Fretting wear simulation of press-fitted shaft with finite element analysis and influence function method

Energy Technology Data Exchange (ETDEWEB)

Lee, Dong Hyong; Kwon, Seok Jin [Korea Railroad Research Institute, Uiwang (Korea, Republic of); Choi, Jae Boong; Kim, Young Jin [Sungkyunkwan University, Suwon (Korea, Republic of)

2008-01-15

In this paper the fretting wear of press-fitted specimens subjected to a cyclic bending load was simulated using finite element analysis and numerical method. The amount of microslip and contact variable at press-fitted and bending load condition in a press-fitted shaft was analysed by applying finite element method. With the finite element analysis result, a numerical approach was applied to predict fretting wear based on modified Archard's equation and updating the change of contact pressure caused by local wear with influence function method. The predicted wear profiles of press-fitted specimens at the contact edge wear compared with the experimental results obtained by rotating bending fatigue tests. It is shown that the depth of fretting wear by repeated slip between shaft and boss reaches the maximum value at the contact edge. The initial surface profile is continuously changed by the wear at the contact edge, and then the corresponding contact variables are redistributed. The work establishes a basis for numerical simulation of fretting wear on press fits.

Fretting wear simulation of press-fitted shaft with finite element analysis and influence function method

International Nuclear Information System (INIS)

Lee, Dong Hyong; Kwon, Seok Jin; Choi, Jae Boong; Kim, Young Jin

2008-01-01

In this paper the fretting wear of press-fitted specimens subjected to a cyclic bending load was simulated using finite element analysis and numerical method. The amount of microslip and contact variable at press-fitted and bending load condition in a press-fitted shaft was analysed by applying finite element method. With the finite element analysis result, a numerical approach was applied to predict fretting wear based on modified Archard's equation and updating the change of contact pressure caused by local wear with influence function method. The predicted wear profiles of press-fitted specimens at the contact edge wear compared with the experimental results obtained by rotating bending fatigue tests. It is shown that the depth of fretting wear by repeated slip between shaft and boss reaches the maximum value at the contact edge. The initial surface profile is continuously changed by the wear at the contact edge, and then the corresponding contact variables are redistributed. The work establishes a basis for numerical simulation of fretting wear on press fits

Roughness Effects on Fretting Fatigue

Science.gov (United States)

Yue, Tongyan; Abdel Wahab, Magd

2017-05-01

Fretting is a small oscillatory relative motion between two normal loaded contact surfaces. It may cause fretting fatigue, fretting wear and/or fretting corrosion damage depending on various fretting couples and working conditions. Fretting fatigue usually occurs at partial slip condition, and results in catastrophic failure at the stress levels below the fatigue limit of the material. Many parameters may affect fretting behaviour, including the applied normal load and displacement, material properties, roughness of the contact surfaces, frequency, etc. Since fretting damage is undesirable due to contacting, the effect of rough contact surfaces on fretting damage has been studied by many researchers. Experimental method on this topic is usually focusing on rough surface effects by finishing treatment and random rough surface effects in order to increase fretting fatigue life. However, most of numerical models on roughness are based on random surface. This paper reviewed both experimental and numerical methodology on the rough surface effects on fretting fatigue.

Quantitative FLIM-FRET Microscopy to Monitor Nanoscale Chromatin Compaction In Vivo Reveals Structural Roles of Condensin Complexes

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David Llères

2017-02-01

Full Text Available How metazoan genomes are structured at the nanoscale in living cells and tissues remains unknown. Here, we adapted a quantitative FRET (Förster resonance energy transfer-based fluorescence lifetime imaging microscopy (FLIM approach to assay nanoscale chromatin compaction in living organisms. Caenorhabditis elegans was chosen as a model system. By measuring FRET between histone-tagged fluorescent proteins, we visualized distinct chromosomal regions and quantified the different levels of nanoscale compaction in meiotic cells. Using RNAi and repetitive extrachromosomal array approaches, we defined the heterochromatin state and showed that its architecture presents a nanoscale-compacted organization controlled by Heterochromatin Protein-1 (HP1 and SETDB1 H3-lysine-9 methyltransferase homologs in vivo. Next, we functionally explored condensin complexes. We found that condensin I and condensin II are essential for heterochromatin compaction and that condensin I additionally controls lowly compacted regions. Our data show that, in living animals, nanoscale chromatin compaction is controlled not only by histone modifiers and readers but also by condensin complexes.

Lateral distribution of NBD-PC fluorescent lipid analogs in membranes probed by molecular dynamics-assisted analysis of Förster Resonance Energy Transfer (FRET) and fluorescence quenching.

Science.gov (United States)

Loura, Luís M S

2012-11-08

Förster resonance energy transfer (FRET) is a powerful tool used for many problems in membrane biophysics, including characterization of the lateral distribution of lipid components and other species of interest. However, quantitative analysis of FRET data with a topological model requires adequate choices for the values of several input parameters, some of which are difficult to obtain experimentally in an independent manner. For this purpose, atomistic molecular dynamics (MD) simulations can be potentially useful as they provide direct detailed information on transverse probe localization, relative probe orientation, and membrane surface area, all of which are required for analysis of FRET data. This is illustrated here for the FRET pairs involving 1,6-diphenylhexatriene (DPH) as donor and either 1-palmitoyl,2-(6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] hexanoyl)- sn-glycero-3-phosphocholine (C6-NBD-PC) or 1-palmitoyl,2-(12-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]dodecanoyl)-sn-glycero-3-phosphocholine (C12-NBD-PC) as acceptors, in fluid vesicles of 1,2-dipalmitoyl-sn-3-glycerophosphocholine (DPPC, 50 °C). Incorporation of results from MD simulations improves the statistical quality of model fitting to the experimental FRET data. Furthermore, the decay of DPH in the presence of moderate amounts of C12-NBD-PC (>0.4 mol%) is consistent with non-random lateral distribution of the latter, at variance with C6-NBD-PC, for which aggregation is ruled out up to 2.5 mol% concentration. These conclusions are supported by analysis of NBD-PC fluorescence self-quenching. Implications regarding the relative utility of these probes in membrane studies are discussed.

Prediction of fretting fatigue behavior under elastic-plastic conditions

International Nuclear Information System (INIS)

Shin, Ki Su

2009-01-01

Fretting fatigue generally leads to the degradation of the fatigue strength of a material due to cyclic micro-slip between two contacting materials. Fretting fatigue is regarded as an important issue in designing aerospace structures. While many studies have evaluated fretting fatigue behavior under elastic deformation conditions, few have focused on fretting fatigue behavior under elastic-plastic deformation conditions, especially the crack orientation and fatigue life prediction for Ti-6Al-4V. The primary goal of this study was to characterize the fretting fatigue crack initiation behavior in the presence of plasticity. Experimental tests were performed using pad configurations involving elastic-plastic deformations. To calculate stress distributions under elastic-plastic fretting fatigue conditions, FEA was also performed. Several parametric approaches were used to predict fretting fatigue life along with stress distribution resulting from FEA. However, those parameters using surface stresses were unable to establish an equivalence between elastic fretting fatigue data and elastic-plastic fretting fatigue data. Based on this observation, the critical distance methods, which are commonly used in notch analysis, were applied to the fretting fatigue problem. In conclusion, the effective strain range method when used in conjunction with the SMSSR parameter showed a good correlation of data points between the pad configurations involving elastic and elastic plastic deformations

Application of FRET probes in the analysis of neuronal plasticity

Directory of Open Access Journals (Sweden)

Yoshibumi eUeda

2013-10-01

Full Text Available Breakthroughs in imaging techniques and optical probes in recent years have revolutionized the field of life sciences in ways that traditional methods could never match. The spatial and temporal regulation of molecular events can now be studied with great precision. There have been several key discoveries that have made this possible. Since GFP was cloned in 1992, it has become the dominant tracer of proteins in living cells. Then the evolution of color variants of GFP opened the door to the application of Förster resonance energy transfer (FRET, which is now widely recognized as a powerful tool to study complicated signal transduction events and interactions between molecules. Employment of fluorescent lifetime imaging microscopy (FLIM allows the precise detection of FRET in small subcellular structures such as dendritic spines. In this review, we provide an overview of the basic and practical aspects of FRET imaging and discuss how different FRET probes have revealed insights into the molecular mechanisms of synaptic plasticity and enabled visualization of neuronal network activity both in vitro and in vivo.

N-way FRET microscopy of multiple protein-protein interactions in live cells.

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Adam D Hoppe

Full Text Available Fluorescence Resonance Energy Transfer (FRET microscopy has emerged as a powerful tool to visualize nanoscale protein-protein interactions while capturing their microscale organization and millisecond dynamics. Recently, FRET microscopy was extended to imaging of multiple donor-acceptor pairs, thereby enabling visualization of multiple biochemical events within a single living cell. These methods require numerous equations that must be defined on a case-by-case basis. Here, we present a universal multispectral microscopy method (N-Way FRET to enable quantitative imaging for any number of interacting and non-interacting FRET pairs. This approach redefines linear unmixing to incorporate the excitation and emission couplings created by FRET, which cannot be accounted for in conventional linear unmixing. Experiments on a three-fluorophore system using blue, yellow and red fluorescent proteins validate the method in living cells. In addition, we propose a simple linear algebra scheme for error propagation from input data to estimate the uncertainty in the computed FRET images. We demonstrate the strength of this approach by monitoring the oligomerization of three FP-tagged HIV Gag proteins whose tight association in the viral capsid is readily observed. Replacement of one FP-Gag molecule with a lipid raft-targeted FP allowed direct observation of Gag oligomerization with no association between FP-Gag and raft-targeted FP. The N-Way FRET method provides a new toolbox for capturing multiple molecular processes with high spatial and temporal resolution in living cells.

Standard guide for fretting fatigue testing

CERN Document Server

American Society for Testing and Materials. Philadelphia

2010-01-01

1.1 This guide defines terminology and covers general requirements for conducting fretting fatigue tests and reporting the results. It describes the general types of fretting fatigue tests and provides some suggestions on developing and conducting fretting fatigue test programs. 1.2 Fretting fatigue tests are designed to determine the effects of mechanical and environmental parameters on the fretting fatigue behavior of metallic materials. This guide is not intended to establish preference of one apparatus or specimen design over others, but will establish guidelines for adherence in the design, calibration, and use of fretting fatigue apparatus and recommend the means to collect, record, and reporting of the data. 1.3 The number of cycles to form a fretting fatigue crack is dependent on both the material of the fatigue specimen and fretting pad, the geometry of contact between the two, and the method by which the loading and displacement are imposed. Similar to wear behavior of materials, it is important t...

Evaluation of surface characteristics under fretting of electrical contacts: Removal behaviour of hot dipped tin coating

International Nuclear Information System (INIS)

Park, Young Woo; Ramesh Bapu, G.N.K.; Lee, Kang Yong

2009-01-01

The fretting corrosion behaviour of hot dipped tin coating is investigated at low fretting cycles at ±25 μm displacement amplitude, 0.5N normal load, 3 Hz frequency, 45-50% relative humidity, and 25 ± 1 deg. C temperature. The typical characteristics of the change in contact resistance with fretting cycles are explained. The fretted surface is examined using laser scanning microscope, scanning electron microscope and energy dispersive X-ray analysis to assess the surface profile, extent of fretting damage, extent of oxidation and elemental distribution across the contact zone. The interdependence of extent of wear and oxidation increases the complexity of the fretting corrosion behaviour of tin coating. The variation of contact resistance clearly revealed the fretting of tin coating from 50 to 1200 cycles and the fretting of the substrate above 1200 cycles. The observed low and stable contact resistance region and the fluctuating resistance region at various fretting cycles are explained and substantiated with Scanning electron microscopy (SEM), laser scanning microscope (LSM) and energy dispersive analysis of X-rays (EDAX) analysis results of the fretted surface.

Advanced non-linear flow-induced vibration and fretting-wear analysis capabilities

Energy Technology Data Exchange (ETDEWEB)

Toorani, M.; Pan, L.; Li, R.; Idvorian, N. [Babcock and Wilcox Canada Ltd., Cambridge, Ontario (Canada); Vincent, B.

2009-07-01

Fretting wear is a potentially significant degradation mechanism in nuclear steam generators and other shell and tube heat transfer equipment as well. This paper presents an overview of the recently developed code FIVDYNA which is used for the non-linear flow-induced vibration and fretting wear analysis for operating steam generators (OTSG and RSG) and shell-and-tube heat exchangers. FIVDYNA is a non-linear time-history Flow-Induced Vibration (FIV) analysis computer program that has been developed by Babcock and Wilcox Canada to advance the understanding of tube vibration and tube to tube-support interaction. In addition to the dynamic fluid induced forces the program takes into account other tube static forces due to axial and lateral tube preload and thermal interaction loads. The program is capable of predicting the location where the fretting wear is most likely to occur and its magnitude taking into account the support geometry including gaps. FIVDYNA uses the general purpose finite element computer code ABAQUS as its solver. Using ABAQUS gives the user the flexibility to add additional forces to the tube ranging from tube preloads and the support offsets to thermal loads. The forces currently being modeled in FIVDYNA are the random turbulence, steady drag force, fluid-elastic forces, support offset and pre-strain force (axial loads). This program models the vibration of tubes and calculates the structural dynamic characteristics, and interaction forces between the tube and the tube supports. These interaction forces are then used to calculate the work rate at the support and eventually the predicted depth of wear scar on the tube. A very good agreement is found with experiments and also other computer codes. (author)

Understanding and modeling Förster-type resonance energy transfer (FRET) introduction to FRET

CERN Document Server

Govorov, Alexander; Demir, Hilmi Volkan

2016-01-01

This Brief presents a historical overview of the Förster-type nonradiative energy transfer and a compilation of important progress in FRET research, starting from Förster until today, along with a summary of the current state-of-the-art. Here the objective is to provide the reader with a complete account of important milestones in FRET studies and FRET applications as well as a picture of the current status.

FRET Sensor for Erythrosine Dye Based on Organic Nanoparticles: Application to Analysis of Food Stuff.

Science.gov (United States)

Mahajan, Prasad G; Bhopate, Dhanaji P; Kolekar, Govind B; Patil, Shivajirao R

2016-07-01

fluorescent organic nanoprobe developed for the detection of erythrosine (ETS) food dye in aqueous medium based on fluorescence resonance energy transfer (FRET). The FRET process between donor (nanoparticles) and acceptor (ETS dye) arises due to oppositely charge attraction through hydrophobic interactions. The proposed method was successfully applied to quantitative determination of ETS dye in food stuff sample collected from local market.

EXPERIMENTAL INVESTIGTION OF THE FRETTING PHENOMENON

Directory of Open Access Journals (Sweden)

Ştefan GHIMISI

2015-12-01

Full Text Available Fretting is now fully identified as a small amplitude oscilatory motion which induces a harmonic tangential force between two surfaces in contact.It is related to three main loadings, i.e. fretting-wear, fretting-fatigue and fretting corrosion.Fretting regimes were first mapped by Vingsbo. In a similar way, three fretting regimes will be considered: stick regime,slip regime and mixed regime. The mixed regime was made up of initial gross slip followed by partial slip condition after a few hundred cycles. Obviously the partial slip transition develops the highest stress levels which can induce fatigue crack nucleation depending on the fatigue properties of the two contacting first bodies. Therefore prediction of the frontier between partial slip and gross slip is required.

Fluorescence lifetime FRET imaging of receptor-ligand complexes in tumor cells in vitro and in vivo

Science.gov (United States)

Rudkouskaya, Alena; Sinsuebphon, Nattawut; Intes, Xavier; Mazurkiewicz, Joseph E.; Barroso, Margarida

2017-02-01

To guide the development of targeted therapies with improved efficacy and accelerated clinical acceptance, novel imaging methodologies need to be established. Toward this goal, fluorescence lifetime Förster resonance energy transfer (FLIM-FRET) imaging assays capitalize on the ability of antibodies or protein ligands to bind dimerized membrane bound receptors to measure their target engagement levels in cancer cells. Conventional FLIM FRET microscopy has been widely applied at visible wavelengths to detect protein-protein interactions in vitro. However, operation at these wavelengths restricts imaging quality and ability to quantitate lifetime changes in in vivo small animal optical imaging due to high auto-fluorescence and light scattering. Here, we have analyzed the uptake of iron-bound transferrin (Tf) probes into human breast cancer cells using FLIM-FRET microscopy in the visible and near-infrared (NIR) range. The development of NIR FLIM FRET microscopy allows for the use of quantitative lifetime-based molecular assays to measure drug-target engagement levels at multiple scales: from in vitro microscopy to in vivo small animal optical imaging (macroscopy). This novel approach can be extended to other receptors, currently targeted in oncology. Hence, lifetime-based molecular imaging can find numerous applications in drug delivery and targeted therapy assessment and optimization.

Maximum likelihood-based analysis of photon arrival trajectories in single-molecule FRET

Energy Technology Data Exchange (ETDEWEB)

Waligorska, Marta [Adam Mickiewicz University, Faculty of Chemistry, Grunwaldzka 6, 60-780 Poznan (Poland); Molski, Andrzej, E-mail: amolski@amu.edu.pl [Adam Mickiewicz University, Faculty of Chemistry, Grunwaldzka 6, 60-780 Poznan (Poland)

2012-07-25

Highlights: Black-Right-Pointing-Pointer We study model selection and parameter recovery from single-molecule FRET experiments. Black-Right-Pointing-Pointer We examine the maximum likelihood-based analysis of two-color photon trajectories. Black-Right-Pointing-Pointer The number of observed photons determines the performance of the method. Black-Right-Pointing-Pointer For long trajectories, one can extract mean dwell times that are comparable to inter-photon times. -- Abstract: When two fluorophores (donor and acceptor) are attached to an immobilized biomolecule, anti-correlated fluctuations of the donor and acceptor fluorescence caused by Foerster resonance energy transfer (FRET) report on the conformational kinetics of the molecule. Here we assess the maximum likelihood-based analysis of donor and acceptor photon arrival trajectories as a method for extracting the conformational kinetics. Using computer generated data we quantify the accuracy and precision of parameter estimates and the efficiency of the Akaike information criterion (AIC) and the Bayesian information criterion (BIC) in selecting the true kinetic model. We find that the number of observed photons is the key parameter determining parameter estimation and model selection. For long trajectories, one can extract mean dwell times that are comparable to inter-photon times.

A Toolbox of Genetically Encoded FRET-Based Biosensors for Rapid l-Lysine Analysis

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Victoria Steffen

2016-09-01

Full Text Available Background: The fast development of microbial production strains for basic and fine chemicals is increasingly carried out in small scale cultivation systems to allow for higher throughput. Such parallelized systems create a need for new rapid online detection systems to quantify the respective target compound. In this regard, biosensors, especially genetically encoded Förster resonance energy transfer (FRET-based biosensors, offer tremendous opportunities. As a proof-of-concept, we have created a toolbox of FRET-based biosensors for the ratiometric determination of l-lysine in fermentation broth. Methods: The sensor toolbox was constructed based on a sensor that consists of an optimized central lysine-/arginine-/ornithine-binding protein (LAO-BP flanked by two fluorescent proteins (enhanced cyan fluorescent protein (ECFP, Citrine. Further sensor variants with altered affinity and sensitivity were obtained by circular permutation of the binding protein as well as the introduction of flexible and rigid linkers between the fluorescent proteins and the LAO-BP, respectively. Results: The sensor prototype was applied to monitor the extracellular l-lysine concentration of the l-lysine producing Corynebacterium glutamicum (C. glutamicum strain DM1933 in a BioLector® microscale cultivation device. The results matched well with data obtained by HPLC analysis and the Ninhydrin assay, demonstrating the high potential of FRET-based biosensors for high-throughput microbial bioprocess optimization.

FRET two-hybrid assay by linearly fitting FRET efficiency to concentration ratio between acceptor and donor

Science.gov (United States)

Du, Mengyan; Yang, Fangfang; Mai, Zihao; Qu, Wenfeng; Lin, Fangrui; Wei, Lichun; Chen, Tongsheng

2018-04-01

We here introduce a fluorescence resonance energy transfer (FRET) two-hybrid assay method to measure the maximal donor(D)- and acceptor(A)-centric FRET efficiency (ED,max and EA,max) of the D-A complex and its stoichiometry by linearly fitting the donor-centric FRET efficiency (ED) to the acceptor-to-donor concentration ratio (RC) and acceptor-centric FRET efficiency (EA) to 1/RC, respectively. We performed this method on a wide-field fluorescence microscope for living HepG2 cells co-expressing FRET tandem constructs and free donor/acceptor and obtained correct ED, EA, and stoichiometry values of those tandem constructs. Evaluation on the binding of Bad with Bcl-XL in Hela cells showed that Bad interacted strongly with Bcl-XL to form a Bad-Bcl-XL complex on mitochondria, and one Bad interacted mainly with one Bcl-XL molecule in healthy cells, while with multiple (maybe 2) Bcl-XL molecules in apoptotic cells.

Stuy on Fatigue Life of Aluminum Alloy Considering Fretting

Science.gov (United States)

Yang, Maosheng; Zhao, Hongqiang; Wang, Yunxiang; Chen, Xiaofei; Fan, Jiali

2018-01-01

To study the influence of fretting on Aluminum Alloy, a global finite element model considering fretting was performed using the commercial code ABAQUS. With which a new model for predicting fretting fatigue life has been presented based on friction work. The rationality and effectiveness of the model were validated according to the contrast of experiment life and predicting life. At last influence factor on fretting fatigue life of aerial aluminum alloy was investigated with the model. The results revealed that fretting fatigue life decreased monotonously with the increasing of normal load and then became constant at higher pressures. At low normal load, fretting fatigue life was found to increase with increase in the pad radius. At high normal load, however, the fretting fatigue life remained almost unchanged with changes in the fretting pad radius. The bulk stress amplitude had the dominant effect on fretting fatigue life. The fretting fatigue life diminished as the bulk stress amplitude increased.

Les vicissitudes du fret ferroviaire

OpenAIRE

DABLANC, L

2010-01-01

Dans beaucoup de pays européens, et plus encore en Amérique du Nord et en Asie, le transport de marchandises par le train a augmenté depuis dix ans. Cette activité réduit la part des marchandises acheminées par la route et contribue ainsi au développement durable : un camion émet 8 à 30 fois plus de dioxyde de carbone que le train, pour une distance et une quantité transportée équivalentes. Pourtant, la France a raté ce renouveau. Filiale du groupe public SNCF, la Société Fret SNCF, qui assur...

Theoretical-experimental analysis of the fretting/impact wear in fuel rods

International Nuclear Information System (INIS)

Pecos, Luis F.

2001-01-01

Nuclear power plant fuel elements are subjected to flow induced vibrations. A consequence of these vibrations is impact/fretting wear in fuel rods or sliding shoes. Because of the difficulties to assert the mechanism of impact/fretting wear phenomenon it is necessary to use semiempirical formulations in order to predict the wear rate of the components. The results of a series of experiments with Zr-4 specimens are presented in this work. A parameter called 'work-rate' was used to normalize the wear rates and interpret the results in terms of wear coefficient. (author) [es

Chemical synthesis of dual labeled proteins via differently protected alkynes enables intramolecular FRET analysis.

Science.gov (United States)

Hayashi, Gosuke; Kamo, Naoki; Okamoto, Akimitsu

2017-05-30

We report a novel method for multisite protein conjugation by setting differently silyl-protected alkynes as conjugation handles, which can remain intact through the whole synthetic procedure and provide sequential and orthogonal conjugation. This strategy enables efficient preparation of a dual dye-labeled protein and structural analysis via an intramolecular FRET mechanism.

FRET analysis of CP12 structural interplay by GAPDH and PRK.

Science.gov (United States)

Moparthi, Satish Babu; Thieulin-Pardo, Gabriel; de Torres, Juan; Ghenuche, Petru; Gontero, Brigitte; Wenger, Jérôme

2015-03-13

CP12 is an intrinsically disordered protein playing a key role in the regulation of the Benson-Calvin cycle. Due to the high intrinsic flexibility of CP12, it is essential to consider its structural modulation induced upon binding to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) enzymes. Here, we report for the first time detailed structural modulation about the wild-type CP12 and its site-specific N-terminal and C-terminal disulfide bridge mutants upon interaction with GAPDH and PRK by Förster resonance energy transfer (FRET). Our results indicate an increase in CP12 compactness when the complex is formed with GAPDH or PRK. In addition, the distributions in FRET histograms show the elasticity and conformational flexibility of CP12 in all supra molecular complexes. Contrarily to previous beliefs, our FRET results importantly reveal that both N-terminal and C-terminal site-specific CP12 mutants are able to form the monomeric (GAPDH-CP12-PRK) complex. Copyright © 2015 Elsevier Inc. All rights reserved.

Fluorescence Lifetime Readouts of Troponin-C-Based Calcium FRET Sensors: A Quantitative Comparison of CFP and mTFP1 as Donor Fluorophores

Science.gov (United States)

Laine, Romain; Stuckey, Daniel W.; Manning, Hugh; Warren, Sean C.; Kennedy, Gordon; Carling, David

2012-01-01

We have compared the performance of two Troponin-C-based calcium FRET sensors using fluorescence lifetime read-outs. The first sensor, TN-L15, consists of a Troponin-C fragment inserted between CFP and Citrine while the second sensor, called mTFP-TnC-Cit, was realized by replacing CFP in TN-L15 with monomeric Teal Fluorescent Protein (mTFP1). Using cytosol preparations of transiently transfected mammalian cells, we have measured the fluorescence decay profiles of these sensors at controlled concentrations of calcium using time-correlated single photon counting. These data were fitted to discrete exponential decay models using global analysis to determine the FRET efficiency, fraction of donor molecules undergoing FRET and calcium affinity of these sensors. We have also studied the decay profiles of the donor fluorescent proteins alone and determined the sensitivity of the donor lifetime to temperature and emission wavelength. Live-cell fluorescence lifetime imaging (FLIM) of HEK293T cells expressing each of these sensors was also undertaken. We confirmed that donor fluorescence of mTFP-TnC-Cit fits well to a two-component decay model, while the TN-L15 lifetime data was best fitted to a constrained four-component model, which was supported by phasor analysis of the measured lifetime data. If the constrained global fitting is employed, the TN-L15 sensor can provide a larger dynamic range of lifetime readout than the mTFP-TnC-Cit sensor but the CFP donor is significantly more sensitive to changes in temperature and emission wavelength compared to mTFP and, while the mTFP-TnC-Cit solution phase data broadly agreed with measurements in live cells, this was not the case for the TN-L15 sensor. Our titration experiment also indicates that a similar precision in determination of calcium concentration can be achieved with both FRET biosensors when fitting a single exponential donor fluorescence decay model to the fluorescence decay profiles. We therefore suggest that m

Direct Assessment of the Effect of the Gly380Arg Achondroplasia Mutation on FGFR3 Dimerization Using Quantitative Imaging FRET

Science.gov (United States)

Placone, Jesse; Hristova, Kalina

2012-01-01

The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH), the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase in FGFR3 dimerization due to the ACH mutation. The data are consistent with the idea that the ACH mutation causes a structural change which affects both the stability and the activity of FGFR3 dimers in the absence of ligand. PMID:23056398

Partially reduced graphene oxide based FRET on fiber-optic interferometer for biochemical detection.

Science.gov (United States)

Yao, B C; Wu, Y; Yu, C B; He, J R; Rao, Y J; Gong, Y; Fu, F; Chen, Y F; Li, Y R

2016-03-24

Fluorescent resonance energy transfer (FRET) with naturally exceptional selectivity is a powerful technique and widely used in chemical and biomedical analysis. However, it is still challenging for conventional FRET to perform as a high sensitivity compact sensor. Here we propose a novel 'FRET on Fiber' concept, in which a partially reduced graphene oxide (prGO) film is deposited on a fiber-optic modal interferometer, acting as both the fluorescent quencher for the FRET and the sensitive cladding for optical phase measurement due to refractive index changes in biochemical detection. The target analytes induced fluorescence recovery with good selectivity and optical phase shift with high sensitivity are measured simultaneously. The functionalized prGO film coated on the fiber-optic interferometer shows high sensitivities for the detections of metal ion, dopamine and single-stranded DNA (ssDNA), with detection limits of 1.2 nM, 1.3 μM and 1 pM, respectively. Such a prGO based 'FRET on fiber' configuration, bridging the FRET and the fiber-optic sensing technology, may serve as a platform for the realization of series of integrated 'FRET on Fiber' sensors for on-line environmental, chemical, and biomedical detection, with excellent compactness, high sensitivity, good selectivity and fast response.

Applicability of out-of-pile fretting wear tests to in-reactor fretting wear-induced failure time prediction

Science.gov (United States)

Kim, Kyu-Tae

2013-02-01

In order to investigate whether or not the grid-to-rod fretting wear-induced fuel failure will occur for newly developed spacer grid spring designs for the fuel lifetime, out-of-pile fretting wear tests with one or two fuel assemblies are to be performed. In this study, the out-of-pile fretting wear tests were performed in order to compare the potential for wear-induced fuel failure in two newly-developed, Korean PWR spacer grid designs. Lasting 20 days, the tests simulated maximum grid-to-rod gap conditions and the worst flow induced vibration effects that might take place over the fuel life time. The fuel rod perforation times calculated from the out-of-pile tests are greater than 1933 days for 2 μm oxidized fuel rods with a 100 μm grid-to-rod gap, whereas those estimated from in-reactor fretting wear failure database may be about in the range of between 60 and 100 days. This large discrepancy in fuel rod perforation may occur due to irradiation-induced cladding oxide microstructure changes on the one hand and a temperature gradient-induced hydrogen content profile across the cladding metal region on the other hand, which may accelerate brittleness in the grid-contacting cladding oxide and metal regions during the reactor operation. A three-phase grid-to-rod fretting wear model is proposed to simulate in-reactor fretting wear progress into the cladding, considering the microstructure changes of the cladding oxide and the hydrogen content profile across the cladding metal region combined with the temperature gradient. The out-of-pile tests cannot be directly applicable to the prediction of in-reactor fretting wear-induced cladding perforations but they can be used only for evaluating a relative wear resistance of one grid design against the other grid design.

Mechanisms of fretting-fatigue of titanium alloys

Energy Technology Data Exchange (ETDEWEB)

Antoniou, R A; Radtke, T C [Defence Sci. and Technol. Organ., Melbourne, Vic. (Australia). Aeronautical and Maritime Res. Lab.

1997-09-30

The effect of continuous fretting in air at 20 C on fatigue performance has been studied for Ti-17 and Ti-6Al-4V, high strength titanium alloys used for gas-turbine fan and compressor disks and blades, respectively. The effect of fretting was to reduce the fatigue stress limit from 700 MPa for plain fatigue to 200 MPa for fretting-fatigue. A number of models, supported by metallographic and fractographic evidence, are proposed which explain (i) how the cyclic loading of individual asperities results in crack initiation; (ii) the formation of multiple cracks; (iii) the existence of non-propagating cracks; and (iv) how fretting influences crack propagation once fatigue cracks have formed. (orig.) 46 refs.

Direct assessment of the effect of the Gly380Arg achondroplasia mutation on FGFR3 dimerization using quantitative imaging FRET.

Directory of Open Access Journals (Sweden)

Jesse Placone

Full Text Available The Gly380Arg mutation in FGFR3 is the genetic cause for achondroplasia (ACH, the most common form of human dwarfism. The mutation has been proposed to increase FGFR3 dimerization, but the dimerization propensities of wild-type and mutant FGFR3 have not been compared. Here we use quantitative imaging FRET to characterize the dimerization of wild-type FGFR3 and the ACH mutant in plasma membrane-derived vesicles from HEK293T cells. We demonstrate a small, but statistically significant increase in FGFR3 dimerization due to the ACH mutation. The data are consistent with the idea that the ACH mutation causes a structural change which affects both the stability and the activity of FGFR3 dimers in the absence of ligand.

Using Three-color Single-molecule FRET to Study the Correlation of Protein Interactions.

Science.gov (United States)

Götz, Markus; Wortmann, Philipp; Schmid, Sonja; Hugel, Thorsten

2018-01-30

Single-molecule Förster resonance energy transfer (smFRET) has become a widely used biophysical technique to study the dynamics of biomolecules. For many molecular machines in a cell proteins have to act together with interaction partners in a functional cycle to fulfill their task. The extension of two-color to multi-color smFRET makes it possible to simultaneously probe more than one interaction or conformational change. This not only adds a new dimension to smFRET experiments but it also offers the unique possibility to directly study the sequence of events and to detect correlated interactions when using an immobilized sample and a total internal reflection fluorescence microscope (TIRFM). Therefore, multi-color smFRET is a versatile tool for studying biomolecular complexes in a quantitative manner and in a previously unachievable detail. Here, we demonstrate how to overcome the special challenges of multi-color smFRET experiments on proteins. We present detailed protocols for obtaining the data and for extracting kinetic information. This includes trace selection criteria, state separation, and the recovery of state trajectories from the noisy data using a 3D ensemble Hidden Markov Model (HMM). Compared to other methods, the kinetic information is not recovered from dwell time histograms but directly from the HMM. The maximum likelihood framework allows us to critically evaluate the kinetic model and to provide meaningful uncertainties for the rates. By applying our method to the heat shock protein 90 (Hsp90), we are able to disentangle the nucleotide binding and the global conformational changes of the protein. This allows us to directly observe the cooperativity between the two nucleotide binding pockets of the Hsp90 dimer.

Investigation of fretting behaviour in pressure armour layers of flexible pipes

Science.gov (United States)

Don Rasika Perera, Solangarachchige

The incidence of fretting damage in the pressure armour wires of flexible pipes used in offshore oil explorations has been investigated. A novel experimental facility which is capable of simulating nub and valley contact conditions of interlocking wire winding with dynamic slip, representative of actual pipe loading, has been developed. The test set-up is equipped with a state of the art data acquisition system and a controller with transducers to measure and control the normal load, slip amplitude and friction force at the contact, in addition to the hoop stress in the wire. Tests were performed with selected loading and the fretted regions were examined using optical microscopy techniques. Results show that the magnitude of contact loading and the slip amplitude have a distinct influence on surface damage. Surface cracks originated from a fretting scar were observed at high contact loads in mixed slip sliding while surface damage predominantly due to wear was observed under gross slip. The position of surface cracks and the wear profile have been related to the contact pressure distribution. The evolution of friction force and surface damage under different slip and normal pressure conditions has been analysed. A fracture mechanics based numerical procedure has been developed to analyse the fretting damage behaviour. A severity parameter is proposed in order to ascertain whether the crack growth is in mode I or mode II cracking. The analysis show the influence of mode II cracking in the early stages of crack growth following which the crack deviates in the mode I direction making mode I the dominant crack propagation mechanism. The crack path determined by the numerical procedure correlates well with the experimental results. A numerical analysis was carried out for the fretting fatigue condition where a cyclic bulk stress superimposes with the friction force. The analysis correlates well with short crack growth behaviour. The analysis confirms that fretting is a

Impact Fretting Wear Behavior of Alloy 690 Tubes in Dry and Deionized Water Conditions

Institute of Scientific and Technical Information of China (English)

Zhen-Bing Cai; Jin-Fang Peng; Hao Qian; Li-Chen Tang; Min-Hao Zhu

2017-01-01

The impact fretting wear has largely occurred at nuclear power device induced by the flow-induced vibration,and it will take potential hazards to the service of the equipment.However,the present study focuses on the tangential fretting wear of alloy 690 tubes.Research on impact fretting wear of alloy 690 tubes is limited and the related research is imminent.Therefore,impact fretting wear behavior of alloy 690 tubes against 304 stainless steels is investigated.Deionized water is used to simulate the flow environment of the equipment,and the dry environment is used for comparison.Varied analytical techniques are employed to characterize the wear and tribochemical behavior during impact fretting wear.Characterization results indicate that cracks occur at high impact load in both water and dry equipment;however,the water as a medium can significantly delay the cracking time.The crack propagation behavior shows a jagged shape in the water,but crack extended disorderly in dry equipment because the water changed the stress distribution and retarded the friction heat during the wear process.The SEM and XPS analysis shows that the main failure mechanisms of the tube under impact fretting are fatigue wear and friction oxidation.The effect of medium(water) on fretting wear is revealed,which plays a potential and promising role in the service of nuclear power device and other flow equipments.

Design improvement for fretting-wear reduction of HANARO fuel assembly

Energy Technology Data Exchange (ETDEWEB)

Cho, Yeong Garp; Chae, H. T.; Ryu, J. S.; Kim, H. R

2000-06-01

In the course of the visual inspection of the fuel assemblies un-loaded from the reactor core in December 1996, it was observed that many of fuel assemblies had mechanical damages on some components. The major damage was the freting-wear on spacer plates and endplates due to the flow induced vibration of the fuel assembly in the flow tube. Since the reactor is activated and the system modification for complete removal of the driving factors of the vibration of fuel assemblies is practically very difficult, the focus has been on the design change of the fuel assemblies. Consequently, various design changes were proposed to strengthen the wear resistance of the components based on the evaluation of the visual inspection results. The validity of the proposals was verified through the performance tests for the modified components, and the vibration test and endurance test for the fuel assemblies using the single-channel test rig(SCTR) in AECL.The subsequent design changes were additionally proposed based on the visual inspections for the fuel assemblies that had been fabricated according to the first design change and loaded in the core. As the effects of the first design change, the fretting-wear of spacer plates was remarkably reduced and the period until fretting-wear damage was extended by 60% for the first modified 36-rod fuel assembly. It is too early to say the endurance life time for the first modified 18-rod fuel assembly because of insufficient statistical data of only two bundles damaged, but the fretting-wear at the bottom endplate slot was reduced to about 50%. The second modified fuel assemblies, that were not loaded into the core yet, are expected to meet the design requirements for the core residence time due to strengthening the weak parts from the fretting-wear point of view. This report describes design changes and tests for fuel assemblies of HANARO to reduce the fretting-wear, and estimates the effects of design improvement quantitatively compared

Design improvement for fretting-wear reduction of HANARO fuel assembly

International Nuclear Information System (INIS)

Cho, Yeong Garp; Chae, H. T.; Ryu, J. S.; Kim, H. R.

2000-06-01

In the course of the visual inspection of the fuel assemblies un-loaded from the reactor core in December 1996, it was observed that many of fuel assemblies had mechanical damages on some components. The major damage was the freting-wear on spacer plates and endplates due to the flow induced vibration of the fuel assembly in the flow tube. Since the reactor is activated and the system modification for complete removal of the driving factors of the vibration of fuel assemblies is practically very difficult, the focus has been on the design change of the fuel assemblies. Consequently, various design changes were proposed to strengthen the wear resistance of the components based on the evaluation of the visual inspection results. The validity of the proposals was verified through the performance tests for the modified components, and the vibration test and endurance test for the fuel assemblies using the single-channel test rig(SCTR) in AECL.The subsequent design changes were additionally proposed based on the visual inspections for the fuel assemblies that had been fabricated according to the first design change and loaded in the core. As the effects of the first design change, the fretting-wear of spacer plates was remarkably reduced and the period until fretting-wear damage was extended by 60% for the first modified 36-rod fuel assembly. It is too early to say the endurance life time for the first modified 18-rod fuel assembly because of insufficient statistical data of only two bundles damaged, but the fretting-wear at the bottom endplate slot was reduced to about 50%. The second modified fuel assemblies, that were not loaded into the core yet, are expected to meet the design requirements for the core residence time due to strengthening the weak parts from the fretting-wear point of view. This report describes design changes and tests for fuel assemblies of HANARO to reduce the fretting-wear, and estimates the effects of design improvement quantitatively compared

QD-Based FRET Probes at a Glance

Directory of Open Access Journals (Sweden)

Armen Shamirian

2015-06-01

Full Text Available The unique optoelectronic properties of quantum dots (QDs give them significant advantages over traditional organic dyes, not only as fluorescent labels for bioimaging, but also as emissive sensing probes. QD sensors that function via manipulation of fluorescent resonance energy transfer (FRET are of special interest due to the multiple response mechanisms that may be utilized, which in turn imparts enhanced flexibility in their design. They may also function as ratiometric, or “color-changing” probes. In this review, we describe the fundamentals of FRET and provide examples of QD-FRET sensors as grouped by their response mechanisms such as link cleavage and structural rearrangement. An overview of early works, recent advances, and various models of QD-FRET sensors for the measurement of pH and oxygen, as well as the presence of metal ions and proteins such as enzymes, are also provided.

Calculated and experimental research of WWER-1000 assembly vibration and fretting damage

International Nuclear Information System (INIS)

Drozdov, Y.; Afanasyev, A.; Makarov, V.; Tutnov, A.; Tutnov, A.; Alekseev, E.

2008-01-01

The report covers the methods and results of the latest analytical and experimental studies of fretting corrosion and natural vibrations of a WWER-1000 reactor fuel assemblies (FA). The process of fretting-corrosion was investigated using a multi-specimen facility that simulated fragments of fuel rod-to-spacer grid and lower support grid mating units. A computational model was developed for vibrations in the mechanical system of a fuel rod fragment and a spacer grid fragment. A calculational and experimental modal analysis of a FA was performed. Natural frequencies, modes and decrements of FA vibrations were determined and a satisfactory coincidence of analytical and experimental results was obtained. The assessment of fretting-corrosion process dynamics was made and its dependences on operational factors were obtained. (authors)

A method to quantify FRET stoichiometry with phasor plot analysis and acceptor lifetime ingrowth.

Science.gov (United States)

Chen, WeiYue; Avezov, Edward; Schlachter, Simon C; Gielen, Fabrice; Laine, Romain F; Harding, Heather P; Hollfelder, Florian; Ron, David; Kaminski, Clemens F

2015-03-10

FRET is widely used for the study of protein-protein interactions in biological samples. However, it is difficult to quantify both the FRET efficiency (E) and the affinity (Kd) of the molecular interaction from intermolecular FRET signals in samples of unknown stoichiometry. Here, we present a method for the simultaneous quantification of the complete set of interaction parameters, including fractions of bound donors and acceptors, local protein concentrations, and dissociation constants, in each image pixel. The method makes use of fluorescence lifetime information from both donor and acceptor molecules and takes advantage of the linear properties of the phasor plot approach. We demonstrate the capability of our method in vitro in a microfluidic device and also in cells, via the determination of the binding affinity between tagged versions of glutathione and glutathione S-transferase, and via the determination of competitor concentration. The potential of the method is explored with simulations. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Probing protein-lipid interactions by FRET between membrane fluorophores

Science.gov (United States)

Trusova, Valeriya M.; Gorbenko, Galyna P.; Deligeorgiev, Todor; Gadjev, Nikolai

2016-09-01

Förster resonance energy transfer (FRET) is a powerful fluorescence technique that has found numerous applications in medicine and biology. One area where FRET proved to be especially informative involves the intermolecular interactions in biological membranes. The present study was focused on developing and verifying a Monte-Carlo approach to analyzing the results of FRET between the membrane-bound fluorophores. This approach was employed to quantify FRET from benzanthrone dye ABM to squaraine dye SQ-1 in the model protein-lipid system containing a polycationic globular protein lysozyme and negatively charged lipid vesicles composed of phosphatidylcholine and phosphatidylglycerol. It was found that acceptor redistribution between the lipid bilayer and protein binding sites resulted in the decrease of FRET efficiency. Quantification of this effect in terms of the proposed methodology yielded both structural and binding parameters of lysozyme-lipid complexes.

Nuclear Fuel Fretting Mechanisms in a Room Temperature Unlubricated Condition

Energy Technology Data Exchange (ETDEWEB)

Lee, Young Ho; Kim, Hyung Kyu [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2008-10-15

Recently, efforts for evaluating the fretting wear mechanism have been carried out by many researchers in various conditions. In an unlubricated condition, especially, effects of a wear debris and/or its layer on the fretting wear behavior were proposed that the formation of a well-developed glaze layer has a beneficial effect for decreasing a friction coefficient. Otherwise, a wear rate was accelerated by a third-body abrasion. At this time, it is well known that wear debris behaviors are affected by test variables such as a temperature, environment, material characteristics, etc. In a nuclear fuel fretting, however, its contact condition is quite different when compared with general fretting wear studies and could be summarized as the following; first, a fuel rod is supported by spacer grid springs and dimples that were elastically deformable. This results in a unique friction loop and a different fretting mechanism when a fuel rod is vibrated due to a flow-induced vibration (FIV). Next, it is possible that some region of the wear scar area with a specific spring shape condition could be hidden due to different wear debris behavior. So, some of the wear debris layers could be found on the worn surfaces in previous studies even though fretting wear tests were performed in a water lubricated condition. Finally, initial contact condition could be changed both an actual operating condition in power plants (i.e. high temperature and pressurized water (HTHP) under severe irradiation conditions) and the fretting wear tests for evaluating the wear resistant spring in lab conditions (i.e. from room temperature to HTHP without irradiation conditions) due to material degradations and the formation of the wear scar, respectively. In summary, the spring shape effect and the variation of the contact condition with increasing fretting cycle should be evaluated in order to improve the wear resistance of the spacer grid spring. So, in this study, fretting wear tests have been

Fretting wear damage of HexTOOL{sup TM} composite depending on the different fibre orientations

Energy Technology Data Exchange (ETDEWEB)

Terekhina, S; Salvia, M; Fouvry, S [Laboratoire de Tribologie et Dynamique des Systemes, UMR CNRS ECL ENISE ENSMSE 5513, Ecole Centrale de Lyon, 69134 Ecully cedex (France); Malysheva, G; Tarasova, T, E-mail: svetlana.terekhina@ec-lyon.fr, E-mail: svetlanaterekhina@yandex.ru [Bauman Moscow State Technical University, 105005 Moscow, 5, 2nd Baumanskaya str (Russian Federation)

2009-09-15

The composites have drawn considerable interest in the mould processes. The vibrations and fatigue stresses induced in the moulds made evident to characterize the composite HexTOOL{sup TM} under fretting conditions. Fretting is a small-amplitude oscillatory motion between contacting surfaces. The running conditions fretting maps (RCFM) of composite at ambient conditions were established. The influence of different fiber orientations of HexTOOL{sup TM} composite on the wear kinetics was shown. An energy wear approach was developed. According to results of dynamic mechanical analysis (DMA), the viscoelastic properties of composite material were obtained.

Assessment of Corrosion, Fretting, and Material Loss of Retrieved Modular Total Knee Arthroplasties.

Science.gov (United States)

Martin, Audrey J; Seagers, Kirsten A; Van Citters, Douglas W

2017-07-01

Modular junctions in total hip arthroplasties have been associated with fretting, corrosion, and debris release. The purpose of this study is to analyze damage severity in total knee arthroplasties of a single design by qualitative visual assessment and quantitative material loss measurements to evaluate implant performance and patient impact via material loss. Twenty-two modular knee retrievals of the same manufacturer were identified from an institutional review board-approved database. Junction designs included tapers with an axial screw and tapers with a radial screw. Constructs consisted of 2 metal alloys: CoCr and Ti6Al4V. Components were qualitatively scored and quantitatively measured for corrosion and fretting. Negative values represent adhered material. Statistical differences were analyzed using sign tests. Correlations were tested with a Spearman rank order test (P corrosion than other components, suggesting preferential corrosion when interfacing with Ti6Al4V. Overall, although corrosion was noted in this series, material loss was low, and none were revised for clinical metal-related reaction. This suggests the clinical impact from corrosion in total knee arthroplasty is low. Copyright © 2017 Elsevier Inc. All rights reserved.

Intercalating dye as an acceptor in quantum-dot-mediated FRET

International Nuclear Information System (INIS)

Lim, Teck Chuan; Bailey, Vasudev J; Wang, T-H; Ho, Y-P

2008-01-01

Fluorescence resonance energy transfer (FRET) is a popular tool to study intermolecular distances and characterize structural or conformational changes of biological macromolecules. We investigate a novel inorganic/organic FRET pair with quantum dots (QDs) as donors and DNA intercalating dyes, BOBO-3, as acceptors by using DNA as a linker. Typically, FRET efficiency increases with the number of stained DNA linked to a QD. However, with the use of intercalating dyes, we demonstrate that FRET efficiency at a fixed DNA:QD ratio can be further enhanced by increasing the number of dyes stained to a DNA strand through the use of an increased staining dye/bp ratio. We exploit this flexibility in the staining ratio to maintain a high FRET efficiency of >0.90 despite a sixfold decrease in DNA concentration. Having characterized this new QD-mediated FRET system, we test this system in a cellular environment using nanocomplexes generated by encapsulating DNA with commercial non-viral gene carriers. Using this novel FRET pair, we are able to monitor the configuration changes and fate of the DNA nanocomplexes during intracellular delivery, thereby providing an insight into the mechanistic study of gene delivery

The Leakage determination on corrosion fretting machine

International Nuclear Information System (INIS)

Sriyono; Satmoko, Ari; Hafid, Abdul; Febrianto; Prasetio, Joko; Abtokhi; Sumarno, Edy; Handoyo, Ismu; Hidayati, Nur Rahmah; Histori

1998-01-01

Fretting machine is an experimental loop to learn fretting corrosion phenomena wich is caused by loading and vibration. On the steam generator, one of the corrosion process that's occurred, it can be caused by vibration between tubes and bending material. Because of high flow rate inside the tube, the high frequency vibration will appeared so it can make the corrosion on bending material more faster. This process can be simulate by fretting machine. This machine has already damage because of leakage. So it will be repaired by dismantling, radiography testing and redrawing. from the result of radiography, the leakage is caused by cracking on bellows seals of the dynamic main support

Application of Influence Function Method to the Fretting Wear Problems

Energy Technology Data Exchange (ETDEWEB)

Lee, Choon Yeol; Tian, Li Si; Bae, Joon Woo; Chai, Young Suck [Yeungnam University, Gyongsan (Korea, Republic of)

2006-07-01

Numerical analysis by influence function method (IFM) is demonstrated in this study in order to investigate the fretting wear problems on the secondary side of the steam generator, caused by flow induced vibration. Two-dimensional numerical contact model in terms of Cauchy integral equation is developed. The distributions of normal pressures, shear stresses and displacement fields are derived between two contact bodies which have similar elastic properties. The work rate model is adopted to find the wear amounts between two materials. The results are compared with the solutions by finite element analyses, which show the utilization of the present method to the fretting wear problems.

Application of Influence Function Method to the Fretting Wear Problems

International Nuclear Information System (INIS)

Lee, Choon Yeol; Tian, Li Si; Bae, Joon Woo; Chai, Young Suck

2006-01-01

Numerical analysis by influence function method (IFM) is demonstrated in this study in order to investigate the fretting wear problems on the secondary side of the steam generator, caused by flow induced vibration. Two-dimensional numerical contact model in terms of Cauchy integral equation is developed. The distributions of normal pressures, shear stresses and displacement fields are derived between two contact bodies which have similar elastic properties. The work rate model is adopted to find the wear amounts between two materials. The results are compared with the solutions by finite element analyses, which show the utilization of the present method to the fretting wear problems

Reconstruction of calmodulin single-molecule FRET states, dye interactions, and CaMKII peptide binding by MultiNest and classic maximum entropy

Science.gov (United States)

DeVore, Matthew S.; Gull, Stephen F.; Johnson, Carey K.

2013-08-01

We analyzed single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest compares favorably with maximum entropy analysis for simulated data, judged by the Bayesian evidence. FRET efficiency distributions recovered for calmodulin labeled with two different FRET dye pairs depended on the dye pair and changed upon Ca2+ binding. We also looked at the FRET efficiency distributions of calmodulin bound to the calcium/calmodulin dependent protein kinase II (CaMKII) binding domain. For both dye pairs, the FRET efficiency distribution collapsed to a single peak in the case of calmodulin bound to the CaMKII peptide. These measurements strongly suggest that consideration of dye-protein interactions is crucial in forming an accurate picture of protein conformations from FRET data.

Reconstruction of Calmodulin Single-Molecule FRET States, Dye-Interactions, and CaMKII Peptide Binding by MultiNest and Classic Maximum Entropy.

Science.gov (United States)

Devore, Matthew S; Gull, Stephen F; Johnson, Carey K

2013-08-30

We analyze single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest compares favorably with maximum entropy analysis for simulated data, judged by the Bayesian evidence. FRET efficiency distributions recovered for calmodulin labeled with two different FRET dye pairs depended on the dye pair and changed upon Ca 2+ binding. We also looked at the FRET efficiency distributions of calmodulin bound to the calcium/calmodulin dependent protein kinase II (CaMKII) binding domain. For both dye pairs, the FRET efficiency distribution collapsed to a single peak in the case of calmodulin bound to the CaMKII peptide. These measurements strongly suggest that consideration of dye-protein interactions is crucial in forming an accurate picture of protein conformations from FRET data.

Multi step FRET among three laser dyes Pyrene, Acriflavine and Rhodamine B

International Nuclear Information System (INIS)

Saha, Jaba; Dey, Dibyendu; Roy, Arpan Datta; Bhattacharjee, D.; Hussain, Syed Arshad

2016-01-01

Fluorescence Resonance Energy Transfer (FRET) system using three dyes has been demonstrated. It has been observed that multi step energy transfer occurred from Pyrene to Rhodamine B via Acriflavine. Here Acriflavine acts as an antenna to receive energy from Pyrene and transfer the same to Rhodamine B. This multi step FRET system is advantageous compared to the conventional FRET as this can be used to study molecular level interaction beyond conventional FRET distance (1–10 nm) as well as studying multi-branched macromolecules. The introduction of clay enhances the FRET efficiencies among the dye pair, which is an advantage to make the multi step system more useful. Similar approach can be used for increasing FRET efficiencies by using other dyes. - Highlights: • Multi-step FRET occurred from Pyrene (Py) to Rhodamine B (RhB) via Acriflavine (Acf). • Acf acts as an antenna to receive energy from Py and to transfer energy to RhB. • Multi-step FRET can be used to study molecular level interaction beyond 1–10 nm. • Incorporation of nanoclay laponite enhances the energy transfer efficiency.

Derivation of Elastic Stress Concentration Factor Equations for Debris Fretting Flaws in Pressure Tubes of Pressurized Heavy Water Reactors

International Nuclear Information System (INIS)

Kim, Jong Sung; Oh, Young Jin

2014-01-01

If volumetric flaws such as bearing pad fretting flaws and debris fretting flaws are detected in the pressure tubes of pressurized heavy water reactors during in-service inspection, the initiation of fatigue cracks and delayed hydrogen cracking from the detected volumetric flaws shall be assessed by using elastic stress concentration factors in accordance with CSA N285.8-05. The CSA N285.8-05 presents only an approximate formula based on linear elastic fracture mechanics for the debris fretting flaw. In this study, an engineering formula considering the geometric characteristics of the debris fretting flaw in detail was derived using two-dimensional finite element analysis and Kinectrics, Inc.'s engineering procedure with slight modifications. Comparing the application results obtained using the derived formula with the three-dimensional finite element analysis results, it is found that the results obtained using the derived formula agree well with the results of the finite element analysis

a Study on the Fretting Fatigue Life of Zircaloy Alloys

Science.gov (United States)

Kwon, Jae-Do; Park, Dae-Kyu; Woo, Seung-Wan; Chai, Young-Suck

Studies on the strength and fatigue life of machines and structures have been conducted in accordance with the development of modern industries. In particular, fine and repetitive cyclic damage occurring in contact regions has been known to have an impact on fretting fatigue fractures. The main component of zircaloy alloy is Zr, and it possesses good mechanical characteristics at high temperatures. This alloy is used in the fuel rod material of nuclear power plants because of its excellent resistance. In this paper, the effect of the fretting damage on the fatigue behavior of the zircaloy alloy is studied. Further, various types of mechanical tests such as tension and plain fatigue tests are performed. Fretting fatigue tests are performed with a flat-flat contact configuration using a bridge-type contact pad and plate-type specimen. Through these experiments, it is found that the fretting fatigue strength decreases by about 80% as compared to the plain fatigue strength. Oblique cracks are observed in the initial stage of the fretting fatigue, in which damaged areas are found. These results can be used as the basic data for the structural integrity evaluation of corrosion-resisting alloys considering the fretting damages.

Understanding and modeling Förster-type resonance energy transfer (FRET)

CERN Document Server

Hernández Martínez, Pedro Ludwig; Demir, Hilmi Volkan

2017-01-01

This Brief presents a complete study of the generalized theory of Förster-type energy transfer in nanostructures with mixed dimensionality. Here the aim is to obtain a generalized theory of FRET including a comprehensive set of analytical equations for all combinations and configurations of nanostructures and deriving generic expressions for the dimensionality involved. In this brief, the modification of FRET mechanism with respect to the nanostructure serving as the donor vs. the acceptor will be included, focusing on the rate’s distance dependency and the role of the effective dielectric function in FRET, which will be a unique, useful source for those who study and model FRET.

CONSIDERATIONS REGARDING THE FRETTING PHENOMENON USING LEAF SPRINGS

Directory of Open Access Journals (Sweden)

Stefan GHIMIȘI

2015-05-01

Full Text Available The fretting phenomenon represents particulary and complex form of wear who is; generaly, and/or weary of fretting who is produced on the load contact in a relative oscialatory movement lay small amplitude.A simultaneoustly applied tangential force and normal into contact appears a adhesion force

Multispot single-molecule FRET: High-throughput analysis of freely diffusing molecules.

Directory of Open Access Journals (Sweden)

Antonino Ingargiola

Full Text Available We describe an 8-spot confocal setup for high-throughput smFRET assays and illustrate its performance with two characteristic experiments. First, measurements on a series of freely diffusing doubly-labeled dsDNA samples allow us to demonstrate that data acquired in multiple spots in parallel can be properly corrected and result in measured sample characteristics consistent with those obtained with a standard single-spot setup. We then take advantage of the higher throughput provided by parallel acquisition to address an outstanding question about the kinetics of the initial steps of bacterial RNA transcription. Our real-time kinetic analysis of promoter escape by bacterial RNA polymerase confirms results obtained by a more indirect route, shedding additional light on the initial steps of transcription. Finally, we discuss the advantages of our multispot setup, while pointing potential limitations of the current single laser excitation design, as well as analysis challenges and their solutions.

A FRET-based approach for quantitative evaluation of forskolin-induced pendrin trafficking at the plasma membrane in bronchial NCI H292 cells.

Science.gov (United States)

Tamma, Grazia; Ranieri, Marianna; Dossena, Silvia; Di Mise, Annarita; Nofziger, Charity; Svelto, Maria; Paulmichl, Markus; Valenti, Giovanna

2013-01-01

Human pendrin (SLC26A4, PDS) is an integral membrane protein acting as an electroneutral anion exchanger. Loss of function mutations in pendrin protein cause Pendred syndrome, a disorder characterized by sensorineural deafness and a partial iodide organification defect that may lead to thyroid goiter. Additionally, pendrin up-regulation could play a role in the pathogenesis of several diseases including bronchial asthma and chronic obstructive pulmonary disease (COPD). Therefore, monitoring the plasma membrane abundance and trafficking of pendrin in the context of a living cell is crucially important. Trafficking of pendrin to the plasma membrane was monitored by fluorescence resonance energy transfer (FRET), a physical phenomenon occurring between two fluorophores (the FRET donor and acceptor) located in close spatial proximity. Because the efficiency of the energy transfer is inversely proportional to the sixth power of the distance between donor and acceptor, FRET is extremely sensitive to small changes in distance between the donor and acceptor and is therefore a powerful tool to determine protein-protein interactions. FRET studies revealed that forskolin-induced cAMP production is associated with a significant increase of pendrin expression at plasma membrane, which is paralleled by a decrease in intracellular pH. Pendrin transposition to the membrane is accompanied with a partial depolymerization of actin cytoskeleton via Rho-GTPase inhibition. Trafficking to the plasma membrane is critical in the regulation of pendrin activity. Therefore, reliable tools for monitoring and quantifying this phenomenon are highly desirable. © 2014 S. Karger AG, Basel.

Prediction of pressure tube fretting-wear damage due to fuel vibration

International Nuclear Information System (INIS)

Yetisir, M.; Fisher, N.J.

1997-01-01

Fretting marks between fuel bundle bearing pads and pressure tubes have been observed at the inlet end of some Darlington Nuclear Generating Station (NGS) and Bruce NGS fuel channels. The excitation mechanisms that lead to fretting are not fully understood. In this paper, the possibility of bearing pad-to-pressure tube fretting due to turbulence-induced motion of the fuel element is investigated. Numerical simulations indicate that this mechanism by itself is not likely to cause the level of fretting experienced in Darlington and Bruce NGSs. (orig.)

Characterizing the interactions between prolyl isomerase pin1 and phosphatase inhibitor-2 in living cells with FRET and FCS

Science.gov (United States)

Sun, Yuansheng; Wang, Lifu; Jyothikumar, Vinod; Brautigan, David L.; Periasamy, Ammasi

2012-03-01

Phosphatase inhibitor-2 (I2) was discovered as a regulator of protein Ser/Thr phosphatase-1 and is conserved from yeast to human. Binding between purified recombinant I2 from different species and the prolyl isomerase Pin1 has been demonstrated with pull-down assays, size exclusion chromatography and nuclear magnetic resonance spectroscopy. Despite this, questions persist as to whether these proteins associate together in living cells. In this study, we prepared fluorescent protein (FP) fusions of I2 and Pin1 and employed both Förster Resonance Energy Transfer (FRET) and Fluorescence Correlation Spectroscopy (FCS) imaging techniques to characterize their interactions in living cells. In both intensity-based and time-resolved FRET studies, we observed FRET uniformly across whole cells co-expressing I2-Cerulean and Pin1-Venus that was significantly higher than in negative controls expressing Cerulean FP (without fusing to I2) as the FRET donor and Pin1-Venus, showing a specific interaction between I2-Cerulean and Pin1-Venus in living cells. We also observed the co-diffusion of I2-Cerulean and Pin1-mCherry in Fluorescence Cross Correlation Spectroscopy (FCCS) measurements. We further showed that I2 itself as well as I2-Pin1 formed complexes in living cells (predicted from in vitro studies) via a quantitative FRET assay, and demonstrated from FCS measurements that both I2 and Pin1 (fused to Cerulean) are highly mobile in living cells.

