WorldWideScience

Sample records for greater fret values

  1. Greater effort increases perceived value in an invertebrate.

    Science.gov (United States)

    Czaczkes, Tomer J; Brandstetter, Birgit; di Stefano, Isabella; Heinze, Jürgen

    2018-05-01

    Expending effort is generally considered to be undesirable. However, both humans and vertebrates will work for a reward they could also get for free. Moreover, cues associated with high-effort rewards are preferred to low-effort associated cues. Many explanations for these counterintuitive findings have been suggested, including cognitive dissonance (self-justification) or a greater contrast in state (e.g., energy or frustration level) before and after an effort-linked reward. Here, we test whether effort expenditure also increases perceived value in ants, using both classical cue-association methods and pheromone deposition, which correlates with perceived value. In 2 separate experimental setups, we show that pheromone deposition is higher toward the reward that requires more effort: 47% more pheromone deposition was performed for rewards reached via a vertical runway (high effort) compared with ones reached via a horizontal runway (low effort), and deposition rates were 28% higher on rough (high effort) versus smooth (low effort) runways. Using traditional cue-association methods, 63% of ants trained on different surface roughness, and 70% of ants trained on different runway elevations, preferred the high-effort related cues on a Y maze. Finally, pheromone deposition to feeders requiring memorization of one path bifurcation was up to 29% higher than to an identical feeder requiring no learning. Our results suggest that effort affects value perception in ants. This effect may stem from a cognitive process, which monitors the change in a generalized hedonic state before and after reward. (PsycINFO Database Record (c) 2018 APA, all rights reserved).

  2. More Value through Greater Differentiation: Gender Differences in Value Beliefs about Math

    Science.gov (United States)

    Gaspard, Hanna; Dicke, Anna-Lena; Flunger, Barbara; Schreier, Brigitte; Häfner, Isabelle; Trautwein, Ulrich; Nagengast, Benjamin

    2015-01-01

    Expectancy-value theory (Eccles et al., 1983) is a prominent approach to explaining gender differences in math-related academic choices, with value beliefs acting as an important explanatory factor. Expectancy-value theory defines 4 value components: intrinsic value, attainment value, utility value, and cost. The present study followed up on…

  3. 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.

  4. Call to action: Better care, better health, and greater value in college health.

    Science.gov (United States)

    Ciotoli, Carlo; Smith, Allison J; Keeling, Richard P

    2018-03-05

    It is time for action by leaders across higher education to strengthen quality improvement (QI) in college health, in pursuit of better care, better health, and increased value - goals closely linked to students' learning and success. The size and importance of the college student population; the connections between wellbeing, and therefore QI, and student success; the need for improved standards and greater accountability; and the positive contributions of QI to employee satisfaction and professionalism all warrant a widespread commitment to building greater capacity and capability for QI in college health. This report aims to inspire, motivate, and challenge college health professionals and their colleagues, campus leaders, and national entities to take both immediate and sustainable steps to bring QI to the forefront of college health practice - and, by doing so, to elevate care, health, and value of college health as a key pathway to advancing student success.

  5. The Effects of on Blood Glucose Values are Greater than those of Dietary Changes Alone

    Directory of Open Access Journals (Sweden)

    Ashley N. Hoehn

    2012-01-01

    Full Text Available Eighteen type II diabetics (9 women and 9 men participated in a 12-week trial that consisted of 2 parts, a 3-week control phase followed by a 9-week experimental phase where half of the subjects received 1000 mg of Cinnamomum cassia while the other half received 1000 mg of a placebo pill. All of the subjects that were in the cinnamon group had a statistically significant decrease in their blood sugar levels with a P -value of 3.915 × 10 -10 . The subjects in the cinnamon group had an average overall decrease in their blood sugar levels of about 30 mg/dL, which is comparable to oral medications available for diabetes. All subjects were educated on appropriate diabetic diets and maintained that diet for the entire 12 week study. Greater decreases in blood glucose values were observed in patients using the cinnamon compared to those using the dietary changes alone.

