind equally well to immunoglobulins from different species nor does it bind to all subclasses of immunoglobulin G (IgG). We compared the sensitivity of [ 125 I]PA with those of both horseradish peroxidase-conjugated second antibodies (HRP) and glucose oxidase-anti-glucose oxidase (GAG) soluble complexes for visualizing bovine serum albumin, human IgG, or human C3 which was either dot blotted or electroblotted to nitrocellulose. [ 125 I]PA was uniformly 10- to 100-fold less sensitive than either HRP or GAG. GAG was more sensitive than HRP except for C3 (electroblotting) and bovine serum albumin and IgG (dot blotting), in which they were equivalent. In general, dot blotting was 10- to 1000-fold more sensitive than electroblotting. Although relative sensitivities varied depending on the proteins analyzed and the antisera used, GAG appeared to be superior to [ 125 I]PA and HRP for detection of immunoblotted proteins

HPTLC-aptastaining - Innovative protein detection system for high-performance thin-layer chromatography

Science.gov (United States)

Morschheuser, Lena; Wessels, Hauke; Pille, Christina; Fischer, Judith; Hünniger, Tim; Fischer, Markus; Paschke-Kratzin, Angelika; Rohn, Sascha

2016-05-01

Protein analysis using high-performance thin-layer chromatography (HPTLC) is not commonly used but can complement traditional electrophoretic and mass spectrometric approaches in a unique way. Due to various detection protocols and possibilities for hyphenation, HPTLC protein analysis is a promising alternative for e.g., investigating posttranslational modifications. This study exemplarily focused on the investigation of lysozyme, an enzyme which is occurring in eggs and technologically added to foods and beverages such as wine. The detection of lysozyme is mandatory, as it might trigger allergenic reactions in sensitive individuals. To underline the advantages of HPTLC in protein analysis, the development of innovative, highly specific staining protocols leads to improved sensitivity for protein detection on HPTLC plates in comparison to universal protein derivatization reagents. This study aimed at developing a detection methodology for HPTLC separated proteins using aptamers. Due to their affinity and specificity towards a wide range of targets, an aptamer based staining procedure on HPTLC (HPTLC-aptastaining) will enable manifold analytical possibilities. Besides the proof of its applicability for the very first time, (i) aptamer-based staining of proteins is applicable on different stationary phase materials and (ii) furthermore, it can be used as an approach for a semi-quantitative estimation of protein concentrations.

A Type-2 fuzzy data fusion approach for building reliable weighted protein interaction networks with application in protein complex detection.

Science.gov (United States)

Mehranfar, Adele; Ghadiri, Nasser; Kouhsar, Morteza; Golshani, Ashkan

2017-09-01

Detecting the protein complexes is an important task in analyzing the protein interaction networks. Although many algorithms predict protein complexes in different ways, surveys on the interaction networks indicate that about 50% of detected interactions are false positives. Consequently, the accuracy of existing methods needs to be improved. In this paper we propose a novel algorithm to detect the protein complexes in 'noisy' protein interaction data. First, we integrate several biological data sources to determine the reliability of each interaction and determine more accurate weights for the interactions. A data fusion component is used for this step, based on the interval type-2 fuzzy voter that provides an efficient combination of the information sources. This fusion component detects the errors and diminishes their effect on the detection protein complexes. So in the first step, the reliability scores have been assigned for every interaction in the network. In the second step, we have proposed a general protein complex detection algorithm by exploiting and adopting the strong points of other algorithms and existing hypotheses regarding real complexes. Finally, the proposed method has been applied for the yeast interaction datasets for predicting the interactions. The results show that our framework has a better performance regarding precision and F-measure than the existing approaches. Copyright © 2017 Elsevier Ltd. All rights reserved.

Radiometric immunosorbent assay for the detection of anti-hormone-binding protein antibodies

International Nuclear Information System (INIS)

Pierce, E.A.; Dame, M.C.; DeLuca, H.F.

1986-01-01

A radiometric immunosorbent assay (RISA) for the detection of monoclonal antibodies to hormone-binding proteins has been developed. The assay involves incubating hybridoma supernatants in microtiter wells that have been coated with goat anti-mouse IgG antibodies. Any mouse IgG in the test supernatant is thus specifically retained in the wells. Radioactive ligand-binding protein complexes are then incubated in the wells. The presence of anti-binding protein antibodies in the supernatant is indicated by specific retention of radioactive ligand-binding protein complexes in the wells. Crude antigen preparations, such as tissue homogenates, can be used to detect antibodies. The assay is capable of detecting antibody at concentrations 20 ng/ml (approx. 100 pM IgG). The RISA has been used successfully to screen for monoclonal antibodies to the intracellular receptor for 1,25-dihydroxyvitamin D 3 and should be useful for the detection of antibodies to ligand-binding proteins in general

Latest Progress of Fault Detection and Localization in Complex Electrical Engineering

Science.gov (United States)

Zhao, Zheng; Wang, Can; Zhang, Yagang; Sun, Yi

2014-01-01

In the researches of complex electrical engineering, efficient fault detection and localization schemes are essential to quickly detect and locate faults so that appropriate and timely corrective mitigating and maintenance actions can be taken. In this paper, under the current measurement precision of PMU, we will put forward a new type of fault detection and localization technology based on fault factor feature extraction. Lots of simulating experiments indicate that, although there are disturbances of white Gaussian stochastic noise, based on fault factor feature extraction principal, the fault detection and localization results are still accurate and reliable, which also identifies that the fault detection and localization technology has strong anti-interference ability and great redundancy.

Analytically Sensitive Protein Detection in Microtiter Plates by Proximity Ligation with Rolling Circle Amplification.

Science.gov (United States)

Ebai, Tonge; Souza de Oliveira, Felipe Marques; Löf, Liza; Wik, Lotta; Schweiger, Caroline; Larsson, Anders; Keilholtz, Ulrich; Haybaeck, Johannes; Landegren, Ulf; Kamali-Moghaddam, Masood

2017-09-01

Detecting proteins at low concentrations in plasma is crucial for early diagnosis. Current techniques in clinical routine, such as sandwich ELISA, provide sensitive protein detection because of a dependence on target recognition by pairs of antibodies, but detection of still lower protein concentrations is often called for. Proximity ligation assay with rolling circle amplification (PLARCA) is a modified proximity ligation assay (PLA) for analytically specific and sensitive protein detection via binding of target proteins by 3 antibodies, and signal amplification via rolling circle amplification (RCA) in microtiter wells, easily adapted to instrumentation in use in hospitals. Proteins captured by immobilized antibodies were detected using a pair of oligonucleotide-conjugated antibodies. Upon target recognition these PLA probes guided oligonucleotide ligation, followed by amplification via RCA of circular DNA strands that formed in the reaction. The RCA products were detected by horseradish peroxidase-labeled oligonucleotides to generate colorimetric reaction products with readout in an absorbance microplate reader. We compared detection of interleukin (IL)-4, IL-6, IL-8, p53, and growth differentiation factor 15 (GDF-15) by PLARCA and conventional sandwich ELISA or immuno-RCA. PLARCA detected lower concentrations of proteins and exhibited a broader dynamic range compared to ELISA and iRCA using the same antibodies. IL-4 and IL-6 were detected in clinical samples at femtomolar concentrations, considerably lower than for ELISA. PLARCA offers detection of lower protein levels and increased dynamic ranges compared to ELISA. The PLARCA procedure may be adapted to routine instrumentation available in hospitals and research laboratories. © 2017 American Association for Clinical Chemistry.

Manufacturing and testing flexible microfluidic devices with optical and electrical detection mechanisms

OpenAIRE

Ivan, M.G.; Vivet, F.; Meinders, E.R.

2010-01-01

Flexible microfluidic devices made of poly(dimethylsiloxane) (PDMS) were manufactured by soft lithography, and tested in detection of ionic species using optical absorption spectroscopy and electrical measurements. PDMS was chosen due to its flexibility and ease of surface modification by exposure to plasma and UV treatment, its transparency in UV-Vis regions of the light spectrum, and biocompatibility. The dual-detection mechanism allows the user more freedom in choosing the detection tool, ...

Soft-Fault Detection Technologies Developed for Electrical Power Systems

Science.gov (United States)

Button, Robert M.

2004-01-01

The NASA Glenn Research Center, partner universities, and defense contractors are working to develop intelligent power management and distribution (PMAD) technologies for future spacecraft and launch vehicles. The goals are to provide higher performance (efficiency, transient response, and stability), higher fault tolerance, and higher reliability through the application of digital control and communication technologies. It is also expected that these technologies will eventually reduce the design, development, manufacturing, and integration costs for large, electrical power systems for space vehicles. The main focus of this research has been to incorporate digital control, communications, and intelligent algorithms into power electronic devices such as direct-current to direct-current (dc-dc) converters and protective switchgear. These technologies, in turn, will enable revolutionary changes in the way electrical power systems are designed, developed, configured, and integrated in aerospace vehicles and satellites. Initial successes in integrating modern, digital controllers have proven that transient response performance can be improved using advanced nonlinear control algorithms. One technology being developed includes the detection of "soft faults," those not typically covered by current systems in use today. Soft faults include arcing faults, corona discharge faults, and undetected leakage currents. Using digital control and advanced signal analysis algorithms, we have shown that it is possible to reliably detect arcing faults in high-voltage dc power distribution systems (see the preceding photograph). Another research effort has shown that low-level leakage faults and cable degradation can be detected by analyzing power system parameters over time. This additional fault detection capability will result in higher reliability for long-lived power systems such as reusable launch vehicles and space exploration missions.

Electrical detection of single magnetic skyrmion at room temperature

Directory of Open Access Journals (Sweden)

Riccardo Tomasello

2017-05-01

Full Text Available This paper proposes a protocol for the electrical detection of a magnetic skyrmion via the change of the tunneling magnetoresistive (TMR signal in a three-terminal device. This approach combines alternating spin-transfer torque from both spin-filtering (due to a perpendicular polarizer and spin-Hall effect with the TMR signal. Micromagnetic simulations, used to test and verify such working principle, show that there exists a frequency region particularly suitable for this achievement. This result can be at the basis of the design of a TMR based read-out for skyrmion detection, overcoming the difficulties introduced by the thermal drift of the skyrmion once nucleated.

Possibilities of microscopic detection of isolated porcine proteins in model meat products

Directory of Open Access Journals (Sweden)

Michaela Petrášová

2016-05-01

Full Text Available In recent years, various protein additives intended for manufacture of meat products have increasing importance in the food industry. These ingredients include both, plant-origin as well as animal-origin proteins. Among animal proteins, blood plasma, milk protein or collagen are used most commonly. Collagen is obtained from pork, beef, and poultry or fish skin. Collagen does not contain all the essential amino acids, thus it is not a full protein in terms of essential amino acids supply for one's organism. However, it is rather rich in amino acids of glycine, hydroxyproline and proline which are almost absent in other proteins and their synthesis is very energy intensive. Collagen, which is added to the soft and small meat products in the form of isolated porcine protein, significantly affects the organoleptic properties of these products. This work focused on detection of isolated porcine protein in model meat products where detection of isolated porcine protein was verified by histological staining and light microscopy. Seven model meat products from poultry meat and 7 model meat products from beef and pork in the ratio of 1:1, which contained 2.5% concentration of various commercially produced isolated porcine proteins, were examined. These model meat products were histologically processed by means of cryosections and stained with hematoxylin-eosin staining, toluidine blue staining and Calleja. For the validation phase, Calleja was utilized. To determine the sensitivity and specificity, five model meat products containing the addition of isolated porcine protein and five model meat products free of it were used. The sensitivity was determined for isolated porcine protein at 1.00 and specificity was determined at 1.00. The detection limit of the method was at the level of 0.001% addition. Repeatability of the method was carried out using products with addition as well as without addition of isolated porcine protein and detection was repeated

Synthetic mRNA devices that detect endogenous proteins and distinguish mammalian cells.

Science.gov (United States)

Kawasaki, Shunsuke; Fujita, Yoshihiko; Nagaike, Takashi; Tomita, Kozo; Saito, Hirohide

2017-07-07

Synthetic biology has great potential for future therapeutic applications including autonomous cell programming through the detection of protein signals and the production of desired outputs. Synthetic RNA devices are promising for this purpose. However, the number of available devices is limited due to the difficulty in the detection of endogenous proteins within a cell. Here, we show a strategy to construct synthetic mRNA devices that detect endogenous proteins in living cells, control translation and distinguish cell types. We engineered protein-binding aptamers that have increased stability in the secondary structures of their active conformation. The designed devices can efficiently respond to target proteins including human LIN28A and U1A proteins, while the original aptamers failed to do so. Moreover, mRNA delivery of an LIN28A-responsive device into human induced pluripotent stem cells (hiPSCs) revealed that we can distinguish living hiPSCs and differentiated cells by quantifying endogenous LIN28A protein expression level. Thus, our endogenous protein-driven RNA devices determine live-cell states and program mammalian cells based on intracellular protein information. © The Author(s) 2017. Published by Oxford University Press on behalf of Nucleic Acids Research.

A real-time insulation detection method for battery packs used in electric vehicles

Science.gov (United States)

Tian, Jiaqiang; Wang, Yujie; Yang, Duo; Zhang, Xu; Chen, Zonghai

2018-05-01

Due to the energy crisis and environmental pollution, electric vehicles have become more and more popular. Compared to traditional fuel vehicles, the electric vehicles are integrated with more high-voltage components, which have potential security risks of insulation. The insulation resistance between the chassis and the direct current bus of the battery pack is easily affected by factors such as temperature, humidity and vibration. In order to ensure the safe and reliable operation of the electric vehicles, it is necessary to detect the insulation resistance of the battery pack. This paper proposes an insulation detection scheme based on low-frequency signal injection method. Considering the insulation detector which can be easily affected by noises, the algorithm based on Kalman filter is proposed. Moreover, the battery pack is always in the states of charging and discharging during driving, which will lead to frequent changes in the voltage of the battery pack and affect the estimation accuracy of insulation detector. Therefore the recursive least squares algorithm is adopted to solve the problem that the detection results of insulation detector mutate with the voltage of the battery pack. The performance of the proposed method is verified by dynamic and static experiments.

Protein remote homology detection based on bidirectional long short-term memory.

Science.gov (United States)

Li, Shumin; Chen, Junjie; Liu, Bin

2017-10-10

Protein remote homology detection plays a vital role in studies of protein structures and functions. Almost all of the traditional machine leaning methods require fixed length features to represent the protein sequences. However, it is never an easy task to extract the discriminative features with limited knowledge of proteins. On the other hand, deep learning technique has demonstrated its advantage in automatically learning representations. It is worthwhile to explore the applications of deep learning techniques to the protein remote homology detection. In this study, we employ the Bidirectional Long Short-Term Memory (BLSTM) to learn effective features from pseudo proteins, also propose a predictor called ProDec-BLSTM: it includes input layer, bidirectional LSTM, time distributed dense layer and output layer. This neural network can automatically extract the discriminative features by using bidirectional LSTM and the time distributed dense layer. Experimental results on a widely-used benchmark dataset show that ProDec-BLSTM outperforms other related methods in terms of both the mean ROC and mean ROC50 scores. This promising result shows that ProDec-BLSTM is a useful tool for protein remote homology detection. Furthermore, the hidden patterns learnt by ProDec-BLSTM can be interpreted and visualized, and therefore, additional useful information can be obtained.

Sensitive targeted multiple protein quantification based on elemental detection of Quantum Dots

Energy Technology Data Exchange (ETDEWEB)

Montoro Bustos, Antonio R.; Garcia-Cortes, Marta [Department of Physical and Analytical Chemistry, University of Oviedo, Julián Clavería 8, Oviedo 33006 (Spain); González-Iglesias, Hector [Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernandez-Vega, Avenida Doctores Fernández-Vega, 34, Oviedo 33012 (Spain); Ruiz Encinar, Jorge, E-mail: ruizjorge@uniovi.es [Department of Physical and Analytical Chemistry, University of Oviedo, Julián Clavería 8, Oviedo 33006 (Spain); Costa-Fernández, José M. [Department of Physical and Analytical Chemistry, University of Oviedo, Julián Clavería 8, Oviedo 33006 (Spain); Coca-Prados, Miguel [Fundación de Investigación Oftalmológica, Instituto Oftalmológico Fernandez-Vega, Avenida Doctores Fernández-Vega, 34, Oviedo 33012 (Spain); Department of Ophthalmology and Visual Science, Yale University School of Medicine, New Haven, CT 06510 (United States); Sanz-Medel, Alfredo, E-mail: asm@uniovi.es [Department of Physical and Analytical Chemistry, University of Oviedo, Julián Clavería 8, Oviedo 33006 (Spain)

2015-06-16

Highlights: • Novel generic platform for multiparametric quantification of proteins. • QDs labeling and ICP-MS detection allow significant analytical signal amplification. • ICP-MS mass balances information provided an internal validation of the immunoassay. • Multiparametric determination of 5 proteins in human serum samples. • ICP-MS reduced matrix effects as compared to other conventional detection techniques. - Abstract: A generic strategy based on the use of CdSe/ZnS Quantum Dots (QDs) as elemental labels for protein quantification, using immunoassays with elemental mass spectrometry (ICP-MS), detection is presented. In this strategy, streptavidin modified QDs (QDs-SA) are bioconjugated to a biotinylated secondary antibody (b-Ab{sub 2}). After a multi-technique characterization of the synthesized generic platform (QDs-SA-b-Ab{sub 2}) it was applied to the sequential quantification of five proteins (transferrin, complement C3, apolipoprotein A1, transthyretin and apolipoprotein A4) at different concentration levels in human serum samples. It is shown how this generic strategy does only require the appropriate unlabeled primary antibody for each protein to be detected. Therefore, it introduces a way out to the need for the cumbersome and specific bioconjugation of the QDs to the corresponding specific recognition antibody for every target analyte (protein). Results obtained were validated with those obtained using UV–vis spectrophotometry and commercial ELISA Kits. As expected, ICP-MS offered one order of magnitude lower DL (0.23 fmol absolute for transferrin) than the classical spectrophotometric detection (3.2 fmol absolute). ICP-MS precision and detection limits, however turned out to be compromised by procedural blanks. The full analytical performance of the ICP-MS-based immunoassay proposed was assessed for detection of transferrin (Tf), present at the low ng mL{sup −1} range in a complex “model” synthetic matrix, where the total protein

Detecting protein complexes based on a combination of topological and biological properties in protein-protein interaction network

Directory of Open Access Journals (Sweden)

Pooja Sharma

2018-06-01

Full Text Available Protein complexes are known to play a major role in controlling cellular activity in a living being. Identifying complexes from raw protein protein interactions (PPIs is an important area of research. Earlier work has been limited mostly to yeast. Such protein complex identification methods, when applied to large human PPIs often give poor performance. We introduce a novel method called CSC to detect protein complexes. The method is evaluated in terms of positive predictive value, sensitivity and accuracy using the datasets of the model organism, yeast and humans. CSC outperforms several other competing algorithms for both organisms. Further, we present a framework to establish the usefulness of CSC in analyzing the influence of a given disease gene in a complex topologically as well as biologically considering eight major association factors. Keywords: Protein complex, Connectivity, Semantic similarity, Contribution

On the detectability of transverse cracks in laminated composites through measurements of electrical potential change

KAUST Repository

Selvakumaran, Lakshmi

2015-01-07

For structures made of laminated composites, real-time structural health monitoring is necessary as significant damage may occur without any visible signs on the surface. Inspection by electrical tomography seems a viable approach as the technique relies on voltage measurements from a network of electrodes over the boundary of the inspected domain to infer the change in conductivity within the bulk material. The change in conductivity, if significant, can be correlated to the degradation state of the material, allowing damage detection. We focus here on the detection of the transverse cracking mechanism which modifies the in-plane transverse conductivity of ply. The quality of detection is directly related to the sensitivity of the voltage measurements with respect to the presence of cracks. We demonstrate here from numerical experiments that the sensitivity depends on several parameters, such as the anisotropy in the electrical conductivity of the baseline composite ply or the geometricalparameters of the structure. Based on these results, applicability of electrical tomography to detect transverse cracks in a laminate is discussed.

Goaf water detection using the grounded electrical source airborne transient electromagnetic system

Science.gov (United States)

Li, D.; Ji, Y.; Guan, S.; Wu, Y.; Wang, A.

2017-12-01

To detect the geoelectric characteristic of goaf water, the grounded electrical source airborne transient electromagnetic (GREATEM) system (developed by Jilin University, China) is applied to the goaf water detection since its advantages of considerable prospecting depth, lateral resolution and detection efficiency. For the test of GREATEM system in goaf water detection, an experimental survey was conducted at Qinshui coal mine (Shanxi province, China). After data acquisition, noise reduction and inversion, the resistivity profiles of survey area is presented. The results highly agree the investigation information provided by Shanxi Coal Geology Geophysical Surveying Exploration Institute (China), conforming that the GREATEM system is an effective technique for resistivity detection of goaf water.

Detection of Intracellular Factor VIII Protein in Peripheral Blood Mononuclear Cells by Flow Cytometry

Directory of Open Access Journals (Sweden)

Gouri Shankar Pandey

2013-01-01

Full Text Available Flow cytometry is widely used in cancer research for diagnosis, detection of minimal residual disease, as well as immune monitoring and profiling following immunotherapy. Detection of specific host proteins for diagnosis predominantly uses quantitative PCR and western blotting assays. In this study, we optimized a flow cytometry-based detection assay for Factor VIII protein in peripheral blood mononuclear cells (PBMCs. An indirect intracellular staining (ICS method was standardized using monoclonal antibodies to different domains of human Factor VIII protein. The FVIII protein expression level was estimated by calculating the mean and median fluorescence intensities (MFI values for each monoclonal antibody. ICS staining of transiently transfected cell lines supported the method's specificity. Intracellular FVIII protein expression was also detected by the monoclonal antibodies used in the study in PBMCs of five blood donors. In summary, our data suggest that intracellular FVIII detection in PBMCs of hemophilia A patients can be a rapid and reliable method to detect intracellular FVIII levels.

Detecting Immune System Response Proteins in a 500 Year-old Inca Mummy

Science.gov (United States)

Corthals, A.; Davalos, L.; Martin, D.W.; Rieger, R.; Chen, E.I.; Koller, A.

2011-01-01

Disease detection in ancient human samples currently relies on genomic-based assays, which are error prone due to contamination and cannot distinguish between active and latent pathogenic infection. On the other hand, protein-based assays such as global protein profiling offer complementary alternatives for the pathological diagnosis of archeological specimen. The discovery of three Inca mummies in 1998, perfectly preserved in the permafrost of the high Andes, allowed us to analyze mummy samples by protein-based and genomic-based assay. A buccal swab from one of the 500 year old mummy was analyzed by shotgun proteomics to detect the protein profile. Among the identified proteins, we found a signature of proteins indicating an immune response to a bacterial infection at the time of the mummy's death. Based on the external visible symptoms and the gamut of immune response proteins obtained from the mouth swab, we suspected that the pulmonary infection was caused by Mycobacterium. PCR assay followed by direct sequencing of the PCR products confirmed the presence of Mycobacterium sp. in the mouth swab. Until now, immunoassays have been the only way to detect an active immune response and infer infection in historical samples, but these were plagued by low specificity and sensitivity. However, we demonstrate here the feasibility of incorporating global protein profiling in the diagnosis of infection from archeological samples. Protein signatures obtained from these samples could be extremely useful in determining the status of infection while genomic-based assays can be used to detect the identity of the pathogen.

A method for detecting hydrophobic patches protein

NARCIS (Netherlands)

Lijnzaad, P.; Berendsen, H.J.C.; Argos, P.

1996-01-01

A method for the detection of hydrophobic patches on the surfaces of protein tertiary structures is presented, it delineates explicit contiguous pieces of surface of arbitrary size and shape that consist solely of carbon and sulphur atoms using a dot representation of the solvent-accessible surface,

Single walled carbon nanotube-based electrical biosensor for the label-free detection of pathogenic bacteria

DEFF Research Database (Denmark)

Yoo, S. M.; Baek, Y. K.; Shin, S.

2016-01-01

We herein describe the development of a single-walled carbon nanotube (SWNT)-based electrical biosensor consisting of a two-terminal resistor, and report its use for the specific, label-free detection of pathogenic bacteria via changes in conductance. The ability of this biosensor to recognize...... different pathogenic bacteria was analyzed, and conditions were optimized with different probe concentrations. Using this system, the reference strains and clinical isolates of Staphylococcus aureus and Escherichia coli were successfully detected; in both cases, the sensor showed a detection limit of 10 CFU....... This SWNT-based electrical biosensor will prove useful for the development of highly sensitive and specific handheld pathogen detectors....

Highly sensitive and specific protein detection via combined capillary isoelectric focusing and proximity ligation

NARCIS (Netherlands)

Padhan, N.; Yan, J.; Boge, A.; Scrivener, E.; Birgisson, H.; Zieba, A.; Gullberg, M.; Kamali-Moghaddam, M.; Claesson-Welsh, L.; Landegren, U.

2017-01-01

Detection and quantification of proteins and their post-translational modifications are crucial to decipher functions of complex protein networks in cell biology and medicine. Capillary isoelectric focusing together with antibody-based detection can resolve and identify proteins and their isoforms

Online slug detection in multi-phase transportation pipelines using electrical tomography

DEFF Research Database (Denmark)

Pedersen, Simon; Mai, Christian; Hansen, Leif

2015-01-01

in the pipelines is a highly investigated topic. To eliminate the slug in an online manner real-time slug detection methods are often required. Traditionally topside pressure transmitters upstream a 3-phase separator have been used as the controlled variable. In this paper Electrical Resistivity Tomography (ERT...

Online Slug Detection in Multi-phase Transportation Pipelines Using Electrical Tomography

DEFF Research Database (Denmark)

Pedersen, Simon; Mai, Christian; Hansen, Leif

2015-01-01

in the pipelines is a highly investigated topic. To eliminate the slug in an online manner real-time slug detection methods are often required. Traditionally topside pressure transmitters upstream a 3-phase separator have been used as the controlled variable. In this paper Electrical Resistivity Tomography (ERT...

Coulomb interactions between cytoplasmic electric fields and phosphorylated messenger proteins optimize information flow in cells.

Directory of Open Access Journals (Sweden)

Robert A Gatenby

2010-08-01

Full Text Available Normal cell function requires timely and accurate transmission of information from receptors on the cell membrane (CM to the nucleus. Movement of messenger proteins in the cytoplasm is thought to be dependent on random walk. However, Brownian motion will disperse messenger proteins throughout the cytosol resulting in slow and highly variable transit times. We propose that a critical component of information transfer is an intracellular electric field generated by distribution of charge on the nuclear membrane (NM. While the latter has been demonstrated experimentally for decades, the role of the consequent electric field has been assumed to be minimal due to a Debye length of about 1 nanometer that results from screening by intracellular Cl- and K+. We propose inclusion of these inorganic ions in the Debye-Huckel equation is incorrect because nuclear pores allow transit through the membrane at a rate far faster than the time to thermodynamic equilibrium. In our model, only the charged, mobile messenger proteins contribute to the Debye length.Using this revised model and published data, we estimate the NM possesses a Debye-Huckel length of a few microns and find this is consistent with recent measurement using intracellular nano-voltmeters. We demonstrate the field will accelerate isolated messenger proteins toward the nucleus through Coulomb interactions with negative charges added by phosphorylation. We calculate transit times as short as 0.01 sec. When large numbers of phosphorylated messenger proteins are generated by increasing concentrations of extracellular ligands, we demonstrate they generate a self-screening environment that regionally attenuates the cytoplasmic field, slowing movement but permitting greater cross talk among pathways. Preliminary experimental results with phosphorylated RAF are consistent with model predictions.This work demonstrates that previously unrecognized Coulomb interactions between phosphorylated messenger

Acquisition of electrical signals using commercial electronic components for detection system of Lead ion in distilled water

Science.gov (United States)

Pujiyanto; Yasin, M.; Rusydi, F.

2018-03-01

Development of lead ion detection systems is expected to have an advantage in terms of simplicity of the device and easy for concentration analysis of a lead ion with very high performance. One important part of lead ion detection systems are electrical signal acquisition parts. The electrical signal acquisition part uses the main electronic components: non inverting op-amplifier, instrumentation amplifier, multiplier circuit and logarithmic amplifier. Here will be shown the performance of lead ion detection systems when the existing electrical signal processors use commercial electronic components. The results that can be drawn from this experimental were the lead ion sensor that has been developed can be used to detect lead ions with a sensitivity of 10.48 mV/ppm with the linearity 97.11% and had a measurement range of 0.1 ppm to 80 ppm.

Two-photon excited UV fluorescence for protein crystal detection

International Nuclear Information System (INIS)

Madden, Jeremy T.; DeWalt, Emma L.; Simpson, Garth J.

2011-01-01

Complementary measurements using SONICC and TPE-UVF allow the sensitive and selective detection of protein crystals. Two-photon excited ultraviolet fluorescence (TPE-UVF) microscopy is explored for sensitive protein-crystal detection as a complement to second-order nonlinear optical imaging of chiral crystals (SONICC). Like conventional ultraviolet fluorescence (UVF), TPE-UVF generates image contrast based on the intrinsic fluorescence of aromatic residues, generally producing higher fluorescence emission within crystals than the mother liquor by nature of the higher local protein concentration. However, TPE-UVF has several advantages over conventional UVF, including (i) insensitivity to optical scattering, allowing imaging in turbid matrices, (ii) direct compatibility with conventional optical plates and windows by using visible light for excitation, (iii) elimination of potentially damaging out-of-plane UV excitation, (iv) improved signal to noise through background reduction from out-of-plane excitation and (v) relatively simple integration into instrumentation developed for SONICC

Method and system for early detection of incipient faults in electric motors

Science.gov (United States)

Parlos, Alexander G; Kim, Kyusung

2003-07-08

A method and system for early detection of incipient faults in an electric motor are disclosed. First, current and voltage values for one or more phases of the electric motor are measured during motor operations. A set of current predictions is then determined via a neural network-based current predictor based on the measured voltage values and an estimate of motor speed values of the electric motor. Next, a set of residuals is generated by combining the set of current predictions with the measured current values. A set of fault indicators is subsequently computed from the set of residuals and the measured current values. Finally, a determination is made as to whether or not there is an incipient electrical, mechanical, and/or electromechanical fault occurring based on the comparison result of the set of fault indicators and a set of predetermined baseline values.

Detection of proteins on blot transfer membranes.

Science.gov (United States)

Sasse, Joachim; Gallagher, Sean R

2003-11-01

In the basic and alternate protocols of this unit, proteins are stained after electroblotting from polyacrylamide gels to blot transfer membranes. If the samples of interest are electrophoresed in duplicate and transferred to a blot transfer membrane, half of the membrane can be stained to determine the efficiency of transfer to the membrane and the other half can be used for immunoblotting (i.e., western blotting). Detection limits of each staining method are given along with a list of compatible blot transfer membranes and gels. A support protocol describes a method for alkali treatment that enhances subsequent staining of bound proteins.

Development of optical immunosensors for detection of proteins in serum.

Science.gov (United States)

Kyprianou, Dimitris; Chianella, Iva; Guerreiro, Antonio; Piletska, Elena V; Piletsky, Sergey A

2013-01-15

The detection of proteins in biological samples such as blood, serum or plasma by biosensors is very challenging due to the complex nature of the matrix, which contains a high level of many interfering compounds. Here we show the application of a novel polymeric immobilisation matrix that helps in the detection of specific protein analytes in biological samples by surface plasmon resonance (SPR) immunosensors. This polymer matrix contains thioacetal functional groups included in the network, and these groups do not require any further activation in order to react with proteins, making it attractive for sensor fabrication. The protein prostate specific antigen (PSA) was selected as a model target analyte. A sandwich format with two primary antibodies recognising different parts (epitopes) of the analyte was used for the detection of PSA in serum. The efficiency of the reduction of non-specific binding achieved with novel polymer was compared with those of other techniques such as coating of sensor surface with polyethylene glycol (PEG), use of charged hydrophilic aspartic acid and surfactants such as Tween20. The detection limit of the polymer based immunosensor was 0.1 ng ml(-1) for free form PSA (f-PSA) in buffer and 5 ng ml(-1) in 20% serum. This is an improvement compared with similar devices reported on literature, indicating the potential of the immunosensor developed here for the analysis of real samples. Copyright © 2012 Elsevier B.V. All rights reserved.

Label-Free Electrical Detection Using Carbon Nanotube-Based Biosensors

Directory of Open Access Journals (Sweden)

Kenzo Maehashi

2009-07-01

Full Text Available Label-free detections of biomolecules have attracted great attention in a lot of life science fields such as genomics, clinical diagnosis and practical pharmacy. In this article, we reviewed amperometric and potentiometric biosensors based on carbon nanotubes (CNTs. In amperometric detections, CNT-modified electrodes were used as working electrodes to significantly enhance electroactive surface area. In contrast, the potentiometric biosensors were based on aptamer-modified CNT field-effect transistors (CNTFETs. Since aptamers are artificial oligonucleotides and thus are smaller than the Debye length, proteins can be detected with high sensitivity. In this review, we discussed on the technology, characteristics and developments for commercialization in label-free CNT-based biosensors.

Laser cooling and optical detection of excitations in a LC electrical circuit

DEFF Research Database (Denmark)

Taylor, J. M.; Sørensen, Anders Søndberg; Marcus, Charles Masamed

2011-01-01

We explore a method for laser cooling and optical detection of excitations in a room temperature LC electrical circuit. Our approach uses a nanomechanical oscillator as a transducer between optical and electronic excitations. An experimentally feasible system with the oscillator capacitively...... coupled to the LC and at the same time interacting with light via an optomechanical force is shown to provide strong electromechanical coupling. Conditions for improved sensitivity and quantum limited readout of electrical signals with such an “optical loud speaker” are outlined....

Automatic Detection of Electric Power Troubles (ADEPT)

Science.gov (United States)

Wang, Caroline; Zeanah, Hugh; Anderson, Audie; Patrick, Clint; Brady, Mike; Ford, Donnie

1988-11-01

Automatic Detection of Electric Power Troubles (A DEPT) is an expert system that integrates knowledge from three different suppliers to offer an advanced fault-detection system. It is designed for two modes of operation: real time fault isolation and simulated modeling. Real time fault isolation of components is accomplished on a power system breadboard through the Fault Isolation Expert System (FIES II) interface with a rule system developed in-house. Faults are quickly detected and displayed and the rules and chain of reasoning optionally provided on a laser printer. This system consists of a simulated space station power module using direct-current power supplies for solar arrays on three power buses. For tests of the system's ablilty to locate faults inserted via switches, loads are configured by an INTEL microcomputer and the Symbolics artificial intelligence development system. As these loads are resistive in nature, Ohm's Law is used as the basis for rules by which faults are located. The three-bus system can correct faults automatically where there is a surplus of power available on any of the three buses. Techniques developed and used can be applied readily to other control systems requiring rapid intelligent decisions. Simulated modeling, used for theoretical studies, is implemented using a modified version of Kennedy Space Center's KATE (Knowledge-Based Automatic Test Equipment), FIES II windowing, and an ADEPT knowledge base.

Manufacturing and testing flexible microfluidic devices with optical and electrical detection mechanisms

NARCIS (Netherlands)

Ivan, M.G.; Vivet, F.; Meinders, E.R.

2010-01-01

Flexible microfluidic devices made of poly(dimethylsiloxane) (PDMS) were manufactured by soft lithography, and tested in detection of ionic species using optical absorption spectroscopy and electrical measurements. PDMS was chosen due to its flexibility and ease of surface modification by exposure

MOLECULAR DOCKING AND DYNAMICS STUDIES ON THE PROTEIN-PROTEIN INTERACTIONS OF ELECTRICALLY ACTIVE PILIN NANOWIRES OF GEOBACTER SULFURREDUCENS.

Directory of Open Access Journals (Sweden)

D. Jeya Sundara Sharmila1 *

2017-06-01

Full Text Available Molecular interactions are key aspects in biological recognitions applicable in nano/micro systems. Bacterial nanowires are pilus filament based structures that can conduct electrons. The transport of electron is proposed to be facilitated by filamentous fibers made up of polymeric assemblies of proteins called pilin. Geobacter sulfurreducens is capable of delivering electrons through extracellular electron transport (EET by employing conductive nanowires, which are pilin proteins composed of type IV subunit PilA. Protein-protein interactions play an important role in the stabilization of the pilin nanowire assembly complex and it contains transmembrane (TM domain. In current study, protein-protein docking and multiple molecular dynamic (MD simulations were performed to understand the binding mode of pilin nanowires. The MD result explains the conformational behavior and folding of pilin nanowires in water environment in different time scale duration 20, 5, 5, 10 and 20ns (total of 60ns. Direct hydrogen bonds and water mediated hydrogen bonds that play a crucial role during the simulation were investigated. The conformational state, folding, end-toend distance profile and hydrogen bonding behavior had indicated that the Geobacter sulfurreducens pilin nanowires have electrical conductivity properties.

Detection of Alkylating Agents using Electrical and Mechanical Means

Science.gov (United States)

Gerchikov, Yulia; Borzin, Elena; Gannot, Yair; Shemesh, Ariel; Meltzman, Shai; Hertzog-Ronen, Carmit; Tal, Shay; Stolyarova, Sara; Nemirovsky, Yael; Tessler, Nir; Eichen, Yoav

2011-08-01

Alkylating agents are reactive molecules having at least one polar bond between a carbon atom and a good leaving group. These often simple molecules are frequently used in organic synthesis, as sterilizing agents in agriculture and even as anticancer agents in medicine. Unfortunately, for over a century, some of the highly reactive alkylating agents are also being used as blister chemical warfare agents. Being relatively simple to make, the risk is that these will be applied by terrorists as poor people warfare agents. The detection and identification of such alkylating agents is not a simple task because of their high reactivity and simple structure of the reactive site. Here we report on new approaches to the detection and identification of such alkylating agents using electrical (organic field effect transistors) and mechanical (microcantilevers) means.

Detection of Alkylating Agents using Electrical and Mechanical Means

International Nuclear Information System (INIS)

Gerchikov, Yulia; Borzin, Elena; Gannot, Yair; Shemesh, Ariel; Meltzman, Shai; Hertzog-Ronen, Carmit; Eichen, Yoav; Tal, Shay; Stolyarova, Sara; Nemirovsky, Yael; Tessler, Nir

2011-01-01

Alkylating agents are reactive molecules having at least one polar bond between a carbon atom and a good leaving group. These often simple molecules are frequently used in organic synthesis, as sterilizing agents in agriculture and even as anticancer agents in medicine. Unfortunately, for over a century, some of the highly reactive alkylating agents are also being used as blister chemical warfare agents. Being relatively simple to make, the risk is that these will be applied by terrorists as poor people warfare agents. The detection and identification of such alkylating agents is not a simple task because of their high reactivity and simple structure of the reactive site. Here we report on new approaches to the detection and identification of such alkylating agents using electrical (organic field effect transistors) and mechanical (microcantilevers) means.

Detection of Alkylating Agents using Electrical and Mechanical Means

Energy Technology Data Exchange (ETDEWEB)

Gerchikov, Yulia; Borzin, Elena; Gannot, Yair; Shemesh, Ariel; Meltzman, Shai; Hertzog-Ronen, Carmit; Eichen, Yoav [Schulich Department of Chemistry, Technion-Israel Institute of Technology, Technion City, 32000, Haifa (Israel) (Israel); Tal, Shay [Present address: Systems Biology Department, Harvard Medical School, Boston, MA 02115 (United States); Stolyarova, Sara; Nemirovsky, Yael; Tessler, Nir, E-mail: chryoav@tx.technion.ac.il [Department of Electrical Engineering, Technion-Israel Institute of Technology, Technion City, 32000, Haifa (Israel)

2011-08-17

Alkylating agents are reactive molecules having at least one polar bond between a carbon atom and a good leaving group. These often simple molecules are frequently used in organic synthesis, as sterilizing agents in agriculture and even as anticancer agents in medicine. Unfortunately, for over a century, some of the highly reactive alkylating agents are also being used as blister chemical warfare agents. Being relatively simple to make, the risk is that these will be applied by terrorists as poor people warfare agents. The detection and identification of such alkylating agents is not a simple task because of their high reactivity and simple structure of the reactive site. Here we report on new approaches to the detection and identification of such alkylating agents using electrical (organic field effect transistors) and mechanical (microcantilevers) means.

Highly sensitive rapid fluorescence detection of protein residues on surgical instruments

International Nuclear Information System (INIS)

Kovalev, Valeri I; Bartona, James S; Richardson, Patricia R; Jones, Anita C

2006-01-01

There is a risk of contamination of surgical instruments by infectious protein residues, in particular, prions which are the agents for Creutzfeldt-Jakob Disease in humans. They are exceptionally resistant to conventional sterilization, therefore it is important to detect their presence as contaminants so that alternative cleaning procedures can be applied. We describe the development of an optimized detection system for fluorescently labelled protein, suitable for in-hospital use. We show that under optimum conditions the technique can detect ∼10 attomole/cm 2 with a scan speed of ∼3-10 cm 2 /s of the test instrument's surface. A theoretical analysis and experimental measurements will be discussed

Optimal combinations of acute phase proteins for detecting infectious disease in pigs

DEFF Research Database (Denmark)

Heegaard, Peter M. H.; Stockmarr, Anders; Piñeiro, Matilde

2011-01-01

The acute phase protein (APP) response is an early systemic sign of disease, detected as substantial changes in APP serum concentrations and most disease states involving inflammatory reactions give rise to APP responses. To obtain a detailed picture of the general utility of porcine APPs to detect...... gondii) and one viral (porcine respiratory and reproductive syndrome virus) infection and one aseptic inflammation. Immunochemical analyses of seven APPs, four positive (C-reactive protein (CRP), haptoglobin (Hp), pig major acute phase protein (pigMAP) and serum amyloid A (SAA)) and three negative...

Enhancement of delay eyelid conditioning by microcurrent electrical stimulation of the medial prefrontal cortex is triggered by the expression of Fos protein in guinea pigs.

Science.gov (United States)

Zheng, Ya-Juan; Dong, Yu-Chen; Zhu, Chao; Zhao, Mei-Sheng

2016-03-01

Eyelid conditioning, including delay eyelid conditioning and trace eyelid conditioning, has been used extensively to study neural structures and mechanisms of learning and memory as a form of associative learning. In the present study, microcurrent electrical stimulation was used to stimulate the medial prefrontal cortex (mPFC) to induce delay eyelid conditioning in guinea pigs. The acquisition rate and relative latency of the conditioned eyelid response (CR) and the startle eyelid response (SR) were analyzed. The mPFC sites in the guinea pigs were examined under a light microscope following Nissl staining. In addition, the expression of Fos protein in neurons was detected using immunohistochemistry and western blot analysis. The results indicated that the acquisition rates of CR and SR were increased significantly (Pmicrocurrent electrical stimulation of the mPFC in guinea pigs was triggered by the expression of Fos protein. The observations of the present study further expand the understanding of conditioned reflexes and may aid future investigations into the formation of eyelid conditioning and the mechanisms underlying the circuit in various conditions.

Protein complex detection in PPI networks based on data integration and supervised learning method.

Science.gov (United States)

Yu, Feng; Yang, Zhi; Hu, Xiao; Sun, Yuan; Lin, Hong; Wang, Jian

2015-01-01

Revealing protein complexes are important for understanding principles of cellular organization and function. High-throughput experimental techniques have produced a large amount of protein interactions, which makes it possible to predict protein complexes from protein-protein interaction (PPI) networks. However, the small amount of known physical interactions may limit protein complex detection. The new PPI networks are constructed by integrating PPI datasets with the large and readily available PPI data from biomedical literature, and then the less reliable PPI between two proteins are filtered out based on semantic similarity and topological similarity of the two proteins. Finally, the supervised learning protein complex detection (SLPC), which can make full use of the information of available known complexes, is applied to detect protein complex on the new PPI networks. The experimental results of SLPC on two different categories yeast PPI networks demonstrate effectiveness of the approach: compared with the original PPI networks, the best average improvements of 4.76, 6.81 and 15.75 percentage units in the F-score, accuracy and maximum matching ratio (MMR) are achieved respectively; compared with the denoising PPI networks, the best average improvements of 3.91, 4.61 and 12.10 percentage units in the F-score, accuracy and MMR are achieved respectively; compared with ClusterONE, the start-of the-art complex detection method, on the denoising extended PPI networks, the average improvements of 26.02 and 22.40 percentage units in the F-score and MMR are achieved respectively. The experimental results show that the performances of SLPC have a large improvement through integration of new receivable PPI data from biomedical literature into original PPI networks and denoising PPI networks. In addition, our protein complexes detection method can achieve better performance than ClusterONE.