On the geometry of the fuel rod supports concerning a fretting wear failure

International Nuclear Information System (INIS)

Kim, Hyung-Kyu; Lee, Young-Ho; Lee, Kang-Hee

2008-01-01

Geometrical conditions of spacer grid springs and dimples of a light water reactor fuel assembly are studied in this paper concerning a fuel rod's fretting wear failure. In this framework, the springs/dimples are categorized from the aspects of their orientation with respect to the fuel axis and the contact types. Possible motions on the contacts between the springs/dimples and fuel rods are estimated by conducting a flow-induced vibration test. Features of the wear scar and depth are investigated by independent fretting wear tests carried out with spring and dimple specimens of typical contact geometries. It is also attempted here to apply the contact mechanics theory to a fuel fretting wear analysis such as the prediction of a wear depth profile and its rate, which is influenced by the contact shape of the springs/dimples. It is shown that the theory can be applied to a dimensional control of a coining for the springs/dimples, which is usually carried out in a thin plate fabrication. From the results, the necessary conditions for a spring/dimple geometry for restraining a fretting wear failure are discussed

Steam generator fretting-wear damage: A summary of recent findings

International Nuclear Information System (INIS)

Guerout, F.M.; Fisher, N.J.

1999-01-01

Flow-induced vibration of steam generator (SG) tubes may sometimes result in fretting-wear damage at the tube-to-support locations. Fretting-wear damage predictions are largely based on experimental data obtained at representative test conditions. Fretting-wear of SG materials has been studied at the Chalk River Laboratories for two decades. Tests are conducted in fretting-wear test machines that simulate SG environmental conditions and tube-to-support dynamic interactions. A new high-temperature force and displacement measuring system was developed to monitor tube-to-support interaction (i.e., work-rate) at operating conditions. This improvement in experimental fretting-wear technology was used to perform a comprehensive study of the effect of various environment and design parameters on SG tube wear damage. This paper summarizes the results of tests performed over the past 4 yr to study the effect of temperature, water chemistry, support geometry, and tube material on fretting-wear. The results show a significant effect of temperature on tube wear damage. Therefore, fretting-wear tests must be performed at operating temperatures in order to be relevant. No significant effect of the type of water treatment on tube wear damage was observed. For predominantly impacting motion, the wear of SG tubes in contact with 410 stainless steel is similar regardless of whether Alloy 690 or Alloy 800 is used as tubing material or whether lattice bars or broached hole supports are used. Based on results presented in this paper, an average wear coefficient value is recommended that is used for the prediction of SG tube wear depth versus time

A FRET-Based Approach for Quantitative Evaluation of Forskolin-Induced Pendrin Trafficking at the Plasma Membrane in Bronchial NCI H292 Cells

Directory of Open Access Journals (Sweden)

Grazia Tamma

2013-12-01

Full Text Available Background: Human pendrin (SLC26A4, PDS is an integral membrane protein acting as an electroneutral anion exchanger. Loss of function mutations in pendrin protein cause Pendred syndrome, a disorder characterized by sensorineural deafness and a partial iodide organification defect that may lead to thyroid goiter. Additionally, pendrin up-regulation could play a role in the pathogenesis of several diseases including bronchial asthma and chronic obstructive pulmonary disease (COPD. Therefore, monitoring the plasma membrane abundance and trafficking of pendrin in the context of a living cell is crucially important. Methods: Trafficking of pendrin to the plasma membrane was monitored by fluorescence resonance energy transfer (FRET, a physical phenomenon occurring between two fluorophores (the FRET donor and acceptor located in close spatial proximity. Because the efficiency of the energy transfer is inversely proportional to the sixth power of the distance between donor and acceptor, FRET is extremely sensitive to small changes in distance between the donor and acceptor and is therefore a powerful tool to determine protein-protein interactions. Results: FRET studies revealed that forskolin-induced cAMP production is associated with a significant increase of pendrin expression at plasma membrane, which is paralleled by a decrease in intracellular pH. Pendrin transposition to the membrane is accompanied with a partial depolymerization of actin cytoskeleton via Rho-GTPase inhibition. Conclusion: Trafficking to the plasma membrane is critical in the regulation of pendrin activity. Therefore, reliable tools for monitoring and quantifying this phenomenon are highly desirable.

Prediction of pressure tube fretting-wear damage due to fuel vibration

Energy Technology Data Exchange (ETDEWEB)

Yetisir, M; Fisher, N J [Atomic Energy of Canada Ltd., Chalk River, ON (Canada)

1996-12-31

Fretting marks between fuel bundle bearing pads and pressure tubes have been observed at the inlet end of some Darlington NGS (nuclear generating station) and Bruce NGS fuel channels. The excitation mechanisms that lead to fretting are not fully understood. In this paper, the possibility of bearing pad-to-pressure tube fretting due to turbulence-induced motion of the fuel element is investigated. Numerical simulations indicate that this mechanism by itself is not likely to cause the level of fretting experienced in Darlington and Bruce NGS`s (nuclear generating stations). (author). 12 refs., 2 tabs., 11 figs.

Two-dimensional Forster resonance energy transfer (2-D FRET) and the membrane raft hypothesis

OpenAIRE

Acasandrei, Maria; Dale, Robert; VAN DE VEN, Martin; AMELOOT, Marcel

2006-01-01

A model for analyzing Forster resonance energy transfer (FRET) data in relation to the cell plasma membrane raft hypothesis is developed to take into account: (a) the distribution of FRET donors and acceptors at the surface of probing antibody fragments specific for a putative raft component; (b) partitioning of the raft component between raft and non-raft areas of the membrane; and (c) the dependence of the raft partition on the expression level of the considered component. Analysis of relev...

A Study on Fretting Behavior in Room Temperature for Inconel Alloy 690

Science.gov (United States)

Kwon, Jae Do; Chai, Young Suck; Bae, Yong Tak; Choi, Sung Jong

The initial crack under fretting condition occurs at lower stress amplitude and lower cycles of cyclic loading than that under plain fatigue condition. The fretting damage, for example, can be observed in fossil and nuclear power plant, aircraft, automobile and petroleum chemical plants etc. INCONEL alloy 690 is a high-chromium nickel alloy having excellent resistance to many corrosive aqueous media and high-temperature atmospheres. This alloy is used extensively in the industries of nuclear power, chemicals, heat-treatment and electronics. In this paper, the effect of fretting damage on fatigue behavior for INCONEL alloy 690 was studied. Also, various kinds of tests on mechanical properties such as hardness, tension and plain fatigue tests are performed. Fretting fatigue tests were carried out with flat-flat contact configuration using a bridge type contact pad and plate type specimen. Through these experiments, it is found that the fretting fatigue strength decreased about 43% compared to the plain fatigue strength. In fretting fatigue, the wear debris is observed on the contact surface, and the oblique micro-cracks are initiated at an earlier stage. These results can be used as the basic data in a structural integrity evaluation of heat and corrosion resistant alloy considering fretting damages.

Fuel bundle to pressure tube fretting in Bruce and Darlington

Energy Technology Data Exchange (ETDEWEB)

Norsworthy, A G; Ditschun, A [Atomic Energy of Canada Ltd., Mississauga, ON (Canada)

1996-12-31

As the fuel channel elongates due to creep, the fuel string moves relative to the inlet until the fuel pads at the inboard end eventually separate from the spacer sleeve, and the fuel resides on the burnish mark of the pressure tube. The bundle is then supported in a fashion which contributes to increased levels of vibration. Those pads which (due to geometric variation) have contact loads with the pressure tube within a certain range, vibrate, and cause significant fretting on the burnish mark, and further along at the midplane of the bundle. Inspection of the pressure tubes in Bruce A, Bruce B, and Darlington has revealed fret damage up to 0.55 mm at the burnish mark and slightly lower than this at the inlet bundle midplane. To date, all fret marks have been dealt with successfully without the need for tube replacement, but a program of work has been initiated to understand the mechanism and reduce the fretting. Such understanding is necessary to guide future design changes to the fuel bundle, to guide future inspection programs, to guide maintenance programs, and for longer term strategic planning. This paper discusses how the understanding of fretting has evolved and outlines a current hypothesis for the mechanism of fretting. The role of bundle geometry, excitation forces, and reactor conditions are reviewed, along with options under consideration to mitigate damage. (author). 4 refs., 2 tabs., 13 figs.

Fuel bundle to pressure tube fretting in Bruce and Darlington

International Nuclear Information System (INIS)

Norsworthy, A.G.; Ditschun, A.

1995-01-01

As the fuel channel elongates due to creep, the fuel string moves relative to the inlet until the fuel pads at the inboard end eventually separate from the spacer sleeve, and the fuel resides on the burnish mark of the pressure tube. The bundle is then supported in a fashion which contributes to increased levels of vibration. Those pads which (due to geometric variation) have contact loads with the pressure tube within a certain range, vibrate, and cause significant fretting on the burnish mark, and further along at the midplane of the bundle. Inspection of the pressure tubes in Bruce A, Bruce B, and Darlington has revealed fret damage up to 0.55 mm at the burnish mark and slightly lower than this at the inlet bundle midplane. To date, all fret marks have been dealt with successfully without the need for tube replacement, but a program of work has been initiated to understand the mechanism and reduce the fretting. Such understanding is necessary to guide future design changes to the fuel bundle, to guide future inspection programs, to guide maintenance programs, and for longer term strategic planning. This paper discusses how the understanding of fretting has evolved and outlines a current hypothesis for the mechanism of fretting. The role of bundle geometry, excitation forces, and reactor conditions are reviewed, along with options under consideration to mitigate damage. (author). 4 refs., 2 tabs., 13 figs

Standard test method for damage to contacting solid surfaces under fretting conditions

CERN Document Server

American Society for Testing and Materials. Philadelphia

2010-01-01

1.1 This test method covers the studying or ranking the susceptibility of candidate materials to fretting corrosion or fretting wear for the purposes of material selection for applications where fretting corrosion or fretting wear can limit serviceability. 1.2 This test method uses a tribological bench test apparatus with a mechanism or device that will produce the necessary relative motion between a contacting hemispherical rider and a flat counterface. The rider is pressed against the flat counterface with a loading mass. The test method is intended for use in room temperature air, but future editions could include fretting in the presence of lubricants or other environments. 1.3 The purpose of this test method is to rub two solid surfaces together under controlled fretting conditions and to quantify the damage to both surfaces in units of volume loss for the test method. 1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.5...

Classic maximum entropy recovery of the average joint distribution of apparent FRET efficiency and fluorescence photons for single-molecule burst measurements.

Science.gov (United States)

DeVore, Matthew S; Gull, Stephen F; Johnson, Carey K

2012-04-05

We describe a method for analysis of single-molecule Förster resonance energy transfer (FRET) burst measurements using classic maximum entropy. Classic maximum entropy determines the Bayesian inference for the joint probability describing the total fluorescence photons and the apparent FRET efficiency. The method was tested with simulated data and then with DNA labeled with fluorescent dyes. The most probable joint distribution can be marginalized to obtain both the overall distribution of fluorescence photons and the apparent FRET efficiency distribution. This method proves to be ideal for determining the distance distribution of FRET-labeled biomolecules, and it successfully predicts the shape of the recovered distributions.

A New Generation of FRET Sensors for Robust Measurement of Gαi1, Gαi2 and Gαi3 Activation Kinetics in Single Cells.

Directory of Open Access Journals (Sweden)

Jakobus van Unen

Full Text Available G-protein coupled receptors (GPCRs can activate a heterotrimeric G-protein complex with subsecond kinetics. Genetically encoded biosensors based on Förster resonance energy transfer (FRET are ideally suited for the study of such fast signaling events in single living cells. Here we report on the construction and characterization of three FRET biosensors for the measurement of Gαi1, Gαi2 and Gαi3 activation. To enable quantitative long-term imaging of FRET biosensors with high dynamic range, fluorescent proteins with enhanced photophysical properties are required. Therefore, we use the currently brightest and most photostable CFP variant, mTurquoise2, as donor fused to Gαi subunit, and cp173Venus fused to the Gγ2 subunit as acceptor. The Gαi FRET biosensors constructs are expressed together with Gβ1 from a single plasmid, providing preferred relative expression levels with reduced variation in mammalian cells. The Gαi FRET sensors showed a robust response to activation of endogenous or over-expressed alpha-2A-adrenergic receptors, which was inhibited by pertussis toxin. Moreover, we observed activation of the Gαi FRET sensor in single cells upon stimulation of several GPCRs, including the LPA2, M3 and BK2 receptor. Furthermore, we show that the sensors are well suited to extract kinetic parameters from fast measurements in the millisecond time range. This new generation of FRET biosensors for Gαi1, Gαi2 and Gαi3 activation will be valuable for live-cell measurements that probe Gαi activation.

Extraction of information on macromolecular interactions from fluorescence micro-spectroscopy measurements in the presence and absence of FRET

Science.gov (United States)

Raicu, Valerică

2018-06-01

Investigations of static or dynamic interactions between proteins or other biological macromolecules in living cells often rely on the use of fluorescent tags with two different colors in conjunction with adequate theoretical descriptions of Förster Resonance Energy Transfer (FRET) and molecular-level micro-spectroscopic technology. One such method based on these general principles is FRET spectrometry, which allows determination of the quaternary structure of biomolecules from cell-level images of the distributions, or spectra of occurrence frequency of FRET efficiencies. Subsequent refinements allowed combining FRET frequency spectra with molecular concentration information, thereby providing the proportion of molecular complexes with various quaternary structures as well as their binding/dissociation energies. In this paper, we build on the mathematical principles underlying FRET spectrometry to propose two new spectrometric methods, which have distinct advantages compared to other methods. One of these methods relies on statistical analysis of color mixing in subpopulations of fluorescently tagged molecules to probe molecular association stoichiometry, while the other exploits the color shift induced by FRET to also derive geometric information in addition to stoichiometry. The appeal of the first method stems from its sheer simplicity, while the strength of the second consists in its ability to provide structural information.

Wave propagation in coated cylinders with reference to fretting fatigue

Indian Academy of Sciences (India)

is to study stress wave propagation in cylinders with reference to high frequency fretting. ... The motivation for studying of fretting fatigue at higher frequency is to investigate the ... Hence focus in this work is given to thin rods and cylinders. The.

Simultaneous live cell imaging using dual FRET sensors with a single excitation light.

Directory of Open Access Journals (Sweden)

Yusuke Niino

Full Text Available Fluorescence resonance energy transfer (FRET between fluorescent proteins is a powerful tool for visualization of signal transduction in living cells, and recently, some strategies for imaging of dual FRET pairs in a single cell have been reported. However, these necessitate alteration of excitation light between two different wavelengths to avoid the spectral overlap, resulting in sequential detection with a lag time. Thus, to follow fast signal dynamics or signal changes in highly motile cells, a single-excitation dual-FRET method should be required. Here we reported this by using four-color imaging with a single excitation light and subsequent linear unmixing to distinguish fluorescent proteins. We constructed new FRET sensors with Sapphire/RFP to combine with CFP/YFP, and accomplished simultaneous imaging of cAMP and cGMP in single cells. We confirmed that signal amplitude of our dual FRET measurement is comparable to of conventional single FRET measurement. Finally, we demonstrated to monitor both intracellular Ca(2+ and cAMP in highly motile cardiac myocytes. To cancel out artifacts caused by the movement of the cell, this method expands the applicability of the combined use of dual FRET sensors for cell samples with high motility.

APPL proteins FRET at the BAR: direct observation of APPL1 and APPL2 BAR domain-mediated interactions on cell membranes using FRET microscopy.

Directory of Open Access Journals (Sweden)

Heidi J Chial

2010-08-01

Full Text Available Human APPL1 and APPL2 are homologous RAB5 effectors whose binding partners include a diverse set of transmembrane receptors, signaling proteins, and phosphoinositides. APPL proteins associate dynamically with endosomal membranes and are proposed to function in endosome-mediated signaling pathways linking the cell surface to the cell nucleus. APPL proteins contain an N-terminal Bin/Amphiphysin/Rvs (BAR domain, a central pleckstrin homology (PH domain, and a C-terminal phosphotyrosine binding (PTB domain. Previous structural and biochemical studies have shown that the APPL BAR domains mediate homotypic and heterotypic APPL-APPL interactions and that the APPL1 BAR domain forms crescent-shaped dimers. Although previous studies have shown that APPL minimal BAR domains associate with curved cell membranes, direct interaction between APPL BAR domains on cell membranes in vivo has not been reported.Herein, we used a laser-scanning confocal microscope equipped with a spectral detector to carry out fluorescence resonance energy transfer (FRET experiments with cyan fluorescent protein/yellow fluorescent protein (CFP/YFP FRET donor/acceptor pairs to examine interactions between APPL minimal BAR domains at the subcellular level. This comprehensive approach enabled us to evaluate FRET levels in a single cell using three methods: sensitized emission, standard acceptor photobleaching, and sequential acceptor photobleaching. We also analyzed emission spectra to address an outstanding controversy regarding the use of CFP donor/YFP acceptor pairs in FRET acceptor photobleaching experiments, based on reports that photobleaching of YFP converts it into a CFP-like species.All three methods consistently showed significant FRET between APPL minimal BAR domain FRET pairs, indicating that they interact directly in a homotypic (i.e., APPL1-APPL1 and APPL2-APPL2 and heterotypic (i.e., APPL1-APPL2 manner on curved cell membranes. Furthermore, the results of our experiments

Turbulence induced Fretting-wear characteristics of steam generator helical tubes

International Nuclear Information System (INIS)

Jhung, Myung Jo; Jo, Jong Chull; Kim, Hho Jung; Yune, Young Gill; Yu, Seon Oh

2005-01-01

This study addresses safety assessment of the potential for fretting-wear damages on steam generator helical tubes due to turbulence-induced vibration in operating nuclear power plants. To get the natural frequency, corresponding mode shape and participation factor, modal analyses are performed for helical type tubes with various conditions. Special emphases are put on the effects of coil diameter and the number of turns on the modal and fretting wear characteristics of tubes. Also, investigated are the effects of external pressure on the tube modal characteristics as well as the effects of turbulence induced vibration on the fretting-wear characteristics of tubes

Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma

Directory of Open Access Journals (Sweden)

James Miles

2017-12-01

General significance: The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment.

Fretting fatigue behaviour of Ni-free high-nitrogen stainless steel in a simulated body fluid

Directory of Open Access Journals (Sweden)

Norio Maruyama, Sachiko Hiromoto, Eiji Akiyama and Morihiko Nakamura

2013-01-01

Full Text Available Fretting fatigue behaviour of Ni-free high-nitrogen steel (HNS with a yield strength of about 800 MPa, which was prepared by nitrogen gas pressurized electroslag remelting, was studied in air and in phosphate-buffered saline (PBS(-. For comparison, fretting fatigue behaviour of cold-rolled SUS316L steel (SUS316L(CR with similar yield strength was examined. The plain fatigue limit of HNS was slightly lower than that of SUS316L(CR although the former had a higher tensile strength than the latter. The fretting fatigue limit of HNS was higher than that of SUS316L(CR both in air and in PBS(-. A decrease in fatigue limit of HNS by fretting was significantly smaller than that of SUS316L(CR in both environments, indicating that HNS has better fretting fatigue resistance than SUS316L(CR. The decrease in fatigue limit by fretting is discussed taking into account the effect of friction stress due to fretting and the additional influences of wear, tribocorrosion and plastic deformation in the fretted area.

Roughness Influence on Initiation of Fretting Fatigue Scar of Ti-6Al-4V Alloy

Science.gov (United States)

Capitanu, L.; Badita, L. L.; Florescu, V.; Tiganesteanu, C.

2018-01-01

This paper reports on the experimental studies undertaken to detect the early stage when appears the fretting wear of the Ti-6Al-4V alloy used for the hip prostheses. Wear is a critical aspect for estimating the fretting fatigue. Studies were performed on samples of special shape, in order to be able to study the influence of in contact surfaces roughness on the durability to fretting. Fretting buffers, with roughnesses Ra of the contact surface of 0.015 and 0.045 μm, and Ti-6Al-4V samples with roughnesses Ra = 0.045 μm, Ra = 0.075 μm and Ra = 0.19 μm, were used. Testing periods of 3 seconds, 1 minute and 5 minutes were selected to capture the moment of the fretting scar appearance, long before these initiate the eventual fretting cracking. Simultaneously with fretting wear of the surface, the friction coefficient was also measured. From the in time evolution determinations of the fretting wear, it resulted that, under the experimental conditions used, the minimum wear occurs at a certain value of the roughness and not at the minimum roughness. Surprisingly, the minimum friction coefficient does not coincide with the minimum fretting wear.

Terbium to Quantum Dot FRET Bioconjugates for Clinical Diagnostics: Influence of Human Plasma on Optical and Assembly Properties

Directory of Open Access Journals (Sweden)

Niko Hildebrandt

2011-10-01

Full Text Available Förster resonance energy transfer (FRET from luminescent terbium complexes (LTC as donors to semiconductor quantum dots (QDs as acceptors allows extraordinary large FRET efficiencies due to the long Förster distances afforded. Moreover, time-gated detection permits an efficient suppression of autofluorescent background leading to sub-picomolar detection limits even within multiplexed detection formats. These characteristics make FRET-systems with LTC and QDs excellent candidates for clinical diagnostics. So far, such proofs of principle for highly sensitive multiplexed biosensing have only been performed under optimized buffer conditions and interactions between real-life clinical media such as human serum or plasma and LTC-QD-FRET-systems have not yet been taken into account. Here we present an extensive spectroscopic analysis of absorption, excitation and emission spectra along with the luminescence decay times of both the single components as well as the assembled FRET-systems in TRIS-buffer, TRIS-buffer with 2% bovine serum albumin, and fresh human plasma. Moreover, we evaluated homogeneous LTC-QD FRET assays in QD conjugates assembled with either the well-known, specific biotin-streptavidin biological interaction or, alternatively, the metal-affinity coordination of histidine to zinc. In the case of conjugates assembled with biotin-streptavidin no significant interference with the optical and binding properties occurs whereas the histidine-zinc system appears to be affected by human plasma.

Mining the Sinorhizobium meliloti transportome to develop FRET biosensors for sugars, dicarboxylates and cyclic polyols.

Directory of Open Access Journals (Sweden)

Alexandre Bourdès

Full Text Available Förster resonance energy transfer (FRET biosensors are powerful tools to detect biologically important ligands in real time. Currently FRET bisosensors are available for twenty-two compounds distributed in eight classes of chemicals (two pentoses, two hexoses, two disaccharides, four amino acids, one nucleobase, two nucleotides, six ions and three phytoestrogens. To expand the number of available FRET biosensors we used the induction profile of the Sinorhizobium meliloti transportome to systematically screen for new FRET biosensors.Two new vectors were developed for cloning genes for solute-binding proteins (SBPs between those encoding FRET partner fluorescent proteins. In addition to a vector with the widely used cyan and yellow fluorescent protein FRET partners, we developed a vector using orange (mOrange2 and red fluorescent protein (mKate2 FRET partners. From the sixty-nine SBPs tested, seven gave a detectable FRET signal change on binding substrate, resulting in biosensors for D-quinic acid, myo-inositol, L-rhamnose, L-fucose, β-diglucosides (cellobiose and gentiobiose, D-galactose and C4-dicarboxylates (malate, succinate, oxaloacetate and fumarate. To our knowledge, we describe the first two FRET biosensor constructs based on SBPs from Tripartite ATP-independent periplasmic (TRAP transport systems.FRET based on orange (mOrange2 and red fluorescent protein (mKate2 partners allows the use of longer wavelength light, enabling deeper penetration of samples at lower energy and increased resolution with reduced back-ground auto-fluorescence. The FRET biosensors described in this paper for four new classes of compounds; (i cyclic polyols, (ii L-deoxy sugars, (iii β-linked disaccharides and (iv C4-dicarboxylates could be developed to study metabolism in vivo.

Fretting-corrosion behavior in hip implant modular junctions: The influence of friction energy and pH variation.

Science.gov (United States)

Royhman, Dmitry; Patel, Megha; Runa, Maria J; Wimmer, Markus A; Jacobs, Joshua J; Hallab, Nadim J; Mathew, Mathew T

2016-09-01

Recently, there has been increasing concern in the orthopedic community over the use of hip implant modular devices due to an increasing number of reports of early failure, failure that has been attributed to fretting-corrosion at modular interfaces. Much is still unknown about the electrochemical and mechanical degradation mechanisms associated with the use of such devices. Accordingly, the purpose of our study was to develop a methodology for testing the fretting-corrosion behavior of modular junctions. A fretting-corrosion apparatus was used to simulate the fretting-corrosion conditions of a CoCrMo hip implant head on a Ti6Al4V hip implant stem. The device features two perpendicularly-loaded CoCrMo pins that articulated against a Ti6Al4V rod. A sinusoidal fretting motion was applied to the rod at various displacement amplitudes (25, 50, 100, 150 and 200μm) at a constant load of 200N. Bovine calf serum at two different pH levels (3.0 and 7.6) was used to simulate the fluid environment around the joint. Experiments were conducted in two modes of electrochemical control - free-potential and potentiostatic. Electrochemical impedance spectroscopy tests were done before and after the fretting motion to assess changes in corrosion kinetics. In free potential mode, differences were seen in change in potential as a function of displacement amplitude. In general, VDrop (the drop in potential at the onset of fretting), VFretting, (the average potential during fretting), ΔVFretting (the change in potential from the onset of fretting to its termination) and VRecovery (the change in potential from the termination of fretting until stabilization) appeared linear at both pH levels, but showed drastic deviation from linearity at 100μm displacement amplitude. Subsequent EDS analysis revealed a large number of Ti deposits on the CoCrMo pin surfaces. Potentiostatic tests at both pH levels generally showed increasing current with increasing displacement amplitude. Electrochemical

Revisitation of FRET methods to measure intraprotein distances in Human Serum Albumin

Energy Technology Data Exchange (ETDEWEB)

Santini, S.; Bizzarri, A.R.; Cannistraro, S., E-mail: cannistr@unitus.it

2016-11-15

We revisited the FRET methods to measure the intraprotein distance between Trp-214 (used as donor) of Human Serum Albumin and its Cys-34, labelled with 1.5-Iaedans (used as acceptor). Variation of Trp fluorescence emission in terms of both intensity and lifetime, as well the enhancement of the acceptor fluorescence emission upon Trp excitation, have been monitored. A careful statistical analysis of the fluorescence results from ten independently prepared samples, combined with suitable spectral corrections, provided reproducible distances estimations by each one of the three methods. Even if monitoring of the donor lifetime variation in the presence of the acceptor reproduces at the best the crystallographic data, by allowing even sub-nanometre distance variations to be appreciated, we suggest that a comparative analysis of all the three methods, applied with statistical significance, should be preferred to achieve a better reliability of the FRET technique.

48-spot single-molecule FRET setup with periodic acceptor excitation

Science.gov (United States)

Ingargiola, Antonino; Segal, Maya; Gulinatti, Angelo; Rech, Ivan; Labanca, Ivan; Maccagnani, Piera; Ghioni, Massimo; Weiss, Shimon; Michalet, Xavier

2018-03-01

Single-molecule Förster resonance energy transfer (smFRET) allows measuring distances between donor and acceptor fluorophores on the 3-10 nm range. Solution-based smFRET allows measurement of binding-unbinding events or conformational changes of dye-labeled biomolecules without ensemble averaging and free from surface perturbations. When employing dual (or multi) laser excitation, smFRET allows resolving the number of fluorescent labels on each molecule, greatly enhancing the ability to study heterogeneous samples. A major drawback to solution-based smFRET is the low throughput, which renders repetitive measurements expensive and hinders the ability to study kinetic phenomena in real-time. Here we demonstrate a high-throughput smFRET system that multiplexes acquisition by using 48 excitation spots and two 48-pixel single-photon avalanche diode array detectors. The system employs two excitation lasers allowing separation of species with one or two active fluorophores. The performance of the system is demonstrated on a set of doubly labeled double-stranded DNA oligonucleotides with different distances between donor and acceptor dyes along the DNA duplex. We show that the acquisition time for accurate subpopulation identification is reduced from several minutes to seconds, opening the way to high-throughput screening applications and real-time kinetics studies of enzymatic reactions such as DNA transcription by bacterial RNA polymerase.

Förster Resonance Energy Transfer (FRET) from Triton X-100 to 4-benzothiazol-2-yl-phenol: Varying FRET efficiency with CMC of the donor (Triton X-100)

International Nuclear Information System (INIS)

Paul, Bijan Kumar; Ganguly, Aniruddha; Karmakar, Saswati; Guchhait, Nikhil

2013-01-01

A heterocyclic compound viz., 4-benzothiazol-2-yl-phenol (4B2YP) has been synthesized and its photophysics have been examined through steady-state absorption, emission and time resolved emission spectroscopic techniques, in brief. Then 4B2YP has been exploited as an acceptor in the Förster Resonance Energy Transfer (FRET) process from photoexcited benzene aromatic nucleus of Triton X-100 (TX-100) surfactant. Dependence of the energy transfer efficiency on the donor concentration with respect to its critical micelle concentration (CMC) is clearly reflected in the study. High values of Stern–Volmer constant (K SV ) for quenching of the donor fluorescence in the presence of the acceptor suggest the operation of long-range dipole–dipole interaction in the course of energy transfer process, while the inference is aptly supported from time resolved fluorescence decay results. Experimental results show maximum FRET efficiency at the CMC of the donor (TX-100). -- Highlights: • FRET from neutral surfactant Triton X-100 to chromophore 4-benzothiazol-2-yl-phenol. • Steady state and time resolved spectroscopy. • Long-range dipole–dipole interaction responsible for FRET. • FRET efficiency as a measure of CMC of surfactant

Förster Resonance Energy Transfer (FRET) from Triton X-100 to 4-benzothiazol-2-yl-phenol: Varying FRET efficiency with CMC of the donor (Triton X-100)

Energy Technology Data Exchange (ETDEWEB)

Paul, Bijan Kumar, E-mail: bijan.paul.chem.cu@gmail.com [Department of Chemistry, University of Calcutta, 92 A.P.C. Road, Calcutta 700009 (India); Ganguly, Aniruddha [Department of Chemistry, University of Calcutta, 92 A.P.C. Road, Calcutta 700009 (India); Karmakar, Saswati [Department of Chemistry, Sree Chaitanya College, Habra, North 24 Parganas (India); Guchhait, Nikhil, E-mail: nguchhait@yahoo.com [Department of Chemistry, University of Calcutta, 92 A.P.C. Road, Calcutta 700009 (India)

2013-11-15

A heterocyclic compound viz., 4-benzothiazol-2-yl-phenol (4B2YP) has been synthesized and its photophysics have been examined through steady-state absorption, emission and time resolved emission spectroscopic techniques, in brief. Then 4B2YP has been exploited as an acceptor in the Förster Resonance Energy Transfer (FRET) process from photoexcited benzene aromatic nucleus of Triton X-100 (TX-100) surfactant. Dependence of the energy transfer efficiency on the donor concentration with respect to its critical micelle concentration (CMC) is clearly reflected in the study. High values of Stern–Volmer constant (K{sub SV}) for quenching of the donor fluorescence in the presence of the acceptor suggest the operation of long-range dipole–dipole interaction in the course of energy transfer process, while the inference is aptly supported from time resolved fluorescence decay results. Experimental results show maximum FRET efficiency at the CMC of the donor (TX-100). -- Highlights: • FRET from neutral surfactant Triton X-100 to chromophore 4-benzothiazol-2-yl-phenol. • Steady state and time resolved spectroscopy. • Long-range dipole–dipole interaction responsible for FRET. • FRET efficiency as a measure of CMC of surfactant.

Effects of fretting fatigue on the residual stress of shot peened Ti-6Al-4V samples

International Nuclear Information System (INIS)

Martinez, S.A.; Sathish, S.; Blodgett, M.P.; Mall, S.; Namjoshi, S.

2005-01-01

X-ray diffraction residual stress measurement has been utilized as nondestructive tool for the characterization of fretting fatigue damage in shot peened samples of Ti-6Al-4V. Prior to fretting fatigue damage, compressive residual stresses were found to be uniform over the entire face of the sample and independent of the measurement direction. After fretting fatigue, inside and in the vicinity of the fretting damage zone large relaxation of compressive residual stress was observed. An anisotropic residual stress distribution has been observed in the fretting fatigue damaged region. Residual stress measurements in interrupted fretting fatigue experiments showed that the relaxation of residual stress increases as the number of fretting fatigue cycles increase. The results are discussed in the light of their importance in establishing X-ray diffraction residual stress measurement technique as a nondestructive tool to characterize fretting fatigue damage

Fuel-element vibration and bearing pad to pressure tube fretting

International Nuclear Information System (INIS)

Fisher, N.J.; Taylor, C.E.; Pettigrew, M.J.

1990-08-01

Fuel channel operation under boiling condition results in increased flow velocities, which may lead to unacceptable fuel-element vibration and bearing pad to pressure tube fretting. The existing endurance test database does not fully cover the range of future channel operating conditions. In particular, after refuelling, some channels for future designs may operate with two-phase flow conditions outside the range of endurance test conditions. Full-scale endurance testing at realistic steam-water conditions involves substantial energy costs. Therefore, fundamental laboratory investigations were conducted to define and endurance test matrix which adequately envelops the future range of operating conditions while minimizing both the number of tests and the energy requirement of individual tests. The main focus of the laboratory investigations was to establish the relationships between: fuel channel flow conditions and fuel-element vibration; and fuel-element vibration and bearing pad to pressure tube fretting. The vibration response of a single fuel element was measured over a wide range of operating conditions covering realistic fuel channel conditions and simulated endurance testing conditions. For higher void fractions, the vibration amplitudes measured in air/water were much higher than in steam/water, while for low void fractions, the amplitudes were similar. The measured amplitudes in steam/water varied very little over the range of temperature and pressure investigated. The effects of temperature, pressure tube oxide thickness, vibration amplitude and bearing pad manufacturer on pressure tube fretting were investigated. The fretting rate is extremely temperature dependent. For vibration amplitudes about three or four times greater than expected in-reactor conditions, peak fretting rates were observed in the 225 to 286 degrees C temperature range. Fretting rates were seven times less at the higher temperatures of 300 and 315 degrees C, and the lower temperatures

Influence of plasma molybdenizing and shot-peening on fretting damage behavior of titanium alloy

Energy Technology Data Exchange (ETDEWEB)

Tang, Chang-bin, E-mail: tcbtop@126.com [School of Metallurgy and Engineering, Xi’an University of Architecture and Technology, Xi’an, Shaanxi 710055 (China); Institute of Corrosion and Protection, Northwestern Polytechnical University, Xi’an, Shaanxi 710072 (China); Liu, Dao-xin, E-mail: liudaox@nwpu.edu.cn [Institute of Corrosion and Protection, Northwestern Polytechnical University, Xi’an, Shaanxi 710072 (China); Tang, Bin [Institute of Surface Engineering, Taiyuan University of Technology, Taiyuan Shanxi 030024 (China); Zhang, Xiao-hua [Institute of Corrosion and Protection, Northwestern Polytechnical University, Xi’an, Shaanxi 710072 (China); Qin, Lin [Institute of Surface Engineering, Taiyuan University of Technology, Taiyuan Shanxi 030024 (China); Liu, Cheng-song [Institute of Corrosion and Protection, Northwestern Polytechnical University, Xi’an, Shaanxi 710072 (China)

2016-12-30

Highlights: • Plasma molybdenizing increases FW resistance of Ti6Al4V, but reduces its FF life. • Shot-peened plasmamolybdenizing surface enhances FW and FF resistance of Ti6Al4V. • Combined treatment yields low surface-roughness & high hardness gradient distribution. • Combined treatment yields beneficial residual compressive stress & good toughness. • Anti-wear & -fatigue performance improvements for surface engineering applications. - Abstract: Effect of plasma molybdenizing and shot-peening on fretting wear and fretting fatigue behaviors of Ti6Al4V alloy was investigated. The plasma molybdenized layer composed of a dense molybdenum deposition layer and a Mo–Ti solid–solution layer can increase surface hardness by 2.8 times and cause its volume loss by fretting wear to decrease to 1/14 compared with that of the substrate. Plasma molybdenized treatment results in a significant decrease in resistance of the substrate to fretting fatigue. It is ascribed that the molybdenized layer with high hardness yields a low toughness, and its high surface roughness leads to a micro-notched effect. However, proper combination plasma molybdenizing and subsequent shot-peening may enhance the simultaneous fretting fatigue and fretting wear resistance of Ti6Al4V significantly, which can decrease the fretting wear volume loss to 1/27, and may increase the fretting fatigue life by more than 69 times. A synergistic improvement in fretting fatigue of the titanium alloy by combining surface alloying with shot-peening can be achieved. The results indicate that a beneficial residual compressive stress distribution, high surface hardness with suitable hardness gradient distribution, good apparent toughness, relatively low surface roughness, and excellent surface integrity are achieved.

Fretting and wear of stainless and ferritic steels in LMFBR steam generators

International Nuclear Information System (INIS)

Lewis, M.W.J.; Campbell, C.S.

1981-01-01

Steam generators for LMFBR's may be subject to both fretting wear as a result of flow-induced vibrations and to wear from larger amplitude sliding movements from thermal changes. Results of tests simulating the latter are given for stainless and ferritic steels. For the assessment of fretting wear damage, vibration assessments must be combined with data on specific wear rates. Test mechanisms used to study fretting in sodium covering impact, impact-slide and pure rubbing are described and results presented. (author)

MIL-L-87177 and CLT:X-10 Lubricants Improve Electrical Connector Fretting Corrosion Behavior

International Nuclear Information System (INIS)

AUKLAND, NEIL R.; HANLON, JAMES T.

1999-01-01

We have conducted a fretting research project using MIL-L-87177 and CLT: X-10 lubricants on Nano-miniature connectors. When they were fretted without lubricant, individual connectors first exceeded our 0.5 ohm failure criteria from 2,341 to 45,238 fretting cycles. With additional fretting, their contact resistance increased to more than 100,000 ohms. Unmodified MIL-L-87177 lubricant delayed the onset of first failure to between 430,000 and over 20,000,000 fretting cycles. MIL-L-87177 modified by addition of Teflon powder delayed first failure to beyond 5 million fretting cycles. Best results were obtained when Teflon was used and also when both the straight and modified lubricants were poured into and then out of the connector. CLT: X-10 lubricant delayed the onset of first failure to beyond 55 million cycles in one test where a failure was actually observed and to beyond 20 million cycles in another that was terminated without failure. CLT: X-10 recovered an unlubricated connector driven deeply into failure, with six failed pins recovering immediately and four more recovering during an additional 420 thousand fretting cycles. MIL-L-87177 was not able to recover a connector under similar conditions

Inferring properties of disordered chains from FRET transfer efficiencies

Science.gov (United States)

Zheng, Wenwei; Zerze, Gül H.; Borgia, Alessandro; Mittal, Jeetain; Schuler, Benjamin; Best, Robert B.

2018-03-01

Förster resonance energy transfer (FRET) is a powerful tool for elucidating both structural and dynamic properties of unfolded or disordered biomolecules, especially in single-molecule experiments. However, the key observables, namely, the mean transfer efficiency and fluorescence lifetimes of the donor and acceptor chromophores, are averaged over a broad distribution of donor-acceptor distances. The inferred average properties of the ensemble therefore depend on the form of the model distribution chosen to describe the distance, as has been widely recognized. In addition, while the distribution for one type of polymer model may be appropriate for a chain under a given set of physico-chemical conditions, it may not be suitable for the same chain in a different environment so that even an apparently consistent application of the same model over all conditions may distort the apparent changes in chain dimensions with variation of temperature or solution composition. Here, we present an alternative and straightforward approach to determining ensemble properties from FRET data, in which the polymer scaling exponent is allowed to vary with solution conditions. In its simplest form, it requires either the mean FRET efficiency or fluorescence lifetime information. In order to test the accuracy of the method, we have utilized both synthetic FRET data from implicit and explicit solvent simulations for 30 different protein sequences, and experimental single-molecule FRET data for an intrinsically disordered and a denatured protein. In all cases, we find that the inferred radii of gyration are within 10% of the true values, thus providing higher accuracy than simpler polymer models. In addition, the scaling exponents obtained by our procedure are in good agreement with those determined directly from the molecular ensemble. Our approach can in principle be generalized to treating other ensemble-averaged functions of intramolecular distances from experimental data.

FRET-Aptamer Assays for Bone Marker Assessment, C-Telopeptide, Creatinine, and Vitamin D

Science.gov (United States)

Bruno, John G.

2013-01-01

Astronauts lose 1.0 to 1.5% of their bone mass per month on long-duration spaceflights. NASA wishes to monitor the bone loss onboard spacecraft to develop nutritional and exercise countermeasures, and make adjustments during long space missions. On Earth, the same technology could be used to monitor osteoporosis and its therapy. Aptamers bind to targets against which they are developed, much like antibodies. However, aptamers do not require animal hosts or cell culture and are therefore easier, faster, and less expensive to produce. In addition, aptamers sometimes exhibit greater affinity and specificity vs. comparable antibodies. In this work, fluorescent dyes and quenchers were added to the aptamers to enable pushbutton, one-step, bind-and-detect fluorescence resonance energy transfer (FRET) assays or tests that can be freeze-dried, rehydrated with body fluids, and used to quantitate bone loss of vitamin D levels with a handheld fluorometer in the spacecraft environment. This work generated specific, rapid, one-step FRET assays for the bone loss marker C-telopeptide (CTx) when extracted from urine, creatinine from urine, and vitamin D congeners in diluted serum. The assays were quantified in nanograms/mL using a handheld fluorometer connected to a laptop computer to convert the raw fluorescence values into concentrations of each analyte according to linear standard curves. DNA aptamers were selected and amplified for several rounds against a 26- amino acid form of CTx, creatinine, and vitamin D. The commonalities between loop structures were studied, and several common loop structures were converted into aptamer beacons with a fluorophore and quencher on each end. In theory, when the aptamer beacon binds its cognate target (CTx bone peptide, creatinine, or vitamin D), it is forced open and no longer quenched, so it gives off fluorescent light (when excited) in proportion to the amount of target present in a sample. This proportional increase in fluorescence is

Influence of Fretting on Flexural Fatigue of 304 Stainless Steel and Mild Steel

National Research Council Canada - National Science Library

Bill, Robert

1978-01-01

Fretting fatigue experiments conducted on 304 stainless steel using a flexural-fatigue test arrangement with bolted-on fretting pads have demonstrated that fatigue life is reduced by at least a factor...

Characteristics of CANDU fuel bundles that caused pressure tube fretting at the bundle midplane

Energy Technology Data Exchange (ETDEWEB)

Dennier, D; Manzer, A M [Atomic Energy of Canada Ltd., Mississauga, ON (Canada); Koehn, E [Ontario Hydro, Toronto, ON (Canada)

1996-12-31

Detailed measurements on new bundles, and those that caused fretting during in- and out-reactor tests, have given insight into the factors responsible for fretting at the midplane of the inlet bundle. Bottom fuel elements that were attached near radial endplate spokes and had inboard bearing pads in the rolled joint cavity produced a significant portion of the observed fret marks. These elements are influenced by several driving forces that deflect the centre bearing pads towards the pressure tube surface. The evidence suggests that slight changes in bundle design may be possible to reduce pressure tube fretting. (author). 4 refs., 3 tabs., 8 figs.

Quantitative analysis chemistry

International Nuclear Information System (INIS)

Ko, Wansuk; Lee, Choongyoung; Jun, Kwangsik; Hwang, Taeksung

1995-02-01

This book is about quantitative analysis chemistry. It is divided into ten chapters, which deal with the basic conception of material with the meaning of analysis chemistry and SI units, chemical equilibrium, basic preparation for quantitative analysis, introduction of volumetric analysis, acid-base titration of outline and experiment examples, chelate titration, oxidation-reduction titration with introduction, titration curve, and diazotization titration, precipitation titration, electrometric titration and quantitative analysis.

A QM-MD simulation approach to the analysis of FRET processes in (bio)molecular systems. A case study: complexes of E. coli purine nucleoside phosphorylase and its mutants with formycin A.

Science.gov (United States)

Sobieraj, M; Krzyśko, K A; Jarmuła, A; Kalinowski, M W; Lesyng, B; Prokopowicz, M; Cieśla, J; Gojdź, A; Kierdaszuk, B

2015-04-01

Predicting FRET pathways in proteins using computer simulation techniques is very important for reliable interpretation of experimental data. A novel and relatively simple methodology has been developed and applied to purine nucleoside phosphorylase (PNP) complexed with a fluorescent ligand - formycin A (FA). FRET occurs between an excited Tyr residue (D*) and FA (A). This study aims to interpret experimental data that, among others, suggests the absence of FRET for the PNPF159A mutant in complex with FA, based on novel theoretical methodology. MD simulations for the protein molecule containing D*, and complexed with A, are carried out. Interactions of D* with its molecular environment are accounted by including changes of the ESP charges in S1, compared to S0, and computed at the SCF-CI level. FRET probability W F depends on the inverse six-power of the D*-A distance, R da . The orientational factor 0 < k(2) < 4 between D* and A is computed and included in the analysis. Finally W F is time-averaged over the MD trajectories resulting in its mean value. The red-shift of the tyrosinate anion emission and thus lack of spectral overlap integral and thermal energy dissipation are the reasons for the FRET absence in the studied mutants at pH 7 and above. The presence of the tyrosinate anion results in a competitive energy dissipation channel and red-shifted emission, thus in consequence in the absence of FRET. These studies also indicate an important role of the phenyl ring of Phe159 for FRET in the wild-type PNP, which does not exist in the Ala159 mutant, and for the effective association of PNP with FA. In a more general context, our observations point out very interesting and biologically important properties of the tyrosine residue in its excited state, which may undergo spontaneous deprotonation in the biomolecular systems, resulting further in unexpected physical and/or biological phenomena. Until now, this observation has not been widely discussed in the

Flow Cytometry Enables Multiplexed Measurements of Genetically Encoded Intramolecular FRET Sensors Suitable for Screening.

Science.gov (United States)

Doucette, Jaimee; Zhao, Ziyan; Geyer, Rory J; Barra, Melanie M; Balunas, Marcy J; Zweifach, Adam

2016-07-01

Genetically encoded sensors based on intramolecular FRET between CFP and YFP are used extensively in cell biology research. Flow cytometry has been shown to offer a means to measure CFP-YFP FRET; we suspected it would provide a unique way to conduct multiplexed measurements from cells expressing different FRET sensors, which is difficult to do with microscopy, and that this could be used for screening. We confirmed that flow cytometry accurately measures FRET signals using cells transiently transfected with an ERK activity reporter, comparing responses measured with imaging and cytometry. We created polyclonal long-term transfectant lines, each expressing a different intramolecular FRET sensor, and devised a way to bar-code four distinct populations of cells. We demonstrated the feasibility of multiplexed measurements and determined that robust multiplexed measurements can be conducted in plate format. To validate the suitability of the method for screening, we measured responses from a plate of bacterial extracts that in unrelated experiments we had determined contained the protein kinase C (PKC)-activating compound teleocidin A-1. The multiplexed assay correctly identifying the teleocidin A-1-containing well. We propose that multiplexed cytometric FRET measurements will be useful for analyzing cellular function and for screening compound collections. © 2016 Society for Laboratory Automation and Screening.

Directed Evolution to Engineer Monobody for FRET Biosensor Assembly and Imaging at Live-Cell Surface.

Science.gov (United States)

Limsakul, Praopim; Peng, Qin; Wu, Yiqian; Allen, Molly E; Liang, Jing; Remacle, Albert G; Lopez, Tyler; Ge, Xin; Kay, Brian K; Zhao, Huimin; Strongin, Alex Y; Yang, Xiang-Lei; Lu, Shaoying; Wang, Yingxiao

2018-04-19

Monitoring enzymatic activities at the cell surface is challenging due to the poor efficiency of transport and membrane integration of fluorescence resonance energy transfer (FRET)-based biosensors. Therefore, we developed a hybrid biosensor with separate donor and acceptor that assemble in situ. The directed evolution and sequence-function analysis technologies were integrated to engineer a monobody variant (PEbody) that binds to R-phycoerythrin (R-PE) dye. PEbody was used for visualizing the dynamic formation/separation of intercellular junctions. We further fused PEbody with the enhanced CFP and an enzyme-specific peptide at the extracellular surface to create a hybrid FRET biosensor upon R-PE capture for monitoring membrane-type-1 matrix metalloproteinase (MT1-MMP) activities. This biosensor revealed asymmetric distribution of MT1-MMP activities, which were high and low at loose and stable cell-cell contacts, respectively. Therefore, directed evolution and rational design are promising tools to engineer molecular binders and hybrid FRET biosensors for monitoring molecular regulations at the surface of living cells. Copyright © 2018 Elsevier Ltd. All rights reserved.

Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting

OpenAIRE

Bajar, Bryce T.; Wang, Emily S.; Lam, Amy J.; Kim, Bongjae B.; Jacobs, Conor L.; Howe, Elizabeth S.; Davidson, Michael W.; Lin, Michael Z.; Chu, Jun

2016-01-01

Many genetically encoded biosensors use F?rster resonance energy transfer (FRET) to dynamically report biomolecular activities. While pairs of cyan and yellow fluorescent proteins (FPs) are most commonly used as FRET partner fluorophores, respectively, green and red FPs offer distinct advantages for FRET, such as greater spectral separation, less phototoxicity, and lower autofluorescence. We previously developed the green-red FRET pair Clover and mRuby2, which improves responsiveness in intra...

MnO2 nanosheet mediated "DD-A" FRET binary probes for sensitive detection of intracellular mRNA.

Science.gov (United States)

Ou, Min; Huang, Jin; Yang, Xiaohai; Quan, Ke; Yang, Yanjing; Xie, Nuli; Wang, Kemin

2017-01-01

The donor donor-acceptor (DD-A) FRET model has proven to have a higher FRET efficiency than donor-acceptor acceptor (D-AA), donor-acceptor (D-A), and donor donor-acceptor acceptor (DD-AA) FRET models. The in-tube and in-cell experiments clearly demonstrate that the "DD-A" FRET binary probes can indeed increase the FRET efficiency and provide higher imaging contrast, which is about one order of magnitude higher than the ordinary "D-A" model. Furthermore, MnO 2 nanosheets were employed to deliver these probes into living cells for intracellular TK1 mRNA detection because they can adsorb ssDNA probes, penetrate across the cell membrane and be reduced to Mn 2+ ions by intracellular GSH. The results indicated that the MnO 2 nanosheet mediated "DD-A" FRET binary probes are capable of sensitive and selective sensing gene expression and chemical-stimuli changes in gene expression levels in cancer cells. We believe that the MnO 2 nanosheet mediated "DD-A" FRET binary probes have the potential as a simple but powerful tool for basic research and clinical diagnosis.

An ad-hoc fretting wear tribotester design for thin steel wires

Directory of Open Access Journals (Sweden)

Llavori Iñigo

2018-01-01

Full Text Available Steel wire ropes experience fretting wear damage when the rope runs over a sheave promoting an oscillatory motion between the wires. Consequently, wear scars appear between the contacting wires leading to an increase of the stress field and the following rupture of the wires due to fatigue. That is why the understanding and prediction of the fretting wear phenomena of thin wires is fundamental in order to improve the performance of steel wire ropes. The present research deals with the design of an ad-hoc fretting wear test machine for thin wires. The test apparatus is designed for testing thin wires with a maximum diameter of 1.0 mm, at slip amplitudes ranging from 5 to 300 μm, crossing angle between 0-90°, and contacting force ranging from 0,5 to 5 N. The working principle of displacement amplitude and contacting force as well as the crossing angle between the wires are described. Preliminary studies for understanding the fretting wear characteristics are presented, analysing 0.45 mm diameter cold-drawn eutectoid carbon steel (0.8% C wires (tensile strength higher than 3000 MPa.

Single-molecule three-color FRET with both negligible spectral overlap and long observation time.

Directory of Open Access Journals (Sweden)

Sanghwa Lee

Full Text Available Full understanding of complex biological interactions frequently requires multi-color detection capability in doing single-molecule fluorescence resonance energy transfer (FRET experiments. Existing single-molecule three-color FRET techniques, however, suffer from severe photobleaching of Alexa 488, or its alternative dyes, and have been limitedly used for kinetics studies. In this work, we developed a single-molecule three-color FRET technique based on the Cy3-Cy5-Cy7 dye trio, thus providing enhanced observation time and improved data quality. Because the absorption spectra of three fluorophores are well separated, real-time monitoring of three FRET efficiencies was possible by incorporating the alternating laser excitation (ALEX technique both in confocal microscopy and in total-internal-reflection fluorescence (TIRF microscopy.

EXPERIMENTAL INVESTIGTION OF THE FRETTING PHENOMENON-DEPENDENCE OF NUMBERS CYCLES

Directory of Open Access Journals (Sweden)

Ştefan GHIMISI

2014-12-01

Full Text Available The present paper argues that adhesion forces and elastic deformation in the contact zone may contribute significantly to the relative displacement during fretting of metals. A simultaneously applied tangential force and normal into contact appears a adhesion force. A tangential force whose magnitude is less equal on greater than the force of limiting friction will not give rise on give rise to a sliding motion.It is determined the energy loss dissipated per fretting cycle.

Fretting friction and wear characteristics of magnetorheological fluid under different magnetic field strengths

International Nuclear Information System (INIS)

Zhang, P.; Lee, K.H.; Lee, C.H.

2017-01-01

A magnetorheological fluid (MRF) performs differently under different magnetic field strength. This study examined the fretting friction and wear characteristics of MRFs under a range of magnetic field strengths and oscillation frequencies. The fretting friction and wear behaviors of MRF are investigated using a fretting friction and wear tester. The surfaces of specimen are examined by optical microscopy and 3D surface profilometer before and after the tests and wear surface profiles, the wear volume loss and wear coefficient for each magnetic field strength are evaluated. The results show that the friction and wear properties of MRF change according to the magnetic field strength and oscillation frequency. - Highlights: • Fretting friction and wear characteristics of MRF is examined. • The friction coefficients increased with increasing magnetic field strength. • The coefficient of friction decreased with increasing oscillation frequency. • Wear volume and coefficient become worse with increasing magnetic field strength.

Fretting and Corrosion Damage in Taper Adapter Sleeves for Ceramic Heads: A Retrieval Study.

Science.gov (United States)

MacDonald, Daniel W; Chen, Antonia F; Lee, Gwo-Chin; Klein, Gregg R; Mont, Michael A; Kurtz, Steven M; Cates, Harold E; Kraay, Matthew J; Rimnac, Clare M

2017-09-01

During revision surgery with a well-fixed stem, a titanium sleeve can be used in conjunction with a ceramic head to achieve better stress distribution across the taper surface. In vitro testing suggests that corrosion is not a concern in sleeved ceramic heads; however, little is known about the in vivo fretting corrosion of the sleeves. The purpose of this study was to investigate fretting corrosion in sleeved ceramic heads in retrieved total hip arthroplasties. Thirty-seven sleeved ceramic heads were collected during revision. The femoral heads and sleeves were implanted 0.0-3.3 years. The implants were revised predominantly for instability, infection, and loosening. Fifty percent of the retrievals were implanted during a primary surgery. Fretting corrosion was assessed using the Goldberg-Higgs semiquantitative scoring system. Mild-to-moderate fretting corrosion scores (score = 2-3) were observed in 92% of internal tapers, 19% of external tapers, and 78% of the stems. Severe fretting corrosion was observed in 1 stem trunnion that was previously retained during revision surgery and none of the retrieved sleeves. There was no difference in corrosion damage of sleeves used in primary or revision surgery. The fretting corrosion scores in this study were predominantly mild and lower than reported fretting scores of cobalt-chrome heads in metal-on-polyethylene bearings. Although intended for use in revisions, we found that the short-term in vivo corrosion behavior of the sleeves was similar in both primary and revision surgery applications. From an in vivo corrosion perspective, sleeves are a reasonable solution for restoring the stem taper during revision surgery. Copyright © 2017. Published by Elsevier Inc.

Experimental and Numerical Investigations of Fretting Fatigue Behavior for Steel Q235 Single-Lap Bolted Joints

Directory of Open Access Journals (Sweden)

Yazhou Xu

2016-01-01

Full Text Available This work aims to investigate the fretting fatigue life and failure mode of steel Q235B plates in single-lap bolted joints. Ten specimens were prepared and tested to fit the S-N curve. SEM (scanning electron microscope was then employed to observe fatigue crack surfaces and identify crack initiation, crack propagation, and transient fracture zones. Moreover, a FEM model was established to simulate the stress and displacement fields. The normal contact stress, tangential contact stress, and relative slipping displacement at the critical fretting zone were used to calculate FFD values and assess fretting fatigue crack initiation sites, which were in good agreement with SEM observations. Experimental results confirmed the fretting fatigue failure mode for these specimens. It was found that the crack initiation resulted from wear regions at the contact surfaces between plates, and fretting fatigue cracks occurred at a certain distance away from hole edges. The proposed FFD-N relationship is an alternative approach to evaluate fretting fatigue life of steel plates in bolted joints.

Fretting wear characteristic tests of X2-GEN midgrid for SMART under a FIV rod trace

Energy Technology Data Exchange (ETDEWEB)

Lee, Young Ho; Lee, Kang Hee; Kim, Jae Yong; Kim, Hyung Kyu [KAERI, Daejeon (Korea, Republic of)

2011-12-15

The KEPCO Nuclear Fuel Co. requested the fretting wear characteristic tests of a X2-GEN midgrid under a FIV rod trace at room temperature air. The following results were obtained for the fretting wear test. {center_dot} Fretting wear tests under a FIV rod trace Based on the result of the fretting wear tests of the X2-GEN and 17ACE7 1x1 mid-grid under a FIV rod trace, X2-GEN mid-grid showed a slightly severe wear volume rather than 17ACE7 spring. But, maximum wear depth shows an opposite behavior. This is due to spring shape effect. The fretting wear mechanisms at each mid-grid were influenced by each spring shape, that are depended on the different impacting behavior under a FIV rod motion. Up to 5x105 cycles, wear characteristics of each mid-grid shows a relatively similar wear rate. Consequently, it is necessary to further study for examining exact fretting wear behavior under a FIV rod tra

Homo-FRET imaging as a tool to quantify protein and lipid clustering.

Science.gov (United States)

Bader, Arjen N; Hoetzl, Sandra; Hofman, Erik G; Voortman, Jarno; van Bergen en Henegouwen, Paul M P; van Meer, Gerrit; Gerritsen, Hans C

2011-02-25

Homo-FRET, Förster resonance energy transfer between identical fluorophores, can be conveniently measured by observing its effect on the fluorescence anisotropy. This review aims to summarize the possibilities of fluorescence anisotropy imaging techniques to investigate clustering of identical proteins and lipids. Homo-FRET imaging has the ability to determine distances between fluorophores. In addition it can be employed to quantify cluster sizes as well as cluster size distributions. The interpretation of homo-FRET signals is complicated by the fact that both the mutual orientations of the fluorophores and the number of fluorophores per cluster affect the fluorescence anisotropy in a similar way. The properties of the fluorescence probes are very important. Taking these properties into account is critical for the correct interpretation of homo-FRET signals in protein- and lipid-clustering studies. This is be exemplified by studies on the clustering of the lipid raft markers GPI and K-ras, as well as for EGF receptor clustering in the plasma membrane. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Experimental fretting-wear studies of steam generator materials

International Nuclear Information System (INIS)

Fisher, N.J.; Chow, A.B.; Weckwerth, M.K.

1994-01-01

Flow-induced vibration of steam generator tubes results in fretting-wear damage due to impacting and rubbing of the tubes against their supports. This damage can be predicted by computing tube response to flow-induced excitation forces using analytical techniques, and then relating this response to resultant wear damage using experimentally-derived wear coefficients. Fretting-wear of steam generator materials has been studied experimentally at Chalk River Laboratories for two decades. Tests are conducted in machines that simulate steam generator environmental conditions and tube-to-support dynamic interactions. Different tube and support materials, tube-to-support clearances and tube support geometries have been studied. As well, the effect of environmental conditions, such as temperature, oxygen content, pH and chemistry control additive, have been investigated. Early studies showed that damage was related to contact force as long as other parameters, such as geometry and motion were held constant. Later studies have shown that damage is related to a parameter called work-rate, which combines both contact force and sliding distance. Results of short- and long-term fretting-wear tests for CANDU steam generator materials at realistic environmental conditions are presented. These results demonstrate that work-rate is appropriate correlating parameter for impact-sliding interaction

Aspects of fretting wear of sprayed cermet coatings

International Nuclear Information System (INIS)

Chivers, T.C.