  6. Work stress, Chinese work values, and work well-being in the Greater China.

    Science.gov (United States)

    Lu, Luo; Kao, Shu-Fang; Siu, Oi-Ling; Lu, Chang-Qin

    2011-01-01

    Work values influence our attitudes and behavior at work, but they have rarely been explored in the context of work stress. The aim of this research was thus to test direct and moderating effects of Chinese work values (CWV) on relationships between work stressors and work well-being among employees in the Greater China region. A self-administered survey was conducted to collect data from three major cities in the region, namely Beijing, Hong Kong, and Taipei (N = 380). Work stressors were negatively related to work well-being, whereas CWV were positively related to work well-being. In addition, CWV also demonstrated moderating effects in some of the stressor-job satisfaction relationships.

  7. 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...

  8. Greater decision-making competence is associated with greater expected-value sensitivity, but not overall risk taking: An examination of concurrent validity

    Directory of Open Access Journals (Sweden)

    Andrew M Parker

    2015-05-01

    Full Text Available Decision-making competence reflects individual differences in the susceptibility to decision-making errors, measured using tasks common from behavioral decision research (e.g., framing effects, under/overconfidence, following decision rules. Prior research demonstrates that those with higher decision-making competence report lower incidence of health-risking and antisocial behaviors, but there has been less focus on intermediate mechanisms that may impact real-world decisions, and, in particular, those implicated by normative models. Here we test the associations between measures of youth decision-making competence (Y-DMC and one such mechanism, the degree to which individuals make choices consistent with maximizing expected value (EV. Using a task involving hypothetical gambles, we find that greater EV sensitivity is associated with greater Y-DMC. Higher Y-DMC scores are associated with (a choosing risky options when expected value favors those options and (b avoiding risky options when expected value favors a certain option. This relationship is stronger for gambles that involved potential losses. The results suggest that Y-DMC captures decision processes consistent with standard normative evaluations of risky decisions.

  9. Characteristic of fretting damage in metal material

    Energy Technology Data Exchange (ETDEWEB)

    Li, D.; Zhi, F.

    1988-10-01

    The fretting fatigue experiment of LC4 high strength aluminum alloy is described. An SEM examination of the fractology and morphology of fretting damage is carried out as well as an EDAX analysis of the chemical composition of fretting particles. The results show that many loose oxide particles were produced and accumulated in the fretting damage region. 10 references.

  10. Greater decision-making competence is associated with greater expected-value sensitivity, but not overall risk taking: an examination of concurrent validity.

    Science.gov (United States)

    Parker, Andrew M; Weller, Joshua A

    2015-01-01

    Decision-making competence reflects individual differences in the susceptibility to committing decision-making errors, measured using tasks common from behavioral decision research (e.g., framing effects, under/overconfidence, following decision rules). Prior research demonstrates that those with higher decision-making competence report lower incidence of health-risking and antisocial behaviors, but there has been less focus on intermediate processes that may impact real-world decisions, and, in particular, those implicated by normative models. Here we test the associations between measures of youth decision-making competence (Y-DMC) and one such process, the degree to which individuals make choices consistent with maximizing expected value (EV). Using a task involving hypothetical gambles, we find that greater EV sensitivity is associated with greater Y-DMC. Higher Y-DMC scores are associated with (a) choosing risky options when EV favors those options and (b) avoiding risky options when EV favors a certain option. This relationship is stronger for gambles that involved potential losses. The results suggest that Y-DMC captures decision processes consistent with standard normative evaluations of risky decisions.

  11. Innovative Payment Solutions in Agricultural Value Chain as a Means for Greater Financial Inclusion

    OpenAIRE

    Pandey, Tushar; Krishna, Nagahari; Vickers, Venetia; Menezes, Antonio; Raghavendra, M.

    2010-01-01

    The efforts for financial inclusion need to be designed with a vision beyond just the percentage of the country population with access to a bank account or a no frills account; to focus more on how this can enhance the capability and convenience for the un-banked and under-banked, specifically the small and marginal farmers in this case, to enable greater transparency, accountability, efficiency and convenient access to necessary facilities. The growth of ICT industry and mobile telecom revol...

  12. Improving Indonesian cinnamon (c. burmannii (Nees & t. nees) Blume) value chains for Greater Farmers Incomes

    Science.gov (United States)

    Menggala, S. R.; Damme, P. V.