Highly increased detection of silver stained protein bands in polyacrylamide gels with thermo-optical methods

Science.gov (United States)

Mazza, Giulia; Posnicek, Thomas; Brandl, Martin

2016-11-01

Sodium dodecyl sulfate polyacrylamide gel electrophoresis is a well-known technique to separate proteins by their molecular weight. After electrophoresis, the gels are commonly stained for protein band analysis with silver stain; this allows the detection of protein loads to about 1 ng. To increase the detection sensitivity of the protein bands down in the subnanogram level, a sensor has been developed based on the thermal lens effect to scan and quantify protein loads which would remain undetected using the standard imaging systems. The thermal lens sensor is equipped with a 450 nm diode pump laser modulated at 1 Hz and a HeNe probe laser mounted in collinear geometry. The sensor could detect protein bands of 0.05 ng when the gel was soaked in methanol/water and 0.1 ng in water. The limit of detection ranged from 8 to 20 pg, depending on the soaking medium and the staining efficiency. Thus, the detection of silver stain by thermal lens effect results 10 to 20 times more sensitive than the standard colorimetric method.

On the detectability of transverse cracks in laminated composites using electrical potential change measurements

KAUST Repository

Selvakumaran, Lakshmi

2015-03-01

Real-time health monitoring of structures made of laminated composites is necessary as significant damage may occur without any visible signs on the surface. Inspection by electrical tomography (ET) seems a viable approach that relies on voltage measurements from a network of electrodes across the inspected domain to infer conductivity change within the bulk material. If conductivity decreases significantly with increasing damage, the obtained conductivity map can be correlated to the degradation state of the material. We focus here on detection of transverse cracks. As transverse cracks modify the in-plane transverse conductivity of a single ply, we expect them to be detectable by electrical measurements. Yet, the quality of detection is directly related to the sensitivity of the measurements to the presence of cracks. We use numerical experiments to demonstrate that the sensitivity depends on several material and geometrical parameters. Based on the results, the applicability of ET to detect transverse cracks is discussed. One conclusion from the study is that detecting transverse cracks using ET is more reliable in some laminate configurations than in others. Recommendations about the properties of either the pristine material or the inspected structures are provided to establish if ET is reliable in detecting transverse cracks.

Simplified Swarm Optimization-Based Function Module Detection in Protein–Protein Interaction Networks

Directory of Open Access Journals (Sweden)

Xianghan Zheng

2017-04-01

Full Text Available Proteomics research has become one of the most important topics in the field of life science and natural science. At present, research on protein–protein interaction networks (PPIN mainly focuses on detecting protein complexes or function modules. However, existing approaches are either ineffective or incomplete. In this paper, we investigate detection mechanisms of functional modules in PPIN, including open database, existing detection algorithms, and recent solutions. After that, we describe the proposed approach based on the simplified swarm optimization (SSO algorithm and the knowledge of Gene Ontology (GO. The proposed solution implements the SSO algorithm for clustering proteins with similar function, and imports biological gene ontology knowledge for further identifying function complexes and improving detection accuracy. Furthermore, we use four different categories of species datasets for experiment: fruitfly, mouse, scere, and human. The testing and analysis result show that the proposed solution is feasible, efficient, and could achieve a higher accuracy of prediction than existing approaches.

Optimization of proximity ligation assay (PLA) for detection of protein interactions and fusion proteins in non-adherent cells: application to pre-B lymphocytes.

Science.gov (United States)

Debaize, Lydie; Jakobczyk, Hélène; Rio, Anne-Gaëlle; Gandemer, Virginie; Troadec, Marie-Bérengère

2017-01-01

Genetic abnormalities, including chromosomal translocations, are described for many hematological malignancies. From the clinical perspective, detection of chromosomal abnormalities is relevant not only for diagnostic and treatment purposes but also for prognostic risk assessment. From the translational research perspective, the identification of fusion proteins and protein interactions has allowed crucial breakthroughs in understanding the pathogenesis of malignancies and consequently major achievements in targeted therapy. We describe the optimization of the Proximity Ligation Assay (PLA) to ascertain the presence of fusion proteins, and protein interactions in non-adherent pre-B cells. PLA is an innovative method of protein-protein colocalization detection by molecular biology that combines the advantages of microscopy with the advantages of molecular biology precision, enabling detection of protein proximity theoretically ranging from 0 to 40 nm. We propose an optimized PLA procedure. We overcome the issue of maintaining non-adherent hematological cells by traditional cytocentrifugation and optimized buffers, by changing incubation times, and modifying washing steps. Further, we provide convincing negative and positive controls, and demonstrate that optimized PLA procedure is sensitive to total protein level. The optimized PLA procedure allows the detection of fusion proteins and protein interactions on non-adherent cells. The optimized PLA procedure described here can be readily applied to various non-adherent hematological cells, from cell lines to patients' cells. The optimized PLA protocol enables detection of fusion proteins and their subcellular expression, and protein interactions in non-adherent cells. Therefore, the optimized PLA protocol provides a new tool that can be adopted in a wide range of applications in the biological field.

Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

Energy Technology Data Exchange (ETDEWEB)

Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan); Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki [System Instruments Co., Ltd., 776-2 Komiya-cho, Hachioji, Tokyo 192-0031 (Japan); Noda, Mamoru; Igimi, Shizunobu [Division of Biomedical Food Research, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo 158-8501 (Japan); Ikebukuro, Kazunori, E-mail: ikebu@cc.tuat.ac.jp [Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588 (Japan)

2013-11-01

Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10{sup 6} copies.

Automatic polymerase chain reaction product detection system for food safety monitoring using zinc finger protein fused to luciferase

International Nuclear Information System (INIS)

Yoshida, Wataru; Kezuka, Aki; Murakami, Yoshiyuki; Lee, Jinhee; Abe, Koichi; Motoki, Hiroaki; Matsuo, Takafumi; Shimura, Nobuaki; Noda, Mamoru; Igimi, Shizunobu; Ikebukuro, Kazunori

2013-01-01

Graphical abstract: -- Highlights: •Zif268 fused to luciferase was used for E. coli O157, Salmonella and coliform detection. •Artificial zinc finger protein fused to luciferase was constructed for Norovirus detection. •An analyzer that automatically detects PCR products by zinc finger protein fused to luciferase was developed. •Target pathogens were specifically detected by the automatic analyzer with zinc finger protein fused to luciferase. -- Abstract: An automatic polymerase chain reaction (PCR) product detection system for food safety monitoring using zinc finger (ZF) protein fused to luciferase was developed. ZF protein fused to luciferase specifically binds to target double stranded DNA sequence and has luciferase enzymatic activity. Therefore, PCR products that comprise ZF protein recognition sequence can be detected by measuring the luciferase activity of the fusion protein. We previously reported that PCR products from Legionella pneumophila and Escherichia coli (E. coli) O157 genomic DNA were detected by Zif268, a natural ZF protein, fused to luciferase. In this study, Zif268–luciferase was applied to detect the presence of Salmonella and coliforms. Moreover, an artificial zinc finger protein (B2) fused to luciferase was constructed for a Norovirus detection system. In the luciferase activity detection assay, several bound/free separation process is required. Therefore, an analyzer that automatically performed the bound/free separation process was developed to detect PCR products using the ZF–luciferase fusion protein. By means of the automatic analyzer with ZF–luciferase fusion protein, target pathogenic genomes were specifically detected in the presence of other pathogenic genomes. Moreover, we succeeded in the detection of 10 copies of E. coli BL21 without extraction of genomic DNA by the automatic analyzer and E. coli was detected with a logarithmic dependency in the range of 1.0 × 10 to 1.0 × 10 6 copies

Highly sensitive rapid fluorescence detection of protein residues on surgical instruments

Energy Technology Data Exchange (ETDEWEB)

Kovalev, Valeri I [School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS (United Kingdom); Bartona, James S [School of Engineering and Physical Sciences, Heriot-Watt University, Edinburgh EH14 4AS (United Kingdom); Richardson, Patricia R [School of Chemistry, University of Edinburgh, Edinburgh, EH9 3JJ (United Kingdom); Jones, Anita C [School of Chemistry, University of Edinburgh, Edinburgh, EH9 3JJ (United Kingdom)

2006-07-15

There is a risk of contamination of surgical instruments by infectious protein residues, in particular, prions which are the agents for Creutzfeldt-Jakob Disease in humans. They are exceptionally resistant to conventional sterilization, therefore it is important to detect their presence as contaminants so that alternative cleaning procedures can be applied. We describe the development of an optimized detection system for fluorescently labelled protein, suitable for in-hospital use. We show that under optimum conditions the technique can detect {approx}10 attomole/cm{sup 2} with a scan speed of {approx}3-10 cm{sup 2}/s of the test instrument's surface. A theoretical analysis and experimental measurements will be discussed.

A simple and effective method for detecting precipitated proteins in MALDI-TOF MS.

Science.gov (United States)

Oshikane, Hiroyuki; Watabe, Masahiko; Nakaki, Toshio

2018-04-01

MALDI-TOF MS has developed rapidly into an essential analytical tool for the life sciences. Cinnamic acid derivatives are generally employed in routine molecular weight determinations of intact proteins using MALDI-TOF MS. However, a protein of interest may precipitate when mixed with matrix solution, perhaps preventing MS detection. We herein provide a simple approach to enable the MS detection of such precipitated protein species by means of a "direct deposition method" -- loading the precipitant directly onto the sample plate. It is thus expected to improve routine MS analysis of intact proteins. Copyright © 2018. Published by Elsevier Inc.

Protein A Detection Based on Quantum Dots-Antibody Bioprobe Using Fluorescence Coupled Capillary Electrophoresis

Directory of Open Access Journals (Sweden)

Lin Qiu

2014-01-01

Full Text Available In this report, fluorescence detection coupled capillary electrophoresis (CE-FL was used to detect Protein A. Antibody was first labeled with Cy5 and then mixed with quantum dots (QDs to form QDs-antibody bioprobe. Further, we observed fluorescence resonance energy transfer (FRET from QDs donor to Cy5 acceptor. The bioprobe was formed and brought QDs and Cy5 close enough to allow FRET to occur. After adding protein A, the FRET system was broken and caused the FRET signal to decrease. Thus, a new method for the determination of protein A was proposed based on the FRET signal changes. This study provides a new trail of thought for the detection of protein.

Ferritin protein imaging and detection by magnetic force microscopy.

Science.gov (United States)

Hsieh, Chiung-Wen; Zheng, Bin; Hsieh, Shuchen

2010-03-14

Magnetic force microscopy was used to image and detect ferritin proteins and the strength of the magnetic signal is discussed, revealing a large workable lift height between the magnetic tip and the ferritin sample.

Fast electric field waveforms and near-surface electric field images of lightning discharges detected on Mt. Aragats in Armenia

International Nuclear Information System (INIS)

Chilingarian, A.; Khanikyants, Y.; Kozliner, L.; Soghomonyan, S.

2016-01-01

We present the observational data on fast electric waveforms that are detected at 3200 m altitudes above sea level on Mt. Aragats in Armenia during thunderstorms. We analyse the relations of these forms with count rates of particle flux (during Thunderstorm Ground Enhancements -TGEs); to the slow disturbance of the near-surface electrostatic field; and to the lightning location data from the World Wide Lightning Location Network (WWLLN). An observed negative lightning that decreases a negative charge overhead often abruptly terminates TGEs. By analysing the recorded fast electric field waveforms and comparing them with similar classified waveforms reported previously, we could identify the type and polarity of the observed lightnings. (author)

System for detecting and limiting electrical ground faults within electrical devices

International Nuclear Information System (INIS)

Gaubatz, D.C.

1990-01-01

This paper discusses, in a nuclear power plant of a variety wherein a reactor is provided including a reactor vessel retaining a liquid metal coolant, a reactor core and an electromagnetic pump having inductive windings insulatively retained within the electrically conductive wall of an enclosure, the method for controlling electrical ground fault current between a the inductive winding and the walls. It comprises providing an electrically isolated power source by inductive coupling with the plant power supply; rectifying the power source to provide an isolated d.c. power source; providing an inverter powered from the isolated d.c. power source under the control of the plant control system for selectively energizing the inductive windings; providing a fault control conductor electrically connected with the pump enclosure wall and extending as an electrical return for ground fault current to the inverter; and providing an electrical resistance between the conductor and the isolated inverter having an impedance selected to limit the fault current below a predetermined value limiting arc damage at any the electrical ground fault location

Laser-based optical activity detection of amino acids and proteins

Energy Technology Data Exchange (ETDEWEB)

Reitsma, B.H.

1987-01-01

The optical activity detector (OAD) for HPLC is a selective detector for optically active substances including amino acids and proteins. This study illustrates the use of the OAD in three related areas. Section I illustrates the separation of four free amino acids using cation-exchange chromatography. Detection by coupling the OAD to a refractive index detector (RI) for proline and threonine and the OAD to an ultraviolet absorbance detector (UV) for tyrosine and phenylalanine allows the calculation of enantiomeric (D/L) ratios of these amino acids without physical separation. Specific rotations of these four amino acids are also reported. Section II illustrates the separation of 16 dansyl-L-amino acids by RP-HPLC with detection by OA/UV. Section III illustrates the RP-HPLC separation of conformers of soybean trypsin inhibitor. Detection by OA/UV provides insights from the chromatogram unavailable for UV absorbance detection alone. In addition, identification of impurities is simplified with OA/UV. Specific rotations of the separated protein fractions show no significant change accompanying change in conformation.

Electrophoretic Detection and Confocal Microscopic Imaging of Tyrosine Nitrated Proteins in Plant Tissue.

Science.gov (United States)

Arora, Dhara; Singh, Neha; Bhatla, Satish C

2018-01-01

Tyrosine nitrated proteins can be detected in plant cells electrophoretically and their distribution can be monitored by confocal laser scanning microscopy (CLSM) imaging. One-dimensional polyacrylamide gel electrophoresis (1D PAGE) followed by Western blotting using polyclonal antibody against 3-nitrotyrosine residues enables detection of tyrosine nitrated proteins in plant cells. Here we describe detection of tyrosine nitrated proteins in the homogenates derived from sunflower (Helianthus annuus L.) seedling cotyledons. Total soluble proteins obtained from tissue homogenates are resolved using vertical gel electrophoresis followed by their electrophoretic transfer on to a microporous membrane support for immunodetection. Spatial distribution of tyrosine nitrated proteins can be visualized using an antibody against 3-nitrotyrosine residues. Immunofluorescent localization is performed by cutting 7 μm thick wax sections of tissue followed by incubation in primary anti-nitrotyrosine antibody (dilution 1:200) and secondary Cy-3 labeled anti-rabbit IgG antibody (dilution 1:1500). Confocal laser scanning microscopy analysis is undertaken using argon lasers (ex: 530-550 nm and em: 570 nm) at pinhole 1. Modulation in the abundance and spatial localization of tyrosine nitrated proteins in plant tissues can be monitored using these techniques.

Specific detection of proteins using Nanomechanical resonators

DEFF Research Database (Denmark)

Fischer, Lee MacKenzie; Wright, V.A.; Guthy, C.

2008-01-01

of probes onto their surfaces in order to enable the specificity of the detection. Such nanoresonator-based specific detection of proteins is here reported using streptavidin as target system, and immobilized biotin as probe. Nanomechanical resonators resistant to stiction were first realized from silicon...... carbonitride using a novel fabrication method. Vapor-phase deposition of mercaptopropyl trimethoxysilane was performed, and an added mass of 2.22 +/- 0.07 fg/mu m(2) was measured. This linker molecule was used to attach biotin onto the devices, enabling the specific detection of streptavidin. A mass of 3.6 fg....../mu m(2) was attributed to the added streptavidin, corresponding to one molecule per 27 nm(2). The specificity of this recognition was confirmed by exposing the devices to a solution of streptavidin that was already saturated with biotin. An additional negative control was also performed by also...

Detection of site specific glycosylation in proteins using flow cytometry†

Science.gov (United States)

Jayakumar, Deepak; Marathe, Dhananjay D.; Neelamegham, Sriram

2009-01-01

We tested the possibility that it is possible to express unique peptide probes on cell surfaces and detect site-specific glycosylation on these peptides using flow cytometry. Such development can enhance the application of flow cytometry to detect and quantify post-translational modifications in proteins. To this end, the N-terminal section of the human leukocyte glycoprotein PSGL-1 (P-selectin glycoprotein ligand-1) was modified to contain a poly-histidine tag followed by a proteolytic cleavage site. Amino acids preceding the cleavage site have a single O-linked glycosylation site. The recombinant protein called PSGL-1 (HT) was expressed on the surface of two mammalian cell lines, CHO and HL-60, using a lentiviral delivery approach. Results demonstrate that the N-terminal portion of PSGL-1 (HT) can be released from these cells by protease, and the resulting peptide can be readily captured and detected using cytometry-bead assays. Using this strategy, the peptide was immunoprecipitated onto beads bearing mAbs against either the poly-histidine sequence or the human PSGL-1. The carbohydrate epitope associated with the released peptide was detected using HECA-452 and CSLEX-1, monoclonal antibodies that recognize the sialyl Lewis-X epitope. Finally, the peptide released from cells could be separated and enriched using nickel chelate beads. Overall, such an approach that combines recombinant protein expression with flow cytometry, may be useful to quantify changes in site-specific glycosylation for basic science and clinical applications. PMID:19735085

Proton detection for signal enhancement in solid-state NMR experiments on mobile species in membrane proteins

Energy Technology Data Exchange (ETDEWEB)

Ward, Meaghan E.; Ritz, Emily [University of Guelph, Department of Physics (Canada); Ahmed, Mumdooh A. M. [Suez University, The Department of Physics, Faculty of Science (Egypt); Bamm, Vladimir V.; Harauz, George [University of Guelph, Biophysics Interdepartmental Group (Canada); Brown, Leonid S.; Ladizhansky, Vladimir, E-mail: vladizha@uoguelph.ca [University of Guelph, Department of Physics (Canada)

2015-12-15

Direct proton detection is becoming an increasingly popular method for enhancing sensitivity in solid-state nuclear magnetic resonance spectroscopy. Generally, these experiments require extensive deuteration of the protein, fast magic angle spinning (MAS), or a combination of both. Here, we implement direct proton detection to selectively observe the mobile entities in fully-protonated membrane proteins at moderate MAS frequencies. We demonstrate this method on two proteins that exhibit different motional regimes. Myelin basic protein is an intrinsically-disordered, peripherally membrane-associated protein that is highly flexible, whereas Anabaena sensory rhodopsin is composed of seven rigid transmembrane α-helices connected by mobile loop regions. In both cases, we observe narrow proton linewidths and, on average, a 10× increase in sensitivity in 2D insensitive nuclear enhancement of polarization transfer-based HSQC experiments when proton detection is compared to carbon detection. We further show that our proton-detected experiments can be easily extended to three dimensions and used to build complete amino acid systems, including sidechain protons, and obtain inter-residue correlations. Additionally, we detect signals which do not correspond to amino acids, but rather to lipids and/or carbohydrates which interact strongly with membrane proteins.

Bovine Pancreatic Trypsin Inhibitor-Trypsin Complex as a Detection System for Recombinant Proteins

Science.gov (United States)

Borjigin, Jimo; Nathans, Jeremy

1993-01-01

Bovine pancreatic trypsin inhibitor (BPTI) binds to trypsin and anhydrotrypsin (an enzymatically inactive derivative of trypsin) with affinities of 6 x 10-14 and 1.1 x 10-13 M, respectively. We have taken advantage of the high affinity and specificity of this binding reaction to develop a protein tagging system in which biotinylated trypsin or biotinylated anhydrotrypsin is used as the reagent to detect recombinant fusion proteins into which BPTI has been inserted. Two proteins, opsin and growth hormone, were used as targets for insertional mutagenesis with BPTI. In each case, both domains of the fusion protein appear to be correctly folded. The fusion proteins can be specifically and efficiently detected by biotinylated trypsin or biotinylated anhydrotrypsin, as demonstrated by staining of transfected cells, protein blotting, affinity purification, and a mobility shift assay in SDS/polyacrylamide gels.

Damage detection and conductivity evolution in carbon nanofiber epoxy via electrical impedance tomography

International Nuclear Information System (INIS)

Tallman, T N; Wang, K W; Gungor, S; Bakis, C E

2014-01-01

Utilizing electrically conductive nanocomposites for integrated self-sensing and health monitoring is a promising area of structural health monitoring (SHM) research wherein local changes in conductivity coincide with damage. In this research we conduct proof of concept investigations using electrical impedance tomography (EIT) for damage detection by identifying conductivity changes and by imaging conductivity evolution in a carbon nanofiber (CNF) filled epoxy composite. CNF/epoxy is examined because fibrous composites can be manufactured with a CNF/epoxy matrix thereby enabling the entire matrix to become self-sensing. We also study the mechanisms of conductivity evolution in CNF/epoxy through electrical impedance spectroscopy (EIS) testing. The results of these tests indicate that thermal expansion is responsible for conductivity evolution in a CNF/epoxy composite. (paper)

Preterm labour detection by use of a biophysical marker: the uterine electrical activity.

Science.gov (United States)

Marque, Catherine K; Terrien, Jérémy; Rihana, Sandy; Germain, Guy

2007-06-01

The electrical activity of the uterine muscle is representative of uterine contractility. Its characterization may be used to detect a potential risk of preterm delivery in women, even at an early gestational stage. We have investigated the effect of the recording electrode position on the spectral content of the signal by using a mathematical model of the women's abdomen. We have then compared the simulated results to actual recordings. On signals with noise reduced with a dedicated algorithm, we have characterized the main frequency components of the signal spectrum in order to compute parameters indicative of different situations: preterm contractions resulting nonetheless in term delivery (i.e. normal contractions) and preterm contractions leading to preterm delivery (i.e. high-risk contractions). A diagnosis system permitted us to discriminate between these different categories of contractions. As the position of the placenta seems to affect the frequency content of electrical activity, we have also investigated in monkeys, with internal electrodes attached on the uterus, the effect of the placenta on the spectral content of the electrical signals. In women, the best electrode position was the median vertical axis of the abdomen. The discrimination between high risk and normal contractions showed that it was possible to detect a risk of preterm labour as early as at the 27th week of pregnancy (Misclassification Rate range: 11-19.5%). Placental influence on electrical signals was evidenced in animal recordings, with higher energy content in high frequency bands, for signals recorded away from the placenta when compared to signals recorded above the placental insertion. However, we noticed, from pregnancy to labour, a similar evolution of the frequency content of the signal towards high frequencies, whatever the relative position of electrodes and placenta. On human recordings, this study has proved that it is possible to detect, by non-invasive abdominal

Preterm labour detection by use of a biophysical marker: the uterine electrical activity

Directory of Open Access Journals (Sweden)

Germain Guy

2007-06-01

Full Text Available Abstract Background The electrical activity of the uterine muscle is representative of uterine contractility. Its characterization may be used to detect a potential risk of preterm delivery in women, even at an early gestational stage. Methods We have investigated the effect of the recording electrode position on the spectral content of the signal by using a mathematical model of the women's abdomen. We have then compared the simulated results to actual recordings. On signals with noise reduced with a dedicated algorithm, we have characterized the main frequency components of the signal spectrum in order to compute parameters indicative of different situations: preterm contractions resulting nonetheless in term delivery (i.e. normal contractions and preterm contractions leading to preterm delivery (i.e. high-risk contractions. A diagnosis system permitted us to discriminate between these different categories of contractions. As the position of the placenta seems to affect the frequency content of electrical activity, we have also investigated in monkeys, with internal electrodes attached on the uterus, the effect of the placenta on the spectral content of the electrical signals. Results In women, the best electrode position was the median vertical axis of the abdomen. The discrimination between high risk and normal contractions showed that it was possible to detect a risk of preterm labour as early as at the 27th week of pregnancy (Misclassification Rate range: 11–19.5%. Placental influence on electrical signals was evidenced in animal recordings, with higher energy content in high frequency bands, for signals recorded away from the placenta when compared to signals recorded above the placental insertion. However, we noticed, from pregnancy to labour, a similar evolution of the frequency content of the signal towards high frequencies, whatever the relative position of electrodes and placenta. Conclusion On human recordings, this study has

Label-free SnO2 nanowire FET biosensor for protein detection

Science.gov (United States)

Jakob, Markus H.; Dong, Bo; Gutsch, Sebastian; Chatelle, Claire; Krishnaraja, Abinaya; Weber, Wilfried; Zacharias, Margit

2017-06-01

Novel tin oxide field-effect-transistors (SnO2 NW-FET) for pH and protein detection applicable in the healthcare sector are reported. With a SnO2 NW-FET the proof-of-concept of a bio-sensing device is demonstrated using the carrier transport control of the FET channel by a (bio-) liquid modulated gate. Ultra-thin Al2O3 fabricated by a low temperature atomic layer deposition (ALD) process represents a sensitive layer to H+ ions safeguarding the nanowire at the same time. Successful pH sensitivity is demonstrated for pH ranging from 3 to 10. For protein detection, the SnO2 NW-FET is functionalized with a receptor molecule which specifically interacts with the protein of interest to be detected. The feasibility of this approach is demonstrated via the detection of a biotinylated protein using a NW-FET functionalized with streptavidin. An immediate label-free electronic read-out of the signal is shown. The well-established Enzyme-Linked Immunosorbent Assay (ELISA) method is used to determine the optimal experimental procedure which would enable molecular binding events to occur while being compatible with a final label-free electronic read-out on a NW-FET. Integration of the bottom-up fabricated SnO2 NW-FET pH- and biosensor into a microfluidic system (lab-on-a-chip) allows the automated analysis of small volumes in the 400 μl range as would be desired in portable on-site point-of-care (POC) devices for medical diagnosis.

Study of hydrogenated amorphous silicon devices under intense electric field: application to nuclear detection

International Nuclear Information System (INIS)

Ilie, A.

1996-01-01

The goal of this work was the study, development and optimization of hydrogenated amorphous silicon (a-Si:H) devices for use in detection of ionizing radiation in applications connected to the nuclear industry. Thick p-i-n devices, capable of withstanding large electric fields (up to 10 6 V/cm) with small currents (nA/cm 2 ), were proposed and developed. In order to decrease fabrication time, films were made using the 'He diluted' PECVD process and compared to standard a-Si:H films. Aspects connected to specific detector applications as well as to the fundamental physics of a-Si:H were considered: the internal electric field technique, in which the depletion charge was measured as a function of the applied bias voltage; study of the leakage current of p-i-n devices permitted us to demonstrate different regimes: depletion, field-enhanced thermal generation and electronic injection across the p layer. The effect of the electric field on the thermal generation of the carriers was studied considering the Poole-Frenkel and tunneling mechanisms. A model was developed taking under consideration the statistics of the correlated states and electron-phonon coupling. The results suggest that mechanisms not included in the 'standard model' of a Si:h need to be considered, such as defect relaxation, a filed-dependent mobility edge etc...; a new metastable phenomenon, called 'forming', induced by prolonged exposure to a strong electric field, was observed and studied. It is characterized by marked decrease of the leakage current and the detector noise, and increase in the breakdown voltage, as well as an improvement of carrier collection efficiency. This forming process appears to be principally due to an activation of the dopants in the p layer; finally, the capacity of thick p-i-n a Si:H devices to detect ionizing radiation has been evaluated. We show that it is possible, with 20-50 micron thick p-i-n devices, to detect the full spectrum of alpha and beta particles. With an

Automated 3D-Printed Unibody Immunoarray for Chemiluminescence Detection of Cancer Biomarker Proteins

Science.gov (United States)

Tang, C. K.; Vaze, A.; Rusling, J. F.

2017-01-01

A low cost three-dimensional (3D) printed clear plastic microfluidic device was fabricated for fast, low cost automated protein detection. The unibody device features three reagent reservoirs, an efficient 3D network for passive mixing, and an optically transparent detection chamber housing a glass capture antibody array for measuring chemiluminescence output with a CCD camera. Sandwich type assays were built onto the glass arrays using a multi-labeled detection antibody-polyHRP (HRP = horseradish peroxidase). Total assay time was ~30 min in a complete automated assay employing a programmable syringe pump so that the protocol required minimal operator intervention. The device was used for multiplexed detection of prostate cancer biomarker proteins prostate specific antigen (PSA) and platelet factor 4 (PF-4). Detection limits of 0.5 pg mL−1 were achieved for these proteins in diluted serum with log dynamic ranges of four orders of magnitude. Good accuracy vs ELISA was validated by analyzing human serum samples. This prototype device holds good promise for further development as a point-of-care cancer diagnostics tool. PMID:28067370

A simple detection method for low-affinity membrane protein interactions by baculoviral display.

Directory of Open Access Journals (Sweden)

Toshiko Sakihama

Full Text Available BACKGROUND: Membrane protein interactions play an important role in cell-to-cell recognition in various biological activities such as in the immune or neural system. Nevertheless, there has remained the major obstacle of expression of the membrane proteins in their active form. Recently, we and other investigators found that functional membrane proteins express on baculovirus particles (budded virus, BV. In this study, we applied this BV display system to detect interaction between membrane proteins important for cell-to-cell interaction in immune system. METHODOLOGY/PRINCIPAL FINDINGS: We infected Sf9 cells with recombinant baculovirus encoding the T cell membrane protein CD2 or its ligand CD58 and recovered the BV. We detected specific interaction between CD2-displaying BV and CD58-displaying BV by an enzyme-linked immunosorbent assay (ELISA. Using this system, we also detected specific interaction between two other membrane receptor-ligand pairs, CD40-CD40 ligand (CD40L, and glucocorticoid-induced TNFR family-related protein (GITR-GITR ligand (GITRL. Furthermore, we observed specific binding of BV displaying CD58, CD40L, or GITRL to cells naturally expressing their respective receptors by flowcytometric analysis using anti-baculoviral gp64 antibody. Finally we isolated CD2 cDNA from a cDNA expression library by magnetic separation using CD58-displaying BV and anti-gp64 antibody. CONCLUSIONS: We found the BV display system worked effectively in the detection of the interaction of membrane proteins. Since various membrane proteins and their oligomeric complexes can be displayed on BV in the native form, this BV display system should prove highly useful in the search for natural ligands or to develop screening systems for therapeutic antibodies and/or compounds.

Electrophoretic detection of protein p53 in human leukocytes

International Nuclear Information System (INIS)

Paponov, V.D.; Kupsik, E.G.; Shcheglova, E.G.; Yarullin, N.N.

1986-01-01

The authors have found an acid-soluble protein with mol. wt. of about 53 kD in peripheral blood leukocytes of persons with Down's syndrome. It was present in different quantities in all 20 patients tested, but was virtually not discovered in 12 healthy blood donors. This paper determines the possible identity of this protein with protein p53 from mouse ascites carcinoma by comparing their electrophoretic mobilities, because the accuracy of electrophoretic determination of the molecular weight of proteins is not sufficient to identify them. The paper also describes experiments to detect a protein with electrophoretic mobility identical with that of a protein in the leukocytes of patients with Down's syndrome in leukocytes of patients with leukemia. To discover if protein p53 is involved in cell proliferation, the protein composition of leukocytes from healthy blood donors, cultured in the presence and absence of phytohemagglutinin (PHA), was compared. Increased incorporation of H 3-thymidine by leukocytes of patients with Down's syndrome is explained by the presence of a population of immature leukocytes actively synthesizing DNA in the peripheral blood of these patients, and this can also explain the presence of protein p53 in the leukocytes of these patients

Detection of proteins using a colorimetric bio-barcode assay.

Science.gov (United States)

Nam, Jwa-Min; Jang, Kyung-Jin; Groves, Jay T

2007-01-01

The colorimetric bio-barcode assay is a red-to-blue color change-based protein detection method with ultrahigh sensitivity. This assay is based on both the bio-barcode amplification method that allows for detecting miniscule amount of targets with attomolar sensitivity and gold nanoparticle-based colorimetric DNA detection method that allows for a simple and straightforward detection of biomolecules of interest (here we detect interleukin-2, an important biomarker (cytokine) for many immunodeficiency-related diseases and cancers). The protocol is composed of the following steps: (i) conjugation of target capture molecules and barcode DNA strands onto silica microparticles, (ii) target capture with probes, (iii) separation and release of barcode DNA strands from the separated probes, (iv) detection of released barcode DNA using DNA-modified gold nanoparticle probes and (v) red-to-blue color change analysis with a graphic software. Actual target detection and quantification steps with premade probes take approximately 3 h (whole protocol including probe preparations takes approximately 3 days).

Electricity-free, sequential nucleic acid and protein isolation.

Science.gov (United States)

Pawlowski, David R; Karalus, Richard J

2012-05-15

Traditional and emerging pathogens such as Enterohemorrhagic Escherichia coli (EHEC), Yersinia pestis, or prion-based diseases are of significant concern for governments, industries and medical professionals worldwide. For example, EHECs, combined with Shigella, are responsible for the deaths of approximately 325,000 children each year and are particularly prevalent in the developing world where laboratory-based identification, common in the United States, is unavailable (1). The development and distribution of low cost, field-based, point-of-care tools to aid in the rapid identification and/or diagnosis of pathogens or disease markers could dramatically alter disease progression and patient prognosis. We have developed a tool to isolate nucleic acids and proteins from a sample by solid-phase extraction (SPE) without electricity or associated laboratory equipment (2). The isolated macromolecules can be used for diagnosis either in a forward lab or using field-based point-of-care platforms. Importantly, this method provides for the direct comparison of nucleic acid and protein data from an un-split sample, offering a confidence through corroboration of genomic and proteomic analysis. Our isolation tool utilizes the industry standard for solid-phase nucleic acid isolation, the BOOM technology, which isolates nucleic acids from a chaotropic salt solution, usually guanidine isothiocyanate, through binding to silica-based particles or filters (3). CUBRC's proprietary solid-phase extraction chemistry is used to purify protein from chaotropic salt solutions, in this case, from the waste or flow-thru following nucleic acid isolation(4). By packaging well-characterized chemistries into a small, inexpensive and simple platform, we have generated a portable system for nucleic acid and protein extraction that can be performed under a variety of conditions. The isolated nucleic acids are stable and can be transported to a position where power is available for PCR amplification

Method for early detection of infectious mononucleosis by identifying Inmono proteins

Science.gov (United States)

Willard, Karen E.

1984-01-01

Early detection of infectious mononucleosis is carried out using a sample of human blood by isolating and identifying the presence of Inmono proteins in the sample from a two-dimensional protein map with the proteins being characterized by having isoelectric banding as measured in urea of about -16 to -17 with respect to certain isoelectric point standards and molecular mass of about 70 to 75 K daltons as measured in the presence of sodium dodecylsulfate containing polyacrylamide gels, the presence of the Inmono proteins being correlated with the existence of infectious mononucleosis.

Diagnostic value of multi-tumor markers protein biochip detection for primary pulmonary cancer

International Nuclear Information System (INIS)

Xu Fengpo; Wu Yiwei; Li Qingru; Fa Yihua

2005-01-01

To evaluate the diagnostic value of multi-tumor markers protein biochip detection for primary pulmonary cancer, 12 tumor markers including AFP, CEA, NSE, CA125, CA15-3, CA242, CA19-9, PSA, f-PSA, FER, β-HCG and HGH were measured by the protein biochip in the serum of 45 primary pulmonary cancer patients. Positive rate of tumor markers was FER (42.2%), CEA (35.6%), CA125 (24.4%), CA15-3 (17.8%), CA242 (13.3%), CA19-9 (11.1%), β-HCG(8.9%), HGH(6.7%), NSE(4.4%), AFP (0), f-PSA (0) and PSA (0), respectively. The rate of patients with one abnorma indicator was 57.8% except FER. The positive rate using multi-tumor markers protein biochip detection was significantly higher than that of single tumor marker detective method, and this detection can be used for the diagnosis of patients with primary pulmonary cancer. (authors)

Protein sensing by nanofluidic crystal and its signal enhancement

Science.gov (United States)

Sang, Jianming; Du, Hongtan; Wang, Wei; Chu, Ming; Wang, Yuedan; Li, Haichao; Alice Zhang, Haixia; Wu, Wengang; Li, Zhihong

2013-01-01

Nanofluidics has a unique property that ionic conductance across a nanometer-sized confined space is strongly affected by the space surface charge density, which can be utilized to construct electrical read-out biosensor. Based on this principle, this work demonstrated a novel protein sensor along with a sandwich signal enhancement approach. Nanoparticles with designed aptamer onside are assembled in a suspended micropore to form a 3-dimensional network of nanometer-sized interstices, named as nanofluidic crystal hereafter, as the basic sensing unit. Proteins captured by aptamers will change the surface charge density of nanoparticles and thereby can be detected by monitoring the ionic conductance across this nanofluidic crystal. Another aptamer can further enlarge the variations of the surface charge density by forming a sandwich structure (capturing aptamer/protein/signal enhancement aptamer) and the read-out conductance as well. The preliminary experimental results indicated that human α-thrombin was successfully detected by the corresponding aptamer modified nanofluidic crystal with the limit of detection of 5 nM (0.18 μg/ml) and the read-out signal was enhanced up to 3 folds by using another thrombin aptamer. Being easy to graft probe, facile and low-cost to prepare the nano-device, and having an electrical read-out, the present nanofluidic crystal scheme is a promising and universal strategy for protein sensing. PMID:24404017

Electrically detected displacement assay (EDDA): a practical approach to nucleic acid testing in clinical or medical diagnosis.

Science.gov (United States)

Liepold, P; Kratzmüller, T; Persike, N; Bandilla, M; Hinz, M; Wieder, H; Hillebrandt, H; Ferrer, E; Hartwich, G

2008-07-01

This paper introduces the electrically detected displacement assay (EDDA), a electrical biosensor detection principle for applications in medical and clinical diagnosis, and compares the method to currently available microarray technologies in this field. The sensor can be integrated into automated systems of routine diagnosis, but may also be used as a sensor that is directly applied to the polymerase chain reaction (PCR) reaction vessel to detect unlabeled target amplicons within a few minutes. Major aspects of sensor assembly like immobilization procedure, accessibility of the capture probes, and prevention from nonspecific target adsorption, that are a prerequisite for a robust and reliable performance of the sensor, are demonstrated. Additionally, exemplary results from a human papillomavirus assay are presented.

Proteinuria: The diagnostic strategy based on urine proteins differentiation

Directory of Open Access Journals (Sweden)

Stojimirović Biljana B.

2004-01-01

Full Text Available Basal glomerular membrane represents mechanical and electrical barrier for passing of the plasma proteins. Mechanical barrier is composed of cylindrical pores and filtration fissure, and negative layer charge in exterior and interior side of basal glomerular membrane, made of heparan sulphate and sialoglicoproteine, provides certain electrical barrier. Diagnostic strategy based on different serum and urine proteins enables the differentiation of various types of proteinuria. Depending on etiology of proteinuria it can be prerenal, renal and postrenal. By analyzing albumin, armicroglobulin, immunoglobulin G and armacroglobulin, together with total protein in urine, it is possible to detect and differentiate causes of prerenal, renal (glomerular, tubular, glomerulo-tubular and postrenal proteinuria. The adequate and early differentiation of proteinuria type is of an immense diagnostic and therapeutic importance.

Suitability of two-dimensional electrophoretic protein separations for quantitative detection of mutations

International Nuclear Information System (INIS)

Taylor, J.; Anderson, N.L.; Anderson, N.G.; Gemmell, A.; Giometti, C.S.; Nance, S.L.; Tollaksen, S.L.

1986-01-01

Separation of proteins by two-dimensional electrophoresis (2DE) provides a powerful method for mutagenesis studies, since hundreds of proteins can be monitored simultaneously. In previous mutation studies in which 2DE has been used, only qualitative protein differences were monitored; quantitative protein variations were not evaluated. Although significant differences in protein abundance can be detected by eye, the large number of protein spots present in 2DE patterns together with the large number of individual patterns required for a mutagenesis study would necessitate the use of a computerized analysis system to detect the rare quantitative protein changes indicative of gene deletions or inactivation of genes by point mutations in regulatory genes. A pilot study to search for heritable mutations induced by treatment of mice with either ethylnitrosourea or gamma radiation is underway. Samples are being monitored for quantitative changes that reduce the amount of protein by about 50%. The results of this study indicate that the key methods to improve the application of 2DE to mutation screening are to increase the number of measurable spots (i.e., improve stain sensitivity) and to decrease the spread of values for the volume measurements. Even small improvements in these areas could greatly increase the number of monitorable spots. 9 refs., 4 figs

Detection of HOCl-mediated protein oxidation products in the extracellular matrix of human atherosclerotic plaques

DEFF Research Database (Denmark)

Woods, Alan A; Linton, Stuart M; Davies, Michael Jonathan

2003-01-01

Oxidation is believed to play a role in atherosclerosis. Oxidized lipids, sterols and proteins have been detected in early, intermediate and advanced human lesions at elevated levels. The spectrum of oxidized side-chain products detected on proteins from homogenates of advanced human lesions has...... been interpreted in terms of the occurrence of two oxidative mechanisms, one involving oxygen-derived radicals catalysed by trace transition metal ions, and a second involving chlorinating species (HOCl or Cl2), generated by the haem enzyme myeloperoxidase (MPO). As MPO is released extracellularly...... for 83-96% of the total oxidized protein side-chain products detected in these plaques. Oxidation of matrix components extracted from healthy artery tissue, and model proteins, with reagent HOCl is shown to give rise to a similar pattern of products to those detected in advanced human lesions...

Optimization of Xenon Biosensors for Detection of Protein Interactions

International Nuclear Information System (INIS)

Lowery, Thomas J.; Garcia, Sandra; Chavez, Lana; Ruiz, E.Janette; Wu, Tom; Brotin, Thierry; Dutasta, Jean-Pierre; King, David S.; Schultz, Peter G.; Pines, Alex; Wemmer, David E.

2005-08-01

Hyperpolarized 129Xe NMR can detect the presence of specific low-concentration biomolecular analytes by means of the xenon biosensor, which consists of a water-soluble, targeted cryptophane-A cage that encapsulates xenon. In this work we use the prototypical biotinylated xenon biosensor to determine the relationship between the molecular composition of the xenon biosensor and the characteristics of protein-bound resonances. The effects of diastereomer overlap, dipole-dipole coupling, chemical shift anisotropy, xenon exchange, and biosensor conformational exchange on protein-bound biosensor signal were assessed. It was found that optimal protein-bound biosensor signal can be obtained by minimizing the number of biosensor diastereomers and using a flexible linker of appropriate length. Both the linewidth and sensitivity of chemical shift to protein binding of the xenon biosensor were found to be inversely proportional to linker length

Routine detection of calcium-binding proteins following their adsorption to nitrocellulose membrane filters

International Nuclear Information System (INIS)

Hincke, M.T.

1988-01-01

A routine semiquantitative procedure which permits soluble calcium-binding proteins to be detected following their adsorption to nitrocellulose membrane filters by liquid scintillation counting of specifically bound 45 Ca is described. Proteins with high affinity for calcium such as calmodulin and troponin can be detected with a detection threshold of about 2 μg per 400 μl. Modifications to decrease this limit are feasible and are discussed. This technique should allow calcium-binding proteins of unknown function to be assayed during their purification. It was necessary to treat solutions containing 45 Ca with chelex-100 in order to prevent loss of calcium binding which occurred as the decay product (SC 3+ ) accumulated, suggesting that all studies utilizing 45 Ca as a tracer should evaluate possible interference by this ion

Humic substances interfere with detection of pathogenic prion protein

Science.gov (United States)

Smith, Christen B.; Booth, Clarissa J.; Wadzinski, Tyler J.; Legname, Giuseppe; Chappell, Rick; Johnson, Christopher J.; Pedersen, Joel A.

2014-01-01

Studies examining the persistence of prions (the etiological agent of transmissible spongiform encephalopathies) in soil require accurate quantification of pathogenic prion protein (PrPTSE) extracted from or in the presence of soil particles. Here, we demonstrate that natural organic matter (NOM) in soil impacts PrPTSE detection by immunoblotting. Methods commonly used to extract PrPTSE from soils release substantial amounts of NOM, and NOM inhibited PrPTSE immunoblot signal. The degree of immunoblot interference increased with increasing NOM concentration and decreasing NOM polarity. Humic substances affected immunoblot detection of prion protein from both deer and hamsters. We also establish that after interaction with humic acid, PrPTSE remains infectious to hamsters inoculated intracerebrally, and humic acid appeared to slow disease progression. These results provide evidence for interactions between PrPTSE and humic substances that influence both accurate measurement of PrPTSE in soil and disease transmission.

Electrical detection of cellular penetration during microinjection with carbon nanopipettes

Science.gov (United States)

Anderson, Sean E.; Bau, Haim H.