1985-01-01

Two experimental fretting programmes which investigated aspects of fretting wear of sprayed cermet coatings are reviewed. These programmes were conducted in support of components used in the advanced gas-cooled reactor. It is speculated that the results from these programmes are compatible with a simple two-stage wear model. This model assumes that an initial wear process occurs which is dominated by an interlocking and removal of asperities. Such a phase will be dependent on the superficial contact areas and possibly the interfacial load, but the latter aspect is not considered. This initial wear is of very short duration and is followed by a mild, oxidative, wear mode. Coatings data are also compared with those for structural steels. In short-term low temperature tests it appears that structural steels have comparable performance with the cermet coatings but it is argued that this is an artefact of the wear process. However, at high temperatures (600 0 C) wear of stainless steel could not be determined, the specimens showing a net weight gain. It is concluded that for in-reactor fretting applications cermet coatings will have advantages over structural steels at low temperatures. Even in high temperature regions some operation at low temperatures is experienced and consequently cermet coatings may be useful here also. (orig.)

Forster Resonance Energy Transfer (FRET) Analysis of Dual CFP/YFP Labeled AMPA Receptors Reveals Structural Rearrangement within the C-Terminal Domain during Receptor Activation

DEFF Research Database (Denmark)

Zachariassen, Linda Grønborg; Katchan, Mila; Plested, Andrew

2014-01-01

that retain function and display intrareceptor FRET. This includes a construct (GluA2-6Y-10C) containing YFP in the intracellular loop between the M1 and M2 membrane-embedded segments and CFP inserted in the C-ter- minal domain (CTD). GluA2-6Y-10C displays FRET with an efficiency of 0.11 while retaining wild......-type receptor expression and kinetic properties. We have used GluA2-6Y-10C to study conformational changes in homomeric GluA2 receptors during receptor activation. Our results show that the FRET efficiency is dependent on functional state of GluA2-6Y-10C and hereby indi- cates that the intracellular CTD...

Molecularly imprinted fluorescent probe based on FRET for selective and sensitive detection of doxorubicin

Energy Technology Data Exchange (ETDEWEB)

Xu, Zhifeng, E-mail: 897061147@qq.com [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Deng, Peihong; Li, Junhua [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China); Xu, Li [Department of Applied Chemistry, College of Materials and Energy, South China Agricultural University, Guangzhou 510642 (China); Tang, Siping [College of Chemistry and Materials Science, Hengyang Normal University, Key Laboratory of Functional Organometallic Materials of Hunan Province University, Hengyang 421008 (China)

2017-04-15

Highlights: • FRET-based molecularly imprinted probe for detection of doxorubicin was prepared. • The detection limit of the probe was 13.8 nM for doxorubicin. • The FRET-based probe had a higher selectivity for the template than ordinary MIMs. - Abstract: In this work, a new type of fluorescent probe for detection of doxorubicin has been constructed by the combined use of fluorescence resonance energy transfer (FRET) technology and molecular imprinting technique (MIT). Using doxorubicin as the template, the molecularly imprinted polymer thin layer was fabricated on the surfaces of carbon dot (CD) modified silica by sol-gel polymerization. The excitation energy of the fluorescent donor (CDs) could be transferred to the fluorescent acceptor (doxorubicin). The FRET based fluorescent probe demonstrated high sensitivity and selectivity for doxorubicin. The detection limit was 13.8 nM. The fluorescent probe was successfully applied for detecting doxorubicin in doxorubicin-spiked plasmas with a recovery of 96.8–103.8%, a relative standard deviation (RSD) of 1.3–2.8%. The strategy for construction of FRET-based molecularly imprinted materials developed in this work is very promising for analytical applications.

Rational design of FRET-based sensor proteins

NARCIS (Netherlands)

Merkx, M.

2008-01-01

Real-time imaging of molecular events inside living cells is important for understanding the basis of physiological processes and diseases. Genetically encoded sensors that use fluorescence resonance energy transfer (FRET) between two fluorescent proteins are attractive in this respect because they

A dansyl-rhodamine chemosensor for Fe(III) based on off-on FRET

Science.gov (United States)

Piao, Jingyu; Lv, Jia; Zhou, Xin; Zhao, Tong; Wu, Xue

2014-07-01

A novel fluorescent chemosensor bearing a rhodamine and a dansyl moiety was developed for highly selective detection of Fe3+ based on fluorescence resonance energy transfer (FRET) mechanism. Binding of Fe3+ to the chemosensor induced spirolactam ring opening in the rhodamine moiety and subsequent off-on FRET from the dansyl energy donor to the rhodamine energy acceptor due to the spectral overlap between the emission of the dansyl moiety and the absorption of the ring opened rhodamine moiety. Job's plot analysis indicated a 1:1 binding stoichiometry between the chemosensor and Fe3+. The association constant was estimated to be 2.72 × 103 M-1 according to the Benesi-Hildebrand method. With the feature of easy synthesis, simple structural skeleton and excellent sensing ability, the newly synthesized chemosensor provided the potential for applying as a highly selective fluorescent probe in complex samples containing various competitive metal ions and developing other metal ion chemosensors to fulfill various needs of biological and environmental field.

Residual stress relaxation due to fretting fatigue in shot peened surfaces of Ti-6Al-4V

International Nuclear Information System (INIS)

Martinez, S.A.; Blodgett, M.P.; Mall, S.; Sathish, S.; Namjoshi, S.

2003-01-01

Fretting fatigue occurs at locations where the materials are sliding against each other under load. In order to enhance the fatigue life under fretting conditions the surface of the component is shot peened. In general, the shot peening process produces a compressive stress on the surface of the material, thereby increasing the resistance of the material to crack initiation. This paper presents the relaxation of residual stress caused during fretting fatigue. X-ray diffraction has been utilized as the method to measure residual stress in fretting fatigued samples of Ti-6Al-4V

Approximate stresses in 2-D flat elastic contact fretting problems

Science.gov (United States)

Urban, Michael Rene

Fatigue results from the cyclic loading of a solid body. If the body subject to fatigue is in contact with another body and relative sliding motion occurs between these two bodies, then rubbing surface damage can accelerate fatigue failure. The acceleration of fatigue failure is especially important if the relative motion between the two bodies results in surface damage without excessive surface removal via wear. The situation just described is referred to as fretting fatigue. Understanding of fretting fatigue is greatly enhanced if the stress state associated with fretting can be characterized. For Hertzian contact, this can readily be done. Unfortunately, simple stress formulae are not available for flat body contact. The primary result of the present research is the development of a new, reasonably accurate, approximate closed form expression for 2-dimensional contact stresses which has been verified using finite element modeling. This expression is also combined with fracture mechanics to provide a simple method of determining when a crack is long enough to no longer be affected by the contact stress field. Lower bounds on fatigue life can then easily be calculated using fracture mechanics. This closed form expression can also be used to calculate crack propagation within the contact stress field. The problem of determining the cycles required to generate an initial crack and what to choose as an initial crack size is unresolved as it is in non-fretting fatigue.

A Study on Surface Modification of Al7075-T6 Alloy against Fretting Fatigue Phenomenon

Directory of Open Access Journals (Sweden)

E. Mohseni

2014-01-01

Full Text Available Aircraft engines, fuselage, automobile parts, and energy saving strategies in general have promoted the interest and research in the field of lightweight materials, typically on alloys based on aluminum. Aluminum alloy itself does not have suitable wear resistance; therefore, it is necessary to enhance surface properties for practical applications, particularly when aluminum is in contact with other parts. Fretting fatigue phenomenon occurs when two surfaces are in contact with each other and one or both parts are subjected to cyclic load. Fretting drastically decreases the fatigue life of materials. Therefore, investigating the fretting fatigue life of materials is an important subject. Applying surface modification methods is anticipated to be a supreme solution to gradually decreasing fretting damage. In this paper, the authors would like to review methods employed so far to diminish the effect of fretting on the fatigue life of Al7075-T6 alloy. The methods include deep rolling, shot peening, laser shock peening, and thin film hard coatings. The surface coatings techniques are comprising physical vapor deposition (PVD, hard anodizing, ion-beam-enhanced deposition (IBED, and nitriding.

FRET-based modified graphene quantum dots for direct trypsin quantification in urine

Energy Technology Data Exchange (ETDEWEB)

Poon, Chung-Yan; Li, Qinghua [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong); Zhang, Jiali; Li, Zhongping [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong); Research Center of Environmental Science and Engineering, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Dong, Chuan [Research Center of Environmental Science and Engineering, School of Chemistry and Chemical Engineering, Shanxi University, Taiyuan 030006 (China); Lee, Albert Wai-Ming; Chan, Wing-Hong [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong); Li, Hung-Wing, E-mail: hwli@hkbu.edu.hk [Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong Special Administrative Region (Hong Kong)

2016-04-21

A versatile nanoprobe was developed for trypsin quantification with fluorescence resonance energy transfer (FRET). Here, fluorescence graphene quantum dot is utilized as a donor while a well-designed coumarin derivative, CMR2, as an acceptor. Moreover, bovine serum albumin (BSA), as a protein model, is not only served as a linker for the FRET pair, but also a fluorescence enhancer of the quantum dots and CMR2. In the presence of trypsin, the FRET system would be destroyed when the BSA is digested by trypsin. Thus, the emission peak of the donor is regenerated and the ratio of emission peak of donor/emission peak of acceptor increased. By the ratiometric measurement of these two emission peaks, trypsin content could be determined. The detection limit of trypsin was found to be 0.7 μg/mL, which is 0.008-fold of the average trypsin level in acute pancreatitis patient's urine suggesting a high potential for fast and low cost clinical screening. - Highlights: • A FRET-based biosensor was developed for direct quantification of trypsin. • Fast and sensitive screening of pancreatic disease was facilitated. • The direct quantification of trypsin in urine samples was demonstrated.

Fretting-wear damage of heat exchanger tubes: a proposed damage criterion based on tube vibration response

International Nuclear Information System (INIS)

Yetisir, M.; McKerrow, E.; Pettigrew, M.J.

1997-01-01

A simple criterion is proposed to estimate fretting-wear damage in heat exchanger tubes with clearance supports. The criterion is based on parameters such as vibration frequency, mid-span vibration amplitude, span length, tube mass and an empirical wear coefficient. It is generally accepted that fretting-wear damage is proportional to a parameter called work-rate. Work-rate is a measure of the dynamic interaction between a vibrating tube and its supports. Due to the complexity of the impact-sliding behavior at the clearance-supports, work-rate calculations for heat exchanger tubes require specialized non-linear finite element codes. These codes include contact models for various clearance-support geometries. Such non-linear finite element analyses are complex, expensive and time consuming. The proposed criterion uses the results of linear vibration analysis (i.e., vibration frequency and mid-span vibration amplitude due to turbulence) and does not require a non-linear analysis. It can be used by non-specialists for a quick evaluation of the expected work-rate, and hence, the fretting-wear damage of heat exchanger tubes. The proposed criterion was obtained from an extensive parametric study that was conducted using a non-linear finite element program. It is shown that, by using the proposed work-rate criteria, work-rate can be estimated within a factor of two. This result, however, requires further testing with more complicated flow patterns. (author)

Surveillance of siRNA integrity by FRET imaging

Science.gov (United States)

Järve, Anne; Müller, Julius; Kim, Il-Han; Rohr, Karl; MacLean, Caroline; Fricker, Gert; Massing, Ulrich; Eberle, Florian; Dalpke, Alexander; Fischer, Roger; Trendelenburg, Michael F.; Helm, Mark

2007-01-01

Techniques for investigation of exogenous small interfering RNA (siRNA) after penetration of the cell are of substantial interest to the development of efficient transfection methods as well as to potential medical formulations of siRNA. A FRET-based visualization method including the commonplace dye labels fluorescein and tetramethylrhodamin (TMR) on opposing strands of siRNA was found compatible with RNA interference (RNAi). Investigation of spectral properties of three labelled siRNAs with differential FRET efficiencies in the cuvette, including pH dependence and FRET efficiency in lipophilic environments, identified the ratio of red and green fluorescence (R/G-ratio) as a sensitive parameter, which reliably identifies samples containing >90% un-degraded siRNA. Spectral imaging of siRNAs microinjected into cells showed emission spectra indistinguishable from those measured in the cuvette. These were used to establish a calibration curve for assessing the degradation state of siRNA in volume elements inside cells. An algorithm, applied to fluorescence images recorded in standard green and red fluorescence channels, produces R/G-ratio images of high spatial resolution, identifying volume elements in the cell with high populations of intact siRNA with high fidelity. To demonstrate the usefulness of this technique, the movement of intact siRNA molecules are observed after introduction into the cytosol by microinjection, standard transfection and lipofection with liposomes. PMID:17890733

In vitro simulation of fretting-corrosion in hip implant modular junctions: The influence of pH.

Science.gov (United States)

Royhman, Dmitry; Patel, Megha; Jacobs, Joshua J; Wimmer, Markus A; Hallab, Nadim J; Mathew, Mathew T

2018-02-01

The fretting-corrosion behavior of mixed metal contacts is affected by various mechanical and electrochemical parameters. Crevice conditions at the junction and patient-specific pathologies can affect the pH of the prosthetic environment. The main objective of this study is to understand the effect of pH variation at the stem/head junction of the hip implant under fretting corrosion exposure. We hypothesized that pH will have a significant influence on the fretting-corrosion behavior hip implant modular junctions. A custom-made setup was used to evaluate the fretting corrosion behavior of hip implant modular junctions. A Newborn calf serum solution (30 g/L protein content) was used to simulate the synovial fluid environment. A sinusoidal fretting motion, with a displacement amplitude of +50 µm, was applied to the Ti alloy rod. The effects of pathology driven, periprosthetic pH variation were simulated at four different pH levels (3.0, 4.5, 6.0 and 7.6). Electrochemical and mechanical properties were evaluated before, during, and after the applied fretting motion. The impedance of the system was increased in response to the fretting motion. The hysteresis tangential load/displacement behavior was not affected by pH level. The worn surfaces of CoCrMo pins exhibited the presence of tribolayer or organic deposits, in the pH 4.5 group, which may explain the lower drop in potential and mass loss observed in that group. Mechanically dominated wear mechanisms, namely, adhesive wear was shown in the pH 7.6 group, which may account for a higher potential drop and metal content loss. This study suggests that the fretting-corrosion mechanisms in hip implant are affected by the pH levels of the surrounding environment and patient-specific factors. Copyright © 2017. Published by Elsevier Ltd.

Hyperspectral imaging for simultaneous measurements of two FRET biosensors in pancreatic β-cells.

Science.gov (United States)

Elliott, Amicia D; Bedard, Noah; Ustione, Alessandro; Baird, Michelle A; Davidson, Michael W; Tkaczyk, Tomasz; Piston, David W

2017-01-01

Fluorescent protein (FP) biosensors based on Förster resonance energy transfer (FRET) are commonly used to study molecular processes in living cells. There are FP-FRET biosensors for many cellular molecules, but it remains difficult to perform simultaneous measurements of multiple biosensors. The overlapping emission spectra of the commonly used FPs, including CFP/YFP and GFP/RFP make dual FRET measurements challenging. In addition, a snapshot imaging modality is required for simultaneous imaging. The Image Mapping Spectrometer (IMS) is a snapshot hyperspectral imaging system that collects high resolution spectral data and can be used to overcome these challenges. We have previously demonstrated the IMS's capabilities for simultaneously imaging GFP and CFP/YFP-based biosensors in pancreatic β-cells. Here, we demonstrate a further capability of the IMS to image simultaneously two FRET biosensors with a single excitation band, one for cAMP and the other for Caspase-3. We use these measurements to measure simultaneously cAMP signaling and Caspase-3 activation in pancreatic β-cells during oxidative stress and hyperglycemia, which are essential components in the pathology of diabetes.

An experimental study on the key fretting variables for flexible marine risers

OpenAIRE

O’Halloran, S.M.; Harte, A.M.; Shipway, P.H.; Leen, S.B.

2018-01-01

This paper presents an experimental investigation into the effects of contact conformity, contact pressure and displacement amplitude on the gross-slip fretting behaviour grease-lubricated cylinder-on-flat contacts in the context of flexible marine riser pressure armour wire, and compares behaviour with that observed in unlubricated conditions. Characterisation of friction and wear is critical to fretting fatigue life prediction in flexible risers since friction directly controls trailing-edg...

Effect of mixed alloy combinations on fretting corrosion performance of spinal screw and rod implants.

Science.gov (United States)

Mali, Sachin A; Singh, Vaneet; Gilbert, Jeremy L

2017-07-01

Spinal implants are made from a variety of materials to meet the unique mechanical demands of each application. However, the medical device community has raised concern about mixing dissimilar metals in an implant because of fear of inducing corrosion. There is a lack of systematic studies on the effects of mixing metals on performance of spinal implants, especially in fretting corrosion conditions. Hence, the goal was to determine whether mixing stainless steel (SS316L), titanium alloy (Ti6Al4V) and cobalt chromium (CoCrMo) alloy components in a spinal implant leads to any increased risk of corrosion degradation. Spinal constructs consisting of single assembly screw-connector-rod components were tested using a novel short-term cyclic fretting corrosion test method. A total of 17 alloy component combinations (comprised of SS316L, Ti6Al4V-anodized and CoCrMo alloy for rod, screws and connectors) were tested under three anatomic orientations. Spinal constructs having all SS316L were most susceptible to fretting-initiated crevice corrosion attack and showed higher average fretting currents (∼25 - 30 µA), whereas constructs containing all Ti6Al4V components were less susceptible to fretting corrosion with average fretting currents in the range of 1 - 6 µA. Mixed groups showed evidence of fretting corrosion but they were not as severe as all SS316L group. SEM results showed evidence of severe corrosion attack in constructs having SS316L components. There also did not appear to be any galvanic effects of combining alloys together. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 1169-1177, 2017. © 2016 Wiley Periodicals, Inc.

Combined effects of fretting and pollutant particles on the contact resistance of the electrical connectors

Directory of Open Access Journals (Sweden)

Zhigang Kong

2017-06-01

Full Text Available Usually, when electrical connectors operate in vibration environments, fretting will be produced at the contact interfaces. In addition, serious environmental pollution particles will affect contact resistance of the connectors. The fretting will worsen the reliability of connectors with the pollutant particles. The combined effects of fretting and quartz particles on the contact resistance of the gold plating connectors are studied with a fretting test system. The results show that the frequencies have obvious effect on the contact resistance. The higher the frequency, the higher the contact resistance is. The quartz particles cause serious wear of gold plating, which make the nickel and copper layer exposed quickly to increase the contact resistance. Especially in high humidity environments, water supply certain adhesion function and make quartz particles easy to insert or cover the contact surfaces, and even cause opening resistance.

The Necessity of a New Type Test Rig for the Development of an Evaluation Method in Grid Fretting Problems

International Nuclear Information System (INIS)

Lee, Young-Ho; Kim, Hyung-Kyu

2007-01-01

A grid fretting problem is recognized as one of the most important degradation mechanisms even though the examination results of fretting experiments could be applied to the development and design of spacer grid structures. This is because it is difficult to develop a fretting wear model for a grid fretting problem due to the various wear mechanisms involved according to the mechanical and environmental variables, the contact condition with a spring/dimple and the material properties. A number of spring shapes has been developed in KAERI and their performance tests such as fretting wear, flow-induced vibration (FIV) tests, etc. have been carried out from a part unit to a full assembly scale. From the unit part fretting test results, one of the noticeable results is that the contacting force (normal load) was gradually decreased with increasing number of fretting cycles due to a depth increase and this behavior was closely related to the contacting spring shape. When considering the actual contact condition between a fuel rod and a spring/dimple, if a fretting wear progresses due to a FIV under a specific normal load exerted on the fuel rod by an elastic deformation of the spring, the contacting force between the fuel rod and dimple that are located in the opposite side should be decreased. Consequently, an evaluation of developed spacer grids against fretting wear damage should be performed with the results of 1x1 cell unit experiments because a contacting force is one of the most important variables that influences a fretting wear mechanism. The discussion was focused on the development procedure of a new test rig and its performance by using a 1x1 cell unit test rig. (authors)

An overview of the Canadian program to investigate vibration and fretting in nuclear fuel assemblies

International Nuclear Information System (INIS)

Oldaker, I.E.; Lane, A.D.; Forrest, C.F.

The development of a model that would allow the fuel designer to predict the occurrence of fretting could materially reduce the amount of development testing of a new fuel design. To achieve this, we are working in several areas: to identify and measure the phenomena that excite fuel to vibrate, and to study their relation to reactor design features; to predict the vibratory response of a fuel assembly as a function of its design and environment, and; to study the relationship between vibration and fretting to determine when vibration results in fretting. (author)

Protecting AREVA ATRIUM™ BWR fuel from debris fretting failure

International Nuclear Information System (INIS)

Cole, Steven E.; Garner, Norman L.; Lippert, Hans-Joachim; Graebert, Rüdiger; Mollard, Pierre; Hahn, Gregory C.

2014-01-01

Historically, debris fretting has been the leading cause of fuel rod failure in BWR fuel assemblies, costing the industry millions of dollars in lost generation and negatively impacting the working area of plant site personnel. In this paper the focus will be on recent BWR fuel product innovation designed to eliminate debris related failures. Experience feedback from more than three decades of operation history with non-line-of-sight FUELGUARD™ lower tie plate debris filters will be presented. The development and relative effectiveness of successive generations of filtration technology will be discussed. It will be shown that modern, state of the art debris filters are an effective defense against debris fretting failure. Protective measures extend beyond inlet nozzle debris filters. The comprehensive debris resistance features built into AREVA’s newest fuel design, the ATRIUM™ 11, reduce the overall risk of debris entrapment as well as providing a degree of protection from debris that may fall down on the fuel assembly from above, e.g., during refueling operations. The positive recent experience in a debris sensitive plant will be discussed showing that the combination of advanced fuel technology and a robust foreign material exclusion program at the reactor site can eliminate the debris fretting failure mechanism. (author)

Sensitivity-Enhancement of FRET Immunoassays by Multiple-Antibody Conjugation on Quantum Dots.

Science.gov (United States)

Annio, Giacomo; Jennings, Travis; Tagit, Oya; Hildebrandt, Niko

2018-05-23

Quantum dots (QDs) are not only advantageous for color-tuning, improved brightness, and high stability, but their nanoparticle surfaces also allow for the attachment of many biomolecules. Because IgG antibodies (ABs) are in the same size range of biocompatible QDs and the AB orientation after conjugation to the QD is often random, it is difficult to predict if few or many ABs per QD will lead to an efficient AB-QD conjugate. This is particularly true for homogeneous Förster resonance energy transfer (FRET) sandwich immunoassays, for which the ABs on the QD must bind a biomarker that needs to bind a second AB-FRET-conjugate. Here, we investigate the performance of Tb-to-QD FRET immunoassays against total prostate specific antigen (TPSA) by changing the number of ABs per QD while leaving all the other assay components unchanged. We first characterize the AB-QD conjugation by various spectroscopic, microscopic, and chromatographic techniques and then quantify the TPSA immunoassay performance regarding sensitivity, limit of detection, and dynamic range. Our results show that an increasing conjugation ratio leads to significantly enhanced FRET immunoassays. These findings will be highly important for developing QD-based immunoassays in which the concentrations of both ABs and QDs can significantly influence the assay performance.

Extracting rate coefficients from single-molecule photon trajectories and FRET efficiency histograms for a fast-folding protein.

Science.gov (United States)

Chung, Hoi Sung; Gopich, Irina V; McHale, Kevin; Cellmer, Troy; Louis, John M; Eaton, William A

2011-04-28

Recently developed statistical methods by Gopich and Szabo were used to extract folding and unfolding rate coefficients from single-molecule Förster resonance energy transfer (FRET) data for proteins with kinetics too fast to measure waiting time distributions. Two types of experiments and two different analyses were performed. In one experiment bursts of photons were collected from donor and acceptor fluorophores attached to a 73-residue protein, α(3)D, freely diffusing through the illuminated volume of a confocal microscope system. In the second, the protein was immobilized by linkage to a surface, and photons were collected until one of the fluorophores bleached. Folding and unfolding rate coefficients and mean FRET efficiencies for the folded and unfolded subpopulations were obtained from a photon by photon analysis of the trajectories using a maximum likelihood method. The ability of the method to describe the data in terms of a two-state model was checked by recoloring the photon trajectories with the extracted parameters and comparing the calculated FRET efficiency histograms with the measured histograms. The sum of the rate coefficients for the two-state model agreed to within 30% with the relaxation rate obtained from the decay of the donor-acceptor cross-correlation function, confirming the high accuracy of the method. Interestingly, apparently reliable rate coefficients could be extracted using the maximum likelihood method, even at low (rate coefficients and mean FRET efficiencies were also obtained in an approximate procedure by simply fitting the FRET efficiency histograms, calculated by binning the donor and acceptor photons, with a sum of three-Gaussian functions. The kinetics are exposed in these histograms by the growth of a FRET efficiency peak at values intermediate between the folded and unfolded peaks as the bin size increases, a phenomenon with similarities to NMR exchange broadening. When comparable populations of folded and unfolded

Fluorescence resonance energy transfer (FRET) in chemistry and ...

Indian Academy of Sciences (India)

Förster distance dependence of the FRET rate. SANGEETA SAINI,1 HARJINDER SINGH2 and BIMAN BAGCHI1,*. 1Solid State and Structural Chemistry Unit, Indian Institute of Science, Bangalore 560 012. 2Permanent address: Department of ...

A combined wear-fatigue design methodology for fretting in the pressure armour layer of flexible marine risers

OpenAIRE

O'Halloran, S.M.; Shipway, P.H.; Connaire, A.D.; Leen, Sean B.; Harte, A.M.

2017-01-01

This paper presents a combined experimental and computational methodology for fretting wear-fatigue prediction of pressure armour wire in flexible marine risers. Fretting wear, friction and fatigue parameters of pressure armour material have been characterised experimentally. A combined fretting wear-fatigue finite element model has been developed using an adaptive meshing technique and the effect of bending-induced tangential slip has been characterised. It has been shown that a surface dama...

A dansyl-rhodamine chemosensor for Fe(III) based on off-on FRET.

Science.gov (United States)

Piao, Jingyu; Lv, Jia; Zhou, Xin; Zhao, Tong; Wu, Xue

2014-07-15

A novel fluorescent chemosensor bearing a rhodamine and a dansyl moiety was developed for highly selective detection of Fe(3+) based on fluorescence resonance energy transfer (FRET) mechanism. Binding of Fe(3+) to the chemosensor induced spirolactam ring opening in the rhodamine moiety and subsequent off-on FRET from the dansyl energy donor to the rhodamine energy acceptor due to the spectral overlap between the emission of the dansyl moiety and the absorption of the ring opened rhodamine moiety. Job's plot analysis indicated a 1:1 binding stoichiometry between the chemosensor and Fe(3+). The association constant was estimated to be 2.72×10(3) M(-1) according to the Benesi-Hildebrand method. With the feature of easy synthesis, simple structural skeleton and excellent sensing ability, the newly synthesized chemosensor provided the potential for applying as a highly selective fluorescent probe in complex samples containing various competitive metal ions and developing other metal ion chemosensors to fulfill various needs of biological and environmental field. Copyright © 2014 Elsevier B.V. All rights reserved.

Fretting fatigue behavior of high-strength steel monostrands under bending load

DEFF Research Database (Denmark)

Winkler, Jan; Georgakis, Christos T.; Fischer, Gregor

2015-01-01

In this paper, the fretting fatigue behavior of pretensioned high-strength steel monostrands is investigated. To measure the local deformations on the strands, a novel method based on the digital image correlation (DIC) technique was used to quantify the relative movement between individual wires...... along the length of the monostrand. Information about the monostrand bending stiffness and the extent of relative displacement between core and outer wires of a monostrand undergoing flexural deformations is provided. From the series of dynamic fatigue tests, a fretting fatigue spectrum is derived...

Improving brightness and photostability of green and red fluorescent proteins for live cell imaging and FRET reporting.

Science.gov (United States)

Bajar, Bryce T; Wang, Emily S; Lam, Amy J; Kim, Bongjae B; Jacobs, Conor L; Howe, Elizabeth S; Davidson, Michael W; Lin, Michael Z; Chu, Jun

2016-02-16

Many genetically encoded biosensors use Förster resonance energy transfer (FRET) to dynamically report biomolecular activities. While pairs of cyan and yellow fluorescent proteins (FPs) are most commonly used as FRET partner fluorophores, respectively, green and red FPs offer distinct advantages for FRET, such as greater spectral separation, less phototoxicity, and lower autofluorescence. We previously developed the green-red FRET pair Clover and mRuby2, which improves responsiveness in intramolecular FRET reporters with different designs. Here we report the engineering of brighter and more photostable variants, mClover3 and mRuby3. mClover3 improves photostability by 60% and mRuby3 by 200% over the previous generation of fluorophores. Notably, mRuby3 is also 35% brighter than mRuby2, making it both the brightest and most photostable monomeric red FP yet characterized. Furthermore, we developed a standardized methodology for assessing FP performance in mammalian cells as stand-alone markers and as FRET partners. We found that mClover3 or mRuby3 expression in mammalian cells provides the highest fluorescence signals of all jellyfish GFP or coral RFP derivatives, respectively. Finally, using mClover3 and mRuby3, we engineered an improved version of the CaMKIIα reporter Camuiα with a larger response amplitude.

Fluorescence resonance energy transfer (FRET-based subcellular visualization of pathogen-induced host receptor signaling

Directory of Open Access Journals (Sweden)

Zimmermann Timo

2009-11-01

Full Text Available Abstract Background Bacteria-triggered signaling events in infected host cells are key elements in shaping the host response to pathogens. Within the eukaryotic cell, signaling complexes are spatially organized. However, the investigation of protein-protein interactions triggered by bacterial infection in the cellular context is technically challenging. Here, we provide a methodological approach to exploit fluorescence resonance energy transfer (FRET to visualize pathogen-initiated signaling events in human cells. Results Live-cell microscopy revealed the transient recruitment of the Src family tyrosine kinase Hck upon bacterial engagement of the receptor carcinoembryonic antigen-related cell adhesion molecule 3 (CEACAM3. In cells expressing a CEACAM3 variant lacking the cytoplasmic domain, the Src homology 2 (SH2 domain of Hck (Hck-SH2 was not recruited, even though bacteria still bound to the receptor. FRET measurements on the basis of whole cell lysates revealed intimate binding between Hck-SH2 (using enhanced yellow fluorescent protein (YPet-Hck-SH2 and the tyrosine-phosphorylated enhanced cyan fluorescent protein-labeled cytoplasmic domain of wild-type CEACAM3 (CEACAM3 WT-CyPet and a flow cytometry-based FRET approach verified this association in intact cells. Using confocal microscopy and acceptor photobleaching, FRET between Hck-SH2 and CEACAM3 was localized to the sites of bacteria-host cell contact. Conclusion These data demonstrate not only the intimate binding of the SH2 domain of Hck to the tyrosine-phosphorylated cytoplasmic domain of CEACAM3 in intact cells, but furthermore, FRET measurements allow the subcellular localization of this process during bacterial infection. FRET-based assays are valuable tools to resolve bacteria-induced protein-protein interactions in the context of the intact host cell.

In Situ Probing Intracellular Drug Release from Redox-Responsive Micelles by United FRET and AIE.

Science.gov (United States)

Wang, Xuelin; Li, Juanjuan; Yan, Qi; Chen, Yanrui; Fan, Aiping; Wang, Zheng; Zhao, Yanjun

2018-03-01

Redox-responsive micelles are versatile nanoplatforms for on-demand drug delivery, but the in situ evaluation of drug release is challenging. Fluorescence resonance energy transfer (FRET) technique shows potential for addressing this, while the aggregation-caused quenching effect limits the assay sensitivity. The aim of the current work is to combine aggregation-induced emission (AIE) probe with FRET to realize drug release assessment from micelles. Tetraphenylethene (TPE) is selected as AIE dye and curcumin (Cur) is chosen as the model drug as well as FRET receptor. The drug is covalently linked to a block copolymer via the disulfide bond linker and TPE is also chemically linked to the polymer via an amide bond; the obtained amphiphilic polymer conjugate self-assembles into micelles with a hydrodynamic size of ≈125 nm. Upon the supplement of glutathione or tris(2-carboxyethyl)phosphine) trigger (10 × 10 -3 m), the drug release induces the fluorescence increase of both TPE and Cur. Accompanied with the FRET decay, absorption enhancement and particle size increase are observed. The same phenomenon is observed in MCF-7 cells. The FRET-AIE approach can be a useful addition to the spectrum of available methods for monitoring drug release from stimuli-responsive nanomedicine. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Advanced KSNP fuel, plus7 : grid-to-rod fretting wear resistance of the plus7 spacer grids

International Nuclear Information System (INIS)

Kim, Kyu Tae; Kim, Yong Hwan; Jang, Young Ki; Choi, Joon Hyung

2003-01-01

Vibration-induced grid-to-rod fretting wear initiates at a certain critical gap correlated with a critical work rate. A critical gap between grid and rod forms due to in-reactor performance of fuel, thermal relaxation of grid spring and irradiation growth of grid strap, etc. A critical work rate may be generated by three vibration mechanisms proposed in this paper. Three vibration mechanisms have been derived with various fretting wear experience in commercial reactors as well as various out-of-pile hydraulic test results. The first active vibration mechanism is high turbulence-induced excessive fuel rod vibration with the combination of excessive grid-to-rod gap. The second active vibration mechanism is self-excited fuel assembly vibration in a low frequency range caused by hydraulically unbalanced mixing vanes of the spacer grid assembly. The third active vibration mechanism is self-excited spacer grid strap vibration in quite a high frequency range caused by some spacer grid designs. In this study, each vibration mechanism on the grid-to-rod fretting wear damage is discussed. On the other hand, the effects of various grid designs on the fretting wear damage in the commercial reactors are predicted using the long-term fretting wear test results. It is found that the larger grid-to-rod initial contact area generates the less fretting wear damage. Consequently the conformal spring of PLUS7 is superior to typical convex shaped spring with regard to fretting wear resistance since the former generates relatively larger contact area than the latter

A Novel Water-soluble Ratiometric Fluorescent Probe Based on FRET for Sensing Lysosomal pH.

Science.gov (United States)

Song, Guang-Jie; Bai, Su-Yun; Luo, Jing; Cao, Xiao-Qun; Zhao, Bao-Xiang

2016-11-01

A new ratiometric fluorescent probe based on Förster resonance energy transfer (FRET) for sensing lysosomal pH has been developed. The probe (RMPM) was composed of imidazo[1,5-α]pyridine quaternary ammonium salt fluorophore as the FRET donor and the rhodamine moiety as the FRET acceptor. It's the first time to report that imidazo[1,5-α]pyridine quaternary ammonium salt acts as the FRET donor. The ratio of fluorescence intensity of the probe at two wavelengths (I 424 /I 581 ) changed significantly and responded linearly toward minor pH changes in the range of 5.4-6.6. It should be noted that it's rare to report that a ratiometric pH probe could detect so weak acidic pH with pKa = 6.31. In addition, probe RMPM exhibited excellent water-solubility, fast-response, all-right selectivity and brilliant reversibility. Moreover, RMPM has been successfully applied to sensing lysosomal pH in HeLa cells and has low cytotoxicity.

The role of FRET in solar concentrator efficiency and color tunability

Energy Technology Data Exchange (ETDEWEB)

Balaban, Benjamin, E-mail: bbalaban@ucsc.edu; Doshay, Sage; Osborn, Melissa; Rodriguez, Yvonne; Carter, Sue A., E-mail: sacarter@ucsc.edu

2014-02-15

We demonstrate concentration-dependent Förster-type energy transfer in a luminescent solar concentrator (LSC) material containing two high quantum yield laser dyes in a PMMA matrix. FRET heterotransfer is shown to be approximately 50% efficient in the regime of 2×10{sup −3}molal acceptor dye by weight in the host polymer. The two dyes used have been well studied for solar concentrator applications: BASF's Lumogen Red 305, and Exciton Chemical Company's DCM both demonstrate desirable stability, quantum yield, and complementary absorption spectra. We demonstrate how multiple-dye LSC devices employing FRET increase the absorption of air mass 1.5 solar irradiance without affecting the self-absorption properties of the film. Color tunability may be achieved through the addition of additional absorbers while minimizing the impact on waveguide efficiency. -- Highlights: • Förster Resonance Energy Transfer is demonstrated in a two-dye luminescent solar concentrator. • Donor-acceptor pair distance is related to the dye concentration in PMMA. • FRET's benefit to waveguide transport losses and color tunability is discussed.

Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization

Science.gov (United States)

Okamura, Yukio; Kondo, Satoshi; Sase, Ichiro; Suga, Takayuki; Mise, Kazuyuki; Furusawa, Iwao; Kawakami, Shigeki; Watanabe, Yuichiro

2000-01-01

A set of fluorescently-labeled DNA probes that hybridize with the target RNA and produce fluorescence resonance energy transfer (FRET) signals can be utilized for the detection of specific RNA. We have developed probe sets to detect and discriminate single-strand RNA molecules of plant viral genome, and sought a method to improve the FRET signals to handle in vivo applications. Consequently, we found that a double-labeled donor probe labeled with Bodipy dye yielded a remarkable increase in fluorescence intensity compared to a single-labeled donor probe used in an ordinary FRET. This double-labeled donor system can be easily applied to improve various FRET probes since the dependence upon sequence and label position in enhancement is not as strict. Furthermore this method could be applied to other nucleic acid substances, such as oligo RNA and phosphorothioate oligonucleotides (S-oligos) to enhance FRET signal. Although the double-labeled donor probes labeled with a variety of fluorophores had unexpected properties (strange UV-visible absorption spectra, decrease of intensity and decay of donor fluorescence) compared with single-labeled ones, they had no relation to FRET enhancement. This signal amplification mechanism cannot be explained simply based on our current results and knowledge of FRET. Yet it is possible to utilize this double-labeled donor system in various applications of FRET as a simple signal-enhancement method. PMID:11121494

QUANTITATIVE CONFOCAL LASER SCANNING MICROSCOPY

Directory of Open Access Journals (Sweden)

Merete Krog Raarup

2011-05-01

Full Text Available This paper discusses recent advances in confocal laser scanning microscopy (CLSM for imaging of 3D structure as well as quantitative characterization of biomolecular interactions and diffusion behaviour by means of one- and two-photon excitation. The use of CLSM for improved stereological length estimation in thick (up to 0.5 mm tissue is proposed. The techniques of FRET (Fluorescence Resonance Energy Transfer, FLIM (Fluorescence Lifetime Imaging Microscopy, FCS (Fluorescence Correlation Spectroscopy and FRAP (Fluorescence Recovery After Photobleaching are introduced and their applicability for quantitative imaging of biomolecular (co-localization and trafficking in live cells described. The advantage of two-photon versus one-photon excitation in relation to these techniques is discussed.

FRET enhancement close to gold nanoparticles positioned in DNA origami constructs.

Science.gov (United States)

Aissaoui, Nesrine; Moth-Poulsen, Kasper; Käll, Mikael; Johansson, Peter; Wilhelmsson, L Marcus; Albinsson, Bo

2017-01-05

Here we investigate the energy transfer rates of a Förster resonance energy transfer (FRET) pair positioned in close proximity to a 5 nm gold nanoparticle (AuNP) on a DNA origami construct. We study the distance dependence of the FRET rate by varying the location of the donor molecule, D, relative to the AuNP while maintaining a fixed location of the acceptor molecule, A. The presence of the AuNP induces an alteration in the spontaneous emission of the donor (including radiative and non-radiative rates) which is strongly dependent on the distance between the donor and AuNP surface. Simultaneously, the energy transfer rates are enhanced at shorter D-A (and D-AuNP) distances. Overall, in addition to the direct influence of the acceptor and AuNP on the donor decay there is also a significant increase in decay rate not explained by the sum of the two interactions. This leads to enhanced energy transfer between donor and acceptor in the presence of a 5 nm AuNP. We also demonstrate that the transfer rate in the three "particle" geometry (D + A + AuNP) depends approximately linearly on the transfer rate in the donor-AuNP system, suggesting the possibility to control FRET process with electric field induced by 5 nm AuNPs close to the donor fluorophore. It is concluded that DNA origami is a very versatile platform for studying interactions between molecules and plasmonic nanoparticles in general and FRET enhancement in particular.

Polyfluorophore Excimers and Exciplexes as FRET Donors in DNA

Science.gov (United States)

Teo, Yin Nah; Kool, Eric T.

2009-01-01

We describe studies aimed at testing whether oligomeric exciplex- and excimer fluorophores conjugated to DNA have the potential to act as donors for energy transfer by the Förster mechanism. Oligodeoxyfluorosides (ODFs) are composed of stacked, electronically interacting fluorophores replacing the bases on a DNA scaffold. The monomer chromophores in the twenty tetramer-length ODFs studied here include pyrene (Y), benzopyrene (B), perylene (E), dimethylaminostilbene (D), and a nonfluorescent spacer (S); these are conjugated in varied combinations at the 3’ end of a 14mer DNA probe sequence. In the absence of an acceptor chromophore, many of the ODF-DNAs show broad, unstructured long-wavelength emission peaks characteristic of excimer and exciplex excited states, similar to what has been observed for unconjugated ODFs. Although such delocalized excited states have been widely studied, we know of no prior report of their use in FRET. We tested the ability of the twenty ODFs to donate energy to Cy5 and TAMRA dyes conjugated to a complementary strand of DNA, with these acceptors oriented either at the near or far end of the ODF-conjugated probes. Results showed that a number of the ODF fluorophores exhibited relatively efficient energy transfer characteristic of the Förster mechanism, as judged by drops in donor emission quantum yield and fluorescence lifetime, accompanied by increases in intensity of acceptor emission bands. Excimer/exciplex bands in the donors were selectively quenched while shorter-wavelength monomer emission stayed relatively constant, consistent with the notion that the delocalized excited states, rather than individual fluorophores, are the donors. Interestingly, only specific sequences of ODFs were able to act as donors, while others did not, even though their emission wavelengths were similar. The new FRET donors possess large Stokes shifts, which can be beneficial for multiple applications. In addition, all ODFs can be excited at a single

Development of the vibration analysis technique of fuel rod and research on the methodology of fuel fretting wear analysis

International Nuclear Information System (INIS)

Kang, Heung Seok; Kim, Kyung Kyu; Yoon, Hyung Hoo; Song, Ki Nam

1998-12-01

The FEM program has been developed to predict the natural frequencies, the FEM program has been developed to predict the natural frequencies, and mode shapes of fuel rod subjected to axial force and continuously supported by a rotational and vent spring system, and to calculate the minimum reaction forces of the spacer grid spring when the maximum vibration amplitude of fuel rod is known. This program has been verified by commercial ANSYS program and the vibration test of dummy rods in air. The test equipment were set to get the fifth modes of test rods. Partial slip problem has been studied for the analysis of fuel fretting problem. Firstly, the assumption of semi-infiniteness of the contact bodies were validated by finite element (FE) analysis. From FE results, a classical bodies were validated by finite element (FE) analysis. From FE results, a classical theory of elasticity was utilized with regarding the problem as a plane problem. Secondly, the Mindlin-Cattaneo problem was re-evaluated, which gave the fundamental idea for developing the numerical tool for the shear traction on the contact. Shear force of sequentially-changing directions was considered and the corresponding shear traction was evaluated by extending the numerical tool for the Mindlin-Cattaneo problem

Development of the vibration analysis technique of fuel rod and research on the methodology of fuel fretting wear analysis

Energy Technology Data Exchange (ETDEWEB)

Kang, Heung Seok; Kim, Kyung Kyu; Yoon, Hyung Hoo; Song, Ki Nam

1998-12-01

The FEM program has been developed to predict the natural frequencies, the FEM program has been developed to predict the natural frequencies, and mode shapes of fuel rod subjected to axial force and continuously supported by a rotational and vent spring system, and to calculate the minimum reaction forces of the spacer grid spring when the maximum vibration amplitude of fuel rod is known. This program has been verified by commercial ANSYS program and the vibration test of dummy rods in air. The test equipment were set to get the fifth modes of test rods. Partial slip problem has been studied for the analysis of fuel fretting problem. Firstly, the assumption of semi-infiniteness of the contact bodies were validated by finite element (FE) analysis. From FE results, a classical bodies were validated by finite element (FE) analysis. From FE results, aclassical theory of elasticity was utilized with regarding the problem as a plane problem. Secondly, the Mindlin-Cattaneo problem was re-evaluated, which gave the fundamental idea for developing the numerical tool for the shear traction on the contact. Shear force of sequentially-changing directions was considered and the corresponding shear traction was evaluated by extending the numerical tool for the Mindlin-Cattaneo problem.

Nucleic Acid Base Analog FRET-Pair Facilitating Detailed Structural Measurements in Nucleic Acid Containing Systems

DEFF Research Database (Denmark)

Börjesson, Karl; Preus, Søren; El-Sagheer, Afaf

2009-01-01

We present the first nucleobase analog fluorescence resonance energy transfer (FRET)-pair. The pair consists of tCO, 1,3-diaza-2-oxophenoxazine, as an energy donor and the newly developed tC(nitro), 7-nitro-1,3-diaza-2-oxophenothiazine, as an energy acceptor. The FRET-pair successfully monitors d...

Burst pressure and leak rate from fretted SG tubes

International Nuclear Information System (INIS)

Hwang, Seong Sik; Jung, Man Kyo; Kim, Hong Pyo; Kim, Joung Soo

2005-01-01

Steam generator(SG) tubes of a pressurized water reactor(PWR) have suffered from various types of corrosion, such as pitting, wastage and stress corrosion cracking (SCC) on both the primary and secondary side. Recently, fretting/wear degradation at the tube support region has been reported in some Korean nuclear power plants. In order to prevent the primary coolant from leaking to the secondary side, the tubes are repaired by a sleeving or plugging. It is important to establish the repair criteria to assure a reactor integrity and yet maintain the plugging ratio within the limits needed for an efficient operation. The objective of the burst test is to obtain a relationship between the burst/leak rate and the shape of the fretted flaws machined with an electro discharge machining (EDM)

Conformational Analysis of Misfolded Protein Aggregation by FRET and Live-Cell Imaging Techniques

Directory of Open Access Journals (Sweden)

Akira Kitamura

2015-03-01

Full Text Available Cellular homeostasis is maintained by several types of protein machinery, including molecular chaperones and proteolysis systems. Dysregulation of the proteome disrupts homeostasis in cells, tissues, and the organism as a whole, and has been hypothesized to cause neurodegenerative disorders, including amyotrophic lateral sclerosis (ALS and Huntington’s disease (HD. A hallmark of neurodegenerative disorders is formation of ubiquitin-positive inclusion bodies in neurons, suggesting that the aggregation process of misfolded proteins changes during disease progression. Hence, high-throughput determination of soluble oligomers during the aggregation process, as well as the conformation of sequestered proteins in inclusion bodies, is essential for elucidation of physiological regulation mechanism and drug discovery in this field. To elucidate the interaction, accumulation, and conformation of aggregation-prone proteins, in situ spectroscopic imaging techniques, such as Förster/fluorescence resonance energy transfer (FRET, fluorescence correlation spectroscopy (FCS, and bimolecular fluorescence complementation (BiFC have been employed. Here, we summarize recent reports in which these techniques were applied to the analysis of aggregation-prone proteins (in particular their dimerization, interactions, and conformational changes, and describe several fluorescent indicators used for real-time observation of physiological states related to proteostasis.

Quantitative investment analysis

CERN Document Server

DeFusco, Richard

2007-01-01

In the "Second Edition" of "Quantitative Investment Analysis," financial experts Richard DeFusco, Dennis McLeavey, Jerald Pinto, and David Runkle outline the tools and techniques needed to understand and apply quantitative methods to today's investment process.

Ultrasensitive FRET-based DNA sensor using PNA/DNA hybridization.

Science.gov (United States)

Yang, Lan-Hee; Ahn, Dong June; Koo, Eunhae

2016-12-01

In the diagnosis of genetic diseases, rapid and highly sensitive DNA detection is crucial. Therefore, many strategies for detecting target DNA have been developed, including electrical, optical, and mechanical methods. Herein, a highly sensitive FRET based sensor was developed by using PNA (Peptide Nucleic Acid) probe and QD, in which red color QDs are hybridized with capture probes, reporter probes and target DNAs by EDC-NHS coupling. The hybridized probe with target DNA gives off fluorescent signal due to the energy transfer from QD to Cy5 dye in the reporter probe. Compared to the conventional DNA sensor using DNA probes, the DNA sensor using PNA probes shows higher FRET factor and efficiency due to the higher reactivity between PNA and target DNA. In addition, to elicit the effect of the distance between the donor and the acceptor, we have investigated two types of the reporter probes having Cy5 dyes attached at the different positions of the reporter probes. Results show that the shorter the distance between QDs and Cy5s, the stronger the signal intensity. Furthermore, based on the fluorescence microscopy images using microcapillary chips, the FRET signal is enhanced to be up to 276% times stronger than the signal obtained using the cuvette by the fluorescence spectrometer. These results suggest that the PNA probe system conjugated with QDs can be used as ultrasensitive DNA nanosensors. Copyright © 2016. Published by Elsevier B.V.

Simulation of vibration modes of the fuel rod damaged due to the grid-to-rod fretting wear

International Nuclear Information System (INIS)

Kim, Kyu Tae; Kim, Kyeong Koo; Jang, Young Ki; Lee, Kyou Seok

1997-01-01

The flow-induced fuel fretting wear observed in some PWRs mainly proceeds in the grid-to-rod contact positions. The grid-to-rod fretting wear in the PWR fuel assembly depends on grid-to-rod gap size, its axial profile and flow-induced vibration. This paper describes the GRIDFORCE program which generates the axially dependent grid-to-rod gap size as a function of burnup. The axially dependent grid-to-rod gap profiles are employed to predict the fuel rod vibration mode shapes by the ANSYS code. With the help of the Paidousis empirical formula, this paper also calculates the fuel rod vibration amplitudes under various supporting conditions, which indicates that the increase of the number of unsupported mid-grids will increase the fuel rod vibration amplitude. On the other hand, the comparison of the predicted vibration mode shapes and the observed mid-grid fretting wear pattern indicates that the 1st and 6th vibration mode shapes under the supporting inactive condition at the mid-grids can simulate the observed mid-grid fretting wear profile. This paper also proposes design guidelines against the grid-to-rod fretting wear. (author). 3 refs., 8 figs

Fretting Corrosion Behavior of Experimental Ti-20Cr Compared to Titanium.

Science.gov (United States)

Sawada, Tomofumi; Schille, Christine; Almadani, Atif; Geis-Gerstorfer, Jürgen

2017-02-17

Experimental cast titanium alloys containing 20 mass% chromium (Ti-20Cr) show preferable mechanical properties and a good corrosion resistance. This study evaluated the fretting corrosion behavior of Ti-20Cr. Ti-20Cr ( n = 4) and commercially pure titanium (CP-Ti, n = 6) disk specimens were used. The fretting corrosion test was performed by electrochemical corrosion at 0.3 V in 0.9% saline solution and mechanical damage using 10 scratching cycles with three different scratching speeds (10-40 mm/s) at 10 N. After testing, the activation peak, repassivation time and surface morphology of each specimen were analyzed. The differences between the results were tested by parametric tests (α = 0.05). The average activation peaks were significantly higher in CP-Ti than in Ti-20Cr ( p Ti. Slight differences in the repassivation time were observed between the materials at every scratching speed; faster scratching speeds showed shorter repassivation times in both materials ( p Ti showed severe damage and significantly higher wear depth than Ti-20Cr ( p < 0.05). In conclusion, adding chromium to titanium reduced surface damage and improved the fretting corrosion resistance.

Fluorescent protein pair emit intracellular FRET signal suitable for FACS screening

International Nuclear Information System (INIS)

Johansson, Daniel X.; Brismar, Hjalmar; Persson, Mats A.A.

2007-01-01

The fluorescent proteins ECFP and HcRed were shown to give an easily resolved FRET-signal when expressed as a fusion inside mammalian cells. HeLa-tat cells expressing ECFP, pHcRed, or the fusion protein pHcRed-ECFP were analyzed by flow cytometry after excitation of ECFP. Cells expressing HcRed-ECFP, or ECFP and HcRed, were mixed and FACS-sorted for FRET positive cells: HcRed-ECFP cells were greatly enriched (72 times). Next, cloned human antibodies were fused with ECFP and expressed anchored to the ER membrane. Their cognate antigens (HIV-1 gp120 or gp41) were fused to HcRed and co-expressed in the ER. An increase of 13.5 ± 1.5% (mean ± SEM) and 8.0 ± 0.7% in ECFP fluorescence for the specific antibodies reacting with gp120 or gp41, respectively, was noted after photobleaching. A positive control (HcRed-ECFP) gave a 14.8 ± 2.6% increase. Surprisingly, the unspecific antibody (anti-TT) showed 12.1 ± 1.1% increase, possibly because overexpression in the limited ER compartment gave false FRET signals

Determination of a Wear Initiation Cycle by using a Contact Resistance Measurement in Nuclear Fuel Fretting

International Nuclear Information System (INIS)

Lee, Young Ho; Kim, Hyung Kyu

2008-01-01

In nuclear fuel fretting, the improving of the contact condition with a modified spring shape is a useful method for increasing the wear resistance of the nuclear fuel rod. This is because the fretting wear resistance between the fuel rod and grid spring is mainly affected by the grid spring shape rather than the environment, the contact modes, etc. In addition, the wear resistance is affected by the wear debris behavior between contact surfaces. So, it is expected that the wear initiation of each spring shape should be determined in order to evaluate a wear resistance. However, it is almost impossible to measure the wear behavior in contact surfaces on a real time basis because the contact surfaces are always hidden. Besides, the results of the worn surface observation after the fretting wear tests are restricted to archive the information on the wear debris behavior and the formation mechanism of the wear scar. In order to evaluate the wear behavior during the fretting wear tests, it is proposed that the contact resistance measurement is a useful method for examining the wear initiation cycle and modes. Generally, fretting wear damages are rapidly progressed by a localized plastic deformation between the contact surfaces, crack initiation and fracture of the deformed surface with a strain hardening difference between a surface and a subsurface and finally a detachment of wear debris. After this, wear debris is easily oxidized by frictional heat, test environment, etc. At this time, a small amount of electric current applied between the contact surfaces will be influenced by the wear debris, which could be an obstacle to an electric current flow. So, it is possible to archive the information on the wear behavior by measuring the contact resistance. In order to determine the wear initiation cycle during the fretting wear tests, in this study, fretting wear tests have been performed by applying a constant electric current in room temperature air

Fretting wear damage of steam generator tubes and its prediction modeling

International Nuclear Information System (INIS)

Che Honglong; Lei Mingkai

2013-01-01

The steam generator is the key equipment used for the energy transition in nuclear power plant. Since the high-temperature and high-pressure fluid flows with high speed, the steam generator tubes will be excited and vibrate, leading to the tremendous fretting wear problem on the tubes, sometimes even leading to tube cracking. This paper introduces typical fretting wear cases, the result of corresponding simulation wear experiment and damage mechanism which combining mechanical wear and erosion-corrosion. Work rate model could give a reasonable life prediction about the steam generator tube, and this predictive model has been used in nuclear power plant safety assessment. (authors)

Stem Migration and Fretting Corrosion of the Antirotation Pin in the K2/Apex Hip System.

Science.gov (United States)

Kent, Michael; Edmondson, Mark; Ebert, Jay; Nivbrant, Nils; Kop, Alan; Wood, David; De Steiger, Richard

2016-03-01

Many exchangeable neck hip systems have been withdrawn because of fretting corrosion at the neck/stem coupling. Our prospective randomized study evaluating stem stability (Roentgen stereophotogrammetric analysis, dual-energy x-ray absorptiometry) and clinical outcomes between the K2/Apex hip systems was ceased early because of a withdrawal of the stems which had an unfavorably high early revision rate reported in the Australian Orthopaedic Association National Joint Registry (9.3% at 3 years). At 2 years, there are no clinical differences between the stems. Roentgen stereophotogrammetric analysis has identified a high proportion of potentially concerning subsidence and retroversion in both groups, more marked in the K2 stem, although mostly in asymptomatic patients. Dual-energy x-ray absorptiometry has shown similar bone density around the stems. Retrieval analysis of 3 study patients showed fretting corrosion of the antirotation pin and aseptic lymphocyte-dominated vasculitis-associated lesion, with no relationship to bearing type or size. Analysis of 7 further nonstudy K2/Apex stems confirmed similar corrosion. This study shows potentially concerning subsidence of both stems and is the first to describe corrosion at the neck-stem interface and a relationship to metal-related pathology. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

Development of FRET biosensors for mammalian and plant systems

NARCIS (Netherlands)

Hamers, D.; van Voorst Vader, L.; Borst, J.W.; Goedhart, J.

2014-01-01

Genetically encoded biosensors are increasingly used in visualising signalling processes in different organisms. Sensors based on green fluorescent protein technology are providing a great opportunity for using Forster resonance energy transfer (FRET) as a tool that allows for monitoring dynamic

Studying DNA looping by single-molecule FRET.

Science.gov (United States)

Le, Tung T; Kim, Harold D

2014-06-28

Bending of double-stranded DNA (dsDNA) is associated with many important biological processes such as DNA-protein recognition and DNA packaging into nucleosomes. Thermodynamics of dsDNA bending has been studied by a method called cyclization which relies on DNA ligase to covalently join short sticky ends of a dsDNA. However, ligation efficiency can be affected by many factors that are not related to dsDNA looping such as the DNA structure surrounding the joined sticky ends, and ligase can also affect the apparent looping rate through mechanisms such as nonspecific binding. Here, we show how to measure dsDNA looping kinetics without ligase by detecting transient DNA loop formation by FRET (Fluorescence Resonance Energy Transfer). dsDNA molecules are constructed using a simple PCR-based protocol with a FRET pair and a biotin linker. The looping probability density known as the J factor is extracted from the looping rate and the annealing rate between two disconnected sticky ends. By testing two dsDNAs with different intrinsic curvatures, we show that the J factor is sensitive to the intrinsic shape of the dsDNA.

Precision and accuracy in smFRET based structural studies—A benchmark study of the Fast-Nano-Positioning System

Science.gov (United States)

Nagy, Julia; Eilert, Tobias; Michaelis, Jens

2018-03-01

Modern hybrid structural analysis methods have opened new possibilities to analyze and resolve flexible protein complexes where conventional crystallographic methods have reached their limits. Here, the Fast-Nano-Positioning System (Fast-NPS), a Bayesian parameter estimation-based analysis method and software, is an interesting method since it allows for the localization of unknown fluorescent dye molecules attached to macromolecular complexes based on single-molecule Förster resonance energy transfer (smFRET) measurements. However, the precision, accuracy, and reliability of structural models derived from results based on such complex calculation schemes are oftentimes difficult to evaluate. Therefore, we present two proof-of-principle benchmark studies where we use smFRET data to localize supposedly unknown positions on a DNA as well as on a protein-nucleic acid complex. Since we use complexes where structural information is available, we can compare Fast-NPS localization to the existing structural data. In particular, we compare different dye models and discuss how both accuracy and precision can be optimized.

Simple estimation of Förster Resonance Energy Transfer (FRET) orientation factor distribution in membranes.

Science.gov (United States)

Loura, Luís M S

2012-11-19

Because of its acute sensitivity to distance in the nanometer scale, Förster resonance energy transfer (FRET) has found a large variety of applications in many fields of chemistry, physics, and biology. One important issue regarding the correct usage of FRET is its dependence on the donor-acceptor relative orientation, expressed as the orientation factor k(2). Different donor/acceptor conformations can lead to k(2) values in the 0 ≤ k(2) ≤ 4 range. Because the characteristic distance for FRET, R(0), is proportional to (k(2))1/6, uncertainties in the orientation factor are reflected in the quality of information that can be retrieved from a FRET experiment. In most cases, the average value of k(2) corresponding to the dynamic isotropic limit ( = 2/3) is used for computation of R(0) and hence donor-acceptor distances and acceptor concentrations. However, this can lead to significant error in unfavorable cases. This issue is more critical in membrane systems, because of their intrinsically anisotropic nature and their reduced fluidity in comparison to most common solvents. Here, a simple numerical simulation method for estimation of the probability density function of k(2) for membrane-embedded donor and acceptor fluorophores in the dynamic regime is presented. In the simplest form, the proposed procedure uses as input the most probable orientations of the donor and acceptor transition dipoles, obtained by experimental (including linear dichroism) or theoretical (such as molecular dynamics simulation) techniques. Optionally, information about the widths of the donor and/or acceptor angular distributions may be incorporated. The methodology is illustrated for special limiting cases and common membrane FRET pairs.