    2018-03-01

    Genus Cinnamomum (Lauraceae) regroups some species whose stem bark are harvested, conditioned and traded as cinnamon in an international market. Over the centuries, the species have been domesticated so that now at least six different ones are grown in Southeast Asia countries. One of the species is Cinnamomum burmannii, also known as Korintje Cinnamon, which generates income for most smallholder farmers in Kerinci district, Jambi, Indonesia. Most cinnamon consumed in the world originates from this Korintje Cinnamon products. It is recognized for its unparalleled quality that comes with its sharp and sweet flavor, with a slightly bitter edge. However, international market requirements for product certification and quality standards make it difficult for a farmer to comply. Our research will address issues related to (improvement of) productivity, sustainability and value chains faced by cinnamon producers in Kerinci, to strengthen their product’s value chains. Smallholder farmers are very vulnerable to climate change impacts, and thus empowering the value chains of agricultural products will increase farmers resilience to climate change. The research will analyze the development of agricultural value chains, certification & standards on trade mechanism to help farmers earn a better income and future prospects.

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. A greater decline in female facial attractiveness during middle age reflects women’s loss of reproductive value

    Directory of Open Access Journals (Sweden)

    Dario eMaestripieri

    2014-02-01

    Full Text Available Facial attractiveness represents an important component of an individual’s overall attractiveness as a potential mating partner. Perceptions of facial attractiveness are expected to vary with age-related changes in health, reproductive value, and power. In this study, we investigated perceptions of facial attractiveness, power, and personality in two groups of women of pre- and post-menopausal ages (35-50 years and 51-65 years, respectively and two corresponding groups of men. We tested three hypotheses: 1 that perceived facial attractiveness would be lower for older than for younger men and women; 2 that the age-related reduction in facial attractiveness would be greater for women than for men; and 3 that for men, there would be a larger increase in perceived power at older ages. Eighty facial stimuli were rated by 60 (30 male, 30 female middle-aged women and men using online surveys. Our three main hypotheses were supported by the data. Consistent with sex differences in mating strategies, the greater age-related decline in female facial attractiveness was driven by male respondents, while the greater age-related increase in male perceived power was driven by female respondents. In addition, we found evidence that some personality ratings were correlated with perceived attractiveness and power ratings. The results of this study are consistent with evolutionary theory and with previous research showing that faces can provide important information about characteristics that men and women value in a potential mating partner such as their health, reproductive value, and power or possession of resources.

  18. 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.

  19. 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.

  20. 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.

  1. 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.

  2. 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...

  3. 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.

  4. 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

  5. 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...

  6. 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.

  7. 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...

  8. Evolving reimbursement and pricing policies for devices in Europe and the United States should encourage greater value.

    Science.gov (United States)

    Sorenson, Corinna; Drummond, Michael; Burns, Lawton R

    2013-04-01

    Rising health care costs are an international concern, particularly in the United States, where spending on health care outpaces that of other industrialized countries. Consequently, there is growing desire in the United States and Europe to take a more value-based approach to health care, particularly with respect to the adoption and use of new health technology. This article examines medical device reimbursement and pricing policies in the United States and Europe, with a particular focus on value. Compared to the United States, Europe more formally and consistently considers value to determine which technologies to cover and at what price, especially for complex, costly devices. Both the United States and Europe have introduced policies to provide temporary coverage and reimbursement for promising technologies while additional evidence of value is generated. But additional actions are needed in both the United States and Europe to ensure wise value-based reimbursement and pricing policies for all devices, including the generation of better pre- and postmarket evidence and the development of new methods to evaluate value and link evidence of value to reimbursement.

  9. The Economic Value of the Greater Montreal Blue Network (Quebec, Canada: A Contingent Choice Study Using Real Projects to Estimate Non-Market Aquatic Ecosystem Services Benefits.