2014-06-01

The carbon nanopipette (CNP) is comprised of a pulled-glass pipette terminating with a nanoscale (tens to hundreds of nm) diameter carbon pipe. The entire inner glass surface of the CNP is coated with a carbon film, providing an electrically conductive path from the carbon tip to the distal, macroscopic end of the pipette. The CNP can double as a nanoelectrode, enabling electrical measurements through its carbon lining, and as a nanoinjector, facilitating reagent injection through its hollow bore. With the aid of a lock-in amplifier, we measured, in real time and with millisecond resolution, variations in impedance and interfacial capacitance as the CNP penetrated into the cytoplasm and nucleus of adherent human osteosarcoma (U20S) cells during microinjection. The capacitance change associated with nucleus penetration was, on average, 1.5 times greater than the one associated with cell membrane penetration. The experimental data was compared and favorably agreed with theoretical predictions based on a simple electrical network model. As a proof of concept, the cytoplasm and nucleus were transfected with fluorescent tRNA, enabling real-time monitoring of tRNA trafficking across the nuclear membrane. The CNP provides a robust and reliable means to detect cell and nucleus penetration, and trigger injection, thereby enabling the automation of cell injection.

Electrical detection of cellular penetration during microinjection with carbon nanopipettes

International Nuclear Information System (INIS)

Anderson, Sean E; Bau, Haim H

2014-01-01

The carbon nanopipette (CNP) is comprised of a pulled-glass pipette terminating with a nanoscale (tens to hundreds of nm) diameter carbon pipe. The entire inner glass surface of the CNP is coated with a carbon film, providing an electrically conductive path from the carbon tip to the distal, macroscopic end of the pipette. The CNP can double as a nanoelectrode, enabling electrical measurements through its carbon lining, and as a nanoinjector, facilitating reagent injection through its hollow bore. With the aid of a lock-in amplifier, we measured, in real time and with millisecond resolution, variations in impedance and interfacial capacitance as the CNP penetrated into the cytoplasm and nucleus of adherent human osteosarcoma (U20S) cells during microinjection. The capacitance change associated with nucleus penetration was, on average, 1.5 times greater than the one associated with cell membrane penetration. The experimental data was compared and favorably agreed with theoretical predictions based on a simple electrical network model. As a proof of concept, the cytoplasm and nucleus were transfected with fluorescent tRNA, enabling real-time monitoring of tRNA trafficking across the nuclear membrane. The CNP provides a robust and reliable means to detect cell and nucleus penetration, and trigger injection, thereby enabling the automation of cell injection. (papers)

An improved method for the immunological detection of mineral bound protein using hydrofluoric acid and direct capture.

Science.gov (United States)

Craig, O E; Collins, M J

2000-03-06

Immunological detection of proteins adsorbed to mineral and ceramic surfaces has proved not only difficult but controversial. Unlike the immunological detection of proteins associated with carbonate or phosphate minerals (e.g. shells and bones) proteins adsorbed to siliceous minerals cannot readily be removed by dissolution of the mineral phase. We have previously examined alternative extraction methodologies which claim to bring the protein into solution, but found none of these to be effective. Here we report a novel strategy for immuno-detection of proteins adsorbed to siliceous minerals, the Digestion and Capture Immunoassay (DACIA). The method involves the use of cold, concentrated (4M) hydrofluoric acid (HF) with the simultaneous capture of liberated protein onto a solid phase. The combination of low temperatures and surface stabilisation enables us to detect epitopes from even partially degraded proteins. The method may have a wide application in forensic, archaeological, soil and earth sciences.

P-aminobenzoic acid and tritiated cyanoborohydride for the detection of pyruvoyl residues in proteins

International Nuclear Information System (INIS)

van Poelje, P.D.; Snell, E.E.

1987-01-01

A procedure for the detection of covalently bound pyruvic acid in purified proteins or in crude extracts is described. The dialyzed sample is first treated with sodium cyanoborohydride to reduce any Schiff bases present and then incubated with p-aminobenzoic acid and sodium [ 3 H]cyanoborohydride. Derivatized proteins are visualized by fluorography following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gel slices containing the labeled proteins are hydrolyzed, and, after removal of polyacrylic acid, the hydrolysate is subjected to ion-exchange high-performance liquid chromatography. The presence of pyruvic acid is established by the detection of a tritiated, 280-nm absorbing compound with a retention time corresponding to that of synthetic N-(p-carboxyphenyl)alanine. The procedure is capable of detecting protein-bound pyruvic acid in the picomolar range and is easily modified to screen for other covalently bound keto acids

Mild electrical stimulation with heat stimulation increase heat shock protein 70 in articular chondrocyte.

Science.gov (United States)

Hiraoka, Nobuyuki; Arai, Yuji; Takahashi, Kenji A; Mazda, Osam; Kishida, Tsunao; Honjo, Kuniaki; Tsuchida, Shinji; Inoue, Hiroaki; Morino, Saori; Suico, Mary Ann; Kai, Hirofumi; Kubo, Toshikazu

2013-06-01

The objective of this study is to investigate the effects of mild electrical stimulation (MES) and heat stress (HS) on heat shock protein 70 (HSP70), that protects chondrocytes and enhances cartilage matrix metabolism, in chondrocyte and articular cartilage. Rabbit articular chondrocytes were treated with MES and/or HS. The safeness was assessed by LDH assay and morphology. HSP70 protein, ubiquitinated proteins and HSP70 mRNA were examined by Western blotting and real-time PCR. Rat knee joints were treated with MES and/or HS. HSP70 protein, ubiquitinated proteins, HSP70 mRNA and proteoglycan core protein (PG) mRNA in articular cartilage were investigated. In vitro, HS increased HSP70 mRNA and HSP70 protein. MES augmented ubiquitinated protein and HSP70 protein, but not HSP70 mRNA. MES + HS raised HSP70 mRNA and ubiquitinated protein, and significantly increased HSP70 protein. In vivo, HS and MES + HS treatment augmented HSP70 mRNA. HS modestly augmented HSP70 protein. MES + HS significantly increased HSP70 protein and ubiquitinated proteins. PG mRNA was markedly raised by MES + HS. This study demonstrated that MES, in combination with HS, increases HSP70 protein in chondrocytes and articular cartilage, and promotes cartilage matrix metabolism in articular cartilage. MES in combination with HS can be a novel physical therapy for osteoarthritis by inducing HSP70 in articular cartilage. Copyright © 2013 Orthopaedic Research Society.

Real-time underwater object detection based on an electrically scanned high-resolution sonar

DEFF Research Database (Denmark)

Henriksen, Lars

1994-01-01

The paper describes an approach to real time detection and tracking of underwater objects, using image sequences from an electrically scanned high-resolution sonar. The use of a high resolution sonar provides a good estimate of the location of the objects, but strains the computers on board, beca...

BioC-compatible full-text passage detection for protein-protein interactions using extended dependency graph.

Science.gov (United States)

Peng, Yifan; Arighi, Cecilia; Wu, Cathy H; Vijay-Shanker, K

2016-01-01

There has been a large growth in the number of biomedical publications that report experimental results. Many of these results concern detection of protein-protein interactions (PPI). In BioCreative V, we participated in the BioC task and developed a PPI system to detect text passages with PPIs in the full-text articles. By adopting the BioC format, the output of the system can be seamlessly added to the biocuration pipeline with little effort required for the system integration. A distinctive feature of our PPI system is that it utilizes extended dependency graph, an intermediate level of representation that attempts to abstract away syntactic variations in text. As a result, we are able to use only a limited set of rules to extract PPI pairs in the sentences, and additional rules to detect additional passages for PPI pairs. For evaluation, we used the 95 articles that were provided for the BioC annotation task. We retrieved the unique PPIs from the BioGRID database for these articles and show that our system achieves a recall of 83.5%. In order to evaluate the detection of passages with PPIs, we further annotated Abstract and Results sections of 20 documents from the dataset and show that an f-value of 80.5% was obtained. To evaluate the generalizability of the system, we also conducted experiments on AIMed, a well-known PPI corpus. We achieved an f-value of 76.1% for sentence detection and an f-value of 64.7% for unique PPI detection.Database URL: http://proteininformationresource.org/iprolink/corpora. © The Author(s) 2016. Published by Oxford University Press.

Nitrile Probes of Electric Field Agree with Independently Measured Fields in Green Fluorescent Protein Even in the Presence of Hydrogen Bonding.

Science.gov (United States)

Slocum, Joshua D; Webb, Lauren J

2016-05-25

There is growing interest in using the nitrile vibrational oscillation as a site-specific probe of local environment to study dynamics, folding, and electrostatics in biological molecules such as proteins. Nitrile probes have been used extensively as reporters of electric field using vibrational Stark effect spectroscopy. However, the analysis of frequencies in terms of electric fields is potentially complicated by the large ground state dipole moment of the nitrile, which may irrevocably perturb the protein under investigation, and the ability of nitriles to accept hydrogen bonds, which causes frequency shifts that are not described by the Stark effect. The consequence of this is that vibrational spectroscopy of nitriles in biomolecules could be predominately sensitive to their local hydration status, not electrostatic environment, and have the potential to be particularly destabilizing to the protein. Here, we introduce green fluorescent protein (GFP) as a model system for addressing these concerns using biosynthetically incorporated p-cyanophenylalanine (pCNF) residues in the interior of GFP and measuring absorption energies of both the intrinsic GFP fluorophore and pCNF residues in response to a series of amino acid mutations. We show that observed changes in emission energy of GFP due to the mutations strongly correlate with changes in electric field experienced by both the nitrile probes and the intrinsic fluorophore. Additionally, we show that changes in electric field measured from the intrinsic fluorophore due to amino acid mutations are unperturbed by the addition of pCNF residues inserted nearby. Finally, we show that changes in electric field experienced by the vibrational probes trend monotonically with changes in field experienced by the native fluorophore even though the nitrile probe is engaged in moderate hydrogen bonding to nearby water molecules, indicated by the temperature dependence of the nitrile's absorption energy. Together these results

Identification of Surface Protein Biomarkers of Listeria monocytogenes via Bioinformatics and Antibody-Based Protein Detection Tools

Science.gov (United States)

Zhang, Cathy X. Y.; Brooks, Brian W.; Huang, Hongsheng; Pagotto, Franco

2016-01-01

ABSTRACT The Gram-positive bacterium Listeria monocytogenes causes a significant percentage of the fatalities among foodborne illnesses in humans. Surface proteins specifically expressed in a wide range of L. monocytogenes serotypes under selective enrichment culture conditions could serve as potential biomarkers for detection and isolation of this pathogen via antibody-based methods. Our study aimed to identify such biomarkers. Interrogation of the L. monocytogenes serotype 4b strain F2365 genome identified 130 putative or known surface proteins. The homologues of four surface proteins, LMOf2365_0578, LMOf2365_0581, LMOf2365_0639, and LMOf2365_2117, were assessed as biomarkers due to the presence of conserved regions among strains of L. monocytogenes which are variable among other Listeria species. Rabbit polyclonal antibodies against the four recombinant proteins revealed the expression of only LMOf2365_0639 on the surface of serotype 4b strain LI0521 cells despite PCR detection of mRNA transcripts for all four proteins in the organism. Three of 35 monoclonal antibodies (MAbs) to LMOf2365_0639, MAbs M3643, M3644, and M3651, specifically recognized 42 (91.3%) of 46 L. monocytogenes lineage I and II isolates grown in nonselective brain heart infusion medium. While M3644 and M3651 reacted with 14 to 15 (82.4 to 88.2%) of 17 L. monocytogenes lineage I and II isolates, M3643 reacted with 22 (91.7%) of 24 lineage I, II, and III isolates grown in selective enrichment media (UVM1, modified Fraser, Palcam, and UVM2 media). The three MAbs exhibited only weak reactivities (the optical densities at 414 nm were close to the cutoff value) to some other Listeria species grown in selective enrichment media. Collectively, the data indicate the potential of LMOf2365_0639 as a surface biomarker of L. monocytogenes, with the aid of specific MAbs, for pathogen detection, identification, and isolation in clinical, environmental, and food samples. IMPORTANCE L. monocytogenes is

Simple fluorescence-based detection of protein kinase A activity using a molecular beacon probe.

Science.gov (United States)

Ma, Changbei; Lv, Xiaoyuan; Wang, Kemin; Jin, Shunxin; Liu, Haisheng; Wu, Kefeng; Zeng, Weimin

2017-11-02

Protein kinase A was detected by quantifying the amount of ATP used after a protein kinase reaction. The ATP assay was performed using the T4 DNA ligase and a molecular beacon (MB). In the presence of ATP, DNA ligase catalyzed the ligation of short DNA. The ligation product then hybridized to MB, resulting in a fluorescence enhancement of the MB. This assay was capable of determining protein kinase A in the range of 12.5∼150 nM, with a detection limit of 1.25 nM. Furthermore, this assay could also be used to investigate the effect of genistein on protein kinase A. It was a universal, non-radioisotopic, and homogeneous method for assaying protein kinase A.

Rapid and label-free detection of protein a by aptamer-tethered porous silicon nanostructures.

Science.gov (United States)

Urmann, Katharina; Reich, Peggy; Walter, Johanna-Gabriela; Beckmann, Dieter; Segal, Ester; Scheper, Thomas

2017-09-10

Protein A, which is secreted by and displayed on the cell membrane of Staphylococcus aureus is an important biomarker for S. aureus. Thus, its rapid and specific detection may facilitate the pathogen identification and initiation of proper treatment. Herein, we present a simple, label-free and rapid optical biosensor enabling specific detection of protein A. Protein A-binding aptamer serves as the capture probe and is immobilized onto a nanostructured porous silicon thin film, which serves as the optical transducer element. We demonstrate high sensitivity of the biosensor with a linear detection range between 8 and 23μM. The apparent dissociation constant was determined as 13.98μM and the LoD is 3.17μM. Harnessing the affinity between protein A and antibodies, a sandwich assay format was developed to amplify the optical signal associated with protein A capture by the aptamer. Using this approach, we increase the sensitivity of the biosensor, resulting in a three times lower LoD. Copyright © 2017 Elsevier B.V. All rights reserved.

Electric field detection of coherent synchrotron radiation in a storage ring generated using laser bunch slicing

International Nuclear Information System (INIS)

Katayama, I.; Shimosato, H.; Bito, M.; Furusawa, K.; Adachi, M.; Zen, H.; Kimura, S.; Katoh, M.; Shimada, M.; Yamamoto, N.; Hosaka, M.; Ashida, M.

2012-01-01

The electric field of coherent synchrotron radiation (CSR) generated by laser bunch slicing in a storage ring has been detected by an electro-optic sampling method. The gate pulses for sampling are sent through a large-mode-area photonic-crystal fiber. The observed electric field profile of the CSR is in good agreement with the spectrum of the CSR observed using Fourier transform far-infrared spectrometry, indicating good phase stability in the CSR. The longitudinal density profiles of electrons modulated by laser pulses were evaluated from the electric field profile.

Fault detection of the connection of lithium-ion power batteries based on entropy for electric vehicles

Science.gov (United States)

Yao, Lei; Wang, Zhenpo; Ma, Jun

2015-10-01

This paper proposes a method of fault detection of the connection of Lithium-Ion batteries based on entropy for electric vehicle. In electric vehicle operation process, some factors, such as road conditions, driving habits, vehicle performance, always affect batteries by vibration, which easily cause loosing or virtual connection between batteries. Through the simulation of the battery charging and discharging experiment under vibration environment, the data of voltage fluctuation can be obtained. Meanwhile, an optimal filtering method is adopted using discrete cosine filter method to analyze the characteristics of system noise, based on the voltage set when batteries are working under different vibration frequency. Experimental data processed by filtering is analyzed based on local Shannon entropy, ensemble Shannon entropy and sample entropy. And the best way to find a method of fault detection of the connection of lithium-ion batteries based on entropy is presented for electric vehicle. The experimental data shows that ensemble Shannon entropy can predict the accurate time and the location of battery connection failure in real time. Besides electric-vehicle industry, this method can also be used in other areas in complex vibration environment.

Direct biosensor immunoassays for the detection of nonmilk proteins in milk powder

NARCIS (Netherlands)

Haasnoot, W.; Olieman, K.; Cazemier, G.; Verheijen, R.

2001-01-01

The low prices of some nonmilk proteins make them attractive as potential adulterants in dairy products. An optical biosensor (BIACORE 3000) was used to develop a direct and combined biosensor immunoassay (BIA) for the simultaneous detection of soy, pea, and soluble wheat proteins in milk powders.

Replacement Condition Detection of Railway Point Machines Using an Electric Current Sensor.

Science.gov (United States)

Sa, Jaewon; Choi, Younchang; Chung, Yongwha; Kim, Hee-Young; Park, Daihee; Yoon, Sukhan

2017-01-29

Detecting replacement conditions of railway point machines is important to simultaneously satisfy the budget-limit and train-safety requirements. In this study, we consider classification of the subtle differences in the aging effect-using electric current shape analysis-for the purpose of replacement condition detection of railway point machines. After analyzing the shapes of after-replacement data and then labeling the shapes of each before-replacement data, we can derive the criteria that can handle the subtle differences between "does-not-need-to-be-replaced" and "needs-to-be-replaced" shapes. On the basis of the experimental results with in-field replacement data, we confirmed that the proposed method could detect the replacement conditions with acceptable accuracy, as well as provide visual interpretability of the criteria used for the time-series classification.

Risk analysis and detection of thrombosis by measurement of electrical resistivity of blood.

Science.gov (United States)

Sapkota, Achyut; Asakura, Yuta; Maruyama, Osamu; Kosaka, Ryo; Yamane, Takashi; Takei, Masahiro

2013-01-01

Monitoring of thrombogenic process is very important in ventricular assistance devices (VADs) used as temporary or permanent measures in patients with advanced heart failure. Currently, there is a lack of a system which can perform a real-time monitoring of thrombogenic activity. Electrical signals vary according to the change in concentration of coagulation factors as well as the distribution of blood cells, and thus have potential to detect the thrombogenic process in an early stage. In the present work, we have made an assessment of an instrumentation system exploiting the electrical properties of blood. The experiments were conducted using bovine blood. Electrical resistance tomography with eight-electrode sensor was used to monitor the spatio-temporal change in electrical resistivity of blood in thrombogenic and non-thrombogenic condition. Under non-thrombogenic condition, the resistivity was uniform across the cross-section and average resistivity monotonically decreased with time before remaining almost flat. In contrary, under thrombogenic condition, there was non-uniform distribution across the cross-section, and average resistivity fluctuated with time.

Electrical Impedance Spectroscopy for Detection of Cells in Suspensions Using Microfluidic Device with Integrated Microneedles

Directory of Open Access Journals (Sweden)

Muhammad Asraf Mansor

2017-02-01

Full Text Available In this study, we introduce novel method of flow cytometry for cell detection based on impedance measurements. The state of the art method for impedance flow cytometry detection utilizes an embedded electrode in the microfluidic to perform measurement of electrical impedance of the presence of cells at the sensing area. Nonetheless, this method requires an expensive and complicated electrode fabrication process. Furthermore, reuse of the fabricated electrode also requires an intensive and tedious cleaning process. Due to that, we present a microfluidic device with integrated microneedles. The two microneedles are placed at the half height of the microchannel for cell detection and electrical measurement. A commercially-available Tungsten needle was utilized for the microneedles. The microneedles are easily removed from the disposable PDMS (Polydimethylsiloxane microchannel and can be reused with a simple cleaning process, such as washing by ultrasonic cleaning. Although this device was low cost, it preserves the core functionality of the sensor, which is capable of detecting passing cells at the sensing area. Therefore, this device is suitable for low-cost medical and food safety screening and testing process in developing countries.

Enhanced detection of single-cell-secreted proteins using a fluorescent immunoassay on the protein-G-terminated glass substrate

Directory of Open Access Journals (Sweden)

Jeong Y

2015-11-01

Full Text Available Yoon Jeong,1,2 Kwan Hong Lee,1,2 Hansoo Park,3 Jonghoon Choi1,2 1Department of Bionano Technology, Graduate School, Hanyang University, Seoul, 2Department of Bionano Engineering, Hanyang University ERICA, Ansan, 3School of Integrative Engineering, Chung-Ang University, Seoul, South Korea Abstract: We present an evaluation of protein-G-terminated glass slides that may contain a suitable substrate for aligning the orientation of antibodies to obtain better binding moiety to the target antigen. The results of the protein-G-terminated slides were compared with those obtained with epoxy-based slides to evaluate signal enhancement for human immunoglobulin G (IgG targets, and an increase in the average fluorescence intensity was observed for the lowest measurable amount of IgG target in the assay using protein-G-terminated slides. Applying this strategy for signal amplification to single-cell assays improves the limits of detection for human IgG protein and cytokines (interleukin-2 and interferon-γ captured from hybridomas. Our data indicate that protein-G-terminated slides have a higher binding capacity for antigens and have better spot-to-spot consistency than that of traditional epoxy-based slides. These properties would be beneficial in the detection of fine amounts of single-cell-secreted proteins, which may provide key insights into cell–cell communication and immune responses. Keywords: microwell array, antibody’s orientation, single cell analysis, secreted cytokine, protein-G-terminated surface

Ultrananocrystalline Diamond Membranes for Detection of High-Mass Proteins

Science.gov (United States)

Kim, H.; Park, J.; Aksamija, Z.; Arbulu, M.; Blick, R. H.

2016-12-01

Mechanical resonators realized on the nanoscale by now offer applications in mass sensing of biomolecules with extraordinary sensitivity. The general idea is that perfect mechanical mass sensors should be of extremely small size to achieve zepto- or yoctogram sensitivity in weighing single molecules similar to a classical scale. However, the small effective size and long response time for weighing biomolecules with a cantilever restricts their usefulness as a high-throughput method. Commercial mass spectrometry (MS), on the other hand, such as electrospray ionization and matrix-assisted laser desorption and ionization (MALDI) time of flight (TOF) and their charge-amplifying detectors are the gold standards to which nanomechanical resonators have to live up to. These two methods rely on the ionization and acceleration of biomolecules and the following ion detection after a mass selection step, such as TOF. The principle we describe here for ion detection is based on the conversion of kinetic energy of the biomolecules into thermal excitation of chemical vapor deposition diamond nanomembranes via phonons followed by phonon-mediated detection via field emission of thermally emitted electrons. We fabricate ultrathin diamond membranes with large lateral dimensions for MALDI TOF MS of high-mass proteins. These diamond membranes are realized by straightforward etching methods based on semiconductor processing. With a minimal thickness of 100 nm and cross sections of up to 400 ×400 μ m2 , the membranes offer extreme aspect ratios. Ion detection is demonstrated in MALDI TOF analysis over a broad range from insulin to albumin. The resulting data in detection show much enhanced resolution as compared to existing detectors, which can offer better sensitivity and overall performance in resolving protein masses.

Design of electric-field assisted surface plasmon resonance system for the detection of heavy metal ions in water

Energy Technology Data Exchange (ETDEWEB)

Kyaw, Htet Htet [Department of Physics, College of Science, Sultan Qaboos University, P. O. Box 36, Al-Khoud 123 (Oman); Boonruang, Sakoolkan, E-mail: sakoolkan.boonruang@nectec.or.th, E-mail: waleed.m@bu.ac.th [Photonics Technology Laboratory, National Electronics and Computer Technology Center (NECTEC), 112 Thailand Science Park, PathumThani 12120 (Thailand); Mohammed, Waleed S., E-mail: sakoolkan.boonruang@nectec.or.th, E-mail: waleed.m@bu.ac.th [Center of Research in Optoelectronics, Communication and Control Systems (BUCROCCS), School of Engineering, Bangkok University, PathumThani 12120 (Thailand); Dutta, Joydeep [Functional Materials Division, School of Information and Communication Technology, KTH Royal Institute of Technology, Isafjordsgatan 22, SE-164 40 Kista, Stockholm (Sweden)

2015-10-15

Surface Plasmon Resonance (SPR) sensors are widely used in diverse applications. For detecting heavy metal ions in water, surface functionalization of the metal surface is typically used to adsorb target molecules, where the ionic concentration is detected via a resonance shift (resonance angle, resonance wavelength or intensity). This paper studies the potential of a possible alternative approach that could eliminate the need of using surface functionalization by the application of an external electric field in the flow channel. The exerted electrical force on the ions pushes them against the surface for enhanced adsorption; hence it is referred to as “Electric-Field assisted SPR system”. High system sensitivity is achieved by monitoring the time dynamics of the signal shift. The ion deposition dynamics are discussed using a derived theoretical model based on ion mobility in water. On the application of an appropriate force, the target ions stack onto the sensor surface depending on the ionic concentration of target solution, ion mass, and flow rate. In the experimental part, a broad detection range of target cadmium ions (Cd{sup 2+}) in water from several parts per million (ppm) down to a few parts per billion (ppb) can be detected.

Out-of-pile experiments with an electrical boiler for Acoustic Boiling Detection

International Nuclear Information System (INIS)

Aberle, J.; Bartholomay, R.; Reimann, G.; Rohrbacher, H.A.; Schleisiek, K.

1978-03-01

This report contains the experimental results of boiling tests obtained during the first testing phase in spring 1977 with an electrically heated 18-rod boiling generator installed in the sodium tank facility (NABEA) of IRE. The layout and performance of the boiling facility together with its instrumentation and criteria of selection of acoustic sensors for the detection of sodium boiling are described and discussed. The report provides information about the thermodynamics, blockage design and thermal conduction within the range of installation of the electric connecting head. The evaluation of the acoustic signals shows that boiling is indicated promptly and with a sufficiently high signal-to-noise ratio both by solid-born sensors and by high temperature microphones placed in the sodium

Plasmonic welded single walled carbon nanotubes on monolayer graphene for sensing target protein

Energy Technology Data Exchange (ETDEWEB)

Kim, Jangheon; Kim, Soohyun [Department of Mechanical Engineering, Korea Advanced Institute of Science and Technology, 373-1 Guseong, Yuseong, Daejeon 305-806 (Korea, Republic of); Kim, Gi Gyu; Jung, Wonsuk, E-mail: wonsuk81@wku.ac.kr [Department of Mechanical and Automotive Engineering, Wonkwang University, Iksan, Jeonbuk 570-749 (Korea, Republic of)

2016-05-16

We developed plasmonic welded single walled carbon nanotubes (SWCNTs) on monolayer graphene as a biosensor to detect target antigen molecules, fc fusion protein without any treatment to generate binder groups for linker and antibody. This plasmonic welding induces atomic networks between SWCNTs as junctions containing carboxylic groups and improves the electrical sensitivity of a SWCNTs and the graphene membrane to detect target protein. We investigated generation of the atomic networks between SWCNTs by field-emission scanning electron microscopy and atomic force microscopy after plasmonic welding process. We compared the intensity ratios of D to G peaks from the Raman spectra and electrical sheet resistance of welded SWCNTs with the results of normal SWCNTs, which decreased from 0.115 to 0.086 and from 10.5 to 4.12, respectively. Additionally, we measured the drain current via source/drain voltage after binding of the antigen to the antibody molecules. This electrical sensitivity of the welded SWCNTs was 1.55 times larger than normal SWCNTs.

Elastin-like polypeptide switches: A design strategy to detect multimeric proteins.

Science.gov (United States)

Dhandhukia, Jugal P; Brill, Dab A; Kouhi, Aida; Pastuszka, Martha K; MacKay, J Andrew

2017-09-01

Elastin-Like Polypeptides (ELPs) reversibly phase separate in response to changes in temperature, pressure, concentration, pH, and ionic species. While powerful triggers, biological microenvironments present a multitude of more specific biological cues, such as antibodies, cytokines, and cell-surface receptors. To develop better biosensors and bioresponsive drug carriers, rational strategies are required to sense and respond to these target proteins. We recently reported that noncovalent association of two ELP fusion proteins to a "chemical inducer of dimerization" small molecule (1.5 kDa) induces phase separation at physiological temperatures. Having detected a small molecule, here we present the first evidence that ELP multimerization can also detect a much larger (60 kDa) protein target. To demonstrate this strategy, ELPs were biotinylated at their amino terminus and mixed with tetrameric streptavidin. At a stoichiometric ratio of [4:1], two to three biotin-ELPs associate with streptavidin into multimeric complexes with an apparent K d of 5 nM. The increased ELP density around a streptavidin core strongly promotes isothermal phase separation, which was tuned to occur at physiological temperature. This phase separation reverses upon saturation with excess streptavidin, which only favors [1:1] complexes. Together, these findings suggest that ELP association with multimeric biomolecules is a viable strategy to deliberately engineer ELPs that respond to multimeric protein substrates. © 2017 The Protein Society.

Replacement Condition Detection of Railway Point Machines Using an Electric Current Sensor

Science.gov (United States)

Sa, Jaewon; Choi, Younchang; Chung, Yongwha; Kim, Hee-Young; Park, Daihee; Yoon, Sukhan

2017-01-01

Detecting replacement conditions of railway point machines is important to simultaneously satisfy the budget-limit and train-safety requirements. In this study, we consider classification of the subtle differences in the aging effect—using electric current shape analysis—for the purpose of replacement condition detection of railway point machines. After analyzing the shapes of after-replacement data and then labeling the shapes of each before-replacement data, we can derive the criteria that can handle the subtle differences between “does-not-need-to-be-replaced” and “needs-to-be-replaced” shapes. On the basis of the experimental results with in-field replacement data, we confirmed that the proposed method could detect the replacement conditions with acceptable accuracy, as well as provide visual interpretability of the criteria used for the time-series classification. PMID:28146057

Effect of the surface film electric resistance on eddy current detectability of surface cracks in Alloy 600 tubes

International Nuclear Information System (INIS)

Saario, T.; Paine, J.P.N.

1995-01-01

The most widely used technique for NDE of steam generator tubing is eddy current. This technique can reliably detect cracks grown in sodium hydroxide environment only at depths greater than 50% through wall. However, cracking caused by thiosulphate solutions have been detected and sized at shallower depths. The disparity has been proposed to be caused by the different electric resistance of the crack wall surface films and corrosion products in the cracks formed in different environments. This work was undertaken to clarify the role of surface film electric resistance on the disparity found in eddy current detectability of surface cracks in alloy 600 tubes. The proposed model explaining the above mentioned disparity is the following. The detectability of tightly closed cracks by the eddy current technique depends on the electric resistance of the surface films of the crack walls. The nature and resistance of the films which form on the crack walls during operation depends on the composition of the solution inside the crack and close to the crack location. During cooling down of the steam generator, because of contraction and loss of internal pressurization, the cracks are rather tightly closed so that exchange of electrolyte and thus changes in the film properties become difficult. As a result, the surface condition prevailing at high temperature is preserved. If the environment is such that the films formed on the crack walls under operating conditions have low electric resistance, eddy current technique will fail to indicate these cracks or will underestimate the size of these cracks. However, if the electric resistance of the films is high, a tightly closed crack will resemble an open crack and will be easily indicated and correctly sized by eddy current technique

Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR

International Nuclear Information System (INIS)

Takeuchi, Koh; Arthanari, Haribabu; Shimada, Ichio; Wagner, Gerhard

2015-01-01

Detection of 15 N in multidimensional NMR experiments of proteins has sparsely been utilized because of the low gyromagnetic ratio (γ) of nitrogen and the presumed low sensitivity of such experiments. Here we show that selecting the TROSY components of proton-attached 15 N nuclei (TROSY 15 N H ) yields high quality spectra in high field magnets (>600 MHz) by taking advantage of the slow 15 N transverse relaxation and compensating for the inherently low 15 N sensitivity. The 15 N TROSY transverse relaxation rates increase modestly with molecular weight but the TROSY gain in peak heights depends strongly on the magnetic field strength. Theoretical simulations predict that the narrowest line width for the TROSY 15 N H component can be obtained at 900 MHz, but sensitivity reaches its maximum around 1.2 GHz. Based on these considerations, a 15 N-detected 2D 1 H– 15 N TROSY-HSQC ( 15 N-detected TROSY-HSQC) experiment was developed and high-quality 2D spectra were recorded at 800 MHz in 2 h for 1 mM maltose-binding protein at 278 K (τ c  ∼ 40 ns). Unlike for 1 H detected TROSY, deuteration is not mandatory to benefit 15 N detected TROSY due to reduced dipolar broadening, which facilitates studies of proteins that cannot be deuterated, especially in cases where production requires eukaryotic expression systems. The option of recording 15 N TROSY of proteins expressed in H 2 O media also alleviates the problem of incomplete amide proton back exchange, which often hampers the detection of amide groups in the core of large molecular weight proteins that are expressed in D 2 O culture media and cannot be refolded for amide back exchange. These results illustrate the potential of 15 N H -detected TROSY experiments as a means to exploit the high resolution offered by high field magnets near and above 1 GHz

Visualization of lipids and proteins at high spatial and temporal resolution via interferometric scattering (iSCAT) microscopy

Czech Academy of Sciences Publication Activity Database

Spindler, S.; Ehrig, J.; König, K.; Nowak, T.; Piliarik, Marek; Stein, H.E.; Taylor, R. W.; Garanger, E.; Lecommandoux, S.; Alves, I. D.; Sandoghdar, V.

2016-01-01

Roč. 49, č. 27 (2016), č. článku 274002. ISSN 0022-3727 Institutional support: RVO:67985882 Keywords : lipid diffusion * protein detection * single molecule detection Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 2.588, year: 2016

Online fouling detection in electrical circulation heaters using neural networks

Energy Technology Data Exchange (ETDEWEB)

Lalot, S. [M.E.T.I.E.R., Longuenesse Cedex (France); Universite de Valenciennes (France). LME; Lecoeuche, S. [M.E.T.I.E.R., Longuenesse Cedex (France); Universite de Lille (France). Laboratoire 13D

2003-06-01

Here is presented a method that is able to detect fouling during the service of a circulation electrical heater. The neural based technique is divided in two major steps: identification and classification. Each step uses a neural network, the connection weights of the first one being the inputs of the second network. Each step is detailed and the main characteristics and abilities of the two neural networks are given. It is shown that the method is able to discriminate fouling from viscosity modification that would lead to the same type of effect on the total heat transfer coefficient. (author)

Electrical detection of spin transport in lateral ferromagnet-semiconductor devices

Science.gov (United States)

Lou, Xiaohua

2007-03-01

A fully electrical scheme of spin injection, transport, and detection in a single ferromagnet-semiconductor structure has been a long-standing goal in the field of spintronics. In this talk, we report on an experimental demonstration of such a scheme. The devices are fabricated from epitaxial Fe/GaAs (100) heterostructures with highly doped GaAs as a Schottky tunnel barrier. A set of closely spaced Fe contacts on the top of an n-GaAs channel are used as spin injectors and detectors. Reference electrodes are placed at the far ends of the channel, allowing for non-local spin detection [1]. The electro-chemical potential of the detector is sensitive to the relative magnetizations of the injector and detector. In spin-valve measurements, a magnetic field is applied along the Fe easy axis to switch the relative magnetizations of injector and detector from parallel to antiparallel, resulting in a voltage jump that is proportional to the non-equilibrium spin polarization in the channel. A more rigorous test of electrical spin detection is the observation of the Hanle effect, in which an out-of-plane magnetic field is used to modulate and dephase the spin polarization in the channel. The magnitudes of the observed Hanle curves agree with the results of the spin-valve measurements. The dependence of the Hanle curves on temperature and contact separation is studied in detail and is consistent with a drift-diffusion model incorporating spin precession and relaxation. The spin polarization generated by spin injection (reverse bias at the injector) or spin accumulation (forward bias at the injector) is measured using the magneto-optical Kerr effect and is found to be in good agreement with the spin-dependent non-local voltage. Both the transport and optical measurements show a non-linear relationship between the bias voltage at the injector and the spin polarization in the channel. [1] M. Johnson and R. H. Silsbee, Phys. Rev. Lett. 55, 1790 (1985).

Evaluation of back scatter interferometry, a method for detecting protein binding in solution

DEFF Research Database (Denmark)

Jepsen, S. T.; Jørgensen, Thomas Martini; Zong, Weiyong

2015-01-01

Back Scatter Interferometry (BSI) has been proposed to be a highly sensitive and versatile refractive index sensor usable for analytical detection of biomarker and protein interactions in solution. However the existing literature on BSI lacks a physical explanation of why protein interactions in ...

Label and Label-Free Detection Techniques for Protein Microarrays

Directory of Open Access Journals (Sweden)

Amir Syahir

2015-04-01

Full Text Available Protein microarray technology has gone through numerous innovative developments in recent decades. In this review, we focus on the development of protein detection methods embedded in the technology. Early microarrays utilized useful chromophores and versatile biochemical techniques dominated by high-throughput illumination. Recently, the realization of label-free techniques has been greatly advanced by the combination of knowledge in material sciences, computational design and nanofabrication. These rapidly advancing techniques aim to provide data without the intervention of label molecules. Here, we present a brief overview of this remarkable innovation from the perspectives of label and label-free techniques in transducing nano‑biological events.

Phage-Mediated Immuno-PCR for Ultrasensitive Detection of Cry1Ac Protein Based on Nanobody.

Science.gov (United States)

Liu, Yuanyuan; Jiang, Dongjian; Lu, Xin; Wang, Wei; Xu, Yang; He, Qinghua

2016-10-11

The widespread use of Cry proteins in transgenic plants for insect control has raised concerns about the environment and food safety in the public. An effective detection method for introduced Cry proteins is of significance for environmental risk assessment and product quality control. This paper describes a novel phage mediated immuno-PCR (iPCR) for the ultrasensitive determination of Cry proteins based on nanobodies. Three nanobodies against Cry1Ac protein were obtained from a naı̈ve phage displayed nanobody library without animal immunization process and were applied to the iPCR assay for Cry1Ac. The phage-mediated iPCR for Cry1Ac based on nanobodies showed a dynamic range of 0.001-100 ng/mL and a limit detection of 0.1 pg/mL. Specific measurement of this established method was performed by testing cross-reativity of other Cry1Ac analogues, and the result showed negligible cross-reactivity with other test Cry proteins (Cry1Ab, Cry1F, Cry3B). Furthermore, the phage-mediated iPCR based on nanobody should be easily applicable to the detection of many other Cry proteins.

Geo electrical Resistivity Survey for Ancient Tunnel Detection at Bukit Tenggek, Setiu, Terengganu

International Nuclear Information System (INIS)

Siti Nazira Masrom; Mohd Hariri Arifin; Abd Rahim Harun; Abdul Rahim Samsudin

2011-01-01

Geo electrical resistivity survey was conducted in the Bukit Tenggek, Setiu, Terengganu to detect the possible existence of an ancient tunnel which is believed to be in the area. Geo electrical resistivity method was found very effective in searching for archaeological exploration and underground structures (tunnels and artifacts). Geo electrical resistivity survey was carried out using Terrameter ABEM SAS1000 and Wenner array electrode configuration. The survey area is located in a damp valley with a stream across the region. 2-D resistivity image showed the existence of anomalies in several areas that can be associated with the structure. Low resistivity value represents the estimated existence of the old tunnel, while isolated rounded anomalies are believed to be associated with barrels/artifacts. 3-D resistivity profiles, shows anomalies that may be caused by the existence of a horizontal and two vertical tunnels (shaft). However, the drillings work need to be done to figure out the exact cause of these anomalies. (author)

Nuclear magnetic resonance detection and spectroscopy of single proteins using quantum logic.

Science.gov (United States)

Lovchinsky, I; Sushkov, A O; Urbach, E; de Leon, N P; Choi, S; De Greve, K; Evans, R; Gertner, R; Bersin, E; Müller, C; McGuinness, L; Jelezko, F; Walsworth, R L; Park, H; Lukin, M D

2016-02-19

Nuclear magnetic resonance spectroscopy is a powerful tool for the structural analysis of organic compounds and biomolecules but typically requires macroscopic sample quantities. We use a sensor, which consists of two quantum bits corresponding to an electronic spin and an ancillary nuclear spin, to demonstrate room temperature magnetic resonance detection and spectroscopy of multiple nuclear species within individual ubiquitin proteins attached to the diamond surface. Using quantum logic to improve readout fidelity and a surface-treatment technique to extend the spin coherence time of shallow nitrogen-vacancy centers, we demonstrate magnetic field sensitivity sufficient to detect individual proton spins within 1 second of integration. This gain in sensitivity enables high-confidence detection of individual proteins and allows us to observe spectral features that reveal information about their chemical composition. Copyright © 2016, American Association for the Advancement of Science.

Enhanced detection method for corneal protein identification using shotgun proteomics

Directory of Open Access Journals (Sweden)

Schlager John J

2009-06-01

Full Text Available Abstract Background The cornea is a specialized transparent connective tissue responsible for the majority of light refraction and image focus for the retina. There are three main layers of the cornea: the epithelium that is exposed and acts as a protective barrier for the eye, the center stroma consisting of parallel collagen fibrils that refract light, and the endothelium that is responsible for hydration of the cornea from the aqueous humor. Normal cornea is an immunologically privileged tissue devoid of blood vessels, but injury can produce a loss of these conditions causing invasion of other processes that degrade the homeostatic properties resulting in a decrease in the amount of light refracted onto the retina. Determining a measure and drift of phenotypic cornea state from normal to an injured or diseased state requires knowledge of the existing protein signature within the tissue. In the study of corneal proteins, proteomics procedures have typically involved the pulverization of the entire cornea prior to analysis. Separation of the epithelium and endothelium from the core stroma and performing separate shotgun proteomics using liquid chromatography/mass spectrometry results in identification of many more proteins than previously employed methods using complete pulverized cornea. Results Rabbit corneas were purchased, the epithelium and endothelium regions were removed, proteins processed and separately analyzed using liquid chromatography/mass spectrometry. Proteins identified from separate layers were compared against results from complete corneal samples. Protein digests were separated using a six hour liquid chromatographic gradient and ion-trap mass spectrometry used for detection of eluted peptide fractions. The SEQUEST database search results were filtered to allow only proteins with match probabilities of equal or better than 10-3 and peptides with a probability of 10-2 or less with at least two unique peptides isolated within

Manufacturing and testing flexible microfluidic devices with optical and electrical detection mechanisms

Science.gov (United States)

Ivan, Marius G.; Vivet, Frédéric; Meinders, Erwin R.

2010-06-01

Flexible microfluidic devices made of poly(dimethylsiloxane) (PDMS) were manufactured by soft lithography, and tested in detection of ionic species using optical absorption spectroscopy and electrical measurements. PDMS was chosen due to its flexibility and ease of surface modification by exposure to plasma and UV treatment, its transparency in UV-Vis regions of the light spectrum, and biocompatibility. The dual-detection mechanism allows the user more freedom in choosing the detection tool, and a functional device was successfully tested. Optical lithography was employed for manufacturing templates, which were subsequently used for imprinting liquid PDMS by thermal curing. Gold electrodes having various widths and distances among them were patterned with optical lithography on the top part which sealed the microchannels, and the devices were employed for detection of ionic species in aqueous salt solutions as well as micro-electrolysis cells. Due to the transparency of PDMS in UV-Vis the microfluidics were also used as photoreactors, and the in-situ formed charged species were monitored by applying a voltage between electrodes. Upon addition of a colorimetric pH sensor, acid was detected with absorption spectroscopy.

Development of an enzyme-linked immunosorbent assay method to detect mustard protein in mustard seed oil

NARCIS (Netherlands)

Koppelman, S.J.; Vlooswijk, R.; Bottger, G.; Duijn, G. van; Schaft, P. van der; Dekker, J.; Bemgen, H. van

2007-01-01

An enzyme-linked immunosorbent assay for the detection of mustard protein was developed. The assay is based on a polyclonal antiserum directed against a mixture of mustard proteins raised in rabbits. The assay has a detection limit of 1.5 ppm (milligrams per kilogram) and is suitable for the

Oil leakage detection for electric power equipment based on ultraviolet fluorescence effect

Science.gov (United States)

Zhang, Jing; Wang, Jian-hui; Xu, Bin; Huang, Zhi-dong; Huang, Lan-tao

2018-03-01

This paper presents a method to detect the oil leakage of high voltage power equipment based on ultraviolet fluorescence effect. The method exploits the principle that the insulating oil has the fluorescent effect under the irradiation of specific ultraviolet light. The emission spectrum of insulating oil under excitation light with different wavelengths is measured and analyzed first. On this basis, a portable oil leakage detective device for high voltage power equipment is designed and developed with a selected 365 nm ultraviolet as the excitation light and the low light level camera as the fluorescence image collector. Then, the feasibility of the proposed method and device in different conditions is experimentally verified in the laboratory environment. Finally, the developed oil leakage detective device is applied to 500 kV Xiamen substation and Quanzhou substation. And the results show that the device can detect the oil leakage of high voltage electrical equipment quickly and conveniently even under the condition of a slight oil leakage especially in the low light environment.