Effect of different atmospheres on the electrical contact performance of electronic components under fretting wear

Science.gov (United States)

Liu, Xin-Long; Cai, Zhen-Bing; Cui, Ye; Liu, Shan-Bang; Xu, Xiao-Jun; Zhu, Min-Hao

2018-04-01

The effects of oxide etch on the surface morphology of metals for industrial application is a common cause of electrical contacts failure, and it has becomes a more severe problem with the miniaturization of modern electronic devices. This study investigated the effects of electrical contact resistance on the contactor under three different atmospheres (oxygen, air, and nitrogen) based on 99.9% copper/pogo pins contacts through fretting experiments. The results showed the minimum and stable electrical contact resistance value when shrouded in the nitrogen environment and with high friction coefficient. The rich oxygen environment promotes the formation of cuprous oxide, thereby the electrical contact resistance increases. Scanning electron microscope microscopy and electron probe microanalysis were used to analyze the morphology and distribution of elements of the wear area, respectively. The surface product between contacts was investigated by x-ray photoelectron spectroscopy analysis to explain the different electrical contact properties of the three tested samples during fretting.

Graphene and graphene-like two-denominational materials based fluorescence resonance energy transfer (FRET) assays for biological applications.

Science.gov (United States)

Tian, Feng; Lyu, Jing; Shi, Jingyu; Yang, Mo

2017-03-15

In the past decades, Förster resonance energy transfer (FRET) has been applied in many biological applications to reveal the biological information at the nanoscale. Recently, graphene and graphene-like two-dimensional (2D) nanomaterials started to be used in FRET assays as donors or acceptors including graphene oxide (GO), graphene quantum dot (GQD), graphitic-carbon nitride nanosheets (g-C 3 N 4 ) and transition metal dichalcogenides (e.g. MoS 2 , MnO 2, and WS 2 ). Due to the remarkable properties such as large surface to volume ratio, tunable energy band, photoluminescence and excellent biocompatibility, these 2D nanomaterials based FRET assays have shown great potential in various biological applications. This review summarizes the recent development of graphene and graphene-like 2D nanomaterials based FRET assays in applications of biosensing, bioimaging, and drug delivery monitoring. Copyright © 2016 Elsevier B.V. All rights reserved.

Real-time determination of intracellular oxygen in bacteria using a genetically encoded FRET-based biosensor

Directory of Open Access Journals (Sweden)

Potzkei Janko

2012-03-01

Full Text Available Abstract Background Molecular oxygen (O2 is one of the key metabolites of all obligate and facultative aerobic pro- and eukaryotes. It plays a fundamental role in energy homeostasis whereas oxygen deprivation, in turn, broadly affects various physiological and pathophysiological processes. Therefore, real-time monitoring of cellular oxygen levels is basically a prerequisite for the analysis of hypoxia-induced processes in living cells and tissues. Results We developed a genetically encoded Förster resonance energy transfer (FRET-based biosensor allowing the observation of changing molecular oxygen concentrations inside living cells. This biosensor named FluBO (fluorescent protein-based biosensor for oxygen consists of the yellow fluorescent protein (YFP that is sensitive towards oxygen depletion and the hypoxia-tolerant flavin-binding fluorescent protein (FbFP. Since O2 is essential for the formation of the YFP chromophore, efficient FRET from the FbFP donor domain to the YFP acceptor domain only occurs in the presence but not in the absence of oxygen. The oxygen biosensor was used for continuous real-time monitoring of temporal changes of O2 levels in the cytoplasm of Escherichia coli cells during batch cultivation. Conclusions FluBO represents a unique FRET-based oxygen biosensor which allows the non-invasive ratiometric readout of cellular oxygen. Thus, FluBO can serve as a novel and powerful probe for investigating the occurrence of hypoxia and its effects on a variety of (pathophysiological processes in living cells.

Developing Fast Fluorescent Protein Voltage Sensors by Optimizing FRET Interactions.

Directory of Open Access Journals (Sweden)

Uhna Sung

Full Text Available FRET (Förster Resonance Energy Transfer-based protein voltage sensors can be useful for monitoring neuronal activity in vivo because the ratio of signals between the donor and acceptor pair reduces common sources of noise such as heart beat artifacts. We improved the performance of FRET based genetically encoded Fluorescent Protein (FP voltage sensors by optimizing the location of donor and acceptor FPs flanking the voltage sensitive domain of the Ciona intestinalis voltage sensitive phosphatase. First, we created 39 different "Nabi1" constructs by positioning the donor FP, UKG, at 8 different locations downstream of the voltage-sensing domain and the acceptor FP, mKO, at 6 positions upstream. Several of these combinations resulted in large voltage dependent signals and relatively fast kinetics. Nabi1 probes responded with signal size up to 11% ΔF/F for a 100 mV depolarization and fast response time constants both for signal activation (~2 ms and signal decay (~3 ms. We improved expression in neuronal cells by replacing the mKO and UKG FRET pair with Clover (donor FP and mRuby2 (acceptor FP to create Nabi2 probes. Nabi2 probes also had large signals and relatively fast time constants in HEK293 cells. In primary neuronal culture, a Nabi2 probe was able to differentiate individual action potentials at 45 Hz.

Experimental evaluation of the fretting fatigue behavior of high-strength steel monostrands

DEFF Research Database (Denmark)

Winkler, Jan; Fischer, Gregor; Georgakis, Christos T.

2013-01-01

In this paper, the fretting fatigue behavior of pretensioned high-strength steel monostrands is investigated. A method based on the digital image correlation (DIC) technique was used to quantify the relative movement between individual wires along the length of the monostrand. The experimental data....... Moreover, the paper provides relevant information about the monostrand bending stiffness and the extent of relative displacement between core and outer wires of the monostrand undergoing flexural deformations. The results presented herein are of special interest for the fatigue analysis of modern stay...

Spectroscopic and theoretical investigation of conformational changes of proteins by synthesized pyrimidine derivative and its sensitivity towards FRET application

Science.gov (United States)

Ghosh, Swadesh; Singharoy, Dipti; Bhattacharya, Subhash Chandra

2018-04-01

Interest in synthesizing and characterizing (IR, NMR and HRMS spectroscopic methods) a pyrimidine based Schiff-base ligand, 2-(2-(Anthracen-9-ylmethylene) hydrazinyl)-4,6-dimethyl pyrimidine (ANHP) has been developed for its application to ascertain the conformational change of protein and sensitivity towards fluorescence resonance energy transfer (FRET) process. Location of ANHP in bovine serum albumin (BSA) and human serum albumin (HSA) proteins environment has been determined using different spectroscopic techniques. Weakly fluorescent ANHP have shown greater protein induced fluorescence enhancement (PIFE) in case of HSA than BSA, though in both cases energy transfer efficiency are almost same but difference in binding constant values encourages us to find the location of ANHP within the complex protein environment. From the FRET parameter and α-helicity change, it has been found that ANHP bound with Trp-214 of HSA and surface Trp-134 of BSA. Conformational changes of proteins have been observed more for HSA than BSA in presence of ANHP, which has confirmed the location of ANHP in both the protein environments. Coupled with experimental studies, molecular docking analysis has also been done to explain the locations and distance dependent FRET process of ANHP in both proteins.

Association of Myosin Va and Schwann cells-derived RNA in mammal myelinated axons, analyzed by immunocytochemistry and confocal FRET microscopy.

Science.gov (United States)

Canclini, Lucía; Wallrabe, Horst; Di Paolo, Andrés; Kun, Alejandra; Calliari, Aldo; Sotelo-Silveira, José Roberto; Sotelo, José Roberto

2014-03-15

Evidence from multiple sources supports the hypothesis that Schwann cells in the peripheral nervous system transfer messenger RNA and ribosomes to the axons they ensheath. Several technical and methodological difficulties exist for investigators to unravel this process in myelinated axons - a complex two-cell unit. We present an experimental design to demonstrate that newly synthesized RNA is transferred from Schwann cells to axons in association with Myosin Va. The use of quantitative confocal FRET microscopy to track newly-synthesized RNA and determine the molecular association with Myosin Va, is described in detail. Copyright © 2013 Elsevier Inc. All rights reserved.

The spatiotemporal pattern of Src activation at lipid rafts revealed by diffusion-corrected FRET imaging.

Directory of Open Access Journals (Sweden)

Shaoying Lu

2008-07-01

Full Text Available Genetically encoded biosensors based on fluorescence resonance energy transfer (FRET have been widely applied to visualize the molecular activity in live cells with high spatiotemporal resolution. However, the rapid diffusion of biosensor proteins hinders a precise reconstruction of the actual molecular activation map. Based on fluorescence recovery after photobleaching (FRAP experiments, we have developed a finite element (FE method to analyze, simulate, and subtract the diffusion effect of mobile biosensors. This method has been applied to analyze the mobility of Src FRET biosensors engineered to reside at different subcompartments in live cells. The results indicate that the Src biosensor located in the cytoplasm moves 4-8 folds faster (0.93+/-0.06 microm(2/sec than those anchored on different compartments in plasma membrane (at lipid raft: 0.11+/-0.01 microm(2/sec and outside: 0.18+/-0.02 microm(2/sec. The mobility of biosensor at lipid rafts is slower than that outside of lipid rafts and is dominated by two-dimensional diffusion. When this diffusion effect was subtracted from the FRET ratio images, high Src activity at lipid rafts was observed at clustered regions proximal to the cell periphery, which remained relatively stationary upon epidermal growth factor (EGF stimulation. This result suggests that EGF induced a Src activation at lipid rafts with well-coordinated spatiotemporal patterns. Our FE-based method also provides an integrated platform of image analysis for studying molecular mobility and reconstructing the spatiotemporal activation maps of signaling molecules in live cells.

Branched DNA nanostructures efficiently stabilised and monitored by novel pyrene-perylene 2'-α-l-amino-LNA FRET pairs

DEFF Research Database (Denmark)

Astakhova, I Kira; Santhosh Kumar, T; Campbell, Meghan A

2013-01-01

Novel pyrene-perylene α-l-LNA FRET pairs described herein effectively detect assembly of 2- and 3-way branched DNA nanostructures prepared by postsynthetic microwave-assisted CuAAC click chemistry. The fluorescent signalling of assembly by internally positioned FRET pairs is achieved with low...

Effect of the achondroplasia mutation on FGFR3 dimerization and FGFR3 structural response to fgf1 and fgf2: A quantitative FRET study in osmotically derived plasma membrane vesicles.

Science.gov (United States)

Sarabipour, Sarvenaz; Hristova, Kalina

2016-07-01

The G380R mutation in the transmembrane domain of FGFR3 is a germline mutation responsible for most cases of Achondroplasia, a common form of human dwarfism. Here we use quantitative Fӧster Resonance Energy Transfer (FRET) and osmotically derived plasma membrane vesicles to study the effect of the achondroplasia mutation on the early stages of FGFR3 signaling in response to the ligands fgf1 and fgf2. Using a methodology that allows us to capture structural changes on the cytoplasmic side of the membrane in response to ligand binding to the extracellular domain of FGFR3, we observe no measurable effects of the G380R mutation on FGFR3 ligand-bound dimer configurations. Instead, the most notable effect of the achondroplasia mutation is increased propensity for FGFR3 dimerization in the absence of ligand. This work reveals new information about the molecular events that underlie the achondroplasia phenotype, and highlights differences in FGFR3 activation due to different single amino-acid pathogenic mutations. Copyright © 2016 Elsevier B.V. All rights reserved.

Wide-field lifetime-based FRET imaging for the assessment of early functional distribution of transferrin-based delivery in breast tumor-bearing small animals

Science.gov (United States)

Sinsuebphon, Nattawut; Rudkouskaya, Alena; Barroso, Margarida; Intes, Xavier

2016-02-01

Targeted drug delivery is a critical aspect of successful cancer therapy. Assessment of dynamic distribution of the drug provides relative concentration and bioavailability at the target tissue. The most common approach of the assessment is intensity-based imaging, which only provides information about anatomical distribution. Observation of biomolecular interactions can be performed using Förster resonance energy transfer (FRET). Thus, FRET-based imaging can assess functional distribution and provide potential therapeutic outcomes. In this study, we used wide-field lifetime-based FRET imaging for the study of early functional distribution of transferrin delivery in breast cancer tumor models in small animals. Transferrin is a carrier for cancer drug delivery. Its interaction with its receptor is within a few nanometers, which is suitable for FRET. Alexa Fluor® 700 and Alexa Fluor® 750 were conjugated to holo-transferrin which were then administered via tail vein injection to the mice implanted with T47D breast cancer xenografts. Images were continuously acquired for 60 minutes post-injection. The results showed that transferrin was primarily distributed to the liver, the urinary bladder, and the tumor. The cellular uptake of transferrin, which was indicated by the level of FRET, was high in the liver but very low in the urinary bladder. The results also suggested that the fluorescence intensity and FRET signals were independent. The liver showed increasing intensity and increasing FRET during the observation period, while the urinary bladder showed increasing intensity but minimal FRET. Tumors gave varied results corresponding to their FRET progression. These results were relevant to the biomolecular events that occurred in the animals.

Development of device for grid spring fatigue and a cell-based fuel rod fretting wear tests

International Nuclear Information System (INIS)

Kim, Hyung Kyu; Yoon, Kyung Ho; Kang, Heung Seok; Song, Kee Nam

2001-05-01

As an activity of experimental research on the cause and the remedy of LWR fuel fretting failure, developed is test equipment for fatigue of grid spring and cell-based fuel rod fretting wear test. The equipment enables to perform the fretting wear test in the case of gap existence between spring and cladding, which has not been possible by the previously developed one (KAERI/TR-1570/2000). It can also provide fatigue test capability with the frequency of more than 10 Hz. Used are a servo-motor, an eccentric cylinder and lever mechanism for driving system as was similarly used for the previous equipment. In fretting wear test, up to 2 span-length of a fuel cladding tube can be accommodated. For fatigue test, on the other hand, a device for clamping the spring fixture is installed additionally. As a feature of the present equipment, the gap or the contacting force between a spring and a tube can be adjusted during the fretting wear test, while an initial spring force can be simulated for the fatigue test. Tests will be conducted in air at room temperature. In this report, every part of the equipment is explained with photographs, which will provide an easy understanding. Test procedure such as specimen installation, sequence of operation and program handling is also given. As a performance test of the present equipment, displacement range is measured when the hinge of the lever locates at its maximum and minimum positions. This will be used as basic information when additional eccentric cylinder is necessary for different displacement ranges

Ratiometric Fluorescent Detection of Pb2+ by FRET-Based Phthalocyanine-Porphyrin Dyads.

Science.gov (United States)

Zhang, Dongli; Zhu, Mengliang; Zhao, Luyang; Zhang, Jinghui; Wang, Kang; Qi, Dongdong; Zhou, Yang; Bian, Yongzhong; Jiang, Jianzhuang

2017-12-04

Sensitive and selective detection of Pb 2+ is a very worthwhile endeavor in terms of both human health and environmental protection, as the heavy metal is fairly ubiquitous and highly toxic. In this study, we designed phthalocyanine-porphyrin (Pc-Por) heterodyads, namely, H 2 Pc-α-ZnPor (1) and H 2 Pc-β-ZnPor (2), by connecting a zinc(II) porphyrin moiety to the nonperipheral (α) or peripheral (β) position of a metal-free phthalocyanine moiety. Upon excitation at the porphyrin Soret region (420 nm), both of the dyads exhibited not only a porphyrin emission (605 nm) but also a phthalocyanine emission (ca. 700 nm), indicating the occurrence of intramolecular fluorescence resonance energy transfer (FRET) processes from the porphyrin donor to the phthalocyanine acceptor. The dyads can selectively bind Pb 2+ in the phthalocyanine core leading to a red shift of the phthalocyanine absorption and thus a decrease of spectral overlap between the porphyrin emission and phthalocyanine absorption, which in turn suppresses the intramolecular FRET. In addition, the binding of Pb 2+ can highly quench the emission of phthalocyanine by heavy-metal ion effects. The synergistic coupled functions endow the dyads with remarkable ratiometric fluorescent responses at two distinct wavelengths (F 605 /F 703 for 1 and F 605 /F 700 for 2). The emission intensity ratio increased as a linear function to the concentration of Pb 2+ in the range of 0-4.0 μM, whereas the detection limits were determined to be 3.4 × 10 -9 and 2.2 × 10 -8 M for 1 and 2, respectively. Furthermore, by comparative study of 1 and 2, the effects of distance and relative orientation between Pc and ZnPor fluorophores on the FRET efficiency and sensing performance were highlighted, which is helpful for further optimizing such FRET systems.

Characterization of membrane protein interactions in plasma membrane derived vesicles with quantitative imaging Förster resonance energy transfer.

Science.gov (United States)

Sarabipour, Sarvenaz; Del Piccolo, Nuala; Hristova, Kalina

2015-08-18

Here we describe an experimental tool, termed quantitative imaging Förster resonance energy transfer (QI-FRET), that enables the quantitative characterization of membrane protein interactions. The QI-FRET methodology allows us to acquire binding curves and calculate association constants for complex membrane proteins in the native plasma membrane environment. The method utilizes FRET detection, and thus requires that the proteins of interest are labeled with florescent proteins, either FRET donors or FRET acceptors. Since plasma membranes of cells have complex topologies precluding the acquisition of two-dimensional binding curves, the FRET measurements are performed in plasma membrane derived vesicles that bud off cells as a result of chemical or osmotic stress. The results overviewed here are acquired in vesicles produced with an osmotic vesiculation buffer developed in our laboratory, which does not utilize harsh chemicals. The concentrations of the donor-labeled and the acceptor-labeled proteins are determined, along with the FRET efficiencies, in each vesicle. The experiments utilize transient transfection, such that a wide variety of concentrations is sampled. Then, data from hundreds of vesicles are combined to yield dimerization curves. Here we discuss recent findings about the dimerization of receptor tyrosine kinases (RTKs), membrane proteins that control cell growth and differentiation via lateral dimerization in the plasma membrane. We focus on the dimerization of fibroblast growth factor receptor 3 (FGFR3), a RTK that plays a critically important role in skeletal development. We study the role of different FGFR3 domains in FGFR3 dimerization in the absence of ligand, and we show that FGFR3 extracellular domains inhibit unliganded dimerization, while contacts between the juxtamembrane domains, which connect the transmembrane domains to the kinase domains, stabilize the unliganded FGFR3 dimers. Since FGFR3 has been documented to harbor many pathogenic

Fretting corrosion tests on orthopedic plates and screws made of ASTM F138 stainless steel

OpenAIRE

Santos,Claudio Teodoro dos; Barbosa,Cássio; Monteiro,Maurício de Jesus; Abud,Ibrahim de Cerqueira; Caminha,Ieda Maria Vieira; Roesler,Carlos Rodrigo de Mello

2015-01-01

Introduction Although there has been significant progress in the design of implants for osteosynthesis, the occurrence of failures in these medical devices are still frequent. These implants are prone to suffer from fretting corrosion due to micromotion that takes place between the screw heads and plate holes. Consequently, fretting corrosion has been the subject of research in order to understand its influence on the structural integrity of osteosynthesis implants. The aim of this paper is t...

Fretting Wear Behaviors of Aluminum Cable Steel Reinforced (ACSR Conductors in High-Voltage Transmission Line

Directory of Open Access Journals (Sweden)

Xingchi Ma

2017-09-01

Full Text Available This work reports the fretting wear behavior of aluminum cable steel reinforced (ACSR conductors for use in high-voltage transmission line. Fretting wear tests of Al wires were conducted on a servo-controlled fatigue testing machine with self-made assistant apparatus, and their fretting process characteristics, friction force, wear damage, and wear surface morphology were detailed analyzed. The results show that the running regime of Al wires changes from a gross slip regime to a mixed regime more quickly as increasing contact load. With increasing amplitudes, gross slip regimes are more dominant under contact loads of lower than 30 N. The maximum friction force is relatively smaller in the NaCl solution than in a dry friction environment. The primary wear mechanisms in dry friction environments are abrasive wear and adhesive wear whereas abrasive wear and fatigue damage are dominant in NaCl solution.

Detection of protease activity by fluorescent protein FRET sensors: from computer simulation to live cells

Science.gov (United States)

Goryashchenko, Alexander S.; Khrenova, Maria G.; Savitsky, Alexander P.

2018-04-01

Förster resonance energy transfer (FRET) sensors are widely used for the detection of protease activity in vitro and in vivo. Usually they consist of a FRET pair connected with a polypeptide linker containing a specific cleavage site for the relevant protease. Use of the fluorescent proteins as components of the FRET pair allows genetic encoding of such sensors and solves the problem of their delivery into live cells and animals. There are several ways to improve the properties of such sensors, mainly to increase FRET efficiency and therefore the dynamic range. One of the ways to achieve this is to use a non-fluorescent chromoprotein as an acceptor. Molecular dynamic simulations may assist in the construction of linker structures connecting donor and acceptor molecules. Estimation of the orientation factor κ 2 can be obtained by methods based on quantum theory and combined quantum mechanics/molecular mechanics approaches. The linker can be structured by hydrophobic interactions, bringing it into a closed conformation that shortens the distance between donor and acceptor and, consequently, increases FRET efficiency. We analyzed the effects of different linker structures on the detection of caspase-3 activity using a non-fluorescent acceptor. Also we have constructed the Tb3+- TagRFP sensor in which a complex of the terbium ion and terbium-binding peptide is used as a donor. This allowed us to use the unique property of lanthanide ions—fluorescence lifetime up to milliseconds—to perform measurements with time delay and exclude the nanosecond-order fluorescence. Using our systems as a starting point, by changing the recognition site in the linker it is possible to perform imaging of different protease activity in vitro or in vivo.

Ratiometric FRET-based detection of DNA and micro-RNA in solution

International Nuclear Information System (INIS)

Matveeva, Evgenia G.; Gryczynski, Zygmunt; Stewart, Donald R.; Gryczynski, Ignacy

2009-01-01

A ratiometric method for detecting DNA oligomers in bulk solution based on Foerster resonance energy transfer (FRET) is described. The two fluorescence signals (green and red), originating from Cy3 (donor, green) and Cy5 (acceptor, red) labels, are simultaneously detected from the pre-hybridized Cy3oligomerY:Cy5oligomerX system. The ratio of red to green intensities is sensitive to the presence of the single-stranded complimentary oligomer, which replaces single-stranded Cy3oligomerY in the donor:acceptor complex and perturbs the FRET. The detection scheme is generally applicable to the detection of DNA and RNA, and particularly micro-RNA. The proposed method is applicable to various double-stranded various lengths targets (manipulation of the sample preparation conditions, such as temperature, incubation time, denaturizing agent, may be needed).

Flow-induced vibration and fretting-wear damage in a moisture separator reheater

International Nuclear Information System (INIS)

Pettigrew, M.J.; Taylor, C.E.; Fisher, N.J.

1996-01-01

Tube failures due to excessive flow-induced vibration were experienced in the tube bundles of moisture separator reheaters in a BWR nuclear station. This paper presents the results of a root cause analysis and covers recommendations for continued operation and for replacement tube bundles. The following tasks are discussed: tube failure analysis; flow velocity distribution calculations; flow-induced vibration analysis with particular emphasis on finned-tubes; fretting-wear testing of a tube and tube-support material combination under simulated operating conditions; field measurements of flow-induced vibration; and development of vibration specifications for replacement tube bundles. The effect of transient operating conditions and of other operational changes such as tube fouling were considered in the analysis. This paper outlines a typical field problem and illustrates the application of flow-induced vibration technology for the solution of a practical problem

Electroporation and microinjection successfully deliver single-stranded and duplex DNA into live cells as detected by FRET measurements.

Directory of Open Access Journals (Sweden)

Rosemary A Bamford

Full Text Available Förster resonance energy transfer (FRET technology relies on the close proximity of two compatible fluorophores for energy transfer. Tagged (Cy3 and Cy5 complementary DNA strands forming a stable duplex and a doubly-tagged single strand were shown to demonstrate FRET outside of a cellular environment. FRET was also observed after transfecting these DNA strands into fixed and live cells using methods such as microinjection and electroporation, but not when using lipid based transfection reagents, unless in the presence of the endosomal acidification inhibitor bafilomycin. Avoiding the endocytosis pathway is essential for efficient delivery of intact DNA probes into cells.

FRET microscopy autologous tumor lysate processing in mature dendritic cell vaccine therapy

Directory of Open Access Journals (Sweden)

Ridolfi Ruggero

2010-06-01

Full Text Available Abstract Background Antigen processing by dendritic cells (DC exposed to specific stimuli has been well characterized in biological studies. Nonetheless, the question of whether autologous whole tumor lysates (as used in clinical trials are similarly processed by these cells has not yet been resolved. Methods In this study, we examined the transfer of peptides from whole tumor lysates to major histocompatibility complex class II molecules (MHC II in mature dendritic cells (mDC from a patient with advanced melanoma. Tumor antigenic peptides-MHC II proximity was revealed by Förster Resonance Energy Transfer (FRET measurements, which effectively extends the application of fluorescence microscopy to the molecular level ( Results We detected significant energy transfer between donor and acceptor-labelled antibodies against HLA-DR at the membrane surface of mDC. FRET data indicated that fluorescent peptide-loaded MHC II molecules start to accumulate on mDC membranes at 16 hr from the maturation stimulus, steeply increasing at 22 hr with sustained higher FRET detected up to 46 hr. Conclusions The results obtained imply that the patient mDC correctly processed the tumor specific antigens and their display on the mDC surface may be effective for several days. These observations support the rationale for immunogenic efficacy of autologous tumor lysates.

Fretting Wear Damage Mechanism of Uranium under Various Atmosphere and Vacuum Conditions

Directory of Open Access Journals (Sweden)

Zhengyang Li

2018-04-01

Full Text Available A fretting wear experiment with uranium has been performed on a linear reciprocating tribometer with ball-on-disk contact. This study focused on the fretting behavior of the uranium under different atmospheres (Ar, Air (21% O2 + 78% N2, and O2 and vacuum conditions (1.05 and 1 × 10−4 Pa. Evolution of friction was assessed by coefficient of friction (COF and friction-dissipated energy. The oxide of the wear surface was evaluated by Raman spectroscopy. The result shows that fretting wear behavior presents strong atmosphere and vacuum condition dependence. With increasing oxygen content, the COF decreases due to abrasive wear and formation of oxide film. The COF in the oxygen condition is at least 0.335, and it has a maximum wear volume of about 1.48 × 107 μm3. However, the COF in a high vacuum condition is maximum about 1.104, and the wear volume is 1.64 × 106 μm3. The COF in the low vacuum condition is very different: it firstly increased and then decreased rapidly to a steady value. It is caused by slight abrasive wear and the formation of tribofilm after thousands of cycles.

Fretting fatigue life estimation using fatigue damage gradient correction factor in various contact configurations

Energy Technology Data Exchange (ETDEWEB)

Hwang, Dong Hyeon; Cho, Sung-San [Hongik University, Seoul (Korea, Republic of)

2017-03-15

A fretting fatigue life estimation method that takes into account the stress gradient effect was developed by the authors [Journal of Mechanical Science and Technology, 28 (2014) 2153-2159]. In the developed method, fatigue damage value at the cracking location is corrected with fatigue damage gradient and the corrected value is compared directly with the plain fatigue data for life estimation. In other words, the correction factor is the ratio of plain fatigue damage to fretting fatigue damage at the same life and a function of fatigue damage gradient. Since reliability of the method was verified only for cylinder-on-flat contact configuration in the previous study, the present study extends application of the method to flat-on-flat contact configurations by developing the correction factor for both the contact configuration. Fretting fatigue experiments were conducted to obtain fatigue life data for various fretting pads. Finite element analyses were conducted to evaluate the Smith-Watson-Topper (SWT) fatigue damage parameter in the cracking region. It is revealed that the SWT parameter in fat-on-flat contact configuration decreases exponentially away from the surface as in cylinder-on-flat contact configuration, and thus the SWT gradient at the surface can be evaluated reliably. Moreover, it is found that decrease in the SWT parameter around the cracking location can be expressed by piecewise exponential curves. If the gradient of SWT at the surface is used as a representative value of SWT gradient, it is impossible to establish functional relationship between the SWT gradient and the correction factor for both the contact configurations although it was possible for cylinder-on-flat contact configuration. However, if weighted average of the SWT gradient values obtained from each exponential curve in the piecewise exponential curve is used as a representative value, the correction factor for both the contact configurations becomes a function of the SWT gradient

Development of a Fluorescence Resonance Energy Transfer (FRET)-Based DNA Biosensor for Detection of Synthetic Oligonucleotide of Ganoderma boninense.

Science.gov (United States)

Mohd Bakhori, Noremylia; Yusof, Nor Azah; Abdullah, Abdul Halim; Hussein, Mohd Zobir

2013-12-01

An optical DNA biosensor based on fluorescence resonance energy transfer (FRET) utilizing synthesized quantum dot (QD) has been developed for the detection of specific-sequence of DNA for Ganoderma boninense, an oil palm pathogen. Modified QD that contained carboxylic groups was conjugated with a single-stranded DNA probe (ssDNA) via amide-linkage. Hybridization of the target DNA with conjugated QD-ssDNA and reporter probe labeled with Cy5 allows for the detection of related synthetic DNA sequence of Ganoderma boninense gene based on FRET signals. Detection of FRET emission before and after hybridization was confirmed through the capability of the system to produce FRET at 680 nm for hybridized sandwich with complementary target DNA. No FRET emission was observed for non-complementary system. Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system. The developed biosensor has shown high sensitivity with detection limit of 3.55 × 10(-9) M. TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA. This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

Development of a Fluorescence Resonance Energy Transfer (FRET-Based DNA Biosensor for Detection of Synthetic Oligonucleotide of Ganoderma boninense

Directory of Open Access Journals (Sweden)

Noremylia Mohd Bakhori

2013-12-01

Full Text Available An optical DNA biosensor based on fluorescence resonance energy transfer (FRET utilizing synthesized quantum dot (QD has been developed for the detection of specific-sequence of DNA for Ganoderma boninense, an oil palm pathogen. Modified QD that contained carboxylic groups was conjugated with a single-stranded DNA probe (ssDNA via amide-linkage. Hybridization of the target DNA with conjugated QD-ssDNA and reporter probe labeled with Cy5 allows for the detection of related synthetic DNA sequence of Ganoderma boninense gene based on FRET signals. Detection of FRET emission before and after hybridization was confirmed through the capability of the system to produce FRET at 680 nm for hybridized sandwich with complementary target DNA. No FRET emission was observed for non-complementary system. Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system. The developed biosensor has shown high sensitivity with detection limit of 3.55 × 10−9 M. TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA. This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

A dansyl-rhodamine ratiometric fluorescent probe for Hg2+ based on FRET mechanism.

Science.gov (United States)

Xie, Puhui; Guo, Fengqi; Wang, Lingyu; Yang, Sen; Yao, Denghui; Yang, Guoyu

2015-03-01

Based on resonance energy transfer (FRET) from dansyl to rhodamine 101, a new fluorescent probe (compound 1) containing rhodamine 101 and a dansyl unit was synthesized for detecting Hg(2+) through ratiometric sensing in DMSO aqueous solutions. This probe shows a fast, reversible and selective response toward Hg(2+) in a wide pH range. Hg(2+) induced ring-opening reactions of the spirolactam rhodamine moiety of 1, leading to the formation of fluorescent derivatives that can serve as the FRET acceptors. Very large stokes shift (220 nm) was observed in this case. About 97-fold increase in fluorescence intensity ratio was observed upon its binding with Hg(2+).

Experimental studies of resistance fretting-wear of fuel rods for VVER-1000 and TVS-KVADRAT fuel assemblies

International Nuclear Information System (INIS)

Makarov, V.; Afanasiev, A.; Egorov, Yu.; Matvienko, I.

2015-01-01

The paper covers the results of the studies performed to justify the wear resistance of fuel rods in contact with the spacer grids of TVS VVER-1000 fuel assembly and TVS-KVADRAT square fuel assembly of Russian design for PWR-900 reactor. The presented results of three testing stages comprise: Testing of mockup fuel rods of VVER TVS fuel assembly for fretting wear under the conditions of the water chemistry of VVER reactor; Testing models of different design embodiments of the fuel rods for VVER TVS fuel assembly for fretting wear in still cold water; Testing mockup fuel rods of TVS-KVADRAT square fuel assembly for PWR reactor for frettingwear under the conditions of PWR water chemistry. The effect of structural and operational factors was determined (amplitudes, fuel rod vibration frequencies, values of cladding-to-spacer grid cell gap for the depth of fuel rod cladding wear etc.), an assessment was made of the threshold values of fuel rod vibration parameters, which, if not exceeded, provide the absence of the fuel rod cladding fretting wear in the fuel rod-to spacer grid contact area. Key words: fretting wear, fuel rod, spacer grid, VVER, PWR (author)

[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.

NIR FRET Fluorophores for Use as an Implantable Glucose Biosensor

Directory of Open Access Journals (Sweden)

Majed DWEIK

2008-12-01

Full Text Available Development of an in vivo optical sensor requires the utilization of Near Infra Red (NIR fluorophores due to their ability to operate within the biological tissue window. Alexa Fluor 750 (AF750 and Alexa Fluor 680 (AF680 were examined as potential NIR fluorophores for an in vivo fluorescence resonance energy transfer (FRET glucose biosensor. AF680 and AF750 found to be a FRET pair and percent energy transfer was calculated. Next, the tested dye pair was utilized in a competitive binding assay in order to detect glucose. Concanavalin A (Con A and dextran have binding affinity, but in the presence of glucose, glucose displaces dextran due to its higher affinity to Con A than dextran. Finally, the percent signal transfer through porcine skin was examined. The results showed with approximately 4.0 mm porcine skin thickness, 1.98 % of the fluorescence was transmitted and captured by the detector.

Fretting wear of steam generator tubes: high-temperature tests on AECL rig

International Nuclear Information System (INIS)

Guerout, F.; Zbinden, M.

1993-07-01

The R and DD has undertaken the study of fretting-wear of Alloy 600 S.G. tubes which occurs by contact with migrating items. The test series was performed in Canada at AECL Research (Atomic Energy of Canada Limited) as part of an exchange program. Four types of configuration were envisaged: a tube-to-drilled hole support contact which provides reference results and three types of tube-to-support contacts which simulate the tube fretting-wear induced by a welding rod, a threaded rod and a knife-edge rod support. This programme is completed by the study of the contact between a S.G. tube and a neighbouring S.G. tube which has been broken after plugging. (authors). 1 tab., 3 refs

FRET study of membrane proteins: determination of the tilt and orientation of the N-terminal domain of M13 major coat protein

NARCIS (Netherlands)

Nazarov, P.V.; Koehorst, R.B.M.; Vos, W.L.; Apanasovich, V.V.; Hemminga, M.A.

2007-01-01

A formalism for membrane protein structure determination was developed. This method is based on steady-state FRET data and information about the position of the fluorescence maxima on site-directed fluorescent labeled proteins in combination with global data analysis utilizing simulation-based

Accuracy of maximum likelihood estimates of a two-state model in single-molecule FRET

Energy Technology Data Exchange (ETDEWEB)

Gopich, Irina V. [Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland 20892 (United States)

2015-01-21

Photon sequences from single-molecule Förster resonance energy transfer (FRET) experiments can be analyzed using a maximum likelihood method. Parameters of the underlying kinetic model (FRET efficiencies of the states and transition rates between conformational states) are obtained by maximizing the appropriate likelihood function. In addition, the errors (uncertainties) of the extracted parameters can be obtained from the curvature of the likelihood function at the maximum. We study the standard deviations of the parameters of a two-state model obtained from photon sequences with recorded colors and arrival times. The standard deviations can be obtained analytically in a special case when the FRET efficiencies of the states are 0 and 1 and in the limiting cases of fast and slow conformational dynamics. These results are compared with the results of numerical simulations. The accuracy and, therefore, the ability to predict model parameters depend on how fast the transition rates are compared to the photon count rate. In the limit of slow transitions, the key parameters that determine the accuracy are the number of transitions between the states and the number of independent photon sequences. In the fast transition limit, the accuracy is determined by the small fraction of photons that are correlated with their neighbors. The relative standard deviation of the relaxation rate has a “chevron” shape as a function of the transition rate in the log-log scale. The location of the minimum of this function dramatically depends on how well the FRET efficiencies of the states are separated.

Fretting-wear characteristics of steam generator tubes contacting with foreign object

International Nuclear Information System (INIS)

Jo, Jong Chull; Jhung, Myung Jo; Kim, Woong Sik; Kim, Hho Jung

2003-01-01

Fretting-wear characteristics of steam generator tubes contacting with foreign object has been investigated in this study. The operating steam generator shell-side flow field conditions are obtained from three-dimensional steam generator flow calculation using a well-validated steam generator thermal-hydraulic analysis computer code. Modal analyses are performed for the finite element modelings of tubes to get the natural frequency, corresponding mode shape and participation factor. The wear rate of a steam generator tube caused by foreign object is calculated using the Archard formula and the remaining life of the tube is predicted. In addition, the effects of internal pressure and flow velocity on the remaining life of the tube are discussed in this paper

Intonation and compensation of fretted string instruments

Science.gov (United States)

Varieschi, Gabriele; Gower, Christina

2011-04-01

We discuss theoretical and physical models that are useful for analyzing the intonation of musical instruments such as guitars and mandolins and can be used to improve the tuning on these instruments. The placement of frets on the fingerboard is designed according to mathematical rules and the assumption of an ideal string. The analysis becomes more complicated when we include the effects of deformation of the string and inharmonicity due to other string characteristics. As a consequence, perfect intonation of all the notes on the instrument cannot be achieved, but complex compensation procedures can be introduced to minimize the problem. To test the validity of these procedures, we performed extensive measurements using standard monochord sonometers and other acoustical devices, confirming the correctness of our theoretical models. These experimental activities can be integrated into acoustics courses and laboratories and can become a more advanced version of basic experiments with monochords and sonometers. This work was supported by a grant from the Frank R. Seaver College of Science and Engineering, Loyola Marymount University.

Preliminary Study on the Fretting Wear Behaviors of a Duel Cooled Fuel Rod

Energy Technology Data Exchange (ETDEWEB)

Lee, Y.H.; Lee, K.H.; Kim, H.K. [KAERI, 150 Dukjin-dong Yuseon-gu Daejeon, 305-353 (Korea, Republic of)

2009-06-15

Based on MIT's concept, an innovative fuel development project was launched by KAERI that a substantial power up-rating could be realized by introducing an internally and externally double cooled annular fuel for current PWR reactors. In order to apply this duel cooled fuel to an OPR 1000 reactor system, geometrical features of structural parts in a fuel assembly should be changed except an overall dimension of a fuel assembly. Typical changes are summarized as fuel rod diameter and weight, shape and position of a spacer grid spring, etc. When considering a duel cooled fuel rod, its vibration characteristic and fretting behavior should be verified because the modified shape and dimension of spacer grid spring, fuel rod diameter and weight, number of spacer grid assembly are closely related to a flow-induced vibration in a duel cooled fuel assembly. In this study, based on FIV test results of 4x4 fuel assembly, fretting wear tests of an outer duel cooled fuel rod were performed by using an embossing type spacer grid spring that could adjust its spring stiffness. The discussion was focused on the evaluation of the optimized spring stiffness and spring position in 1x1 cell by analyzing the fretting wear results. (authors)

Quantitative analysis of receptor imaging

International Nuclear Information System (INIS)

Fu Zhanli; Wang Rongfu

2004-01-01

Model-based methods for quantitative analysis of receptor imaging, including kinetic, graphical and equilibrium methods, are introduced in detail. Some technical problem facing quantitative analysis of receptor imaging, such as the correction for in vivo metabolism of the tracer and the radioactivity contribution from blood volume within ROI, and the estimation of the nondisplaceable ligand concentration, is also reviewed briefly

Diagnostic performance of semi-quantitative and quantitative stress CMR perfusion analysis: a meta-analysis.

Science.gov (United States)

van Dijk, R; van Assen, M; Vliegenthart, R; de Bock, G H; van der Harst, P; Oudkerk, M

2017-11-27

Stress cardiovascular magnetic resonance (CMR) perfusion imaging is a promising modality for the evaluation of coronary artery disease (CAD) due to high spatial resolution and absence of radiation. Semi-quantitative and quantitative analysis of CMR perfusion are based on signal-intensity curves produced during the first-pass of gadolinium contrast. Multiple semi-quantitative and quantitative parameters have been introduced. Diagnostic performance of these parameters varies extensively among studies and standardized protocols are lacking. This study aims to determine the diagnostic accuracy of semi- quantitative and quantitative CMR perfusion parameters, compared to multiple reference standards. Pubmed, WebOfScience, and Embase were systematically searched using predefined criteria (3272 articles). A check for duplicates was performed (1967 articles). Eligibility and relevance of the articles was determined by two reviewers using pre-defined criteria. The primary data extraction was performed independently by two researchers with the use of a predefined template. Differences in extracted data were resolved by discussion between the two researchers. The quality of the included studies was assessed using the 'Quality Assessment of Diagnostic Accuracy Studies Tool' (QUADAS-2). True positives, false positives, true negatives, and false negatives were subtracted/calculated from the articles. The principal summary measures used to assess diagnostic accuracy were sensitivity, specificity, andarea under the receiver operating curve (AUC). Data was pooled according to analysis territory, reference standard and perfusion parameter. Twenty-two articles were eligible based on the predefined study eligibility criteria. The pooled diagnostic accuracy for segment-, territory- and patient-based analyses showed good diagnostic performance with sensitivity of 0.88, 0.82, and 0.83, specificity of 0.72, 0.83, and 0.76 and AUC of 0.90, 0.84, and 0.87, respectively. In per territory

One-step synthesis of DNA functionalized cadmium-free quantum dots and its application in FRET-based protein sensing

Energy Technology Data Exchange (ETDEWEB)

Zhang, Cuiling, E-mail: clzhang@chem.ecnu.edu.cn [Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241 (China); Ding, Caiping [Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241 (China); Zhou, Guohua [School of Chemistry and Chemical Engineering, Lingnan Normal University, Zhanjiang, 524048 (China); Xue, Qin [Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241 (China); Xian, Yuezhong, E-mail: yzxian@chem.ecnu.edu.cn [Department of Chemistry, School of Chemistry and Molecular Engineering, East China Normal University, Shanghai 200241 (China)

2017-03-08

DNA functionalized quantum dots (QDs) are promising nanoprobes for the fluorescence resonance energy transfer (FRET)-based biosensing. Herein, cadmium-free DNA functionalized Mn-doped ZnS (DNA-ZnS:Mn{sup 2+}) QDs were successfully synthesized by one-step route. As-synthesized QDs show excellent photo-stability with the help of PAA and DNA. Then, we constructed a novel FRET model based on the QDs and WS{sub 2} nanosheets as the energy donor-acceptor pairs, which was successfully applied for the protein detection through the terminal protection of small molecule-linked DNA assay. This work not only explores the potential bioapplication of the DNA-ZnS:Mn{sup 2+} QDs, but also provides a platform for the investigation of small molecule-protein interaction. - Highlights: • The stable and cadmium-free DNA functionalized ZnS:Mn{sup 2+} QDs were successfully synthesized through a facile one-step route. • We constructed a novel FRET system based on one-step synthesized DNA-ZnS:Mn{sup 2+} QDs (donor) and WS{sub 2} nanosheets (acceptor). • The FRET-based strategy was applied for the detection of streptavidin and folate receptor by combining TPSMLD and Exo III.

Decoupling of size and shape fluctuations in heteropolymeric sequences reconciles discrepancies in SAXS vs. FRET measurements.

Science.gov (United States)

Fuertes, Gustavo; Banterle, Niccolò; Ruff, Kiersten M; Chowdhury, Aritra; Mercadante, Davide; Koehler, Christine; Kachala, Michael; Estrada Girona, Gemma; Milles, Sigrid; Mishra, Ankur; Onck, Patrick R; Gräter, Frauke; Esteban-Martín, Santiago; Pappu, Rohit V; Svergun, Dmitri I; Lemke, Edward A

2017-08-01

Unfolded states of proteins and native states of intrinsically disordered proteins (IDPs) populate heterogeneous conformational ensembles in solution. The average sizes of these heterogeneous systems, quantified by the radius of gyration ( R G ), can be measured by small-angle X-ray scattering (SAXS). Another parameter, the mean dye-to-dye distance ( R E ) for proteins with fluorescently labeled termini, can be estimated using single-molecule Förster resonance energy transfer (smFRET). A number of studies have reported inconsistencies in inferences drawn from the two sets of measurements for the dimensions of unfolded proteins and IDPs in the absence of chemical denaturants. These differences are typically attributed to the influence of fluorescent labels used in smFRET and to the impact of high concentrations and averaging features of SAXS. By measuring the dimensions of a collection of labeled and unlabeled polypeptides using smFRET and SAXS, we directly assessed the contributions of dyes to the experimental values R G and R E For chemically denatured proteins we obtain mutual consistency in our inferences based on R G and R E , whereas for IDPs under native conditions, we find substantial deviations. Using computations, we show that discrepant inferences are neither due to methodological shortcomings of specific measurements nor due to artifacts of dyes. Instead, our analysis suggests that chemical heterogeneity in heteropolymeric systems leads to a decoupling between R E and R G that is amplified in the absence of denaturants. Therefore, joint assessments of R G and R E combined with measurements of polymer shapes should provide a consistent and complete picture of the underlying ensembles.

The synergy of corrosion and fretting wear process on Inconel 690 in the high temperature high pressure water environment

Science.gov (United States)

Wang, Zihao; Xu, Jian; Li, Jie; Xin, Long; Lu, Yonghao; Shoji, Tetsuo; Takeda, Yoichi; Otsuka, Yuichi; Mutoh, Yoshiharu

2018-04-01

The synergistic effect of corrosion and fretting process of the steam generator (SG) tube was investigated by using a self-designed high temperature test rig in this paper. The experiments were performed at 100°C , 200°C and 288°C , respectively. The fretting corrosion damage was studied by optical microscopy (OM), scanning electron microscope (SEM), energy dispersive spectrometer (EDS), Raman spectroscopy and auger electron spectroscopy (AES). The results demonstrated that the corrosion process in high temperature high pressure (HTHP) water environment had a distinct interaction with the fretting process of Inconel 690. With the increment of temperature, the damage mechanism changed from a simple mechanical process to a mechanochemical process.

Friction and wear properties of high-velocity oxygen fuel sprayed WC-17Co coating under rotational fretting conditions

Science.gov (United States)

Luo, Jun; Cai, Zhenbing; Mo, Jiliang; Peng, Jinfang; Zhu, Minhao

2016-05-01

Rotational fretting which exist in many engineering applications has incurred enormous economic loss. Thus, accessible methods are urgently needed to alleviate or eliminate damage by rotational fretting. Surface engineering is an effective approach that is successfully adopted to enhance the ability of components to resist the fretting damage. In this paper, using a high-velocity oxygen fuel sprayed (HVOF) technique WC-17Co coating is deposited on an LZ50 steel surface to study its properties through Vickers hardness testing, scanning electric microscope (SEM), energy dispersive X-ray spectroscopy (EDX), and X-ray diffractrometry (XRD). Rotational fretting wear tests are conducted under normal load varied from 10 N to 50 N, and angular displacement amplitudes vary from 0.125° to 1°. Wear scars are examined using SEM, EDX, optical microscopy (OM), and surface topography. The experimental results reveal that the WC-17Co coating adjusted the boundary between the partial slip regime (PSR) and the slip regime (SR) to the direction of smaller amplitude displacement. As a result, the coefficients of friction are consistently lower than the substrate's coefficients of friction both in the PSR and SR. The damage to the coating in the PSR is very slight. In the SR, the coating exhibits higher debris removal efficiency and load-carrying capacity. The bulge is not found for the coating due to the coating's higher hardness to restrain plastic flow. This research could provide experimental bases for promoting industrial application of WC-17Co coating in prevention of rotational fretting wear.

The Effect of Modulation Ratio of Cu/Ni Multilayer Films on the Fretting Damage Behaviour of Ti-811 Titanium Alloy.

Science.gov (United States)

Zhang, Xiaohua; Liu, Daoxin; Li, Xiaoying; Dong, Hanshan; Xi, Yuntao

2017-05-26

To improve the fretting damage (fretting wear and fretting fatigue) resistance of Ti-811 titanium alloy, three Cu/Ni multilayer films with the same modulation period thickness (200 nm) and different modulation ratios (3:1, 1:1, 1:3) were deposited on the surface of the alloy via ion-assisted magnetron sputtering deposition (IAD). The bonding strength, micro-hardness, and toughness of the films were evaluated, and the effect of the modulation ratio on the room-temperature fretting wear (FW) and fretting fatigue (FF) resistance of the alloy was determined. The results indicated that the IAD technique can be successfully used to prepare Cu/Ni multilayer films, with high bonding strength, low-friction, and good toughness, which yield improved room-temperature FF and FW resistance of the alloy. For the same modulation period (200 nm), the micro-hardness, friction, and FW resistance of the coated alloy increased, decreased, and improved, respectively, with increasing modulation ratio of the Ni-to-Cu layer thickness. However, the FF resistance of the coated alloy increased non-monotonically with the increasing modulation ratio. Among the three Cu/Ni multilayer films, those with a modulation ratio of 1:1 can confer the highest FF resistance to the Ti-811 alloy, owing mainly to their unique combination of good toughness, high strength, and low-friction.

Fretting wear of Inconel 625 at high temperature and in high vacuum

International Nuclear Information System (INIS)

Iwabuchi, A.

1985-01-01

The purpose of this work was to investigate the fretting properties of Inconel 625 at high temperature and in high vacuum. Experiments were carried out under constant conditions with a normal load of 14 N and a peak-to-peak slip amplitude of 110 μm and through 6x10 4 cycles. Several environmental conditions were used. Pressure was varied between 10 -3 and 10 5 Pa at temperatures of 20 and 500 0 C. Temperatures up to 500 0 C were also used at pressures of 10 -3 and 10 5 Pa. At 10 -3 Pa and 500 0 C wear loss was negligible but wear scars showed severe damage consisting of deep cracks and accretion of transferred debris. The coefficient of friction then maintained a high value of 1.7 throughout the fretting test. The critical pressure below which oxidation rate becomes reduced is 10 Pa, a value independent of temperature. At pressures below this critical value the coefficient of friction increases steeply and the fretting mechanism changes from one of oxidative wear to one of adhesive wear. A compacted so-called 'glaze' oxide was formed at temperatures above 300 0 C in air (10 5 Pa) and at pressures above 10 3 Pa at 500 0 C. A comparison of results for Inconel 625 with those for S45C and SUS304 steels and Inconel 600 is given. (orig.)

Homo-FRET Imaging as a tool to quantify protein and lipid clustering

NARCIS (Netherlands)

Bader, A.N.; Hoetzl, S.; Hofman, E.G.; Voortman, J.; van Bergen en Henegouwen, P.M.P.; van Meer, G.; Gerritsen, H.C.

2010-01-01

Homo-FRET, Förster resonance energy transfer between identical fluorophores, can be conveniently measured by observing its effect on the fluorescence anisotropy. This review aims to summarize the possibilities of fluorescence anisotropy imaging techniques to investigate clustering of identical

Noninvasive High-Throughput Single-Cell Analysis of HIV Protease Activity Using Ratiometric Flow Cytometry

Directory of Open Access Journals (Sweden)

Rok Gaber

2013-11-01

Full Text Available To effectively fight against the human immunodeficiency virus infection/ acquired immunodeficiency syndrome (HIV/AIDS epidemic, ongoing development of novel HIV protease inhibitors is required. Inexpensive high-throughput screening assays are needed to quickly scan large sets of chemicals for potential inhibitors. We have developed a Förster resonance energy transfer (FRET-based, HIV protease-sensitive sensor using a combination of a fluorescent protein pair, namely mCerulean and mCitrine. Through extensive in vitro characterization, we show that the FRET-HIV sensor can be used in HIV protease screening assays. Furthermore, we have used the FRET-HIV sensor for intracellular quantitative detection of HIV protease activity in living cells, which more closely resembles an actual viral infection than an in vitro assay. We have developed a high-throughput method that employs a ratiometric flow cytometry for analyzing large populations of cells that express the FRET-HIV sensor. The method enables FRET measurement of single cells with high sensitivity and speed and should be used when subpopulation-specific intracellular activity of HIV protease needs to be estimated. In addition, we have used a confocal microscopy sensitized emission FRET technique to evaluate the usefulness of the FRET-HIV sensor for spatiotemporal detection of intracellular HIV protease activity.

Noninvasive High-Throughput Single-Cell Analysis of HIV Protease Activity Using Ratiometric Flow Cytometry

Science.gov (United States)

Gaber, Rok; Majerle, Andreja; Jerala, Roman; Benčina, Mojca

2013-01-01

To effectively fight against the human immunodeficiency virus infection/acquired immunodeficiency syndrome (HIV/AIDS) epidemic, ongoing development of novel HIV protease inhibitors is required. Inexpensive high-throughput screening assays are needed to quickly scan large sets of chemicals for potential inhibitors. We have developed a Förster resonance energy transfer (FRET)-based, HIV protease-sensitive sensor using a combination of a fluorescent protein pair, namely mCerulean and mCitrine. Through extensive in vitro characterization, we show that the FRET-HIV sensor can be used in HIV protease screening assays. Furthermore, we have used the FRET-HIV sensor for intracellular quantitative detection of HIV protease activity in living cells, which more closely resembles an actual viral infection than an in vitro assay. We have developed a high-throughput method that employs a ratiometric flow cytometry for analyzing large populations of cells that express the FRET-HIV sensor. The method enables FRET measurement of single cells with high sensitivity and speed and should be used when subpopulation-specific intracellular activity of HIV protease needs to be estimated. In addition, we have used a confocal microscopy sensitized emission FRET technique to evaluate the usefulness of the FRET-HIV sensor for spatiotemporal detection of intracellular HIV protease activity. PMID:24287545

A comparison of donor-acceptor pairs for genetically encoded FRET sensors: application to the Epac cAMP sensor as an example.

Directory of Open Access Journals (Sweden)

Gerard N M van der Krogt

Full Text Available We recently reported on CFP-Epac-YFP, an Epac-based single polypeptide FRET reporter to resolve cAMP levels in living cells. In this study, we compared and optimized the fluorescent protein donor/acceptor pairs for use in biosensors such as CFP-Epac-YFP. Our strategy was to prepare a wide range of constructs consisting of different donor and acceptor fluorescent proteins separated by a short linker. Constructs were expressed in HEK293 cells and tested for FRET and other relevant properties. The most promising pairs were subsequently used in an attempt to improve the FRET span of the Epac-based cAMP sensor. The results show significant albeit not perfect correlation between performance in the spacer construct and in the Epac sensor. Finally, this strategy enabled us to identify improved sensors both for detection by sensitized emission and by fluorescent lifetime imaging. The present overview should be helpful in guiding development of future FRET sensors.

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.

Fretting wear behavior of zirconium alloy in B-Li water at 300 °C

Science.gov (United States)

Zhang, Lefu; Lai, Ping; Liu, Qingdong; Zeng, Qifeng; Lu, Junqiang; Guo, Xianglong

2018-02-01

The tangential fretting wear of three kinds of zirconium alloys tube mated with 304 stainless steel (SS) plate was investigated. The tests were conducted in an autoclave containing 300 °C pressurized B-Li water for tube-on-plate contact configuration. The worn surfaces were examined with scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and 3D microscopy. The cross-section of wear scar was examined with transmission electron microscope (TEM). The results indicated that the dominant wear mechanism of zirconium alloys in this test condition was delamination and oxidation. The oxide layer on the fretted area consists of outer oxide layer composed of iron oxide and zirconium oxide and inner oxide layer composed of zirconium oxide.

A Comparative Study on the Formation Mechanism of Wear Scars during the Partial and Full Scale Fretting Wear Tests of Spacer Grids

Energy Technology Data Exchange (ETDEWEB)

Lee, Young Ho; Shin, Chang Hwan; Oh, Dong Seok; Kang, Heung Seok [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2012-05-15

Fretting wear studies for evaluating the contact damages of nuclear fuel rods have been focused on the contact shape, rod motion, contact condition, environment, etc.. However, fretting wear mechanism was dramatically changed with slight variation of test variables such as test environments and contact shapes. For example, in an unlubricated condition, effects of wear debris and/or its layer on the fretting wear mechanism showed that the formation of a well-developed layer on the contact surfaces has a beneficial effect for decreasing a friction coefficient. Otherwise, a severe wear was happened due to a third body abrasion. In addition, in water lubrication condition, some of wear debris was remained on worn surface of fuel rod specimens at both sliding and impacting loading conditions. So, it is apparent that a wear rate of fuel rod specimen was easily accelerated by the third-body abrasion. This is because the restrained agglomeration behavior between generated wear particles results in rapid removal of wear debris and its layer. In case of contact shape effects, previous studies show that wear debris are easily trapped between contact surfaces and its debris layer was well developed in a localized area especially in a concave spring rather than a convex spring shape. Consequently, localized wear was happened at both ends of a concave spring and center region of a convex spring. So, it is useful for determining the fretting wear resistance of spacer gird spring and dimple by using part unit in the various lubricated conditions. It is well known that the fretting wear phenomenon of nuclear fuel rod is originated from a flow-induced vibration (FIV) due to the rapid primary coolant. This means that both rod vibration and debris removal behavior were affected by flow fields around the contact regions between fuel rod and spring/dimple. However, all most of the fretting tests were performed by simulating rod vibrating motions such as axial vibration, conservative rod

A Comparative Study on the Formation Mechanism of Wear Scars during the Partial and Full Scale Fretting Wear Tests of Spacer Grids

International Nuclear Information System (INIS)

Lee, Young Ho; Shin, Chang Hwan; Oh, Dong Seok; Kang, Heung Seok

2012-01-01

Fretting wear studies for evaluating the contact damages of nuclear fuel rods have been focused on the contact shape, rod motion, contact condition, environment, etc.. However, fretting wear mechanism was dramatically changed with slight variation of test variables such as test environments and contact shapes. For example, in an unlubricated condition, effects of wear debris and/or its layer on the fretting wear mechanism showed that the formation of a well-developed layer on the contact surfaces has a beneficial effect for decreasing a friction coefficient. Otherwise, a severe wear was happened due to a third body abrasion. In addition, in water lubrication condition, some of wear debris was remained on worn surface of fuel rod specimens at both sliding and impacting loading conditions. So, it is apparent that a wear rate of fuel rod specimen was easily accelerated by the third-body abrasion. This is because the restrained agglomeration behavior between generated wear particles results in rapid removal of wear debris and its layer. In case of contact shape effects, previous studies show that wear debris are easily trapped between contact surfaces and its debris layer was well developed in a localized area especially in a concave spring rather than a convex spring shape. Consequently, localized wear was happened at both ends of a concave spring and center region of a convex spring. So, it is useful for determining the fretting wear resistance of spacer gird spring and dimple by using part unit in the various lubricated conditions. It is well known that the fretting wear phenomenon of nuclear fuel rod is originated from a flow-induced vibration (FIV) due to the rapid primary coolant. This means that both rod vibration and debris removal behavior were affected by flow fields around the contact regions between fuel rod and spring/dimple. However, all most of the fretting tests were performed by simulating rod vibrating motions such as axial vibration, conservative rod

Stability Loss of the Cemented Stem of Hip Prosthesis due to Fretting Corrosion Fatigue

Directory of Open Access Journals (Sweden)

L. Capitanu

2017-12-01

Full Text Available Aim of this project was to study the fretting behaviour of the cemented femoral stem fixation of a total hip prosthesis, trying to capture the loss of contact between the femoral stem and polymetylmethacrilate cement fixation. To have a landmark, studies were performed compared with cementless fixation, where no fretting phenomenon occurs, on real prostheses, under biological 3D loading conditions. A fatigue test device, installed on a servo-hydraulic triaxial dynamic testing machine was used. It allowed monitoring the flexion-extension, abduction-adduction, inner-outer rotation movements, and the variation of the torsional torque, depending on normal loading. The test ends when the sample does not fail after 2000000 cycles, or when it has reached a predetermined number of cycles. Test fluid medium used was NaCl mixed with distilled water, a favourable environment for appearance of fretting corrosion. After the failure of stem fixation at 2450000 cycles, the mantle of bone cement remaining adherent on femoral stem was removed. Microscopic inspection of the femoral stem and of the inner part of the polymetylmethacrilate mantle demonstrated the existence of corrosion of the femoral stem surface beneath the cement mantle, and Fe2O3 deposits on the femoral stem surface and on the inner part of the mantle.

Steady-state acceptor fluorescence anisotropy imaging under evanescent excitation for visualisation of FRET at the plasma membrane.

Directory of Open Access Journals (Sweden)

Viviane Devauges

Full Text Available We present a novel imaging system combining total internal reflection fluorescence (TIRF microscopy with measurement of steady-state acceptor fluorescence anisotropy in order to perform live cell Förster Resonance Energy Transfer (FRET imaging at the plasma membrane. We compare directly the imaging performance of fluorescence anisotropy resolved TIRF with epifluorescence illumination. The use of high numerical aperture objective for TIRF required correction for induced depolarization factors. This arrangement enabled visualisation of conformational changes of a Raichu-Cdc42 FRET biosensor by measurement of intramolecular FRET between eGFP and mRFP1. Higher activity of the probe was found at the cell plasma membrane compared to intracellularly. Imaging fluorescence anisotropy in TIRF allowed clear differentiation of the Raichu-Cdc42 biosensor from negative control mutants. Finally, inhibition of Cdc42 was imaged dynamically in live cells, where we show temporal changes of the activity of the Raichu-Cdc42 biosensor.