    Directory of Open Access Journals (Sweden)

    Thomas G Poder

    Full Text Available This study used a contingent choice method to determine the economic value of improving various ecosystem services (ESs of the Blue Network of Greater Montreal (Quebec, Canada. Three real projects were used and the evaluation focused on six ESs that are related to freshwater aquatic ecosystems: biodiversity, water quality, carbon sequestration, recreational activities, landscape aesthetics and education services. We also estimated the value associated with the superficies of restored sites. We calculated the monetary value that a household would be willing to pay for each additional qualitative or quantitative unit of different ESs, and these marginal values range from $0.11 to $15.39 per household per unit. Thus, under certain assumptions, we determined the monetary values that all Quebec households would allocate to improve each ES in Greater Montreal by one unit. The most valued ES was water quality ($13.5 million, followed by education services ($10.7 million, recreational activities ($8.9 million, landscape aesthetics ($4.1 million, biodiversity ($1.2 million, and carbon sequestration ($0.1 million. Our results ascribe monetary values to improved (or degraded aquatic ecosystems in the Blue Network of Greater Montreal, but can also enhance economic analyses of various aquatic ecosystem restoration and management projects.

  10. The Economic Value of the Greater Montreal Blue Network (Quebec, Canada): A Contingent Choice Study Using Real Projects to Estimate Non-Market Aquatic Ecosystem Services Benefits.

    Science.gov (United States)

    Poder, Thomas G; Dupras, Jérôme; Fetue Ndefo, Franck; He, Jie

    2016-01-01

    This study used a contingent choice method to determine the economic value of improving various ecosystem services (ESs) of the Blue Network of Greater Montreal (Quebec, Canada). Three real projects were used and the evaluation focused on six ESs that are related to freshwater aquatic ecosystems: biodiversity, water quality, carbon sequestration, recreational activities, landscape aesthetics and education services. We also estimated the value associated with the superficies of restored sites. We calculated the monetary value that a household would be willing to pay for each additional qualitative or quantitative unit of different ESs, and these marginal values range from $0.11 to $15.39 per household per unit. Thus, under certain assumptions, we determined the monetary values that all Quebec households would allocate to improve each ES in Greater Montreal by one unit. The most valued ES was water quality ($13.5 million), followed by education services ($10.7 million), recreational activities ($8.9 million), landscape aesthetics ($4.1 million), biodiversity ($1.2 million), and carbon sequestration ($0.1 million). Our results ascribe monetary values to improved (or degraded) aquatic ecosystems in the Blue Network of Greater Montreal, but can also enhance economic analyses of various aquatic ecosystem restoration and management projects.

  11. 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

  12. 12 CFR 714.5 - What is required if you rely on an estimated residual value greater than 25% of the original cost...

    Science.gov (United States)

    2010-01-01

    ... 12 Banks and Banking 6 2010-01-01 2010-01-01 false What is required if you rely on an estimated residual value greater than 25% of the original cost of the leased property? 714.5 Section 714.5 Banks and Banking NATIONAL CREDIT UNION ADMINISTRATION REGULATIONS AFFECTING CREDIT UNIONS LEASING § 714.5 What is...

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. Simulation of FRET dyes allows quantitative comparison against experimental data

    Science.gov (United States)

    Reinartz, Ines; Sinner, Claude; Nettels, Daniel; Stucki-Buchli, Brigitte; Stockmar, Florian; Panek, Pawel T.; Jacob, Christoph R.; Nienhaus, Gerd Ulrich; Schuler, Benjamin; Schug, Alexander

    2018-03-01

    Fully understanding biomolecular function requires detailed insight into the systems' structural dynamics. Powerful experimental techniques such as single molecule Förster Resonance Energy Transfer (FRET) provide access to such dynamic information yet have to be carefully interpreted. Molecular simulations can complement these experiments but typically face limits in accessing slow time scales and large or unstructured systems. Here, we introduce a coarse-grained simulation technique that tackles these challenges. While requiring only few parameters, we maintain full protein flexibility and include all heavy atoms of proteins, linkers, and dyes. We are able to sufficiently reduce computational demands to simulate large or heterogeneous structural dynamics and ensembles on slow time scales found in, e.g., protein folding. The simulations allow for calculating FRET efficiencies which quantitatively agree with experimentally determined values. By providing atomically resolved trajectories, this work supports the planning and microscopic interpretation of experiments. Overall, these results highlight how simulations and experiments can complement each other leading to new insights into biomolecular dynamics and function.

  18. 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.

  19. 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

  20. 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.