A Two-Week Guided Inquiry Protein Separation and Detection Experiment for Undergraduate Biochemistry

Science.gov (United States)

Carolan, James P.; Nolta, Kathleen V.

2016-01-01

A laboratory experiment for teaching protein separation and detection in an undergraduate biochemistry laboratory course is described. This experiment, performed in two, 4 h laboratory periods, incorporates guided inquiry principles to introduce students to the concepts behind and difficulties of protein purification. After using size-exclusion…

A Rapid, Onsite, Ultrasensitive Melamine Quantitation Method for Protein Beverages Using Time-Resolved Fluorescence Detection Paper.

Science.gov (United States)

Li, Guanghua; Wang, Du; Zhou, Aijun; Sun, Yimin; Zhang, Qi; Poapolathep, Amnart; Zhang, Li; Fan, Zhiyong; Zhang, Zhaowei; Li, Peiwu

2018-05-02

To ensure protein beverage safety and prevent illegal melamine use to artificially increase protein content, a rapid, onsite, ultrasensitive detection method for melamine must be developed because melamine is detrimental to human health and life. Herein, an ultrasensitive time-resolved fluorescence detection paper (TFDP) was developed to detect melamine in protein beverages within 15 min using a one-step sample preparation. The lower limits of detection were 0.89, 0.94, and 1.05 ng/mL, and the linear ranges were 2.67-150, 2.82-150, and 3.15-150 ng/mL (R2>0.982) for peanut, walnut, and coconut beverages, respectively. The recovery rates were 85.86-110.60% with a coefficient of variation beverage samples, the TFDP and ultra-performance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) results were consistent. This method is a promising alternative for rapid, onsite detection of melamine in beverages.

Progress in proton-detected solid-state NMR (SSNMR): Super-fast 2D SSNMR collection for nano-mole-scale proteins

Science.gov (United States)

Ishii, Yoshitaka; Wickramasinghe, Ayesha; Matsuda, Isamu; Endo, Yuki; Ishii, Yuji; Nishiyama, Yusuke; Nemoto, Takahiro; Kamihara, Takayuki

2018-01-01

Proton-detected solid-state NMR (SSNMR) spectroscopy has attracted much attention due to its excellent sensitivity and effectiveness in the analysis of trace amounts of amyloid proteins and other important biological systems. In this perspective article, we present the recent sensitivity limit of 1H-detected SSNMR using "ultra-fast" magic-angle spinning (MAS) at a spinning rate (νR) of 80-100 kHz. It was demonstrated that the high sensitivity of 1H-detected SSNMR at νR of 100 kHz and fast recycling using the paramagnetic-assisted condensed data collection (PACC) approach permitted "super-fast" collection of 1H-detected 2D protein SSNMR. A 1H-detected 2D 1H-15N correlation SSNMR spectrum for ∼27 nmol of a uniformly 13C- and 15N-labeled GB1 protein sample in microcrystalline form was acquired in only 9 s with 50% non-uniform sampling and short recycle delays of 100 ms. Additional data suggests that it is now feasible to detect as little as 1 nmol of the protein in 5.9 h by 1H-detected 2D 1H-15N SSNMR at a nominal signal-to-noise ratio of five. The demonstrated sensitivity is comparable to that of modern solution protein NMR. Moreover, this article summarizes the influence of ultra-fast MAS and 1H-detection on the spectral resolution and sensitivity of protein SSNMR. Recent progress in signal assignment and structural elucidation by 1H-detected protein SSNMR is outlined with both theoretical and experimental aspects.

Quantum dot bio-conjugate: as a western blot probe for highly sensitive detection of cellular proteins

Energy Technology Data Exchange (ETDEWEB)

Kale, Sonia [Agharkar Research Institute (India); Kale, Anup [University of Alabama, Center for Materials for Information Technology (United States); Gholap, Haribhau; Rana, Abhimanyu [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Desai, Rama [National Centre for Cell Science (India); Banpurkar, Arun [University of Pune, Department of Physics (India); Ogale, Satishchandra, E-mail: sb.ogale@ncl.res.in [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Shastry, Padma, E-mail: padma@nccs.res.in [National Centre for Cell Science (India)

2012-03-15

In the present study, we report a quantum dot (QD)-tailored western blot analysis for a sensitive, rapid and flexible detection of the nuclear and cytoplasmic proteins. Highly luminescent CdTe and (CdTe)ZnS QDs are synthesized by aqueous method. High resolution transmission electron microscopy, Raman spectroscopy, fourier transform infrared spectroscopy, fluorescence spectroscopy and X-ray diffraction are used to characterize the properties of the quantum dots. The QDs are functionalized with antibodies of prostate apoptosis response-4 (Par-4), poly(ADP-ribose) polymerases and {beta} actin to specifically bind with the proteins localized in the nucleus and cytoplasm of the cells, respectively. The QD-conjugated antibodies are used to overcome the limitations of conventional western blot technique. The sensitivity and rapidity of protein detection in QD-based approach is very high, with detection limits up to 10 pg of protein. In addition, these labels provide the capability of enhanced identification and localization of marker proteins in intact cells by confocal laser scanning microscopy.

Detection of needle to nerve contact based on electric bioimpedance and machine learning methods.

Science.gov (United States)

Kalvoy, Havard; Tronstad, Christian; Ullensvang, Kyrre; Steinfeldt, Thorsten; Sauter, Axel R

2017-07-01

In an ongoing project for electrical impedance-based needle guidance we have previously showed in an animal model that intraneural needle positions can be detected with bioimpedance measurement. To enhance the power of this method we in this study have investigated whether an early detection of the needle only touching the nerve also is feasible. Measurement of complex impedance during needle to nerve contact was compared with needle positions in surrounding tissues in a volunteer study on 32 subjects. Classification analysis using Support-Vector Machines demonstrated that discrimination is possible, but that the sensitivity and specificity for the nerve touch algorithm not is at the same level of performance as for intra-neuralintraneural detection.

DETECTION AND PURIFICATION OF TYROSINE-SULFATED PROTEINS USING A NOVEL ANTI-SULFOTYROSINE MONOCLONAL ANTIBODY*

Science.gov (United States)

Hoffhines, Adam J.; Damoc, Eugen; Bridges, Kristie G.; Leary, Julie A.; Moore, Kevin L.

2006-01-01

Protein-tyrosine O-sulfation is a post-translational modification mediated by one of two Golgi tyrosylprotein sulfotransferases (TPST-1 & TPST-2) that catalyze the transfer of sulfate to tyrosine residues in secreted and transmembrane proteins. Tyrosine sulfation plays a role in protein-protein interactions in several well-defined systems. Although dozens of tyrosine-sulfated proteins are known, many more are likely to exist and await description. Advancing our understanding of the importance of tyrosine sulfation in biological systems requires the development of new tools for the detection and study of tyrosine-sulfated proteins. We have developed a novel anti-sulfotyrosine monoclonal antibody, called PSG2, that binds with high affinity and exquisite specificity to sulfotyrosine residues in peptides and proteins independent of sequence context. We show that it can detect tyrosinesulfated proteins in complex biological samples and can be used as a probe to assess the role of tyrosine sulfation in protein function. We also demonstrate the utility of PSG2 in the purification of tyrosine-sulfated proteins from crude tissue samples. Finally, Western blot analysis using PSG2 indicates that certain sperm/epididymal proteins are undersulfated in Tpst2−/− mice. This indicates that TPST-1 and TPST-2 have distinct macromolecular substrate specificities and provides clues as to the molecular mechanism of the infertility of Tpst2−/− males. PSG2 should be widely applicable for identification of tyrosine-sulfated proteins in other systems and organisms. PMID:17046811

Detecting protein-protein interactions in living cells

DEFF Research Database (Denmark)

Gottschalk, Marie; Bach, Anders; Hansen, Jakob Lerche

2009-01-01

to the endogenous C-terminal peptide of the NMDA receptor, as evaluated by a cell-free protein-protein interaction assay. However, it is important to address both membrane permeability and effect in living cells. Therefore a bioluminescence resonance energy transfer (BRET) assay was established, where the C......-terminal of the NMDA receptor and PDZ2 of PSD-95 were fused to green fluorescent protein (GFP) and Renilla luciferase (Rluc) and expressed in COS7 cells. A robust and specific BRET signal was obtained by expression of the appropriate partner proteins and subsequently, the assay was used to evaluate a Tat......The PDZ domain mediated interaction between the NMDA receptor and its intracellular scaffolding protein, PSD-95, is a potential target for treatment of ischemic brain diseases. We have recently developed a number of peptide analogues with improved affinity for the PDZ domains of PSD-95 compared...

Nitrogen detected TROSY at high field yields high resolution and sensitivity for protein NMR

Energy Technology Data Exchange (ETDEWEB)

Takeuchi, Koh [National Institute for Advanced Industrial Science and Technology, Molecular Profiling Research Center for Drug Discovery (Japan); Arthanari, Haribabu [Harvard Medical School, Department of Biochemistry and Molecular Pharmacology (United States); Shimada, Ichio, E-mail: shimada@iw-nmr.f.u-tokyo.ac.jp [National Institute for Advanced Industrial Science and Technology, Molecular Profiling Research Center for Drug Discovery (Japan); Wagner, Gerhard, E-mail: gerhard-wagner@hms.harvard.edu [Harvard Medical School, Department of Biochemistry and Molecular Pharmacology (United States)

2015-12-15

Detection of {sup 15}N in multidimensional NMR experiments of proteins has sparsely been utilized because of the low gyromagnetic ratio (γ) of nitrogen and the presumed low sensitivity of such experiments. Here we show that selecting the TROSY components of proton-attached {sup 15}N nuclei (TROSY {sup 15}N{sub H}) yields high quality spectra in high field magnets (>600 MHz) by taking advantage of the slow {sup 15}N transverse relaxation and compensating for the inherently low {sup 15}N sensitivity. The {sup 15}N TROSY transverse relaxation rates increase modestly with molecular weight but the TROSY gain in peak heights depends strongly on the magnetic field strength. Theoretical simulations predict that the narrowest line width for the TROSY {sup 15}N{sub H} component can be obtained at 900 MHz, but sensitivity reaches its maximum around 1.2 GHz. Based on these considerations, a {sup 15}N-detected 2D {sup 1}H–{sup 15}N TROSY-HSQC ({sup 15}N-detected TROSY-HSQC) experiment was developed and high-quality 2D spectra were recorded at 800 MHz in 2 h for 1 mM maltose-binding protein at 278 K (τ{sub c} ∼ 40 ns). Unlike for {sup 1}H detected TROSY, deuteration is not mandatory to benefit {sup 15}N detected TROSY due to reduced dipolar broadening, which facilitates studies of proteins that cannot be deuterated, especially in cases where production requires eukaryotic expression systems. The option of recording {sup 15}N TROSY of proteins expressed in H{sub 2}O media also alleviates the problem of incomplete amide proton back exchange, which often hampers the detection of amide groups in the core of large molecular weight proteins that are expressed in D{sub 2}O culture media and cannot be refolded for amide back exchange. These results illustrate the potential of {sup 15}N{sub H}-detected TROSY experiments as a means to exploit the high resolution offered by high field magnets near and above 1 GHz.

Electrical Detection of Spin-to-Charge Conversion in a Topological Insulator Bi2Te3

Science.gov (United States)

Li, Connie H.; van't Erve, Olaf M. J.; Li, Yaoyi; Li, Lian; Jonker, Berry T.

Spin-momentum locking in topological insulators (TIs) dictates that an unpolarized charge current creates a net spin polarization. We recently demonstrated the first electrical detection of this spontaneous polarization in a transport geometry, using a ferromagnetic (FM) / tunnel barrier contact, where the projection of the TI surface state spin on the magnetization of detector is measured as a voltage [1]. Alternatively, if spins are injected into the TI surface state system, it is distinctively associated with a unique carrier momentum, and hence should generated a charge accumulation, similar to that of inverse spin Hall effect. Here we experimentally demonstrate both effects in the same device fabricated in Bi2Te3: the electrical detection of the spin accumulation generated by an unpolarized current flowing through the surface states, and that of the charge accumulation generated by spins injected into the surface states system. This reverse measurement is an independent confirmation of spin-momentum locking in the TI surface states, and offers additional avenue for spin manipulation. It further demonstrates the robustness and versatility of electrical access to the TI surface state spin system, an important step towards its utilization in TI-based spintronics devices. C.H. Li et al., Nat. Nanotech. 9, 218 (2014). Supported by NRL core funds and Nanoscience Institute.

Ultrasensitive Detection of Proteins on Western Blots with Semiconducting Polymer Dots

OpenAIRE

Ye, Fangmao; Smith, Polina B.; Wu, Changfeng; Chiu, Daniel T.

2013-01-01

We demonstrate ultrasensitive fluorescence imaging of proteins on Western blots using a bright, compact, and orange-emitting semiconducting polymer dot (CN-PPV). We achieved a detection limit at the single-picogram level in dot blots; with conventional Western blotting, we detected 50 pg of transferrin and trypsin inhibitor after SDS-PAGE and transfer onto a PVDF membrane. Our method does not require any additional equipment or time compared to the conventional procedure with traditional fluo...

Food Forensics: Using Mass Spectrometry To Detect Foodborne Protein Contaminants, as Exemplified by Shiga Toxin Variants and Prion Strains.

Science.gov (United States)

Silva, Christopher J

2018-06-13

Food forensicists need a variety of tools to detect the many possible food contaminants. As a result of its analytical flexibility, mass spectrometry is one of those tools. Use of the multiple reaction monitoring (MRM) method expands its use to quantitation as well as detection of infectious proteins (prions) and protein toxins, such as Shiga toxins. The sample processing steps inactivate prions and Shiga toxins; the proteins are digested with proteases to yield peptides suitable for MRM-based analysis. Prions are detected by their distinct physicochemical properties and differential covalent modification. Shiga toxin analysis is based on detecting peptides derived from the five identical binding B subunits comprising the toxin. 15 N-labeled internal standards are prepared from cloned proteins. These examples illustrate the power of MRM, in that the same instrument can be used to safely detect and quantitate protein toxins, prions, and small molecules that might contaminate our food.

Surface plasmon resonance biosensor for parallelized detection of protein biomarkers in diluted blood plasma

Czech Academy of Sciences Publication Activity Database

Piliarik, Marek; Bocková, Markéta; Homola, Jiří

2010-01-01

Roč. 26, č. 4 (2010), s. 1656-1661 ISSN 0956-5663 R&D Projects: GA AV ČR KAN200670701 Institutional research plan: CEZ:AV0Z20670512 Keywords : Surface plasmon resonance * Protein array * Cancer marker Subject RIV: JA - Electronics ; Optoelectronics, Electrical Engineering Impact factor: 5.361, year: 2010

Nanotechnology for Early Cancer Detection

Directory of Open Access Journals (Sweden)

Joon Won Park

2010-01-01

Full Text Available Vast numbers of studies and developments in the nanotechnology area have been conducted and many nanomaterials have been utilized to detect cancers at early stages. Nanomaterials have unique physical, optical and electrical properties that have proven to be very useful in sensing. Quantum dots, gold nanoparticles, magnetic nanoparticles, carbon nanotubes, gold nanowires and many other materials have been developed over the years, alongside the discovery of a wide range of biomarkers to lower the detection limit of cancer biomarkers. Proteins, antibody fragments, DNA fragments, and RNA fragments are the base of cancer biomarkers and have been used as targets in cancer detection and monitoring. It is highly anticipated that in the near future, we might be able to detect cancer at a very early stage, providing a much higher chance of treatment.

Ageing of insulation and diagnosis of electrical equipment through detection of partial discharge

International Nuclear Information System (INIS)

Lopez Vergara, T.; Velasco Bernal, C.

1994-01-01

Ageing in electrical equipment affects mainly its insulation system. Such ageing in the insulation system is determined by its organic nature, basically constituted by three families of materials: cellulose, resin and hydrocarbon. All of these are affected by high temperatures, which tend to produce a break in the molecular chains (if the temperatures are not too high) or carbonization and gasification of the material (if they are). The radiation absorbed by the insulating materials also destroys molecular chains, causing degradation of the material. The break of the molecular chains, especially in the polymer-based materials, fragments the material, mainly in areas subjected to mechanical forces and stresses. From the electrical point of view, fissures occurring the insulating material lead to a much lower dielectric strength in certain parts of the materials, which could produce partial discharge conditions. Therefore, the growth of partial discharges in electrical equipment items is frequently the consequences of ageing, and be used to evaluate their residual life. Empresarios Agrupados has developed a system to detect partial discharges which can be used while equipment is still in operation. The measurements taken with this system are sufficiently accurate and repetitive to be used in evaluating the condition of medium-voltage electrical equipment insulation. (Author)

Optimizing Urine Processing Protocols for Protein and Metabolite Detection.

Science.gov (United States)

Siddiqui, Nazema Y; DuBois, Laura G; St John-Williams, Lisa; Will, Thompson J; Grenier, Carole; Burke, Emily; Fraser, Matthew O; Amundsen, Cindy L; Murphy, Susan K

In urine, factors such as timing of voids, and duration at room temperature (RT) may affect the quality of recovered protein and metabolite data. Additives may aid with detection, but can add more complexity in sample collection or analysis. We aimed to identify the optimal urine processing protocol for clinically-obtained urine samples that allows for the highest protein and metabolite yields with minimal degradation. Healthy women provided multiple urine samples during the same day. Women collected their first morning (1 st AM) void and another "random void". Random voids were aliquotted with: 1) no additive; 2) boric acid (BA); 3) protease inhibitor (PI); or 4) both BA + PI. Of these aliquots, some were immediately stored at 4°C, and some were left at RT for 4 hours. Proteins and individual metabolites were quantified, normalized to creatinine concentrations, and compared across processing conditions. Sample pools corresponding to each processing condition were analyzed using mass spectrometry to assess protein degradation. Ten Caucasian women between 35-65 years of age provided paired 1 st morning and random voided urine samples. Normalized protein concentrations were slightly higher in 1 st AM compared to random "spot" voids. The addition of BA did not significantly change proteins, while PI significantly improved normalized protein concentrations, regardless of whether samples were immediately cooled or left at RT for 4 hours. In pooled samples, there were minimal differences in protein degradation under the various conditions we tested. In metabolite analyses, there were significant differences in individual amino acids based on the timing of the void. For comparative translational research using urine, information about void timing should be collected and standardized. For urine samples processed in the same day, BA does not appear to be necessary while the addition of PI enhances protein yields, regardless of 4°C or RT storage temperature.

The rational development of molecularly imprinted polymer-based sensors for protein detection.

Science.gov (United States)

Whitcombe, Michael J; Chianella, Iva; Larcombe, Lee; Piletsky, Sergey A; Noble, James; Porter, Robert; Horgan, Adrian

2011-03-01

The detection of specific proteins as biomarkers of disease, health status, environmental monitoring, food quality, control of fermenters and civil defence purposes means that biosensors for these targets will become increasingly more important. Among the technologies used for building specific recognition properties, molecularly imprinted polymers (MIPs) are attracting much attention. In this critical review we describe many methods used for imprinting recognition for protein targets in polymers and their incorporation with a number of transducer platforms with the aim of identifying the most promising approaches for the preparation of MIP-based protein sensors (277 references).

50 Hz electric field effects on protein carbonyl (PCO), heme oxygenase-1 (HO-1) and hydroxyproline levels

International Nuclear Information System (INIS)

Ozgur, Elcin; Goknur, Guler; Seyhan, Nesrin

2008-01-01

Full text: Non-ionizing electromagnetic field (EMF) radiation sources, such as power lines and other Extremely Low Frequency (ELF) sources have become one of the most ubiquitous components of the spectrum of the human environment, and the possibility that they may have hazardous effects on human health is a major a public concern. Although it is well documented that EMFs have biological effects, the degree to which these exposures constitute a human health hazard is not clear yet. Today relation between production of oxidative stress resulted by reactive oxygen species and electrical stimulus, also the protective effects of antioxidant treatments are mentioned in many researches. In this study, it was aimed to determine both oxidation of proteins and protein collagen levels under 50 Hz 12 kV/m vertical Electric (E) Field exposure and the N-Acetylcysteine (NAC) administration which is a well-known antioxidant. To this end, protein carbonyl levels (PCO) as bio-markers of oxidative stress and Heme oxygenase-1 (HO-1), an enzyme that catalyzes the degradation of heme analyzed to figure out the protein oxidation. Hydroxyproline level, a major component of the protein collagen was measured in order to express the level of collagen in lung tissue. Guinea pigs, weighted 250-300 g, were used in the study. A total forty male guinea pigs were randomly divided into four groups which are composed of 10 guinea pigs each for groups: 1) Group I (Sham); 2) Group II (NAC-administrated group); 3) Group III (E Field Exposure group); 4) Group IV (NAC administrated + E Field exposed group). One week exposure period for 8 hours per daily was conducted for each exposure groups (Group III, Group IV ). The electric field exposure period was from 9 a.m. to 5 p.m. After the last exposure day, the guinea pigs were anesthetized by the injection of ketamine and xylazine. The guinea pigs were killed by decapitation. Statistical analyses were carried out using SPSS software (SPSS 11.5 for windows

Proteomic detection of oxidized and reduced thiol proteins in cultured cells.

Science.gov (United States)

Cuddihy, Sarah L; Baty, James W; Brown, Kristin K; Winterbourn, Christine C; Hampton, Mark B

2009-01-01

The oxidation and reduction of cysteine residues is emerging as an important post-translational control of protein function. We describe a method for fluorescent labelling of either reduced or oxidized thiols in combination with two-dimensional sodium dodecyl sulphate polyacrylamide gel electrophoresis (2DE) to detect changes in the redox proteome of cultured cells. Reduced thiols are labelled with the fluorescent compound 5-iodoacetamidofluorescein. To monitor oxidized thiols, the reduced thiols are first blocked with N-ethyl-maleimide, then the oxidized thiols reduced with dithiothreitol and labelled with 5-iodoacetamidofluorescein. The method is illustrated by treating Jurkat T-lymphoma cells with hydrogen peroxide and monitoring increased labelling of oxidized thiol proteins. A decrease in labelling can also be detected, and this is attributed to the formation of higher oxidation states of cysteine that are not reduced by dithiothreitol.

Ester carbonyl vibration as a sensitive probe of protein local electric field.

Science.gov (United States)

Pazos, Ileana M; Ghosh, Ayanjeet; Tucker, Matthew J; Gai, Feng

2014-06-10

The ability to quantify the local electrostatic environment of proteins and protein/peptide assemblies is key to gaining a microscopic understanding of many biological interactions and processes. Herein, we show that the ester carbonyl stretching vibration of two non-natural amino acids, L-aspartic acid 4-methyl ester and L-glutamic acid 5-methyl ester, is a convenient and sensitive probe in this regard, since its frequency correlates linearly with the local electrostatic field for both hydrogen-bonding and non-hydrogen-bonding environments. We expect that the resultant frequency-electric-field map will find use in various applications. Furthermore, we show that, when situated in a non-hydrogen-bonding environment, this probe can also be used to measure the local dielectric constant (ε). For example, its application to amyloid fibrils formed by Aβ(16-22) revealed that the interior of such β-sheet assemblies has an ε value of approximately 5.6. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Simultaneous detection of mRNA and protein stem cell markers in live cells

Directory of Open Access Journals (Sweden)

Bao Gang

2009-04-01

Full Text Available Abstract Background Biological studies and medical application of stem cells often require the isolation of stem cells from a mixed cell population, including the detection of cancer stem cells in tumor tissue, and isolation of induced pluripotent stem cells after eliciting the expression of specific genes in adult cells. Here we report the detection of Oct-4 mRNA and SSEA-1 protein in live carcinoma stem cells using respectively molecular beacon and dye-labeled antibody, aiming to establish a new method for stem cells detection and isolation. Results Quantification of Oct-4 mRNA and protein in P19 mouse carcinoma stem cells using respectively RT-PCR and immunocytochemistry confirmed that their levels drastically decreased after differentiation. To visualize Oct-4 mRNA in live stem cells, molecular beacons were designed, synthesized and validated, and the detection specificity was confirmed using control studies. We found that the fluorescence signal from Oct-4-targeting molecular beacons provides a clear discrimination between undifferentiated and retinoic acid-induced differentiated cells. Using deconvolution fluorescence microscopy, Oct-4 mRNAs were found to reside on one side of the cytosol. We demonstrated that, using a combination of Oct-4 mRNA-targeting molecular beacon with SSEA-1 antibody in flow cytometric analysis, undifferentiated stem cells can be clearly distinguished from differentiated cells. We revealed that Oct-4 targeting molecular beacons do not seem to affect stem cell biology. Conclusion Molecular beacons have the potential to provide a powerful tool for highly specific detection and isolation of stem cells, including cancer stem cells and induced pluripotent stem (iPS cells without disturbing cell physiology. It is advantageous to perform simultaneous detection of intracellular (mRNA and cell-surface (protein stem cell markers in flow cytometric analysis, which may lead to high detection sensitivity and efficiency.

Low-cost label-free electrical detection of artificial DNA nanostructures using solution-processed oxide thin-film transistors.

Science.gov (United States)

Kim, Si Joon; Jung, Joohye; Lee, Keun Woo; Yoon, Doo Hyun; Jung, Tae Soo; Dugasani, Sreekantha Reddy; Park, Sung Ha; Kim, Hyun Jae

2013-11-13

A high-sensitivity, label-free method for detecting deoxyribonucleic acid (DNA) using solution-processed oxide thin-film transistors (TFTs) was developed. Double-crossover (DX) DNA nanostructures with different concentrations of divalent Cu ion (Cu(2+)) were immobilized on an In-Ga-Zn-O (IGZO) back-channel surface, which changed the electrical performance of the IGZO TFTs. The detection mechanism of the IGZO TFT-based DNA biosensor is attributed to electron trapping and electrostatic interactions caused by negatively charged phosphate groups on the DNA backbone. Furthermore, Cu(2+) in DX DNA nanostructures generates a current path when a gate bias is applied. The direct effect on the electrical response implies that solution-processed IGZO TFTs could be used to realize low-cost and high-sensitivity DNA biosensors.

Practical photon number detection with electric field-modulated silicon avalanche photodiodes.

Science.gov (United States)

Thomas, O; Yuan, Z L; Shields, A J

2012-01-24

Low-noise single-photon detection is a prerequisite for quantum information processing using photonic qubits. In particular, detectors that are able to accurately resolve the number of photons in an incident light pulse will find application in functions such as quantum teleportation and linear optics quantum computing. More generally, such a detector will allow the advantages of quantum light detection to be extended to stronger optical signals, permitting optical measurements limited only by fluctuations in the photon number of the source. Here we demonstrate a practical high-speed device, which allows the signals arising from multiple photon-induced avalanches to be precisely discriminated. We use a type of silicon avalanche photodiode in which the lateral electric field profile is strongly modulated in order to realize a spatially multiplexed detector. Clearly discerned multiphoton signals are obtained by applying sub-nanosecond voltage gates in order to restrict the detector current.

Upconversion Nanoparticles-Encoded Hydrogel Microbeads-Based Multiplexed Protein Detection

Science.gov (United States)

Shikha, Swati; Zheng, Xiang; Zhang, Yong

2018-06-01

Fluorescently encoded microbeads are in demand for multiplexed applications in different fields. Compared to organic dye-based commercially available Luminex's xMAP technology, upconversion nanoparticles (UCNPs) are better alternatives due to their large anti-Stokes shift, photostability, nil background, and single wavelength excitation. Here, we developed a new multiplexed detection system using UCNPs for encoding poly(ethylene glycol) diacrylate (PEGDA) microbeads as well as for labeling reporter antibody. However, to prepare UCNPs-encoded microbeads, currently used swelling-based encapsulation leads to non-uniformity, which is undesirable for fluorescence-based multiplexing. Hence, we utilized droplet microfluidics to obtain encoded microbeads of uniform size, shape, and UCNPs distribution inside. Additionally, PEGDA microbeads lack functionality for probe antibodies conjugation on their surface. Methods to functionalize the surface of PEGDA microbeads (acrylic acid incorporation, polydopamine coating) reported thus far quench the fluorescence of UCNPs. Here, PEGDA microbeads surface was coated with silica followed by carboxyl modification without compromising the fluorescence intensity of UCNPs. In this study, droplet microfluidics-assisted UCNPs-encoded microbeads of uniform shape, size, and fluorescence were prepared. Multiple color codes were generated by mixing UCNPs emitting red and green colors at different ratios prior to encapsulation. UCNPs emitting blue color were used to label the reporter antibody. Probe antibodies were covalently immobilized on red UCNPs-encoded microbeads for specific capture of human serum albumin (HSA) as a model protein. The system was also demonstrated for multiplexed detection of both human C-reactive protein (hCRP) and HSA protein by immobilizing anti-hCRP antibodies on green UCNPs.

Titania nanotube-modified screen printed carbon electrodes enhance the sensitivity in the electrochemical detection of proteins.

Science.gov (United States)

Mandal, Soumit S; Navratna, Vikas; Sharma, Pratyush; Gopal, B; Bhattacharyya, Aninda J

2014-08-01

The use of titania nanotubes (TiO2-NT) as the working electrode provides a substantial improvement in the electrochemical detection of proteins. A biosensor designed using this strategy provided a robust method to detect protein samples at very low concentrations (Cprotein ca 1ng/μl). Reproducible measurements on protein samples at this concentration (Ip,a of 80+1.2μA) could be achieved using a sample volume of ca 30μl. We demonstrate the feasibility of this strategy for the accurate detection of penicillin binding protein, PBP2a, a marker for methicillin resistant Staphylococcus aureus (MRSA). The selectivity and efficiency of this sensor were also validated using other diverse protein preparations such as a recombinant protein tyrosine phosphatase (PTP10D) and bovine serum albumin (BSA). This electrochemical method also presents a substantial improvement in the time taken (few minutes) when compared to conventional enzyme-linked immunosorbent assay (ELISA) protocols. It is envisaged that this sensor could substantially aid in the rapid diagnosis of bacterial infections in resource strapped environments. Copyright © 2014 Elsevier B.V. All rights reserved.

Electrodeposition to construct free-standing chitosan/layered double hydroxides hydro-membrane for electrically triggered protein release.

Science.gov (United States)

Zhao, Pengkun; Zhao, Yanan; Xiao, Ling; Deng, Hongbing; Du, Yumin; Chen, Yun; Shi, Xiaowen

2017-10-01

In this study, we report the electrodeposition of a chitosan/layered double hydroxides (LDHs) hydro-membrane for protein release triggered by an electrical signal. The electrodeposition was performed in a chitosan and insulin loaded LDHs suspension in the absence of salt. A free-standing chitosan/LDHs hydro-membrane was generated on the electrode with improved mechanical properties, which is dramatically different from the weak hydrogel deposited in the presence of salt. The amount of LDHs in the hydro-membrane affects the optical transmittance and multilayered structure of the hybrid membrane. Compared to the weak chitosan/LDHs hydrogel, the hydro-membrane has a higher insulin loading capacity and the release of insulin is relatively slow. By biasing electrical potentials to the hydro-membrane, the release behavior of insulin can be adjusted accordingly. In addition, the chitosan/LDHs hydro-membrane showed no toxicity to cells. Our results provide a facile method to construct a chitosan/LDHs hybrid multilayered hydro-membrane and suggest the great potential of the hydro-membrane in controlled protein release. Copyright © 2017 Elsevier B.V. All rights reserved.

Detection and observation of deep vein thromboses by physical, radiological and electrical methods

International Nuclear Information System (INIS)

Bonafous, J.; Constantinesco, A.; Meyer, P.; Bidet, R.; Healy, J.C.; Chambron, J.; Lobera, A.; Chopin, J.

1975-01-01

The method of thrombosis detection by iodine 131-labelled exogenous fibrinogen was evaluated by comparison with three physical methods: an isotopic method using selenomethionine-labelled endogenous fibrinogen, phlebography, phletysmography by electrical impedance. The results confirm the superiority of the isotopic method with regard to efficiency and comfort for the patient. Under certain conditions the frequency of positive tests was the same with selenomethionine as with exogenous fibrinogen [fr

Highly multiplexed simultaneous detection of RNAs and proteins in single cells.

Science.gov (United States)

Frei, Andreas P; Bava, Felice-Alessio; Zunder, Eli R; Hsieh, Elena W Y; Chen, Shih-Yu; Nolan, Garry P; Gherardini, Pier Federico

2016-03-01

To enable the detection of expression signatures specific to individual cells, we developed PLAYR (proximity ligation assay for RNA), a method for highly multiplexed transcript quantification by flow and mass cytometry that is compatible with standard antibody staining. When used with mass cytometry, PLAYR allowed for the simultaneous quantification of more than 40 different mRNAs and proteins. In primary cells, we quantified multiple transcripts, with the identity and functional state of each analyzed cell defined on the basis of the expression of a separate set of transcripts or proteins. By expanding high-throughput deep phenotyping of cells beyond protein epitopes to include RNA expression, PLAYR opens a new avenue for the characterization of cellular metabolism.

Protein Profile study of clinical samples using Laser Induced Fluorescence as the detection method

DEFF Research Database (Denmark)

Karemore, Gopal Raghunath; Raja, Sujatha N.; Rai, Lavanya

2009-01-01

  Protein profiles of tissue homogenates were recorded using HPLC separation and LIF detection method. The samples were collected from volunteers with clinically normal or cervical cancer conditions. It is shown that the protein profile can be classified as belonging to malignant or normal state ...

Detection of Antibodies in Blood Plasma Using Bioluminescent Sensor Proteins and a Smartphone.

Science.gov (United States)

Arts, Remco; den Hartog, Ilona; Zijlema, Stefan E; Thijssen, Vito; van der Beelen, Stan H E; Merkx, Maarten

2016-04-19

Antibody detection is of fundamental importance in many diagnostic and bioanalytical assays, yet current detection techniques tend to be laborious and/or expensive. We present a new sensor platform (LUMABS) based on bioluminescence resonance energy transfer (BRET) that allows detection of antibodies directly in solution using a smartphone as the sole piece of equipment. LUMABS are single-protein sensors that consist of the blue-light emitting luciferase NanoLuc connected via a semiflexible linker to the green fluorescent acceptor protein mNeonGreen, which are kept close together using helper domains. Binding of an antibody to epitope sequences flanking the linker disrupts the interaction between the helper domains, resulting in a large decrease in BRET efficiency. The resulting change in color of the emitted light from green-blue to blue can be detected directly in blood plasma, even at picomolar concentrations of antibody. Moreover, the modular architecture of LUMABS allows changing of target specificity by simple exchange of epitope sequences, as demonstrated here for antibodies against HIV1-p17, hemagglutinin (HA), and dengue virus type I. The combination of sensitive ratiometric bioluminescent detection and the intrinsic modularity of the LUMABS design provides an attractive generic platform for point-of-care antibody detection that avoids the complex liquid handling steps associated with conventional immunoassays.

Electrical resistivity for detecting subsurface non-aqueous phase liquids: A progress report

International Nuclear Information System (INIS)

Lee, K.H.; Shan, C.; Javandel, I.

1995-06-01

Soils and groundwater have been contaminated by hazardous substances at many places in the United States and many other countries. The contaminants are commonly either petroleum products or industrial solvents with very low solubility in water. These contaminants are usually called non-aqueous phase liquids (NAPLs). The cost of cleaning up the affected sites in the United States is estimated to be of the order of 100 billion dollars. In spite of the expenditure of several billion dollars during the last 15 years, to date, very few, if any major contaminated site has been restored. The presence of NAPL pools in the subsurface is believed to be the main cause for the failure of previous cleanup activities. Due to their relatively low water solubility, and depending on their volume, it takes tens or even hundreds of years to deplete the NAPL sources if they are not removed from the subsurface. The intrinsic electrical resistivity of most NAPLs is typically in the range of 10 7 to 10 12 Ω-m, which is several orders of magnitude higher than that of groundwater containing dissolved solids (usually in the range of a few Ω-m to a few thousand Ω-m). Although a dry soil is very resistive, the electrical resistivity of a wet soil is on the order of 100 Ω-m and is dependent on the extent of water saturation. For a given soil, the electrical resistivity increases with decrease of water saturation. Therefore, if part of the pore water is replaced by a NAPL, the electrical resistivity will increase. At many NAPL sites, both the vadose and phreatic zones can be partially occupied by NAPL pools. It is the great contrast in electrical resistivity between the NAPLs and groundwater that may render the method to be effective in detecting subsurface NAPLs at contaminated sites. The following experiments were conducted to investigate the change of the electrical resistivity of porous media when diesel fuel (NAPL) replaces part of the water

Using an enzymatic galactose assay to detect lactose glycation extents of two proteins caseinate and soybean protein isolate via the Maillard reaction.

Science.gov (United States)

Wang, Xiao-Peng; Zhao, Xin-Huai

2017-06-01

Glycation of food proteins via the Maillard reaction has been widely studied in the recent years; however, the amount of saccharide connected to proteins is usually not determined. An enzymatic galactose assay was proposed firstly in this study to detect lactose glycation extents of caseinate and soybean protein isolate (SPI) during the Maillard reaction at two temperatures and different times. The separated glycated proteins were hydrolysed to release galactose necessary for the enzymatic assay and glycation calculation. Caseinate and SPI both obtained the highest lactose glycation extents at 100 °C or 121 °C by a reaction time of 180 or 20 min. Short- and long-time reaction resulted in lower glycation extents. During the reaction, three chemical indices (absorbences at 294/490 nm and fluorescence intensities) of reaction mixtures increased continually, but another index reactable NH 2 of glycated proteins showed the opposite trend. In general, changing profiles of the four indices were inconsistent with those profiles of lactose glycation extents of glycated proteins, implying practical limitation of the four indices in studies. This proposed enzymatic assay could directly detect lactose glycation of the two proteins, and thus was more useful than the four chemical indices to monitor glycation of the two proteins. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

Fluorescence detection of a protein-bound 2Fe2S cluster.

Science.gov (United States)

Hoff, Kevin G; Goodlitt, Rochelle; Li, Rui; Smolke, Christina D; Silberg, Jonathan J

2009-03-02

A fluorescent biosensor is described for 2Fe2S clusters that is composed of green fluorescent protein (GFP) fused to glutaredoxin 2 (Grx2), as illustrated here. 2Fe2S detection is based on the reduction of GFP fluorescence upon the 2Fe2S-induced dimerization of GFP-Grx2. This assay is sufficiently sensitive to detect submicromolar changes in 2Fe2S levels, thus making it suitable for high-throughput measurements of metallocluster degradation and synthesis reactions.

Sinkhole detection using electrical resistivity tomography in Saudi Arabia

International Nuclear Information System (INIS)

Youssef, Ahmed M; Zabramawi, Yasser A; El-Kaliouby, Hesham

2012-01-01

Karst phenomena exist in different areas in the Kingdom of Saudi Arabia, causing serious environmental problems that affect urban development and infrastructure (buildings, roads and highways). One of the most important problems are sinkholes, which most of the time consist of unfilled voids. These sinkholes are formed as a result of the chemical leaching of carbonate and evaporite formations by percolating water. Field investigations show that there are many surface expressions of sinkholes in the area; some appear on the ground surface and others are hidden in the subsurface. Geophysical data were collected at the study area using two-dimensional electrical resistivity tomography (ERT) with different electrode spacings to delineate buried sinkholes and associated subsurface cavities. Our findings indicated that the dipole–dipole method using an electrode spacing of 1 m was successful in detecting a known subsurface sinkhole. According to the ERT method the detected sinkhole depth ranges from 2 to 4 m, its height ranges from 2 to 4 m, and its width ranges from 5 to 7 m. Field observation has verified the geophysical data, especially along the profile A-A. Finally, closely spaced ERT profiles were successful in determining the three-dimensional volume of the subsurface sinkhole. (paper)

Nanoscale observation of local bound charges of patterned protein arrays by scanning force microscopy

International Nuclear Information System (INIS)

Oh, Y J; Jo, W; Kim, S; Park, S; Kim, Y S

2008-01-01

A protein patterned surface using micro-contact printing methods has been investigated by scanning force microscopy. Electrostatic force microscopy (EFM) was utilized for imaging the topography and detecting the electrical properties such as the local bound charge distribution of the patterned proteins. It was found that the patterned IgG proteins are arranged down to 1 μm, and the 90 deg. rotation of patterned anti-IgG proteins was successfully undertaken. Through the estimation of the effective areas, it was possible to determine the local bound charges of patterned proteins which have opposite electrostatic force behaviors. Moreover, we studied the binding probability between IgG and anti-IgG in a 1 μm 2 MIMIC system by topographic and electrostatic signals for applicable label-free detections. We showed that the patterned proteins can be used for immunoassay of proteins on the functional substrate, and that they can also be used for bioelectronics device application, indicating distinct advantages with regard to accuracy and a label-free detection

Identification of electric-field-dependent steps in the Na(+),K(+)-pump cycle.

Science.gov (United States)

Mares, Laura J; Garcia, Alvaro; Rasmussen, Helge H; Cornelius, Flemming; Mahmmoud, Yasser A; Berlin, Joshua R; Lev, Bogdan; Allen, Toby W; Clarke, Ronald J

2014-09-16

The charge-transporting activity of the Na(+),K(+)-ATPase depends on its surrounding electric field. To isolate which steps of the enzyme's reaction cycle involve charge movement, we have investigated the response of the voltage-sensitive fluorescent probe RH421 to interaction of the protein with BTEA (benzyltriethylammonium), which binds from the extracellular medium to the Na(+),K(+)-ATPase's transport sites in competition with Na(+) and K(+), but is not occluded within the protein. We find that only the occludable ions Na(+), K(+), Rb(+), and Cs(+) cause a drop in RH421 fluorescence. We conclude that RH421 detects intramembrane electric field strength changes arising from charge transport associated with conformational changes occluding the transported ions within the protein, not the electric fields of the bound ions themselves. This appears at first to conflict with electrophysiological studies suggesting extracellular Na(+) or K(+) binding in a high field access channel is a major electrogenic reaction of the Na(+),K(+)-ATPase. All results can be explained consistently if ion occlusion involves local deformations in the lipid membrane surrounding the protein occurring simultaneously with conformational changes necessary for ion occlusion. The most likely origin of the RH421 fluorescence response is a change in membrane dipole potential caused by membrane deformation. Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Aging Detection of Electrical Point Machines Based on Support Vector Data Description

Directory of Open Access Journals (Sweden)

Jaewon Sa

2017-11-01

Full Text Available Electrical point machines (EPM must be replaced at an appropriate time to prevent the occurrence of operational safety or stability problems in trains resulting from aging or budget constraints. However, it is difficult to replace EPMs effectively because the aging conditions of EPMs depend on the operating environments, and thus, a guideline is typically not be suitable for replacing EPMs at the most timely moment. In this study, we propose a method of classification for the detection of an aging effect to facilitate the timely replacement of EPMs. We employ support vector data description to segregate data of “aged” and “not-yet-aged” equipment by analyzing the subtle differences in normalized electrical signals resulting from aging. Based on the before and after-replacement data that was obtained from experimental studies that were conducted on EPMs, we confirmed that the proposed method was capable of classifying machines based on exhibited aging effects with adequate accuracy.

A network model to correlate conformational change and the impedance spectrum of single proteins

Energy Technology Data Exchange (ETDEWEB)

Alfinito, Eleonora; Pennetta, Cecilia; Reggiani, Lino [Dipartimento di Ingegneria dell' Innovazione, Universita del Salento, Via Arnesano, Lecce (Italy); Consorzio Nazionale Interuniversitario per le Scienze Fisiche della Materia (CNISM) (Italy)

2008-02-13

Integrated nanodevices based on proteins or biomolecules are attracting increasing interest in today's research. In fact, it has been shown that proteins such as azurin and bacteriorhodopsin manifest some electrical properties that are promising for the development of active components of molecular electronic devices. Here we focus on two relevant kinds of protein: bovine rhodopsin, prototype of G-protein-coupled-receptor (GPCR) proteins, and the enzyme acetylcholinesterase (AChE), whose inhibition is one of the most qualified treatments of Alzheimer's disease. Both these proteins exert their function starting with a conformational change of their native structure. Our guess is that such a change should be accompanied with a detectable variation of their electrical properties. To investigate this conjecture, we present an impedance network model of proteins, able to estimate the different impedance spectra associated with the different configurations. The distinct types of conformational change of rhodopsin and AChE agree with their dissimilar electrical responses. In particular, for rhodopsin the model predicts variations of the impedance spectra up to about 30%, while for AChE the same variations are limited to about 10%, which supports the existence of a dynamical equilibrium between its native and complexed states.