Highly sensitive FRET-based fluorescence immunoassay for aflatoxin B1 using cadmium telluride quantum dots

International Nuclear Information System (INIS)

Zekavati, Roya; Bayat, Mansour; Safi, Shahabeddin; Hashemi, Seyed Jamal; Rahmani-Cherati, Tavoos; Tabatabaei, Meisam; Mohsenifar, Afshin

2013-01-01

We report on a competitive immunoassay for the determination of aflatoxin B1 using fluorescence resonance energy transfer (FRET) from anti-aflatoxin B1 antibody (immobilized on the shell of CdTe quantum dots) to Rhodamine 123 (Rho 123-labeled aflatoxin B1 bound to albumin). The highly specific immuno reaction between the antibody against aflatoxin B1 on the QDs and the labeled-aflatoxin B1 brings the Rho 123 fluorophore (acting as the acceptor) and the QDs (acting as the donor) in close spatial proximity and causes FRET to occur upon photoexcitation of the QDs. In the absence of unlabeled aflatoxin B1, the antigen-antibody complex is stable, and strong emission resulting from the FRET from QDs to labeled aflatoxin B1 is observed. In the presence of aflatoxin B1, it will compete with the labeled aflatoxin B1-albumin complex for binding to the antibody-QDs conjugate so that FRET will be increasingly suppressed. The reduction in the fluorescence intensity of the acceptor correlates well with the concentration of aflatoxin B1. The feasibility of the method was established by the detection of aflatoxin B1 in spiked human serum. There is a linear relationship between the increased fluorescence intensity of Rho 123 with increasing concentration of aflatoxin B1 in spike human serum, over the range of 0.1–0.6 μmol·mL −1 . The limit of detection is 2 × 10 −11 M. This homogeneous competitive detection scheme is simple, rapid and efficient, and does not require excessive washing and separation steps. (author)

Three-color confocal Förster (or fluorescence) resonance energy transfer microscopy: Quantitative analysis of protein interactions in the nucleation of actin filaments in live cells.

Science.gov (United States)

Wallrabe, Horst; Sun, Yuansheng; Fang, Xiaolan; Periasamy, Ammasi; Bloom, George S

2015-06-01

Experiments using live cell 3-color Förster (or fluorescence) resonance energy transfer (FRET) microscopy and corresponding in vitro biochemical reconstitution of the same proteins were conducted to evaluate actin filament nucleation. A novel application of 3-color FRET data is demonstrated, extending the analysis beyond the customary energy-transfer efficiency (E%) calculations. MDCK cells were transfected for coexpression of Teal-N-WASP/Venus-IQGAP1/mRFP1-Rac1, Teal-N-WASP/Venus-IQGAP1/mRFP1-Cdc42, CFP-Rac1/Venus-IQGAP1/mCherry-actin, or CFP-Cdc42/Venus-IQGAP1/mCherry-actin, and with single-label equivalents for spectral bleedthrough correction. Using confirmed E% as an entry point, fluorescence levels and related ratios were correlated at discrete accumulating levels at cell peripheries. Rising ratios of CFP-Rac1:Venus-IQGAP1 were correlated with lower overall actin fluorescence, whereas the CFP-Cdc42:Venus-IQGAP1 ratio correlated with increased actin fluorescence at low ratios, but was neutral at higher ratios. The new FRET analyses also indicated that rising levels of mRFP1-Cdc42 or mRFP1-Rac1, respectively, promoted or suppressed the association of Teal-N-WASP with Venus-IQGAP1. These 3-color FRET assays further support our in vitro results about the role of IQGAP1, Rac1, and Cdc42 in actin nucleation, and the differential impact of Rac1 and Cdc42 on the association of N-WASP with IQGAP1. In addition, this study emphasizes the power of 3-color FRET as a systems biology strategy for simultaneous evaluation of multiple interacting proteins in individual live cells. © 2015 International Society for Advancement of Cytometry.

SH2 Domain-Based FRET Biosensor for Measuring BCR-ABL Activity in Living CML Cells.

Science.gov (United States)

Fujioka, Mari; Asano, Yumi; Nakada, Shigeyuki; Ohba, Yusuke

2017-01-01

Fluorescent proteins (FPs) displaying distinct spectra have shed their light on a wide range of biological functions. Moreover, sophisticated biosensors engineered to contain single or multiple FPs, including Förster resonance energy transfer (FRET)-based biosensors, spatiotemporally reveal the molecular mechanisms underlying a variety of pathophysiological processes. However, their usefulness for applied life sciences has yet to be fully explored. Recently, our research group has begun to expand the potential of FPs from basic biological research to the clinic. Here, we describe a method to evaluate the responsiveness of leukemia cells from patients to tyrosine kinase inhibitors using a biosensor based on FP technology and the principle of FRET. Upon phosphorylation of the tyrosine residue of the biosensor, binding of the SH2 domain to phosphotyrosine induces conformational change of the biosensor and brings the donor and acceptor FPs into close proximity. Therefore, kinase activity and response to kinase inhibitors can be monitored by an increase and a decrease in FRET efficiency, respectively. As in basic research, this biosensor resolves hitherto arduous tasks and may provide innovative technological advances in clinical laboratory examinations. State-of-the-art detection devices that enable such innovation are also introduced.

Critical Shell Thickness of Core/Shell Upconversion Luminescence Nanoplatform for FRET Application

NARCIS (Netherlands)

Wang, Yu; Liu, Kai; Liu, Xiaomin; Dohnalova, Katerina; Gregorkiewicz, Tom; Kong, Xianggui; Aalders, Maurice C. G.; Buma, Wybren J.; Zhang, Hong

2011-01-01

Almost all the luminescence upconversion nanoparticles used for Forster resonant energy transfer (FRET) applications are bare cores based on the consideration that the energy transfer efficiency is optimized because the distance between energy donors and acceptors is minimized. On the other hand, it

Determination of Equilibrium Constant and Relative Brightness in FRET-FCS by Including the Third-Order Correlations.

Science.gov (United States)

Meng, Lingyi; He, Shanshan; Zhao, Xin Sheng

2017-12-21

Fluorescence correlation spectroscopy (FCS) encodes the information on the equilibrium constant (K), the relative fluorescence brightness of fluorophore (Q), and the forward and backward reaction rate constants (k + and k - ) on a physical or chemical relaxation. However, it has been a long-standing problem to completely resolve the FCS data to get the thermodynamic and kinetic information. Recently, we have solved the problem for fluorescence autocorrelation spectroscopy (FACS). Here, we extend the method to fluorescence cross-correlation spectroscopy (FCCS), which appears when FCS is coupled with fluorescence resonance energy transfer (FRET). Among 12 total second-order and third-order pre-exponential factors in a relaxation process probed by the FRET-FCS technique, 3 are independent. We presented and discussed 3 sets of explicit solutions to use these pre-exponential factors to calculate K and Q. Together with the relaxation time, the acquired K will allow people to obtain k + and k - , so that the goal of deciphering the FRET-FCS data will be fully reached. The theory is verified by extensive computer simulations and tested experimentally on a system of oligonucleotide hybridization.

Influence of thermal oxidation duration on the microstructure and fretting wear behavior of Ti6Al4V alloy

Energy Technology Data Exchange (ETDEWEB)

Wang, Song [State Key Laboratory of Tribology, Tsinghua University, Beijing 100084 (China); Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China); Liao, Zhenhua [Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China); Biomechanics and Biotechnology Lab, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China); Liu, Yuhong [State Key Laboratory of Tribology, Tsinghua University, Beijing 100084 (China); Department of Mechanical Engineering, Tsinghua University, Beijing 100084 (China); Liu, Weiqiang, E-mail: weiqliu@hotmail.com [State Key Laboratory of Tribology, Tsinghua University, Beijing 100084 (China); Biomechanics and Biotechnology Lab, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China)

2015-06-01

Thermal oxidation under water oxidizing atmosphere was performed on Ti6Al4V alloy under different durations from 2 h to 8 h. Surface characterizations were performed using X-ray diffractometery (XRD), scanning electron microscopy (SEM), Raman spectroscopy, nanoindentation and nano scratch testing. Fretting wear behaviors of untreated and oxidized samples were also examined. The formed oxide coating mainly included rutile TiO{sub 2} as well as a little alumina. The weight gain with respect to the oxidation duration obeyed the linear oxidation kinetics law. The growth of oxide grains was in inadequate growth state of incomplete scale coverage from 2nd to 4th hour duration, in normal growth state from 4th to 6th hour duration while in excessive growth state of oxide particle agglomeration and surface roughening from 6th to 8th (or more than 8th) hour duration. The coating thickness increased from 5 μm to 12 μm as oxidation duration increased from 2 h to 8 h. The increase in duration also increased surface roughness and nano hardness as well as adhesion strength of the film/substrate for oxidized samples. The nano hardness value was 10.06 ± 2.15 GPa and the critical load of failure during nano scratch testing was 554.3 ± 6.44 mN for 4 h treated sample. The untreated and oxidized samples showed a same fretting running status and fretting regime with a displacement amplitude of 200 μm while revealing different fretting failure mechanisms. It was mainly abrasive and adhesive wear under ploughing force for untreated sample, while a mix of 3-body abrasion by rolling oxide particles and severe plastic deformation under high contact stress between two ceramic materials for the oxidized samples. The oxide coating was not worn out and improved the fretting wear resistance of titanium alloy. - Highlights: • A thickness of 5–12 μm rutile TiO{sub 2} coating formed under different oxidation durations. • Weight gain with respect to oxidation duration obeyed linear

FRET-mediated pH-responsive dual fluorescent nanoparticles prepared via click chemistry

Science.gov (United States)

Ouadahi, Karima; Sbargoud, Kamal; Allard, Emmanuel; Larpent, Chantal

2012-01-01

Herein, we report an easy preparation of azide-coated polystyrene-based nanoparticles (15 nm in diameter) and their surface functionalization via CuAAC with fluorophores in water. Resultant dual fluorescent nanoparticles coated with dansyl and pH-sensitive fluorescein moieties as the donor/acceptor FRET pair show a ratiometric response to pH upon excitation at a single wavelength.Herein, we report an easy preparation of azide-coated polystyrene-based nanoparticles (15 nm in diameter) and their surface functionalization via CuAAC with fluorophores in water. Resultant dual fluorescent nanoparticles coated with dansyl and pH-sensitive fluorescein moieties as the donor/acceptor FRET pair show a ratiometric response to pH upon excitation at a single wavelength. Electronic supplementary information (ESI) available: Experimental details and figures S1-S16 as mentioned in the text. See DOI: 10.1039/c2nr11413e

Uzawa algorithm to solve elastic and elastic-plastic fretting wear problems within the bipotential framework

Science.gov (United States)

Ning, Po; Feng, Zhi-Qiang; Quintero, Juan Antonio Rojas; Zhou, Yang-Jing; Peng, Lei

2018-03-01

This paper deals with elastic and elastic-plastic fretting problems. The wear gap is taken into account along with the initial contact distance to obtain the Signorini conditions. Both the Signorini conditions and the Coulomb friction laws are written in a compact form. Within the bipotential framework, an augmented Lagrangian method is applied to calculate the contact forces. The Archard wear law is then used to calculate the wear gap at the contact surface. The local fretting problems are solved via the Uzawa algorithm. Numerical examples are performed to show the efficiency and accuracy of the proposed approach. The influence of plasticity has been discussed.

Design specifications to ensure flow-induced vibration and fretting-wear performance in CANDU steam generators and heat exchangers

International Nuclear Information System (INIS)

Janzen, V.P.; Han, Y.; Pettigrew, M.J.

2009-01-01

Preventing flow-induced vibration and fretting-wear problems in steam generators and heat exchangers requires design specifications that bring together specific guidelines, analysis methods, requirements and appropriate performance criteria. This paper outlines the steps required to generate and support such design specifications for CANDU nuclear steam generators and heat exchangers, and relates them to typical steam-generator design features and computer modeling capabilities. It also describes current issues that are driving changes to flow-induced vibration and fretting-wear specifications that can be applied to the design process for component refurbishment, replacement or new designs. These issues include recent experimental or field evidence for new excitation mechanisms, e.g., the possibility of in-plane fluidelastic instability of U-tubes, the demand for longer reactor and component lifetimes, the need for better predictions of dynamic properties and vibration response, e.g., two-phase random-turbulence excitation, and requirements to consider system 'excursions' or abnormal scenarios, e.g., a main steam line break in the case of steam generators. The paper describes steps being taken to resolve these issues. (author)

Dichotomy Boundary at Aeolis Mensae, Mars: Fretted Terrain Developed in a Sedimentary Deposit

Science.gov (United States)

Irwin, R. P., III; Watters, T. R.; Howard, A. D.; Maxwell, T. A.; Craddock, R. A.

2003-03-01

Fretted terrain in Aeolis Mensae, Mars, developed in a sedimentary deposit. A thick, massive unit with a capping layer or duricrust overlies a more durable layered sequence. Wind, collapse, and minor fluvial activity contributed to degradation.

FRET based integrated pyrene-AgNPs system for detection of Hg (II) and pyrene dimer: Applications to environmental analysis

Science.gov (United States)

Walekar, Laxman S.; Hu, Peidong; Vafaei Molamahmood, Hamed; Long, Mingce

2018-06-01

The integrated system of pyrene and cetyltrimethyl ammonium bromide (CTAB) capped silver nanoparticles (AgNPs) with a distance (r) of 2.78 nm has been developed for the detection of Hg (II) and pyrene dimer. The interaction between pyrene and AgNPs results in the fluorescence quenching of pyrene due to the energy transfer, whose mechanism can be attributed to the Forster Resonance Energy Transfer (FRET) supported by experimental observation and theoretical calculations. The developed probe shows a highly selective and sensitive response towards Hg (II) probably due to the amalgam formation, which results in the fluorescence recovery (90%) of pyrene and color change of solution from yellowish brown to colorless. The addition of Hg (II) may increase the distance between pyrene and AgNPs undergoes the 'FRET OFF' process. This system gives a selective response towards Hg (II) over other competing metal ions. Under the optimal condition, the system offers good linearity between 0.1 and 0.6 μg mL-1 with a detection limit of 62 ng mL-1. In addition, the system also provides an effective platform for detection of pyrene in its dimer form even at very low concentrations (10 ng mL-1) on the surface of AgNPs. Therefore, it could be used as effective alternatives for the detection of Hg (II) as well as pyrene simultaneously.

A combined experimental and finite element approach to analyse the fretting mechanism of the head-stem taper junction in total hip replacement

NARCIS (Netherlands)

Bitter, T.; Khan, I.; Marriott, T.; Lovelady, E.; Verdonschot, N.J.; Janssen, D.W.

2017-01-01

Fretting corrosion at the taper interface of modular hip implants has been implicated as a possible cause of implant failure. This study was set up to gain more insight in the taper mechanics that lead to fretting corrosion. The objectives of this study therefore were (1) to select experimental

Correlative FRET: new method improves rigor and reproducibility in determining distances within synaptic nanoscale architecture

Science.gov (United States)

Shinogle-Decker, Heather; Martinez-Rivera, Noraida; O'Brien, John; Powell, Richard D.; Joshi, Vishwas N.; Connell, Samuel; Rosa-Molinar, Eduardo

2018-02-01

A new correlative Förster Resonance Energy Transfer (FRET) microscopy method using FluoroNanogold™, a fluorescent immunoprobe with a covalently attached Nanogold® particle (1.4nm Au), overcomes resolution limitations in determining distances within synaptic nanoscale architecture. FRET by acceptor photobleaching has long been used as a method to increase fluorescence resolution. The transfer of energy from a donor to an acceptor generally occurs between 10-100Å, which is the relative distance between the donor molecule and the acceptor molecule. For the correlative FRET microscopy method using FluoroNanogold™, we immuno-labeled GFP-tagged-HeLa-expressing Connexin 35 (Cx35) with anti-GFP and with anti-Cx35/36 antibodies, and then photo-bleached the Cx before processing the sample for electron microscopic imaging. Preliminary studies reveal the use of Alexa Fluor® 594 FluoroNanogold™ slightly increases FRET distance to 70Å, in contrast to the 62.5Å using AlexaFluor 594®. Preliminary studies also show that using a FluoroNanogold™ probe inhibits photobleaching. After one photobleaching session, Alexa Fluor 594® fluorescence dropped to 19% of its original fluorescence; in contrast, after one photobleaching session, Alexa Fluor 594® FluoroNanogold™ fluorescence dropped to 53% of its original intensity. This result confirms that Alexa Fluor 594® FluoroNanogold™ is a much better donor probe than is Alexa Fluor 594®. The new method (a) creates a double confirmation method in determining structure and orientation of synaptic architecture, (b) allows development of a two-dimensional in vitro model to be used for precise testing of multiple parameters, and (c) increases throughput. Future work will include development of FluoroNanogold™ probes with different sizes of gold for additional correlative microscopy studies.

A Study on Corrosion and Fretting Wear Resistance of Alloy 690 Tubes

Energy Technology Data Exchange (ETDEWEB)

Won, Ju Jin; Min, Su Jung; Kim, Myeong Su; Kim, Kyu Tae [Dongguk Univ., Gyeongju (Korea, Republic of)

2013-10-15

In this article, the effects of such failures have on the materials of alloy 690 are assessed. The corroded volume variation and mass decreased continuously with time. However, the oxide volume changes in an irregular pattern since the oxide formed on the alloy 690 metal may be detached due to the flake formation. The amount of the fretting wear increased with time. It can be seen that the wear rate increased with time and reduced at the later time. The test results show that the ductility decreased as corrosion increases. Alloy 690 is broadly used as a material of nuclear power plant's steam generator tubes because of its excellent mechanical strength, corrosion properties, wear properties and stability at a high temperature. However, the tubes for nuclear power plant's steam generators become a major threat for lifetime management and efficient operation of nuclear power plant due to various corrosion and fretting wear failures caused by flow-induced vibration (FIV) that occurs between tubes.

Medical diagnosis and remote sensing at fiber-tip: picosecond resolved FRET sensor

Science.gov (United States)

Polley, Nabarun; Pal, Samir Kumar

2016-03-01

Förster Resonance Energy Transfer (FRET) strategy in popular in fiber-optic sensing. However, the steady state emission quenching of the donor is inadequate to conclude FRET. The resonance type energy transfer from one molecule (donor) to other (acceptor) should meet few key properties including donor to acceptor energy migration in non-radiative way. In the present study, we have coupled the evanescent field of an optical fiber to the covalently attached donor (dansyl) molecules at the fiber tip. By using picosecond resolved time correlated single photon counting (TCSPC) we have demonstrated that dansyl at the fiber tip transfers energy to a well known DNA-intercalating dye ethidium. Our ultrafast detection scheme selectively distinguishes the probe (dansyl) emission from the intrinsic emission of the fiber. We have also used the setup for the remote sensing of the dielectric constant (polarity) of an environment. We have finally implemented the detection mechanism to detect an industrial synthetic dye methylene blue (MB) in water.

Accurate Quantitative Sensing of Intracellular pH based on Self-ratiometric Upconversion Luminescent Nanoprobe.

Science.gov (United States)

Li, Cuixia; Zuo, Jing; Zhang, Li; Chang, Yulei; Zhang, Youlin; Tu, Langping; Liu, Xiaomin; Xue, Bin; Li, Qiqing; Zhao, Huiying; Zhang, Hong; Kong, Xianggui

2016-12-09

Accurate quantitation of intracellular pH (pH i ) is of great importance in revealing the cellular activities and early warning of diseases. A series of fluorescence-based nano-bioprobes composed of different nanoparticles or/and dye pairs have already been developed for pH i sensing. Till now, biological auto-fluorescence background upon UV-Vis excitation and severe photo-bleaching of dyes are the two main factors impeding the accurate quantitative detection of pH i . Herein, we have developed a self-ratiometric luminescence nanoprobe based on förster resonant energy transfer (FRET) for probing pH i , in which pH-sensitive fluorescein isothiocyanate (FITC) and upconversion nanoparticles (UCNPs) were served as energy acceptor and donor, respectively. Under 980 nm excitation, upconversion emission bands at 475 nm and 645 nm of NaYF 4 :Yb 3+ , Tm 3+ UCNPs were used as pH i response and self-ratiometric reference signal, respectively. This direct quantitative sensing approach has circumvented the traditional software-based subsequent processing of images which may lead to relatively large uncertainty of the results. Due to efficient FRET and fluorescence background free, a highly-sensitive and accurate sensing has been achieved, featured by 3.56 per unit change in pH i value 3.0-7.0 with deviation less than 0.43. This approach shall facilitate the researches in pH i related areas and development of the intracellular drug delivery systems.

Abscisic acid dynamics in roots detected with genetically encoded FRET sensors

Science.gov (United States)

Jones, Alexander M; Danielson, Jonas ÅH; ManojKumar, Shruti N; Lanquar, Viviane; Grossmann, Guido; Frommer, Wolf B

2014-01-01

Cytosolic hormone levels must be tightly controlled at the level of influx, efflux, synthesis, degradation and compartmentation. To determine ABA dynamics at the single cell level, FRET sensors (ABACUS) covering a range ∼0.2–800 µM were engineered using structure-guided design and a high-throughput screening platform. When expressed in yeast, ABACUS1 detected concentrative ABA uptake mediated by the AIT1/NRT1.2 transporter. Arabidopsis roots expressing ABACUS1-2µ (Kd∼2 µM) and ABACUS1-80µ (Kd∼80 µM) respond to perfusion with ABA in a concentration-dependent manner. The properties of the observed ABA accumulation in roots appear incompatible with the activity of known ABA transporters (AIT1, ABCG40). ABACUS reveals effects of external ABA on homeostasis, that is, ABA-triggered induction of ABA degradation, modification, or compartmentation. ABACUS can be used to study ABA responses in mutants and quantitatively monitor ABA translocation and regulation, and identify missing components. The sensor screening platform promises to enable rapid fine-tuning of the ABA sensors and engineering of plant and animal hormone sensors to advance our understanding of hormone signaling. DOI: http://dx.doi.org/10.7554/eLife.01741.001 PMID:24737862

Förster Resonance Energy Transfer (FRET as a Tool for Dissecting the Molecular Mechanisms for Maturation of the Shigella Type III Secretion Needle Tip Complex

Directory of Open Access Journals (Sweden)

William D. Picking

2012-11-01

Full Text Available Förster resonance energy transfer (FRET provides a powerful tool for monitoring intermolecular interactions and a sensitive technique for studying Å-level protein conformational changes. One system that has particularly benefited from the sensitivity and diversity of FRET measurements is the maturation of the Shigella type III secretion apparatus (T3SA needle tip complex. The Shigella T3SA delivers effector proteins into intestinal cells to promote bacterial invasion and spread. The T3SA is comprised of a basal body that spans the bacterial envelope and a needle with an exposed tip complex that matures in response to environmental stimuli. FRET measurements demonstrated bile salt binding by the nascent needle tip protein IpaD and also mapped resulting structural changes which led to the recruitment of the translocator IpaB. At the needle tip IpaB acts as a sensor for host cell contact but prior to secretion, it is stored as a heterodimeric complex with the chaperone IpgC. FRET analyses showed that chaperone binding to IpaB’s N-terminal domain causes a conformational change in the latter. These FRET analyses, with other biophysical methods, have been central to understanding T3SA maturation and will be highlighted, focusing on the details of the FRET measurements and the relevance to this particular system.

Probing the Conformational Landscape of DNA Polymerases Using Diffusion-Based Single-Molecule FRET

NARCIS (Netherlands)

Hohlbein, J.; Kapanidis, A.N.

2016-01-01

Monitoring conformational changes in DNA polymerases using single-molecule Förster resonance energy transfer (smFRET) has provided new tools for studying fidelity-related mechanisms that promote the rejection of incorrect nucleotides before DNA synthesis. In addition to the previously known open

Combination and Integration of Qualitative and Quantitative Analysis

Directory of Open Access Journals (Sweden)

Philipp Mayring

2001-02-01

Full Text Available In this paper, I am going to outline ways of combining qualitative and quantitative steps of analysis on five levels. On the technical level, programs for the computer-aided analysis of qualitative data offer various combinations. Where the data are concerned, the employment of categories (for instance by using qualitative content analysis allows for combining qualitative and quantitative forms of data analysis. On the individual level, the creation of types and the inductive generalisation of cases allow for proceeding from individual case material to quantitative generalisations. As for research design, different models can be distinguished (preliminary study, generalisation, elaboration, triangulation which combine qualitative and quantitative steps of analysis. Where the logic of research is concerned, it can be shown that an extended process model which combined qualitative and quantitative research can be appropriate and thus lead to an integration of the two approaches. URN: urn:nbn:de:0114-fqs010162

The Use of Two-Photon FRET-FLIM to Study Protein Interactions During Nuclear Envelope Fusion In Vivo and In Vitro.

Science.gov (United States)

Byrne, Richard D; Larijani, Banafshé; Poccia, Dominic L

2016-01-01

FRET-FLIM techniques have wide application in the study of protein and protein-lipid interactions in cells. We have pioneered an imaging platform for accurate detection of functional states of proteins and their interactions in fixed cells. This platform, two-site-amplified Förster resonance energy transfer (a-FRET), allows greater signal generation while retaining minimal noise thus enabling application of fluorescence lifetime imaging microscopy (FLIM) to be routinely deployed in different types of cells and tissue. We have used the method described here, time-resolved FRET monitored by two-photon FLIM, to demonstrate the direct interaction of Phospholipase Cγ (PLCγ) by Src Family Kinase 1 (SFK1) during nuclear envelope formation and during male and female pronuclear membrane fusion in fertilized sea urchin eggs. We describe here a generic method that can be applied to monitor any proteins of interest.

Energy Dispersive Spectrometry and Quantitative Analysis Short Course. Introduction to X-ray Energy Dispersive Spectrometry and Quantitative Analysis

Science.gov (United States)

Carpenter, Paul; Curreri, Peter A. (Technical Monitor)

2002-01-01

This course will cover practical applications of the energy-dispersive spectrometer (EDS) to x-ray microanalysis. Topics covered will include detector technology, advances in pulse processing, resolution and performance monitoring, detector modeling, peak deconvolution and fitting, qualitative and quantitative analysis, compositional mapping, and standards. An emphasis will be placed on use of the EDS for quantitative analysis, with discussion of typical problems encountered in the analysis of a wide range of materials and sample geometries.

Electrophoresis- and FRET-Based Measures of Serpin Polymerization.

Science.gov (United States)

Faull, Sarah V; Brown, Anwen E; Haq, Imran; Irving, James A

2017-01-01

Many serpinopathies, including alpha-1 antitrypsin (A1AT) deficiency, are associated with the formation of unbranched polymer chains of mutant serpins. In vivo, this deficiency is the result of mutations that cause kinetic or thermodynamic destabilization of the molecule. However, polymerization can also be induced in vitro from mutant or wild-type serpins under destabilizing conditions. The characteristics of the resulting polymers are dependent upon induction conditions. Due to their relationship to disease, serpin polymers, mainly those formed from A1AT, have been widely studied. Here, we describe Förster resonance energy transfer (FRET) and gel-based approaches for their characterization.

Effects of Plasma ZrN Metallurgy and Shot Peening Duplex Treatment on Fretting Wear and Fretting Fatigue Behavior of Ti6Al4V Alloy.

Science.gov (United States)

Tang, Jingang; Liu, Daoxin; Zhang, Xiaohua; Du, Dongxing; Yu, Shouming

2016-03-23

A metallurgical zirconium nitride (ZrN) layer was fabricated using glow metallurgy using nitriding with zirconiuming prior treatment of the Ti6Al4V alloy. The microstructure, composition and microhardness of the corresponding layer were studied. The influence of this treatment on fretting wear (FW) and fretting fatigue (FF) behavior of the Ti6Al4V alloy was studied. The composite layer consisted of an 8-μm-thick ZrN compound layer and a 50-μm-thick nitrogen-rich Zr-Ti solid solution layer. The surface microhardness of the composite layer is 1775 HK 0.1 . A gradient in cross-sectional microhardness distribution exists in the layer. The plasma ZrN metallurgical layer improves the FW resistance of the Ti6Al4V alloy, but reduces the base FF resistance. This occurs because the improvement in surface hardness results in lowering of the toughness and increasing in the notch sensitivity. Compared with shot peening treatment, plasma ZrN metallurgy and shot peening composite treatment improves the FW resistance and enhances the FF resistance of the Ti6Al4V alloy. This is attributed to the introduction of a compressive stress field. The combination of toughness, strength, FW resistance and fatigue resistance enhance the FF resistance for titanium alloy.

Inspection, visualisation and analysis of quantitative proteomics data

OpenAIRE

Gatto, Laurent

2016-01-01

Material Quantitative Proteomics and Data Analysis Course. 4 - 5 April 2016, Queen Hotel, Chester, UK Table D - Inspection, visualisation and analysis of quantitative proteomics data, Laurent Gatto (University of Cambridge)

FRET-based biosensors for the detection and quantification of AI-2 class of quorum sensing compounds.

Science.gov (United States)

Rajamani, Sathish; Sayre, Richard

2011-01-01

Intercellular small molecular weight signaling molecules modulate a variety of biological functions in bacteria. One of the more complex behaviors mediated by intercellular signaling molecules is the suite of activities regulated by quorum sensing molecules. These molecules mediate a variety of population-dependent responses, including the expression of genes that regulate bioluminescence, type III secretion, siderophore production, colony morphology, biofilm formation, and metalloprotease production. Given their central role in regulating these responses, the detection and quantification of QS molecules has important practical implications. Until recently, the detection of QS molecules from Gram-negative bacteria has relied primarily on bacterial reporter systems. These bioassays though immensely useful are subject to interference by compounds that affect bacterial growth and metabolism. In addition, the reporter response is highly dependent on culture age and cell population density. To overcome such limitations, we developed an in vitro protein-based assay system for the rapid detection and quantification of the furanosyl borate diester (BAI-2) subclass of autoinducer-2 (AI-2) QS molecules. The biosensor is based on the interaction of BAI-2 with the Vibrio harveyi QS receptor LuxP. Conformation changes associated with BAI-2 binding to the LuxP receptor change the orientation of cyan and yellow variants of GFP (CFP and YFP) fused the N- and C-termini, respectively, of the LuxP receptor. LuxP-BAI2 binding induces changes in fluorescence resonance energy transfer (FRET) between CFP and YFP, whose magnitude of change is ligand concentration dependent. A set of ligand-insensitive LuxP-mutant FRET protein sensor was also developed for use as control biosensors. The FRET-based BAI-2 biosensor responds selectively to both synthetic and biologically derived BAI-2compounds. This report describes the use of the LuxP-FRET biosensor for the detection and quantification of

Experimental Simulation of Flow-Induced Vibration for Developing a Grid-to-Rod Fretting Model

Energy Technology Data Exchange (ETDEWEB)

Lee, Youngho; Kim, Hyungkyu; Kang, Heungseok [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of)

2013-05-15

GTRF margin was calculated based on the fuel reliabilities program of operating power plants. But they have not accumulated sufficient experience under challenging operating conditions to be considered proven solutions. In addition, GTRF behaviors were significantly differed according to the plant types, operating condition and fuel types. So, analytical methods to resolve GTRF degradations are considered as difficult procedures for actual application. One of the most important problems is that it is difficult to evaluate the GTRF resistance of new spacer grid under operating power plant condition. Up to now, as a consequence, compliance with the fretting wear limit (typically 10% of the cladding thickness) is checked a posteriori, through post-irradiation examination. Therefore, in this study, rod simulation method for determining GTRF resistance of new spacer grid was proposed with a specially designed wear tester. This simulator enables us to examine the spacer grid shape effect under relatively short development period. In addition, for developing GTRF model, flow-induced vibration (FIV) was measured with different major variables such as GTR clearance, flow rate, etc. Fretting wear tests of nuclear fuel rods (i. e. grid-to-rod fretting) have been performed to examine the flow rate effect by using a specially designed test section with a simulated primary coolant. Based on above results, developed FIV-wear simulator could be effective to examine the spacer grid shape effect with short development period. Further study will be discussed on the GTR clearance effect with various spacer grid shapes.

Tumor specific lung cancer diagnostics with multiplexed FRET immunoassays

Science.gov (United States)

Geißler, D.; Hill, D.; Löhmannsröben, H.-G.; Thomas, E.; Lavigne, A.; Darbouret, B.; Bois, E.; Charbonnière, L. J.; Ziessel, R. F.; Hildebrandt, N.

2010-02-01

An optical multiplexed homogeneous (liquid phase) immunoassay based on FRET from a terbium complex to eight different fluorescent dyes is presented. We achieved highly sensitive parallel detection of four different lung cancer specific tumor markers (CEA, NSE, SCC and CYFRA21-1) within a single assay and show a proof-of-principle for 5- fold multiplexing. The method is well suited for fast and low-cost miniaturized point-of-care testing as well as for highthroughput screening in a broad range of in-vitro diagnostic applications.

Quantitative Moessbauer analysis

International Nuclear Information System (INIS)

Collins, R.L.

1978-01-01

The quantitative analysis of Moessbauer data, as in the measurement of Fe 3+ /Fe 2+ concentration, has not been possible because of the different mean square velocities (x 2 ) of Moessbauer nuclei at chemically different sites. A method is now described which, based on Moessbauer data at several temperatures, permits the comparison of absorption areas at (x 2 )=0. (Auth.)

Quantitative Data Analysis--In the Graduate Curriculum

Science.gov (United States)

Albers, Michael J.

2017-01-01

A quantitative research study collects numerical data that must be analyzed to help draw the study's conclusions. Teaching quantitative data analysis is not teaching number crunching, but teaching a way of critical thinking for how to analyze the data. The goal of data analysis is to reveal the underlying patterns, trends, and relationships of a…

A New FRET-Based Sensitive DNA Sensor for Medical Diagnostics using PNA Probe and Water-Soluble Blue Light Emitting Polymer

Directory of Open Access Journals (Sweden)

Nidhi Mathur

2008-01-01

Full Text Available A reliable, fast, and low-cost biosensor for medical diagnostics using DNA sequence detection has been developed and tested for the detection of the bacterium “Bacillus anthracis.” In this sensor, Poly [9,9-di (6,6′- N, N′ trimethylammonium hexylfluorenyl-2, 7-diyl-alt-co- (1,4-phenylene] dibromide salt (PFP has been taken as cationic conjugated polymer (CCP and PNA attached with fluorescein dye (PNAC∗ as a probe. The basic principle of this sensor is that when a PNAC∗ probe is hybridized with a single strand DNA (ssDNA having complementary sequence, Forster resonance energy transfer (FRET may take place from PFP to the PNAC∗/DNA complex. If the FRET is efficient, the photoluminescence from the PFP will be highly quenched and that from PNAC∗ will be enhanced. On the other hand, if the DNA sequence is noncomplementary to PNA, FRET will not occur.

Intramolecular ex vivo Fluorescence Resonance Energy Transfer (FRET of Dihydropyridine Receptor (DHPR β1a Subunit Reveals Conformational Change Induced by RYR1 in Mouse Skeletal Myotubes.

Directory of Open Access Journals (Sweden)

Dipankar Bhattacharya

Full Text Available The dihydropyridine receptor (DHPR β1a subunit is essential for skeletal muscle excitation-contraction coupling, but the structural organization of β1a as part of the macromolecular DHPR-ryanodine receptor type I (RyR1 complex is still debatable. We used fluorescence resonance energy transfer (FRET to probe proximity relationships within the β1a subunit in cultured skeletal myotubes lacking or expressing RyR1. The fluorescein biarsenical reagent FlAsH was used as the FRET acceptor, which exhibits fluorescence upon binding to specific tetracysteine motifs, and enhanced cyan fluorescent protein (CFP was used as the FRET donor. Ten β1a reporter constructs were generated by inserting the CCPGCC FlAsH binding motif into five positions probing the five domains of β1a with either carboxyl or amino terminal fused CFP. FRET efficiency was largest when CCPGCC was positioned next to CFP, and significant intramolecular FRET was observed for all constructs suggesting that in situ the β1a subunit has a relatively compact conformation in which the carboxyl and amino termini are not extended. Comparison of the FRET efficiency in wild type to that in dyspedic (lacking RyR1 myotubes revealed that in only one construct (H458 CCPGCC β1a -CFP FRET efficiency was specifically altered by the presence of RyR1. The present study reveals that the C-terminal of the β1a subunit changes conformation in the presence of RyR1 consistent with an interaction between the C-terminal of β1a and RyR1 in resting myotubes.

Original methods of quantitative analysis developed for diverse samples in various research fields. Quantitative analysis at NMCC

International Nuclear Information System (INIS)

Sera, Koichiro

2003-01-01

Nishina Memorial Cyclotron Center (NMCC) has been opened for nationwide-common utilization of positron nuclear medicine (PET) and PIXE since April 1993. At the present time, nearly 40 subjects of PIXE in various research fields are pursued here, and more than 50,000 samples have been analyzed up to the present. In order to perform quantitative analyses of diverse samples, technical developments in sample preparation, measurement and data analysis have been continuously carried out. Especially, a standard-free method for quantitative analysis'' made it possible to perform analysis of infinitesimal samples, powdered samples and untreated bio samples, which could not be well analyzed quantitatively in the past. The standard-free method'' and a ''powdered internal standard method'' made the process for target preparation quite easier. It has been confirmed that results obtained by these methods show satisfactory accuracy and reproducibility preventing any ambiguity coming from complicated target preparation processes. (author)

A fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) for the detection of mecA gene sequence of Staphylococcus aureus.

Science.gov (United States)

Shi, Jingyu; Chan, Chunyu; Pang, Yukting; Ye, Weiwei; Tian, Feng; Lyu, Jing; Zhang, Yu; Yang, Mo

2015-05-15

In this work, a novel fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) pairs was developed for Staphylococcus aureus specific gene sequence detection. This FRET biosensor platform was realized by immobilization of capture probes on GQDs and conjugation of reporter probes on AuNPs. Target oligos then co-hybridized with capture probes and reporter probes to form a sandwich structure which brought GQDs and AuNPs to close proximity to trigger FRET effect. The fluorescence signals before and after addition of targets were measured and the fluorescence quenching efficiency could reach around 87% with 100 nM target oligo. The limit of detection (LOD) of this FRET biosensor was around 1 nM for S.aureus gene detection. Experiments with both single-base mismatched oligos and double-base mismatched oligos demonstrated the good sequence selectivity of this FRET biosensor. Copyright © 2014 Elsevier B.V. All rights reserved.

Fretting wear of ZrN and Zr(21% Hf)N coatings

Energy Technology Data Exchange (ETDEWEB)

Atar, E. [Gebze Inst. of Tech., Material Science and Engineering Dept., Kocaeli (Turkey); Cimenoglu, H.; Kayali, E.S. [Istanbul Technical Univ., Dept. of Metallurgy and Materials Engineering, Istanbul (Turkey)

2004-07-01

In this study, the wear behaviours of ZrN and Zr(21% Hf)N coatings, deposited on hardened AISI D2 cold work tool steel were examined by a fretting wear tester. The hardness of ZrN and Zr(21% Hf)N coatings were almost the same, where as they exhibited different wear resistance. Addition of 21% Hf to ZrN coating achieved about 25% increase in the wear resistance. (orig.)

Fretting wear of ZrN and Zr(21% Hf)N coatings

International Nuclear Information System (INIS)

Atar, E.; Cimenoglu, H.; Kayali, E.S.

2004-01-01

In this study, the wear behaviours of ZrN and Zr(21% Hf)N coatings, deposited on hardened AISI D2 cold work tool steel were examined by a fretting wear tester. The hardness of ZrN and Zr(21% Hf)N coatings were almost the same, where as they exhibited different wear resistance. Addition of 21% Hf to ZrN coating achieved about 25% increase in the wear resistance. (orig.)

A combined experimental and finite element approach to analyse the fretting mechanism of the head-stem taper junction in total hip replacement.

Science.gov (United States)

Bitter, Thom; Khan, Imran; Marriott, Tim; Lovelady, Elaine; Verdonschot, Nico; Janssen, Dennis

2017-09-01

Fretting corrosion at the taper interface of modular hip implants has been implicated as a possible cause of implant failure. This study was set up to gain more insight in the taper mechanics that lead to fretting corrosion. The objectives of this study therefore were (1) to select experimental loading conditions to reproduce clinically relevant fretting corrosion features observed in retrieved components, (2) to develop a finite element model consistent with the fretting experiments and (3) to apply more complicated loading conditions of activities of daily living to the finite element model to study the taper mechanics. The experiments showed similar wear patterns on the taper surface as observed in retrievals. The finite element wear score based on Archard's law did not correlate well with the amount of material loss measured in the experiments. However, similar patterns were observed between the simulated micromotions and the experimental wear measurements. Although the finite element model could not be validated, the loading conditions based on activities of daily living demonstrate the importance of assembly load on the wear potential. These findings suggest that finite element models that do not incorporate geometry updates to account for wear loss may not be appropriate to predict wear volumes of taper connections.

Quantitative analysis of untreated bio-samples

International Nuclear Information System (INIS)

Sera, K.; Futatsugawa, S.; Matsuda, K.

1999-01-01

A standard-free method of quantitative analysis for untreated samples has been developed. For hair samples, measurements were performed by irradiating with a proton beam a few hairs as they are, and quantitative analysis was carried out by means of a standard-free method developed by ourselves. First, quantitative values of concentration of zinc were derived, then concentration of other elements was obtained by regarding zinc as an internal standard. As the result, values of concentration of sulphur for 40 samples agree well with the average value for a typical Japanese and also with each other within 20%, and validity of the present method could be confirmed. Accuracy was confirmed by comparing the results with those obtained by the usual internal standard method, too. For the purpose of a surface analysis of a bone sample, a very small incidence angle of the proton beam was used, so that both energy loss of the projectile and self-absorption of X-rays become negligible. As the result, consistent values of concentration for many elements were obtained by the standard-free method

FRET-based localization of fluorescent protein insertions within the ryanodine receptor type 1.

Directory of Open Access Journals (Sweden)

Shweta A Raina

Full Text Available Fluorescent protein (FP insertions have often been used to localize primary structure elements in mid-resolution 3D cryo electron microscopic (EM maps of large protein complexes. However, little is known as to the precise spatial relationship between the location of the fused FP and its insertion site within a larger protein. To gain insights into these structural considerations, Förster resonance energy transfer (FRET measurements were used to localize green fluorescent protein (GFP insertions within the ryanodine receptor type 1 (RyR1, a large intracellular Ca(2+ release channel that plays a key role in skeletal muscle excitation contraction coupling. A series of full-length His-tagged GFP-RyR1 fusion constructs were created, expressed in human embryonic kidney (HEK-293T cells and then complexed with Cy3NTA, a His-tag specific FRET acceptor. FRET efficiency values measured from each GFP donor to Cy3NTA bound to each His tag acceptor site were converted into intermolecular distances and the positions of each inserted GFP were then triangulated relative to a previously published X-ray crystal structure of a 559 amino acid RyR1 fragment. We observed that the chromophoric centers of fluorescent proteins inserted into RyR1 can be located as far as 45 Å from their insertion sites and that the fused proteins can also be located in internal cavities within RyR1. These findings should prove useful in interpreting structural results obtained in cryo EM maps using fusions of small fluorescent proteins. More accurate point-to-point distance information may be obtained using complementary orthogonal labeling systems that rely on fluorescent probes that bind directly to amino acid side chains.

Quantitative analysis of boron by neutron radiography

International Nuclear Information System (INIS)

Bayuelken, A.; Boeck, H.; Schachner, H.; Buchberger, T.

1990-01-01

The quantitative determination of boron in ores is a long process with chemical analysis techniques. As nuclear techniques like X-ray fluorescence and activation analysis are not applicable for boron, only the neutron radiography technique, using the high neutron absorption cross section of this element, can be applied for quantitative determinations. This paper describes preliminary tests and calibration experiments carried out at a 250 kW TRIGA reactor. (orig.) [de

Prévision de l'épaisseur du film passif d'un acier inoxydable 316L soumis au fretting corrosion grâce au Point Defect Model, PDM Predicting the steady state thickness of passive films with the Point Defect Model in fretting corrosion experiments

Directory of Open Access Journals (Sweden)

Geringer Jean

2013-11-01

Full Text Available Les implants orthopédiques de hanche ont une durée de vie d'environ 15 ans. Par exemple, la tige fémorale d'un tel implant peut être réalisée en acier inoxydable 316L ou 316LN. Le fretting corrosion, frottement sous petits déplacements, peut se produire pendant la marche humaine en raison des chargements répétés entre le métal de la prothèse et l'os. Plusieurs investigations expérimentales du fretting corrosion ont été entreprises. Cette couche passive de quelques nanomètres, à température ambiante, est le point clef sur lequel repose le développement de notre civilisation, selon certains auteurs. Ce travail vise à prédire les épaisseurs de cette couche passive de l'acier inoxydable soumis au fretting corrosion, avec une attention spécifique sur le rôle des protéines. Le modèle utilisé est basé sur le Point Defect Model, PDM (à une échelle microscopique et une amélioration de ce modèle en prenant en compte le processus de frottement sous petits débattements. L'algorithme génétique a été utilisé pour optimiser la convergence du problème. Les résultats les plus importants sont, comme démontré avec les essais expérimentaux, que l'albumine, la protéine étudiée, empêche les dégradations de l'acier inoxydable aux plus faibles concentrations d'ions chlorure ; ensuite, aux plus fortes concentrations de chlorures, un temps d'incubation est nécessaire pour détruire le film passif. Some implants have approximately a lifetime of 15 years. The femoral stem, for example, should be made of 316L/316LN stainless steel. Fretting corrosion, friction under small displacements, should occur during human gait, due to repeated loadings and un-loadings, between stainless steel and bone for instance. Some experimental investigations of fretting corrosion have been practiced. As well known, metallic alloys and especially stainless steels are covered with a passive film that prevents from the corrosion and degradation

Step size of the rotary proton motor in single FoF1-ATP synthase from a thermoalkaliphilic bacterium by DCO-ALEX FRET

Science.gov (United States)

Hammann, Eva; Zappe, Andrea; Keis, Stefanie; Ernst, Stefan; Matthies, Doreen; Meier, Thomas; Cook, Gregory M.; Börsch, Michael

2012-02-01

Thermophilic enzymes operate at high temperatures but show reduced activities at room temperature. They are in general more stable during preparation and, accordingly, are considered to be more rigid in structure. Crystallization is often easier compared to proteins from bacteria growing at ambient temperatures, especially for membrane proteins. The ATP-producing enzyme FoF1-ATP synthase from thermoalkaliphilic Caldalkalibacillus thermarum strain TA2.A1 is driven by a Fo motor consisting of a ring of 13 c-subunits. We applied a single-molecule Förster resonance energy transfer (FRET) approach using duty cycle-optimized alternating laser excitation (DCO-ALEX) to monitor the expected 13-stepped rotary Fo motor at work. New FRET transition histograms were developed to identify the smaller step sizes compared to the 10-stepped Fo motor of the Escherichia coli enzyme. Dwell time analysis revealed the temperature and the LDAO dependence of the Fo motor activity on the single molecule level. Back-and-forth stepping of the Fo motor occurs fast indicating a high flexibility in the membrane part of this thermophilic enzyme.

Unlubricated Gross Slip Fretting Wear of Metallic Plasma Sprayed Coatings for Ti6A14V Surfaces

National Research Council Canada - National Science Library

Hager, Jr., Carl H; Sanders, Jeffrey H; Sharma, Shashi K

2006-01-01

... to simulate cold engine startup. Alternative coatings such as plasma sprayed molybdenum and nickel were also evaluated because of their potential for reducing fretting wear under certain simulated engine conditions...

Identification of feline immunodeficiency virus subtype-B on St. Kitts, West Indies by quantitative PCR.

Science.gov (United States)

Kelly, Patrick J; Stocking, Ruey; Gao, Dongya; Phillips, Nikol; Xu, Chuanling; Kaltenboeck, Bernhard; Wang, Chengming

2011-07-04

Although antibodies to the feline immunodeficiency virus (FIV) have been detected by SNAP assay in cats from St. Kitts, there have been no molecular studies to further confirm the infection and determine the FIV subtypes present. Total nucleic acids were extracted from EDTA whole blood specimens from 35 cats, followed by quantitative fluorescence resonance energy transfer (FRET) PCR under a six-channel LightCycler 2.0 Instrument with Software version 4.1. Four of 11 stray cats (36 %) but none of 24 owned cats were FIV positive by real-time PCR.  High-resolution melting curve analysis indicated that all four positive cats were infected with FIV subtype-B. This is the first molecular characterization of FIV subtypes on St. Kitts and the results confirm the high prevalence of FIV infection in stray cats on the island.

Towards understanding the E. coli PNP binding mechanism and FRET absence between E. coli PNP and formycin A.

Science.gov (United States)

Prokopowicz, Małgorzata; Greń, Bartosz; Cieśla, Joanna; Kierdaszuk, Borys

2017-11-01

The aim of this study is threefold: (1) augmentation of the knowledge of the E. coli PNP binding mechanism; (2) explanation of the previously observed 'lack of FRET' phenomenon and (3) an introduction of the correction (modified method) for FRET efficiency calculation in the PNP-FA complexes. We present fluorescence studies of the two E. coli PNP mutants (F159Y and F159A) with formycin A (FA), that indicate that the aromatic amino acid is indispensable in the nucleotide binding, additional hydroxyl group at position 159 probably enhances the strength of binding and that the amino acids pair 159-160 has a great impact on the spectroscopic properties of the enzyme. The experiments were carried out in hepes and phosphate buffers, at pH7 and 8.3. Two methods, a conventional and a modified one, that utilizes the dissociation constant, for calculations of the energy transfer efficiency (E) and the acceptor-to-donor distance (r) between FA and the Tyr (energy donor) were employed. Total difference spectra were calculated for emission spectra (λ ex 280nm, 295nm, 305nm and 313nm) for all studied systems. Time-resolved techniques allowed to conclude the existence of a specific structure formed by amino acids at positions 159 and 160. The results showed an unexpected pattern change of FRET in the mutants, when compared to the wild type enzyme and a probable presence of a structure created between 159 and 160 residue, that might influence the binding efficiency. Additionally, we confirmed the indispensable role of the modification of the FRET efficiency (E) calculation on the fraction of enzyme saturation in PNP-FA systems. Copyright © 2017 Elsevier B.V. All rights reserved.

Quantitative image analysis of synovial tissue

NARCIS (Netherlands)

van der Hall, Pascal O.; Kraan, Maarten C.; Tak, Paul Peter

2007-01-01

Quantitative image analysis is a form of imaging that includes microscopic histological quantification, video microscopy, image analysis, and image processing. Hallmarks are the generation of reliable, reproducible, and efficient measurements via strict calibration and step-by-step control of the

High Content Analysis of Compositional Heterogeneities to Study GPCR Oligomerization

DEFF Research Database (Denmark)

Walsh, Samuel McEwen

In this thesis I demonstrate how the natural compositional heterogeneities of synthetic and living cell model systems can be used to quantitate the mechanics of G-protein coupled receptor (GPCR) oligomerization with Förster resonance energy transfer (FRET). The thesis is structured around three a...

Fretting and wear behaviors of Ni/nano-WC composite coatings in dry and wet conditions

International Nuclear Information System (INIS)

Benea, Lidia; Başa, Sorin-Bogdan; Dănăilă, Eliza; Caron, Nadège; Raquet, Olivier; Ponthiaux, Pierre; Celis, Jean-Pierre

2015-01-01

Highlights: • The friction and wear properties of Ni/nano-WC composite were studied. • Nano-WC reinforcement decreased friction coefficient in dry and wet conditions. • Nano-WC reinforcement fraction was seen to be 12 wt.%. • Nanohardness increased by 27% compared to nickel without WC reinforcements. • Ennoblement of OCP corresponding to the Ni/nano-WC composite coating. - Abstract: The fretting and wear behaviors of Ni/nano-WC composite coatings were studied by considering the effect of fretting frequency of 1 Hz during 10,000 cycles, at different applied loads in dry or wet conditions. The studies were performed on a ball-on-disk tribometer and the results were compared with pure Ni coating. The nanohardness of pure Ni and Ni/nano-WC composite coatings was tested by nanoindentation technique. To evaluate the wet wear (tribocorrosion) behavior the open circuit potential (OCP) was measured before, during and after the fretting tests at room temperature in the solution that simulates the primary water circuit of Pressurized Water Reactors (PWRs). The results show that Ni/nano-WC composite coatings exhibited a low friction coefficient, high nanohardness and wear resistance compared with pure Ni coatings under similar experimental conditions. Ni/nano-WC composite coatings were obtained on stainless steel support by electrochemical codeposition of nano-sized WC particles (diameter size of ∼60 nm) with nickel, from a standard nickel Watts plating bath. The surface morphology and the composition of the coatings were characterized by scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDX) respectively

Quantitative Auger analysis of Nb-Ge superconducting alloys

International Nuclear Information System (INIS)

Buitrago, R.H.

1980-01-01

The feasibility of using Auger electron analysis for quantitative analysis was investigated by studying Nb 3 Ge thin-film Auger data with different approaches. A method base on elemental standards gave consistent quantitative values with reported Nb-Ge data. Alloy sputter yields were also calculated and results were consistent with those for pure elements

Structural changes in the cytoplasmic pore of the Kir1.1 channel during pHi-gating probed by FRET.

Science.gov (United States)

Lee, Jay-Ron; Shieh, Ru-Chi

2009-03-06

Kir1.1 channels are important in maintaining K+ homeostasis in the kidney. Intracellular acidification reversibly closes the Kir1.1 channel and thus decreases K+ secretion. In this study, we used Foster resonance energy transfer (FRET) to determine whether the conformation of the cytoplasmic pore changes in response to intracellular pH (pHi)-gating in Kir1.1 channels fused with enhanced cyan fluorescent protein (ECFP) and enhanced yellow fluorescent protein (EYFP) (ECFP-Kir1.1-EYFP). Because the fluorescence intensities of ECFP and EYFP were affected at pHi pHi-gating occurs in the ECFP-Kir1.1-EYFP construct, we examined the FRET efficiencies of an ECFP-S219R-EYFP mutant, which is completed closed at pHi 7.4 and open at pHi 10.0. FRET efficiency was increased from 25% to 40% when the pHi was decreased from 10.0 to 7.4. These results suggest that the conformation of the cytoplasmic pore in the Kir1.1 channel changes in response to pHi gating such that the N- and C-termini move apart from each other at pHi 7.4, when the channel is open.

[Quantitative data analysis for live imaging of bone.

Science.gov (United States)

Seno, Shigeto

Bone tissue is a hard tissue, it was difficult to observe the interior of the bone tissue alive. With the progress of microscopic technology and fluorescent probe technology in recent years, it becomes possible to observe various activities of various cells forming bone society. On the other hand, the quantitative increase in data and the diversification and complexity of the images makes it difficult to perform quantitative analysis by visual inspection. It has been expected to develop a methodology for processing microscopic images and data analysis. In this article, we introduce the research field of bioimage informatics which is the boundary area of biology and information science, and then outline the basic image processing technology for quantitative analysis of live imaging data of bone.

Efficient FRET-based fuorescent ratiometric chemosensors for Fe3+ and its application in living cells

International Nuclear Information System (INIS)

Wang, Cuicui; Liu, Yaqi; Cheng, Junye; Song, Jianhua; Zhao, Yufen; Ye, Yong

2015-01-01

A series of novel FRET-based fluorescent ratiometric chemosensors (L 1 –L 6 ) were designed and synthesized. Sensor L 2 showed reversible and the best selective recognition toward Fe 3+ over other metal ions with a detection limit of 0.418 ppm, which can meet the selective requirements for practical application. Experiment results showed that the response behavior of L 2 toward Fe 3+ is pH independent in weak acid condition (pH 4.0–6.0). In addition, sensor L 2 was successfully applied for ratiometric visualization of Fe 3+ in living cells. - Highlights: • The detection limit of a new FRET probe for Fe 3+ was 0.418 ppm. • The probe exhibited high selectivity and sensitivity detection to Fe 3+ with a pH span of 4.0–6.0. • The significant changes in color could be used for naked-eye detection • The fluorescence imaging experiment demonstrated its value of practical application

Probing plasma membrane microdomains in cowpea protoplasts using lipidated GFP-fusion proteins and multimode FRET microscopy

NARCIS (Netherlands)

Vermeer, J.E.M.; van Munster, E.B.; Vischer, N.O.; Gadella, T.

2004-01-01

Multimode fluorescence resonance energy transfer (FRET) microscopy was applied to study the plasma membrane organization using different lipidated green fluorescent protein (GFP)-fusion proteins co-expressed in cowpea protoplasts. Cyan fluorescent protein (CFP) was fused to the hyper variable region

Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET)

NARCIS (Netherlands)

Lidke, D.S.; Nagy, P.; Barisas, B.G.; Heintzmann, R.; Post, Janine Nicole; Lidke, K.A.; Clayton, A.H.A.; Arndt-jovin, D.J.; Jovin, T.M.

2003-01-01

We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow

Evaluation of a FRET-peptide substrate to predict virulence in Pseudomonas aeruginosa.

Directory of Open Access Journals (Sweden)

Wendy E Kaman

Full Text Available Pseudomonas aeruginosa produces a number of proteases that are associated with virulence and disease progression. A substrate able to detect P. aeruginosa-specific proteolytic activity could help to rapidly alert clinicians to the virulence potential of individual P. aeruginosa strains. For this purpose we designed a set of P. aeruginosa-specific fluorogenic substrates, comprising fluorescence resonance energy transfer (FRET-labeled peptides, and evaluated their applicability to P. aeruginosa virulence in a range of clinical isolates. A FRET-peptide comprising three glycines (3xGly was found to be specific for the detection of P. aeruginosa proteases. Further screening of 97 P. aeruginosa clinical isolates showed a wide variation in 3xGly cleavage activity. The absence of 3xGly degradation by a lasI knock out strain indicated that 3xGly cleavage by P. aeruginosa could be quorum sensing (QS-related, a hypothesis strengthened by the observation of a strong correlation between 3xGly cleavage, LasA staphylolytic activity and pyocyanin production. Additionally, isolates able to cleave 3xGly were more susceptible to the QS inhibiting antibiotic azithromycin (AZM. In conclusion, we designed and evaluated a 3xGly substrate possibly useful as a simple tool to predict virulence and AZM susceptibility.

Investigation of fretting corrosion of vacuum-chrome-plated vt3-1 titanium alloy in pair with unprotected vt3-1 alloy and 40khnma steel

International Nuclear Information System (INIS)

Rojkh, I.L.; Koltunova, L.N.; Vejtsman, M.G.; Birman, Ya.N.; Skosarev, A.V.; Kogan, I.S.

1978-01-01

The character of destruction of contacting surfaces in the process of fretting corrosion of titanium alloy VT3-1 chromized in vacuum in pair with unprotected alloy VT3-1 and steel 40KhNMA has been studied by scanning electron microscopy, electronography, and recording the surface profile. The specific load was 200 kg/cm 2 , vibration amplitude 50 mkm and frequency 500 Hz. It has been established that pairs unprotected with coating are subjected to intensive fretting corrosion especially when they are made of titanium alloy. For the pair chromized alloy VT3-1 - unprotected alloy VT3-1 no destruction of a chromized surface is observed. Vacuum chromium coating in the pair with steel 40KhNMA reveals similar properties as in pair with a titanium alloy. The surface of a steel sample is destroyed because of fretting corrosion, though the intensity of corrosion is lower than in the case of unprotected pairs. Vacuum chromium coating is recommended for protection of titanium alloy VT3-1 from fretting corrosion in pair with steel 40KhNMA or an alloy VT3-1 especially in those cases when various organic coatings are unsuitable

Quantitative phase analysis by neutron diffraction

Energy Technology Data Exchange (ETDEWEB)

Lee, Chang Hee; Song, Su Ho; Lee, Jin Ho; Shim, Hae Seop [Korea Atomic Energy Research Institute, Taejon (Korea)

1999-06-01

This study is to apply quantitative phase analysis (QPA) by neutron diffraction to the round robin samples provided by the International Union of Crystallography(IUCr). We measured neutron diffraction patterns for mixed samples which have several different weight percentages and their unique characteristic features. Neutron diffraction method has been known to be superior to its complementary methods such as X-ray or Synchrotron, but it is still accepted as highly reliable under limited conditions or samples. Neutron diffraction has strong capability especially on oxides due to its scattering cross-section of the oxygen and it can become a more strong tool for analysis on the industrial materials with this quantitative phase analysis techniques. By doing this study, we hope not only to do one of instrument performance tests on our HRPD but also to improve our ability on the analysis of neutron diffraction data by comparing our QPA results with others from any advanced reactor facilities. 14 refs., 4 figs., 6 tabs. (Author)

FRET Response of a Modified Ribose Receptor Expressed in the Diatom Thalassiosira pseudonana

Energy Technology Data Exchange (ETDEWEB)

Miller, Hanna

2011-08-26

The ability to insert complex proteins into silica has many applications including biosensing. Previous research has demonstrated how to direct proteins to the biosilica of diatoms [1]. Here, we show that a complex fusion protein that includes an enzyme, a bacterial ribose periplasmic binding protein, flanked by fluorescent proteins constituting a FRET pair can remain functional in the frustules of living diatoms. A Sil3 tag is attached to the N-terminal end to localize the fusion protein to frustules of the diatom Thalassiosira pseudonana. When ribose was applied, a larger decrease in FRET response was seen in transformed cells than in untransformed cells. Multiple forms of the expression vector were tested to find the optimal system; specifically, a one-vector system was compared to a two-vector system and the gDNA version of the Sil3 localization tag was compared to the cDNA version. The optimal system was found to be a one-vector system with the genomic version of the Sil3 tag to direct the protein to the frustules. Localization of the enzyme to the frustules was further confirmed through cell fluorescence imaging.