  1. Parallel multispot smFRET analysis using an 8-pixel SPAD array

    Science.gov (United States)

    Ingargiola, A.; Colyer, R. A.; Kim, D.; Panzeri, F.; Lin, R.; Gulinatti, A.; Rech, I.; Ghioni, M.; Weiss, S.; Michalet, X.

    2012-02-01

    Single-molecule Förster resonance energy transfer (smFRET) is a powerful tool for extracting distance information between two fluorophores (a donor and acceptor dye) on a nanometer scale. This method is commonly used to monitor binding interactions or intra- and intermolecular conformations in biomolecules freely diffusing through a focal volume or immobilized on a surface. The diffusing geometry has the advantage to not interfere with the molecules and to give access to fast time scales. However, separating photon bursts from individual molecules requires low sample concentrations. This results in long acquisition time (several minutes to an hour) to obtain sufficient statistics. It also prevents studying dynamic phenomena happening on time scales larger than the burst duration and smaller than the acquisition time. Parallelization of acquisition overcomes this limit by increasing the acquisition rate using the same low concentrations required for individual molecule burst identification. In this work we present a new two-color smFRET approach using multispot excitation and detection. The donor excitation pattern is composed of 4 spots arranged in a linear pattern. The fluorescent emission of donor and acceptor dyes is then collected and refocused on two separate areas of a custom 8-pixel SPAD array. We report smFRET measurements performed on various DNA samples synthesized with various distances between the donor and acceptor fluorophores. We demonstrate that our approach provides identical FRET efficiency values to a conventional single-spot acquisition approach, but with a reduced acquisition time. Our work thus opens the way to high-throughput smFRET analysis on freely diffusing molecules.

  2. 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

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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.)

  8. 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.

  9. 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

  10. 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

  11. 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

  12. 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.

  13. 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 ...

  14. 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.

  15. 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)

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. A new trend to determine biochemical parameters by quantitative FRET assays.

    Science.gov (United States)

    Liao, Jia-yu; Song, Yang; Liu, Yan

    2015-12-01

    Förster resonance energy transfer (FRET) has been widely used in biological and biomedical research because it can determine molecule or particle interactions within a range of 1-10 nm. The sensitivity and efficiency of FRET strongly depend on the distance between the FRET donor and acceptor. Historically, FRET assays have been used to quantitatively deduce molecular distances. However, another major potential application of the FRET assay has not been fully exploited, that is, the use of FRET signals to quantitatively describe molecular interactive events. In this review, we discuss the use of quantitative FRET assays for the determination of biochemical parameters, such as the protein interaction dissociation constant (K(d)), enzymatic velocity (k(cat)) and K(m). We also describe fluorescent microscopy-based quantitative FRET assays for protein interaction affinity determination in cells as well as fluorimeter-based quantitative FRET assays for protein interaction and enzymatic parameter determination in solution.

  1. In vivo dynamics of enterovirus protease revealed by fluorescence resonance emission transfer (FRET) based on a novel FRET pair

    International Nuclear Information System (INIS)

    Hsu, Y.-Y.; Liu, Y.-N.; Wang Wenyen; Kao, Fu-Jen; Kung, S.-H.

    2007-01-01

    An in vivo protease assay suitable for analysis by fluorescence resonance energy transfer (FRET) was developed on the basis of a novel FRET pair. The specifically designed fusion substrate consists of green fluorescent protein 2 (GFP 2 )-peptide-red fluorescent protein 2 (DsRed2), with a cleavage motif for the enterovirus 2A protease (2A pro ) embedded within the peptide region. FRET can be readily visualized in real-time from cells expressing the fusion substrate until a proteolytic cleavage by 2A pro from the input virus. The level of FRET decay is a function of the amount and infection duration of the inoculated virus as measured by a fluorometer assay. The FRET biosensor also responded well to other related enteroviruses but not to a phylogenetically distant virus. Western blot analysis confirmed the physical cleavage of the fusion substrate upon the infections. The study provides proof of principle for applying the FRET technology to diagnostics, screening procedures, and cell biological research

  2. 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.

  3. 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

  4. 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.

  5. 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

  6. 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...

  7. 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.

  8. 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

  9. Effect of recycling activities on the heating value of solid waste: case study of the Greater Vancouver Regional District (Metro Vancouver).