Detection of electrically neutral and nonpolar molecules in ionic solutions using silicon nanowires

Science.gov (United States)

Wu, Ying-Pin; Chu, Chia-Jung; Tsai, Li-Chu; Su, Ya-Wen; Chen, Pei-Hua; Moodley, Mathew K.; Huang, Ding; Chen, Yit-Tsong; Yang, Ying-Jay; Chen, Chii-Dong

2017-04-01

We report on a technique that can extend the use of nanowire sensors to the detection of interactions involving nonpolar and neutral molecules in an ionic solution environment. This technique makes use of the fact that molecular interactions result in a change in the permittivity of the molecules involved. For the interactions taking place at the surface of nanowires, this permittivity change can be determined from the analysis of the measured complex impedance of the nanowire. To demonstrate this technique, histidine was detected using different charge polarities controlled by the pH value of the solution. This included the detection of electrically neutral histidine at a sensitivity of 1 pM. Furthermore, it is shown that nonpolar molecules, such as hexane, can also be detected. The technique is applicable to the use of nanowires with and without a surface-insulating oxide. We show that information about the changes in amplitude and the phase of the complex impedance reveals the fundamental characteristics of the molecular interactions, including the molecular field and the permittivity.

Specific, sensitive, high-resolution detection of protein molecules in eukaryotic cells using metal-tagging transmission electron microscopy

Science.gov (United States)

Risco, Cristina; Sanmartín-Conesa, Eva; Tzeng, Wen-Pin; Frey, Teryl K.; Seybold, Volker; de Groot, Raoul J.

2012-01-01

Summary More than any other methodology, transmission electron microscopy (TEM) has contributed to our understanding of the architecture and organization of cells. With current detection limits approaching atomic resolution, it will ultimately become possible to ultrastructurally image intracellular macromolecular assemblies in situ. Presently, however, methods to unambiguously identify proteins within the crowded environment of the cell’s interior are lagging behind. We describe a novel approach, metal-tagging TEM (METTEM) that allows detection of intracellular proteins in mammalian cells with high specificity, exceptional sensitivity and at molecular scale resolution. In live cells treated with gold salts, proteins bearing a small metal-binding tag will form 1-nm gold nanoclusters, readily detectable in electron micrographs. The applicability and strength of METTEM is demonstrated by a study of Rubella virus replicase and capsid proteins, which revealed virus-induced cell structures not seen before. PMID:22579245

Detection of irradiated food by the changes in protein molecular mass distribution

International Nuclear Information System (INIS)

Niciforovic, A.; Radojcic, M.; Milosavljevic, B.H.

1998-01-01

Complete text of publication follows. The present work deals with the radiation-induced damage of proteins, which is followed by the change in the molecular mass. The phenomenon was studied on protein rich samples, i.e., chicken meat and dehydrated egg white. The radiation dose applied was in the range of the ones used for food microbial control. Chicken drumstick and chicken white meat proteins were separated according to their molecular mass. The protein profile was compared to the meat samples irradiated in the frozen state with 5 kGy at 60 Co source. In the case of chicken white meat, irradiation produces both nonselective protein scission (e.g. the amount of proteins of molecular mass larger than 30 kDa decreases, while the amount of proteins of molecular mass smaller than 30 kDa increases), and selective protein scission (e.g. appearance of a protein fragment of molecular mass equal to 18 kDa). In the case of chicken drumstick proteins the irradiation induces both the protein scission and the aggregation. The changes are nonspecific as well as specific and the generation of Mm = 18 kDa protein fragment was observed again. Irradiation of aerated dehydrated egg white proteins produces only nonselective protein scission. The results are discussed in view of the routine application of SDS-PAGE method for the detection of irradiated foodstuff

Modification of ISFETs with a monolayer of latex beads for specific detection of proteins

NARCIS (Netherlands)

Besselink, G.A.J.; Schasfoort, Richardus B.M.; Bergveld, Piet

2003-01-01

The so-called ion-step method is a novel potentiometric approach that can detect protein adsorbed onto the gate area of modified ion-sensitive field-effect transistors (ISFETs). In this report, a generic technology is described for immobilization of peptides and proteins to the ISFET gate in order

Modifying a standard method allows simultaneous extraction of RNA and protein, enabling detection of enzymes in the rat retina with low expressions and protein levels.

Science.gov (United States)

Agardh, Elisabet; Gustavsson, Carin; Hagert, Per; Nilsson, Marie; Agardh, Carl-David

2006-02-01

The aim of the study was to evaluate messenger RNA and protein expression in limited amounts of tissue with low protein content. The Chomczynski method was used for simultaneous extraction of RNA, and protein was modified in the protein isolation step. Template mass and cycling time for the complementary DNA synthesis step of real-time reverse transcription-polymerase chain reaction (RT-PCR) for analysis of catalase, copper/zinc superoxide dismutase, manganese superoxide dismutase, the catalytic subunit of glutamylcysteine ligase, glutathione peroxidase 1, and the endogenous control cyclophilin B (CypB) were optimized before PCR. Polymerase chain reaction accuracy and efficacy were demonstrated by calculating the regression (R2) values of the separate amplification curves. Appropriate antibodies, blocking buffers, and running conditions were established for Western blot, and protein detection and multiplex assays with CypB were performed for each target. During the extraction procedure, the protein phase was dissolved in a modified washing buffer containing 0.1% sodium dodecyl sulfate, followed by ultrafiltration. Enzyme expression on real-time RT-PCR was accomplished with high reliability and reproducibility (R2, 0.990-0.999), and all enzymes except for glutathione peroxidase 1 were detectable in individual retinas on Western blot. Western blot multiplexing with CypB was possible for all targets. In conclusion, connecting gene expression directly to protein levels in the individual rat retina was possible by simultaneous extraction of RNA and protein. Real-time RT-PCR and Western blot allowed accurate detection of retinal protein expressions and levels.

Direct, Specific and Rapid Detection of Staphylococcal Proteins and Exotoxins Using a Multiplex Antibody Microarray.

Directory of Open Access Journals (Sweden)

Bettina Stieber

Full Text Available S. aureus is a pathogen in humans and animals that harbors a wide variety of virulence factors and resistance genes. This bacterium can cause a wide range of mild to life-threatening diseases. In the latter case, fast diagnostic procedures are important. In routine diagnostic laboratories, several genotypic and phenotypic methods are available to identify S. aureus strains and determine their resistances. However, there is a demand for multiplex routine diagnostic tests to directly detect staphylococcal toxins and proteins.In this study, an antibody microarray based assay was established and validated for the rapid detection of staphylococcal markers and exotoxins. The following targets were included: staphylococcal protein A, penicillin binding protein 2a, alpha- and beta-hemolysins, Panton Valentine leukocidin, toxic shock syndrome toxin, enterotoxins A and B as well as staphylokinase. All were detected simultaneously within a single experiment, starting from a clonal culture on standard media. The detection of bound proteins was performed using a new fluorescence reading device for microarrays.110 reference strains and clinical isolates were analyzed using this assay, with a DNA microarray for genotypic characterization performed in parallel. The results showed a general high concordance of genotypic and phenotypic data. However, genotypic analysis found the hla gene present in all S. aureus isolates but its expression under given conditions depended on the clonal complex affiliation of the actual isolate.The multiplex antibody assay described herein allowed a rapid and reliable detection of clinically relevant staphylococcal toxins as well as resistance- and species-specific markers.

Detection of cow's milk proteins and minor components in human milk using proteomics techniques.

Science.gov (United States)

Coscia, A; Orrù, S; Di Nicola, P; Giuliani, F; Varalda, A; Peila, C; Fabris, C; Conti, A; Bertino, E

2012-10-01

Cow's milk proteins (CMPs) are the best characterized food allergens. The aim of this study was to investigate cow's milk allergens in human colostrum of term and preterm newborns' mothers, and other minor protein components by proteomics techniques, more sensitive than other techniques used in the past. Sixty-two term and 11 preterm colostrum samples were collected, subjected to a treatment able to increase the concentration of the most diluted proteins and simultaneously to reduce the concentration of the proteins present at high concentration (Proteominer Treatment), and subsequently subjected to the steps of proteomic techniques. The most relevant finding in this study was the detection of the intact bovine alpha-S1-casein in human colostrum, then bovine alpha-1-casein could be considered the cow's milk allergen that is readily secreted in human milk and could be a cause of sensitization to cow's milk in exclusively breastfed predisposed infants. Another interesting result was the detection, at very low concentrations, of proteins previously not described in human milk (galectin-7, the different isoforms of the 14-3-3 protein and the serum amyloid P-component), probably involved in the regulation of the normal cell growth, in the pro-apoptotic function and in the regulation of tissue homeostasis. Further investigations are needed to understand if these families of proteins have specific biological activity in human milk.

An electrical bio-chip to transfer and detect electromagnetic stimulation on the cells based on vertically aligned carbon nanotubes

Energy Technology Data Exchange (ETDEWEB)

Rafizadeh-Tafti, Saeed [Nanoelectronic Center of Excellence, Thin Film and Nanoelectronic Lab, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395/515, Tehran (Iran, Islamic Republic of); Nano Bio Electronic Devices Lab, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395/515, Tehran (Iran, Islamic Republic of); Haqiqatkhah, Mohammad Hossein [Center of Excellence on Applied Electromagnetic Systems, School of Electrical & Computer Engineering, University of Tehran, P.O. Box 14395-515, North Kargar Avenue, Tehran (Iran, Islamic Republic of); Saviz, Mehrdad [Antenna Laboratory, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395-515, North Kargar Avenue, Tehran (Iran, Islamic Republic of); Janmaleki, Mohsen [Medical Nanotechnology and Tissue Engineering Research Center, Shahid Beheshti University of Medical Sciences, P.O. Box 1985717443, Tehran (Iran, Islamic Republic of); Faraji Dana, Reza [Center of Excellence on Applied Electromagnetic Systems, School of Electrical & Computer Engineering, University of Tehran, P.O. Box 14395-515, North Kargar Avenue, Tehran (Iran, Islamic Republic of); Zanganeh, Somayeh [Nanoelectronic Center of Excellence, Thin Film and Nanoelectronic Lab, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395/515, Tehran (Iran, Islamic Republic of); Nano Bio Electronic Devices Lab, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395/515, Tehran (Iran, Islamic Republic of); Abdolahad, Mohammad, E-mail: m.abdolahad@ut.ac.ir [Nanoelectronic Center of Excellence, Thin Film and Nanoelectronic Lab, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395/515, Tehran (Iran, Islamic Republic of); Nano Bio Electronic Devices Lab, School of Electrical and Computer Engineering, University of Tehran, P.O. Box 14395/515, Tehran (Iran, Islamic Republic of)

2017-01-01

A highly sensitive impedimetric bio-chip based on vertically aligned multiwall carbon nanotubes (VAMWCNTs), was applied in direct interaction with lung cancer cells. Our tool provided both inducing and monitoring the bioelectrical changes in the cells initiated by electromagnetic (EM) wave stimulation. EM wave of 940 MHz frequency with different intensities was used. Here, wave ablation might accumulate electrical charge on the tips of nanotubes penetrated into cell's membrane. The charge might induce ionic exchanges into the cell and cause alterations in electrical states of the membrane. Transmembrane electrostatic/dynamic states would be strongly affected due to such exchanges. Our novel modality was that, the cells' vitality changes caused by charge inductions were electrically detected with the same nanotubes in the architecture of electrodes for impedance measurement. The responses of the sensor were confirmed by electron and florescent microscopy images as well as biological assays. In summation, our method provided an effective biochip for enhancing and detecting external EM stimulation on the cells useful for future diagnostic and therapeutic applications, such as wave-guided drug-resistance breakage. - Highlights: • A CNT-chip is fabricated to stimulate cancer cells by electromagnetic wave. • Wave induced charges accumulation on the tip of CNTs penetrated into cells. • Transmembrane electrostatic states would be strongly affected due to such exchanges. • The cells' vitality changes could be happened and electrically detected with the same chip.

Detection of tumor cell-specific mRNA and protein in exosome-like microvesicles from blood and saliva.

Science.gov (United States)

Yang, Jieping; Wei, Fang; Schafer, Christopher; Wong, David T W

2014-01-01

The discovery of disease-specific biomarkers in oral fluids has revealed a new dimension in molecular diagnostics. Recent studies have reported the mechanistic involvement of tumor cells derived mediators, such as exosomes, in the development of saliva-based mRNA biomarkers. To further our understanding of the origins of disease-induced salivary biomarkers, we here evaluated the hypothesis that tumor-shed secretory lipidic vesicles called exosome-like microvesicles (ELMs) that serve as protective carriers of tissue-specific information, mRNAs, and proteins, throughout the vasculature and bodily fluids. RNA content was analyzed in cell free-saliva and ELM-enriched fractions of saliva. Our data confirmed that the majority of extracellular RNAs (exRNAs) in saliva were encapsulated within ELMs. Nude mice implanted with human lung cancer H460 cells expressing hCD63-GFP were used to follow the circulation of tumor cell specific protein and mRNA in the form of ELMs in vivo. We were able to identify human GAPDH mRNA in ELMs of blood and saliva of tumor bearing mice using nested RT-qPCR. ELMs positive for hCD63-GFP were detected in the saliva and blood of tumor bearing mice as well as using electric field-induced release and measurement (EFIRM). Altogether, our results demonstrate that ELMs carry tumor cell-specific mRNA and protein from blood to saliva in a xenografted mouse model of human lung cancer. These results therefore strengthen the link between distal tumor progression and the biomarker discovery of saliva through the ELMs.

Detection of tumor cell-specific mRNA and protein in exosome-like microvesicles from blood and saliva.

Directory of Open Access Journals (Sweden)

Jieping Yang

Full Text Available The discovery of disease-specific biomarkers in oral fluids has revealed a new dimension in molecular diagnostics. Recent studies have reported the mechanistic involvement of tumor cells derived mediators, such as exosomes, in the development of saliva-based mRNA biomarkers. To further our understanding of the origins of disease-induced salivary biomarkers, we here evaluated the hypothesis that tumor-shed secretory lipidic vesicles called exosome-like microvesicles (ELMs that serve as protective carriers of tissue-specific information, mRNAs, and proteins, throughout the vasculature and bodily fluids. RNA content was analyzed in cell free-saliva and ELM-enriched fractions of saliva. Our data confirmed that the majority of extracellular RNAs (exRNAs in saliva were encapsulated within ELMs. Nude mice implanted with human lung cancer H460 cells expressing hCD63-GFP were used to follow the circulation of tumor cell specific protein and mRNA in the form of ELMs in vivo. We were able to identify human GAPDH mRNA in ELMs of blood and saliva of tumor bearing mice using nested RT-qPCR. ELMs positive for hCD63-GFP were detected in the saliva and blood of tumor bearing mice as well as using electric field-induced release and measurement (EFIRM. Altogether, our results demonstrate that ELMs carry tumor cell-specific mRNA and protein from blood to saliva in a xenografted mouse model of human lung cancer. These results therefore strengthen the link between distal tumor progression and the biomarker discovery of saliva through the ELMs.

A cell-based fluorescent assay to detect the activity of AB toxins that inhibit protein synthesis

Science.gov (United States)

AB-type protein toxins, produced by numerous bacterial pathogens and some plants, elicit a cytotoxic effect involving the inhibition of protein synthesis. To develop an improved method to detect the inhibition of protein synthesis by AB-type toxins, the present study characterized a Vero cell line t...

Detecting Protein-Glycolipid Interactions Using Glycomicelles and CaR-ESI-MS.

Science.gov (United States)

Han, Ling; Kitova, Elena N; Klassen, John S

2016-11-01

This study reports on the use of the catch-and-release electrospray ionization mass spectrometry (CaR-ESI-MS) assay, combined with glycomicelles, as a method for detecting specific interactions between water-soluble proteins and glycolipids (GLs) in aqueous solution. The B subunit homopentamers of cholera toxin (CTB 5 ) and Shiga toxin type 1 B (Stx1B 5 ) and the gangliosides GM1, GM2, GM3, GD1a, GD1b, GT1b, and GD2 served as model systems for this study. The CTB 5 exhibits broad specificity for gangliosides and binds to GM1, GM2, GM3, GD1a, GD1b, and GT1b; Stx1B 5 does not recognize gangliosides. The CaR-ESI-MS assay was used to analyze solutions of CTB 5 or Stx1B 5 and individual gangliosides (GM1, GM2, GM3, GD1a, GD1b, GT1b, and GD2) or mixtures thereof. The high affinity interaction of CTB 5 with GM1 was successfully detected. However, the apparent affinity, as determined from the mass spectra, is significantly lower than that of the corresponding pentasaccharide or when GM1 is presented in model membranes such as nanodiscs. Interactions between CTB 5 and the low affinity gangliosides GD1a, GD1b, and GT1b, as well as GD2, which served as a negative control, were detected; no binding of CTB 5 to GM2 or GM3 was observed. The CaR-ESI-MS results obtained for Stx1B 5 reveal that nonspecific protein-ganglioside binding can occur during the ESI process, although the extent of binding varies between gangliosides. Consequently, interactions detected for CTB 5 with GD1a, GD1b, and GT1b are likely nonspecific in origin. Taken together, these results reveal that the CaR-ESI-MS/glycomicelle approach for detecting protein-GL interactions is prone to false positives and false negatives and must be used with caution. Graphical Abstract GRAPHICAL ABSTRACT TEXT HERE] -->.

Graphene-interfaced electrical biosensor for label-free and sensitive detection of foodborne pathogenic E. coli O157:H7.

Science.gov (United States)

Pandey, Ashish; Gurbuz, Yasar; Ozguz, Volkan; Niazi, Javed H; Qureshi, Anjum

2017-05-15

E. coli O157:H7 is an enterohemorrhagic bacteria responsible for serious foodborne outbreaks that causes diarrhoea, fever and vomiting in humans. Recent foodborne E. coli outbreaks has left a serious concern to public health. Therefore, there is an increasing demand for a simple, rapid and sensitive method for pathogen detection in contaminated foods. In this study, we developed a label-free electrical biosensor interfaced with graphene for sensitive detection of pathogenic bacteria. This biosensor was fabricated by interfacing graphene with interdigitated microelectrodes of capacitors that were biofunctionalized with E. coli O157:H7 specific antibodies for sensitive pathogenic bacteria detection. Here, graphene nanostructures on the sensor surface provided superior chemical properties such as high carrier mobility and biocompatibility with antibodies and bacteria. The sensors transduced the signal based on changes in dielectric properties (capacitance) through (i) polarization of captured cell-surface charges, (ii) cells' internal bioactivity, (iii) cell-wall's electronegativity or dipole moment and their relaxation and (iv) charge carrier mobility of graphene that modulated the electrical properties once the pathogenic E. coli O157:H7 captured on the sensor surface. Sensitive capacitance changes thus observed with graphene based capacitors were specific to E. coli O157:H7 strain with a sensitivity as low as 10-100 cells/ml. The proposed graphene based electrical biosensor provided advantages of speed, sensitivity, specificity and in-situ bacterial detection with no chemical mediators, represents a versatile approach for detection of a wide variety of other pathogens. Copyright © 2016 Elsevier B.V. All rights reserved.

Endogenous field feedback promotes the detectability for exogenous electric signal in the hybrid coupled population

Energy Technology Data Exchange (ETDEWEB)

Wei, Xile; Zhang, Danhong; Wang, Jiang; Yu, Haitao, E-mail: htyu@tju.edu.cn [Tianjin Key Laboratory of Process Measurement and Control, School of Electrical Engineering and Automation, Tianjin University, Tianjin 300072 (China); Lu, Meili [School of Informational Technology and Engineering, Tianjin University of Technology and Education, Tianjin 300222 (China); Che, Yanqiu [School of Automation and Electrical Engineering, Tianjin University of Technology and Education, Tianjin 300222 (China)

2015-01-15

This paper presents the endogenous electric field in chemical or electrical synaptic coupled networks, aiming to study the role of endogenous field feedback in the signal propagation in neural systems. It shows that the feedback of endogenous fields to network activities can reduce the required energy of the noise and enhance the transmission of input signals in hybrid coupled populations. As a common and important nonsynaptic interactive method among neurons, particularly, the endogenous filed feedback can not only promote the detectability of exogenous weak signal in hybrid coupled neural population but also enhance the robustness of the detectability against noise. Furthermore, with the increasing of field coupling strengths, the endogenous field feedback is conductive to the stochastic resonance by facilitating the transition of cluster activities from the no spiking to spiking regions. Distinct from synaptic coupling, the endogenous field feedback can play a role as internal driving force to boost the population activities, which is similar to the noise. Thus, it can help to transmit exogenous weak signals within the network in the absence of noise drive via the stochastic-like resonance.

Endogenous field feedback promotes the detectability for exogenous electric signal in the hybrid coupled population.

Science.gov (United States)

Wei, Xile; Zhang, Danhong; Lu, Meili; Wang, Jiang; Yu, Haitao; Che, Yanqiu

2015-01-01

This paper presents the endogenous electric field in chemical or electrical synaptic coupled networks, aiming to study the role of endogenous field feedback in the signal propagation in neural systems. It shows that the feedback of endogenous fields to network activities can reduce the required energy of the noise and enhance the transmission of input signals in hybrid coupled populations. As a common and important nonsynaptic interactive method among neurons, particularly, the endogenous filed feedback can not only promote the detectability of exogenous weak signal in hybrid coupled neural population but also enhance the robustness of the detectability against noise. Furthermore, with the increasing of field coupling strengths, the endogenous field feedback is conductive to the stochastic resonance by facilitating the transition of cluster activities from the no spiking to spiking regions. Distinct from synaptic coupling, the endogenous field feedback can play a role as internal driving force to boost the population activities, which is similar to the noise. Thus, it can help to transmit exogenous weak signals within the network in the absence of noise drive via the stochastic-like resonance.

Endogenous field feedback promotes the detectability for exogenous electric signal in the hybrid coupled population

International Nuclear Information System (INIS)

Wei, Xile; Zhang, Danhong; Wang, Jiang; Yu, Haitao; Lu, Meili; Che, Yanqiu

2015-01-01

This paper presents the endogenous electric field in chemical or electrical synaptic coupled networks, aiming to study the role of endogenous field feedback in the signal propagation in neural systems. It shows that the feedback of endogenous fields to network activities can reduce the required energy of the noise and enhance the transmission of input signals in hybrid coupled populations. As a common and important nonsynaptic interactive method among neurons, particularly, the endogenous filed feedback can not only promote the detectability of exogenous weak signal in hybrid coupled neural population but also enhance the robustness of the detectability against noise. Furthermore, with the increasing of field coupling strengths, the endogenous field feedback is conductive to the stochastic resonance by facilitating the transition of cluster activities from the no spiking to spiking regions. Distinct from synaptic coupling, the endogenous field feedback can play a role as internal driving force to boost the population activities, which is similar to the noise. Thus, it can help to transmit exogenous weak signals within the network in the absence of noise drive via the stochastic-like resonance

Enhancing the lateral-flow immunoassay for detection of proteins using an aqueous two-phase micellar system.

Science.gov (United States)

Mashayekhi, Foad; Le, Alexander M; Nafisi, Parsa M; Wu, Benjamin M; Kamei, Daniel T

2012-10-01

The lateral-flow (immuno)assay (LFA) has been widely investigated for the detection of molecular, macromolecular, and particle targets at the point-of-need due to its ease of use, rapid processing, and minimal power and laboratory equipment requirements. However, for some analytes, such as certain proteins, the detection limit of LFA is inferior to lab-based assays, such as the enzyme-linked immunosorbent assay, and needs to be improved. One solution for improving the detection limit of LFA is to concentrate the target protein in a solution prior to the detection step. In this study, a novel approach was used in the context of an aqueous two-phase micellar system comprised of the nonionic surfactant Triton X-114 to concentrate a model protein, namely transferrin, prior to LFA. Proteins have been shown to partition, or distribute, fairly evenly between the two phases of an aqueous two-phase system, which in turn results in their limited concentration in one of the two phases. Therefore, larger colloidal gold particles decorated with antibodies for transferrin were used in the concentration step to bind to transferrin and aid its partitioning into the top, micelle-poor phase. By manipulating the volume ratio of the two coexisting micellar phases and combining the concentration step with LFA, the transferrin detection limit of LFA was improved by tenfold from 0.5 to 0.05 μg/mL in a predictive manner. In addition to enhancing the sensitivity of LFA, this universal concentration method could also be used to improve other detection assays.

Evaluating the efficacy of a structure-derived amino acid substitution matrix in detecting protein homologs by BLAST and PSI-BLAST.

Science.gov (United States)

Goonesekere, Nalin Cw

2009-01-01

The large numbers of protein sequences generated by whole genome sequencing projects require rapid and accurate methods of annotation. The detection of homology through computational sequence analysis is a powerful tool in determining the complex evolutionary and functional relationships that exist between proteins. Homology search algorithms employ amino acid substitution matrices to detect similarity between proteins sequences. The substitution matrices in common use today are constructed using sequences aligned without reference to protein structure. Here we present amino acid substitution matrices constructed from the alignment of a large number of protein domain structures from the structural classification of proteins (SCOP) database. We show that when incorporated into the homology search algorithms BLAST and PSI-blast, the structure-based substitution matrices enhance the efficacy of detecting remote homologs.

Fault detection and classification in electrical power transmission system using artificial neural network.

Science.gov (United States)

Jamil, Majid; Sharma, Sanjeev Kumar; Singh, Rajveer

2015-01-01

This paper focuses on the detection and classification of the faults on electrical power transmission line using artificial neural networks. The three phase currents and voltages of one end are taken as inputs in the proposed scheme. The feed forward neural network along with back propagation algorithm has been employed for detection and classification of the fault for analysis of each of the three phases involved in the process. A detailed analysis with varying number of hidden layers has been performed to validate the choice of the neural network. The simulation results concluded that the present method based on the neural network is efficient in detecting and classifying the faults on transmission lines with satisfactory performances. The different faults are simulated with different parameters to check the versatility of the method. The proposed method can be extended to the Distribution network of the Power System. The various simulations and analysis of signals is done in the MATLAB(®) environment.

Detection and properties of A-factor-binding protein from Streptomyces griseus

International Nuclear Information System (INIS)

Miyake, K.; Horinouchi, S.; Yoshida, M.; Chiba, N.; Mori, K.; Nogawa, N.; Morikawa, N.; Beppu, T.

1989-01-01

The optically active form of tritium-labeled A-factor (2-isocapryloyl-3R-hydroxymethyl-gamma-butyrolactone), a pleiotropic autoregulator responsible for streptomycin production, streptomycin resistance, and sporulation in Streptomyces griseus, was chemically synthesized. By using the radioactive A-factor, a binding protein for A-factor was detected in the cytoplasmic fraction of this organism. The binding protein had an apparent molecular weight of approximately 26,000, as determined by gel filtration. Scatchard analysis suggested that A-factor bound the protein in the molar ratio of 1:1 with a binding constant, Kd, of 0.7 nM. The number of the binding protein was roughly estimated to be 37 per genome. The inducing material virginiae butanolide C (VB-C), which has a structure very similar to that of A-factor and is essential for virginiamycin production in Streptomyces virginiae, did not inhibit binding. In addition, no protein capable of specifically binding 3 H-labeled VB-C was found in S. griseus. Together with the observation that VB-C had almost no biological activity on the restoration of streptomycin production or sporulation in an A-factor-deficient mutant of S. griseus, these results indicated that the binding protein had a strict ligand specificity. Examination for an A-factor-binding protein in Streptomyces coelicolor A3(2) and Streptomyces lividans showed the absence of any specifically binding protein

Dynamics of a magnetic monopole in matter, Maxwell equations in dyonic matter and detection of electric dipole moments

International Nuclear Information System (INIS)

Artru, X.; Fayolle, D.

2001-01-01

For a monopole, the analogue of the Lorentz equation in matter is shown to be f = g (H-v centre dot D). Dual-symmetric Maxwell equations, for matter containing hidden magnetic charge in addition to electric ones, are given. They apply as well to ordinary matter if the particles possess T-violating electric dipole moments. Two schemes of experiments for the detection of such moments in macroscopic pieces of matter are proposed

Perturbation of hydration layer in solvated proteins by external electric and electromagnetic fields: Insights from non-equilibrium molecular dynamics

Science.gov (United States)

Nandi, Prithwish K.; Futera, Zdenek; English, Niall J.

2016-11-01

Given the fundamental role of water in governing the biochemistry of enzymes, and in regulating their wider biological activity (e.g., by local water concentration surrounding biomolecules), the influence of extraneous electric and electromagnetic (e/m) fields thereon is of central relevance to biophysics and, more widely, biology. With the increase in levels of local and atmospheric microwave-frequency radiation present in modern life, as well as other electric-field exposure, the impact upon hydration-water layers surrounding proteins, and biomolecules generally, becomes a particularly pertinent issue. Here, we present a (non-equilibrium) molecular-dynamics-simulation study on a model protein (hen egg-white lysozyme) hydrated in water, in which we determine, inter alia, translational self-diffusivities for both hen egg-white lysozyme and its hydration layer together with relaxation dynamics of the hydrogen-bond network between the protein and its hydration-layer water molecules on a residue-per-residue basis. Crucially, we perform this analysis both above and below the dynamical-transition temperature (at ˜220 K), at 300 and 200 K, respectively, and we compare the effects of external static-electric and e/m fields with linear-response-régime (r.m.s.) intensities of 0.02 V/Å. It was found that the translational self-diffusivity of hen egg-white lysozyme and its hydration-water layer are increased substantially in static fields, primarily due to the induced electrophoretic motion, whilst the water-protein hydrogen-bond-network-rearrangement kinetics can also undergo rather striking accelerations, primarily due to the enhancement of a larger-amplitude local translational and rotational motion by charged and dipolar residues, which serves to promote hydrogen-bond breakage and re-formation kinetics. These external-field effects are particularly evident at 200 K, where they serve to induce the protein- and solvation-layer-response effects redolent of dynamical

Using a silver-enhanced microarray sandwich structure to improve SERS sensitivity for protein detection.

Science.gov (United States)

Gu, Xuefang; Yan, Yuerong; Jiang, Guoqing; Adkins, Jason; Shi, Jian; Jiang, Guomin; Tian, Shu

2014-03-01

A simple and sensitive method, based on surface-enhanced Raman scattering (SERS), for immunoassay and label-free protein detection is reported. A series of bowl-shaped silver cavity arrays were fabricated by electrodeposition using a self-assembled polystyrene spheres template. The reflection spectra of these cavity arrays were recorded as a function of film thickness, and then correlated with SERS enhancement using sodium thiophenolate as the probe molecule. The results reveal that SERS enhancement can be maximized when the frequency of both the incident laser and the Raman scattering approach the frequency of the localized surface plasmon resonance. The optimized array was then used as the bottom layer of a silver nanoparticle-protein-bowl-shaped silver cavity array sandwich. The second layer of silver was introduced by the interactions between the proteins in the middle layer of the sandwich architecture and silver nanoparticles. Human IgG bound to the surface of this microcavity array can retain its recognition function. With the Raman reporter molecules labeled on the antibody, a detection limit down to 0.1 ng mL(-1) for human IgG is easily achieved. Furthermore, the SERS spectra of label-free proteins (catalase, cytochrome C, avidin and lysozyme) from the assembled sandwich have excellent reproducibility and high quality. The results reveal that the proposed approach has potential for use in qualitative and quantitative detection of biomolecules.

Detection of hidden hazelnut protein in food by IgY-based indirect competitive enzyme-immunoassay

NARCIS (Netherlands)

Baumgartner, S.; Bremer, M.G.E.G.; Kemmers - Voncken, A.E.M.; Smits, N.G.E.; Haasnoot, W.; Banks, J.; Reece, P.; Danks, C.; Tomkies, V.; Immer, U.; Schmitt, K.; Krska, R.

2004-01-01

The development of an indirect competitive enzyme-immunoassay for the detection of hidden hazelnut protein in complex food matrices is described. A sensitive and selective polyclonal antibody was raised by immunisation of laying hens with protein extracts from roasted hazelnuts. In contrast to

Evaluation of reconstruction parameters of electrical impedance tomography on aorta detection during saline bolus injection

Directory of Open Access Journals (Sweden)

Thürk Florian

2016-09-01

Full Text Available An accurate detection of anatomical structures in electrical impedance tomography (EIT is still at an early stage. Aorta detection in EIT is of special interest, since it would favor non-invasive assessment of hemodynamic processes in the body. Here, diverse EIT reconstruction parameters of the GREIT algorithm were systematically evaluated to detect the aorta after saline bolus injection in apnea. True aorta position and size were taken from computed tomography (CT. A comparison with CT showed that the smallest error for aorta displacement was attained for noise figure nf = 0.7, weighting radius rw = 0.15, and target size ts = 0.01. The spatial extension of the aorta was most precise for nf = 0.7, rw = 0.25, and ts = 0.07. Detection accuracy (F1-score was highest with nf = 0.6, rw = 0.15, and ts = 0.04. This work provides algorithm-related evidence for potentially accurate aorta detection in EIT after injection of a saline bolus.

Application of Electrical Resistivity Tomography for Detecting Root Biomass in Coffee Trees

Directory of Open Access Journals (Sweden)

Carlos Mauricio Paglis

2013-01-01

Full Text Available Roots play an important role in plants and are responsible for several functions; among them are anchorage and nutrient and water absorption. Several methodologies are being tested and used to study plant root systems in order to avoid destructive root sampling. Electrical resistivity tomography is among these methodologies. The aim of this preliminary study was to use electrical resistivity for detecting root biomass in coffee trees. Measurements were performed in a soil transect with an ABM AL 48-b resistivimeter with a pole-dipole configuration. The tomograms indicated variability in soil resistivity values ranging from 120 to 1400 Ω·m−1. At the first 0.30 cm soil layer, these values were between 267 and 952 Ω·m−1. Oriented by this result, root samples were taken at 0.10, 0.20, and 0.30 m depths within 0.50 m intervals along the soil transect to compare soil resistivity with root mass density (RMD. RMD data, up to this depth, varied from 0.000019 to 0.009469 Mg·m−3, showing high spatial variability and significant relationship to the observed values of soil resistivity. These preliminary results showed that the electrical resistivity tomography can contribute to root biomass studies in coffee plants; however, more experiments are necessary to confirm the found results in Brazil coffee plantations.

Chimeric proteins for detection and quantitation of DNA mutations, DNA sequence variations, DNA damage and DNA mismatches

Science.gov (United States)

McCutchen-Maloney, Sandra L.

2002-01-01

Chimeric proteins having both DNA mutation binding activity and nuclease activity are synthesized by recombinant technology. The proteins are of the general formula A-L-B and B-L-A where A is a peptide having DNA mutation binding activity, L is a linker and B is a peptide having nuclease activity. The chimeric proteins are useful for detection and identification of DNA sequence variations including DNA mutations (including DNA damage and mismatches) by binding to the DNA mutation and cutting the DNA once the DNA mutation is detected.

Spin transport, magnetoresistance, and electrically detected magnetic resonance in amorphous hydrogenated silicon nitride

Energy Technology Data Exchange (ETDEWEB)

Mutch, Michael J. [Intercollege Program of Materials, Pennsylvania State University, University Park, Pennsylvania 16802 (United States); Lenahan, Patrick M. [Intercollege Program of Materials, Pennsylvania State University, University Park, Pennsylvania 16802 (United States); Department of Engineering Science and Mechanics, Pennsylvania State University, University Park, Pennsylvania 16802 (United States); King, Sean W. [Logic Technology Development, Intel Corporation, Hillsboro, Oregon 97124 (United States)

2016-08-08

We report on a study of spin transport via electrically detected magnetic resonance (EDMR) and near-zero field magnetoresistance (MR) in silicon nitride films. Silicon nitrides have long been important materials in solid state electronics. Although electronic transport in these materials is not well understood, electron paramagnetic resonance studies have identified a single dominating paramagnetic defect and have also provided physical and chemical descriptions of the defects, called K centers. Our EDMR and MR measurements clearly link the near-zero field MR response to the K centers and also indicate that K center energy levels are approximately 3.1 eV above the a-SiN:H valence band edge. In addition, our results suggest an approach for the study of defect mediated spin-transport in inorganic amorphous insulators via variable electric field and variable frequency EDMR and MR which may be widely applicable.

Electrically-detected electron paramagnetic resonance of point centers in 6H-SiC nanostructures

Czech Academy of Sciences Publication Activity Database

Bagraev, N.T.; Gets, D.S.; Kalabukhova, E.N.; Klyachkin, L.E.; Malyarenko, A.M.; Mashkov, V.A.; Savchenko, Dariia; Shanina, B.D.

2014-01-01

Roč. 48, č. 11 (2014), s. 1467-1480 ISSN 1063-7826 R&D Projects: GA MŠk(CZ) LM2011029 Grant - others:SAFMAT(XE) CZ.2.16/3.1.00/22132 Institutional support: RVO:68378271 Keywords : electron paramagnetic resonance * electrically- detected electron paramagnetic resonance * 6H -SiC nanostructures * nitrogen-vacancy defect * point defect Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 0.739, year: 2014

Analysis of gate underlap channel double gate MOS transistor for electrical detection of bio-molecules

Science.gov (United States)

Ajay; Narang, Rakhi; Saxena, Manoj; Gupta, Mridula

2015-12-01

In this paper, an analytical model for gate drain underlap channel Double-Gate Metal-Oxide-Semiconductor Field-Effect Transistor (DG-MOSFET) for label free electrical detection of biomolecules has been proposed. The conformal mapping technique has been used to derive the expressions for surface potential, lateral electric field, energy bands (i.e. conduction and valence band) and threshold voltage (Vth). Subsequently a full drain current model to analyze the sensitivity of the biosensor has been developed. The shift in the threshold voltage and drain current (after the biomolecules interaction with the gate underlap channel region of the MOS transistor) has been used as a sensing metric. All the characteristic trends have been verified through ATLAS (SILVACO) device simulation results.

Evaluating the efficacy of a structure-derived amino acid substitution matrix in detecting protein homologs by BLAST and PSI-BLAST

Directory of Open Access Journals (Sweden)

Nalin CW Goonesekere

2009-06-01

Full Text Available Nalin CW GoonesekereDepartment of Chemistry and Biochemistry, University of Northern iowa, Cedar Falls, IA, USAAbstract: The large numbers of protein sequences generated by whole genome sequencing projects require rapid and accurate methods of annotation. The detection of homology through computational sequence analysis is a powerful tool in determining the complex evolutionary and functional relationships that exist between proteins. Homology search algorithms employ amino acid substitution matrices to detect similarity between proteins sequences. The substitution matrices in common use today are constructed using sequences aligned without reference to protein structure. Here we present amino acid substitution matrices constructed from the alignment of a large number of protein domain structures from the structural classification of proteins (SCOP database. We show that when incorporated into the homology search algorithms BLAST and PSI-blaST, the structure-based substitution matrices enhance the efficacy of detecting remote homologs. Keywords: computational biology, protein homology, amino acid substitution matrix, protein structure

Protein electrochemistry using graphene-based nano-assembly: an ultrasensitive electrochemical detection of protein molecules via nanoparticle-electrode collisions.

Science.gov (United States)

Li, Da; Liu, Jingquan; Barrow, Colin J; Yang, Wenrong

2014-08-04

We describe a new electrochemical detection approach towards single protein molecules (microperoxidase-11, MP-11), which are attached to the surface of graphene nanosheets. The non-covalently functionalized graphene nanosheets exhibit enhanced electroactive surface area, where amplified redox current is produced when graphene nanosheets collide with the electrode.

Development and application of a fluorescence protein microarray for detecting serum alpha-fetoprotein in patients with hepatocellular carcinoma.

Science.gov (United States)

Zhang, Aiying; Yin, Chengzeng; Wang, Zhenshun; Zhang, Yonghong; Zhao, Yuanshun; Li, Ang; Sun, Huanqin; Lin, Dongdong; Li, Ning

2016-12-01

Objective To develop a simple, effective, time-saving and low-cost fluorescence protein microarray method for detecting serum alpha-fetoprotein (AFP) in patients with hepatocellular carcinoma (HCC). Method Non-contact piezoelectric print techniques were applied to fluorescence protein microarray to reduce the cost of prey antibody. Serum samples from patients with HCC and healthy control subjects were collected and evaluated for the presence of AFP using a novel fluorescence protein microarray. To validate the fluorescence protein microarray, serum samples were tested for AFP using an enzyme-linked immunosorbent assay (ELISA). Results A total of 110 serum samples from patients with HCC ( n = 65) and healthy control subjects ( n = 45) were analysed. When the AFP cut-off value was set at 20 ng/ml, the fluorescence protein microarray had a sensitivity of 91.67% and a specificity of 93.24% for detecting serum AFP. Serum AFP quantified via fluorescence protein microarray had a similar diagnostic performance compared with ELISA in distinguishing patients with HCC from healthy control subjects (area under receiver operating characteristic curve: 0.906 for fluorescence protein microarray; 0.880 for ELISA). Conclusion A fluorescence protein microarray method was developed for detecting serum AFP in patients with HCC.

Detection of HIV-1 and Human Proteins in Urinary Extracellular Vesicles from HIV+ Patients

Directory of Open Access Journals (Sweden)

Samuel I. Anyanwu

2018-01-01

Full Text Available Background. Extracellular vesicles (EVs are membrane bound, secreted by cells, and detected in bodily fluids, including urine, and contain proteins, RNA, and DNA. Our goal was to identify HIV and human proteins (HPs in urinary EVs from HIV+ patients and compare them to HIV− samples. Methods. Urine samples were collected from HIV+ (n=35 and HIV− (n=12 individuals. EVs were isolated by ultrafiltration and characterized using transmission electron microscopy, tandem mass spectrometry (LC/MS/MS, and nanoparticle tracking analysis (NTA. Western blots confirmed the presence of HIV proteins. Gene ontology (GO analysis was performed using FunRich and HIV Human Interaction database (HHID. Results. EVs from urine were 30–400 nm in size. More EVs were in HIV+ patients, P<0.05, by NTA. HIV+ samples had 14,475 HPs using LC/MS/MS, while only 111 were in HIV−. HPs in the EVs were of exosomal origin. LC/MS/MS showed all HIV+ samples contained at least one HIV protein. GO analysis showed differences in proteins between HIV+ and HIV− samples and more than 50% of the published HPs in the HHID interacted with EV HIV proteins. Conclusion. Differences in the proteomic profile of EVs from HIV+ versus HIV− samples were found. HIV and HPs in EVs could be used to detect infection and/or diagnose HIV disease syndromes.

Label-free detection of C-reactive protein using reflectometric interference spectroscopy-based sensing system

Energy Technology Data Exchange (ETDEWEB)

Choi, Hyung Woo; Sakata, Yasuhiko [Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501 (Japan); Kurihara, Yoshikazu [Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501 (Japan); KONICA MINOLTA OPTO, Inc., 1 Sakura-machi, Hino-shi, Tokyo 191-8511 (Japan); Ooya, Tooru [Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501 (Japan); Takeuchi, Toshifumi, E-mail: takeuchi@gold.kobe-u.ac.jp [Graduate School of Engineering, Kobe University, 1-1 Rokkodai-cho, Nada-ku, Kobe 657-8501 (Japan)

2012-05-30

Highlights: Black-Right-Pointing-Pointer A new RIfS-based label-free biosensing system for C-reactive protein was developed. Black-Right-Pointing-Pointer Silicon-based inexpensive chips and the simple optical setup were employed. Black-Right-Pointing-Pointer Owing to the TMS treatment and the use of protein A, the sensitivity was enhanced. Black-Right-Pointing-Pointer It can be applied to other target as a substitute of SPR-based expensive sensors. - Abstract: Reflectometric interference spectroscopy (RIfS) is a label-free, time-resolved technique, and suitable for detecting antibody-antigen interaction. This work describes a continuous flow biosensor for C-reactive protein (CRP), involving an effective immobilization method of a monoclonal antibody against CRP (anti-CRP) to achieve highly sensitive RIfS-based detection of CRP. The silicon nitride-coated silicon chip (SiN chip) for the RIfS sensing was first treated with trimethylsilylchloride (TMS), followed by UV-light irradiation to in situ generation of homogeneous silanols on the surface. Following amination by 3-aminopropyltriethoxysilane, carboxymethyldextran (CMD) was grafted, and subsequently, protein A was immobilized to create the oriented anti-CRP surface. The immobilization process of protein A and anti-CRP was monitored with the RIfS system by consecutive injections of an amine coupling reagent, protein A and anti-CRP, respectively, to confirm the progress of each step in real time. The sensitivity was enhanced when all of the processes were adopted, suggesting that the oriented immobilization of anti-CRP via protein A that was coupled with the grafted CMD on the aminated surface of TMS-treated SiN chip. The feasibility of the present sensing system was demonstrated on the detection of CRP, where the silicon-based inexpensive chips and the simple optical setup were employed. It can be applied to other target molecules in various fields of life science as a substitute of surface plasmon resonance

Use of technical biochemical in combination for the detection of proteins of union to calcium in Plasmodium falciparum

International Nuclear Information System (INIS)

Cabrera, Rodrigo; Wasserman, Moises

2003-01-01

Calcium plays a fundamental role in the development of Plasmodium falciparum, the intracellular parasite that causes malaria. With the purpose of understanding the mechanism by which calcium acts in this parasite, calcium-binding proteins were detected in this organism the combined use of the metachromatic dye Stains-all and the 4 5 C a overlay assay allowed the identification, in mature parasites. Of 9 calcium - binding proteins. 6 of which seem to be different from any reported calcium-binding protein. Additionally, it was determined that the combined use of these techniques can be useful for the detection and purification of calcium-binding proteins

A case study to detect the leakage of underground pressureless cement sewage water pipe using GPR, electrical, and chemical data.