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

Förster resonance energy transfer: Role of diffusion of fluorophore orientation and separation in observed shifts of FRET efficiency.

Directory of Open Access Journals (Sweden)

Bram Wallace

Full Text Available Förster resonance energy transfer (FRET is a widely used single-molecule technique for measuring nanoscale distances from changes in the non-radiative transfer of energy between donor and acceptor fluorophores. For macromolecules and complexes this observed transfer efficiency is used to infer changes in molecular conformation under differing experimental conditions. However, sometimes shifts are observed in the FRET efficiency even when there is strong experimental evidence that the molecular conformational state is unchanged. We investigate ways in which such discrepancies can arise from kinetic effects. We show that significant shifts can arise from the interplay between excitation kinetics, orientation diffusion of fluorophores, separation diffusion of fluorophores, and non-emitting quenching.

Investigation on ionic states of 1,2-Dipalmitoyl-sn-glycero-3-phosphorylcholine (DPPC) using organic laser dyes: A FRET study

Energy Technology Data Exchange (ETDEWEB)

Roy, Arpan Datta; Saha, Jaba; Dey, D.; Bhattacharjee, D.; Hussain, Syed Arshad, E-mail: sa_h153@hotmail.com

2017-05-15

Fluorescence Resonance Energy Transfer (FRET) between two organic dyes Fluorescein and Rhodamine 6G were successfully investigated in aqueous solution in presence and absence of 1,2-Dipalmitoyl-sn-glycero-3-phosphorylcholine (DPPC) at different pH. Spectroscopic studies suggest that both the dyes were present mainly as monomer in solution. FRET occurred from Fluorescein to Rhodamine 6G in solutions. Energy transfer efficiency increases in presence of DPPC and the maximum efficiency was 59.3% when the concentration of DPPC was 1.4×10{sup −4} M at ambient condition. pH plays a crucial role in this investigation as the energy transfer efficiency was found to change in presence of DPPC at different pH. It has been demonstrated that with proper calibration it is possible to use the present system under investigation to realize various ionic states of DPPC by observing the change in FRET efficiency between these two dyes. - Graphical abstract: Electrostatic interaction between anionic Flu and cationic R6G molecules in presence and absence of DPPC at different pH. Here pH of DPPC was changed, not the pH of individual dyes.

Development of a pan-Babesia FRET-qPCR and a survey of livestock from five Caribbean islands.

Science.gov (United States)

Li, Jing; Kelly, Patrick; Zhang, Jilei; Xu, Chuanling; Wang, Chengming

2015-09-30

Babesia spp. are tick-borne protozoan hemoparasites and the second most common blood-borne parasites of mammals, in particular domestic animals. We used the Clustal Multiple Alignment program and 18S rRNA gene sequences of 22 Babesia species from GenBank to develop a PCR that could detect a wide variety of Babesia spp. in a single reaction. The pan-Babesia FRET-qPCR we developed reliably detected B. gibsoni, B. canis, B. vogeli, B. microti, B. bovis, and B. divergens under controlled conditions but did not react with closely related species, mainly Hepatozoon americanum, Theileria equi, and Toxoplasma gondii. When we tested the pan-Babesia FRET-qPCR on DNA of whole blood from 752 cattle, sheep, goats, donkeys and horses from five Caribbean islands, we detected Babesia spp. expected to be present in the animals, mainly B. bovis and B. bigemina in cattle and B. caballi in horses and donkeys. Further, we found that animals were not uncommonly infected with species of Babesia usually associated with other hosts, mainly B. vogeli and B. gibsoni in cattle, sheep and goats, B. rossi in goats, and B. caballi in goats and sheep. Finally, the pan-Babesia FRET-qPCR enabled us to identify unknown species of Babesia in cattle, goats, sheep and donkeys. Overall, 70 % (525/752) of the animals we tested were positive confirming earlier limited studies that infections with Babesia spp. are common in livestock in the Caribbean.

Flow-induced vibration and fretting-wear specifications to ensure steam-generator and heat exchanger lifetime performance

International Nuclear Information System (INIS)

Janzen, V.P.; Han, Y.; Pettigrew, M.J.

2008-01-01

The current interest in refurbishment, life extension and new-build activity has meant a renewed emphasis on technical specifications that will ensure improved reliability and longer life. Preventing vibration and fretting-wear problems in steam generators and heat exchangers requires design specifications that bring together specific guidelines, analysis methods, requirements and appropriate performance criteria. The specifications must be firmly based on experimental data and field inspections. In addition, the specifications must be supported by theoretical analyses and fundamental scaling correlations, to cover conditions and geometries over the wide range applicable to existing components and probable future designs. The specifications are expected to evolve to meet changing industry requirements. This paper outlines the steps required to generate and support design specifications, and relates them to typical steam-generator design features and computer modeling capabilities. It also describes current issues that are driving changes to flow-induced vibration and fretting-wear specifications that can be applied to the design process for component refurbishment, replacement or new designs. These issues include recent experimental or field evidence for new excitation mechanisms, e.g., the possibility of in-plane fluidelastic instability of U-tubes, the demand for longer reactor and component lifetimes, the need for better predictions of dynamic properties and vibration response, e.g., two-phase random-turbulence excitation, and requirements to consider system 'excursions' or abnormal scenarios, e.g., a main steam line break in the case of steam generators. The paper describes steps being taken to resolve these issues. (author)

Characterization of a spectrally diverse set of fluorescent proteins as FRET acceptors for mTurquoise2

NARCIS (Netherlands)

Mastop, M.; Bindels, D.S.; Shaner, N.C.; Postma, M.; Gadella, T.W.J.; Goedhart, J.

2017-01-01

The performance of Förster Resonance Energy Transfer (FRET) biosensors depends on brightness and photostability, which are dependent on the characteristics of the fluorescent proteins that are employed. Yellow fluorescent protein (YFP) is often used as an acceptor but YFP is prone to photobleaching

Inter-Dye Distance Distributions Studied by a Combination of Single-Molecule FRET-Filtered Lifetime Measurements and a Weighted Accessible Volume (wAV Algorithm

Directory of Open Access Journals (Sweden)

Henning Höfig

2014-11-01

Full Text Available Förster resonance energy transfer (FRET is an important tool for studying the structural and dynamical properties of biomolecules. The fact that both the internal dynamics of the biomolecule and the movements of the biomolecule-attached dyes can occur on similar timescales of nanoseconds is an inherent problem in FRET studies. By performing single-molecule FRET-filtered lifetime measurements, we are able to characterize the amplitude of the motions of fluorescent probes attached to double-stranded DNA standards by means of flexible linkers. With respect to previously proposed experimental approaches, we improved the precision and the accuracy of the inter-dye distance distribution parameters by filtering out the donor-only population with pulsed interleaved excitation. A coarse-grained model is employed to reproduce the experimentally determined inter-dye distance distributions. This approach can easily be extended to intrinsically flexible proteins allowing, under certain conditions, to decouple the macromolecule amplitude of motions from the contribution of the dye linkers.

Quantitative standard-less XRF analysis

International Nuclear Information System (INIS)

Ulitzka, S.

2002-01-01

Full text: For most analytical tasks in the mining and associated industries matrix-matched calibrations are used for the monitoring of ore grades and process control. In general, such calibrations are product specific (iron ore, bauxite, alumina, mineral sands, cement etc.) and apply to a relatively narrow concentration range but give the best precision and accuracy for those materials. A wide range of CRMs is available and for less common materials synthetic standards can be made up from 'pure' chemicals. At times, analysis of materials with varying matrices (powders, scales, fly ash, alloys, polymers, liquors, etc.) and diverse physical shapes (non-flat, metal drillings, thin layers on substrates etc.) is required that could also contain elements which are not part of a specific calibration. A qualitative analysis can provide information about the presence of certain elements and the relative intensities of element peaks in a scan can give a rough idea about their concentrations. More often however, quantitative values are required. The paper will look into the basics of quantitative standardless analysis and show results for some well-defined CRMs. Copyright (2002) Australian X-ray Analytical Association Inc

Reactor applications of quantitative diffraction analysis

International Nuclear Information System (INIS)

Feguson, I.F.

1976-09-01

Current work in quantitative diffraction analysis was presented under the main headings of: thermal systems, fast reactor systems, SGHWR applications and irradiation damage. Preliminary results are included on a comparison of various new instrumental methods of boron analysis as well as preliminary new results on Zircaloy corrosion, and materials transfer in liquid sodium. (author)

Quantitative analysis of myocardial tissue with digital autofluorescence microscopy

Directory of Open Access Journals (Sweden)

Thomas Jensen

2016-01-01

Full Text Available Background: The opportunity offered by whole slide scanners of automated histological analysis implies an ever increasing importance of digital pathology. To go beyond the importance of conventional pathology, however, digital pathology may need a basic histological starting point similar to that of hematoxylin and eosin staining in conventional pathology. This study presents an automated fluorescence-based microscopy approach providing highly detailed morphological data from unstained microsections. This data may provide a basic histological starting point from which further digital analysis including staining may benefit. Methods: This study explores the inherent tissue fluorescence, also known as autofluorescence, as a mean to quantitate cardiac tissue components in histological microsections. Data acquisition using a commercially available whole slide scanner and an image-based quantitation algorithm are presented. Results: It is shown that the autofluorescence intensity of unstained microsections at two different wavelengths is a suitable starting point for automated digital analysis of myocytes, fibrous tissue, lipofuscin, and the extracellular compartment. The output of the method is absolute quantitation along with accurate outlines of above-mentioned components. The digital quantitations are verified by comparison to point grid quantitations performed on the microsections after Van Gieson staining. Conclusion: The presented method is amply described as a prestain multicomponent quantitation and outlining tool for histological sections of cardiac tissue. The main perspective is the opportunity for combination with digital analysis of stained microsections, for which the method may provide an accurate digital framework.

Sub-ensemble monitoring of DNA strand displacement using multiparameter single-molecule FRET

OpenAIRE

Baltierra Jasso, Laura; Morten, Michael; Magennis, Steven William

2018-01-01

Non-enzymatic DNA strand displacement is an important mechanism in dynamic DNA nanotechnology. Here we show that the large parameter space that is accessible by single-molecule FRET is ideal for the simultaneous monitoring of multiple reactants and products of DNA strand exchange reactions. We monitored the strand displacement from double-stranded DNA (dsDNA) by single-stranded DNA (ssDNA) at 37 °C; the data were modelled as a second-order reaction approaching equilibrium, with a rate constan...

Regulatory assembly of the vacuolar proton pump VoV1-ATPase in yeast cells by FLIM-FRET

Science.gov (United States)

Ernst, Stefan; Batisse, Claire; Zarrabi, Nawid; Böttcher, Bettina; Börsch, Michael

2010-02-01

We investigate the reversible disassembly of VOV1-ATPase in life yeast cells by time resolved confocal FRET imaging. VOV1-ATPase in the vacuolar membrane pumps protons from the cytosol into the vacuole. VOV1-ATPase is a rotary biological nanomotor driven by ATP hydrolysis. The emerging proton gradient is used for secondary transport processes as well as for pH and Ca2+ homoeostasis in the cell. The activity of the VOV1-ATPase is regulated through assembly / disassembly processes. During starvation the two parts of VOV1-ATPase start to disassemble. This process is reversed after addition of glucose. The exact mechanisms are unknown. To follow the disassembly / reassembly in vivo we tagged two subunits C and E with different fluorescent proteins. Cellular distributions of C and E were monitored using a duty cycle-optimized alternating laser excitation scheme (DCO-ALEX) for time resolved confocal FRET-FLIM measurements.

One-pot fabrication of FRET-based fluorescent probe for detecting copper ion and sulfide anion in 100% aqueous media

Science.gov (United States)

Lv, Kun; Chen, Jian; Wang, Hong; Zhang, Peisheng; Yu, Maolin; Long, Yunfei; Yi, Pinggui

2017-04-01

The design of effective tools for detecting copper ion (Cu2 +) and sulfide anion (S2 -) is of great importance due to the abnormal level of Cu2 + and S2 - has been associated with an increase in risk of many diseases. Herein, we report on the fabrication of fluorescence resonance energy transfer (FRET) based fluorescent probe PF (PEI-FITC) for detecting Cu2 + and S2 - in 100% aqueous media via a facile one-pot method by covalent linking fluorescein isothiocyanate (FITC) with branched-polyethylenimine (b-PEI). PF could selectively coordinate with Cu2 + among 10 metal ions to form PF-Cu2 + complex, resulting in fluorescence quenching through FRET mechanism. Furthermore, the in situ generated PF-Cu2 + complex can be used to selectively detect S2 - based on the displacement approach, resulting in an off-on type sensing. There is no obvious interference from other anions, such as Cl-, NO3-, ClO4-, SO42 -, HCO3-, CO32 -, Br-, HPO42 -, F- and S2O32 -. In addition, PF was successfully used to determine Cu2 + and S2 - in human serum and tap water samples. Therefore, the FRET-based probe PF may provide a new method for selective detection of multifarious analysts in biological and environmental applications, and even hold promise for application in more complicated systems.

Quantitative risk analysis of a space shuttle subsystem

International Nuclear Information System (INIS)

Frank, M.V.

1989-01-01

This paper reports that in an attempt to investigate methods for risk management other than qualitative analysis techniques, NASA has funded pilot study quantitative risk analyses for space shuttle subsystems. The authors performed one such study of two shuttle subsystems with McDonnell Douglas Astronautics Company. The subsystems were the auxiliary power units (APU) on the orbiter, and the hydraulic power units on the solid rocket booster. The technology and results of the APU study are presented in this paper. Drawing from a rich in-flight database as well as from a wealth of tests and analyses, the study quantitatively assessed the risk of APU-initiated scenarios on the shuttle during all phases of a flight mission. Damage states of interest were loss of crew/vehicle, aborted mission, and launch scrub. A quantitative risk analysis approach to deciding on important items for risk management was contrasted with the current NASA failure mode and effects analysis/critical item list approach

Uncertainty of quantitative microbiological methods of pharmaceutical analysis.

Science.gov (United States)

Gunar, O V; Sakhno, N G

2015-12-30

The total uncertainty of quantitative microbiological methods, used in pharmaceutical analysis, consists of several components. The analysis of the most important sources of the quantitative microbiological methods variability demonstrated no effect of culture media and plate-count techniques in the estimation of microbial count while the highly significant effect of other factors (type of microorganism, pharmaceutical product and individual reading and interpreting errors) was established. The most appropriate method of statistical analysis of such data was ANOVA which enabled not only the effect of individual factors to be estimated but also their interactions. Considering all the elements of uncertainty and combining them mathematically the combined relative uncertainty of the test results was estimated both for method of quantitative examination of non-sterile pharmaceuticals and microbial count technique without any product. These data did not exceed 35%, appropriated for a traditional plate count methods. Copyright © 2015 Elsevier B.V. All rights reserved.

A rhodamine–dansyl conjugate as a FRET based sensor for Fe{sup 3+} in the red spectral region

Energy Technology Data Exchange (ETDEWEB)

Xie, Puhui, E-mail: pxie2007@yahoo.com.cn [College of Sciences, Henan Agricultural University, Zhengzhou 450002 (China); Guo, Fengqi, E-mail: fqguo@zzu.edu.cn [College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Xia, Ruirui; Wang, Yao [College of Sciences, Henan Agricultural University, Zhengzhou 450002 (China); Yao, Denghui [College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Yang, Guoyu; Xie, Lixia [College of Sciences, Henan Agricultural University, Zhengzhou 450002 (China)

2014-01-15

A new fluorescent resonance energy transfer (FRET) based fluorescent probe (compound 1) containing a dansyl unit as a donor and rhodamine 101 as an acceptor was developed to detect Fe{sup 3+} from other transition metal ions through ratiometric sensing in organic-aqueous solutions. Fe{sup 3+} induced a ring-opening reaction of the spirolactam rhodamine moiety of 1 resulting in the formation of a fluorescent derivative that can serve as the FRET acceptor. Ratiometric sensing of Fe{sup 3+} was accomplished by plotting the fluorescence intensity ratio at 605 nm and 515 nm versus ferric ion concentration. The probe displayed a linear response to Fe{sup 3+} in the range of 5.5–25 μM with a detection limit of 0.64 μM. A 1:1 stoichiometry for the 1–Fe{sup 3+} complex was formed with an association constant of 1.74×10{sup 4} M{sup −1}. The probe also exhibited a large Stokes shift (225 nm) which can eliminate backscattering effects of excitation light. -- Highlights: • A new colorimetric and fluorescent “off–on” chemosensor for Fe{sup 3+} was synthesized. • It can respond to Fe{sup 3+} in the red spectral region based on a FRET mechanism. • Its ratiometric sensing for Fe{sup 3+} can be accomplished with a signal to noise ratio of 214. • The large Stokes shift (225 nm) can rule out the excitation backscattering effects.

Förster Resonance Energy Transfer (FRET) between Heterogeneously Distributed Probes: Application to Lipid Nanodomains and Pores

Czech Academy of Sciences Publication Activity Database

Šachl, Radek; Johansson, L. B. A.; Hof, Martin

2012-01-01

Roč. 13, č. 12 (2012), s. 16141-16156 E-ISSN 1422-0067 R&D Projects: GA ČR GAP208/10/1090; GA ČR GAP208/10/0376 Institutional support: RVO:61388955 Keywords : FRET * lipid domains * pores Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.464, year: 2012

Theoretical-experimental assessment of the variables affecting fretting of Atucha I nuclear power plant utility steam generators tubes

International Nuclear Information System (INIS)

Kulichevsky, Raul M.

1995-01-01

Fretting wear of Steam Generator tubes caused by flow induced vibrations generates uncertainty on their integrity. The knowledge of the controlling variables of the wear process may give a criterion to evaluate the tubes residual life. Information on vibratory response and dynamic interaction between tubes and their supports are prerequisites for understanding the relationship between fretting wear and tube vibration. Experimental results of the vibratory response of an Atucha-I nuclear power plant type U-tube, the influence of tube/support clearance on this response and a study of tube/support dynamic interaction, which allow the verification of a finite element model of this type of tubes, are presented in this work. Also wear results for the Incoloy 800/DIN 1.4550 austenitic stainless steel pair of materials and a first evaluation of the wear constant of this pair are presented. (author)

Quantitative Analysis of cardiac SPECT

International Nuclear Information System (INIS)

Nekolla, S.G.; Bengel, F.M.

2004-01-01

The quantitative analysis of myocardial SPECT images is a powerful tool to extract the highly specific radio tracer uptake in these studies. If compared to normal data bases, the uptake values can be calibrated on an individual basis. Doing so increases the reproducibility of the analysis substantially. Based on the development over the last three decades starting from planar scinitigraphy, this paper discusses the methods used today incorporating the changes due to tomographic image acquisitions. Finally, the limitations of these approaches as well as consequences from most recent hardware developments, commercial analysis packages and a wider view of the description of the left ventricle are discussed. (orig.)

Good practices for quantitative bias analysis.

Science.gov (United States)

Lash, Timothy L; Fox, Matthew P; MacLehose, Richard F; Maldonado, George; McCandless, Lawrence C; Greenland, Sander

2014-12-01

Quantitative bias analysis serves several objectives in epidemiological research. First, it provides a quantitative estimate of the direction, magnitude and uncertainty arising from systematic errors. Second, the acts of identifying sources of systematic error, writing down models to quantify them, assigning values to the bias parameters and interpreting the results combat the human tendency towards overconfidence in research results, syntheses and critiques and the inferences that rest upon them. Finally, by suggesting aspects that dominate uncertainty in a particular research result or topic area, bias analysis can guide efficient allocation of sparse research resources. The fundamental methods of bias analyses have been known for decades, and there have been calls for more widespread use for nearly as long. There was a time when some believed that bias analyses were rarely undertaken because the methods were not widely known and because automated computing tools were not readily available to implement the methods. These shortcomings have been largely resolved. We must, therefore, contemplate other barriers to implementation. One possibility is that practitioners avoid the analyses because they lack confidence in the practice of bias analysis. The purpose of this paper is therefore to describe what we view as good practices for applying quantitative bias analysis to epidemiological data, directed towards those familiar with the methods. We focus on answering questions often posed to those of us who advocate incorporation of bias analysis methods into teaching and research. These include the following. When is bias analysis practical and productive? How does one select the biases that ought to be addressed? How does one select a method to model biases? How does one assign values to the parameters of a bias model? How does one present and interpret a bias analysis?. We hope that our guide to good practices for conducting and presenting bias analyses will encourage

Quantitative phase analysis in industrial research

International Nuclear Information System (INIS)

Ahmad Monshi

1996-01-01

X-Ray Diffraction (XRD) is the only technique able to identify phase and all the other analytical techniques give information about the elements. Quantitative phase analysis of minerals and industrial products is logically the next step after a qualitative examination and is of great importance in industrial research. Since the application of XRD in industry, early in this century, workers were trying to develop quantitative XRD methods. In this paper some of the important methods are briefly discussed and partly compared. These methods are Internal Standard, Known Additions, Double Dilution, External Standard, Direct Comparison, Diffraction Absorption and Ratio of Slopes

Compound parabolic concentrator optical fiber tip for FRET-based fluorescent sensors

DEFF Research Database (Denmark)

Hassan, Hafeez Ul; Nielsen, Kristian; Aasmul, Soren

2015-01-01

The Compound Parabolic Concentrator (CPC) optical fiber tip shape has been proposed for intensity based fluorescent sensors working on the principle of FRET (Förster Resonance Energy Transfer). A simple numerical Zemax model has been used to optimize the CPC tip geometry for a step-index multimode...... polymer optical fiber for an excitation and emission wavelength of 550 nm and 650nm, respectively. The model suggests an increase of a factor of 1.6 to 4 in the collected fluorescent power for an ideal CPC tip, as compared to the plane-cut fiber tip for fiber lengths between 5 and 45mm...

TECHNIQUE OF TESTING ON FRETTING AT THE SPHERE-TO-PLANE CONTACT

Directory of Open Access Journals (Sweden)

А. Khimko

2012-12-01

Full Text Available  The methodology of conducting tests on fretting at the sphere-to-plane contact was developed for the wing mechanization unit, namely for screw-nut pair with intermediate balls. Wearability tests were conducted on a modified installation МФК-1, the feature of which is the designed holder that allows testing with real balls. It was found that at the dry contact of ШХ-15 and 30Х2НВФA materials, surface microcracks are formed due to welding of microasperities areas and their rupture under the influence of vibration.

Probing nucleic acid interactions and pre-mRNA splicing by Förster resonance energy transfer (FRET) microscopy

Czech Academy of Sciences Publication Activity Database

Šimková, Eva; Staněk, David

2012-01-01

Roč. 13, č. 11 (2012), s. 14929-14945 E-ISSN 1422-0067 R&D Projects: GA AV ČR KAN200520801 Institutional support: RVO:68378050 Keywords : FRET * FLIM * acceptor photobleaching * RNA interactions * spliceosome Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 2.464, year: 2012

FRET-based binding assay between a fluorescent cAMP analogue and a cyclic nucleotide-binding domain tagged with a CFP.

Science.gov (United States)

Romero, Francisco; Santana-Calvo, Carmen; Sánchez-Guevara, Yoloxochitl; Nishigaki, Takuya

2017-09-01

The cyclic nucleotide-binding domain (CNBD) functions as a regulatory domain of many proteins involved in cyclic nucleotide signalling. We developed a straightforward and reliable binding assay based on intermolecular fluorescence resonance energy transfer (FRET) between an adenosine-3', 5'-cyclic monophosphate analogue labelled with fluorescein and a recombinant CNBD of human EPAC1 tagged with a cyan fluorescence protein (CFP). The high FRET efficiency of this method (~ 80%) allowed us to perform several types of binding experiments with nanomolar range of sample using conventional equipment. In addition, the CFP tag on the CNBD enabled us to perform a specific binding experiment using an unpurified protein. Considering these advantages, this technique is useful to study poorly characterized CNBDs. © 2017 Federation of European Biochemical Societies.

Effects of the different frequencies and loads of ultrasonic surface rolling on surface mechanical properties and fretting wear resistance of HIP Ti–6Al–4V alloy

Energy Technology Data Exchange (ETDEWEB)

Li, G., E-mail: ligang_scut@outlook.com; Qu, S.G., E-mail: qusg@scut.edu.cn; Pan, Y.X.; Li, X.Q.

2016-12-15

Highlights: • Effects of MUSR frequency and load on surface properties of HIP Ti-6Al-4V investigated. • The grains in surface-modified layer were refined and appeared twins and many dense dislocations. • The hardened layer depth and surface residual stress of MUSR- treated samples were significantly improved. • MUSR- treated samples showed the good fretting friction and wear resistance. • The best microstructure and properties of surface-modified layer obtained by sample treated by 30 kHz and 900 N. - Abstract: The main purpose of this paper was to investigate the effects of the different frequencies and loads of multi-pass ultrasonic surface rolling (MUSR) on surface layer mechanical properties, microstructure and fretting friction and wear characteristics of HIP (hot isostatic pressing) Ti–6Al–4 V alloy. Some microscopic analysis methods (SEM, TEM and EDS) were used to characterize the modified surface layer of material after MUSR treatment. The results indicated that the material in sample surface layer experienced a certain extent plastic deformation, and accompanied by some dense dislocations and twins generation. Moreover surface microhardness and residual stress of samples treated by MUSR were also greatly improved compared with the untreated. The fretting friction and wear properties of samples treated by MUSR in different conditions are tested at 10 and 15 N in dry friction conditions. It could be found that friction coefficient and wear volume loss were significantly declined in the optimal result. The main wear mechanism of MUSR-treated samples included abrasive wear, adhesion and spalling.

Micro-computer system for quantitative image analysis of damage microstructure

International Nuclear Information System (INIS)

Kohyama, A.; Kohno, Y.; Satoh, K.; Igata, N.

1984-01-01

Quantitative image analysis of radiation induced damage microstructure is very important in evaluating material behaviors in radiation environment. But, quite a few improvement have been seen in quantitative analysis of damage microstructure in these decades. The objective of this work is to develop new system for quantitative image analysis of damage microstructure which could improve accuracy and efficiency of data sampling and processing and could enable to get new information about mutual relations among dislocations, precipitates, cavities, grain boundaries, etc. In this system, data sampling is done with X-Y digitizer. The cavity microstructure in dual-ion irradiated 316 SS is analyzed and the effectiveness of this system is discussed. (orig.)

The role of two-phase coolant in moderating fretting in nuclear steam generators

International Nuclear Information System (INIS)

Dyke, J.M.

2004-01-01

This paper expands the principal of coolant-cushioning in Nuclear Steam Generators whereby the two-phase coolant, especially the bubble film on the tube surface, moderates the vibration of coolant tubes against their supports. The current paper addresses tube bundle and anti-vibration bars (AVB) geometry issues; examines the tube bundle-coolant-AVB interfaces and examines implications for recirculation flow, AVB design and boiler size. In a T(sat) fluid, the tube surface is uniformly coating with growing bubbles whose momentum is perpendicular to the surface at first, then they are swept away by the bulk flow. The combination of this momentum, the phase change and the water film remaining on the surface, counteract the vibration energy of the tube-AVB system, reducing the likelihood of metal-to-metal contact and consequent fretting. To maximize the benefit of the cushioning effect, the following design inputs are needed: 1) the AVB-tube interface should have sufficient clearance for the T(sat) solution to operate, 2) The AVB should be wide enough to generate the necessary cushioning force, and 3) the AVB should be thin enough to be flexible and absorb some of the transferred vibration energy. Furthermore, fretting and crude deposition at the AVB-tube interface can be reduced or eliminated by reducing the number of AVBs, increasing clearances and making the AVBs limber

Quantitative Structure-Activity Relationship Analysis of the ...

African Journals Online (AJOL)

Erah

Quantitative Structure-Activity Relationship Analysis of the Anticonvulsant ... Two types of molecular descriptors, including the 2D autocorrelation ..... It is based on the simulation of natural .... clustering anticonvulsant, antidepressant, and.

Failure of fretted steam generator tubes under accident conditions

International Nuclear Information System (INIS)

Forrest, C.F.

1996-10-01

Tests were carried out with a bank of tubes in a water tunnel to determine the tolerance of flawed nuclear reactor steam generator tubes to accident conditions which would result in high cross-flow velocities. Fourteen specimen tubes were tested, each having one or two types of defect machined into the surface simulating fretting-wear type scars found in some operating steam generators. The tubes were tested at flow velocities sufficient to induce high fluid elastic-type vibrations. Seven of the tubes failed near the thinnest section of the defects during the one-hour tests, due to impacting and/or rubbing between the tube and the support. Strain gauges, displacement transducers, force gauges and an accelerometer were used on the target tube and/or the tube immediately downstream of it to measure their vibrational characteristics

Compact quantum dot-antibody conjugates for FRET immunoassays with subnanomolar detection limits

Science.gov (United States)

Mattera, Lucia; Bhuckory, Shashi; Wegner, K. David; Qiu, Xue; Agnese, Fabio; Lincheneau, Christophe; Senden, Tim; Djurado, David; Charbonnière, Loïc J.; Hildebrandt, Niko; Reiss, Peter

2016-05-01

A novel two-step approach for quantum dot (QD) functionalization and bioconjugation is presented, which yields ultra-compact, stable, and highly luminescent antibody-QD conjugates suitable for use in FRET immunoassays. Hydrophobic InPZnS/ZnSe/ZnS (emission wavelength: 530 nm), CdSe/ZnS (605 nm), and CdSeTe/ZnS (705 nm) QDs were surface functionalized with zwitterionic penicillamine, enabling aqueous phase transfer under conservation of the photoluminescence properties. Post-functionalization with a heterobifunctional crosslinker, containing a lipoic acid group and a maleimide function, enabled the subsequent coupling to sulfhydryl groups of proteins. This was demonstrated by QD conjugation with fragmented antibodies (F(ab)). The obtained F(ab)-QD conjugates range among the smallest antibody-functionalized nanoprobes ever reported, with a hydrodynamic diameter coating for FRET could be demonstrated by an 6.2 and 2.5 fold improvement of the limit of detection (LOD) for PSA compared to commercially available hydrophilic QDs emitting at 605 and 705 nm, respectively. While the commercial QDs contain identical inorganic cores responsible for their fluorescence, they are coated with a comparably thick amphiphilic polymer layer leading to much larger hydrodynamic diameters (>26 nm without biomolecules). The LODs of 0.8 and 3.7 ng mL-1 obtained in 50 μL serum samples are below the clinical cut-off level of PSA (4 ng mL-1) and demonstrate their direct applicability in clinical diagnostics.A novel two-step approach for quantum dot (QD) functionalization and bioconjugation is presented, which yields ultra-compact, stable, and highly luminescent antibody-QD conjugates suitable for use in FRET immunoassays. Hydrophobic InPZnS/ZnSe/ZnS (emission wavelength: 530 nm), CdSe/ZnS (605 nm), and CdSeTe/ZnS (705 nm) QDs were surface functionalized with zwitterionic penicillamine, enabling aqueous phase transfer under conservation of the photoluminescence properties. Post

Data from quantitative label free proteomics analysis of rat spleen

Directory of Open Access Journals (Sweden)

Khadar Dudekula

2016-09-01

Full Text Available The dataset presented in this work has been obtained using a label-free quantitative proteomic analysis of rat spleen. A robust method for extraction of proteins from rat spleen tissue and LC-MS-MS analysis was developed using a urea and SDS-based buffer. Different fractionation methods were compared. A total of 3484 different proteins were identified from the pool of all experiments run in this study (a total of 2460 proteins with at least two peptides. A total of 1822 proteins were identified from nine non-fractionated pulse gels, 2288 proteins and 2864 proteins were identified by SDS-PAGE fractionation into three and five fractions respectively. The proteomics data are deposited in ProteomeXchange Consortium via PRIDE PXD003520, Progenesis and Maxquant output are presented in the supported information. The generated list of proteins under different regimes of fractionation allow assessing the nature of the identified proteins; variability in the quantitative analysis associated with the different sampling strategy and allow defining a proper number of replicates for future quantitative analysis. Keywords: Spleen, Rat, Protein extraction, Label-free quantitative proteomics

Quantitative data analysis in education a critical introduction using SPSS

CERN Document Server

Connolly, Paul

2007-01-01

This book provides a refreshing and user-friendly guide to quantitative data analysis in education for students and researchers. It assumes absolutely no prior knowledge of quantitative methods or statistics. Beginning with the very basics, it provides the reader with the knowledge and skills necessary to be able to undertake routine quantitative data analysis to a level expected of published research. Rather than focusing on teaching statistics through mathematical formulae, the book places an emphasis on using SPSS to gain a real feel for the data and an intuitive grasp of t

Quantitative analysis method for ship construction quality

Directory of Open Access Journals (Sweden)

FU Senzong

2017-03-01

Full Text Available The excellent performance of a ship is assured by the accurate evaluation of its construction quality. For a long time, research into the construction quality of ships has mainly focused on qualitative analysis due to a shortage of process data, which results from limited samples, varied process types and non-standardized processes. Aiming at predicting and controlling the influence of the construction process on the construction quality of ships, this article proposes a reliability quantitative analysis flow path for the ship construction process and fuzzy calculation method. Based on the process-quality factor model proposed by the Function-Oriented Quality Control (FOQC method, we combine fuzzy mathematics with the expert grading method to deduce formulations calculating the fuzzy process reliability of the ordinal connection model, series connection model and mixed connection model. The quantitative analysis method is applied in analyzing the process reliability of a ship's shaft gear box installation, which proves the applicability and effectiveness of the method. The analysis results can be a useful reference for setting key quality inspection points and optimizing key processes.

Quantitative X-ray analysis of pigments

International Nuclear Information System (INIS)

Araujo, M. Marrocos de

1987-01-01

The 'matrix-flushing' and the 'adiabatic principle' methods have been applied for the quantitative analysis through X-ray diffraction patterns of pigments and extenders mixtures, frequently used in paint industry. The results obtained have shown the usefulness of these methods, but still ask for improving their accuracy. (Author) [pt

An automated wide-field time-gated optically sectioning fluorescence lifetime imaging multiwell plate reader for high-content analysis of protein-protein interactions

Science.gov (United States)

Alibhai, Dominic; Kumar, Sunil; Kelly, Douglas; Warren, Sean; Alexandrov, Yuriy; Munro, Ian; McGinty, James; Talbot, Clifford; Murray, Edward J.; Stuhmeier, Frank; Neil, Mark A. A.; Dunsby, Chris; French, Paul M. W.

2011-03-01

We describe an optically-sectioned FLIM multiwell plate reader that combines Nipkow microscopy with wide-field time-gated FLIM, and its application to high content analysis of FRET. The system acquires sectioned FLIM images in fluorescent protein. It has been applied to study the formation of immature HIV virus like particles (VLPs) in live cells by monitoring Gag-Gag protein interactions using FLIM FRET of HIV-1 Gag transfected with CFP or YFP. VLP formation results in FRET between closely packed Gag proteins, as confirmed by our FLIM analysis that includes automatic image segmentation.

Application of contact mechanics for fretting damage of fuel rod: part 1 influence functions and numerical method

International Nuclear Information System (INIS)

Kim, H. K.; Yoon, K. H.; Kang, H. S.; Song, G. N.

1998-01-01

For the analysis of the fretting problem of the fuel rods, present paper(Part I) shows the numerical method developed for evaluating the stresses on the contact surfaces between the fuel rods and the spacer grids. Theory of Contact Mechanics was incorporated. Contact area was regarded as a plane strain condition, so plane problem was taken into consideration. Normal stress profile on the contact surface was assumed to be Hertzian. As for the direction of the shear load, a closed load path, e.g. load increase in transverse increase in axial decrease in transverse decrease in axial increase in transverse increase in axial direction was considered for simulating the rod vibration in a reactor core. Partial slip problem was consulted. As for the numerical method, a triangular traction element was utilized and the corresponding influence functions were evaluated. Numerical program has been implemented for the present analysis, of which the validity was verified by comparing the Mindlin-Cattaneo solution

Novel approach in quantitative analysis of shearography method

International Nuclear Information System (INIS)

Wan Saffiey Wan Abdullah

2002-01-01

The application of laser interferometry in industrial non-destructive testing and material characterization is becoming more prevalent since this method provides non-contact full-field inspection of the test object. However their application only limited to the qualitative analysis, current trend has changed to the development of this method by the introduction of quantitative analysis, which attempts to detail the defect examined. This being the design feature for a ranges of object size to be examined. The growing commercial demand for quantitative analysis for NDT and material characterization is determining the quality of optical and analysis instrument. However very little attention is currently being paid to understanding, quantifying and compensating for the numerous error sources which are a function of interferometers. This paper presents a comparison of measurement analysis using the established theoretical approach and the new approach, taken into account the factor of divergence illumination and other geometrical factors. The difference in the measurement system could be associated in the error factor. (Author)

Expectation-maximization of the potential of mean force and diffusion coefficient in Langevin dynamics from single molecule FRET data photon by photon.

Science.gov (United States)

Haas, Kevin R; Yang, Haw; Chu, Jhih-Wei

2013-12-12

The dynamics of a protein along a well-defined coordinate can be formally projected onto the form of an overdamped Lagevin equation. Here, we present a comprehensive statistical-learning framework for simultaneously quantifying the deterministic force (the potential of mean force, PMF) and the stochastic force (characterized by the diffusion coefficient, D) from single-molecule Förster-type resonance energy transfer (smFRET) experiments. The likelihood functional of the Langevin parameters, PMF and D, is expressed by a path integral of the latent smFRET distance that follows Langevin dynamics and realized by the donor and the acceptor photon emissions. The solution is made possible by an eigen decomposition of the time-symmetrized form of the corresponding Fokker-Planck equation coupled with photon statistics. To extract the Langevin parameters from photon arrival time data, we advance the expectation-maximization algorithm in statistical learning, originally developed for and mostly used in discrete-state systems, to a general form in the continuous space that allows for a variational calculus on the continuous PMF function. We also introduce the regularization of the solution space in this Bayesian inference based on a maximum trajectory-entropy principle. We use a highly nontrivial example with realistically simulated smFRET data to illustrate the application of this new method.

Effects of Contact Load on the Fretting Fatigue Behavior of IN-100 at Elevated Temperature

Science.gov (United States)

2009-03-01

Effect of contact pressure on fretting fatigue of austenitic stainless steel ,” Tribology International, vol. 36, pp. 79-85, 2003. 155 [56] N.K. Naidu...austenitic stainless steel was presented. Like the studies in the previous section, this study investigated how a variably increased contact load...that their stainless steel specimens acted much in the same manner as the aluminum specimens presented in the previous section. It was observed

Sliding Wear and Fretting Wear of DLC-Based, Functionally Graded Nanocomposite Coatings

Science.gov (United States)

Miyoshi, K.; Pohlchuck, B.; Street, Kenneth W.; Zabinski, J. S.; Sanders, J. H.; Voevodin, A. a.; Wu, R. L. C.

1999-01-01

Improving the tribological functionality of diamondlike carbon (DLC) films--developing, good wear resistance, low friction, and high load-carrying capacity-was the aim of this investigation. Nanocomposite coatings consisting of an amorphous DLC (a-DLC) top layer and a functionally graded titanium-titanium carbon-diamondlike carbon (Ti-Ti(sub x) C(sub y)-DLC) underlayer were produced on AISI 440C stainless steel substrates by the hybrid technique of magnetron sputtering and pulsed-laser deposition. The resultant DLC films were characterized by Raman spectroscopy, scanning electron microscopy, and surface profilometry. Two types of wear experiment were conducted in this investioation: sliding friction experiments and fretting wear experiments. Unidirectional ball-on-disk sliding friction experiments were conducted to examine the wear behavior of an a-DLC/Ti-Ti(sub x) C(sub y)-DLC-coated AISI 440C stainless steel disk in sliding contact with a 6-mm-diameter AISI 440C stainless steel ball in ultrahigh vacuum, dry nitrogen, and humid air. Although the wear rates for both the coating and ball were low in all three environments, the humid air and dry nitrogen caused mild wear with burnishing, in the a-DLC top layer, and the ultrahigh vacuum caused relatively severe wear with brittle fracture in both the a-DLC top layer and the Ti-Ti(sub x) C(sub y)-DLC underlayer. For reference, amorphous hydrogenated carbon (H-DLC) films produced on a-DLC/Ti-Ti(sub x) C(sub y)-DLC nanocomposite coatings by using an ion beam were also examined in the same manner. The H-DLC films markedly reduced friction even in ultrahigh vacuum without sacrificing wear resistance. The H-DLC films behaved much like the a-DLC/Ti-Ti(sub x) C(sub y)-DLC nanocomposite coating in dry nitrogen and humid air, presenting low friction and low wear. Fretting wear experiments were conducted in humid air (approximately 50% relative humidity) at a frequency of 80 Hz and an amplitude of 75 micron on an a

Quantitative methods for the analysis of electron microscope images

DEFF Research Database (Denmark)

Skands, Peter Ulrik Vallø

1996-01-01

The topic of this thesis is an general introduction to quantitative methods for the analysis of digital microscope images. The images presented are primarily been acquired from Scanning Electron Microscopes (SEM) and interfermeter microscopes (IFM). The topic is approached though several examples...... foundation of the thesis fall in the areas of: 1) Mathematical Morphology; 2) Distance transforms and applications; and 3) Fractal geometry. Image analysis opens in general the possibility of a quantitative and statistical well founded measurement of digital microscope images. Herein lies also the conditions...

Parameter determination for quantitative PIXE analysis using genetic algorithms

International Nuclear Information System (INIS)

Aspiazu, J.; Belmont-Moreno, E.

1996-01-01

For biological and environmental samples, PIXE technique is in particular advantage for elemental analysis, but the quantitative analysis implies accomplishing complex calculations that require the knowledge of more than a dozen parameters. Using a genetic algorithm, the authors give here an account of the procedure to obtain the best values for the parameters necessary to fit the efficiency for a X-ray detector. The values for some variables involved in quantitative PIXE analysis, were manipulated in a similar way as the genetic information is treated in a biological process. The authors carried out the algorithm until they reproduce, within the confidence interval, the elemental concentrations corresponding to a reference material

Fretting Fatigue Behaviour of Pin-Loaded Thermoset Carbon-Fibre-Reinforced Polymer (CFRP Straps

Directory of Open Access Journals (Sweden)

Fabio Baschnagel

2016-04-01

Full Text Available This paper focuses on the fretting fatigue behaviour of pin-loaded carbon-fibre-reinforced polymer (CFRP straps studied as models for rigging systems in sailing yachts, for suspenders of arch bridges and for pendent cables in cranes. Eight straps were subjected to an ultimate tensile strength test. In total, 26 straps were subjected to a fretting fatigue test, of which ten did not fail. An S–N curve was generated for a load ratio R of 0.1 and a frequency f of 10 Hz, showing a fatigue limit stress of the straps around the matrix fatigue limit, corresponding to 46% of the straps’ ultimate tensile strength (σUTS. The fatigue limit was defined as 3 million load cycles (N = 3 × 106, but tests were even conducted up to N = 11.09 × 106. Catastrophic failure of the straps was initiated in their vertex areas. Investigations on the residual strength and stiffness properties of straps tested around the fatigue limit stress (for N ≥ 1 × 106 showed little influence of the fatigue loading on these properties. Quasi-static finite element analyses (FEA were conducted. The results obtained from the FEA are in good agreement with the experiments and demonstrate a fibre parallel stress concentration in the vertex area of factor 1.3, under the realistic assumption of a coefficient of friction (cof between pin and strap of 0.5.

The 1.6 Å resolution structure of a FRET-optimized Cerulean fluorescent protein

Energy Technology Data Exchange (ETDEWEB)

Watkins, Jennifer L.; Kim, Hanseong [Arizona State University, Tempe, AZ 85287-1604 (United States); Markwardt, Michele L. [University of Maryland School of Medicine, Baltimore, MD 21201-1559 (United States); Chen, Liqing; Fromme, Raimund [Arizona State University, Tempe, AZ 85287-1604 (United States); Rizzo, Mark A. [University of Maryland School of Medicine, Baltimore, MD 21201-1559 (United States); Wachter, Rebekka M., E-mail: rwachter@asu.edu [Arizona State University, Tempe, AZ 85287-1604 (United States)

2013-05-01

The high resolution X-ray structure of the cyan fluorescent protein mCerulean3 demonstrates that different combinations of correlated residue substitutions can provide near optimum quantum yield values for fluorescence. Genetically encoded cyan fluorescent proteins (CFPs) bearing a tryptophan-derived chromophore are commonly used as energy-donor probes in Förster resonance energy transfer (FRET) experiments useful in live cell-imaging applications. In recent years, significant effort has been expended on eliminating the structural and excited-state heterogeneity of these proteins, which has been linked to undesirable photophysical properties. Recently, mCerulean3, a descendant of enhanced CFP, was introduced as an optimized FRET donor protein with a superior quantum yield of 0.87. Here, the 1.6 Å resolution X-ray structure of mCerulean3 is reported. The chromophore is shown to adopt a planar trans configuration at low pH values, indicating that the acid-induced isomerization of Cerulean has been eliminated. β-Strand 7 appears to be well ordered in a single conformation, indicating a loss of conformational heterogeneity in the vicinity of the chromophore. Although the side chains of Ile146 and Leu167 appear to exist in two rotamer states, they are found to be well packed against the indole group of the chromophore. The Ser65 reversion mutation allows improved side-chain packing of Leu220. A structural comparison with mTurquoise2 is presented and additional engineering strategies are discussed.

The 1.6 Å resolution structure of a FRET-optimized Cerulean fluorescent protein

International Nuclear Information System (INIS)

Watkins, Jennifer L.; Kim, Hanseong; Markwardt, Michele L.; Chen, Liqing; Fromme, Raimund; Rizzo, Mark A.; Wachter, Rebekka M.

2013-01-01

The high resolution X-ray structure of the cyan fluorescent protein mCerulean3 demonstrates that different combinations of correlated residue substitutions can provide near optimum quantum yield values for fluorescence. Genetically encoded cyan fluorescent proteins (CFPs) bearing a tryptophan-derived chromophore are commonly used as energy-donor probes in Förster resonance energy transfer (FRET) experiments useful in live cell-imaging applications. In recent years, significant effort has been expended on eliminating the structural and excited-state heterogeneity of these proteins, which has been linked to undesirable photophysical properties. Recently, mCerulean3, a descendant of enhanced CFP, was introduced as an optimized FRET donor protein with a superior quantum yield of 0.87. Here, the 1.6 Å resolution X-ray structure of mCerulean3 is reported. The chromophore is shown to adopt a planar trans configuration at low pH values, indicating that the acid-induced isomerization of Cerulean has been eliminated. β-Strand 7 appears to be well ordered in a single conformation, indicating a loss of conformational heterogeneity in the vicinity of the chromophore. Although the side chains of Ile146 and Leu167 appear to exist in two rotamer states, they are found to be well packed against the indole group of the chromophore. The Ser65 reversion mutation allows improved side-chain packing of Leu220. A structural comparison with mTurquoise2 is presented and additional engineering strategies are discussed

Optical bar code recognition of methyl salicylate (MES) for environmental monitoring using fluorescence resonance energy transfer (FRET) on thin films

Science.gov (United States)

Smith, Clint; Tatineni, Balaji; Anderson, John; Tepper, Gary

2006-10-01

Fluorescence resonance energy transfer (FRET) is a process in which energy is transferred nonradiatively from one fluorophore (the donor) in an excited electron state to another, the chromophore (the acceptor). FRET is distinctive in its ability to reveal the presence of specific recognition of select targets such as the nerve agent stimulant Methyl Salicylate (MES) upon spectroscopic excitation. We introduce a surface imprinted and non-imprinted thin film that underwent AC-Electrospray ionization for donor-acceptor pair(s) bound to InGaP quantum dots and mesoporous silicate nanoparticles. The donor-acceptor pair used in this investigation included MES (donor) and 6-(fluorescein-5-(and-6)- carboxamido) hexanoic acid, succinimidyl ester bound to InGaP quantum dots (acceptor). MES was then investigated as a donor to various acceptor fluorophore: InGaP: mesoporous silicate nanoparticle layers.

Towards characterization of DNA structure under physiological conditions in vivo at the single-molecule level using single-pair FRET

Czech Academy of Sciences Publication Activity Database

Fessl, Tomáš; Adamec, František; Polívka, Tomáš; Foldynová-Trantírková, Silvie; Vácha, František; Trantírek, L.

2012-01-01

Roč. 40, č. 16 (2012), s. 10 ISSN 0305-1048 Institutional research plan: CEZ:AV0Z50510513; CEZ:AV0Z60220518 Keywords : in-cell FRET * fluorescence * DNA * nucleic acid * ATTO * in vivo Subject RIV: BO - Biophysics Impact factor: 8.278, year: 2012

Modeling of fuel bundle vibration and the associated fretting wear in a CANDU fuel channel

International Nuclear Information System (INIS)

Mohany, A.; Hassan, M.

2011-01-01

In this paper a numerical model is developed to predict the vibration response of a CANDU® fuel bundle and the associated fretting wear in the surrounding pressure tube. One excitation mechanism is considered in this model; turbulence-induced excitation caused by coolant flow inside the fuel channel. The numerical model can be easily adapted to include the effects of seismic events, fuel bundle impact during refuelling and start-up of the reactor, and the acoustic pressure pulsations caused by the primary heat transport (PHT) pumps. The simulation is performed for a typical CANDU fuel bundle with 37 fuel elements. The clearances between the buttons of the inner fuel elements, and between the bearing pads of the outer fuel elements and the pressure tube were measured from an actual fuel bundle. Some variability among the measured clearance values was observed. Therefore, probability density functions of the measured clearance values were established and the simulation was performed for the probabilistic distribution of the clearance values. The contact between the fuel bundle and the pressure tube is modeled using pseudo-force contact method. The proposed modelling technique can be used in future CANDU reactors to avoid fuel and pressure tube fretting damage due to the aforementioned excitation mechanisms. (author)

Quantitative analysis of γ-oryzanol content in cold pressed rice bran oil by TLC-image analysis method.

Science.gov (United States)

Sakunpak, Apirak; Suksaeree, Jirapornchai; Monton, Chaowalit; Pathompak, Pathamaporn; Kraisintu, Krisana

2014-02-01

To develop and validate an image analysis method for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. TLC-densitometric and TLC-image analysis methods were developed, validated, and used for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. The results obtained by these two different quantification methods were compared by paired t-test. Both assays provided good linearity, accuracy, reproducibility and selectivity for determination of γ-oryzanol. The TLC-densitometric and TLC-image analysis methods provided a similar reproducibility, accuracy and selectivity for the quantitative determination of γ-oryzanol in cold pressed rice bran oil. A statistical comparison of the quantitative determinations of γ-oryzanol in samples did not show any statistically significant difference between TLC-densitometric and TLC-image analysis methods. As both methods were found to be equal, they therefore can be used for the determination of γ-oryzanol in cold pressed rice bran oil.

Quantitative scenario analysis of low and intermediate level radioactive repository

International Nuclear Information System (INIS)

Lee, Keon Jae; Lee, Sang Yoon; Park, Keon Baek; Song, Min Cheon; Lee, Ho Jin

1998-03-01

Derivation of hypothetical radioactive waste disposal facility os conducted through sub-component characteristic analysis and conceptual modeling. It is studied that quantitative analysis of constructed scenario in terms of annual effective dose equivalent. This study is sequentially conducted according to performance assessment of radioactive waste disposal facility such as : ground water flow analysis, source term analysis, ground water transport, surface water transport, dose and pathways. The routine program module such as VAM2D-PAGAN-GENII is used for quantitative scenario analysis. Detailed data used in this module are come from experimental data of Korean territory and default data given within this module. Is case of blank data for code execution, it is estimated through reasonable engineering sense

A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents

International Nuclear Information System (INIS)

Zhu, Xiaoming; Fu, Afu; Luo, Kathy Qian

2012-01-01

Highlights: ► An endothelial cell apoptosis assay using FRET-based biosensor was developed. ► The fluorescence of the cells changed from green to blue during apoptosis. ► This method was developed into a high-throughput assay in 96-well plates. ► This assay was applied to screen vascular disrupting agents. -- Abstract: In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening.

Assessment of fretting wear in Hanaro fuel

International Nuclear Information System (INIS)

Chae, Hee Taek; Lim, Kyeong Hwan; Kim, Hark Rho

1999-06-01

Since the first fuel loading on Feb. 1995, various zero-power tests were performed in HANARO and power ascending tests followed. After the initial fuel loading, Hanaro operation staffs inspected only two fuel bundles which were evaluated to have the highest power at the end of each cycle and they did not recognize anything peculiar in the inspected bundles. At the end of 1996, Hanaro staffs found severe wear damages in the fuel components. After that, the 4th cycle core was re-arranged with fresh fuels only to investigate wear phenomena on the fuel components. The fuel inspections have been performed 25 times periodically since the core re-configuration. In this report, fretting wear characteristics of the fuel assemblies were evaluated and summarized. Wear damages of the improved fuel assembly to resolve the wear problem were compared with those of the original fuel assembly. Based on the results of the fuel inspections, we suggest that fuel inspection need not be done for the first 60 pump operation days in order to reduce the potential of damage by a fuel handling error and an operator's burden of the fuel inspection. (author). 6 refs., 10 tabs., 5 figs

Microchromatography of hemoglobins. VIII. A general qualitative and quantitative method in plastic drinking straws and the quantitative analysis of Hb-F.

Science.gov (United States)

Schroeder, W A; Pace, L A

1978-03-01

The microchromatographic procedure for the quantitative analysis of the hemoglobin components in a hemolysate uses columns of DEAE-cellulose in a plastic drinking straw with a glycine-KCN-NaCl developer. Not only may the method be used for the quantitative analysis of Hb-F but also for the analysis of the varied components in mixtures of hemoglobins.

Quantitative genetic analysis of total glucosinolate, oil and protein ...

African Journals Online (AJOL)

Quantitative genetic analysis of total glucosinolate, oil and protein contents in Ethiopian mustard ( Brassica carinata A. Braun) ... Seeds were analyzed using HPLC (glucosinolates), NMR (oil) and NIRS (protein). Analyses of variance, Hayman's method of diallel analysis and a mixed linear model of genetic analysis were ...

Live-cell FRET imaging reveals clustering of the prion protein at the cell surface induced by infectious prions.

Science.gov (United States)

Tavares, Evandro; Macedo, Joana A; Paulo, Pedro M R; Tavares, Catarina; Lopes, Carlos; Melo, Eduardo P

2014-07-01

Prion diseases are associated to the conversion of the prion protein into a misfolded pathological isoform. The mechanism of propagation of protein misfolding by protein templating remains largely unknown. Neuroblastoma cells were transfected with constructs of the prion protein fused to both CFP-GPI-anchored and to YFP-GPI-anchored and directed to its cell membrane location. Live-cell FRET imaging between the prion protein fused to CFP or YFP was measured giving consistent values of 10±2%. This result was confirmed by fluorescence lifetime imaging microscopy and indicates intermolecular interactions between neighbor prion proteins. In particular, considering that a maximum FRET efficiency of 17±2% was determined from a positive control consisting of a fusion CFP-YFP-GPI-anchored. A stable cell clone expressing the two fusions containing the prion protein was also selected to minimize cell-to-cell variability. In both, stable and transiently transfected cells, the FRET efficiency consistently increased in the presence of infectious prions - from 4±1% to 7±1% in the stable clone and from 10±2% to 16±1% in transiently transfected cells. These results clearly reflect an increased clustering of the prion protein on the membrane in the presence of infectious prions, which was not observed in negative control using constructs without the prion protein and upon addition of non-infected brain. Our data corroborates the recent view that the primary site for prion conversion is the cell membrane. Since our fluorescent cell clone is not susceptible to propagate infectivity, we hypothesize that the initial event of prion infectivity might be the clustering of the GPI-anchored prion protein. Copyright © 2014 Elsevier B.V. All rights reserved.

Quantiprot - a Python package for quantitative analysis of protein sequences.

Science.gov (United States)

Konopka, Bogumił M; Marciniak, Marta; Dyrka, Witold

2017-07-17

The field of protein sequence analysis is dominated by tools rooted in substitution matrices and alignments. A complementary approach is provided by methods of quantitative characterization. A major advantage of the approach is that quantitative properties defines a multidimensional solution space, where sequences can be related to each other and differences can be meaningfully interpreted. Quantiprot is a software package in Python, which provides a simple and consistent interface to multiple methods for quantitative characterization of protein sequences. The package can be used to calculate dozens of characteristics directly from sequences or using physico-chemical properties of amino acids. Besides basic measures, Quantiprot performs quantitative analysis of recurrence and determinism in the sequence, calculates distribution of n-grams and computes the Zipf's law coefficient. We propose three main fields of application of the Quantiprot package. First, quantitative characteristics can be used in alignment-free similarity searches, and in clustering of large and/or divergent sequence sets. Second, a feature space defined by quantitative properties can be used in comparative studies of protein families and organisms. Third, the feature space can be used for evaluating generative models, where large number of sequences generated by the model can be compared to actually observed sequences.

The usefulness of 3D quantitative analysis with using MRI for measuring osteonecrosis of the femoral head

International Nuclear Information System (INIS)

Hwang, Ji Young; Lee, Sun Wha; Park, Youn Soo

2006-01-01

We wanted to evaluate the usefulness of MRI 3D quantitative analysis for measuring osteonecrosis of the femoral head in comparison with MRI 2D quantitative analysis and quantitative analysis of the specimen. For 3 months at our hospital, 14 femoral head specimens with osteonecrosis were obtained after total hip arthroplasty. The patients preoperative MRIs were retrospectively reviewed for quantitative analysis of the size of the necrosis. Each necrotic fraction of the femoral head was measured by 2D quantitative analysis with using mid-coronal and mid-sagittal MRIs, and by 3D quantitative analysis with using serial continuous coronal MRIs and 3D reconstruction software. The necrotic fraction of the specimen was physically measured by the fluid displacement method. The necrotic fraction according to MRI 2D or 3D quantitative analysis was compared with that of the specimen by using Spearman's correlation test. On the correlative analysis, the necrotic fraction by MRI 2D quantitative analysis and quantitative analysis of the specimen showed moderate correlation (r = 0.657); on the other hand, the necrotic fraction by MRI 3D quantitative analysis and quantitative analysis of the specimen demonstrated a strong correlation (r = 0.952) (ρ < 0.05). MRI 3D quantitative analysis was more accurate than 2D quantitative analysis using MRI for measuring osteonecrosis of the femoral head. Therefore, it may be useful for predicting the clinical outcome and deciding the proper treatment option

What Really Happens in Quantitative Group Research? Results of a Content Analysis of Recent Quantitative Research in "JSGW"

Science.gov (United States)

Boyle, Lauren H.; Whittaker, Tiffany A.; Eyal, Maytal; McCarthy, Christopher J.

2017-01-01

The authors conducted a content analysis on quantitative studies published in "The Journal for Specialists in Group Work" ("JSGW") between 2012 and 2015. This brief report provides a general overview of the current practices of quantitative group research in counseling. The following study characteristics are reported and…

Quantitative proteomic analysis of post-translational modifications of human histones

DEFF Research Database (Denmark)

Beck, Hans Christian; Nielsen, Eva C; Matthiesen, Rune

2006-01-01

, and H4 in a site-specific and dose-dependent manner. This unbiased analysis revealed that a relative increase in acetylated peptide from the histone variants H2A, H2B, and H4 was accompanied by a relative decrease of dimethylated Lys(57) from histone H2B. The dose-response results obtained...... by quantitative proteomics of histones from HDACi-treated cells were consistent with Western blot analysis of histone acetylation, cytotoxicity, and dose-dependent expression profiles of p21 and cyclin A2. This demonstrates that mass spectrometry-based quantitative proteomic analysis of post-translational...

Data from quantitative label free proteomics analysis of rat spleen.

Science.gov (United States)

Dudekula, Khadar; Le Bihan, Thierry

2016-09-01

The dataset presented in this work has been obtained using a label-free quantitative proteomic analysis of rat spleen. A robust method for extraction of proteins from rat spleen tissue and LC-MS-MS analysis was developed using a urea and SDS-based buffer. Different fractionation methods were compared. A total of 3484 different proteins were identified from the pool of all experiments run in this study (a total of 2460 proteins with at least two peptides). A total of 1822 proteins were identified from nine non-fractionated pulse gels, 2288 proteins and 2864 proteins were identified by SDS-PAGE fractionation into three and five fractions respectively. The proteomics data are deposited in ProteomeXchange Consortium via PRIDE PXD003520, Progenesis and Maxquant output are presented in the supported information. The generated list of proteins under different regimes of fractionation allow assessing the nature of the identified proteins; variability in the quantitative analysis associated with the different sampling strategy and allow defining a proper number of replicates for future quantitative analysis.