    Science.gov (United States)

    Abedini, Ali R; Atwater, James W; Fu, George Yuzhu

    2012-08-01

    Two main goals of the integrated solid waste management system (ISWMS) of Metro Vancouver (MV) include further recycling of waste and energy recovery via incineration of waste. These two very common goals, however, are not always compatible enough to fit in an ISWMS depending on waste characteristics and details of recycling programs. This study showed that recent recycling activities in MV have negatively affected the net heating value (NHV) of municipal solid waste (MSW) in this regional district. Results show that meeting MV's goal for additional recycling of MSW by 2015 will further reduce the NHV of waste, if additional recycling activities are solely focused on more extensive recycling of packaging materials (e.g. paper and plastic). It is concluded that 50% additional recycling of paper and plastic in MV will increase the overall recycling rate to 70% (as targeted by the MV for 2015) and result in more than 8% reduction in NHV of MSW. This reduction translates to up to 2.3 million Canadian dollar (CAD$) less revenue at a potential waste-to-energy (WTE) plant with 500 000 tonnes year(-1) capacity. Properly designed recycling programmes, however, can make this functional element of ISWMS compatible with green goals of energy recovery from waste. Herein an explanation of how communities can increase their recycling activities without affecting the feasibility of potential WTE projects is presented.

  10. 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)

  11. 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.

  12. 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.

  13. Characterizing single-molecule FRET dynamics with probability distribution analysis.

    Science.gov (United States)

    Santoso, Yusdi; Torella, Joseph P; Kapanidis, Achillefs N

    2010-07-12

    Probability distribution analysis (PDA) is a recently developed statistical tool for predicting the shapes of single-molecule fluorescence resonance energy transfer (smFRET) histograms, which allows the identification of single or multiple static molecular species within a single histogram. We used a generalized PDA method to predict the shapes of FRET histograms for molecules interconverting dynamically between multiple states. This method is tested on a series of model systems, including both static DNA fragments and dynamic DNA hairpins. By fitting the shape of this expected distribution to experimental data, the timescale of hairpin conformational fluctuations can be recovered, in good agreement with earlier published results obtained using different techniques. This method is also applied to studying the conformational fluctuations in the unliganded Klenow fragment (KF) of Escherichia coli DNA polymerase I, which allows both confirmation of the consistency of a simple, two-state kinetic model with the observed smFRET distribution of unliganded KF and extraction of a millisecond fluctuation timescale, in good agreement with rates reported elsewhere. We expect this method to be useful in extracting rates from processes exhibiting dynamic FRET, and in hypothesis-testing models of conformational dynamics against experimental data.

  14. 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

  15. Uncovering Aberrant Mutant PKA Function with Flow Cytometric FRET

    Directory of Open Access Journals (Sweden)

    Shin-Rong Lee

    2016-03-01

    Full Text Available Biology has been revolutionized by tools that allow the detection and characterization of protein-protein interactions (PPIs. Förster resonance energy transfer (FRET-based methods have become particularly attractive as they allow quantitative studies of PPIs within the convenient and relevant context of living cells. We describe here an approach that allows the rapid construction of live-cell FRET-based binding curves using a commercially available flow cytometer. We illustrate a simple method for absolutely calibrating the cytometer, validating our binding assay against the gold standard isothermal calorimetry (ITC, and using flow cytometric FRET to uncover the structural and functional effects of the Cushing-syndrome-causing mutation (L206R on PKA’s catalytic subunit. We discover that this mutation not only differentially affects PKAcat’s binding to its multiple partners but also impacts its rate of catalysis. These findings improve our mechanistic understanding of this disease-causing mutation, while illustrating the simplicity, general applicability, and power of flow cytometric FRET.

  16. 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.

  17. 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.