Science.gov (United States)

Liu, Guanqun; Jia, Yonggang; Liu, Hongjun; Qiu, Hanxue; Qiu, Dongling; Shan, Hongxian

2002-03-01

The exploration and determination of leakage of underground pressureless nonmetallic pipes is difficult to deal with. A comprehensive method combining Ground Penetrating Rader (GPR), electric potential survey and geochemical survey is introduced in the leakage detection of an underground pressureless nonmetallic sewage pipe in this paper. Theoretically, in the influencing scope of a leakage spot, the obvious changes of the electromagnetic properties and the physical-chemical properties of the underground media will be reflected as anomalies in GPR and electrical survey plots. The advantages of GPR and electrical survey are fast and accurate in detection of anomaly scope. In-situ analysis of the geophysical surveys can guide the geochemical survey. Then water and soil sampling and analyzing can be the evidence for judging the anomaly is caused by pipe leakage or not. On the basis of previous tests and practical surveys, the GPR waveforms, electric potential curves, contour maps, and chemical survey results are all classified into three types according to the extent or indexes of anomalies in orderto find out the leakage spots. When three survey methods all show their anomalies as type I in an anomalous spot, this spot is suspected as the most possible leakage location. Otherwise, it will be down grade suspected point. The suspect leakage spots should be confirmed by referring the site conditions because some anomalies are caused other factors. The excavation afterward proved that the method for determining the suspected location by anomaly type is effective and economic. Comprehensive method of GRP, electric potential survey, and geochemical survey is one of the effective methods in the leakage detection of underground nonmetallic pressureless pipe with its advantages of being fast and accurate.

Detection of hyperphosphorylated tau protein and α-synuclein in spinal cord of patients with Alzheimer’s disease

Directory of Open Access Journals (Sweden)

Guo YJ

2016-02-01

Full Text Available Yanjun Guo,1,2 Luning Wang,2 Mingwei Zhu,2 Honghong Zhang,3 Yazhuo Hu,3 Zhitao Han,3 Jia Liu,4 Weiqin Zhao,1 Dexin Wang11Department of Neurology, Beijing Friendship Hospital, Capital Medical University, 2Department of Geriatric Neurology, PLA General Hospital, 3Institute of Geriatrics, Chinese PLA General Hospital & Chinese PLA Medical Academy, 4Department of Neurology, Xuanwu Hospital, Capital Medical University, Beijing, People’s Republic of ChinaAbstract: The aim of this study was to investigate the neuropathological features of the spinal cord in patients suffering with Alzheimer’s disease (AD. Spinal cord tissue collected from three AD patients and eight controls was selected for the study. Data were collected at T2, T8, T10, L4, and S2 spinal levels. The sections were subjected to hematoxylin and eosin and Gallyas–Braak staining methods and then were immunostained with antibodies such as phosphorylated tau protein (AT8, α-synuclein, Aβ, amyloid precursor protein , ubiquitin, and TDP-43. Pathological changes exhibited by the biomarkers were detected by microscopy. Neurofibrillary tangles (NFTs were detectable in spinal anterior horn motor neurons in two of the three AD patients. AT8-positive axons or axon-like structures and AT8 expression in glial cells were detected in all three AD cases. Hyperphosphorylation of tau protein was detected in spinal anterior horn cells, glial cells, and axons, and its severity was associated with NFTs in the brain tissue. α-Synuclein-positive Lewy bodies and scattered Lewy-like neuritis were detected in the medial horn of the thoracic spinal cord and ventral sacral gray matter, respectively, in one patient who had AD with Lewy bodies. Neither amyloid deposition nor amyloid precursor protein and TDP-43 expression was detected in the spinal cord of AD patients. Spinal cord of AD patients was observed to contain phosphorylated tau protein and α-synuclein immunoreactive structures, which may play a

Effect of vitamin D status on chick kidney proteins: detection of a 45-kilodalton mitochondrial protein suppressed by vitamin D

International Nuclear Information System (INIS)

Kain, S.R.; Kamrath, K.S.; Henry, H.L.

1988-01-01

Two-dimensional polyacrylamide gel electrophoresis along with L-[ 35 S]methionine radiolabeling studies were used to examine the effect of chronic vitamin D status on the composition and relative abundance of chick kidney proteins. Comparison of silver-stained gels revealed no extensive differences in either the electrophoretic mobility or the amounts of kidney proteins present in the mitochondrial fraction from vitamin D-replete and vitamin D-deficient chicks. A similar result was obtained in studies with L-[ 35 S]methionine-labeled proteins. Vitamin D deficiency specifically elevated levels of a 45-kilodalton mitochondrial protein (pI 5.0 to 5.5) by approximately 5- to 12-fold relative to amounts present in vitamin D-replete tissue. This protein could not be detected in postmitochondrial supernatant fractions and was only faintly visible in crude kidney homogenates. The specificity of the observed suppression of this 45-kilodalton protein by vitamin D suggests that it may play an important role in renal functions influenced by the vitamin D endocrine system

Detection of protein concentrations using a pH-step titration method

NARCIS (Netherlands)

Kruise, J.; Kruise, J.; Eijkel, Jan C.T.; Bergveld, Piet

1997-01-01

A stimulus-response method based on the application of a pH step is proposed for the detection of protein immobilized in a membrane on top of an ion-sensitive field-effect transistor (ISFET). The ISFET response to a step-wise change in pH, applied at the interface between the membrane and the

Electrical detection of hyperfine interactions in silicon

Energy Technology Data Exchange (ETDEWEB)

Hoehne, Felix

2012-12-15

The main focus of this work was the measurement of hyperfine interactions of defects in silicon using EDMR. We combined the high sensitivity of EDMR when compared to conventional ESR with the two most commonly used methods for the measurement of hyperfine interactions: ESEEM and ENDOR. We first demonstrated the electrical detection of ESEEM by measuring the hyperfine interactions of {sup 31}P donors in Si:P with {sup 29}Si nuclear spins. We then apply EDESEEM to P{sub b0} defects at the Si/SiO{sub 2} interface. In isotopically engineered, we observe an ESEEM modulation with a characteristic beating caused by {sup 29}Si nuclei at 4th and 5th nearest neighbor lattice sites. Then we combine pulsed ENDOR with the high sensitivity of EDMR (EDENDOR). First we demonstrate the measurement of {sup 31}P nuclear spin hyperfine transitions and the coherent manipulation and readout of the {sup 31}P nuclear spins under continuous illumination with above bandgap light. We further show that the EDENDOR method can be greatly improved by switching off the illumination during the microwave and rf pulses. This improves the signal-to-noise ratio by two orders of magnitude and removes the non-resonant background induced by the strong rf pulse allowing to measure ENDOR with a sensitivity <3000 nuclear spins. We apply EDENDOR to the {sup 31}P-P{sub b0} spin system and the {sup 31}P-SL1 spin system allowing us to compare the hyperfine interactions of bulk and interface-near donors. The pulsed illumination also makes spectroscopy of the {sup 31}P{sup +} nuclear spin possible, which due to its long coherence time of 18 ms compared to 280 {mu}s for the {sup 31}P{sub 0} nuclear spin, might be a candidate for a nuclear spin memory. In the last part, we devise a scheme for the hyperpolarization of {sup 31}P nuclei by combining pulsed optical excitation and pulsed ENDOR and demonstrate a {sup 31}P nuclear spin polarization of more than 50%. Crucial for these experiments was the development of a

Electrical detection of hyperfine interactions in silicon

International Nuclear Information System (INIS)

Hoehne, Felix

2012-01-01

The main focus of this work was the measurement of hyperfine interactions of defects in silicon using EDMR. We combined the high sensitivity of EDMR when compared to conventional ESR with the two most commonly used methods for the measurement of hyperfine interactions: ESEEM and ENDOR. We first demonstrated the electrical detection of ESEEM by measuring the hyperfine interactions of 31 P donors in Si:P with 29 Si nuclear spins. We then apply EDESEEM to P b0 defects at the Si/SiO 2 interface. In isotopically engineered, we observe an ESEEM modulation with a characteristic beating caused by 29 Si nuclei at 4th and 5th nearest neighbor lattice sites. Then we combine pulsed ENDOR with the high sensitivity of EDMR (EDENDOR). First we demonstrate the measurement of 31 P nuclear spin hyperfine transitions and the coherent manipulation and readout of the 31 P nuclear spins under continuous illumination with above bandgap light. We further show that the EDENDOR method can be greatly improved by switching off the illumination during the microwave and rf pulses. This improves the signal-to-noise ratio by two orders of magnitude and removes the non-resonant background induced by the strong rf pulse allowing to measure ENDOR with a sensitivity 31 P-P b0 spin system and the 31 P-SL1 spin system allowing us to compare the hyperfine interactions of bulk and interface-near donors. The pulsed illumination also makes spectroscopy of the 31 P + nuclear spin possible, which due to its long coherence time of 18 ms compared to 280 μs for the 31 P 0 nuclear spin, might be a candidate for a nuclear spin memory. In the last part, we devise a scheme for the hyperpolarization of 31 P nuclei by combining pulsed optical excitation and pulsed ENDOR and demonstrate a 31 P nuclear spin polarization of more than 50%. Crucial for these experiments was the development of a lock-in detection scheme for pEDMR, which improves the signal-to-noise ratio by one order of magnitude by removing low

Protein A磁珠检测朊病毒方法的建立%The foundation of Protein A magnetism to detect Prion assay

Institute of Scientific and Technical Information of China (English)

李奇; 杨阳; 吕鹏; 宋博翠; 白宗瑞; 陈志宝

2009-01-01

建立Protein A磁珠法检测朊病毒,将朊病毒抗体IH2包被后的Protein A磁珠与蛋白酶K消化过的攻毒小鼠脑组织匀浆液混合,经免疫沉淀缓冲液温和洗脱杂蛋白,再经0.2 M甘氨酸将目的蛋白洗脱并且利用聚丙酰胺凝胶电泳检测.通过聚丙酰胺凝胶电泳检测出25 KD的目的蛋白带,经与正常未消化的朊蛋白条带相比较,证实检测出朊病毒.Protein A磁珠具有高效特异的与抗体IgG结合的特点,与多克隆抗体混合后可有效结合IgG.该方法较免疫组织化学和Western-blot技术简便快速,且磁珠可以回收再使用,使成本低廉.%Objective The country need the fast and effective detection for Prion asthe high risk country where Scrapie and BSE may bresk out. The common assay of Western-Blot and Immunohistochemistry consume a long time and the associated experience is needed. Protein A magnetism has the feature of binding IgG differential.h could bind the IgG adequately,when mixing with polyclonal antibody.After intermixing with brain tissue protein digested by protein kinase, Prion could easily elute from Protein A magnetism.Interest protein band is 25KD through SDS-PAGE. When definiting the molecular weight of Priori,the result of detection is educed by SDS-PAGE electrophoresis. The assay is more handier and faster than common assay. The magnetism can reclaim and reuse,which reduce the cost.The assay of detection is a innovation and many branches could generally applicated it.

Development of ELISA for the detection of transgenic vegetative insecticidal protein in GM crops/produce.

Science.gov (United States)

Kumar, R

2012-01-11

In the process of the development of insect-resistant genetically modified (GM) crops and also to evaluate the consistency in the expression of toxin under field conditions, immunological assays are commonly being used. An immunoassay was developed to support the labelling of vegetative insecticidal protein (Vip3A)-based GM produce. The developed ELISA for the measurement of Vip3A is a triple antibody sandwich procedure utilising a polyclonal capture antibody (mouse anti-Vip3A) and a polyclonal detection antibody (rabbit anti-Vip3A) followed by use of a third HRP-conjugated anti-species antibody (goat anti-rabbit IgG). The limit of detection limit of the ELISA assay was 16 ng ml(-1) with a linear quantification range from approximately 31 to 500 ng ml(-1) of Vip3A protein. Furthermore, the assay was in-house validated with GM brinjal samples. The assay was specific, sensitive and reproducible, which can be helpful to detect and track down the spread of unapproved and intentionally/unintentionally released GM produce harbouring Vip protein.

Atmospheric electric field effects of cosmic rays detected in Mexico City

Energy Technology Data Exchange (ETDEWEB)

Gonzalez, L. X; Valdes-Galicia, J. F [Instituto de Geofisica, Universidad National Autonoma de Mexico, Mexico D.F(Mexico)

2006-10-15

We studied the possible effects of atmospheric electric fields, generated in thunderstorms, on the cosmic ray intensity detected at the Earth's surface by investigating the variations of the counting rates of the cosmic-ray nucleonic component, obtained from the neutron monitor installed in Mexico City, for thunderstorms during 1996 and 1997. These were years of minimum solar activity. We compare our experimental results with the general theory of cosmic ray meteorological effects by Dorman (1995). The observed intensity variation is about 0.2%. According to Dorman (1995), the effect should be between 0.27% and 0.81% on the counting rate of the neutron monitor when the atmospheric electric field intensities are around 100 to 300 Vcm-1.Our results show that either the electric field in Mexico City had less intensity than assumed by Dorman (1995), or the electric field is not uniform in time and height during the development of the thunderstorm. [Spanish] Estudiamos los posibles efectos de los campos electricos atmosfericos, generados en las tormentas electricas, sobre la intensidad de los rayos cosmicos detectados en la superficie terrestre, analizando las variaciones de las razones de conteo de la componente nucleonica de los rayos cosmicos, obtenidas por el monitor de neutrones instalado en la ciudad de Mexico, durante tormentas electricas ocurridas entre 1996 y 1997, anos del minimo solar. Comparamos nuestros resultados experimentales con la teoria general de los efectos meteorologicos en los rayos cosmicos, desarrollada por Dorman (1995). Se observo una variacion en la intensidad de alrededor de 0.2%. De acuerdo con Dorman (1995), el efecto puede estar entre 0.27 % y 0.81% en las razones de conteo del monitor de neutrones cuando las intensidades del campo electrico atmosferico se encuentran al rededor de 100 a 300 Vcm-1. Nuestros resultados muestran que los campos electricos en la ciudad de Mexico tuvieron menos intensidad que los campos electricos asumidos

Detection of high molecular weight proteins by MALDI imaging mass spectrometry.

Science.gov (United States)

Mainini, Veronica; Bovo, Giorgio; Chinello, Clizia; Gianazza, Erica; Grasso, Marco; Cattoretti, Giorgio; Magni, Fulvio

2013-06-01

MALDI imaging mass spectrometry (IMS) is a unique technology to explore the spatial distribution of biomolecules directly on tissues. It allows the in situ investigation of a large number of small proteins and peptides. Detection of high molecular weight proteins through MALDI IMS still represents an important challenge, as it would allow the direct investigation of the distribution of more proteins involved in biological processes, such as cytokines, enzymes, neuropeptide precursors and receptors. In this work we compare the traditional method performed with sinapinic acid with a comparable protocol using ferulic acid as the matrix. Data show a remarkable increase of signal acquisition in the mass range of 20k to 150k Th. Moreover, we report molecular images of biomolecules above 70k Th, demonstrating the possibility of expanding the application of this technology both in clinical investigations and basic science.

Sensitive electrochemiluminescence biosensor based on Au-ITO hybrid bipolar electrode amplification system for cell surface protein detection.

Science.gov (United States)

Wu, Mei-Sheng; Yuan, Da-Jing; Xu, Jing-Juan; Chen, Hong-Yuan

2013-12-17

Here we developed a novel hybrid bipolar electrode (BPE)-electrochemiluminescence (ECL) biosensor based on hybrid bipolar electrode (BPE) for the measurement of cancer cell surface protein using ferrocence (Fc) labeled aptamer as signal recognition and amplification probe. According to the electric neutrality of BPE, the cathode of U-shaped ITO BPE was electrochemically deposited by Au nanoparticles (NPs) to enhance its conductivity and surface area, decrease the overpotential of O2 reduction, which would correspondingly increase the oxidation current of Ru(bpy)3(2+)/tripropylamine (TPA) on the anode of BPE and resulting a ∼4-fold enhancement of ECL intensity. Then a signal amplification strategy was designed by introducing Fc modified aptamer on the anode surface of BPE through hybridization for detecting the amount of mucin-1 on MCF-7 cells. The presence of Fc could not only inhibit the oxidation of Ru(bpy)3(2+) because of its lower oxidation potential, its oxidation product Fc(+) could also quench the ECL of Ru(bpy)3(2+)/TPA by efficient energy-transfer from the excited-state Ru(bpy)3(2+)* to Fc(+), making the ECL intensity greatly quenched. On the basis of the cathodic Au NPs induced ECL enhancing coupled with anodic Fc induced signal quenching amplification, the approach allowed detection of mucin-1 aptamer at a concentration down to 0.5 fM and was capable of detecting a minimum of 20 MCF-7 cells. Besides, the amount of mucin-1 on MCF-7 cells was calculated to be 9041 ± 388 molecules/cell. This approach therefore shows great promise in bioanalysis.

A feasibility study of magnetic resonance electrical impedance tomography for prostate cancer detection

International Nuclear Information System (INIS)

Liu, Yang; Zhang, Yingchun

2014-01-01

Magnetic resonance electrical impedance tomography (MREIT) is an imaging technique that reconstructs the conductivity distribution inside the subject using magnetic flux density or current density measurements acquired by a magnetic resonance imaging system. Since the primary prostate cancer diagnostic method, prostate biopsy, has limited accuracy in cancer diagnosis and malignant tissues have shown significantly different electrical properties from normal or benign tissues, MREIT has potential application in prostate cancer detection. The feasibility of utilizing MREIT in detecting prostate cancer was evaluated via a series of well-designed computer simulations in the present study. MREIT techniques with three different electrode configurations (external, trans-rectal, and trans-urethral electrode arrays) and two different reconstruction algorithms (J-substitution algorithm and harmonic B z  algorithm) were successfully developed. The performance of different MREIT techniques were evaluated and compared based on the imaging accuracy of the reconstructed conductivity distribution in the prostate. Without the presence of noise, the external MREIT achieves a better imaging accuracy than the two endo-MREIT (trans-rectal and trans-urethral) techniques, while the trans-urethral MREIT achieves the best imaging accuracy in noisy environments. We also found that the J-substitution reconstruction algorithm consistently offered better imaging accuracy than the harmonic B z  algorithm. When Gaussian distributed random noise with a standard deviation of 0.25 nT was added, the relative errors (RE) between the reconstructed and target conductivity distributions inside the prostate were observed to be 14.18% and 17.35% by the trans-urethral MREIT with the J-substitution and harmonic B z  algorithms respectively. The lower REs of 9.64% and 11.17% were achieved respectively when the standard deviation of noise was reduced to 0.05 nT. The simulation results demonstrate the

Detection of protein-small molecule binding using a self-referencing external cavity laser biosensor.

Science.gov (United States)

Meng Zhang; Peh, Jessie; Hergenrother, Paul J; Cunningham, Brian T

2014-01-01

High throughput screening of protein-small molecule binding interactions using label-free optical biosensors is challenging, as the detected signals are often similar in magnitude to experimental noise. Here, we describe a novel self-referencing external cavity laser (ECL) biosensor approach that achieves high resolution and high sensitivity, while eliminating thermal noise with sub-picometer wavelength accuracy. Using the self-referencing ECL biosensor, we demonstrate detection of binding between small molecules and a variety of immobilized protein targets with binding affinities or inhibition constants in the sub-nanomolar to low micromolar range. The demonstrated ability to perform detection in the presence of several interfering compounds opens the potential for increasing the throughput of the approach. As an example application, we performed a "needle-in-the-haystack" screen for inhibitors against carbonic anhydrase isozyme II (CA II), in which known inhibitors are clearly differentiated from inactive molecules within a compound library.

Detection of protein-protein interactions by ribosome display and protein in situ immobilisation.

Science.gov (United States)

He, Mingyue; Liu, Hong; Turner, Martin; Taussig, Michael J

2009-12-31

We describe a method for identification of protein-protein interactions by combining two cell-free protein technologies, namely ribosome display and protein in situ immobilisation. The method requires only PCR fragments as the starting material, the target proteins being made through cell-free protein synthesis, either associated with their encoding mRNA as ribosome complexes or immobilised on a solid surface. The use of ribosome complexes allows identification of interacting protein partners from their attached coding mRNA. To demonstrate the procedures, we have employed the lymphocyte signalling proteins Vav1 and Grb2 and confirmed the interaction between Grb2 and the N-terminal SH3 domain of Vav1. The method has promise for library screening of pairwise protein interactions, down to the analytical level of individual domain or motif mapping.

Electrochemical platform for the detection of transmembrane proteins reconstituted into liposomes

Czech Academy of Sciences Publication Activity Database

Vacek, J.; Zatloukalová, M.; Geletičová, J.; Kubala, M.; Modriansky, M.; Fekete, Ladislav; Mašek, J.; Hubatka, F.; Turánek, J.

2016-01-01

Roč. 88, č. 8 (2016), s. 4548-4556 ISSN 0003-2700 R&D Projects: GA MŠk LO1409; GA MŠk LM2015088 Institutional support: RVO:68378271 Keywords : detection * transmembrane proteins * liposomes * electrochemistry Subject RIV: BM - Solid Matter Physics ; Magnetism OBOR OECD: Condensed matter physics (including formerly solid state physics, supercond.) Impact factor: 6.320, year: 2016

A novel approach for the detection of potentially hazardous pepsin stable hazelnut proteins as contaminants in chocolate-based food.

Science.gov (United States)

Akkerdaas, Jaap H; Wensing, Marjolein; Knulst, André C; Stephan, Oliver; Hefle, Susan L; Aalberse, Rob C; van Ree, Ronald

2004-12-15

Contamination of food products with pepsin resistant allergens is generally believed to be a serious threat to patients with severe food allergy. A sandwich type enzyme-linked immunosorbent assay (ELISA) was developed to measure pepsin resistant hazelnut protein in food products. Capturing and detecting rabbit antibodies were raised against pepsin-digested hazelnut and untreated hazelnut protein, respectively. The assay showed a detection limit of 0.7 ng/mL hazelnut protein or food matrix and a maximum of 0.034% cross-reactivity (peanut). Chocolate samples spiked with 0.5-100 microg hazelnut/g chocolate showed a mean recovery of 97.3%. In 9/12 food products labeled "may contain nuts", hazelnut was detected between 1.2 and 417 microg hazelnut/g food. It can be concluded that the application of antibodies directed to pepsin-digested food extracts in ELISA can facilitate specific detection of stable proteins that have the highest potential of inducing severe food anaphylaxis.

A pilot study to evaluate the application of a generic protein standard panel for quality control of biomarker detection technologies

Directory of Open Access Journals (Sweden)

Valdivia Hernan J

2011-08-01

Full Text Available Abstract Background Protein biomarker studies are currently hampered by a lack of measurement standards to demonstrate quality, reliability and comparability across multiple assay platforms. This is especially pertinent for immunoassays where multiple formats for detecting target analytes are commonly used. Findings In this pilot study a generic panel of six non-human protein standards (50 - 10^7 pg/mL of varying abundance was prepared as a quality control (QC material. Simulated "normal" and "diseased" panels of proteins were prepared in pooled human plasma and incorporated into immunoassays using the Meso Scale Discovery® (MSD® platform to illustrate reliable detection of the component proteins. The protein panel was also evaluated as a spike-in material for a model immunoassay involving detection of ovarian cancer biomarkers within individual human plasma samples. Our selected platform could discriminate between two panels of the proteins exhibiting small differences in abundance. Across distinct experiments, all component proteins exhibited reproducible signal outputs in pooled human plasma. When individual donor samples were used, half the proteins produced signals independent of matrix effects. These proteins may serve as a generic indicator of platform reliability. Each of the remaining proteins exhibit differential signals across the distinct samples, indicative of sample matrix effects, with the three proteins following the same trend. This subset of proteins may be useful for characterising the degree of matrix effects associated with the sample which may impact on the reliability of quantifying target diagnostic biomarkers. Conclusions We have demonstrated the potential utility of this panel of standards to act as a generic QC tool for evaluating the reproducibility of the platform for protein biomarker detection independent of serum matrix effects.

Electrical machines diagnosis

CERN Document Server

Trigeassou, Jean-Claude

2013-01-01

Monitoring and diagnosis of electrical machine faults is a scientific and economic issue which is motivated by objectives for reliability and serviceability in electrical drives.This book provides a survey of the techniques used to detect the faults occurring in electrical drives: electrical, thermal and mechanical faults of the electrical machine, faults of the static converter and faults of the energy storage unit.Diagnosis of faults occurring in electrical drives is an essential part of a global monitoring system used to improve reliability and serviceability. This diagnosis is perf

Organic conjugated small molecule materials based optical probe for rapid, colorimetric and UV-vis spectral detection of phosphorylated protein in placental tissue.

Science.gov (United States)

Wang, Yanfang; Yang, Na; Liu, Yi

2018-04-05

A novel organic small molecule with D-Pi-A structure was prepared, which was found to be a promising colorimetric and ratiometric UV-vis spetral probe for detection of phosphorylated proteins with the help of tetravalent zirconium ion. Such optical probe based on chromophore WYF-1 shows a rapid response (within 10s) and high selectivity and sensitivity for phosphorylated proteins, giving distinct colorimetric and ratiometric UV-vis changes at 720 and 560nm. The detection limit for phosphorylated proteins was estimated to be 100nM. In addition, detection of phosphorylated proteins in placental tissue samples with this probe was successfully applied, which indicates that this probe holds great potential for phosphorylated proteins detection. Copyright © 2018 Elsevier B.V. All rights reserved.

Using an alternate light source to detect electrically singed feathers and hair in a forensic setting.

Science.gov (United States)

Viner, Tabitha C; Kagan, Rebecca A; Johnson, Jennifer L

2014-01-01

Mortality due to electrical injury in wildlife may occur in the form of lightning strike or power line contact. Evidence of electrical contact may be grossly obvious, with extensive singeing, curling, and blackening of feathers, fur, or skin. Occasionally, changes may be subtle, owing to lower current or reduced conductivity, making a definitive diagnosis of electrocution more difficult. We describe the use of an alternate light source in the examination of cases of lightning strike and power line contact in wildlife, and the enhanced detection of changes due to electrical currents in the hair and feathers of affected animals. Subtle changes in the wing feathers of 12 snow geese and 1 wolf that were struck by separate lightning events were made obvious by the use of an alternate light source. Similarly, this technique can be used to strengthen the evidence for power line exposure in birds. Published by Elsevier Ireland Ltd.

Novel Phospholipid-Protein Conjugates Allow Improved Detection of Antibodies in Patients with Autoimmune Diseases

DEFF Research Database (Denmark)

Samuelsen, Simone V; Maity, Arindam; Nybo, Mads

2016-01-01

Reliable measurement of clinically relevant autoimmune antibodies toward phospholipid-protein conjugates is highly desirable in research and clinical assays. To date, the development in this field has been limited to the use of natural heterogeneous antigens. However, this approach does not take ...... on the correlation of detected autoantibodies with disease activity and manifestations. This confirms the crucial importance of antigens' composition on research and diagnostic assays, and opens up exciting perspectives for synthetic antigens in future studies of autoimmunity.......Reliable measurement of clinically relevant autoimmune antibodies toward phospholipid-protein conjugates is highly desirable in research and clinical assays. To date, the development in this field has been limited to the use of natural heterogeneous antigens. However, this approach does not take...... structural features of biologically active antigens into account and leads to low reliability and poor scientific test value. Here we describe novel phospholipid-protein conjugates for specific detection of human autoimmune antibodies. Our synthetic approach includes mild oxidation of synthetic phospholipid...

Detection of Foodborne Pathogenic Bacteria using Bacteriophage Tail Spike Proteins

Science.gov (United States)

Poshtiban, Somayyeh

Foodborne infections are worldwide health problem with tremendous social and financial impacts. Efforts are focused on developing accurate and reliable technologies for detection of food contaminations in early stages preferably on-site. This thesis focuses on interfacing engineering and biology by combining phage receptor binding proteins (RBPs) with engineered platforms including microresonator-based biosensors, magnetic particles and polymerase chain reaction (PCR) to develop bacterial detection sensors. We used phage RBPs as target specific bioreceptors to develop an enhanced microresonator array for bacterial detection. These resonator beams are optimized to feature a high natural frequency while offer large surface area for capture of bacteria. Theoretical analysis indicates a high mass sensitivity with a threshold for the detection of a single bacterial cell. We used phage RBPs as target specific bioreceptors, and successfully demonstrated the application of these phage RBB-immobilized arrays for specific detection of C. jejuni cells. We also developed a RBP-derivatized magnetic pre-enrichment method as an upstream sample preparation method to improve sensitivity and specificity of PCR for detection of bacterial cells in various food samples. The combination of RBP-based magnetic separation and real-time PCR allowed the detection of small number of bacteria in artificially contaminated food samples without any need for time consuming pre-enrichment step through culturing. We also looked into integration of the RBP-based magnetic separation with PCR onto a single microfluidic lab-on-a-chip to reduce the overall turnaround time.

An electric detection of immunoglobulin G in the enzyme-linked immunosorbent assay using an indium oxide nanoparticle ion-sensitive field-effect transistor

International Nuclear Information System (INIS)

Lee, Dongjin; Cui, Tianhong

2012-01-01

Semiconducting nanoparticle ion-sensitive field-effect transistors (ISFETs) are used to detect immunoglobulin G (IgG) in the conventional enzyme-linked immunosorbent assay (ELISA). Indium oxide and silica nanoparticles were layer-by-layer self-assembled with the oppositely charged polyelectrolyte as the electrochemical transducer and antibody immobilization site, respectively. The assay was conducted on a novel platform of indium oxide nanoparticle ISFETs, where the electric signals are generated in response to the concentration of target IgG using the labeled detecting antibody. The sandwiched ELISA structure catalyzed the conversion of the acidic substrate into neutral substance with the aid of horseradish peroxidase. The pH change in the substrate solution was detected by nanoparticle ISFETs. Normal rabbit IgG was used as a model antigen whose detection limit of 0.04 ng ml −1 was found. The facile electric detection in the conventional assay through the semiconducting nanoparticle ISFET has potential applications as a point-of-care detection or a sensing element in a lab-on-a-chip system

Detection of secondary structure elements in proteins by hydrophobic cluster analysis.

Science.gov (United States)

Woodcock, S; Mornon, J P; Henrissat, B

1992-10-01

Hydrophobic cluster analysis (HCA) is a protein sequence comparison method based on alpha-helical representations of the sequences where the size, shape and orientation of the clusters of hydrophobic residues are primarily compared. The effectiveness of HCA has been suggested to originate from its potential ability to focus on the residues forming the hydrophobic core of globular proteins. We have addressed the robustness of the bidimensional representation used for HCA in its ability to detect the regular secondary structure elements of proteins. Various parameters have been studied such as those governing cluster size and limits, the hydrophobic residues constituting the clusters as well as the potential shift of the cluster positions with respect to the position of the regular secondary structure elements. The following results have been found to support the alpha-helical bidimensional representation used in HCA: (i) there is a positive correlation (clearly above background noise) between the hydrophobic clusters and the regular secondary structure elements in proteins; (ii) the hydrophobic clusters are centred on the regular secondary structure elements; (iii) the pitch of the helical representation which gives the best correspondence is that of an alpha-helix. The correspondence between hydrophobic clusters and regular secondary structure elements suggests a way to implement variable gap penalties during the automatic alignment of protein sequences.

1H-detected MAS solid-state NMR experiments enable the simultaneous mapping of rigid and dynamic domains of membrane proteins

Science.gov (United States)

Gopinath, T.; Nelson, Sarah E. D.; Veglia, Gianluigi

2017-12-01

Magic angle spinning (MAS) solid-state NMR (ssNMR) spectroscopy is emerging as a unique method for the atomic resolution structure determination of native membrane proteins in lipid bilayers. Although 13C-detected ssNMR experiments continue to play a major role, recent technological developments have made it possible to carry out 1H-detected experiments, boosting both sensitivity and resolution. Here, we describe a new set of 1H-detected hybrid pulse sequences that combine through-bond and through-space correlation elements into single experiments, enabling the simultaneous detection of rigid and dynamic domains of membrane proteins. As proof-of-principle, we applied these new pulse sequences to the membrane protein phospholamban (PLN) reconstituted in lipid bilayers under moderate MAS conditions. The cross-polarization (CP) based elements enabled the detection of the relatively immobile residues of PLN in the transmembrane domain using through-space correlations; whereas the most dynamic region, which is in equilibrium between folded and unfolded states, was mapped by through-bond INEPT-based elements. These new 1H-detected experiments will enable one to detect not only the most populated (ground) states of biomacromolecules, but also sparsely populated high-energy (excited) states for a complete characterization of protein free energy landscapes.

Electric-Field-Induced Superconductivity Detected by Magnetization Measurements of an Electric-Double-Layer Capacitor

NARCIS (Netherlands)

Kasahara, Yuichi; Nishijima, Takahiro; Sato, Tatsuya; Takeuchi, Yuki; Ye, Jianting; Yuan, Hongtao; Shimotani, Hidekazu; Iwasa, Yoshihiro

We report evidence for superconductivity induced by the application of strong electric fields onto the surface of a band insulator, ZrNCl, provided by the observation of a shielding diamagnetic signal. We introduced an electric-double-layer capacitor configuration and in situ magnetization

Detection of trans-cis flips and peptide-plane flips in protein structures

NARCIS (Netherlands)

Touw, W.G.; Joosten, R.P.; Vriend, G.

2015-01-01

A coordinate-based method is presented to detect peptide bonds that need correction either by a peptide-plane flip or by a trans-cis inversion of the peptide bond. When applied to the whole Protein Data Bank, the method predicts 4617 trans-cis flips and many thousands of hitherto unknown

Enhanced production and purification of recombinant surface array protein (Sap) for use in detection of Bacillus anthracis.

Science.gov (United States)

Puranik, Nidhi; Tripathi, N K; Pal, V; Goel, Ajay Kumar

2018-05-01

Surface array protein (Sap) can be an important biomarker for specific detection of Bacillus anthracis , which is released by the bacterium during its growth in culture broth. In the present work, we have cloned and expressed Sap in Escherichia coli . The culture conditions and cultivation media were optimized and used in batch fermentation process for scale up of Sap in soluble form. The recombinant Sap was purified employing affinity chromatography followed by diafiltration. The final yield of purified protein was 20 and 46 mg/l of culture during shake flasks and batch fermentation, respectively. The protein purity and its reactivity were confirmed employing SDS-PAGE and Western blot, respectively. The antibodies raised against purified Sap were evaluated by Western blotting for detection of Sap released by B. anthracis . Our results showed that the Sap could be a novel marker for detection and confirmation of B. anthracis .

A rapid and cost-effective fluorescence detection in tube (FDIT) method to analyze protein phosphorylation.

Science.gov (United States)

Jin, Xiao; Gou, Jin-Ying

2016-01-01

Protein phosphorylation is one of the most important post-translational modifications catalyzed by protein kinases in living organisms. The advance of genome sequencing provided the information of protein kinase families in many organisms, including both model and non-model plants. The development of proteomics technologies also enabled scientists to efficiently reveal a large number of protein phosphorylations of an organism. However, kinases and phosphorylation targets are still to be connected to illustrate the complicated network in life. Here we adapted Pro-Q ® Diamond (Pro-Q ® Diamond Phosphoprotein Gel Stain), a widely used phosphoprotein gel-staining fluorescence dye, to establish a rapid, economical and non-radioactive fluorescence detection in tube (FDIT) method to analyze phosphorylated proteins. Taking advantages of high sensitivity and specificity of Pro-Q ® diamond, the FDIT method is also demonstrated to be rapid and reliable, with a suitable linear range for in vitro protein phosphorylation. A significant and satisfactory protein kinase reaction was detected as fast as 15 min from Wheat Kinase START 1.1 (WKS1.1) on a thylakoid ascorbate peroxidase (tAPX), an established phosphorylation target in our earlier study. The FDIT method saves up to 95% of the dye consumed in a gel staining method. The FDIT method is remarkably quick, highly reproducible, unambiguous and capable to be scaled up to dozens of samples. The FDIT method could serve as a simple and sensitive alternative procedure to determine protein kinase reactions with zero radiation exposure, as a supplementation to other widely used radioactive and in-gel assays.

A rapid and cost-effective fluorescence detection in tube (FDIT method to analyze protein phosphorylation

Directory of Open Access Journals (Sweden)

Xiao Jin

2016-11-01

Full Text Available Abstract Background Protein phosphorylation is one of the most important post-translational modifications catalyzed by protein kinases in living organisms. The advance of genome sequencing provided the information of protein kinase families in many organisms, including both model and non-model plants. The development of proteomics technologies also enabled scientists to efficiently reveal a large number of protein phosphorylations of an organism. However, kinases and phosphorylation targets are still to be connected to illustrate the complicated network in life. Results Here we adapted Pro-Q® Diamond (Pro-Q® Diamond Phosphoprotein Gel Stain, a widely used phosphoprotein gel-staining fluorescence dye, to establish a rapid, economical and non-radioactive fluorescence detection in tube (FDIT method to analyze phosphorylated proteins. Taking advantages of high sensitivity and specificity of Pro-Q® diamond, the FDIT method is also demonstrated to be rapid and reliable, with a suitable linear range for in vitro protein phosphorylation. A significant and satisfactory protein kinase reaction was detected as fast as 15 min from Wheat Kinase START 1.1 (WKS1.1 on a thylakoid ascorbate peroxidase (tAPX, an established phosphorylation target in our earlier study. Conclusion The FDIT method saves up to 95% of the dye consumed in a gel staining method. The FDIT method is remarkably quick, highly reproducible, unambiguous and capable to be scaled up to dozens of samples. The FDIT method could serve as a simple and sensitive alternative procedure to determine protein kinase reactions with zero radiation exposure, as a supplementation to other widely used radioactive and in-gel assays.

Recombinant Protein Production from TPO Gen Cloning and Expression for Early Detection of Autoimmune Thyroid Diseases

Science.gov (United States)

Aulanni'am, Aulanni'am; Kinasih Wuragil, Dyah; Wahono Soeatmadji, Djoko; Zulkarnain; Marhendra, Agung Pramana W.

2018-01-01

Autoimmune Thyroid Disease (AITD) is an autoimmune disease that has many clinical symptoms but is difficult to detect at the onset of disease progression. Most thyroid autoimmune disease patients are positive with high titre of thyroid autoantibodies, especially thyroid peroxidase (TPO). The detection AITD are still needed because these tests are extremely high cost and have not regularly been performed in most of clinical laboratories. In the past, we have explored the autoimmune disease marker and it has been developed as source of polyclonal antibodies from patient origin. In the current study, we develop recombinant protein which resulted from cloning and expression of TPO gene from normal person and AITD patients. This work flows involves: DNA isolation and PCR to obtain TPO gene from human blood, insertion of TPO gene to plasmid and transformation to E. coli BL21, Bacterial culture to obtain protein product, protein purification and product analysis. This products can use for application to immunochromatography based test. This work could achieved with the goal of producing autoimmune markers with a guaranteed quality, sensitive, specific and economically. So with the collaboration with industries these devices could be used for early detection. Keywords: recombinant protein, TPO gene, Autoimmune thyroid diseases (AITD)ction of the diseases in the community.

Detection of IgG antibodies against Bordetella pertussis with 125I-protein A

International Nuclear Information System (INIS)

Wirsing von Koenig, C.H.; Finger, H.

1981-01-01

A method for the detection of IgG antibodies against Bordetella pertussis is described, based on the principle of 'sandwich' radioimmunoassay. 125 I protein A is used as radioactive tracer. The influence of amounts of antigen, antibody, radioactive tracer, incubation time and temperature were tested and the optimal conditions for the assay are described. The procedure offers a simple, quick, and sensitive method for detecting antibodies against B. pertussis. Application and limitation of the test are discussed. (orig.)

Protein-protein interactions: an application of Tus-Ter mediated protein microarray system.

Science.gov (United States)

Sitaraman, Kalavathy; Chatterjee, Deb K

2011-01-01

In this chapter, we present a novel, cost-effective microarray strategy that utilizes expression-ready plasmid DNAs to generate protein arrays on-demand and its use to validate protein-protein interactions. These expression plasmids were constructed in such a way so as to serve a dual purpose of synthesizing the protein of interest as well as capturing the synthesized protein. The microarray system is based on the high affinity binding of Escherichia coli "Tus" protein to "Ter," a 20 bp DNA sequence involved in the regulation of DNA replication. The protein expression is carried out in a cell-free protein synthesis system, with rabbit reticulocyte lysates, and the target proteins are detected either by labeled incorporated tag specific or by gene-specific antibodies. This microarray system has been successfully used for the detection of protein-protein interaction because both the target protein and the query protein can be transcribed and translated simultaneously in the microarray slides. The utility of this system for detecting protein-protein interaction is demonstrated by a few well-known examples: Jun/Fos, FRB/FKBP12, p53/MDM2, and CDK4/p16. In all these cases, the presence of protein complexes resulted in the localization of fluorophores at the specific sites of the immobilized target plasmids. Interestingly, during our interactions studies we also detected a previously unknown interaction between CDK2 and p16. Thus, this Tus-Ter based system of protein microarray can be used for the validation of known protein interactions as well as for identifying new protein-protein interactions. In addition, it can be used to examine and identify targets of nucleic acid-protein, ligand-receptor, enzyme-substrate, and drug-protein interactions.

Detection of Protein Interactions in T3S Systems Using Yeast Two-Hybrid Analysis.

Science.gov (United States)

Nilles, Matthew L

2017-01-01

Two-hybrid systems, sometimes termed interaction traps, are genetic systems designed to find and analyze interactions between proteins. The most common systems are yeast based (commonly Saccharomyces cerevisae) and rely on the functional reconstitution of the GAL4 transcriptional activator. Reporter genes, such as the lacZ gene of Escherichia coli (encodes β-galactosidase), are placed under GAL4-dependent transcriptional control to provide quick and reliable detection of protein interactions. In this method the use of a yeast-based two-hybrid system is described to study protein interactions between components of type III secretion systems.

Orthogonal Electric Field Measurements near the Green Fluorescent Protein Fluorophore through Stark Effect Spectroscopy and pKa Shifts Provide a Unique Benchmark for Electrostatics Models.

Science.gov (United States)

Slocum, Joshua D; First, Jeremy T; Webb, Lauren J

2017-07-20

Measurement of the magnitude, direction, and functional importance of electric fields in biomolecules has been a long-standing experimental challenge. pK a shifts of titratable residues have been the most widely implemented measurements of the local electrostatic environment around the labile proton, and experimental data sets of pK a shifts in a variety of systems have been used to test and refine computational prediction capabilities of protein electrostatic fields. A more direct and increasingly popular technique to measure electric fields in proteins is Stark effect spectroscopy, where the change in absorption energy of a chromophore relative to a reference state is related to the change in electric field felt by the chromophore. While there are merits to both of these methods and they are both reporters of local electrostatic environment, they are fundamentally different measurements, and to our knowledge there has been no direct comparison of these two approaches in a single protein. We have recently demonstrated that green fluorescent protein (GFP) is an ideal model system for measuring changes in electric fields in a protein interior caused by amino acid mutations using both electronic and vibrational Stark effect chromophores. Here we report the changes in pK a of the GFP fluorophore in response to the same mutations and show that they are in excellent agreement with Stark effect measurements. This agreement in the results of orthogonal experiments reinforces our confidence in the experimental results of both Stark effect and pK a measurements and provides an excellent target data set to benchmark diverse protein electrostatics calculations. We used this experimental data set to test the pK a prediction ability of the adaptive Poisson-Boltzmann solver (APBS) and found that a simple continuum dielectric model of the GFP interior is insufficient to accurately capture the measured pK a and Stark effect shifts. We discuss some of the limitations of this

Detecting mutually exclusive interactions in protein-protein interaction maps.