Mitochondrial intermediate peptidase: Expression in Escherichia coli and improvement of its enzymatic activity detection with FRET substrates

International Nuclear Information System (INIS)

Marcondes, Marcelo F.; Torquato, Ricardo J.S.; Assis, Diego M.; Juliano, Maria A.; Hayashi, Mirian A.F.; Oliveira, Vitor

2010-01-01

In the present study, soluble, functionally-active, recombinant human mitochondrial intermediate peptidase (hMIP), a mitochondrial metalloendoprotease, was expressed in a prokaryotic system. The hMIP fusion protein, with a poly-His-tag (6x His), was obtained by cloning the coding region of hMIP cDNA into the pET-28a expression vector, which was then used to transform Escherichia coli BL21 (DE3) pLysS. After isolation and purification of the fusion protein by affinity chromatography using Ni-Sepharose resin, the protein was purified further using ion exchange chromatography with a Hi-trap resource Q column. The recombinant hMIP was characterized by Western blotting using three distinct antibodies, circular dichroism, and enzymatic assays that used the first FRET substrates developed for MIP and a series of protease inhibitors. The successful expression of enzymatically-active hMIP in addition to the FRET substrates will contribute greatly to the determination of substrate specificity of this protease and to the development of specific inhibitors that are essential for a better understanding of the role of this protease in mitochondrial functioning.

Quantitative analysis of some brands of chloroquine tablets ...

African Journals Online (AJOL)

Quantitative analysis of some brands of chloroquine tablets marketed in Maiduguri using spectrophotometric ... and compared with that of the standard, wavelength of maximum absorbance at 331nm for chloroquine. ... HOW TO USE AJOL.

Accurate quantitative XRD phase analysis of cement clinkers

International Nuclear Information System (INIS)

Kern, A.

2002-01-01

Full text: Knowledge about the absolute phase abundance in cement clinkers is a requirement for both, research and quality control. Traditionally, quantitative analysis of cement clinkers has been carried out by theoretical normative calculation from chemical analysis using the so-called Bogue method or by optical microscopy. Therefore chemical analysis, mostly performed by X-ray fluorescence (XRF), forms the basis of cement plan control by providing information for proportioning raw materials, adjusting kiln and burning conditions, as well as cement mill feed proportioning. In addition, XRF is of highest importance with respect to the environmentally relevant control of waste recovery raw materials and alternative fuels, as well as filters, plants and sewage. However, the performance of clinkers and cements is governed by the mineralogy and not the elemental composition, and the deficiencies and inherent errors of Bogue as well as microscopic point counting are well known. With XRD and Rietveld analysis a full quantitative analysis of cement clinkers can be performed providing detailed mineralogical information about the product. Until recently several disadvantages prevented the frequent application of the Rietveld method in the cement industry. As the measurement of a full pattern is required, extended measurement times made an integration of this method into existing automation environments difficult. In addition, several drawbacks of existing Rietveld software such as complexity, low performance and severe numerical instability were prohibitive for automated use. The latest developments of on-line instrumentation, as well as dedicated Rietveld software for quantitative phase analysis (TOPAS), now make a decisive breakthrough possible. TOPAS not only allows the analysis of extremely complex phase mixtures in the shortest time possible, but also a fully automated online phase analysis for production control and quality management, free of any human interaction

Global scaling for semi-quantitative analysis in FP-CIT SPECT.

Science.gov (United States)

Kupitz, D; Apostolova, I; Lange, C; Ulrich, G; Amthauer, H; Brenner, W; Buchert, R

2014-01-01

Semi-quantitative characterization of dopamine transporter availability from single photon emission computed tomography (SPECT) with 123I-ioflupane (FP-CIT) is based on uptake ratios relative to a reference region. The aim of this study was to evaluate the whole brain as reference region for semi-quantitative analysis of FP-CIT SPECT. The rationale was that this might reduce statistical noise associated with the estimation of non-displaceable FP-CIT uptake. 150 FP-CIT SPECTs were categorized as neurodegenerative or non-neurodegenerative by an expert. Semi-quantitative analysis of specific binding ratios (SBR) was performed with a custom-made tool based on the Statistical Parametric Mapping software package using predefined regions of interest (ROIs) in the anatomical space of the Montreal Neurological Institute. The following reference regions were compared: predefined ROIs for frontal and occipital lobe and whole brain (without striata, thalamus and brainstem). Tracer uptake in the reference region was characterized by the mean, median or 75th percentile of its voxel intensities. The area (AUC) under the receiver operating characteristic curve was used as performance measure. The highest AUC of 0.973 was achieved by the SBR of the putamen with the 75th percentile in the whole brain as reference. The lowest AUC for the putamen SBR of 0.937 was obtained with the mean in the frontal lobe as reference. We recommend the 75th percentile in the whole brain as reference for semi-quantitative analysis in FP-CIT SPECT. This combination provided the best agreement of the semi-quantitative analysis with visual evaluation of the SPECT images by an expert and, therefore, is appropriate to support less experienced physicians.

Quantitative possibility analysis. Present status in ESCA

International Nuclear Information System (INIS)

Brion, D.

1981-01-01

A short review of the recent developments in quantification of X-ray photoelectron spectroscopy or ESCA is presented. The basic equations are reminded. Each involved parameter (photoionisation, inelastic mean free paths, 'response function' of the instruments, intensity measurement) is separately discussed in relation with the accuracy and the precision of the method. Other topics are considered such as roughness, surface contamination, matrix effect and inhomogeneous composition. Some aspects of the quantitative ESCA analysis and AES analysis are compared [fr

Uncertainties in elemental quantitative analysis by PIXE

International Nuclear Information System (INIS)

Montenegro, E.C.; Baptista, G.B.; Paschoa, A.S.; Barros Leite, C.V.

1979-01-01

The effects of the degree of non-uniformity of the particle beam, matrix composition and matrix thickness in a quantitative elemental analysis by particle induced X-ray emission (PIXE) are discussed and a criterion to evaluate the resulting degree of uncertainty in the mass determination by this method is established. (Auth.)

Quantitative analysis of γ-oryzanol content in cold pressed rice bran oil by TLC-image analysis method

OpenAIRE

Sakunpak, Apirak; Suksaeree, Jirapornchai; Monton, Chaowalit; Pathompak, Pathamaporn; Kraisintu, Krisana

2014-01-01

Objective: To develop and validate an image analysis method for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. Methods: TLC-densitometric and TLC-image analysis methods were developed, validated, and used for quantitative analysis of γ-oryzanol in cold pressed rice bran oil. The results obtained by these two different quantification methods were compared by paired t-test. Results: Both assays provided good linearity, accuracy, reproducibility and selectivity for dete...

Evaluation of breast lesions by contrast enhanced ultrasound: Qualitative and quantitative analysis

International Nuclear Information System (INIS)

Wan Caifeng; Du Jing; Fang Hua; Li Fenghua; Wang Lin

2012-01-01

Objective: To evaluate and compare the diagnostic performance of qualitative, quantitative and combined analysis for characterization of breast lesions in contrast enhanced ultrasound (CEUS), with histological results used as the reference standard. Methods: Ninety-one patients with 91 breast lesions BI-RADS 3–5 at US or mammography underwent CEUS. All lesions underwent qualitative and quantitative enhancement evaluation. Receiver operating characteristic (ROC) curve analysis was performed to evaluate the diagnostic performance of different analytical method for discrimination between benign and malignant breast lesions. Results: Histopathologic analysis of the 91 lesions revealed 44 benign and 47 malignant. For qualitative analysis, benign and malignant lesions differ significantly in enhancement patterns (p z1 ), 0.768 (A z2 ) and 0.926(A z3 ) respectively. The values of A z1 and A z3 were significantly higher than that for A z2 (p = 0.024 and p = 0.008, respectively). But there was no significant difference between the values of A z1 and A z3 (p = 0.625). Conclusions: The diagnostic performance of qualitative and combined analysis was significantly higher than that for quantitative analysis. Although quantitative analysis has the potential to differentiate benign from malignant lesions, it has not yet improved the final diagnostic accuracy.

Determination of cDNA encoding BCR/ABL fusion gene in patients with chronic myelogenous leukemia using a novel FRET-based quantum dots-DNA nanosensor.

Science.gov (United States)

Shamsipur, Mojtaba; Nasirian, Vahid; Barati, Ali; Mansouri, Kamran; Vaisi-Raygani, Asad; Kashanian, Soheila

2017-05-08

In the present study, we developed a sensitive method based on fluorescence resonance energy transfer (FRET) for the determination of the BCR/ABL fusion gene, which is used as a biomarker to confirm the clinical diagnosis of both chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL). For this purpose, CdTe quantum dots (QDs) were conjugated to amino-modified 18-mer oligonucleotide ((N)DNA) to form the QDs-(N)DNA nanosensor. In the presence of methylene blue (MB) as an intercalator, the hybridization of QDs-(N)DNA with the target BCR/ABL fusion gene (complementary DNA), brings the MB (acceptor) at close proximity of the QDs (donor), leading to FRET upon photoexcitation of the QDs. The enhancement in the emission intensity of MB was used to follow up the hybridization, which was linearly proportional to concentration of the target complementary DNA in a range from 1.0 × 10 -9 to 1.25 × 10 -7  M. The detection limit of the proposed method was obtained to be 1.5 × 10 -10  M. Finally, the feasibility and selectivity of the proposed nanosensor was evaluated by the analysis of derived nucleotides from both mismatched sequences and clinical samples of patients with leukemia as real samples. Copyright © 2017 Elsevier B.V. All rights reserved.

New Approach to Quantitative Analysis by Laser-induced Breakdown Spectroscopy

International Nuclear Information System (INIS)

Lee, D. H.; Kim, T. H.; Yun, J. I.; Jung, E. C.

2009-01-01

Laser-induced breakdown spectroscopy (LIBS) has been studied as the technique of choice in some particular situations like screening, in situ measurement, process monitoring, hostile environments, etc. Especially, LIBS can fulfill the qualitative and quantitative analysis for radioactive high level waste (HLW) glass in restricted experimental conditions. Several ways have been suggested to get quantitative information from LIBS. The one approach is to use the absolute intensities of each element. The other approach is to use the elemental emission intensities relative to the intensity of the internal standard element whose concentration is known already in the specimen. But these methods are not applicable to unknown samples. In the present work, we introduce new approach to LIBS quantitative analysis by using H α (656.28 nm) emission line as external standard

Time-Resolved Analysis of a Highly Sensitive Förster Resonance Energy Transfer Immunoassay Using Terbium Complexes as Donors and Quantum Dots as Acceptors

Directory of Open Access Journals (Sweden)

Niko Hildebrandt

2007-01-01

Full Text Available CdSe/ZnS core/shell quantum dots (QDs are used as efficient Förster Resonance Energy Transfer (FRET acceptors in a time-resolved immunoassays with Tb complexes as donors providing a long-lived luminescence decay. A detailed decay time analysis of the FRET process is presented. QD FRET sensitization is evidenced by a more than 1000-fold increase of the QD luminescence decay time reaching ca. 0.5 milliseconds, the same value to which the Tb donor decay time is quenched due to FRET to the QD acceptors. The FRET system has an extremely large Förster radius of approx. 100 Å and more than 70% FRET efficiency with a mean donor-acceptor distance of ca. 84 Å, confirming the applied biotin-streptavidin binding system. Time-resolved measurement allows for suppression of short-lived emission due to background fluorescence and directly excited QDs. By this means a detection limit of 18 attomol QDs within the immunoassay is accomplished, an improvement of more than two orders of magnitude compared to commercial systems.

Multicomponent quantitative spectroscopic analysis without reference substances based on ICA modelling.

Science.gov (United States)

Monakhova, Yulia B; Mushtakova, Svetlana P

2017-05-01

A fast and reliable spectroscopic method for multicomponent quantitative analysis of targeted compounds with overlapping signals in complex mixtures has been established. The innovative analytical approach is based on the preliminary chemometric extraction of qualitative and quantitative information from UV-vis and IR spectral profiles of a calibration system using independent component analysis (ICA). Using this quantitative model and ICA resolution results of spectral profiling of "unknown" model mixtures, the absolute analyte concentrations in multicomponent mixtures and authentic samples were then calculated without reference solutions. Good recoveries generally between 95% and 105% were obtained. The method can be applied to any spectroscopic data that obey the Beer-Lambert-Bouguer law. The proposed method was tested on analysis of vitamins and caffeine in energy drinks and aromatic hydrocarbons in motor fuel with 10% error. The results demonstrated that the proposed method is a promising tool for rapid simultaneous multicomponent analysis in the case of spectral overlap and the absence/inaccessibility of reference materials.

Application of phasor plot and autofluorescence correction for study of heterogeneous cell population

Science.gov (United States)

Szmacinski, Henryk; Toshchakov, Vladimir; Lakowicz, Joseph R.

2014-01-01

Abstract. Protein-protein interactions in cells are often studied using fluorescence resonance energy transfer (FRET) phenomenon by fluorescence lifetime imaging microscopy (FLIM). Here, we demonstrate approaches to the quantitative analysis of FRET in cell population in a case complicated by a highly heterogeneous donor expression, multiexponential donor lifetime, large contribution of cell autofluorescence, and significant presence of unquenched donor molecules that do not interact with the acceptor due to low affinity of donor-acceptor binding. We applied a multifrequency phasor plot to visualize FRET FLIM data, developed a method for lifetime background correction, and performed a detailed time-resolved analysis using a biexponential model. These approaches were applied to study the interaction between the Toll Interleukin-1 receptor (TIR) domain of Toll-like receptor 4 (TLR4) and the decoy peptide 4BB. TLR4 was fused to Cerulean fluorescent protein (Cer) and 4BB peptide was labeled with Bodipy TMRX (BTX). Phasor displays for multifrequency FLIM data are presented. The analytical procedure for lifetime background correction is described and the effect of correction on FLIM data is demonstrated. The absolute FRET efficiency was determined based on the phasor plot display and multifrequency FLIM data analysis. The binding affinity between TLR4-Cer (donor) and decoy peptide 4BB-BTX (acceptor) was estimated in a heterogeneous HeLa cell population. PMID:24770662

Dual Mechanism of an Intramolecular Charge Transfer (ICT)-FRET-Based Fluorescent Probe for the Selective Detection of Hydrogen Peroxide.

Science.gov (United States)

Liang, Xiao; Xu, Xiaoyi; Qiao, Dan; Yin, Zheng; Shang, Luqing

2017-12-14

A dual-mechanism intramolecular charge transfer (ICT)-FRET fluorescent probe for the selective detection of H 2 O 2 in living cells has been designed and synthesized. This probe used a coumarin-naphthalimide hybrid as the FRET platform and a boronate moiety as the recognition group. Upon the addition of H 2 O 2 , the probe exhibited a redshifted (73 nm) fluorescence emission, and the ratio of fluorescence intensities at λ=558 and 485 nm (F 558 /F 485 ) shifted notably (up to 100-fold). Moreover, there was a good linearity (R 2 =0.9911) between the ratio and concentration of H 2 O 2 in the range of 0 to 60 μm, with a limit of detection of 0.28 μm (signal to noise ratio (S/N)=3). This probe could also detect enzymatically generated H 2 O 2 . Importantly, it could be used to visualize endogenous H 2 O 2 produced by stimulation from epidermal growth factor. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fiber Fabry-Perot Force Sensor with Small Volume and High Performance for Assessing Fretting Damage of Steam Generator Tubes.

Science.gov (United States)

Huang, Peijian; Wang, Ning; Li, Junying; Zhu, Yong; Zhang, Jie

2017-12-13

Measuring the radial collision force between the steam generator tube (SGT) and the tube support plate (TSP) is essential to assess the fretting damage of the SGT. In order to measure the radial collision force, a novel miniaturized force sensor based on fiber Fabry-Perot (F-P) was designed, and the principle and characteristics of the sensor were analyzed in detail. Then, the F-P force sensor was successfully fabricated and calibrated, and the overall dimensions of the encapsulated fiber F-P sensor were 17 mm × 5 mm × 3 mm (L × W × H). The sensor works well in humid, high pressure (10 MPa), high temperature (350 °C), and vibration (40 kHz) environments. Finally, the F-P force sensors were installed in a 1:1 steam generator test loop, and the radial collision force signals between the SGT and the TSP were obtained. The experiments indicated that the F-P sensor with small volume and high performance could help in assessing the fretting damage of the steam generator tubes.

A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles

International Nuclear Information System (INIS)

Li, Yaqi; Sun, Li; Qian, Jing; Wang, Chengke; Liu, Qian; Han, En; Hao, Nan; Zhang, Liuping; Cai, Jianrong; Wang, Kun

2016-01-01

In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1–500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection. - Highlights: • Both NGQDs and AgNPs were selected as the novel FRET donor-acceptor pairs. • The proposed homogeneous FRET assay was developed for CaMV35S detection. • The resulting method could identify 0.5% containing transgenic soybean sample. • This assay was inexpensive, simple and highly sensitive.

A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Li, Yaqi; Sun, Li [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Qian, Jing [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China); Wang, Chengke [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Liu, Qian [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China); Han, En [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Hao, Nan [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China); Zhang, Liuping [Sinograin Zhenjiang Grains & Oils Quality Testing Center Co., Ltd., Zhenjiang, 212013 (China); Cai, Jianrong, E-mail: jrcai@ujs.edu.cn [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Wang, Kun, E-mail: wangkun@ujs.edu.cn [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China)

2016-12-15

In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1–500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection. - Highlights: • Both NGQDs and AgNPs were selected as the novel FRET donor-acceptor pairs. • The proposed homogeneous FRET assay was developed for CaMV35S detection. • The resulting method could identify 0.5% containing transgenic soybean sample. • This assay was inexpensive, simple and highly sensitive.

ImatraNMR: Novel software for batch integration and analysis of quantitative NMR spectra

Science.gov (United States)

Mäkelä, A. V.; Heikkilä, O.; Kilpeläinen, I.; Heikkinen, S.

2011-08-01

Quantitative NMR spectroscopy is a useful and important tool for analysis of various mixtures. Recently, in addition of traditional quantitative 1D 1H and 13C NMR methods, a variety of pulse sequences aimed for quantitative or semiquantitative analysis have been developed. To obtain actual usable results from quantitative spectra, they must be processed and analyzed with suitable software. Currently, there are many processing packages available from spectrometer manufacturers and third party developers, and most of them are capable of analyzing and integration of quantitative spectra. However, they are mainly aimed for processing single or few spectra, and are slow and difficult to use when large numbers of spectra and signals are being analyzed, even when using pre-saved integration areas or custom scripting features. In this article, we present a novel software, ImatraNMR, designed for batch analysis of quantitative spectra. In addition to capability of analyzing large number of spectra, it provides results in text and CSV formats, allowing further data-analysis using spreadsheet programs or general analysis programs, such as Matlab. The software is written with Java, and thus it should run in any platform capable of providing Java Runtime Environment version 1.6 or newer, however, currently it has only been tested with Windows and Linux (Ubuntu 10.04). The software is free for non-commercial use, and is provided with source code upon request.

Quantitative subsurface analysis using frequency modulated thermal wave imaging

Science.gov (United States)

Subhani, S. K.; Suresh, B.; Ghali, V. S.

2018-01-01

Quantitative depth analysis of the anomaly with an enhanced depth resolution is a challenging task towards the estimation of depth of the subsurface anomaly using thermography. Frequency modulated thermal wave imaging introduced earlier provides a complete depth scanning of the object by stimulating it with a suitable band of frequencies and further analyzing the subsequent thermal response using a suitable post processing approach to resolve subsurface details. But conventional Fourier transform based methods used for post processing unscramble the frequencies with a limited frequency resolution and contribute for a finite depth resolution. Spectral zooming provided by chirp z transform facilitates enhanced frequency resolution which can further improves the depth resolution to axially explore finest subsurface features. Quantitative depth analysis with this augmented depth resolution is proposed to provide a closest estimate to the actual depth of subsurface anomaly. This manuscript experimentally validates this enhanced depth resolution using non stationary thermal wave imaging and offers an ever first and unique solution for quantitative depth estimation in frequency modulated thermal wave imaging.

FRET measurements of kinesin neck orientation reveal a structural basis for processivity and asymmetry.

Science.gov (United States)

Martin, Douglas S; Fathi, Reza; Mitchison, Timothy J; Gelles, Jeff

2010-03-23

As the smallest and simplest motor enzymes, kinesins have served as the prototype for understanding the relationship between protein structure and mechanochemical function of enzymes in this class. Conventional kinesin (kinesin-1) is a motor enzyme that transports cargo toward the plus end of microtubules by a processive, asymmetric hand-over-hand mechanism. The coiled-coil neck domain, which connects the two kinesin motor domains, contributes to kinesin processivity (the ability to take many steps in a row) and is proposed to be a key determinant of the asymmetry in the kinesin mechanism. While previous studies have defined the orientation and position of microtubule-bound kinesin motor domains, the disposition of the neck coiled-coil remains uncertain. We determined the neck coiled-coil orientation using a multidonor fluorescence resonance energy transfer (FRET) technique to measure distances between microtubules and bound kinesin molecules. Microtubules were labeled with a new fluorescent taxol donor, TAMRA-X-taxol, and kinesin derivatives with an acceptor fluorophore attached at positions on the motor and neck coiled-coil domains were used to reconstruct the positions and orientations of the domains. FRET measurements to positions on the motor domain were largely consistent with the domain orientation determined in previous studies, validating the technique. Measurements to positions on the neck coiled-coil were inconsistent with a radial orientation and instead demonstrated that the neck coiled-coil is parallel to the microtubule surface. The measured orientation provides a structural explanation for how neck surface residues enhance processivity and suggests a simple hypothesis for the origin of kinesin step asymmetry and "limping."

A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles.

Science.gov (United States)

Li, Yaqi; Sun, Li; Qian, Jing; Wang, Chengke; Liu, Qian; Han, En; Hao, Nan; Zhang, Liuping; Cai, Jianrong; Wang, Kun

2016-12-15

In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1-500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection. Copyright © 2016 Elsevier B.V. All rights reserved.

Quantitative analysis by nuclear magnetic resonance spectroscopy

Energy Technology Data Exchange (ETDEWEB)

Wainai, T; Mashimo, K [Nihon Univ., Tokyo. Coll. of Science and Engineering

1976-04-01

Recent papers on the practical quantitative analysis by nuclear magnetic resonance spectroscopy (NMR) are reviewed. Specifically, the determination of moisture in liquid N/sub 2/O/sub 4/ as an oxidizing agent for rocket propulsion, the analysis of hydroperoxides, the quantitative analysis using a shift reagent, the analysis of aromatic sulfonates, and the determination of acids and bases are reviewed. Attention is paid to the accuracy. The sweeping velocity and RF level in addition to the other factors must be on the optimal condition to eliminate the errors, particularly when computation is made with a machine. Higher sweeping velocity is preferable in view of S/N ratio, but it may be limited to 30 Hz/s. The relative error in the measurement of area is generally 1%, but when those of dilute concentration and integrated, the error will become smaller by one digit. If impurities are treated carefully, the water content on N/sub 2/O/sub 4/ can be determined with accuracy of about 0.002%. The comparison method between peak heights is as accurate as that between areas, when the uniformity of magnetic field and T/sub 2/ are not questionable. In the case of chemical shift movable due to content, the substance can be determined by the position of the chemical shift. Oil and water contents in rape-seed, peanuts, and sunflower-seed are determined by measuring T/sub 1/ with 90 deg pulses.

The Conformational Dynamics of Cas9 Governing DNA Cleavage Are Revealed by Single-Molecule FRET

Directory of Open Access Journals (Sweden)

Mengyi Yang

2018-01-01

Full Text Available Summary: Off-target binding and cleavage by Cas9 pose major challenges in its application. How the conformational dynamics of Cas9 govern its nuclease activity under on- and off-target conditions remains largely unknown. Here, using intra-molecular single-molecule fluorescence resonance energy transfer measurements, we revealed that Cas9 in apo, sgRNA-bound, and dsDNA/sgRNA-bound forms spontaneously transits among three major conformational states, mainly reflecting significant conformational mobility of the catalytic HNH domain. We also uncovered surprising long-range allosteric communication between the HNH domain and the RNA/DNA heteroduplex at the PAM-distal end to ensure correct positioning of the catalytic site, which demonstrated that a unique proofreading mechanism served as the last checkpoint before DNA cleavage. Several Cas9 residues were likely to mediate the allosteric communication and proofreading step. Modulating interactions between Cas9 and heteroduplex at the PAM-distal end by introducing mutations on these sites provides an alternative route to improve and optimize the CRISPR/Cas9 toolbox. : Yang et al. revealed significant conformational dynamics of Cas9 at global and local scales using single-molecule FRET. They uncovered surprising long-range allosteric communication between the HNH nuclease domain and the RNA/DNA heteroduplex at the PAM-distal end that serves as a proofreading checkpoint to govern the nuclease activity and specificity of Cas9. Keywords: CRISPR, Cas9, single-molecule, FRET, conformational dynamics, proofreading, off-target, allosteric communication, genome editing

Stable Isotope Quantitative N-Glycan Analysis by Liquid Separation Techniques and Mass Spectrometry.

Science.gov (United States)

Mittermayr, Stefan; Albrecht, Simone; Váradi, Csaba; Millán-Martín, Silvia; Bones, Jonathan

2017-01-01

Liquid phase separation analysis and subsequent quantitation remains a challenging task for protein-derived oligosaccharides due to their inherent structural complexity and diversity. Incomplete resolution or co-detection of multiple glycan species complicates peak area-based quantitation and associated statistical analysis when optical detection methods are used. The approach outlined herein describes the utilization of stable isotope variants of commonly used fluorescent tags that allow for mass-based glycan identification and relative quantitation following separation by liquid chromatography (LC) or capillary electrophoresis (CE). Comparability assessment of glycoprotein-derived oligosaccharides is performed by derivatization with commercially available isotope variants of 2-aminobenzoic acid or aniline and analysis by LC- and CE-mass spectrometry. Quantitative information is attained from the extracted ion chromatogram/electropherogram ratios generated from the light and heavy isotope clusters.

Quantitative analysis of myocardial tissue with digital autofluorescence microscopy

DEFF Research Database (Denmark)

Jensen, Thomas; Holten-Rossing, Henrik; Svendsen, Ida M H

2016-01-01

to that of hematoxylin and eosin staining in conventional pathology. This study presents an automated fluorescence-based microscopy approach providing highly detailed morphological data from unstained microsections. This data may provide a basic histological starting point from which further digital analysis including...... staining may benefit. METHODS: This study explores the inherent tissue fluorescence, also known as autofluorescence, as a mean to quantitate cardiac tissue components in histological microsections. Data acquisition using a commercially available whole slide scanner and an image-based quantitation algorithm......BACKGROUND: The opportunity offered by whole slide scanners of automated histological analysis implies an ever increasing importance of digital pathology. To go beyond the importance of conventional pathology, however, digital pathology may need a basic histological starting point similar...

Quantitative analysis of target components by comprehensive two-dimensional gas chromatography

NARCIS (Netherlands)

Mispelaar, V.G. van; Tas, A.C.; Smilde, A.K.; Schoenmakers, P.J.; Asten, A.C. van

2003-01-01

Quantitative analysis using comprehensive two-dimensional (2D) gas chromatography (GC) is still rarely reported. This is largely due to a lack of suitable software. The objective of the present study is to generate quantitative results from a large GC x GC data set, consisting of 32 chromatograms.

Role of image analysis in quantitative characterisation of nuclear fuel materials

International Nuclear Information System (INIS)

Dubey, J.N.; Rao, T.S.; Pandey, V.D.; Majumdar, S.

2005-01-01

Image analysis is one of the important techniques, widely used for materials characterization. It provides the quantitative estimation of the microstructural features present in the material. This information is very much valuable for finding out the criteria for taking up the fuel for high burn up. Radiometallurgy Division has been carrying out development and fabrication of plutonium related fuels for different type of reactors viz. Purnima, Fast Breeder Test Reactor (FBTR), Prototype Fast Breeder Reactor (PFBR), Boiling Water Reactor (BWR), Advanced Heavy Water Reactor (AHWR), Pressurised Heavy Water Reactor (PHWR) and KAMINI Reactor. Image analysis has been carried out on microstructures of PHWR, AHWR, FBTR and KAMINI fuels. Samples were prepared as per standard ASTM metallographic procedure. Digital images of the microstructure of these specimens were obtained using CCD camera, attached to the optical microscope. These images are stores on computer and used for detection and analysis of features of interest with image analysis software. Quantitative image analysis technique has been standardised and used for finding put type of the porosity, its size, shape and distribution in the above sintered oxide and carbide fuels. This technique has also been used for quantitative estimation of different phases present in KAMINI fuel. Image analysis results have been summarised and presented in this paper. (author)

Micro photometer's automation for quantitative spectrograph analysis

International Nuclear Information System (INIS)

Gutierrez E, C.Y.A.

1996-01-01

A Microphotometer is used to increase the sharpness of dark spectral lines. Analyzing these lines one sample content and its concentration could be determined and the analysis is known as Quantitative Spectrographic Analysis. The Quantitative Spectrographic Analysis is carried out in 3 steps, as follows. 1. Emulsion calibration. This consists of gauging a photographic emulsion, to determine the intensity variations in terms of the incident radiation. For the procedure of emulsion calibration an adjustment with square minimum to the data obtained is applied to obtain a graph. It is possible to determine the density of dark spectral line against the incident light intensity shown by the microphotometer. 2. Working curves. The values of known concentration of an element against incident light intensity are plotted. Since the sample contains several elements, it is necessary to find a work curve for each one of them. 3. Analytical results. The calibration curve and working curves are compared and the concentration of the studied element is determined. The automatic data acquisition, calculation and obtaining of resulting, is done by means of a computer (PC) and a computer program. The conditioning signal circuits have the function of delivering TTL levels (Transistor Transistor Logic) to make the communication between the microphotometer and the computer possible. Data calculation is done using a computer programm

[Rapid analysis of suppositories by quantitative 1H NMR spectroscopy].

Science.gov (United States)

Abramovich, R A; Kovaleva, S A; Goriainov, S V; Vorob'ev, A N; Kalabin, G A

2012-01-01

Rapid analysis of suppositories with ibuprofen and arbidol by quantitative 1H NMR spectroscopy was performed. Optimal conditions for the analysis were developed. The results are useful for design of rapid methods for quality control of suppositories with different components

Quantitative EDXS analysis of organic materials using the ζ-factor method

International Nuclear Information System (INIS)

Fladischer, Stefanie; Grogger, Werner

2014-01-01

In this study we successfully applied the ζ-factor method to perform quantitative X-ray analysis of organic thin films consisting of light elements. With its ability to intrinsically correct for X-ray absorption, this method significantly improved the quality of the quantification as well as the accuracy of the results compared to conventional techniques in particular regarding the quantification of light elements. We describe in detail the process of determining sensitivity factors (ζ-factors) using a single standard specimen and the involved parameter optimization for the estimation of ζ-factors for elements not contained in the standard. The ζ-factor method was then applied to perform quantitative analysis of organic semiconducting materials frequently used in organic electronics. Finally, the results were verified and discussed concerning validity and accuracy. - Highlights: • The ζ-factor method is used for quantitative EDXS analysis of light elements. • We describe the process of determining ζ-factors from a single standard in detail. • Organic semiconducting materials are successfully quantified

Quantitative analysis by computer controlled X-ray fluorescence spectrometer

International Nuclear Information System (INIS)

Balasubramanian, T.V.; Angelo, P.C.

1981-01-01

X-ray fluorescence spectroscopy has become a widely accepted method in the metallurgical field for analysis of both minor and major elements. As encountered in many other analytical techniques, the problem of matrix effect generally known as the interelemental effects is to be dealt with effectively in order to make the analysis accurate. There are several methods by which the effects of matrix on the analyte are minimised or corrected for and the mathematical correction is one among them. In this method the characteristic secondary X-ray intensities are measured from standard samples and correction coefficients. If any, for interelemental effects are evaluated by mathematical calculations. This paper describes attempts to evaluate the correction coefficients for interelemental effects by multiple linear regression programmes using a computer for the quantitative analysis of stainless steel and a nickel base cast alloy. The quantitative results obtained using this method for a standard stainless steel sample are compared with the given certified values. (author)

PIQMIe: A web server for semi-quantitative proteomics data management and analysis

NARCIS (Netherlands)

A. Kuzniar (Arnold); R. Kanaar (Roland)

2014-01-01

textabstractWe present the Proteomics Identifications and Quantitations Data Management and Integration Service or PIQMIe that aids in reliable and scalable data management, analysis and visualization of semi-quantitative mass spectrometry based proteomics experiments. PIQMIe readily integrates

Issues in Quantitative Analysis of Ultraviolet Imager (UV) Data: Airglow

Science.gov (United States)

Germany, G. A.; Richards, P. G.; Spann, J. F.; Brittnacher, M. J.; Parks, G. K.

1999-01-01

The GGS Ultraviolet Imager (UVI) has proven to be especially valuable in correlative substorm, auroral morphology, and extended statistical studies of the auroral regions. Such studies are based on knowledge of the location, spatial, and temporal behavior of auroral emissions. More quantitative studies, based on absolute radiometric intensities from UVI images, require a more intimate knowledge of the instrument behavior and data processing requirements and are inherently more difficult than studies based on relative knowledge of the oval location. In this study, UVI airglow observations are analyzed and compared with model predictions to illustrate issues that arise in quantitative analysis of UVI images. These issues include instrument calibration, long term changes in sensitivity, and imager flat field response as well as proper background correction. Airglow emissions are chosen for this study because of their relatively straightforward modeling requirements and because of their implications for thermospheric compositional studies. The analysis issues discussed here, however, are identical to those faced in quantitative auroral studies.

Critical appraisal of semi-quantitative analysis of 2-deoxyglucose autoradiograms

Energy Technology Data Exchange (ETDEWEB)

Kelly, P T; McCulloch, J [Glasgow Univ. (UK)

1983-06-13

Semi-quantitative analysis (e.g. optical density ratios) of (/sup 14/C)2-deoxyglucose autoradiograms is widely used in neuroscience research. The authors demonstrate that a fixed ratio of /sup 14/C-concentrations in the CNS does not yield a constant optical density ratio but is dependent upon the exposure time in the preparation of the autoradiograms and the absolute amounts of /sup 14/C from which the concentration ratio is derived. The failure of a fixed glucose utilization ratio to result in a constant optical density ratio represents a major interpretative difficulty in investigations where only semi-quantitative analysis of (/sup 14/C)2-deoxyglucose autoradiograms is undertaken.

Quantitative-genetic analysis of wing form and bilateral asymmetry ...

Indian Academy of Sciences (India)

Unknown

lines; Procrustes analysis; wing shape; wing size. ... Models of stochastic gene expression pre- dict that intrinsic noise ... Quantitative parameters of wing size and shape asymmetries ..... the residuals of a regression on centroid size produced.

Quantitative Analysis of Complex Tropical Forest Stands: A Review ...

African Journals Online (AJOL)

FIRST LADY

The importance of data analysis in quantitative assessment of natural resources .... Data collection design is an important process in complex forest statistical ... Ideally, the sample size should be equal among groups and sufficiently large.

Transportation and quantitative analysis of socio-economic development of relations

Science.gov (United States)

Chen, Yun

2017-12-01

Transportation has a close relationship with socio-economic. This article selects the indicators which can measure the development of transportation and socio-economic, using the method of correlation analysis, regression analysis, intensity of transportation analysis and transport elastic analysis, to analyze the relationship between them quantitatively, so that it has the fact guiding sense in the national development planning for the future.

Quantitative high-resolution genomic analysis of single cancer cells.

Science.gov (United States)

Hannemann, Juliane; Meyer-Staeckling, Sönke; Kemming, Dirk; Alpers, Iris; Joosse, Simon A; Pospisil, Heike; Kurtz, Stefan; Görndt, Jennifer; Püschel, Klaus; Riethdorf, Sabine; Pantel, Klaus; Brandt, Burkhard

2011-01-01

During cancer progression, specific genomic aberrations arise that can determine the scope of the disease and can be used as predictive or prognostic markers. The detection of specific gene amplifications or deletions in single blood-borne or disseminated tumour cells that may give rise to the development of metastases is of great clinical interest but technically challenging. In this study, we present a method for quantitative high-resolution genomic analysis of single cells. Cells were isolated under permanent microscopic control followed by high-fidelity whole genome amplification and subsequent analyses by fine tiling array-CGH and qPCR. The assay was applied to single breast cancer cells to analyze the chromosomal region centred by the therapeutical relevant EGFR gene. This method allows precise quantitative analysis of copy number variations in single cell diagnostics.

Quantitative Proteomic Analysis of Sulfolobus solfataricus Membrane Proteins

NARCIS (Netherlands)

Pham, T.K.; Sierocinski, P.; Oost, van der J.; Wright, P.C.

2010-01-01

A quantitative proteomic analysis of the membrane of the archaeon Sulfolobus solfataricus P2 using iTRAQ was successfully demonstrated in this technical note. The estimated number of membrane proteins of this organism is 883 (predicted based on Gravy score), corresponding to 30 % of the total

Quantitative X-ray analysis of biological fluids: the microdroplet technique

International Nuclear Information System (INIS)

Roinel, N.

1988-01-01

X-ray microanalysis can be used to quantitatively determine the elemental composition of microvolumes of biological fluids. This article describes the various steps in preparation of microdroplets for analysis: The manufacturing of micropipettes, the preparation of the specimen support, the deposition of droplets on the support, shock-freezing, and lyophilization. Examples of common artifacts (incomplete rehydration prior to freezing or partial rehydration after lyophilization) are demonstrated. Analysis can be carried out either by wavelength-dispersive analysis, which is the most sensitive method, or by energy-dispersive analysis, which is more commonly available. The minimum detectable concentration is 0.05 mmol.liter-1 for 0.1-nl samples analyzed by wavelength-dispersive spectrometry and 0.5-1 mmol.liter-1 for samples analyzed by energy-dispersive spectrometry. A major problem, especially in wavelength-dispersive analysis, where high beam currents are used, is radiation damage to the specimen; in particular chloride (but also other elements) can be lost. Quantitative analysis requires the use of standard solutions with elemental concentration in the same range as those present in the specimen

Method of quantitative x-ray diffractometric analysis of Ta-Ta2C system

International Nuclear Information System (INIS)

Gavrish, A.A.; Glazunov, M.P.; Korolev, Yu.M.; Spitsyn, V.I.; Fedoseev, G.K.

1976-01-01

The syste86 Ta-Ta 2 C has beemonsidered because of specific features of diffraction patterns of the components, namely, overlapping of the most intensive reflexes of both phases. The method of standard binary system has been used for quantitative analysis. Because of overlapping of the intensive reflexes dsub(1/01)=2.36(Ta 2 C) and dsub(110)=2.33(Ta), the other, most intensive, reflexes have been used for quantitative determination of Ta 2 C and Ta: dsub(103)=1.404 A for tantalum subcarbide and dsub(211)=1.35A for tantalum. Besides, the TaTa 2 C phases have been determined quantitatively with the use of another pair of reflexes: dsub(102)=1.82 A for Ta 2 C and dsub(200)=1.65 A for tantalum. The agreement between the results obtained while performing the quantitative phase analysis is good. To increase reliability and accuracy of the quantitative determination of Ta and Ta 2 C, it is expedient to carry out the analysis with the use of two above-mentioned pairs of reflexes located in different regions of the diffraction spectrum. Thus, the procedure of quantitative analysis of Ta and Ta 2 C in different ratios has been developed taking into account the specific features of the diffraction patterns of these components as well as the ability of Ta 2 C to texture in the process of preparation

A Quantitative Analysis of the Behavioral Checklist of the Movement ABC Motor Test

Science.gov (United States)

Ruiz, Luis Miguel; Gomez, Marta; Graupera, Jose Luis; Gutierrez, Melchor; Linaza, Jose Luis

2007-01-01

The fifth section of the Henderson and Sugden's Movement ABC Checklist is part of the general Checklist that accompanies The Movement ABC Battery. The authors maintain that the analysis of this section must be mainly qualitative instead of quantitative. The main objective of this study was to employ a quantitative analysis of this behavioural…

Quantitative assessment of in-solution digestion efficiency identifies optimal protocols for unbiased protein analysis

DEFF Research Database (Denmark)

Leon, Ileana R; Schwämmle, Veit; Jensen, Ole N

2013-01-01

a combination of qualitative and quantitative LC-MS/MS methods and statistical data analysis. In contrast to previous studies we employed both standard qualitative as well as data-independent quantitative workflows to systematically assess trypsin digestion efficiency and bias using mitochondrial protein...... conditions (buffer, RapiGest, deoxycholate, urea), and two methods for removal of detergents prior to analysis of peptides (acid precipitation or phase separation with ethyl acetate). Our data-independent quantitative LC-MS/MS workflow quantified over 3700 distinct peptides with 96% completeness between all...... protocols and replicates, with an average 40% protein sequence coverage and an average of 11 peptides identified per protein. Systematic quantitative and statistical analysis of physicochemical parameters demonstrated that deoxycholate-assisted in-solution digestion combined with phase transfer allows...

Chemical Fingerprint Analysis and Quantitative Analysis of Rosa rugosa by UPLC-DAD

Directory of Open Access Journals (Sweden)

Sanawar Mansur

2016-12-01

Full Text Available A method based on ultra performance liquid chromatography with a diode array detector (UPLC-DAD was developed for quantitative analysis of five active compounds and chemical fingerprint analysis of Rosa rugosa. Ten batches of R. rugosa collected from different plantations in the Xinjiang region of China were used to establish the fingerprint. The feasibility and advantages of the used UPLC fingerprint were verified for its similarity evaluation by systematically comparing chromatograms with professional analytical software recommended by State Food and Drug Administration (SFDA of China. In quantitative analysis, the five compounds showed good regression (R2 = 0.9995 within the test ranges, and the recovery of the method was in the range of 94.2%–103.8%. The similarities of liquid chromatography fingerprints of 10 batches of R. rugosa were more than 0.981. The developed UPLC fingerprint method is simple, reliable, and validated for the quality control and identification of R. rugosa. Additionally, simultaneous quantification of five major bioactive ingredients in the R. rugosa samples was conducted to interpret the consistency of the quality test. The results indicated that the UPLC fingerprint, as a characteristic distinguishing method combining similarity evaluation and quantification analysis, can be successfully used to assess the quality and to identify the authenticity of R. rugosa.

Phase analysis in duplex stainless steel: comparison of EBSD and quantitative metallography methods

International Nuclear Information System (INIS)

Michalska, J; Chmiela, B

2014-01-01

The purpose of the research was to work out the qualitative and quantitative analysis of phases in DSS in as-received state and after thermal aging. For quantitative purposes, SEM observations, EDS analyses and electron backscattered diffraction (EBSD) methods were employed. Qualitative analysis of phases was performed by two methods: EBSD and classical quantitative metallography. A juxtaposition of different etchants for the revealing of microstructure and brief review of sample preparation methods for EBSD studies were presented. Different ways of sample preparation were tested and based on these results a detailed methodology of DSS phase analysis was developed including: surface finishing, selective etching methods and image acquisition. The advantages and disadvantages of applied methods were pointed out and compared the accuracy of the analysis phase performed by both methods

Comparative study of standard space and real space analysis of quantitative MR brain data.

Science.gov (United States)

Aribisala, Benjamin S; He, Jiabao; Blamire, Andrew M

2011-06-01

To compare the robustness of region of interest (ROI) analysis of magnetic resonance imaging (MRI) brain data in real space with analysis in standard space and to test the hypothesis that standard space image analysis introduces more partial volume effect errors compared to analysis of the same dataset in real space. Twenty healthy adults with no history or evidence of neurological diseases were recruited; high-resolution T(1)-weighted, quantitative T(1), and B(0) field-map measurements were collected. Algorithms were implemented to perform analysis in real and standard space and used to apply a simple standard ROI template to quantitative T(1) datasets. Regional relaxation values and histograms for both gray and white matter tissues classes were then extracted and compared. Regional mean T(1) values for both gray and white matter were significantly lower using real space compared to standard space analysis. Additionally, regional T(1) histograms were more compact in real space, with smaller right-sided tails indicating lower partial volume errors compared to standard space analysis. Standard space analysis of quantitative MRI brain data introduces more partial volume effect errors biasing the analysis of quantitative data compared to analysis of the same dataset in real space. Copyright © 2011 Wiley-Liss, Inc.

Statistical shape analysis using 3D Poisson equation--A quantitatively validated approach.

Science.gov (United States)

Gao, Yi; Bouix, Sylvain

2016-05-01

Statistical shape analysis has been an important area of research with applications in biology, anatomy, neuroscience, agriculture, paleontology, etc. Unfortunately, the proposed methods are rarely quantitatively evaluated, and as shown in recent studies, when they are evaluated, significant discrepancies exist in their outputs. In this work, we concentrate on the problem of finding the consistent location of deformation between two population of shapes. We propose a new shape analysis algorithm along with a framework to perform a quantitative evaluation of its performance. Specifically, the algorithm constructs a Signed Poisson Map (SPoM) by solving two Poisson equations on the volumetric shapes of arbitrary topology, and statistical analysis is then carried out on the SPoMs. The method is quantitatively evaluated on synthetic shapes and applied on real shape data sets in brain structures. Copyright © 2016 Elsevier B.V. All rights reserved.

Validation of FRET Assay for the Screening of Growth Inhibitors of Escherichia coli Reveals Elongasome Assembly Dynamics

Science.gov (United States)

van der Ploeg, René; Goudelis, Spyridon Theodoros; den Blaauwen, Tanneke

2015-01-01

The increase in antibiotic resistant bacteria demands the development of new antibiotics against preferably new targets. The common approach is to test compounds for their ability to kill bacteria or to design molecules that inhibit essential protein activities in vitro. In the first case, the mode of action of the drug is unknown and in the second case, it is not known whether the compound will pass the impermeable barrier of the bacterial envelope. We developed an assay that detects the target of a compound, as well as its ability to pass the membrane(s) simultaneously. The Escherichia coli cytoskeletal protein MreB recruits protein complexes (elongasomes) that are essential for cell envelope growth. An in cell Förster Resonance Energy Transfer (FRET) assay was developed to detect the interaction between MreB molecules and between MreB and the elongasome proteins RodZ, RodA and PBP2. Inhibition of the polymerization of MreB by S-(3,4-dichlorobenzyl) isothiourea (A22) or of the activity of PBP2 by mecilinam resulted in loss or reduction of all measured interactions. This suggests that the interactions between the elongasome proteins are governed by a combination of weak affinities and substrate availability. This validated in cell FRET assay can be used to screen for cell envelope growth inhibitors. PMID:26263980

Validation of FRET Assay for the Screening of Growth Inhibitors of Escherichia coli Reveals Elongasome Assembly Dynamics

Directory of Open Access Journals (Sweden)

René van der Ploeg

2015-07-01

Full Text Available The increase in antibiotic resistant bacteria demands the development of new antibiotics against preferably new targets. The common approach is to test compounds for their ability to kill bacteria or to design molecules that inhibit essential protein activities in vitro. In the first case, the mode of action of the drug is unknown and in the second case, it is not known whether the compound will pass the impermeable barrier of the bacterial envelope. We developed an assay that detects the target of a compound, as well as its ability to pass the membrane(s simultaneously. The Escherichia coli cytoskeletal protein MreB recruits protein complexes (elongasomes that are essential for cell envelope growth. An in cell Förster Resonance Energy Transfer (FRET assay was developed to detect the interaction between MreB molecules and between MreB and the elongasome proteins RodZ, RodA and PBP2. Inhibition of the polymerization of MreB by S-(3,4-dichlorobenzyl isothiourea (A22 or of the activity of PBP2 by mecilinam resulted in loss or reduction of all measured interactions. This suggests that the interactions between the elongasome proteins are governed by a combination of weak affinities and substrate availability. This validated in cell FRET assay can be used to screen for cell envelope growth inhibitors.

Quantitative analysis of background parenchymal enhancement in whole breast on MRI: Influence of menstrual cycle and comparison with a qualitative analysis.

Science.gov (United States)

Jung, Yongsik; Jeong, Seong Kyun; Kang, Doo Kyoung; Moon, Yeorae; Kim, Tae Hee

2018-06-01

We quantitatively analyzed background parenchymal enhancement (BPE) in whole breast according to menstrual cycle and compared it with a qualitative analysis method. A data set of breast magnetic resonance imaging (MRI) from 273 breast cancer patients was used. For quantitative analysis, we used semiautomated in-house software with MATLAB. From each voxel of whole breast, the software calculated BPE using following equation: [(signal intensity [SI] at 1 min 30 s after contrast injection - baseline SI)/baseline SI] × 100%. In total, 53 patients had minimal, 108 mild, 87 moderate, and 25 marked BPE. On quantitative analysis, mean BPE values were 33.1% in the minimal, 42.1% in the mild, 59.1% in the moderate, and 81.9% in the marked BPE group showing significant difference (p = .009 for minimal vs. mild, p quantitative BPE (r = 0.63, p Quantitative analysis of BPE correlated well with the qualitative BPE grade. Quantitative BPE values were lowest in the second week and highest in the fourth week. Copyright © 2018 Elsevier B.V. All rights reserved.

Towards quantitative laser-induced breakdown spectroscopy analysis of soil samples

International Nuclear Information System (INIS)

Bousquet, B.; Sirven, J.-B.; Canioni, L.

2007-01-01

A quantitative analysis of chromium in soil samples is presented. Different emission lines related to chromium are studied in order to select the best one for quantitative features. Important matrix effects are demonstrated from one soil to the other, preventing any prediction of concentration in different soils on the basis of a univariate calibration curve. Finally, a classification of the LIBS data based on a series of Principal Component Analyses (PCA) is applied to a reduced dataset of selected spectral lines related to the major chemical elements in the soils. LIBS data of heterogeneous soils appear to be widely dispersed, which leads to a reconsideration of the sampling step in the analysis process

Quantitative high-resolution genomic analysis of single cancer cells.

Directory of Open Access Journals (Sweden)

Juliane Hannemann

Full Text Available During cancer progression, specific genomic aberrations arise that can determine the scope of the disease and can be used as predictive or prognostic markers. The detection of specific gene amplifications or deletions in single blood-borne or disseminated tumour cells that may give rise to the development of metastases is of great clinical interest but technically challenging. In this study, we present a method for quantitative high-resolution genomic analysis of single cells. Cells were isolated under permanent microscopic control followed by high-fidelity whole genome amplification and subsequent analyses by fine tiling array-CGH and qPCR. The assay was applied to single breast cancer cells to analyze the chromosomal region centred by the therapeutical relevant EGFR gene. This method allows precise quantitative analysis of copy number variations in single cell diagnostics.

Time-resolved UV-excited microarray reader for fluorescence energy transfer (FRET) measurements

Science.gov (United States)

Orellana, Adelina; Hokkanen, Ari P.; Pastinen, Tomi; Takkinen, Kristina; Soderlund, Hans

2001-05-01

Analytical systems based on immunochemistry are largely used in medical diagnostics and in biotechnology. There is a significant pressure to develop the present assay formats to become easier to use, faster, and less reagent consuming. Further developments towards high density array--like multianalyte measurement systems would be valuable. To this aim we have studied the applicability of fluorescence resonance energy transfer and time-resolved fluorescence resonance energy transfer in immunoassays on microspots and in microwells. We have used engineered recombinant antibodies detecting the pentameric protein CRP as a model analyte system, and tested different assay formats. We describe also the construction of a time-resolved scanning epifluorometer with which we could measure the FRET interaction between the slow fluorescence decay from europium chelates and its energy transfer to the rapidly decaying fluorophore Cy5.

Quantitative risk analysis as a basis for emergency planning

Energy Technology Data Exchange (ETDEWEB)

Yogui, Regiane Tiemi Teruya [Bureau Veritas do Brasil, Rio de Janeiro, RJ (Brazil); Macedo, Eduardo Soares de [Instituto de Pesquisas Tecnologicas (IPT), Sao Paulo, SP (Brazil)

2009-07-01

Several environmental accidents happened in Brazil and in the world during the 70's and 80's. This strongly motivated the preparation for emergencies in the chemical and petrochemical industries. Environmental accidents affect the environment and the communities that are neighbor to the industrial facilities. The present study aims at subsidizing and providing orientation to develop Emergency Planning from the data obtained on Quantitative Risk Analysis, elaborated according to the Technical Standard P4.261/03 from CETESB (Sao Paulo Environmental Agency). It was observed, during the development of the research, that the data generated on these studies need a complementation and a deeper analysis, so that it is possible to use them on the Emergency Plans. The main issues that were analyzed and discussed on this study were the reevaluation of hazard identification for the emergency plans, the consequences and vulnerability analysis for the response planning, the risk communication, and the preparation to respond to the emergencies of the communities exposed to manageable risks. As a result, the study intends to improve the interpretation and use of the data deriving from the Quantitative Risk Analysis to develop the emergency plans. (author)

A Strategy for Identifying Quantitative Trait Genes Using Gene Expression Analysis and Causal Analysis

Directory of Open Access Journals (Sweden)

Akira Ishikawa

2017-11-01

Full Text Available Large numbers of quantitative trait loci (QTL affecting complex diseases and other quantitative traits have been reported in humans and model animals. However, the genetic architecture of these traits remains elusive due to the difficulty in identifying causal quantitative trait genes (QTGs for common QTL with relatively small phenotypic effects. A traditional strategy based on techniques such as positional cloning does not always enable identification of a single candidate gene for a QTL of interest because it is difficult to narrow down a target genomic interval of the QTL to a very small interval harboring only one gene. A combination of gene expression analysis and statistical causal analysis can greatly reduce the number of candidate genes. This integrated approach provides causal evidence that one of the candidate genes is a putative QTG for the QTL. Using this approach, I have recently succeeded in identifying a single putative QTG for resistance to obesity in mice. Here, I outline the integration approach and discuss its usefulness using my studies as an example.

A Strategy for Identifying Quantitative Trait Genes Using Gene Expression Analysis and Causal Analysis.

Science.gov (United States)

Ishikawa, Akira

2017-11-27

Large numbers of quantitative trait loci (QTL) affecting complex diseases and other quantitative traits have been reported in humans and model animals. However, the genetic architecture of these traits remains elusive due to the difficulty in identifying causal quantitative trait genes (QTGs) for common QTL with relatively small phenotypic effects. A traditional strategy based on techniques such as positional cloning does not always enable identification of a single candidate gene for a QTL of interest because it is difficult to narrow down a target genomic interval of the QTL to a very small interval harboring only one gene. A combination of gene expression analysis and statistical causal analysis can greatly reduce the number of candidate genes. This integrated approach provides causal evidence that one of the candidate genes is a putative QTG for the QTL. Using this approach, I have recently succeeded in identifying a single putative QTG for resistance to obesity in mice. Here, I outline the integration approach and discuss its usefulness using my studies as an example.

Quantitative analysis of patient-specific dosimetric IMRT verification

International Nuclear Information System (INIS)

Budgell, G J; Perrin, B A; Mott, J H L; Fairfoul, J; Mackay, R I

2005-01-01

Patient-specific dosimetric verification methods for IMRT treatments are variable, time-consuming and frequently qualitative, preventing evidence-based reduction in the amount of verification performed. This paper addresses some of these issues by applying a quantitative analysis parameter to the dosimetric verification procedure. Film measurements in different planes were acquired for a series of ten IMRT prostate patients, analysed using the quantitative parameter, and compared to determine the most suitable verification plane. Film and ion chamber verification results for 61 patients were analysed to determine long-term accuracy, reproducibility and stability of the planning and delivery system. The reproducibility of the measurement and analysis system was also studied. The results show that verification results are strongly dependent on the plane chosen, with the coronal plane particularly insensitive to delivery error. Unexpectedly, no correlation could be found between the levels of error in different verification planes. Longer term verification results showed consistent patterns which suggest that the amount of patient-specific verification can be safely reduced, provided proper caution is exercised: an evidence-based model for such reduction is proposed. It is concluded that dose/distance to agreement (e.g., 3%/3 mm) should be used as a criterion of acceptability. Quantitative parameters calculated for a given criterion of acceptability should be adopted in conjunction with displays that show where discrepancies occur. Planning and delivery systems which cannot meet the required standards of accuracy, reproducibility and stability to reduce verification will not be accepted by the radiotherapy community

Fluorescent foci quantitation for high-throughput analysis

Directory of Open Access Journals (Sweden)

Elena Ledesma-Fernández

2015-06-01

Full Text Available A number of cellular proteins localize to discrete foci within cells, for example DNA repair proteins, microtubule organizing centers, P bodies or kinetochores. It is often possible to measure the fluorescence emission from tagged proteins within these foci as a surrogate for the concentration of that specific protein. We wished to develop tools that would allow quantitation of fluorescence foci intensities in high-throughput studies. As proof of principle we have examined the kinetochore, a large multi-subunit complex that is critical for the accurate segregation of chromosomes during cell division. Kinetochore perturbations lead to aneuploidy, which is a hallmark of cancer cells. Hence, understanding kinetochore homeostasis and regulation are important for a global understanding of cell division and genome integrity. The 16 budding yeast kinetochores colocalize within the nucleus to form a single focus. Here we have created a set of freely-available tools to allow high-throughput quantitation of kinetochore foci fluorescence. We use this ‘FociQuant’ tool to compare methods of kinetochore quantitation and we show proof of principle that FociQuant can be used to identify changes in kinetochore protein levels in a mutant that affects kinetochore function. This analysis can be applied to any protein that forms discrete foci in cells.

A simple approach to quantitative analysis using three-dimensional spectra based on selected Zernike moments.

Science.gov (United States)

Zhai, Hong Lin; Zhai, Yue Yuan; Li, Pei Zhen; Tian, Yue Li

2013-01-21

A very simple approach to quantitative analysis is proposed based on the technology of digital image processing using three-dimensional (3D) spectra obtained by high-performance liquid chromatography coupled with a diode array detector (HPLC-DAD). As the region-based shape features of a grayscale image, Zernike moments with inherently invariance property were employed to establish the linear quantitative models. This approach was applied to the quantitative analysis of three compounds in mixed samples using 3D HPLC-DAD spectra, and three linear models were obtained, respectively. The correlation coefficients (R(2)) for training and test sets were more than 0.999, and the statistical parameters and strict validation supported the reliability of established models. The analytical results suggest that the Zernike moment selected by stepwise regression can be used in the quantitative analysis of target compounds. Our study provides a new idea for quantitative analysis using 3D spectra, which can be extended to the analysis of other 3D spectra obtained by different methods or instruments.

Single particle transfer for quantitative analysis with total-reflection X-ray fluorescence spectrometry

International Nuclear Information System (INIS)

Esaka, Fumitaka; Esaka, Konomi T.; Magara, Masaaki; Sakurai, Satoshi; Usuda, Shigekazu; Watanabe, Kazuo

2006-01-01

The technique of single particle transfer was applied to quantitative analysis with total-reflection X-ray fluorescence (TXRF) spectrometry. The technique was evaluated by performing quantitative analysis of individual Cu particles with diameters between 3.9 and 13.2 μm. The direct quantitative analysis of the Cu particle transferred onto a Si carrier gave a discrepancy between measured and calculated Cu amounts due to the absorption effects of incident and fluorescent X-rays within the particle. By the correction for the absorption effects, the Cu amounts in individual particles could be determined with the deviation within 10.5%. When the Cu particles were dissolved with HNO 3 solution prior to the TXRF analysis, the deviation was improved to be within 3.8%. In this case, no correction for the absorption effects was needed for quantification

Improving the spectral analysis of Fluorescence Resonance Energy Transfer in live cells: application to interferon receptors and Janus kinases.

Science.gov (United States)

Krause, Christopher D; Digioia, Gina; Izotova, Lara S; Pestka, Sidney

2013-10-01

The observed Fluorescence Resonance Energy Transfer (FRET) between fluorescently labeled proteins varies in cells. To understand how this variation affects our interpretation of how proteins interact in cells, we developed a protocol that mathematically separates donor-independent and donor-dependent excitations of acceptor, determines the electromagnetic interaction of donors and acceptors, and quantifies the efficiency of the interaction of donors and acceptors. By analyzing large populations of cells, we found that misbalanced or insufficient expression of acceptor or donor as well as their inefficient or reversible interaction influenced FRET efficiency in vivo. Use of red-shifted donors and acceptors gave spectra with less endogenous fluorescence but produced lower FRET efficiency, possibly caused by reduced quenching of red-shifted fluorophores in cells. Additionally, cryptic interactions between jellyfish FPs artefactually increased the apparent FRET efficiency. Our protocol can distinguish specific and nonspecific protein interactions even within highly constrained environments as plasma membranes. Overall, accurate FRET estimations in cells or within complex environments can be obtained by a combination of proper data analysis, study of sufficient numbers of cells, and use of properly empirically developed fluorescent proteins. Copyright © 2013 Elsevier Ltd. All rights reserved.

Quantitative analysis of thallium-201 myocardial scintigraphy

International Nuclear Information System (INIS)

Kanemoto, Nariaki; Hoer, G.; Johost, S.; Maul, F.-D.; Standke, R.