  18. Quantitative multi-color FRET measurements by Fourier lifetime excitation-emission matrix spectroscopy

    Science.gov (United States)

    Zhao, Ming; Huang, Run; Peng, Leilei

    2012-01-01

    Förster resonant energy transfer (FRET) is extensively used to probe macromolecular interactions and conformation changes. The established FRET lifetime analysis method measures the FRET process through its effect on the donor lifetime. In this paper we present a method that directly probes the time-resolved FRET signal with frequency domain Fourier lifetime excitation-emission matrix (FLEEM) measurements. FLEEM separates fluorescent signals by their different phonon energy pathways from excitation to emission. The FRET process generates a unique signal channel that is initiated by donor excitation but ends with acceptor emission. Time-resolved analysis of the FRET EEM channel allows direct measurements on the FRET process, unaffected by free fluorophores that might be present in the sample. Together with time-resolved analysis on non-FRET channels, i.e. donor and acceptor EEM channels, time resolved EEM analysis allows precise quantification of FRET in the presence of free fluorophores. The method is extended to three-color FRET processes, where quantification with traditional methods remains challenging because of the significantly increased complexity in the three-way FRET interactions. We demonstrate the time-resolved EEM analysis method with quantification of three-color FRET in incompletely hybridized triple-labeled DNA oligonucleotides. Quantitative measurements of the three-color FRET process in triple-labeled dsDNA are obtained in the presence of free single-labeled ssDNA and double-labeled dsDNA. The results establish a quantification method for studying multi-color FRET between multiple macromolecules in biochemical equilibrium. PMID:23187535

  19. 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.

  20. 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

  1. 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

  2. rFRET: A comprehensive, Matlab-based program for analyzing intensity-based ratiometric microscopic FRET experiments.

    Science.gov (United States)

    Nagy, Peter; Szabó, Ágnes; Váradi, Tímea; Kovács, Tamás; Batta, Gyula; Szöllősi, János

    2016-04-01

    Fluorescence or Förster resonance energy transfer (FRET) remains one of the most widely used methods for assessing protein clustering and conformation. Although it is a method with solid physical foundations, many applications of FRET fall short of providing quantitative results due to inappropriate calibration and controls. This shortcoming is especially valid for microscopy where currently available tools have limited or no capability at all to display parameter distributions or to perform gating. Since users of multiparameter flow cytometry usually apply these tools, the absence of these features in applications developed for microscopic FRET analysis is a significant limitation. Therefore, we developed a graphical user interface-controlled Matlab application for the evaluation of ratiometric, intensity-based microscopic FRET measurements. The program can calculate all the necessary overspill and spectroscopic correction factors and the FRET efficiency and it displays the results on histograms and dot plots. Gating on plots and mask images can be used to limit the calculation to certain parts of the image. It is an important feature of the program that the calculated parameters can be determined by regression methods, maximum likelihood estimation (MLE) and from summed intensities in addition to pixel-by-pixel evaluation. The confidence interval of calculated parameters can be estimated using parameter simulations if the approximate average number of detected photons is known. The program is not only user-friendly, but it provides rich output, it gives the user freedom to choose from different calculation modes and it gives insight into the reliability and distribution of the calculated parameters. © 2016 International Society for Advancement of Cytometry. © 2016 International Society for Advancement of Cytometry.

  3. 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.

  4. 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

  5. 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.

  6. Single cell FRET analysis for the identification of optimal FRET-pairs in Bacillus subtilis using a prototype MEM-FLIM system.

    Directory of Open Access Journals (Sweden)

    Ruud G J Detert Oude Weme

    Full Text Available Protein-protein interactions can be studied in vitro, e.g. with bacterial or yeast two-hybrid systems or surface plasmon resonance. In contrast to in vitro techniques, in vivo studies of protein-protein interactions allow examination of spatial and temporal behavior of such interactions in their native environment. One approach to study protein-protein interactions in vivo is via Förster Resonance Energy Transfer (FRET. Here, FRET efficiency of selected FRET-pairs was studied at the single cell level using sensitized emission and Frequency Domain-Fluorescence Lifetime Imaging Microscopy (FD-FLIM. For FRET-FLIM, a prototype Modulated Electron-Multiplied FLIM system was used, which is, to the best of our knowledge, the first account of Frequency Domain FLIM to analyze FRET in single bacterial cells. To perform FRET-FLIM, we first determined and benchmarked the best fluorescent protein-pair for FRET in Bacillus subtilis using a novel BglBrick-compatible integration vector. We show that GFP-tagRFP is an excellent donor-acceptor pair for B. subtilis in vivo FRET studies. As a proof of concept, selected donor and acceptor fluorescent proteins were fused using a linker that contained a tobacco etch virus (TEV-protease recognition sequence. Induction of TEV-protease results in loss of FRET efficiency and increase in fluorescence lifetime. The loss of FRET efficiency after TEV induction can be followed in time in single cells via time-lapse microscopy. This work will facilitate future studies of in vivo dynamics of protein complexes in single B. subtilis cells.