KAUST Repository

Sánchez Claros, Carmen

2012-06-08

Comprehensive protein interaction maps can complement genetic and biochemical experiments and allow the formulation of new hypotheses to be tested in the system of interest. The computational analysis of the maps may help to focus on interesting cases and thereby to appropriately prioritize the validation experiments. We show here that, by automatically comparing and analyzing structurally similar regions of proteins of known structure interacting with a common partner, it is possible to identify mutually exclusive interactions present in the maps with a sensitivity of 70% and a specificity higher than 85% and that, in about three fourth of the correctly identified complexes, we also correctly recognize at least one residue (five on average) belonging to the interaction interface. Given the present and continuously increasing number of proteins of known structure, the requirement of the knowledge of the structure of the interacting proteins does not substantially impact on the coverage of our strategy that can be estimated to be around 25%. We also introduce here the Estrella server that embodies this strategy, is designed for users interested in validating specific hypotheses about the functional role of a protein-protein interaction and it also allows access to pre-computed data for seven organisms.

Detecting mutually exclusive interactions in protein-protein interaction maps.

KAUST Repository

Sá nchez Claros, Carmen; Tramontano, Anna

2012-01-01

Comprehensive protein interaction maps can complement genetic and biochemical experiments and allow the formulation of new hypotheses to be tested in the system of interest. The computational analysis of the maps may help to focus on interesting cases and thereby to appropriately prioritize the validation experiments. We show here that, by automatically comparing and analyzing structurally similar regions of proteins of known structure interacting with a common partner, it is possible to identify mutually exclusive interactions present in the maps with a sensitivity of 70% and a specificity higher than 85% and that, in about three fourth of the correctly identified complexes, we also correctly recognize at least one residue (five on average) belonging to the interaction interface. Given the present and continuously increasing number of proteins of known structure, the requirement of the knowledge of the structure of the interacting proteins does not substantially impact on the coverage of our strategy that can be estimated to be around 25%. We also introduce here the Estrella server that embodies this strategy, is designed for users interested in validating specific hypotheses about the functional role of a protein-protein interaction and it also allows access to pre-computed data for seven organisms.

Label-Free Aptasensor for Lysozyme Detection Using Electrochemical Impedance Spectroscopy

Directory of Open Access Journals (Sweden)

Dionisia Ortiz-Aguayo

2018-01-01

Full Text Available This research develops a label-free aptamer biosensor (aptasensor based on graphite-epoxy composite electrodes (GECs for the detection of lysozyme protein using Electrochemical Impedance Spectroscopy (EIS technique. The chosen immobilization technique was based on covalent bonding using carbodiimide chemistry; for this purpose, carboxylic moieties were first generated on the graphite by electrochemical grafting. The detection was performed using [Fe(CN6]3−/[Fe(CN6]4− as redox probe. After recording the frequency response, values were fitted to its electric model using the principle of equivalent circuits. The aptasensor showed a linear response up to 5 µM for lysozyme and a limit of detection of 1.67 µM. The sensitivity of the established method was 0.090 µM−1 in relative charge transfer resistance values. The interference response by main proteins, such as bovine serum albumin and cytochrome c, has been also characterized. To finally verify the performance of the developed aptasensor, it was applied to wine analysis.

Label-Free Aptasensor for Lysozyme Detection Using Electrochemical Impedance Spectroscopy.

Science.gov (United States)

Ortiz-Aguayo, Dionisia; Del Valle, Manel

2018-01-26

This research develops a label-free aptamer biosensor (aptasensor) based on graphite-epoxy composite electrodes (GECs) for the detection of lysozyme protein using Electrochemical Impedance Spectroscopy (EIS) technique. The chosen immobilization technique was based on covalent bonding using carbodiimide chemistry; for this purpose, carboxylic moieties were first generated on the graphite by electrochemical grafting. The detection was performed using [Fe(CN)₆] 3- /[Fe(CN)₆] 4- as redox probe. After recording the frequency response, values were fitted to its electric model using the principle of equivalent circuits. The aptasensor showed a linear response up to 5 µM for lysozyme and a limit of detection of 1.67 µM. The sensitivity of the established method was 0.090 µM -1 in relative charge transfer resistance values. The interference response by main proteins, such as bovine serum albumin and cytochrome c, has been also characterized. To finally verify the performance of the developed aptasensor, it was applied to wine analysis.

An electrical bio-chip to transfer and detect electromagnetic stimulation on the cells based on vertically aligned carbon nanotubes.

Science.gov (United States)

Rafizadeh-Tafti, Saeed; Haqiqatkhah, Mohammad Hossein; Saviz, Mehrdad; Janmaleki, Mohsen; Faraji Dana, Reza; Zanganeh, Somayeh; Abdolahad, Mohammad

2017-01-01

A highly sensitive impedimetric bio-chip based on vertically aligned multiwall carbon nanotubes (VAMWCNTs), was applied in direct interaction with lung cancer cells. Our tool provided both inducing and monitoring the bioelectrical changes in the cells initiated by electromagnetic (EM) wave stimulation. EM wave of 940MHz frequency with different intensities was used. Here, wave ablation might accumulate electrical charge on the tips of nanotubes penetrated into cell's membrane. The charge might induce ionic exchanges into the cell and cause alterations in electrical states of the membrane. Transmembrane electrostatic/dynamic states would be strongly affected due to such exchanges. Our novel modality was that, the cells' vitality changes caused by charge inductions were electrically detected with the same nanotubes in the architecture of electrodes for impedance measurement. The responses of the sensor were confirmed by electron and florescent microscopy images as well as biological assays. In summation, our method provided an effective biochip for enhancing and detecting external EM stimulation on the cells useful for future diagnostic and therapeutic applications, such as wave-guided drug-resistance breakage. Copyright © 2016 Elsevier B.V. All rights reserved.

Optimizing FRET-FLIM Labeling Conditions to Detect Nuclear Protein Interactions at Native Expression Levels in Living Arabidopsis Roots

KAUST Repository

Long, Yuchen; Stahl, Yvonne; Weidtkamp-Peters, Stefanie; Smet, Wouter; Du, Yujuan; Gadella, Theodorus W. J.; Goedhart, Joachim; Scheres, Ben; Blilou, Ikram

2018-01-01

Protein complex formation has been extensively studied using Förster resonance energy transfer (FRET) measured by Fluorescence Lifetime Imaging Microscopy (FLIM). However, implementing this technology to detect protein interactions in living

78 FR 16513 - Application of Advances in Nucleic Acid and Protein Based Detection Methods to Multiplex...

Science.gov (United States)

2013-03-15

... Methods to Multiplex Detection of Transfusion- Transmissible Agents and Blood Cell Antigens in Blood... Transfusion-Transmissible Agents and Blood Cell Antigens in Blood Donations; Public Workshop AGENCY: Food and... technological advances in gene based and protein based pathogen and blood cell antigen detection methods and to...

Detection of intact megadalton protein assemblies of vanillyl-alcohol oxidase by mass spectrometry

NARCIS (Netherlands)

Berkel, van W.J.H.; Heuvel, van den R.H.H.; Versluis, C.; Heck, A.

2000-01-01

Well-resolved ion signals of intact large protein assemblies, with molecular masses extending above one million Dalton, have been detected and mass analyzed using electrospray ionization mass spectrometry, with an uncertainty in mass of <0.2&Eth;The mass spectral data seem to reflect known

Referencing cross-reactivity of detection antibodies for protein array experiments [version 1; referees: 1 approved, 2 approved with reservations

Directory of Open Access Journals (Sweden)

Darragh Lemass

2016-01-01

Full Text Available Protein arrays are frequently used to profile antibody repertoires in humans and animals. High-throughput protein array characterisation of complex antibody repertoires requires a platform-dependent, lot-to-lot validation of secondary detection antibodies. This article details the validation of an affinity-isolated anti-chicken IgY antibody produced in rabbit and a goat anti-rabbit IgG antibody conjugated with alkaline phosphatase using protein arrays consisting of 7,390 distinct human proteins. Probing protein arrays with secondary antibodies in absence of chicken serum revealed non-specific binding to 61 distinct human proteins. The cross-reactivity of the tested secondary detection antibodies points towards the necessity of platform-specific antibody characterisation studies for all secondary immunoreagents. Secondary antibody characterisation using protein arrays enables generation of reference lists of cross-reactive proteins, which can be then excluded from analysis in follow-up experiments. Furthermore, making such cross-reactivity lists accessible to the wider research community may help to interpret data generated by the same antibodies in applications not related to protein arrays such as immunoprecipitation, Western blots or other immunoassays.

CMOS image sensor for detection of interferon gamma protein interaction as a point-of-care approach.

Science.gov (United States)

Marimuthu, Mohana; Kandasamy, Karthikeyan; Ahn, Chang Geun; Sung, Gun Yong; Kim, Min-Gon; Kim, Sanghyo

2011-09-01

Complementary metal oxide semiconductor (CMOS)-based image sensors have received increased attention owing to the possibility of incorporating them into portable diagnostic devices. The present research examined the efficiency and sensitivity of a CMOS image sensor for the detection of antigen-antibody interactions involving interferon gamma protein without the aid of expensive instruments. The highest detection sensitivity of about 1 fg/ml primary antibody was achieved simply by a transmission mechanism. When photons are prevented from hitting the sensor surface, a reduction in digital output occurs in which the number of photons hitting the sensor surface is approximately proportional to the digital number. Nanoscale variation in substrate thickness after protein binding can be detected with high sensitivity by the CMOS image sensor. Therefore, this technique can be easily applied to smartphones or any clinical diagnostic devices for the detection of several biological entities, with high impact on the development of point-of-care applications.

Detection of 70 kDa heat shock protein in the saliva of dairy cows.

Science.gov (United States)

Lamy, Elsa; Jurkovich, Viktor; Rodrigues, Lénia; Geraldo, Ana; Cachucho, Liliana; Silva, Flávio; Matos, Catarina; Capela E Silva, Fernando; Pinheiro, Cristina; Könyves, László; Bakony, Mikolt; Pereira, Alfredo

2017-08-01

This Research Communication describes, for the first time, the detection of HSP70 in saliva of dairy cows. Thermal stress is a major environmental stress that limits animal growth, metabolism, and productivity. The cellular response to heat stress involves the synthesis of heat shock proteins (HSPs), presumably to protect the functional stability of cells at increasing temperatures. HSP70 has been found to be present in cattle blood serum and may also be present in other secretory fluids, such as saliva, as already observed in humans. The aim of this study was to detect heat shock protein HSP70 in bovine saliva. Saliva samples were taken from higher- (n = 5) and lower milk producing (n = 5) Holstein-Friesian cows in summer and in winter for the detection of HSP70. HSP70 concentrations were assayed using the ELISA technique. Salivary HSP70 concentrations ranged from 0·524 to 12·174 ng/ml in cows. Higher salivary HSP70 concentrations were significantly associated with higher milk production and higher environmental temperature, but not with rectal temperature.

Enhanced Missing Proteins Detection in NCI60 Cell Lines Using an Integrative Search Engine Approach.

Science.gov (United States)

Guruceaga, Elizabeth; Garin-Muga, Alba; Prieto, Gorka; Bejarano, Bartolomé; Marcilla, Miguel; Marín-Vicente, Consuelo; Perez-Riverol, Yasset; Casal, J Ignacio; Vizcaíno, Juan Antonio; Corrales, Fernando J; Segura, Victor

2017-12-01

The Human Proteome Project (HPP) aims deciphering the complete map of the human proteome. In the past few years, significant efforts of the HPP teams have been dedicated to the experimental detection of the missing proteins, which lack reliable mass spectrometry evidence of their existence. In this endeavor, an in depth analysis of shotgun experiments might represent a valuable resource to select a biological matrix in design validation experiments. In this work, we used all the proteomic experiments from the NCI60 cell lines and applied an integrative approach based on the results obtained from Comet, Mascot, OMSSA, and X!Tandem. This workflow benefits from the complementarity of these search engines to increase the proteome coverage. Five missing proteins C-HPP guidelines compliant were identified, although further validation is needed. Moreover, 165 missing proteins were detected with only one unique peptide, and their functional analysis supported their participation in cellular pathways as was also proposed in other studies. Finally, we performed a combined analysis of the gene expression levels and the proteomic identifications from the common cell lines between the NCI60 and the CCLE project to suggest alternatives for further validation of missing protein observations.

Pulse Detecting Genetic Circuit – A New Design Approach

Science.gov (United States)

Inniss, Mara; Iba, Hitoshi; Way, Jeffrey C.

2016-01-01

A robust cellular counter could enable synthetic biologists to design complex circuits with diverse behaviors. The existing synthetic-biological counters, responsive to the beginning of the pulse, are sensitive to the pulse duration. Here we present a pulse detecting circuit that responds only at the falling edge of a pulse–analogous to negative edge triggered electric circuits. As biological events do not follow precise timing, use of such a pulse detector would enable the design of robust asynchronous counters which can count the completion of events. This transcription-based pulse detecting circuit depends on the interaction of two co-expressed lambdoid phage-derived proteins: the first is unstable and inhibits the regulatory activity of the second, stable protein. At the end of the pulse the unstable inhibitor protein disappears from the cell and the second protein triggers the recording of the event completion. Using stochastic simulation we showed that the proposed design can detect the completion of the pulse irrespective to the pulse duration. In our simulation we also showed that fusing the pulse detector with a phage lambda memory element we can construct a counter which can be extended to count larger numbers. The proposed design principle is a new control mechanism for synthetic biology which can be integrated in different circuits for identifying the completion of an event. PMID:27907045

Detection of soy proteins in processed foods: Literature overview and new experimental work

NARCIS (Netherlands)

Koppelman, S.J.; Lakemond, C.M.M.; Vlooswijk, R.A.A.; Hefle, S.L.

2004-01-01

Several tests for the detection of soy proteins in foods have been described in the literature, and some are commercially available. This article gives an overview of these methods and discusses the advantages and disadvantages of each individual method. Based on the conclusions of this inventory,

Imaging the lipidome: omega-alkynyl fatty acids for detection and cellular visualization of lipid-modified proteins.

Science.gov (United States)

Hannoush, Rami N; Arenas-Ramirez, Natalia

2009-07-17

Fatty acylation or lipid modification of proteins controls their cellular activation and diverse roles in physiology. It mediates protein-protein and protein-membrane interactions and plays an important role in regulating cellular signaling pathways. Currently, there is need for visualizing lipid modifications of proteins in cells. Herein we report novel chemical probes based on omega-alkynyl fatty acids for biochemical detection and cellular imaging of lipid-modified proteins. Our study shows that omega-alkynyl fatty acids of varying chain length are metabolically incorporated onto cellular proteins. Using fluorescence imaging, we describe the subcellular distribution of lipid-modified proteins across a panel of different mammalian cell lines and during cell division. Our results demonstrate that this methodology is a useful diagnostic tool for analyzing the lipid content of cellular proteins and for studying the dynamic behavior of lipid-modified proteins in various disease or physiological states.

Detection and quantitation of twenty-seven cytokines, chemokines and growth factors pre- and post-high abundance protein depletion in human plasma

Directory of Open Access Journals (Sweden)

Seong-Beom Ahn

2014-06-01

Full Text Available Cytokines, chemokines and growth factors (CCGFs in human plasma are analyzed for identification of biomarkers. However concentrations of CCGFs are very low; it is difficult to identify and quantify low abundance proteins in the presence of the high abundance proteins (HAPs unless HAPs are removed prior to analysis. However, there is a concern that the low abundance proteins such as CCGFs may also be removed during the HAP depletion process. In this study, we have examined whether or not depletion of the HAPs enhances detection of the CCGFs by immuno-assays. Top 14 HAPs were depleted from 10 healthy volunteers’ plasma using MARS-14 immuno-depletion column and a total of 27 CCGFs were analyzed by bead-based multiplexed immuno-assay. All 27 CCGFs were detected in neat plasma (NP, 25 were detected in flow through fraction (FT and 21 were detected in bound protein (BP fraction. Concentrations of 22 CCGFs were significantly higher in NP compared to FT and BP. Only one CCGF had higher concentration in FT compared to NP. The remaining 2 CCGFs were not different between NP and FT. It was counter-productive for the detection of 24 CCGFs after HAP removal, primarily due to post-depletion protein precipitation and/or re-suspension of pellets.

An Amperometric Immunosensor Based on Graphene Composite Film and Protein a for Chlorpyrifos Detection

Directory of Open Access Journals (Sweden)

Xiangyou Wang

2014-09-01

Full Text Available In this paper, an immunosensor was designed for chlorpyrifos detection, which was based on graphene-multi-walled carbon nanotubes-gold nanoparticle-chitosan (GR-MWCNTs-AuNPs- CHIT nanocomposite film. Protein A (SPA can combine with gold nanoparticles, which made anti- chlorpyrifos antibody immobilized orientedly, eventually the modified immunosensor was developed for the detection of chlorpyrifos residues. Under the optimized conditions, a regression equation: y=9.5676 lgC (ng/mL +18.164 (R2=0.9976 was obtained with a detection limit as low as 0.037 ng/mL. The proposed chlorpyrifos immunosensor exhibited high reproducibility, stability, and good selectivity and regeneration, it has the potential of real sample detection.

Electrical spin injection and detection in silicon nanowires with axial doping gradient.

Science.gov (United States)

Kountouriotis, Konstantinos; Barreda, Jorge L; Keiper, Timothy David; Zhang, Mei; Xiong, Peng

2018-06-13

The interest in spin transport in nanoscopic semiconductor channels is driven by both the inevitable miniaturization of spintronics devices toward nanoscale and the rich spin-dependent physics the quantum confinement engenders. For such studies, the all-important issue of the ferromagnet/semiconductor (FM/SC) interface becomes even more critical at nanoscale. Here we elucidate the effects of the FM/SC interface on electrical spin injection and detection at nanoscale dimensions, utilizing a unique type of Si nanowires (NWs) with an inherent axial doping gradient. Two-terminal and nonlocal four-terminal lateral spin-valve measurements were performed using different combinations from a series of FM contacts positioned along the same NW. The data are analyzed with a general model of spin accumulation in a normal channel under electrical spin injection from a FM, which reveals a distinct correlation of decreasing spin-valve signal with increasing injector junction resistance. The observation is attributed to the diminishing contribution of the d-electrons in the FM to the injected current spin polarization with increasing Schottky barrier width. The results demonstrate that there is a window of interface parameters for optimal spin injection efficiency and current spin polarization, which provides important design guidelines for nano-spintronic devices with quasi-1D semiconductor channels.

Dual-gate polysilicon nanoribbon biosensors enable high sensitivity detection of proteins

International Nuclear Information System (INIS)

Zeimpekis, I; Sun, K; Hu, C; Ditshego, N M J; De Planque, M R R; Chong, H M H; Morgan, H; Ashburn, P; Thomas, O

2016-01-01

We demonstrate the advantages of dual-gate polysilicon nanoribbon biosensors with a comprehensive evaluation of different measurement schemes for pH and protein sensing. In particular, we compare the detection of voltage and current changes when top- and bottom-gate bias is applied. Measurements of pH show that a large voltage shift of 491 mV pH"−"1 is obtained in the subthreshold region when the top-gate is kept at a fixed potential and the bottom-gate is varied (voltage sweep). This is an improvement of 16 times over the 30 mV pH"−"1 measured using a top-gate sweep with the bottom-gate at a fixed potential. A similar large voltage shift of 175 mV is obtained when the protein avidin is sensed using a bottom-gate sweep. This is an improvement of 20 times compared with the 8.8 mV achieved from a top-gate sweep. Current measurements using bottom-gate sweeps do not deliver the same signal amplification as when using bottom-gate sweeps to measure voltage shifts. Thus, for detecting a small signal change on protein binding, it is advantageous to employ a double-gate transistor and to measure a voltage shift using a bottom-gate sweep. For top-gate sweeps, the use of a dual-gate transistor enables the current sensitivity to be enhanced by applying a negative bias to the bottom-gate to reduce the carrier concentration in the nanoribbon. For pH measurements, the current sensitivity increases from 65% to 149% and for avidin sensing it increases from 1.4% to 2.5%. (paper)

Virus detection and quantification using electrical parameters

Science.gov (United States)

Ahmad, Mahmoud Al; Mustafa, Farah; Ali, Lizna M.; Rizvi, Tahir A.

2014-10-01

Here we identify and quantitate two similar viruses, human and feline immunodeficiency viruses (HIV and FIV), suspended in a liquid medium without labeling, using a semiconductor technique. The virus count was estimated by calculating the impurities inside a defined volume by observing the change in electrical parameters. Empirically, the virus count was similar to the absolute value of the ratio of the change of the virus suspension dopant concentration relative to the mock dopant over the change in virus suspension Debye volume relative to mock Debye volume. The virus type was identified by constructing a concentration-mobility relationship which is unique for each kind of virus, allowing for a fast (within minutes) and label-free virus quantification and identification. For validation, the HIV and FIV virus preparations were further quantified by a biochemical technique and the results obtained by both approaches corroborated well. We further demonstrate that the electrical technique could be applied to accurately measure and characterize silica nanoparticles that resemble the virus particles in size. Based on these results, we anticipate our present approach to be a starting point towards establishing the foundation for label-free electrical-based identification and quantification of an unlimited number of viruses and other nano-sized particles.

Attomolar detection of proteins via cascade strand-displacement amplification and polystyrene nanoparticle enhancement in fluorescence polarization aptasensors.

Science.gov (United States)

Huang, Yong; Liu, Xiaoqian; Huang, Huakui; Qin, Jian; Zhang, Liangliang; Zhao, Shulin; Chen, Zhen-Feng; Liang, Hong

2015-08-18

Extremely sensitive and accurate measurements of protein markers for early detection and monitoring of diseases pose a formidable challenge. Herein, we develop a new type of amplified fluorescence polarization (FP) aptasensor based on allostery-triggered cascade strand-displacement amplification (CSDA) and polystyrene nanoparticle (PS NP) enhancement for ultrasensitive detection of proteins. The assay system consists of a fluorescent dye-labeled aptamer hairpin probe and a PS NP-modified DNA duplex (assistant DNA/trigger DNA duplex) probe with a single-stranded part and DNA polymerase. Two probes coexist stably in the absence of target, and the dye exhibits relatively low FP background. Upon recognition and binding with a target protein, the stem of the aptamer hairpin probe is opened, after which the opened hairpin probe hybridizes with the single-stranded part in the PS NP-modified DNA duplex probe and triggers the CSDA reaction through the polymerase-catalyzed recycling of both target protein and trigger DNA. Throughout this CSDA process, numerous massive dyes are assembled onto PS NPs, which results in a substantial FP increase that provides a readout signal for the amplified sensing process. Our newly proposed amplified FP aptasensor enables the quantitative measurement of proteins with the detection limit in attomolar range, which is about 6 orders of magnitude lower than that of traditional homogeneous aptasensors. Moreover, this sensing method also exhibits high specificity for target proteins and can be performed in homogeneous solutions. In addition, the suitability of this method for the quantification of target protein in biological samples has also been shown. Considering these distinct advantages, the proposed sensing method can be expected to provide an ultrasensitive platform for the analysis of various types of target molecules.

Electricity sequence control

International Nuclear Information System (INIS)

Shin, Heung Ryeol

2010-03-01

The contents of the book are introduction of control system, like classification and control signal, introduction of electricity power switch, such as push-button and detection switch sensor for induction type and capacitance type machinery for control, solenoid valve, expression of sequence and type of electricity circuit about using diagram, time chart, marking and term, logic circuit like Yes, No, and, or and equivalence logic, basic electricity circuit, electricity sequence control, added condition, special program control about choice and jump of program, motor control, extra circuit on repeat circuit, pause circuit in a conveyer, safety regulations and rule about classification of electricity disaster and protective device for insulation.

Cervical cancer detection by electrical impedance in a Colombian setting

International Nuclear Information System (INIS)

A Miranda, David; P Corzo, Sandra; Correa, C A González

2013-01-01

Electrical properties of normal and neoplastic cervical tissues in a heterogeneous group of 56 Colombian women were studied by electrical impedance spectroscopy and a model based on the Generalized Effective-Medium Theory of Induced Polarization (GEMTIP). Differences between the electrical bioimpedance spectra were correlated with cellular and tissue parameters. The analysis performed by the proposed model suggest that the number of different types of cellular layers that form the biological tissue, the intracellular and extracellular conductivity could be used to explain the differences between electrical bioimpedance spectra in normal and neoplastic tissues.

Prediction of Protein-Protein Interactions Related to Protein Complexes Based on Protein Interaction Networks

Directory of Open Access Journals (Sweden)

Peng Liu

2015-01-01

Full Text Available A method for predicting protein-protein interactions based on detected protein complexes is proposed to repair deficient interactions derived from high-throughput biological experiments. Protein complexes are pruned and decomposed into small parts based on the adaptive k-cores method to predict protein-protein interactions associated with the complexes. The proposed method is adaptive to protein complexes with different structure, number, and size of nodes in a protein-protein interaction network. Based on different complex sets detected by various algorithms, we can obtain different prediction sets of protein-protein interactions. The reliability of the predicted interaction sets is proved by using estimations with statistical tests and direct confirmation of the biological data. In comparison with the approaches which predict the interactions based on the cliques, the overlap of the predictions is small. Similarly, the overlaps among the predicted sets of interactions derived from various complex sets are also small. Thus, every predicted set of interactions may complement and improve the quality of the original network data. Meanwhile, the predictions from the proposed method replenish protein-protein interactions associated with protein complexes using only the network topology.

Polyethylenimine-coated Fe3O4 nanoparticles effectively quench fluorescent DNA, which can be developed as a novel platform for protein detection.

Science.gov (United States)

Ma, Long; Sun, Nana; Zhang, Jinyan; Tu, Chunhao; Cao, Xiuqi; Duan, Demin; Diao, Aipo; Man, Shuli

2017-11-23

We report a novel assembly of polyethyleneimine (PEI)-coated Fe 3 O 4 nanoparticles (NPs) with single-stranded DNA (ssDNA), and the fluorescence of the dye labeled in the DNA is remarkably quenched. In the presence of a target protein, the protein-DNA aptamer mutual interaction releases the ssDNA from this assembly and hence restores the fluorescence. This feature could be adopted to develop an aptasensor for protein detection. As a proof-of-concept, for the first time, we have used this proposed sensing strategy to detect thrombin selectively and sensitively. Furthermore, simultaneous multiple detection of thrombin and lysozyme in a complex protein mixture has been proven to be possible.

Nitrogen-detected TROSY yields comparable sensitivity to proton-detected TROSY for non-deuterated, large proteins under physiological salt conditions

Energy Technology Data Exchange (ETDEWEB)

Takeuchi, Koh [National Institute for Advanced Industrial Science and Technology, Molecular Profiling Research Center for Drug Discovery (Japan); Arthanari, Haribabu [Harvard Medical School, Department of Biochemistry and Molecular Pharmacology (United States); Imai, Misaki [Japan Biological Informatics Consortium, Research and Development Department (Japan); Wagner, Gerhard, E-mail: gerhard-wagner@hms.harvard.edu [Harvard Medical School, Department of Biochemistry and Molecular Pharmacology (United States); Shimada, Ichio, E-mail: shimada@iw-nmr.f.u-tokyo.ac.jp [National Institute for Advanced Industrial Science and Technology, Molecular Profiling Research Center for Drug Discovery (Japan)

2016-02-15

Direct detection of the TROSY component of proton-attached {sup 15}N nuclei ({sup 15}N-detected TROSY) yields high quality spectra with high field magnets, by taking advantage of the slow {sup 15}N transverse relaxation. The slow transverse relaxation and narrow line width of the {sup 15}N-detected TROSY resonances are expected to compensate for the inherently low {sup 15}N sensitivity. However, the sensitivity of {sup 15}N-detected TROSY in a previous report was one-order of magnitude lower than in the conventional {sup 1}H-detected version. This could be due to the fact that the previous experiments were performed at low salt (0–50 mM), which is advantageous for {sup 1}H-detected experiments. Here, we show that the sensitivity gap between {sup 15}N and {sup 1}H becomes marginal for a non-deuterated, large protein (τ{sub c} = 35 ns) at a physiological salt concentration (200 mM). This effect is due to the high salt tolerance of the {sup 15}N-detected TROSY. Together with the previously reported benefits of the {sup 15}N-detected TROSY, our results provide further support for the significance of this experiment for structural studies of macromolecules when using high field magnets near and above 1 GHz.

Protein biomarkers on tissue as imaged via MALDI mass spectrometry: A systematic approach to study the limits of detection.

Science.gov (United States)

van de Ven, Stephanie M W Y; Bemis, Kyle D; Lau, Kenneth; Adusumilli, Ravali; Kota, Uma; Stolowitz, Mark; Vitek, Olga; Mallick, Parag; Gambhir, Sanjiv S

2016-06-01

MALDI mass spectrometry imaging (MSI) is emerging as a tool for protein and peptide imaging across tissue sections. Despite extensive study, there does not yet exist a baseline study evaluating the potential capabilities for this technique to detect diverse proteins in tissue sections. In this study, we developed a systematic approach for characterizing MALDI-MSI workflows in terms of limits of detection, coefficients of variation, spatial resolution, and the identification of endogenous tissue proteins. Our goal was to quantify these figures of merit for a number of different proteins and peptides, in order to gain more insight in the feasibility of protein biomarker discovery efforts using this technique. Control proteins and peptides were deposited in serial dilutions on thinly sectioned mouse xenograft tissue. Using our experimental setup, coefficients of variation were biomarkers and a new benchmarking strategy that can be used for comparing diverse MALDI-MSI workflows. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Protein Self-Assemblies That Can Generate, Hold, and Discharge Electric Potential in Response to Changes in Relative Humidity.

Science.gov (United States)

Carter, Nathan A; Grove, Tijana Z

2018-05-30

Generation of electric potential upon external stimulus has attracted much attention for the development of highly functional sensors and devices. Herein, we report large-displacement, fast actuation in the self-assembled engineered repeat protein Consensus Tetratricopeptide Repeat protein (CTPR18) materials. The ionic nature of the CTPR18 protein coupled to the long-range alignment upon self-assembly results in the measured conductivity of 7.1 × 10 -2 S cm -1 , one of the highest reported for protein materials. The change of through-thickness morphological gradient in the self-assembled materials provides the means to select between faster, highly water-sensitive actuation or vastly increased mechanical strength. Tuning of the mode of motion, e.g., bending, twisting, and folding, is achieved by changing the morphological director. We further show that the highly ionic character of CTPR18 gives rise to piezo-like behavior in these materials, exemplified by low-voltage, ionically driven actuation and mechanically driven generation/discharge of voltage. This work contributes to our understanding of the emergence of stimuli-responsiveness in biopolymer assemblies.

Sensitive detection of C-reactive protein using optical fiber Bragg gratings.

Science.gov (United States)

Sridevi, S; Vasu, K S; Asokan, S; Sood, A K

2015-03-15

An accurate and highly sensitive sensor platform has been demonstrated for the detection of C-reactive protein (CRP) using optical fiber Bragg gratings (FBGs). The CRP detection has been carried out by monitoring the shift in Bragg wavelength (ΔλB) of an etched FBG (eFBG) coated with an anti-CRP antibody (aCRP)-graphene oxide (GO) complex. The complex is characterized by Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy and atomic force microscopy. A limit of detection of 0.01mg/L has been achieved with a linear range of detection from 0.01mg/L to 100mg/L which includes clinical range of CRP. The eFBG sensor coated with only aCRP (without GO) show much less sensitivity than that of aCRP-GO complex coated eFBG. The eFBG sensors show high specificity to CRP even in the presence of other interfering factors such as urea, creatinine and glucose. The affinity constant of ∼1.1×10(10)M(-1) has been extracted from the data of normalized shift (ΔλB/λB) as a function of CRP concentration. Copyright © 2014 Elsevier B.V. All rights reserved.

Nano-mole scale sequential signal assignment by 1 H-detected protein solid-state NMR

KAUST Repository

Wang, Songlin; Parthasarathy, Sudhakar; Xiao, Yiling; Nishiyama, Yusuke; Long, Fei; Matsuda, Isamu; Endo, Yuki; Nemoto, Takahiro; Yamauchi, Kazuo; Asakura, Tetsuo; Takeda, Mitsuhiro; Terauchi, Tsutomu; Kainosho, Masatsune; Ishii, Yoshitaka

2015-01-01

We present a 3D 1H-detected solid-state NMR (SSNMR) approach for main-chain signal assignments of 10-100 nmol of fully protonated proteins using ultra-fast magic-angle spinning (MAS) at ∼80 kHz by a novel spectral-editing method, which permits drastic spectral simplification. The approach offers ∼110 fold time saving over a traditional 3D 13C-detected SSNMR approach. This journal is © The Royal Society of Chemistry 2015.

Protein Detection Using the Multiplexed Proximity Extension Assay (PEA) from Plasma and Vaginal Fluid Applied to the Indicating FTA Elute Micro Card™

Science.gov (United States)

Berggrund, Malin; Ekman, Daniel; Gustavsson, Inger; Sundfeldt, Karin; Olovsson, Matts; Enroth, Stefan; Gyllensten, Ulf

2016-01-01

The indicating FTA elute micro card™ has been developed to collect and stabilize the nucleic acid in biological samples and is widely used in human and veterinary medicine and other disciplines. This card is not recommended for protein analyses, since surface treatment may denature proteins. We studied the ability to analyse proteins in human plasma and vaginal fluid as applied to the indicating FTA elute micro card™ using the sensitive proximity extension assay (PEA). Among 92 proteins in the Proseek Multiplex Oncology Iv2 panel, 87 were above the limit of detection (LOD) in liquid plasma and 56 among 92 above LOD in plasma applied to FTA cards. Washing and protein elution protocols were compared to identify an optimal method. Liquid-based cytology samples showed a lower number of proteins above LOD than FTA cards with vaginal fluid samples applied. Our results demonstrate that samples applied to the indicating FTA elute micro card™ are amendable to protein analyses, given that a sensitive protein detection assay is used. The results imply that biological samples applied to FTA cards can be used for DNA, RNA and protein detection. PMID:28936257

Protein Detection Using the Multiplexed Proximity Extension Assay (PEA from Plasma and Vaginal Fluid Applied to the Indicating FTA Elute Micro Card™

Directory of Open Access Journals (Sweden)

Malin Berggrund

2016-01-01

Full Text Available The indicating FTA elute micro card™ has been developed to collect and stabilize the nucleic acid in biological samples and is widely used in human and veterinary medicine and other disciplines. This card is not recommended for protein analyses, since surface treatment may denature proteins. We studied the ability to analyse proteins in human plasma and vaginal fluid as applied to the indicating FTA elute micro card™ using the sensitive proximity extension assay (PEA. Among 92 proteins in the Proseek Multiplex Oncology Iv2 panel, 87 were above the limit of detection (LOD in liquid plasma and 56 among 92 above LOD in plasma applied to FTA cards. Washing and protein elution protocols were compared to identify an optimal method. Liquid-based cytology samples showed a lower number of proteins above LOD than FTA cards with vaginal fluid samples applied. Our results demonstrate that samples applied to the indicating FTA elute micro card™ are amendable to protein analyses, given that a sensitive protein detection assay is used. The results imply that biological samples applied to FTA cards can be used for DNA, RNA and protein detection.

Protein Detection Using the Multiplexed Proximity Extension Assay (PEA) from Plasma and Vaginal Fluid Applied to the Indicating FTA Elute Micro Card™.

Science.gov (United States)

Berggrund, Malin; Ekman, Daniel; Gustavsson, Inger; Sundfeldt, Karin; Olovsson, Matts; Enroth, Stefan; Gyllensten, Ulf

2016-01-01

The indicating FTA elute micro card™ has been developed to collect and stabilize the nucleic acid in biological samples and is widely used in human and veterinary medicine and other disciplines. This card is not recommended for protein analyses, since surface treatment may denature proteins. We studied the ability to analyse proteins in human plasma and vaginal fluid as applied to the indicating FTA elute micro card™ using the sensitive proximity extension assay (PEA). Among 92 proteins in the Proseek Multiplex Oncology Iv2 panel, 87 were above the limit of detection (LOD) in liquid plasma and 56 among 92 above LOD in plasma applied to FTA cards. Washing and protein elution protocols were compared to identify an optimal method. Liquid-based cytology samples showed a lower number of proteins above LOD than FTA cards with vaginal fluid samples applied. Our results demonstrate that samples applied to the indicating FTA elute micro card™ are amendable to protein analyses, given that a sensitive protein detection assay is used. The results imply that biological samples applied to FTA cards can be used for DNA, RNA and protein detection.

Slip Ratio Estimation and Regenerative Brake Control for Decelerating Electric Vehicles without Detection of Vehicle Velocity and Acceleration

Science.gov (United States)

Suzuki, Toru; Fujimoto, Hiroshi

In slip ratio control systems, it is necessary to detect the vehicle velocity in order to obtain the slip ratio. However, it is very difficult to measure this velocity directly. We have proposed slip ratio estimation and control methods that do not require the vehicle velocity with acceleration. In this paper, the slip ratio estimation and control methods are proposed without detecting the vehicle velocity and acceleration when it is decelerating. We carried out simulations and experiments by using an electric vehicle to verify the effectiveness of the proposed method.

Electrical resistivity imaging survey to detect uncharted mine galleries in the mining district of Linares, Jaén, Spain

Science.gov (United States)

Martínez-López, J.; Rey, J.; Dueñas, J.; Hidalgo, C.; Benavente, J.

2012-02-01

The scarcity of information about the existence of old mining shafts and galleries in urban areas is an important issue for future urban development. Electrical resistivity tomography is a non-destructive geophysical technique that can detect and characterize such subsurface cavities based on differences in the behaviour of electrical current in the void and in the embedding rock. Here we present a study in which this technique was used to determine the location of old engineered structures around the city of Linares, southern Spain, and to relate these structures to the abandoned deep mines present in the area. Eight electrical resistivity imaging profiles were performed, with a total of 22 808 measurements. Correlations between geoelectrical anomalies allow detection of the depth and the direction of several galleries, as well as the voids that result from mining extraction. Given the depth at which these structures are located (in some cases less than 5 m), they pose an important risk for future construction projects in areas of urban expansion. This technique is shown to be a useful tool for locating areas that pose important urban risks and, by extension, for the decision-making process in territorial planning, especially in areas with a history of deep mining.

Electrical resistivity imaging survey to detect uncharted mine galleries in the mining district of Linares, Jaén, Spain

International Nuclear Information System (INIS)

Martínez-López, J; Rey, J; Hidalgo, C; Dueñas, J; Benavente, J

2012-01-01

The scarcity of information about the existence of old mining shafts and galleries in urban areas is an important issue for future urban development. Electrical resistivity tomography is a non-destructive geophysical technique that can detect and characterize such subsurface cavities based on differences in the behaviour of electrical current in the void and in the embedding rock. Here we present a study in which this technique was used to determine the location of old engineered structures around the city of Linares, southern Spain, and to relate these structures to the abandoned deep mines present in the area. Eight electrical resistivity imaging profiles were performed, with a total of 22 808 measurements. Correlations between geoelectrical anomalies allow detection of the depth and the direction of several galleries, as well as the voids that result from mining extraction. Given the depth at which these structures are located (in some cases less than 5 m), they pose an important risk for future construction projects in areas of urban expansion. This technique is shown to be a useful tool for locating areas that pose important urban risks and, by extension, for the decision-making process in territorial planning, especially in areas with a history of deep mining

Geophysical techniques in detection to river embankments - A case study: To locate sites of potential leaks using surface-wave and electrical methods

Science.gov (United States)

Chen, C.; Liu, J.; Xu, S.; Xia, J.; ,

2004-01-01

Geophysical technologies are very effective in environmental, engineering and groundwater applications. Parameters of delineating nature of near-surface materials such as compressional-wave velocity, shear-wave velocity can be obtained using shallow seismic methods. Electric methods are primary approaches for investigating groundwater and detecting leakage. Both of methods are applied to detect embankment in hope of obtaining evidences of the strength and moisture inside the body. A technological experiment has done for detecting and discovering the hidden troubles in the embankment of Yangtze River, Songzi, Hubei, China in 2003. Surface-wave and DC multi-channel array resistivity sounding techniques were used to detect hidden trouble inside and under dike like pipe-seeps. This paper discusses the exploration strategy and the effect of geological characteristics. A practical approach of combining seismic and electric resistivity measurements was applied to locate potential pipe-seeps in embankment in the experiment. The method presents a potential leak factor based on the shear-wave velocity and the resistivity of the medium to evaluate anomalies. An anomaly found in a segment of embankment detected was verified, where occurred a pipe-seep during the 98' flooding.

Study of hydrogenated amorphous silicon devices under intense electric field: application to nuclear detection; Etude de dispositifs electroniques en silicium amorphe hydrogene sous fort champ electrique: application a la detection nucleaire

Energy Technology Data Exchange (ETDEWEB)

Ilie, A. [CEA Centre d`Etudes de Saclay, 91 - Gif-sur-Yvette (France). Direction des Technologies Avancees]|[Paris-11 Univ., 91 - Orsay (France)

1996-12-31

The goal of this work was the study, development and optimization of hydrogenated amorphous silicon (a-Si:H) devices for use in detection of ionizing radiation. Thick p-i-n devices, capable of withstanding large electric fields (up to 10{sup 6} V/cm) with small currents (nA/cm{sup 2}), were developed. To decrease fabrication time, films were made using the `He diluted` PECVD process and compared to standard a-Si:H films. Aspects connected to specific detector applications as well as to the fundamental physics of a-Si:H were considered: the internal electric field technique, in which the depletion charge was measured as a function of the applied bias voltage; study of the leakage current of p-i-n devices permitted us to demonstrate different regimes: depletion, field-enhanced thermal generation and electronic injection across the p layer. The effect of the electric field on the thermal generation of the carriers was studied considering the Poole-Frenkel and tunneling mechanisms. A model was developed taking under consideration the statistics of the correlated states and electron-phonon coupling. The results suggest that mechanisms not included in the `standard model` of a Si:h need to be considered, such as defect relaxation, a filed-dependent mobility edge etc...; a new metastable phenomenon, induced by prolonged exposure to a strong electric field, was observed and studied. It is characterized by marked decrease of the leakage current and the detector noise, and increase in the breakdown voltage, as well as an improvement of carrier collection efficiency. This forming process appears to be principally due to an activation of the dopants in the p layer; finally, the capacity of thick p-i-n a Si:H devices to detect ionizing radiation has been evaluated. We show that it is possible, with 20-50 micron thick p-i-n devices, to detect the full spectrum of alpha and beta particles. With an appropriate converter, neutron detection then becomes possible. (author). 137 refs.

Detecting electroporation by assessing the time constants in the exponential response of human skin to voltage controlled impulse electrical stimulation.

Science.gov (United States)

Bîrlea, Sinziana I; Corley, Gavin J; Bîrlea, Nicolae M; Breen, Paul P; Quondamatteo, Fabio; OLaighin, Gearóid

2009-01-01

We propose a new method for extracting the electrical properties of human skin based on the time constant analysis of its exponential response to impulse stimulation. As a result of this analysis an adjacent finding has arisen. We have found that stratum corneum electroporation can be detected using this analysis method. We have observed that a one time-constant model is appropriate for describing the electrical properties of human skin at low amplitude applied voltages (30V). Higher voltage amplitudes (>30V) have been proven to create pores in the skin's stratum corneum which offer a new, lower resistance, pathway for the passage of current through the skin. Our data shows that when pores are formed in the stratum corneum they can be detected, in-vivo, due to the fact that a second time constant describes current flow through them.

Detection of Nuclear Protein Profile Changes by Human Metapneumovirus M2-2 Protein Using Quantitative Differential Proteomics

Directory of Open Access Journals (Sweden)

Yuping Ren

2017-12-01

Full Text Available Human metapneumovirus (hMPV is a leading cause of lower respiratory infection in pediatric populations globally. This study examined proteomic profile changes in A549 cells infected with hMPV and two attenuated mutants with deleted PDZ domain-binding motif(s in the M2-2 protein. These motifs are involved in the interruption of antiviral signaling, namely the interaction between the TNF receptor associated factor (TRAF and mitochondrial antiviral-signaling (MAVS proteins. The aim of this study was to provide insight into the overall and novel impact of M2-2 motifs on cellular responses via an unbiased comparison. Tandem mass tagging, stable isotope labeling, and high-resolution mass spectrometry were used for quantitative proteomic analysis. Using quantitative proteomics and Venn analysis, 1248 common proteins were detected in all infected samples of both technical sets. Hierarchical clustering of the differentiated proteome displayed distinct proteomic signatures that were controlled by the motif(s. Bioinformatics and experimental analysis confirmed the differentiated proteomes, revealed novel cellular biological events, and implicated key pathways controlled by hMPV M2-2 PDZ domain-binding motif(s. This provides further insight for evaluating M2-2 mutants as potent vaccine candidates.