1981-01-01

The method of quantitative analysis of thallium-201 myocardial scintigraphy using computer assisted technique was described. Calculated indices are washout factor, vitality index and redistribution factor. Washout factor is the ratio of counts at certain period of time after exercise and immediately after exercise. This value is neccessary for the evaluation of redistribution to the ischemic areas in serial imagings to correct the Tl-201 washout from the myocardium under the assumption that the washout is constant in the whole myocardium. Vitality index is the ratio between the Tl-201 uptake in the region of interest and that of the maximum. Redistribution factor is the ratio of the redistribution in the region of interest in serial imagings after exercise to that of immediately after exercise. Four examples of exercise Tl-201 myocardial scintigrams and the quantitative analyses before and after the percutaneous transluminal coronary angioplasty were presented. (author)

Vibration analysis and vibration damage assessment in nuclear and process equipment

International Nuclear Information System (INIS)

Pettigrew, M.J.; Taylor, C.E.; Fisher, N.J.; Yetisir, M.; Smith, B.A.W.

1997-01-01

Component failures due to excessive flow-induced vibration are still affecting the performance and reliability of process and nuclear components. The purpose of this paper is to discuss flow-induced vibration analysis and vibration damage prediction. Vibration excitation mechanisms are described with particular emphasis on fluid elastic instability. The dynamic characteristics of process and power equipment are explained. The statistical nature of some parameters, in particular support conditions, is discussed. The prediction of fretting-wear damage is approached from several points-of-view. An energy approach to formulate fretting-wear damage is proposed. (author)

Quantitative pulsed eddy current analysis

International Nuclear Information System (INIS)

Morris, R.A.

1975-01-01

The potential of pulsed eddy current testing for furnishing more information than conventional single-frequency eddy current methods has been known for some time. However, a fundamental problem has been analyzing the pulse shape with sufficient precision to produce accurate quantitative results. Accordingly, the primary goal of this investigation was to: demonstrate ways of digitizing the short pulses encountered in PEC testing, and to develop empirical analysis techniques that would predict some of the parameters (e.g., depth) of simple types of defect. This report describes a digitizing technique using a computer and either a conventional nuclear ADC or a fast transient analyzer; the computer software used to collect and analyze pulses; and some of the results obtained. (U.S.)

Quantitative surface analysis using deuteron-induced nuclear reactions

International Nuclear Information System (INIS)

Afarideh, Hossein

1991-01-01

The nuclear reaction analysis (NRA) technique consists of looking at the energies of the reaction products which uniquely define the particular elements present in the sample and it analysis the yield/energy distribution to reveal depth profiles. A summary of the basic features of the nuclear reaction analysis technique is given, in particular emphasis is placed on quantitative light element determination using (d,p) and (d,alpha) reactions. The experimental apparatus is also described. Finally a set of (d,p) spectra for the elements Z=3 to Z=17 using 2 MeV incident deutrons is included together with example of more applications of the (d,alpha) spectra. (author)

Evaluation of shear wave elastography for differential diagnosis of breast lesions: A new qualitative analysis versus conventional quantitative analysis.

Science.gov (United States)

Ren, Wei-Wei; Li, Xiao-Long; Wang, Dan; Liu, Bo-Ji; Zhao, Chong-Ke; Xu, Hui-Xiong

2018-04-13

To evaluate a special kind of ultrasound (US) shear wave elastography for differential diagnosis of breast lesions, using a new qualitative analysis (i.e. the elasticity score in the travel time map) compared with conventional quantitative analysis. From June 2014 to July 2015, 266 pathologically proven breast lesions were enrolled in this study. The maximum, mean, median, minimum, and standard deviation of shear wave speed (SWS) values (m/s) were assessed. The elasticity score, a new qualitative feature, was evaluated in the travel time map. The area under the receiver operating characteristic (AUROC) curves were plotted to evaluate the diagnostic performance of both qualitative and quantitative analyses for differentiation of breast lesions. Among all quantitative parameters, SWS-max showed the highest AUROC (0.805; 95% CI: 0.752, 0.851) compared with SWS-mean (0.786; 95% CI:0.732, 0.834; P = 0.094), SWS-median (0.775; 95% CI:0.720, 0.824; P = 0.046), SWS-min (0.675; 95% CI:0.615, 0.731; P = 0.000), and SWS-SD (0.768; 95% CI:0.712, 0.817; P = 0.074). The AUROC of qualitative analysis in this study obtained the best diagnostic performance (0.871; 95% CI: 0.825, 0.909, compared with the best parameter of SWS-max in quantitative analysis, P = 0.011). The new qualitative analysis of shear wave travel time showed the superior diagnostic performance in the differentiation of breast lesions in comparison with conventional quantitative analysis.

MCM - 2 and Ki - 67 as proliferation markers in renal cell carcinoma: A quantitative and semi - quantitative analysis.

Science.gov (United States)

Mehdi, Muhammad Zain; Nagi, Abdul Hanan; Naseem, Nadia

2016-01-01

Fuhrman nuclear grade is the most important histological parameter to predict prognosis in a patient of renal cell carcinoma (RCC). However, it suffers from inter-observer and intra-observer variation giving rise to need of a parameter that not only correlates with nuclear grade but is also objective and reproducible. Proliferation is the measure of aggressiveness of a tumour and it is strongly correlated with Fuhrman nuclear grade, clinical survival and recurrence in RCC. Ki-67 is conventionally used to assess proliferation. Mini-chromosome maintenance 2 (MCM-2) is a lesser known marker of proliferation and identifies a greater proliferation faction. This study was designed to assess the prognostic significance of MCM-2 by comparing it with Fuhrman nuclear grade and Ki-67. n=50 cases of various ages, stages, histological subtypes and grades of RCC were selected for this study. Immunohistochemical staining using Ki-67(MIB-1, Mouse monoclonal antibody, Dako) and MCM-2 (Mouse monoclonal antibody, Thermo) was performed on the paraffin embedded blocks in the department of Morbid anatomy and Histopathology, University of Health Sciences, Lahore. Labeling indices (LI) were determined by two pathologists independently using quantitative and semi-quantitative analysis. Statistical analysis was carried out using SPSS 20.0. Kruskall-Wallis test was used to determine a correlation of proliferation markers with grade, and Pearson's correlate was used to determine correlation between the two proliferation markers. Labeling index of MCM-2 (median=24.29%) was found to be much higher than Ki-67(median=13.05%). Both markers were significantly related with grade (p=0.00; Kruskall-Wallis test). LI of MCM-2 was found to correlate significantly with LI of Ki-67(r=0.0934;p=0.01 with Pearson's correlate). Results of semi-quantitative analysis correlated well with quantitative analysis. Both Ki-67 and MCM-2 are markers of proliferation which are closely linked to grade. Therefore, they

Quantitative Myocardial Perfusion Imaging Versus Visual Analysis in Diagnosing Myocardial Ischemia: A CE-MARC Substudy.

Science.gov (United States)

Biglands, John D; Ibraheem, Montasir; Magee, Derek R; Radjenovic, Aleksandra; Plein, Sven; Greenwood, John P

2018-05-01

This study sought to compare the diagnostic accuracy of visual and quantitative analyses of myocardial perfusion cardiovascular magnetic resonance against a reference standard of quantitative coronary angiography. Visual analysis of perfusion cardiovascular magnetic resonance studies for assessing myocardial perfusion has been shown to have high diagnostic accuracy for coronary artery disease. However, only a few small studies have assessed the diagnostic accuracy of quantitative myocardial perfusion. This retrospective study included 128 patients randomly selected from the CE-MARC (Clinical Evaluation of Magnetic Resonance Imaging in Coronary Heart Disease) study population such that the distribution of risk factors and disease status was proportionate to the full population. Visual analysis results of cardiovascular magnetic resonance perfusion images, by consensus of 2 expert readers, were taken from the original study reports. Quantitative myocardial blood flow estimates were obtained using Fermi-constrained deconvolution. The reference standard for myocardial ischemia was a quantitative coronary x-ray angiogram stenosis severity of ≥70% diameter in any coronary artery of >2 mm diameter, or ≥50% in the left main stem. Diagnostic performance was calculated using receiver-operating characteristic curve analysis. The area under the curve for visual analysis was 0.88 (95% confidence interval: 0.81 to 0.95) with a sensitivity of 81.0% (95% confidence interval: 69.1% to 92.8%) and specificity of 86.0% (95% confidence interval: 78.7% to 93.4%). For quantitative stress myocardial blood flow the area under the curve was 0.89 (95% confidence interval: 0.83 to 0.96) with a sensitivity of 87.5% (95% confidence interval: 77.3% to 97.7%) and specificity of 84.5% (95% confidence interval: 76.8% to 92.3%). There was no statistically significant difference between the diagnostic performance of quantitative and visual analyses (p = 0.72). Incorporating rest myocardial

A critical appraisal of semi-quantitative analysis of 2-deoxyglucose autoradiograms

International Nuclear Information System (INIS)

Kelly, P.T.; McCulloch, J.

1983-01-01

Semi-quantitative analysis (e.g. optical density ratios) of [ 14 C]2-deoxyglucose autoradiograms is widely used in neuroscience research. The authors demonstrate that a fixed ratio of 14 C-concentrations in the CNS does not yield a constant optical density ratio but is dependent upon the exposure time in the preparation of the autoradiograms and the absolute amounts of 14 C from which the concentration ratio is derived. The failure of a fixed glucose utilization ratio to result in a constant optical density ratio represents a major interpretative difficulty in investigations where only semi-quantitative analysis of [ 14 C]2-deoxyglucose autoradiograms is undertaken. (Auth.)

Quantitative analysis of culture using millions of digitized books.

Science.gov (United States)

Michel, Jean-Baptiste; Shen, Yuan Kui; Aiden, Aviva Presser; Veres, Adrian; Gray, Matthew K; Pickett, Joseph P; Hoiberg, Dale; Clancy, Dan; Norvig, Peter; Orwant, Jon; Pinker, Steven; Nowak, Martin A; Aiden, Erez Lieberman

2011-01-14

We constructed a corpus of digitized texts containing about 4% of all books ever printed. Analysis of this corpus enables us to investigate cultural trends quantitatively. We survey the vast terrain of 'culturomics,' focusing on linguistic and cultural phenomena that were reflected in the English language between 1800 and 2000. We show how this approach can provide insights about fields as diverse as lexicography, the evolution of grammar, collective memory, the adoption of technology, the pursuit of fame, censorship, and historical epidemiology. Culturomics extends the boundaries of rigorous quantitative inquiry to a wide array of new phenomena spanning the social sciences and the humanities.

Quantitative analysis of culture using millions of digitized books

Science.gov (United States)

Michel, Jean-Baptiste; Shen, Yuan Kui; Aiden, Aviva P.; Veres, Adrian; Gray, Matthew K.; Pickett, Joseph P.; Hoiberg, Dale; Clancy, Dan; Norvig, Peter; Orwant, Jon; Pinker, Steven; Nowak, Martin A.; Aiden, Erez Lieberman

2011-01-01

We constructed a corpus of digitized texts containing about 4% of all books ever printed. Analysis of this corpus enables us to investigate cultural trends quantitatively. We survey the vast terrain of ‘culturomics’, focusing on linguistic and cultural phenomena that were reflected in the English language between 1800 and 2000. We show how this approach can provide insights about fields as diverse as lexicography, the evolution of grammar, collective memory, the adoption of technology, the pursuit of fame, censorship, and historical epidemiology. ‘Culturomics’ extends the boundaries of rigorous quantitative inquiry to a wide array of new phenomena spanning the social sciences and the humanities. PMID:21163965

Quantitative mass spectrometric analysis of glycoproteins combined with enrichment methods.

Science.gov (United States)

Ahn, Yeong Hee; Kim, Jin Young; Yoo, Jong Shin

2015-01-01

Mass spectrometry (MS) has been a core technology for high sensitive and high-throughput analysis of the enriched glycoproteome in aspects of quantitative assays as well as qualitative profiling of glycoproteins. Because it has been widely recognized that aberrant glycosylation in a glycoprotein may involve in progression of a certain disease, the development of efficient analysis tool for the aberrant glycoproteins is very important for deep understanding about pathological function of the glycoprotein and new biomarker development. This review first describes the protein glycosylation-targeting enrichment technologies mainly employing solid-phase extraction methods such as hydrizide-capturing, lectin-specific capturing, and affinity separation techniques based on porous graphitized carbon, hydrophilic interaction chromatography, or immobilized boronic acid. Second, MS-based quantitative analysis strategies coupled with the protein glycosylation-targeting enrichment technologies, by using a label-free MS, stable isotope-labeling, or targeted multiple reaction monitoring (MRM) MS, are summarized with recent published studies. © 2014 The Authors. Mass Spectrometry Reviews Published by Wiley Periodicals, Inc.

Quantitative analysis of macro-ARG using IP system

International Nuclear Information System (INIS)

Nakajima, Eiichi; Kawai, Kenji; Furuta, Yoshitake

1997-01-01

Recent progress in imaging plate (IP) system allow us to analyze autoradiographic images quantitatively. In the 'whole-body autoradiography', a method clarify the distribution of radioisotope or labeled compounds in the tissues and organs in a freeze-dried whole-body section of small animals such as rats and mice, the sections are pressed against a IP for exposure and the IP is scanned by Bio-Imaging Analyzer (Fuji Photo Film Co., Ltd) and a digital autoradiographic image is given. Quantitative data concerning the activity in different tissues can be obtained using an isotope scale as a reference source. Fading effect, application of IP system for distribution of receptor binding ARG, analysis of radio-spots on TLC and radioactive concentration in liquid such as blood are also discussed. (author)

Potential Application of Quantitative Prostate-specific Antigen Analysis in Forensic Examination of Seminal Stains

Directory of Open Access Journals (Sweden)

Zhenping Liu

2015-01-01

Full Text Available The aims of this study are to use quantitative analysis of the prostate-specific antigen (PSA in the seminal stain examination and to explore the practical value of this analysis in forensic science. For a comprehensive analysis, vaginal swabs from 48 rape cases were tested both by a PSA fluorescence analyzer (i-CHROMA Reader and by a conventional PSA strip test. To confirm the results of these PSA tests, seminal DNA was tested following differential extraction. Compared to the PSA strip test, the PSA rapid quantitative fluorescence analyzer provided the more accurate and sensitive results. More importantly, individualized schemes based on quantitative PSA results of samples can be developed to improve the quality and procedural efficiency in the forensic seminal inspection of samples prior to DNA analysis.

On the Need for Quantitative Bias Analysis in the Peer-Review Process.

Science.gov (United States)

Fox, Matthew P; Lash, Timothy L

2017-05-15

Peer review is central to the process through which epidemiologists generate evidence to inform public health and medical interventions. Reviewers thereby act as critical gatekeepers to high-quality research. They are asked to carefully consider the validity of the proposed work or research findings by paying careful attention to the methodology and critiquing the importance of the insight gained. However, although many have noted problems with the peer-review system for both manuscripts and grant submissions, few solutions have been proposed to improve the process. Quantitative bias analysis encompasses all methods used to quantify the impact of systematic error on estimates of effect in epidemiologic research. Reviewers who insist that quantitative bias analysis be incorporated into the design, conduct, presentation, and interpretation of epidemiologic research could substantially strengthen the process. In the present commentary, we demonstrate how quantitative bias analysis can be used by investigators and authors, reviewers, funding agencies, and editors. By utilizing quantitative bias analysis in the peer-review process, editors can potentially avoid unnecessary rejections, identify key areas for improvement, and improve discussion sections by shifting from speculation on the impact of sources of error to quantification of the impact those sources of bias may have had. © The Author 2017. Published by Oxford University Press on behalf of the Johns Hopkins Bloomberg School of Public Health. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Contact damage failure analyses of fretting wear behavior of the metal stem titanium alloy-bone cement interface.

Science.gov (United States)

Zhang, Lanfeng; Ge, Shirong; Liu, Hongtao; Wang, Qingliang; Wang, Liping; Xian, Cory J

2015-11-01

Although cemented titanium alloy is not favored currently in the Western world for its poor clinical and radiography outcomes, its lower modulus of elasticity and good biocompatibility are instrumental for its ability supporting and transforming physical load, and it is more suitable for usage in Chinese and Japanese populations due to their lower body weights and unique femoral characteristics. Through various friction tests of different cycles, loads and conditions and by examining fretting hysteresis loops, fatigue process curves and wear surfaces, the current study investigated fretting wear characteristics and wear mechanism of titanium alloy stem-bone cement interface. It was found that the combination of loads and displacement affected the wear quantity. Friction coefficient, which was in an inverse relationship to load under the same amplitude, was proportional to amplitudes under the same load. Additionally, calf serum was found to both lubricate and erode the wear interface. Moreover, cement fatigue contact areas appeared black/oxidative in dry and gruel in 25% calf serum. Fatigue scratches were detected within contact areas, and wear scars were found on cement and titanium surfaces, which were concave-shaped and ring concave/ convex-shaped, respectively. The coupling of thermoplastic effect and minimal torque damage has been proposed to be the major reason of contact damage. These data will be important for further studies analyzing metal-cement interface failure performance and solving interface friction and wear debris production issues. Copyright © 2015 Elsevier Ltd. All rights reserved.

Immune adherence: a quantitative and kinetic analysis

Energy Technology Data Exchange (ETDEWEB)

Sekine, T [National Cancer Center, Tokyo (Japan). Research Inst.

1978-09-01

Quantitative and kinetic analysis of the immune-adherence reaction (IA) between C3b fragments and IA receptors as an agglutination reaction is difficult. Analysis is possible, however, by use of radio-iodinated bovine serum albumin as antigen at low concentrations (less than 200 ng/ml) and optimal concentration of antibody to avoid precipitation of antigen-antibody complexes with human erythrocytes without participation of complement. Antigen and antibody are reacted at 37/sup 0/C, complement is added, the mixture incubated and human erythrocytes added; after further incubation, ice-cold EDTA containing buffer is added and the erythrocytes centrifuged and assayed for radioactivity. Control cells reacted with heated guinea pig serum retained less than 5% of the added radioactivity. The method facilitates measurement of IA reactivity and permits more detailed analysis of the mechanism underlying the reaction.

Quantitative Determination of Aluminum in Deodorant Brands: A Guided Inquiry Learning Experience in Quantitative Analysis Laboratory

Science.gov (United States)

Sedwick, Victoria; Leal, Anne; Turner, Dea; Kanu, A. Bakarr

2018-01-01

The monitoring of metals in commercial products is essential for protecting public health against the hazards of metal toxicity. This article presents a guided inquiry (GI) experimental lab approach in a quantitative analysis lab class that enabled students' to determine the levels of aluminum in deodorant brands. The utility of a GI experimental…

Contribution to the study of atmospheric projection and under partial vacuum of tungsten carbide particles with cobalt or nickel binder. Application to fretting coatings on steel

International Nuclear Information System (INIS)

Vinayo, Maria-Elena

1985-01-01

This research thesis addresses the plasma spraying (atmospheric, under controlled atmosphere, and under reduced pressure) of tungsten carbides with a metallic binder (WC/Co, WC/Ni; W 2 C/Co). This work comprised an optimisation of the spraying process under reduced pressure, the study of the influence of the powder production process on the physicochemical and micro-structural characteristics as well as on coating fretting properties, and a correlation between spraying parameters in a controlled atmosphere (power and pressure) and coating physico-chemical and micro-structural properties. Results show a high decarburization-oxidation of tungsten carbides during atmospheric spraying, as well as an important evaporation of cobalt. Under reduced pressure, high losses of carbides are noticed. These both phenomena strongly depend on the powder production process. Fretting results highlight remarkable performance of coatings obtained by atmospheric spraying [fr

Quantitative data analysis with SPSS release 8 for Windows a guide for social scientists

CERN Document Server

Bryman, Alan

2002-01-01

The latest edition of this best-selling introduction to Quantitative Data Analysis through the use of a computer package has been completely updated to accommodate the needs of users of SPSS Release 8 for Windows. Like its predecessor, it provides a non-technical approach to quantitative data analysis and a user-friendly introduction to the widely used SPSS for Windows. It assumes no previous familiarity with either statistics or computing but takes the reader step-by-step through the techniques, reinforced by exercises for further practice. Techniques explained in Quantitative Data Analysis with SPSS Release 8 for Windows include: * correlation * simple and multiple regression * multivariate analysis of variance and covariance * factor analysis The book also covers issues such as sampling, statistical significance, conceptualization and measurement and the selection of appropriate tests. For further information or to download the book's datasets, please visit the webstite: http://www.routledge.com/textbooks/...

Quantitative analysis on electrooculography (EOG) for neurodegenerative disease

Science.gov (United States)

Liu, Chang-Chia; Chaovalitwongse, W. Art; Pardalos, Panos M.; Seref, Onur; Xanthopoulos, Petros; Sackellares, J. C.; Skidmore, Frank M.

2007-11-01

Many studies have documented abnormal horizontal and vertical eye movements in human neurodegenerative disease as well as during altered states of consciousness (including drowsiness and intoxication) in healthy adults. Eye movement measurement may play an important role measuring the progress of neurodegenerative diseases and state of alertness in healthy individuals. There are several techniques for measuring eye movement, Infrared detection technique (IR). Video-oculography (VOG), Scleral eye coil and EOG. Among those available recording techniques, EOG is a major source for monitoring the abnormal eye movement. In this real-time quantitative analysis study, the methods which can capture the characteristic of the eye movement were proposed to accurately categorize the state of neurodegenerative subjects. The EOG recordings were taken while 5 tested subjects were watching a short (>120 s) animation clip. In response to the animated clip the participants executed a number of eye movements, including vertical smooth pursued (SVP), horizontal smooth pursued (HVP) and random saccades (RS). Detection of abnormalities in ocular movement may improve our diagnosis and understanding a neurodegenerative disease and altered states of consciousness. A standard real-time quantitative analysis will improve detection and provide a better understanding of pathology in these disorders.

Quantitative XPS analysis of high Tc superconductor surfaces

International Nuclear Information System (INIS)

Jablonski, A.; Sanada, N.; Suzuki, Y.; Fukuda, Y.; Nagoshi, M.

1993-01-01

The procedure of quantitative XPS analysis involving the relative sensitivity factors is most convenient to apply to high T c superconductor surfaces because this procedure does not require standards. However, a considerable limitation of such an approach is its relatively low accuracy. In the present work, a proposition is made to use for this purpose a modification of the relative sensitivity factor approach accounting for the matrix and the instrumental effects. The accuracy of this modification when applied to the binary metal alloys is 2% or better. A quantitative XPS analysis was made for surfaces of the compounds Bi 2 Sr 2 CuO 6 , Bi 2 Sr 2 CaCu 2 O 8 , and YBa 2 Cu 3 O Y . The surface composition determined for the polycrystalline samples corresponds reasonably well to the bulk stoichiometry. Slight deficiency of oxygen was found for the Bi-based compounds. The surface exposed on cleavage of the Bi 2 Sr 2 CaCu 2 O 8 single crystal was found to be enriched with bismuth, which indicates that the cleavage occurs along the BiO planes. This result is in agreement with the STM studies published in the literature

Quantitative Analysis of Complex Tropical Forest Stands: A Review ...

African Journals Online (AJOL)

The importance of data analysis in quantitative assessment of natural resources remains significant in the sustainable management of complex tropical forest resources. Analyses of data from complex tropical forest stands have not been easy or clear due to improper data management. It is pivotal to practical researches ...

Fretting wear behaviour of TiC/Ti(C,N)/TiN multi-layer coatings at elevated temperature in gross slip regime

International Nuclear Information System (INIS)

Liu Hanwei; Huang Kunpeng; Zhu Minhao; Zhou Zhongrong

2005-01-01

Tic/Ti(C,N)/TiN multi-layer coatings are prepared on the 1Cr13 stainless steel substrate by the technique of Chemical Vapour Deposition, and the fretting wear behaviour of 1Cr13 stainless steel and TiC/Ti(C,N)/TiN coatings are investigated and studied controversially from 25 degree C to 400 degree C in the gross slip regime. It shows that the temperature has great influence on the fretting wear in the gross slip regime for the 1Cr13 stainless steel but little for Ti/C/Ti(C,N)/TiN multi-layer coatings. With the temperature increasing, the friction coefficient and the wear volume of the 1Cr13 alloy decreases and the wear volume of TiC/Ti(C, N)/TiN multi-layer coatings is invariant. TiC/Ti(C,N)/TiN multi-layer coatings have better wear-resistant capability than the 1Cr13 stainless steel, but the wear volume of the substrate increases greatly because of the grain-abrasion resulted from hard debris when TiC/Ti(C,N)/TiN multi-layer coatings are ground off. (authors)

Quantitative analysis of eyes and other optical systems in linear optics.

Science.gov (United States)

Harris, William F; Evans, Tanya; van Gool, Radboud D

2017-05-01

To show that 14-dimensional spaces of augmented point P and angle Q characteristics, matrices obtained from the ray transference, are suitable for quantitative analysis although only the latter define an inner-product space and only on it can one define distances and angles. The paper examines the nature of the spaces and their relationships to other spaces including symmetric dioptric power space. The paper makes use of linear optics, a three-dimensional generalization of Gaussian optics. Symmetric 2 × 2 dioptric power matrices F define a three-dimensional inner-product space which provides a sound basis for quantitative analysis (calculation of changes, arithmetic means, etc.) of refractive errors and thin systems. For general systems the optical character is defined by the dimensionally-heterogeneous 4 × 4 symplectic matrix S, the transference, or if explicit allowance is made for heterocentricity, the 5 × 5 augmented symplectic matrix T. Ordinary quantitative analysis cannot be performed on them because matrices of neither of these types constitute vector spaces. Suitable transformations have been proposed but because the transforms are dimensionally heterogeneous the spaces are not naturally inner-product spaces. The paper obtains 14-dimensional spaces of augmented point P and angle Q characteristics. The 14-dimensional space defined by the augmented angle characteristics Q is dimensionally homogenous and an inner-product space. A 10-dimensional subspace of the space of augmented point characteristics P is also an inner-product space. The spaces are suitable for quantitative analysis of the optical character of eyes and many other systems. Distances and angles can be defined in the inner-product spaces. The optical systems may have multiple separated astigmatic and decentred refracting elements. © 2017 The Authors Ophthalmic & Physiological Optics © 2017 The College of Optometrists.

Quantitative X ray analysis system. User's manual and guide to X ray fluorescence technique

International Nuclear Information System (INIS)

2009-01-01

This guide covers trimmed and re-arranged version 3.6 of the Quantitative X ray Analysis System (QXAS) software package that includes the most frequently used methods of quantitative analysis. QXAS is a comprehensive quantitative analysis package that has been developed by the IAEA through research and technical contracts. Additional development has also been carried out in the IAEA Laboratories in Seibersdorf where QXAS was extensively tested. New in this version of the manual are the descriptions of the Voigt-profile peak fitting, the backscatter fundamental parameters' and emission-transmission methods of chemical composition analysis, an expanded chapter on the X ray fluorescence physics, and completely revised and increased number of practical examples of utilization of the QXAS software package. The analytical data accompanying this manual were collected in the IAEA Seibersdorf Laboratories in the years 2006/2007

Quantitative proteomic analysis of intact plastids.

Science.gov (United States)

Shiraya, Takeshi; Kaneko, Kentaro; Mitsui, Toshiaki

2014-01-01

Plastids are specialized cell organelles in plant cells that are differentiated into various forms including chloroplasts, chromoplasts, and amyloplasts, and fulfill important functions in maintaining the overall cell metabolism and sensing environmental factors such as sunlight. It is therefore important to grasp the mechanisms of differentiation and functional changes of plastids in order to enhance the understanding of vegetality. In this chapter, details of a method for the extraction of intact plastids that makes analysis possible while maintaining the plastid functions are provided; in addition, a quantitative shotgun method for analyzing the composition and changes in the content of proteins in plastids as a result of environmental impacts is described.

Quantitative analysis of elastography images in the detection of breast cancer

International Nuclear Information System (INIS)

Landoni, V.; Francione, V.; Marzi, S.; Pasciuti, K.; Ferrante, F.; Saracca, E.; Pedrini, M.; Strigari, L.; Crecco, M.; Di Nallo, A.

2012-01-01

Purpose: The aim of this study was to develop a quantitative method for breast cancer diagnosis based on elastosonography images in order to reduce whenever possible unnecessary biopsies. The proposed method was validated by correlating the results of quantitative analysis with the diagnosis assessed by histopathologic exam. Material and methods: 109 images of breast lesions (50 benign and 59 malignant) were acquired with the traditional B-mode technique and with elastographic modality. Images in Digital Imaging and COmmunications in Medicine format (DICOM) were exported into a software, written in Visual Basic, especially developed to perform this study. The lesion was contoured and the mean grey value and softness inside the region of interest (ROI) were calculated. The correlations between variables were investigated and receiver operating characteristic (ROC) curve analysis was performed to assess the diagnostic accuracy of the proposed method. Pathologic results were used as standard reference. Results: Both the mean grey value and the softness inside the ROI resulted statistically different at the t test for the two populations of lesions (i.e., benign versus malignant): p < 0.0001. The area under the curve (AUC) was 0.924 (0.834–0.973) and 0.917 (0.826–0.970) for the mean grey value and for the softness respectively. Conclusions: Quantitative elastosonography is a promising ultrasound technique in the detection of breast cancer but large prospective trials are necessary to determine whether quantitative analysis of images can help to overcome some pitfalls of the methodic.

Renal geology (quantitative renal stone analysis) by 'Fourier transform infrared spectroscopy'.

Science.gov (United States)

Singh, Iqbal

2008-01-01

To prospectively determine the precise stone composition (quantitative analysis) by using infrared spectroscopy in patients with urinary stone disease presenting to our clinic. To determine an ideal method for stone analysis suitable for use in a clinical setting. After routine and a detailed metabolic workup of all patients of urolithiasis, stone samples of 50 patients of urolithiasis satisfying the entry criteria were subjected to the Fourier transform infrared spectroscopic analysis after adequate sample homogenization at a single testing center. Calcium oxalate monohydrate and dihydrate stone mixture was most commonly encountered in 35 (71%) followed by calcium phosphate, carbonate apatite, magnesium ammonium hexahydrate and xanthine stones. Fourier transform infrared spectroscopy allows an accurate, reliable quantitative method of stone analysis. It also helps in maintaining a computerized large reference library. Knowledge of precise stone composition may allow the institution of appropriate prophylactic therapy despite the absence of any detectable metabolic abnormalities. This may prevent and or delay stone recurrence.

Quantitative proteomic analysis of ibuprofen-degrading Patulibacter sp. strain I11

DEFF Research Database (Denmark)

Almeida, Barbara; Kjeldal, Henrik; Lolas, Ihab Bishara Yousef

2013-01-01

was identified and quantified by gel based shotgun-proteomics. In total 251 unique proteins were quantitated using this approach. Biological process and pathway analysis indicated a number of proteins that were up-regulated in response to active degradation of ibuprofen, some of them are known to be involved...... in the degradation of aromatic compounds. Data analysis revealed that several of these proteins are likely involved in ibuprofen degradation by Patulibacter sp. strain I11.......Ibuprofen is the third most consumed pharmaceutical drug in the world. Several isolates have been shown to degrade ibuprofen, but very little is known about the biochemistry of this process. This study investigates the degradation of ibuprofen by Patulibacter sp. strain I11 by quantitative...

Analysis association of milk fat and protein percent in quantitative ...

African Journals Online (AJOL)

Analysis association of milk fat and protein percent in quantitative trait locus ... African Journal of Biotechnology ... Protein and fat percent as content of milk are high-priority criteria for financial aims and selection of programs in dairy cattle.

Contribution of the surface contamination of uranium-materials on the quantitative analysis results by electron probe microbeam analysis

International Nuclear Information System (INIS)

Bonino, O.; Fournier, C.; Fucili, C.; Dugne, O.; Merlet, C.

2000-01-01

The analytical testing of uranium materials is necessary for quality research and development in nuclear industry applications (enrichment, safety studies, fuel, etc). Electron Probe Microbeam Analysis Wavelength Dispersive Spectrometry (EPMA-WDS) is a dependable non-destructive analytical technology. The characteristic X-ray signal is measured to identify and quantify the sample components, and the analyzed volume is about one micron cube. The surface contamination of uranium materials modifies and contributes to the quantitative analysis results of EPMA-WDS. This contribution is not representative of the bulk. A thin oxidized layer appears in the first instants after preparation (burnishing, cleaning) as well as a carbon contamination layer, due to metallographic preparation and carbon cracking under the impact of the electron probe. Several analytical difficulties subsequently arise, including an overlapping line between the carbon Ka ray and the Uranium U NIVOVI ray. Sensitivity and accuracy of the quantification of light elements like carbon and oxygen are also reduced by the presence of uranium. The aim of this study was to improve the accuracy of quantitative analysis on uranium materials by EPMA-WDS by taking account of the contribution of surface contamination. The first part of this paper is devoted to the study of the contaminated surface of the uranium materials U, UFe 2 and U 6 Fe a few hours after preparation. These oxidation conditions are selected so as to reproduce the same contamination surfaces occurring in microprobe analytical conditions. Surface characterization techniques were SIMS and Auger spectroscopy. The contaminated surfaces are shown. They consist of successive layers: a carbon layer, an oxidized iron layer, followed by an iron depletion layer (only in UFe 2 and U 6 Fe), and a ternary oxide layer (U-Fe-O for UFe 2 et U 6 Fe and UO 2+x for uranium). The second part of the paper addresses the estimation of the errors in quantitative

A new quantitative analysis on nitriding kinetics in the oxidized Zry-4 at 900-1200 .deg. C

Energy Technology Data Exchange (ETDEWEB)

Park, Sanggi [ACT Co. Ltd., Daejeon (Korea, Republic of)

2016-10-15

Two major roles of nitrogen on the zirconium based cladding degradation were identified: mechanical degradation of the cladding, and the additional chemical heat release. It has long been known that accelerated oxidation can occur in air due to the nitrogen. In addition, significant uptake of nitrogen can also occur. The nitriding of pre-oxidized zirconium based alloys leads to micro porous and less coherent oxide scales. This paper aims to quantitatively investigate the nitriding mechanism and kinetics by proposing a new methodology that is coupled with the mass balance analysis and the optical microscope image processing analysis. A new quantitative analysis methodology is described in chapter 2 and the investigation of the nitriding kinetics is performed in chapter 3. The experimental details are previously reported in. Previously only qualitative analysis was performed in, and hence the quantitative analysis will be performed in this paper. In this paper, the nitriding kinetics and mechanism were quantitatively analyzed by the new proposed analysis methods: the mass balance analysis and the optical microscope image processing analysis. Using these combined methods, the mass gain curves and the optical microscopes are analyzed in very detail, and the mechanisms of nitriding accelerated, stabilized and saturated behaviors were well understood. This paper has two very distinctive achievements as follows: 1) Development of very effective quantitative analysis methods only using two main results of oxidation tests: No detailed analytical sample measurements (e.g. TEM, EPMA and so on.) were required. These methods can effectively reduce the cost and effort of the post-test investigation. 2) The first identification of the nitriding behaviors and its very accurate analysis in a quantitative way. Based on this quantitative analysis results on the nitriding kinetics, these new findings will contribute significantly the understanding the air oxidation behaviors and model

Patient-specific coronary blood supply territories for quantitative perfusion analysis

Science.gov (United States)

Zakkaroff, Constantine; Biglands, John D.; Greenwood, John P.; Plein, Sven; Boyle, Roger D.; Radjenovic, Aleksandra; Magee, Derek R.

2018-01-01

Abstract Myocardial perfusion imaging, coupled with quantitative perfusion analysis, provides an important diagnostic tool for the identification of ischaemic heart disease caused by coronary stenoses. The accurate mapping between coronary anatomy and under-perfused areas of the myocardium is important for diagnosis and treatment. However, in the absence of the actual coronary anatomy during the reporting of perfusion images, areas of ischaemia are allocated to a coronary territory based on a population-derived 17-segment (American Heart Association) AHA model of coronary blood supply. This work presents a solution for the fusion of 2D Magnetic Resonance (MR) myocardial perfusion images and 3D MR angiography data with the aim to improve the detection of ischaemic heart disease. The key contribution of this work is a novel method for the mediated spatiotemporal registration of perfusion and angiography data and a novel method for the calculation of patient-specific coronary supply territories. The registration method uses 4D cardiac MR cine series spanning the complete cardiac cycle in order to overcome the under-constrained nature of non-rigid slice-to-volume perfusion-to-angiography registration. This is achieved by separating out the deformable registration problem and solving it through phase-to-phase registration of the cine series. The use of patient-specific blood supply territories in quantitative perfusion analysis (instead of the population-based model of coronary blood supply) has the potential of increasing the accuracy of perfusion analysis. Quantitative perfusion analysis diagnostic accuracy evaluation with patient-specific territories against the AHA model demonstrates the value of the mediated spatiotemporal registration in the context of ischaemic heart disease diagnosis. PMID:29392098

Unfolding of core nucleosomes by PARP-1 revealed by spFRET microscopy

Directory of Open Access Journals (Sweden)

Daniel Sultanov

2017-01-01

Full Text Available DNA accessibility to various protein complexes is essential for various processes in the cell and is affected by nucleosome structure and dynamics. Protein factor PARP-1 (poly(ADP-ribose polymerase 1 increases the accessibility of DNA in chromatin to repair proteins and transcriptional machinery, but the mechanism and extent of this chromatin reorganization are unknown. Here we report on the effects of PARP-1 on single nucleosomes revealed by spFRET (single-particle Förster Resonance Energy Transfer microscopy. PARP-1 binding to a double-strand break in the vicinity of a nucleosome results in a significant increase of the distance between the adjacent gyres of nucleosomal DNA. This partial uncoiling of the entire nucleosomal DNA occurs without apparent loss of histones and is reversed after poly(ADP-ribosylation of PARP-1. Thus PARP-1-nucleosome interactions result in reversible, partial uncoiling of the entire nucleosomal DNA.

Quantitative analysis of γ–oryzanol content in cold pressed rice bran oil by TLC–image analysis method

Directory of Open Access Journals (Sweden)

Apirak Sakunpak

2014-02-01

Conclusions: The TLC-densitometric and TLC-image analysis methods provided a similar reproducibility, accuracy and selectivity for the quantitative determination of γ-oryzanol in cold pressed rice bran oil. A statistical comparison of the quantitative determinations of γ-oryzanol in samples did not show any statistically significant difference between TLC-densitometric and TLC-image analysis methods. As both methods were found to be equal, they therefore can be used for the determination of γ-oryzanol in cold pressed rice bran oil.

Optimal climate policy is a utopia. From quantitative to qualitative cost-benefit analysis

International Nuclear Information System (INIS)

Van den Bergh, Jeroen C.J.M.

2004-01-01

The dominance of quantitative cost-benefit analysis (CBA) and optimality concepts in the economic analysis of climate policy is criticised. Among others, it is argued to be based in a misplaced interpretation of policy for a complex climate-economy system as being analogous to individual inter-temporal welfare optimisation. The transfer of quantitative CBA and optimality concepts reflects an overly ambitious approach that does more harm than good. An alternative approach is to focus the attention on extreme events, structural change and complexity. It is argued that a qualitative rather than a quantitative CBA that takes account of these aspects can support the adoption of a minimax regret approach or precautionary principle in climate policy. This means: implement stringent GHG reduction policies as soon as possible

Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)

DEFF Research Database (Denmark)

Skeldal, Sune; Kjaergaard, Maj M; Alwasel, Saleh

2015-01-01

the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay...

Method of quantitative analysis of superconducting metal-conducting composite materials

International Nuclear Information System (INIS)

Bogomolov, V.N.; Zhuravlev, V.V.; Petranovskij, V.P.; Pimenov, V.A.

1990-01-01

Technique for quantitative analysis of superconducting metal-containing composite materials, SnO 2 -InSn, WO 3 -InW, Zn)-InZn in particular, has been developed. The method of determining metal content in a composite is based on the dependence of superconducting transition temperature on alloy composition. Sensitivity of temperature determination - 0.02K, error of analysis for InSn system - 0.5%

Meta-analysis for quantitative microbiological risk assessments and benchmarking data

NARCIS (Netherlands)

Besten, den H.M.W.; Zwietering, M.H.

2012-01-01

Meta-analysis studies are increasingly being conducted in the food microbiology area to quantitatively integrate the findings of many individual studies on specific questions or kinetic parameters of interest. Meta-analyses provide global estimates of parameters and quantify their variabilities, and

Visual and Quantitative Analysis Methods of Respiratory Patterns for Respiratory Gated PET/CT.

Science.gov (United States)

Son, Hye Joo; Jeong, Young Jin; Yoon, Hyun Jin; Park, Jong-Hwan; Kang, Do-Young

2016-01-01

We integrated visual and quantitative methods for analyzing the stability of respiration using four methods: phase space diagrams, Fourier spectra, Poincaré maps, and Lyapunov exponents. Respiratory patterns of 139 patients were grouped based on the combination of the regularity of amplitude, period, and baseline positions. Visual grading was done by inspecting the shape of diagram and classified into two states: regular and irregular. Quantitation was done by measuring standard deviation of x and v coordinates of Poincaré map (SD x , SD v ) or the height of the fundamental peak ( A 1 ) in Fourier spectrum or calculating the difference between maximal upward and downward drift. Each group showed characteristic pattern on visual analysis. There was difference of quantitative parameters (SD x , SD v , A 1 , and MUD-MDD) among four groups (one way ANOVA, p = 0.0001 for MUD-MDD, SD x , and SD v , p = 0.0002 for A 1 ). In ROC analysis, the cutoff values were 0.11 for SD x (AUC: 0.982, p quantitative indices of respiratory stability and determining quantitative cutoff value for differentiating regular and irregular respiration.

GProX, a user-friendly platform for bioinformatics analysis and visualization of quantitative proteomics data.

Science.gov (United States)

Rigbolt, Kristoffer T G; Vanselow, Jens T; Blagoev, Blagoy

2011-08-01

Recent technological advances have made it possible to identify and quantify thousands of proteins in a single proteomics experiment. As a result of these developments, the analysis of data has become the bottleneck of proteomics experiment. To provide the proteomics community with a user-friendly platform for comprehensive analysis, inspection and visualization of quantitative proteomics data we developed the Graphical Proteomics Data Explorer (GProX)(1). The program requires no special bioinformatics training, as all functions of GProX are accessible within its graphical user-friendly interface which will be intuitive to most users. Basic features facilitate the uncomplicated management and organization of large data sets and complex experimental setups as well as the inspection and graphical plotting of quantitative data. These are complemented by readily available high-level analysis options such as database querying, clustering based on abundance ratios, feature enrichment tests for e.g. GO terms and pathway analysis tools. A number of plotting options for visualization of quantitative proteomics data is available and most analysis functions in GProX create customizable high quality graphical displays in both vector and bitmap formats. The generic import requirements allow data originating from essentially all mass spectrometry platforms, quantitation strategies and software to be analyzed in the program. GProX represents a powerful approach to proteomics data analysis providing proteomics experimenters with a toolbox for bioinformatics analysis of quantitative proteomics data. The program is released as open-source and can be freely downloaded from the project webpage at http://gprox.sourceforge.net.

Analysis of archaeological ceramics by total-reflection X-ray fluorescence: Quantitative approaches

International Nuclear Information System (INIS)

Fernandez-Ruiz, R.; Garcia-Heras, M.

2008-01-01

This paper reports the quantitative methodologies developed for the compositional characterization of archaeological ceramics by total-reflection X-ray fluorescence at two levels. A first quantitative level which comprises an acid leaching procedure, and a second selective level, which seeks to increase the number of detectable elements by eliminating the iron present in the acid leaching procedure. Total-reflection X-ray fluorescence spectrometry has been compared, at a quantitative level, with Instrumental Neutron Activation Analysis in order to test its applicability to the study of this kind of materials. The combination of a solid chemical homogenization procedure previously reported with the quantitative methodologies here presented allows the total-reflection X-ray fluorescence to analyze 29 elements with acceptable analytical recoveries and accuracies

Analysis of archaeological ceramics by total-reflection X-ray fluorescence: Quantitative approaches

Energy Technology Data Exchange (ETDEWEB)

Fernandez-Ruiz, R. [Servicio Interdepartamental de Investigacion, Facultad de Ciencias, Universidad Autonoma de Madrid, Modulo C-9, Laboratorio de TXRF, Crta. Colmenar, Km 15, Cantoblanco, E-28049, Madrid (Spain)], E-mail: ramon.fernandez@uam.es; Garcia-Heras, M. [Grupo de Arqueometria de Vidrios y Materiales Ceramicos, Instituto de Historia, Centro de Ciencias Humanas y Sociales, CSIC, C/ Albasanz, 26-28, 28037 Madrid (Spain)

2008-09-15

This paper reports the quantitative methodologies developed for the compositional characterization of archaeological ceramics by total-reflection X-ray fluorescence at two levels. A first quantitative level which comprises an acid leaching procedure, and a second selective level, which seeks to increase the number of detectable elements by eliminating the iron present in the acid leaching procedure. Total-reflection X-ray fluorescence spectrometry has been compared, at a quantitative level, with Instrumental Neutron Activation Analysis in order to test its applicability to the study of this kind of materials. The combination of a solid chemical homogenization procedure previously reported with the quantitative methodologies here presented allows the total-reflection X-ray fluorescence to analyze 29 elements with acceptable analytical recoveries and accuracies.

Study on methods of quantitative analysis of the biological thin samples in EM X-ray microanalysis

International Nuclear Information System (INIS)

Zhang Detian; Zhang Xuemin; He Kun; Yang Yi; Zhang Sa; Wang Baozhen

2000-01-01

Objective: To study the methods of quantitative analysis of the biological thin samples. Methods: Hall theory was used to study the qualitative analysis, background subtraction, peel off overlap peaks; external radiation and aberrance of spectra. Results: The results of reliable qualitative analysis and precise quantitative analysis were achieved. Conclusion: The methods for analysis of the biological thin samples in EM X-ray microanalysis can be used in biomedical research

Quantitative x-ray fractographic analysis of fatigue fractures

International Nuclear Information System (INIS)

Saprykin, Yu.V.

1983-01-01

The study deals with quantitative X-ray fractographic investigation of fatigue fractures of samples with sharp notches tested at various stresses and temperatures with the purpose of establishing a connection between material crack resistance parameters and local plastic instability zones restraining and controlling the crack growth. At fatigue fractures of notched Kh18N9T steel samples tested at +20 and -196 deg C a zone of sharp ring notch effect being analogous to the zone in which crack growth rate is controlled by the microshifting mechanisms is singled out. The size of the notched effect zone in the investigate steel is unambiguosly bound to to the stress amplitude. This provides the possibility to determine the stress value by the results of quantitative fractographic analysis of notched sample fractures. A possibility of determining one of the threshold values of cyclic material fracture toughness by the results of fatigue testing and fractography of notched sample fractures is shown. Correlation between the size of the hsub(s) crack effect zone in the notched sample, delta material yield limit and characteristic of cyclic Ksub(s) fracture toughness has been found. Such correlation widens the possibilities of quantitative diagnostics of fractures by the methods of X-ray fractography

Wavelength Selection Method Based on Differential Evolution for Precise Quantitative Analysis Using Terahertz Time-Domain Spectroscopy.

Science.gov (United States)

Li, Zhi; Chen, Weidong; Lian, Feiyu; Ge, Hongyi; Guan, Aihong

2017-12-01

Quantitative analysis of component mixtures is an important application of terahertz time-domain spectroscopy (THz-TDS) and has attracted broad interest in recent research. Although the accuracy of quantitative analysis using THz-TDS is affected by a host of factors, wavelength selection from the sample's THz absorption spectrum is the most crucial component. The raw spectrum consists of signals from the sample and scattering and other random disturbances that can critically influence the quantitative accuracy. For precise quantitative analysis using THz-TDS, the signal from the sample needs to be retained while the scattering and other noise sources are eliminated. In this paper, a novel wavelength selection method based on differential evolution (DE) is investigated. By performing quantitative experiments on a series of binary amino acid mixtures using THz-TDS, we demonstrate the efficacy of the DE-based wavelength selection method, which yields an error rate below 5%.

Feedback-mediated cancer therapy: a FRET-based nanoreporter approach

Science.gov (United States)

Sarkar, Suproteem K.; Khater, Yashika; Kulkarni, Ashish; Sengupta, Shiladitya

2014-08-01

A theranostic nanoparticle system was developed by integrating a chemotherapeutic agent with an "activatable" fluorescent tracer. The system signals tumor death by monitoring the activity of caspase-3, a product of apoptosis, and can therefore screen the treatment sensitivity of a particular tumor. The polymer nanoparticles (Poly [isobutylene-alt-maleic anhydride]) were formed through reprecipitation and contained paclitaxel, a chemotherapy drug, and fluorescein isothiocyanate, a fluorescent dye. The dye's fluorescence was quenched through Förster resonance energy transfer (FRET) by a quencher that was connected to the dye by a peptide chain. With sizes ranging from 200-250 nm, the nanoparticles were stable for two weeks. The nanoparticles were tested in vitro with responsive Lewis Lung Carcinoma (LLC) cells and taxane-resistant cells. Upon cell death by paclitaxel exposure, caspase-3 cleaved the peptide chain connecting the dye and the quencher, causing the system to fluoresce. When LLC cells were treated with the system, the nanoreporters fluoresced, but when resistant cells were tested, and when the drug was removed from the system, the nanoreporters did not fluoresce. Since the system screens if a drug can successfully kill a particular tumor, it offers a novel and promising approach to personalized medicine.

Quantitative texture analysis of electrodeposited line patterns

DEFF Research Database (Denmark)

Pantleon, Karen; Somers, Marcel A.J.

2005-01-01

Free-standing line patterns of Cu and Ni were manufactured by electrochemical deposition into lithographically prepared patterns. Electrodeposition was carried out on top of a highly oriented Au-layer physically vapor deposited on glass. Quantitative texture analysis carried out by means of x......-ray diffraction for both the substrate layer and the electrodeposits yielded experimental evidence for epitaxy between Cu and Au. An orientation relation between film and substrate was discussed with respect to various concepts of epitaxy. While the conventional mode of epitaxy fails for the Cu...

Joint analysis of binary and quantitative traits with data sharing and outcome-dependent sampling.

Science.gov (United States)

Zheng, Gang; Wu, Colin O; Kwak, Minjung; Jiang, Wenhua; Joo, Jungnam; Lima, Joao A C

2012-04-01

We study the analysis of a joint association between a genetic marker with both binary (case-control) and quantitative (continuous) traits, where the quantitative trait values are only available for the cases due to data sharing and outcome-dependent sampling. Data sharing becomes common in genetic association studies, and the outcome-dependent sampling is the consequence of data sharing, under which a phenotype of interest is not measured for some subgroup. The trend test (or Pearson's test) and F-test are often, respectively, used to analyze the binary and quantitative traits. Because of the outcome-dependent sampling, the usual F-test can be applied using the subgroup with the observed quantitative traits. We propose a modified F-test by also incorporating the genotype frequencies of the subgroup whose traits are not observed. Further, a combination of this modified F-test and Pearson's test is proposed by Fisher's combination of their P-values as a joint analysis. Because of the correlation of the two analyses, we propose to use a Gamma (scaled chi-squared) distribution to fit the asymptotic null distribution for the joint analysis. The proposed modified F-test and the joint analysis can also be applied to test single trait association (either binary or quantitative trait). Through simulations, we identify the situations under which the proposed tests are more powerful than the existing ones. Application to a real dataset of rheumatoid arthritis is presented. © 2012 Wiley Periodicals, Inc.

Investment appraisal using quantitative risk analysis.

Science.gov (United States)

Johansson, Henrik

2002-07-01

Investment appraisal concerned with investments in fire safety systems is discussed. Particular attention is directed at evaluating, in terms of the Bayesian decision theory, the risk reduction that investment in a fire safety system involves. It is shown how the monetary value of the change from a building design without any specific fire protection system to one including such a system can be estimated by use of quantitative risk analysis, the results of which are expressed in terms of a Risk-adjusted net present value. This represents the intrinsic monetary value of investing in the fire safety system. The method suggested is exemplified by a case study performed in an Avesta Sheffield factory.

Nanophotonic Control of the Förster Resonance Energy Transfer Efficiency

DEFF Research Database (Denmark)

Blum, Christian; Zijlstra, Niels; Lagendijk, Ad

2012-01-01

We have studied the influence of the local density of optical states (LDOS) on the rate and efficiency of Forster resonance energy transfer (FRET) from a donor to an acceptor. The donors and acceptors are dye molecules that are separated by a short strand of double-stranded DNA. The LDOS...... is controlled by carefully positioning the FRET pairs near a mirror. We find that the energy transfer efficiency changes with LDOS, and that, in agreement with theory, the energy transfer rate is independent of the LDOS, which allows one to quantitatively control FRET systems in a new way. Our results imply...

Subunit rotation in a single FoF1-ATP synthase in a living bacterium monitored by FRET

Science.gov (United States)

Seyfert, K.; Oosaka, T.; Yaginuma, H.; Ernst, S.; Noji, H.; Iino, R.; Börsch, M.

2011-03-01

FoF1-ATP synthase is the ubiquitous membrane-bound enzyme in mitochondria, chloroplasts and bacteria which provides the 'chemical energy currency' adenosine triphosphate (ATP) for cellular processes. In Escherichia coli ATP synthesis is driven by a proton motive force (PMF) comprising a proton concentration difference ΔpH plus an electric potential ΔΨ across the lipid membrane. Single-molecule in vitro experiments have confirmed that proton-driven subunit rotation within FoF1-ATP synthase is associated with ATP synthesis. Based on intramolecular distance measurements by single-molecule fluorescence resonance energy transfer (FRET) the kinetics of subunit rotation and the step sizes of the different rotor parts have been unraveled. However, these experiments were accomplished in the presence of a PMF consisting of a maximum ΔpH ~ 4 and an unknown ΔΨ. In contrast, in living bacteria the maximum ΔpH across the plasma membrane is likely 0.75, and ΔΨ has been measured between -80 and -140 mV. Thus the problem of in vivo catalytic turnover rates, or the in vivo rotational speed in single FoF1-ATP synthases, respectively, has to be solved. In addition, the absolute number of functional enzymes in a single bacterium required to maintain the high ATP levels has to be determined. We report our progress of measuring subunit rotation in single FoF1-ATP synthases in vitro and in vivo, which was enabled by a new labeling approach for single-molecule FRET measurements.

GProX, a User-Friendly Platform for Bioinformatics Analysis and Visualization of Quantitative Proteomics Data

DEFF Research Database (Denmark)

Rigbolt, Kristoffer T G; Vanselow, Jens T; Blagoev, Blagoy

2011-01-01

-friendly platform for comprehensive analysis, inspection and visualization of quantitative proteomics data we developed the Graphical Proteomics Data Explorer (GProX)(1). The program requires no special bioinformatics training, as all functions of GProX are accessible within its graphical user-friendly interface...... such as database querying, clustering based on abundance ratios, feature enrichment tests for e.g. GO terms and pathway analysis tools. A number of plotting options for visualization of quantitative proteomics data is available and most analysis functions in GProX create customizable high quality graphical...... displays in both vector and bitmap formats. The generic import requirements allow data originating from essentially all mass spectrometry platforms, quantitation strategies and software to be analyzed in the program. GProX represents a powerful approach to proteomics data analysis providing proteomics...

Survival Prediction in Pancreatic Ductal Adenocarcinoma by Quantitative Computed Tomography Image Analysis.

Science.gov (United States)

Attiyeh, Marc A; Chakraborty, Jayasree; Doussot, Alexandre; Langdon-Embry, Liana; Mainarich, Shiana; Gönen, Mithat; Balachandran, Vinod P; D'Angelica, Michael I; DeMatteo, Ronald P; Jarnagin, William R; Kingham, T Peter; Allen, Peter J; Simpson, Amber L; Do, Richard K

2018-04-01

Pancreatic cancer is a highly lethal cancer with no established a priori markers of survival. Existing nomograms rely mainly on post-resection data and are of limited utility in directing surgical management. This study investigated the use of quantitative computed tomography (CT) features to preoperatively assess survival for pancreatic ductal adenocarcinoma (PDAC) patients. A prospectively maintained database identified consecutive chemotherapy-naive patients with CT angiography and resected PDAC between 2009 and 2012. Variation in CT enhancement patterns was extracted from the tumor region using texture analysis, a quantitative image analysis tool previously described in the literature. Two continuous survival models were constructed, with 70% of the data (training set) using Cox regression, first based only on preoperative serum cancer antigen (CA) 19-9 levels and image features (model A), and then on CA19-9, image features, and the Brennan score (composite pathology score; model B). The remaining 30% of the data (test set) were reserved for independent validation. A total of 161 patients were included in the analysis. Training and test sets contained 113 and 48 patients, respectively. Quantitative image features combined with CA19-9 achieved a c-index of 0.69 [integrated Brier score (IBS) 0.224] on the test data, while combining CA19-9, imaging, and the Brennan score achieved a c-index of 0.74 (IBS 0.200) on the test data. We present two continuous survival prediction models for resected PDAC patients. Quantitative analysis of CT texture features is associated with overall survival. Further work includes applying the model to an external dataset to increase the sample size for training and to determine its applicability.

Limitations for qualitative and quantitative neutron activation analysis using reactor neutrons

International Nuclear Information System (INIS)

El-Abbady, W.H.; El-Tanahy, Z.H.; El-Hagg, A.A.; Hassan, A.M.

1999-01-01

In this work, the most important limitations for qualitative and quantitative analysis using reactor neutrons for activation are reviewed. Each limitation is discussed using different examples of activated samples. Photopeak estimation, nuclear reactions interference and neutron flux measurements are taken into consideration. Solutions for high accuracy evaluation in neutron activation analysis applications are given. (author)

From Dark to Light to Fluorescence Resonance Energy Transfer (FRET): Polarity-Sensitive Aggregation-Induced Emission (AIE)-Active Tetraphenylethene-Fused BODIPY Dyes with a Very Large Pseudo-Stokes Shift.

Science.gov (United States)

Şen, Esra; Meral, Kadem; Atılgan, Serdar

2016-01-11

The work presented herein is devoted to the fabrication of large Stokes shift dyes in both organic and aqueous media by combining dark resonance energy transfer (DRET) and fluorescence resonance energy transfer (FRET) in one donor-acceptor system. In this respect, a series of donor-acceptor architectures of 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) dyes substituted by one, two, or three tetraphenylethene (TPE) luminogens were designed and synthesised. The photophysical properties of these three chromophore systems were studied to provide insight into the nature of donor-acceptor interactions in both THF and aqueous media. Because the generation of emissive TPE donor(s) is strongly polarity dependent, due to its aggregation-induced emission (AIE) feature, one might expect the formation of appreciable fluorescence emission intensity with a very large pseudo-Stokes shift in aqueous media when considering FRET process. Interestingly, similar results were also recorded in THF for the chromophore systems, although the TPE fragment(s) of the dyes are non-emissive. The explanation for this photophysical behaviour lies in the DRET. This is the first report on combining two energy-transfer processes, namely, FRET and DRET, in one polarity-sensitive donor-acceptor pair system. The accuracy of the dark-emissive donor property of the TPE luminogen is also presented for the first time as a new feature for AIE phenomena. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Program for the quantitative and qualitative analysis of

International Nuclear Information System (INIS)

Tepelea, V.; Purice, E.; Dan, R.; Calcev, G.; Domnisan, M.; Galis, V.; Teodosiu, G.; Debert, C.; Mocanu, N.; Nastase, M.

1985-01-01

A computer code for processing of data from neutron activation analysis is described. The code is capable of qualitative and quantitative analysis of regular spectra from neutron irradiated samples, measured by a Ge(li) detector. Multichannel analysers with 1024 channels, such as TN 1705 or a Romanian made MCA 79, and an ITC interface can be used. The code is implemented on FELIX M118 and FELIX M216 microcomputers. Spectrum processing is performed off line, after storing the data on a floppy disk. The background is assumed to be a polynomial of first, second or third degree. Qualitative analysis is performed by recursive least square, Gaussian curve fitting. The elements are identified using a polynomial relation between energy and channel, obtained by calibration with a standard sample

Multivariate analysis of quantitative traits can effectively classify rapeseed germplasm

Directory of Open Access Journals (Sweden)

Jankulovska Mirjana

2014-01-01

Full Text Available In this study, the use of different multivariate approaches to classify rapeseed genotypes based on quantitative traits has been presented. Tree regression analysis, PCA analysis and two-way cluster analysis were applied in order todescribe and understand the extent of genetic variability in spring rapeseed genotype by trait data. The traits which highly influenced seed and oil yield in rapeseed were successfully identified by the tree regression analysis. Principal predictor for both response variables was number of pods per plant (NP. NP and 1000 seed weight could help in the selection of high yielding genotypes. High values for both traits and oil content could lead to high oil yielding genotypes. These traits may serve as indirect selection criteria and can lead to improvement of seed and oil yield in rapeseed. Quantitative traits that explained most of the variability in the studied germplasm were classified using principal component analysis. In this data set, five PCs were identified, out of which the first three PCs explained 63% of the total variance. It helped in facilitating the choice of variables based on which the genotypes’ clustering could be performed. The two-way cluster analysissimultaneously clustered genotypes and quantitative traits. The final number of clusters was determined using bootstrapping technique. This approach provided clear overview on the variability of the analyzed genotypes. The genotypes that have similar performance regarding the traits included in this study can be easily detected on the heatmap. Genotypes grouped in the clusters 1 and 8 had high values for seed and oil yield, and relatively short vegetative growth duration period and those in cluster 9, combined moderate to low values for vegetative growth duration and moderate to high seed and oil yield. These genotypes should be further exploited and implemented in the rapeseed breeding program. The combined application of these multivariate methods