Detection of phosphorylated mitogen-activated protein kinase in the developing spinal cord of the mouse embryo

International Nuclear Information System (INIS)

Teraishi, Toshiya; Miura, Kenji

2011-01-01

Highlights: → We detected physiologically phosphorylated MAPKs in developing spinal cord. → We detected physiologically phosphorylated MAPKs by an improved method. → p-ERK1/2 and p-JNK1/2 were detected in the marginal layer and the dorsal horn. → p-ERK1/2 and p-JNK1/2 might play critical roles in the developing spinal cord. → Constructing phosphoprotein atlases will be possible if expanding this work. -- Abstract: Global understanding of the proteome is a major research topic. The comprehensive visualization of the distribution of proteins in vivo or the construction of in situ protein atlases may be a valuable strategy for proteomic researchers. Information about the distribution of various proteins under physiological and pathological conditions should be extremely valuable for the basic and clinical sciences. The mitogen-activated protein kinase (MAPK) cascade plays an essential role in intracellular signaling in organisms. This cascade also regulates biological processes involving development, differentiation, and proliferation. Phosphorylation and dephosphorylation are integral reactions in regulating the activity of MAPKs. Changes in the phosphorylation state of MAPKs are rapid and reversible; therefore, the localizations of physiologically phosphorylated MAPKs in vivo are difficult to accurately detect. Furthermore, phosphorylated MAPKs are likely to change phosphorylated states through commonly used experimental manipulations. In the present study, as a step toward the construction of in situ phosphoprotein atlases, we attempted to detect physiologically phosphorylated MAPKs in vivo in developing spinal cords of mice. We previously reported an improved immunohistochemical method for detecting unstable phosphorylated MAPKs. The distribution patterns of phosphorylated MAPKs in the spinal cords of embryonic mice from embryonic day 13 (E13) to E17 were observed with an improved immunohistochemical method. Phosphorylated extracellular signal

Change detection in the dynamics of an intracellular protein synthesis model using nonlinear Kalman filtering.

Science.gov (United States)

Rigatos, Gerasimos G; Rigatou, Efthymia G; Djida, Jean Daniel

2015-10-01

A method for early diagnosis of parametric changes in intracellular protein synthesis models (e.g. the p53 protein - mdm2 inhibitor model) is developed with the use of a nonlinear Kalman Filtering approach (Derivative-free nonlinear Kalman Filter) and of statistical change detection methods. The intracellular protein synthesis dynamic model is described by a set of coupled nonlinear differential equations. It is shown that such a dynamical system satisfies differential flatness properties and this allows to transform it, through a change of variables (diffeomorphism), to the so-called linear canonical form. For the linearized equivalent of the dynamical system, state estimation can be performed using the Kalman Filter recursion. Moreover, by applying an inverse transformation based on the previous diffeomorphism it becomes also possible to obtain estimates of the state variables of the initial nonlinear model. By comparing the output of the Kalman Filter (which is assumed to correspond to the undistorted dynamical model) with measurements obtained from the monitored protein synthesis system, a sequence of differences (residuals) is obtained. The statistical processing of the residuals with the use of x2 change detection tests, can provide indication within specific confidence intervals about parametric changes in the considered biological system and consequently indications about the appearance of specific diseases (e.g. malignancies).

Detecting rapid mass movements using electrical self-potential measurements

Science.gov (United States)

Heinze, Thomas; Limbrock, Jonas; Pudasaini, Shiva P.; Kemna, Andreas

2017-04-01

Rapid mass movements are a latent danger for lives and infrastructure in almost any part of the world. Often such mass movements are caused by increasing pore pressure, for example, landslides after heavy rainfall or dam breaking after intrusion of water in the dam. Among several other geophysical methods used to observe water movement, the electrical self-potential method has been applied to a broad range of monitoring studies, especially focusing on volcanism and dam leakage but also during hydraulic fracturing and for earthquake prediction. Electrical self-potential signals may be caused by various mechanisms. Though, the most relevant source of the self-potential field in the given context is the streaming potential, caused by a flowing electrolyte through porous media with electrically charged internal surfaces. So far, existing models focus on monitoring water flow in non-deformable porous media. However, as the self-potential is sensitive to hydraulic parameters of the soil, any change in these parameters will cause an alteration of the electric signal. Mass movement will significantly influence the hydraulic parameters of the solid as well as the pressure field, assuming that fluid movement is faster than the pressure diffusion. We will present results of laboratory experiments under drained and undrained conditions with fluid triggered as well as manually triggered mass movements, monitored with self-potential measurements. For the undrained scenarios, we observe a clear correlation between the mass movements and signals in the electric potential, which clearly differ from the underlying potential variations due to increased saturation and fluid flow. In the drained experiments, we do not observe any measurable change in the electric potential. We therefore assume that change in fluid properties and release of the load causes disturbances in flow and streaming potential. We will discuss results of numerical simulations reproducing the observed effect. Our

Electrical detection of spin transport in Si two-dimensional electron gas systems

Science.gov (United States)

Chang, Li-Te; Fischer, Inga Anita; Tang, Jianshi; Wang, Chiu-Yen; Yu, Guoqiang; Fan, Yabin; Murata, Koichi; Nie, Tianxiao; Oehme, Michael; Schulze, Jörg; Wang, Kang L.

2016-09-01

Spin transport in a semiconductor-based two-dimensional electron gas (2DEG) system has been attractive in spintronics for more than ten years. The inherent advantages of high-mobility channel and enhanced spin-orbital interaction promise a long spin diffusion length and efficient spin manipulation, which are essential for the application of spintronics devices. However, the difficulty of making high-quality ferromagnetic (FM) contacts to the buried 2DEG channel in the heterostructure systems limits the potential developments in functional devices. In this paper, we experimentally demonstrate electrical detection of spin transport in a high-mobility 2DEG system using FM Mn-germanosilicide (Mn(Si0.7Ge0.3)x) end contacts, which is the first report of spin injection and detection in a 2DEG confined in a Si/SiGe modulation doped quantum well structure (MODQW). The extracted spin diffusion length and lifetime are l sf = 4.5 μm and {τ }{{s}}=16 {{ns}} at 1.9 K respectively. Our results provide a promising approach for spin injection into 2DEG system in the Si-based MODQW, which may lead to innovative spintronic applications such as spin-based transistor, logic, and memory devices.

Non-local electrical spin injection and detection in germanium at room temperature

Science.gov (United States)

Rortais, F.; Vergnaud, C.; Marty, A.; Vila, L.; Attané, J.-P.; Widiez, J.; Zucchetti, C.; Bottegoni, F.; Jaffrès, H.; George, J.-M.; Jamet, M.

2017-10-01

Non-local carrier injection/detection schemes lie at the very foundation of information manipulation in integrated systems. This paradigm consists in controlling with an external signal the channel where charge carriers flow between a "source" and a well separated "drain." The next generation electronics may operate on the spin of carriers in addition to their charge and germanium appears as the best hosting material to develop such a platform for its compatibility with mainstream silicon technology and the predicted long electron spin lifetime at room temperature. In this letter, we demonstrate injection of pure spin currents (i.e., with no associated transport of electric charges) in germanium, combined with non-local spin detection at 10 K and room temperature. For this purpose, we used a lateral spin valve with epitaxially grown magnetic tunnel junctions as spin injector and spin detector. The non-local magnetoresistance signal is clearly visible and reaches ≈15 mΩ at room temperature. The electron spin lifetime and diffusion length are 500 ps and 1 μm, respectively, the spin injection efficiency being as high as 27%. This result paves the way for the realization of full germanium spintronic devices at room temperature.

Combination of atomic force microscopy and mass spectrometry for the detection of target protein in the serum samples of children with autism spectrum disorders

Science.gov (United States)

Kaysheva, A. L.; Pleshakova, T. O.; Kopylov, A. T.; Shumov, I. D.; Iourov, I. Y.; Vorsanova, S. G.; Yurov, Y. B.; Ziborov, V. S.; Archakov, A. I.; Ivanov, Y. D.

2017-10-01

Possibility of detection of target proteins associated with development of autistic disorders in children with use of combined atomic force microscopy and mass spectrometry (AFM/MS) method is demonstrated. The proposed method is based on the combination of affine enrichment of proteins from biological samples and visualization of these proteins by AFM and MS analysis with quantitative detection of target proteins.

Desalting by crystallization: detection of attomole biomolecules in picoliter buffers by mass spectrometry.

Science.gov (United States)

Gong, Xiaoyun; Xiong, Xingchuang; Wang, Song; Li, Yanyan; Zhang, Sichun; Fang, Xiang; Zhang, Xinrong

2015-10-06

Sensitive detection of biomolecules in small-volume samples by mass spectrometry is, in many cases, challenging because of the use of buffers to maintain the biological activities of proteins and cells. Here, we report a highly effective desalting method for picoliter samples. It was based on the spontaneous separation of biomolecules from salts during crystallization of the salts. After desalting, the biomolecules were deposited in the tip of the quartz pipet because of the evaporation of the solvent. Subsequent detection of the separated biomolecules was achieved using solvent assisted electric field induced desorption/ionization (SAEFIDI) coupled with mass spectrometry. It allowed for direct desorption/ionization of the biomolecules in situ from the tip of the pipet. The organic component in the assistant solvent inhibited the desorption/ionization of salts, thus assured successful detection of biomolecules. Proteins and peptides down to 50 amol were successfully detected using our method even if there were 3 × 10(5) folds more amount of salts in the sample. The concentration and ion species of the salts had little influence on the detection results.

Effect of the sequence data deluge on the performance of methods for detecting protein functional residues.

Science.gov (United States)

Garrido-Martín, Diego; Pazos, Florencio

2018-02-27

The exponential accumulation of new sequences in public databases is expected to improve the performance of all the approaches for predicting protein structural and functional features. Nevertheless, this was never assessed or quantified for some widely used methodologies, such as those aimed at detecting functional sites and functional subfamilies in protein multiple sequence alignments. Using raw protein sequences as only input, these approaches can detect fully conserved positions, as well as those with a family-dependent conservation pattern. Both types of residues are routinely used as predictors of functional sites and, consequently, understanding how the sequence content of the databases affects them is relevant and timely. In this work we evaluate how the growth and change with time in the content of sequence databases affect five sequence-based approaches for detecting functional sites and subfamilies. We do that by recreating historical versions of the multiple sequence alignments that would have been obtained in the past based on the database contents at different time points, covering a period of 20 years. Applying the methods to these historical alignments allows quantifying the temporal variation in their performance. Our results show that the number of families to which these methods can be applied sharply increases with time, while their ability to detect potentially functional residues remains almost constant. These results are informative for the methods' developers and final users, and may have implications in the design of new sequencing initiatives.

Biofouling Removal and Protein Detection Using a Hypersonic Resonator.

Science.gov (United States)

Pan, Shuting; Zhang, Hongxiang; Liu, Wenpeng; Wang, Yanyan; Pang, Wei; Duan, Xuexin

2017-08-25

Nonspecific binding (NSB) is a general issue for surface based biosensors. Various approaches have been developed to prevent or remove the NSBs. However, these approaches either increased the background signals of the sensors or limited to specific transducers interface. In this work, we developed a hydrodynamic approach to selectively remove the NSBs using a microfabricated hypersonic resonator with 2.5 gigahertz (GHz) resonant frequency. The high frequency device facilitates generation of multiple controlled microvortexes which then create cleaning forces at the solid-liquid interfaces. The competitive adhesive and cleaning forces have been investigated using the finite element method (FEM) simulation, identifying the feasibility of the vortex-induced NSB removal. NSB proteins have been selectively removed experimentally both on the surface of the resonator and on other substrates which contact the vortexes. Thus, the developed hydrodynamic approach is believed to be a simple and versatile tool for NSB removal and compatible to many sensor systems. The unique feature of the hypersonic resonator is that it can be used as a gravimetric sensor as well; thus a combined NSB removal and protein detection dual functional biosensor system is developed.

Detecting differential protein expression in large-scale population proteomics

Energy Technology Data Exchange (ETDEWEB)

Ryu, Soyoung; Qian, Weijun; Camp, David G.; Smith, Richard D.; Tompkins, Ronald G.; Davis, Ronald W.; Xiao, Wenzhong

2014-06-17

Mass spectrometry-based high-throughput quantitative proteomics shows great potential in clinical biomarker studies, identifying and quantifying thousands of proteins in biological samples. However, methods are needed to appropriately handle issues/challenges unique to mass spectrometry data in order to detect as many biomarker proteins as possible. One issue is that different mass spectrometry experiments generate quite different total numbers of quantified peptides, which can result in more missing peptide abundances in an experiment with a smaller total number of quantified peptides. Another issue is that the quantification of peptides is sometimes absent, especially for less abundant peptides and such missing values contain the information about the peptide abundance. Here, we propose a Significance Analysis for Large-scale Proteomics Studies (SALPS) that handles missing peptide intensity values caused by the two mechanisms mentioned above. Our model has a robust performance in both simulated data and proteomics data from a large clinical study. Because varying patients’ sample qualities and deviating instrument performances are not avoidable for clinical studies performed over the course of several years, we believe that our approach will be useful to analyze large-scale clinical proteomics data.

Improved Peak Detection and Deconvolution of Native Electrospray Mass Spectra from Large Protein Complexes.

Science.gov (United States)

Lu, Jonathan; Trnka, Michael J; Roh, Soung-Hun; Robinson, Philip J J; Shiau, Carrie; Fujimori, Danica Galonic; Chiu, Wah; Burlingame, Alma L; Guan, Shenheng

2015-12-01

Native electrospray-ionization mass spectrometry (native MS) measures biomolecules under conditions that preserve most aspects of protein tertiary and quaternary structure, enabling direct characterization of large intact protein assemblies. However, native spectra derived from these assemblies are often partially obscured by low signal-to-noise as well as broad peak shapes because of residual solvation and adduction after the electrospray process. The wide peak widths together with the fact that sequential charge state series from highly charged ions are closely spaced means that native spectra containing multiple species often suffer from high degrees of peak overlap or else contain highly interleaved charge envelopes. This situation presents a challenge for peak detection, correct charge state and charge envelope assignment, and ultimately extraction of the relevant underlying mass values of the noncovalent assemblages being investigated. In this report, we describe a comprehensive algorithm developed for addressing peak detection, peak overlap, and charge state assignment in native mass spectra, called PeakSeeker. Overlapped peaks are detected by examination of the second derivative of the raw mass spectrum. Charge state distributions of the molecular species are determined by fitting linear combinations of charge envelopes to the overall experimental mass spectrum. This software is capable of deconvoluting heterogeneous, complex, and noisy native mass spectra of large protein assemblies as demonstrated by analysis of (1) synthetic mononucleosomes containing severely overlapping peaks, (2) an RNA polymerase II/α-amanitin complex with many closely interleaved ion signals, and (3) human TriC complex containing high levels of background noise. Graphical Abstract ᅟ.

Immunohistochemical detection of prion protein in lymphoid tissues of sheep with natural scrapie

NARCIS (Netherlands)

Keulen, van L.J.M.; Schreuder, B.E.C.; Meloen, R.H.; Mooij-Harkes, G.; Vromans, M.E.W.; Langeveld, J.P.M.

1996-01-01

The scrapie-associated form of the prion protein (PrP(Sc)) accumulates in the brain and lymphoid tissues of sheep with scrapie. In order to assess whether detecting PrP(Sc) in lymphoid tissue could he used as a diagnostic test for scrapie, we studied the localization and distribution of PrP(Sc) in

Low-frequency electrical stimulation induces the proliferation and differentiation of peripheral blood stem cells into Schwann cells.

Science.gov (United States)

Gu, Xudong; Fu, Jianming; Bai, Jing; Zhang, Chengwen; Wang, Jing; Pan, Wenping

2015-02-01

Functional recovery after peripheral nerve injury remains a tough problem at present. Specifically, a type of glial cell exists in peripheral nerves that promotes axonal growth and myelin formation and secretes various active substances, such as neurotrophic factors, extracellular matrix and adherence factors. These substances have important significance for the survival, growth and regeneration of nerve fibers. Numerous recent studies have shown that electrical stimulation can increase the number of myelinated nerve fibers. However, whether electrical stimulation acts on neurons or Schwann cells has not been verified in vivo. This study investigates low-frequency electrical stimulation-induced proliferation and differentiation of peripheral blood stem cells into Schwann cells and explores possible mechanisms. Peripheral blood stem cells from Sprague-Dawley rats were primarily cultured. Cells in passage 3 were divided into 4 groups: a low-frequency electrical stimulation group (20 Hz, 100 μs, 3 V), a low-frequency electrical stimulation+PD98059 (blocking the extracellular signal-regulated kinase [ERK] signaling pathway) group, a PD98059 group and a control group (no treatment). After induction, the cells were characterized. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazoliumbromide assay was employed to measure the absorbance values at 570 nm in the 4 groups. A Western blot assay was used to detect the expression of cyclin D1 and cyclin-dependent kinase 4 (CDK4) in each group. No significant difference in cell viability was detected before induction. Peripheral blood stem cells from the 4 groups differentiated into Schwann cells. Phosphorylated ERK 1/2, cyclin D1 and CDK4 protein levels were highest in the low-frequency electrical stimulation group and lowest in the ERK blockage group. Phosphorylated ERK 1/2, cyclin D1 and CDK4 protein levels in the low-frequency electrical stimulation+ERK blockage group were lower than those in the low-frequency electrical

ProBiS-2012: web server and web services for detection of structurally similar binding sites in proteins.

Science.gov (United States)

Konc, Janez; Janezic, Dusanka

2012-07-01

The ProBiS web server is a web server for detection of structurally similar binding sites in the PDB and for local pairwise alignment of protein structures. In this article, we present a new version of the ProBiS web server that is 10 times faster than earlier versions, due to the efficient parallelization of the ProBiS algorithm, which now allows significantly faster comparison of a protein query against the PDB and reduces the calculation time for scanning the entire PDB from hours to minutes. It also features new web services, and an improved user interface. In addition, the new web server is united with the ProBiS-Database and thus provides instant access to pre-calculated protein similarity profiles for over 29 000 non-redundant protein structures. The ProBiS web server is particularly adept at detection of secondary binding sites in proteins. It is freely available at http://probis.cmm.ki.si/old-version, and the new ProBiS web server is at http://probis.cmm.ki.si.

A tosyl-activated magnetic bead cellulose as solid support for sensitive protein detection

Czech Academy of Sciences Publication Activity Database

Yan, J.; Horák, Daniel; Lenfeld, Jiří; Hammond, M.; Kamali-Moghaddam, M.

2013-01-01

Roč. 167, č. 3 (2013), s. 235-240 ISSN 0168-1656 R&D Projects: GA ČR GAP503/10/0664; GA MŠk 7E12054 EU Projects: European Commission(XE) 259796 - DIATOOLS Institutional support: RVO:61389013 Keywords : bead cellulose * magnetic * protein detection Subject RIV: CD - Macromolecular Chemistry Impact factor: 2.884, year: 2013

Detection of dietary DNA, protein, and glyphosate in meat, milk, and eggs.

Science.gov (United States)

Van Eenennaam, A L; Young, A E

2017-07-01

Products such as meat, milk, and eggs from animals that have consumed genetically engineered (GE) feed are not currently subject to mandatory GE labeling requirements. Some voluntary "non-genetically modified organism" labeling has been associated with such products, indicating that the animals were not fed GE crops, as there are no commercialized GE food animals. This review summarizes the available scientific literature on the detection of dietary DNA and protein in animal products and briefly discusses the implications of mandatory GE labeling for products from animals that have consumed GE feed. Because glyphosate is used on some GE crops, the available studies on glyphosate residues in animal products are also reviewed. In GE crops, recombinant DNA (rDNA) makes up a small percentage of the plant's total DNA. The final amount of DNA in food/feed depends on many factors including the variable number and density of cells in the edible parts, the DNA-containing matrix, environmental conditions, and the specific transgenic event. Processing treatments and animals' digestive systems degrade DNA into small fragments. Available reports conclude that endogenous DNA and rDNA are processed in exactly the same way in the gastrointestinal tract and that they account for a very small proportion of food intake by weight. Small pieces of high copy number endogenous plant genes have occasionally been detected in meat and milk. Similarly sized pieces of rDNA have also been identified in meat, primarily fish, although detection is inconsistent. Dietary rDNA fragments have not been detected in chicken or quail eggs or in fresh milk from cows or goats. Collectively, studies have failed to identify full-length endogenous or rDNA transcripts or recombinant proteins in meat, milk, or eggs. Similarly, because mammals do not bioaccumulate glyphosate and it is rapidly excreted, negligible levels of glyphosate in cattle, pig and poultry meat, milk, and eggs have been reported. Despite

Detection of DNA strand breaks in mammalian cells using the radioresistant bacterium PprA protein

International Nuclear Information System (INIS)

Satoh, Katsuya; Wada, Seiichi; Narumi, Issay; Kikuchi, Masahiro; Funayama, Tomoo; Kobayashi, Yasuhiko

2003-01-01

We have previously found that the PprA protein from Deinococcus radiodurans possesses ability to recognize DNA carrying strand breaks. In the present study, we attempted to visualize radiation-induced DNA strand breaks with PprA protein using immunofluorescence technique to elucidate the DNA damage response mechanism in mammalian cultured cells. As a result, colocalization of Cy2 and DAPI fluorescent signals was observed. This observation suggests that DNA strand breaks in the nucleus of CHO-K1 cells were effectively detected using the PprA protein. The amount of DNA strand breaks (integrated density of Cy2 fluorescent signals) was increased with the increase in the radiation dose. (author)

Optimizing FRET-FLIM Labeling Conditions to Detect Nuclear Protein Interactions at Native Expression Levels in Living Arabidopsis Roots

KAUST Repository

Long, Yuchen

2018-05-15

Protein complex formation has been extensively studied using Förster resonance energy transfer (FRET) measured by Fluorescence Lifetime Imaging Microscopy (FLIM). However, implementing this technology to detect protein interactions in living multicellular organism at single-cell resolution and under native condition is still difficult to achieve. Here we describe the optimization of the labeling conditions to detect FRET-FLIM in living plants. This study exemplifies optimization procedure involving the identification of the optimal position for the labels either at the N or C terminal region and the selection of the bright and suitable, fluorescent proteins as donor and acceptor labels for the FRET study. With an effective optimization strategy, we were able to detect the interaction between the stem cell regulators SHORT-ROOT and SCARECROW at endogenous expression levels in the root pole of living Arabidopsis embryos and developing lateral roots by FRET-FLIM. Using this approach we show that the spatial profile of interaction between two transcription factors can be highly modulated in reoccurring and structurally resembling organs, thus providing new information on the dynamic redistribution of nuclear protein complex configurations in different developmental stages. In principle, our optimization procedure for transcription factor complexes is applicable to any biological system.

Detecting coordinated regulation of multi-protein complexes using logic analysis of gene expression

Directory of Open Access Journals (Sweden)

Yeates Todd O

2009-12-01

Full Text Available Abstract Background Many of the functional units in cells are multi-protein complexes such as RNA polymerase, the ribosome, and the proteasome. For such units to work together, one might expect a high level of regulation to enable co-appearance or repression of sets of complexes at the required time. However, this type of coordinated regulation between whole complexes is difficult to detect by existing methods for analyzing mRNA co-expression. We propose a new methodology that is able to detect such higher order relationships. Results We detect coordinated regulation of multiple protein complexes using logic analysis of gene expression data. Specifically, we identify gene triplets composed of genes whose expression profiles are found to be related by various types of logic functions. In order to focus on complexes, we associate the members of a gene triplet with the distinct protein complexes to which they belong. In this way, we identify complexes related by specific kinds of regulatory relationships. For example, we may find that the transcription of complex C is increased only if the transcription of both complex A AND complex B is repressed. We identify hundreds of examples of coordinated regulation among complexes under various stress conditions. Many of these examples involve the ribosome. Some of our examples have been previously identified in the literature, while others are novel. One notable example is the relationship between the transcription of the ribosome, RNA polymerase and mannosyltransferase II, which is involved in N-linked glycan processing in the Golgi. Conclusions The analysis proposed here focuses on relationships among triplets of genes that are not evident when genes are examined in a pairwise fashion as in typical clustering methods. By grouping gene triplets, we are able to decipher coordinated regulation among sets of three complexes. Moreover, using all triplets that involve coordinated regulation with the ribosome

Electric organ discharges and electric images during electrolocation

Science.gov (United States)

Assad, C.; Rasnow, B.; Stoddard, P. K.

1999-01-01

Weakly electric fish use active electrolocation - the generation and detection of electric currents - to explore their surroundings. Although electrosensory systems include some of the most extensively understood circuits in the vertebrate central nervous system, relatively little is known quantitatively about how fish electrolocate objects. We believe a prerequisite to understanding electrolocation and its underlying neural substrates is to quantify and visualize the peripheral electrosensory information measured by the electroreceptors. We have therefore focused on reconstructing both the electric organ discharges (EODs) and the electric images resulting from nearby objects and the fish's exploratory behaviors. Here, we review results from a combination of techniques, including field measurements, numerical and semi-analytical simulations, and video imaging of behaviors. EOD maps are presented and interpreted for six gymnotiform species. They reveal diverse electric field patterns that have significant implications for both the electrosensory and electromotor systems. Our simulations generated predictions of the electric images from nearby objects as well as sequences of electric images during exploratory behaviors. These methods are leading to the identification of image features and computational algorithms that could reliably encode electrosensory information and may help guide electrophysiological experiments exploring the neural basis of electrolocation.

A visual detection of protein content based on titration of moving reaction boundary electrophoresis.

Science.gov (United States)

Wang, Hou-Yu; Guo, Cheng-Ye; Guo, Chen-Gang; Fan, Liu-Yin; Zhang, Lei; Cao, Cheng-Xi

2013-04-24

A visual electrophoretic titration method was firstly developed from the concept of moving reaction boundary (MRB) for protein content analysis. In the developed method, when the voltage was applied, the hydroxide ions in the cathodic vessel moved towards the anode, and neutralized the carboxyl groups of protein immobilized via highly cross-linked polyacrylamide gel (PAG), generating a MRB between the alkali and the immobilized protein. The boundary moving velocity (V(MRB)) was as a function of protein content, and an acid-base indicator was used to denote the boundary displacement. As a proof of concept, standard model proteins and biological samples were chosen for the experiments to study the feasibility of the developed method. The experiments revealed that good linear calibration functions between V(MRB) and protein content (correlation coefficients R>0.98). The experiments further demonstrated the following merits of developed method: (1) weak influence of non-protein nitrogen additives (e.g., melamine) adulterated in protein samples, (2) good agreement with the classic Kjeldahl method (R=0.9945), (3) fast measuring speed in total protein analysis of large samples from the same source, and (4) low limit of detection (0.02-0.15 mg mL(-1) for protein content), good precision (R.S.D. of intra-day less than 1.7% and inter-day less than 2.7%), and high recoveries (105-107%). Crown Copyright © 2013. Published by Elsevier B.V. All rights reserved.

Detection of animal-derived proteins in feedstuffs in Italy: a reproducibility study.

Science.gov (United States)

Ingravalle, Francesco; Abete, Maria Cesarina; Crescio, Maria Ines; Ru, Giuseppe

2007-04-01

Bovine spongiform encephalopathy is a prion disease of ruminants that was first recognized in 1986 in the United Kingdom. Early in the epidemic, it became obvious that the presence of meat and bone meal in feed rations was a common factor in all bovine spongiform encephalopathy cases. The first ban of derived animal proteins in feed was enforced in Europe in 1994 and implemented by Regulation 999/2001 that prohibited the feeding of animal-derived protein to farm animals. The only official method currently accepted by the European Union Commission for test for the presence of animal-derived proteins in feedstuffs is feed microscopy. In Italy, monitoring of feedstuff safety is provided by both the Ministry of Health and the Ministry of Agriculture. The quality of official control, usually assessed by verifying the reproducibility and the accuracy of the testing method, is of fundamental importance for all laboratories and institutions using these results for comparative purposes. The aims of this study were to assess the reproducibility of the official method over all the Italian surveillance network and to provide a model for evaluating the performance of the monitoring system. The accuracy of the identification of the animal class of derived protein detected (avian, mammalian, or aquatic organism) was assessed. The interlaboratory agreement within the overall network reached 0.97 (95% confidence interval of 0.95 to 0.98) for determining the presence or absence of animal-derived proteins (e.g., for mammalian, avian, or aquatic species), and specificity of the identification of the animal class indicated that fish proteins are more easily recognized than are avian or mammalian proteins.

Detection and Elimination of a Potential Fire in Engine and Battery Compartments of Hybrid Electric Vehicles

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Macam S. Dattathreya

2012-01-01

Full Text Available This paper presents a novel fuzzy deterministic noncontroller type (FDNCT system and an FDNCT inference algorithm (FIA. The FDNCT uses fuzzy inputs and produces a deterministic non-fuzzy output. The FDNCT is an extension and alternative for the existing fuzzy singleton inference algorithm. The research described in this paper applies FDNCT to build an architecture for an intelligent system to detect and to eliminate potential fires in the engine and battery compartments of a hybrid electric vehicle. The fuzzy inputs consist of sensor data from the engine and battery compartments, namely, temperature, moisture, and voltage and current of the battery. The system synthesizes the data and detects potential fires, takes actions for eliminating the hazard, and notifies the passengers about the potential fire using an audible alarm. This paper also presents the computer simulation results of the comparison between the FIA and singleton inference algorithms for detecting potential fires and determining the actions for eliminating them.

Amperometric Immunosensor Based on a Protein A/Deposited Gold Nanocrystals Modified Electrode for Carbofuran Detection

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Xia Sun

2011-12-01

Full Text Available In this paper, an amperometric immunosensor modified with protein A/deposited gold nanocrystals (DpAu was developed for the ultrasensitive detection of carbofuran residues. First, DpAu were electrodeposited onto the Au electrode surface to absorb protein A (PA and improve the electrode conductivity. Then PA was dropped onto the surface of DpAu film, used for binding antibody Fc fragments. Next, anti-carbofuran monoclonal antibody was immobilized on the PA modified electrode. Finally, bovine serum albumin (BSA was employed to block the possible remaining active sites avoiding any nonspecific adsorption. The fabrication procedure of the immunosensor was characterized by electrochemical impedance spectroscopy (EIS and cyclic voltammetry (CV, respectively. With the excellent electroconductivity of DpAu and the PA’s oriented immobilization of antibodies, a highly efficient immuno-reaction and detection sensitivity could be achieved. The influences of the electrodeposition time of DpAu, pH of the detection solution and incubation time on the current response of the fabricated immunosensor were investigated. Under optimized conditions, the current response was proportional to the concentration of carbofuran which ranged from 1 to 100 ng/mL and 100 ng/mL to 100 μg/mL. The detection limit was 0.1924 ng/mL. The proposed carbofuran immnuosensor exhibited high specificity, reproducibility, stability and regeneration performance, which may open a new door for ultrasensitive detection of carbofuran residues in vegetables and fruits.

The bee, the flower and the electric field

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Robert Daniel

2016-01-01

Full Text Available Insects use several different senses to forage on flowers, and detect floral cues such as color, shape, pattern, humidity and chemical volatiles. This presentation will present our discovery of a previously unappreciated sensory capacity in bumblebees (Bombus terrestris: the detection of floral electric fields. We show that these floral fields act as informational cues, and that they can be affected by the visit of naturally electrically charged bees. Like visual cues, floral electric fields exhibit variations in pattern and structure, which can be discriminated by bumblebees. We also show that such electric field information contributes to the complex array of floral cues that together improve a pollinator’s memory of floral rewards. Floral electric fields arise from complex interactions with the surrounding atmosphere, an interaction between plants and their environment that not well understood. Because floral electric fields can change within seconds, this new sensory modality - electrostatic field detection- may facilitate rapid and dynamic communication between flowers and their pollinators.

Detection of Side Chain Rearrangements Mediating the Motions of Transmembrane Helices in Molecular Dynamics Simulations of G Protein-Coupled Receptors

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Zied Gaieb

Full Text Available Structure and dynamics are essential elements of protein function. Protein structure is constantly fluctuating and undergoing conformational changes, which are captured by molecular dynamics (MD simulations. We introduce a computational framework that provides a compact representation of the dynamic conformational space of biomolecular simulations. This method presents a systematic approach designed to reduce the large MD simulation spatiotemporal datasets into a manageable set in order to guide our understanding of how protein mechanics emerge from side chain organization and dynamic reorganization. We focus on the detection of side chain interactions that undergo rearrangements mediating global domain motions and vice versa. Side chain rearrangements are extracted from side chain interactions that undergo well-defined abrupt and persistent changes in distance time series using Gaussian mixture models, whereas global domain motions are detected using dynamic cross-correlation. Both side chain rearrangements and global domain motions represent the dynamic components of the protein MD simulation, and are both mapped into a network where they are connected based on their degree of coupling. This method allows for the study of allosteric communication in proteins by mapping out the protein dynamics into an intramolecular network to reduce the large simulation data into a manageable set of communities composed of coupled side chain rearrangements and global domain motions. This computational framework is suitable for the study of tightly packed proteins, such as G protein-coupled receptors, and we present an application on a seven microseconds MD trajectory of CC chemokine receptor 7 (CCR7 bound to its ligand CCL21. Keywords: Molecular dynamics, Change-point detection, Side chain reorganization, Helical domain motion, Intramolecular network, Membrane proteins, GPCR, GPCR computational modeling, GPCR allostery

High-performance fluorescence-encoded magnetic microbeads as microfluidic protein chip supports for AFP detection

Energy Technology Data Exchange (ETDEWEB)

Gong, Xiaoqun [School of Life Sciences, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072 (China); Yan, Huan; Yang, Jiumin [Department of Laboratory Medicine, Tianjin Medical University General Hospital, Tianjin, 300052 (China); Wu, Yudong; Zhang, Jian; Yao, Yingyi [School of Life Sciences, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072 (China); Liu, Ping [Bioscience (Tianjin) Diagnostic Technology CO., LTD, Tianjin, 300300 (China); Wang, Huiquan [Department of Biomedical Engineering, School of Electronics and Information Engineering, Tianjin Polytechnic University, Tianjin, 300387 (China); Hu, Zhidong, E-mail: huzhidong27@163.com [Department of Laboratory Medicine, Tianjin Medical University General Hospital, Tianjin, 300052 (China); Chang, Jin, E-mail: jinchang@tju.edu.cn [School of Life Sciences, Tianjin Engineering Center of Micro-Nano Biomaterials and Detection-Treatment Technology, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072 (China)

2016-10-05

Fluorescence-encoded magnetic microbeads (FEMMs), with the fluorescence encoding ability of quantum dots (QDs) and magnetic enrichment and separation functions of Fe{sub 3}O{sub 4} nanoparticles, have been widely used for multiple biomolecular detection as microfluidic protein chip supports. However, the preparation of FEMMs with long-term fluorescent encoding and immunodetection stability is still a challenge. In this work, we designed a novel high-temperature chemical swelling strategy. The QDs and Fe{sub 3}O{sub 4} nanoparticles were effectively packaged into microbeads via the thermal motion of the polymer chains and the hydrophobic interaction between the nanoparticles and microbeads. The FEMMs obtained a highly uniform fluorescent property and long-term encoding and immunodetection stability and could be quickly magnetically separated and enriched. Then, the QD-encoded magnetic microbeads were applied to alpha fetoprotein (AFP) detection via sandwich immunoreaction. The properties of the encoded microspheres were characterized using a self-designed detecting apparatus, and the target molecular concentration in the sample was also quantified. The results suggested that the high-performance FEMMs have great potential in the field of biomolecular detection. - Graphical abstract: We designed a novel strategy to prepare a kind of high-performance fluorescence-encoded magnetic microbeads as microfluidic protein chip support with long-time fluorescent encoding and immunodetection stability for AFP detection. - Highlights: • A novel strategy combined the high temperature with chemical swelling technology is designed. • Based on hydrophobic interaction and polymer thermal motion, QDs and Fe{sub 3}O{sub 4} were effectively packaged into microbeads. • The fluorescence-encoded magnetic microbeads show long-term fluorescent encoding and immunodetection stability.

Rapid and quantitative detection of C-reactive protein based on quantum dots and immunofiltration assay

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Zhang PF

2015-09-01

Full Text Available Pengfei Zhang,1,* Yan Bao,1,* Mohamed Shehata Draz,2,3,* Huiqi Lu,1 Chang Liu,1 Huanxing Han11Center for Translational Medicine, Changzheng Hospital, Second Military Medical University, Shanghai, People’s Republic of China; 2Zhejiang-California International Nanosystems Institute, Zhejiang University, Hangzhou, Zhejiang, People’s Republic of China; 3Faculty of Science, Tanta University, Tanta, Egypt*These authors contributed equally to this workAbstract: Convenient and rapid immunofiltration assays (IFAs enable on-site “yes” or “no” determination of disease markers. However, traditional IFAs are commonly qualitative or semi-quantitative and are very limited for the efficient testing of samples in field diagnostics. Here, we overcome these limitations by developing a quantum dots (QDs-based fluorescent IFA for the quantitative detection of C-reactive proteins (CRP. CRP, the well-known diagnostic marker for acute viral and bacterial infections, was used as a model analyte to demonstrate performance and sensitivity of our developed QDs-based IFA. QDs capped with both polyethylene glycol (PEG and glutathione were used as fluorescent labels for our IFAs. The presence of the surface PEG layer, which reduced the non-specific protein interactions, in conjunction with the inherent optical properties of QDs, resulted in lower background signal, increased sensitivity, and ability to detect CRP down to 0.79 mg/L with only 5 µL serum sample. In addition, the developed assay is simple, fast and can quantitatively detect CRP with a detection limit up to 200 mg/L. Clinical test results of our QD-based IFA are well correlated with the traditional latex enhance immune-agglutination aggregation. The proposed QD-based fluorescent IFA is very promising, and potentially will be adopted for multiplexed immunoassay and in field point-of-care test.Keywords: C-reactive proteins, point-of-care test, Glutathione capped QDs, PEGylation

Detection of trans–cis flips and peptide-plane flips in protein structures

Energy Technology Data Exchange (ETDEWEB)

Touw, Wouter G., E-mail: wouter.touw@radboudumc.nl [Radboud University Medical Center, Geert Grooteplein-Zuid 26-28, 6525 GA Nijmegen (Netherlands); Joosten, Robbie P. [Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam (Netherlands); Vriend, Gert, E-mail: wouter.touw@radboudumc.nl [Radboud University Medical Center, Geert Grooteplein-Zuid 26-28, 6525 GA Nijmegen (Netherlands)

2015-07-28

A method is presented to detect peptide bonds that need either a trans–cis flip or a peptide-plane flip. A coordinate-based method is presented to detect peptide bonds that need correction either by a peptide-plane flip or by a trans–cis inversion of the peptide bond. When applied to the whole Protein Data Bank, the method predicts 4617 trans–cis flips and many thousands of hitherto unknown peptide-plane flips. A few examples are highlighted for which a correction of the peptide-plane geometry leads to a correction of the understanding of the structure–function relation. All data, including 1088 manually validated cases, are freely available and the method is available from a web server, a web-service interface and through WHAT-CHECK.

Detection of trans–cis flips and peptide-plane flips in protein structures

International Nuclear Information System (INIS)

Touw, Wouter G.; Joosten, Robbie P.; Vriend, Gert

2015-01-01

A method is presented to detect peptide bonds that need either a trans–cis flip or a peptide-plane flip. A coordinate-based method is presented to detect peptide bonds that need correction either by a peptide-plane flip or by a trans–cis inversion of the peptide bond. When applied to the whole Protein Data Bank, the method predicts 4617 trans–cis flips and many thousands of hitherto unknown peptide-plane flips. A few examples are highlighted for which a correction of the peptide-plane geometry leads to a correction of the understanding of the structure–function relation. All data, including 1088 manually validated cases, are freely available and the method is available from a web server, a web-service interface and through WHAT-CHECK

Detection of anti neutrophil antibodies by radio-iodinated protein A binding test

International Nuclear Information System (INIS)

Cartron, J.; Muller, J.Y.; Tchernia, G.; Paule, B.; Varet, B.

1983-01-01

The granulocyte associated IgG in normal and neutropenic subjects has been determined by a direct quantitative assay using radiolabeled staphylococcal protein A. This assay allows to postulate an immunological mechanism to explain the neutropenia in 19 cases of neutropenias associated with malfunctions of the immune system and in 4 cases of idiopathic neutropenias. Discussed in this report is the possible interaction of immune complexes bound in vivo to the granulocytes. By an immunofluorescence test, it has been possible to detect IgG or IgM antibodies in only 52% of patients with a positive direct assay. The determination of granulocyte-associated IgG is therefore a better indicator for defining an auto-immune neutropenia than the detection of free antibodies in the sera [fr

Proximity hybridization-mediated isothermal exponential amplification for ultrasensitive electrochemical protein detection

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Yu Y

2017-08-01

Full Text Available Yanyan Yu, Gaoxing Su, Hongyan Zhu, Qing Zhu, Yong Chen, Bohui Xu, Yuqin Li, Wei Zhang School of Pharmacy, Nantong University, Nantong, People’s Republic of China Abstract: In this study, we fabricated a novel electrochemical biosensing platform on the basis of target-triggered proximity hybridization-mediated isothermal exponential amplification reaction (EXPAR for ultrasensitive protein analysis. Through rational design, the aptamers for protein recognition were integrated within two DNA probes. Via proximity hybridization principle, the affinity protein-binding event was converted into DNA assembly process. The recognition of protein by aptamers can trigger the strand displacement through the increase of the local concentrations of the involved probes. As a consequence, the output DNA was displaced, which can hybridize with the duplex probes immobilized on the electrode surface subsequently, leading to the initiation of the EXPAR as well as the cleavage of duplex probes. Each cleavage will release the gold nanoparticles (AuNPs binding sequence. With the modification of G-quadruplex sequence, electrochemical signals were yielded by the AuNPs through oxidizing 3,3',5,5'-tetramethylbenzidine in the presence of H2O2. The study we proposed exhibited high sensitivity toward platelet-derived growth factor BB (PDGF-BB with the detection limit of 52 fM. And, this method also showed great selectivity among the PDGF isoforms and performed well in spiked human serum samples. Keywords: electrochemical biosensor, proximity hybridization, PDGF-BB, isothermal exponential amplification, G-quadruplex 

Urine protein electrophoresis test

Science.gov (United States)

Urine protein electrophoresis; UPEP; Multiple myeloma - UPEP; Waldenström macroglobulinemia - UPEP; Amyloidosis - UPEP ... special paper and apply an electric current. The proteins move and form visible bands. These reveal the ...

The application of diode laser colorimetry coupled with fiber optic dipping probe for quantitative detection of a protein

International Nuclear Information System (INIS)

Kim, Sung Ho; Yoo, Jong Shin

1996-01-01

The in-situ, simple and inexpensive analysis of protein was achieved by the portable diode laser absorption spectrometry, which consisted of visible diode laser, photodiode, optical fiber and dipping probe. It gives comparable detection limit to the use of conventional UV/Vis spectrometer for the determination of protein by Lowry method.

Gastric clearance of alpha-1-antitrypsin under cimetidine perfusion. New test to detect protein-losing gastropathy

International Nuclear Information System (INIS)

Florent, C.; Vidon, N.; Flourie, B.; Carmantrand, A.; Zerbani, A.; Maurel, M.; Bernier, J.J.

1986-01-01

Gastric losses of plasma are usually measured with radiolabeled macromolecules. This method is expensive and cumbersome. Direct measurement of exudated plasma proteins are ineffective since proteins are denaturated by acidic gastric juice and pepsin. It was recently shown that albumin measurement after immediate neutralization allowed detection of gastric protein losses, but this method is quite complex and time consuming. We studied alpha 1-antitrypsin and 51Cr-labeled protein clearance in gastric juice during normal saline and cimetidine (1.5 mg/kg/hr) infusion in six healthy volunteers and six patients with exudative gastropathy. alpha 1-Antitrypsin was measurable in all samples during cimetidine infusion: alpha 1-AT and 51Cr losses were significantly correlated (P less than 0.001). The upper limit of gastric alpha 1-AT clearance in controls was 0.86 ml/hr (mean + 2 SD). Using this value, there was no overlapping between patients and controls. The upper limit of 51Cr test was 1.87 ml/hr (mean + 2 SD) in controls but gastric clearance of 51Cr was below this value in one patient. This suggests that the measurement of alpha 1-AT gastric clearance during cimetidine perfusion is a good test to detect an exudative gastropathy. This test is inexpensive and lasts only 3 hr