  7. 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)

  8. 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.

  9. 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

  10. 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

  11. 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.)

  12. 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)

  13. 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.

  14. 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)

  15. 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.)

  16. 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.

  17. 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.

  18. A communication theoretical analysis of FRET-based mobile ad hoc molecular nanonetworks.

    Science.gov (United States)

    Kuscu, Murat; Akan, Ozgur B

    2014-09-01

    Nanonetworks refer to a group of nanosized machines with very basic operational capabilities communicating to each other in order to accomplish more complex tasks such as in-body drug delivery, or chemical defense. Realizing reliable and high-rate communication between these nanomachines is a fundamental problem for the practicality of these nanonetworks. Recently, we have proposed a molecular communication method based on Förster Resonance Energy Transfer (FRET) which is a nonradiative excited state energy transfer phenomenon observed among fluorescent molecules, i.e., fluorophores. We have modeled the FRET-based communication channel considering the fluorophores as single-molecular immobile nanomachines, and shown its reliability at high rates, and practicality at the current stage of nanotechnology. In this study, for the first time in the literature, we investigate the network of mobile nanomachines communicating through FRET. We introduce two novel mobile molecular nanonetworks: FRET-based mobile molecular sensor/actor nanonetwork (FRET-MSAN) which is a distributed system of mobile fluorophores acting as sensor or actor node; and FRET-based mobile ad hoc molecular nanonetwork (FRET-MAMNET) which consists of fluorophore-based nanotransmitter, nanoreceivers and nanorelays. We model the single message propagation based on birth-death processes with continuous time Markov chains. We evaluate the performance of FRET-MSAN and FRET-MAMNET in terms of successful transmission probability and mean extinction time of the messages, system throughput, channel capacity and achievable communication rates.

  19. 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.

  20. Axisymmetric fretting analysis in coated cylinder

    Indian Academy of Sciences (India)

    The results are compared with 2D models (strip and half-plane) to examine their utility ... ferent values of coefficient of friction and elastic mismatch are illustrated through contour plots of ... fatigue life expectancy of coatings. Depending on the ...

  1. 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.

  2. 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

  3. 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.

  4. 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...

  5. 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.

  6. 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

  7. 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.

  8. 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.

  9. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. 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.

  12. 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.

  13. 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.

  14. 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

  15. 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.

  16. 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

  17. 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.

  18. 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)

  19. 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.

  20. 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

  1. 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

  2. Greater autonomy at work

    NARCIS (Netherlands)

    Houtman, I.L.D.

    2004-01-01

    In the past 10 years, workers in the Netherlands increasingly report more decision-making power in their work. This is important for an economy in recession and where workers face greater work demands. It makes work more interesting, creates a healthier work environment, and provides opportunities

  3. 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.

  4. FLIM-FRET image analysis of tryptophan in prostate cancer cells

    Science.gov (United States)

    Periasamy, Ammasi; Alam, Shagufta R.; Svindrych, Zdenek; Wallrabe, Horst

    2017-07-01

    A region of interest (ROI) based quantitative FLIM-FRET image analysis is developed to quantitate the autofluorescence signals of the essential amino acid tryptophan as a biomarker to investigate the metabolism in prostate cancer cells.

  5. 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.

  6. Methodological considerations for global analysis of cellular FLIM/FRET measurements

    Science.gov (United States)

    Adbul Rahim, Nur Aida; Pelet, Serge; Kamm, Roger D.; So, Peter T. C.

    2012-02-01

    Global algorithms can improve the analysis of fluorescence energy transfer (FRET) measurement based on fluorescence lifetime microscopy. However, global analysis of FRET data is also susceptible to experimental artifacts. This work examines several common artifacts and suggests remedial experimental protocols. Specifically, we examined the accuracy of different methods for instrument response extraction and propose an adaptive method based on the mean lifetime of fluorescent proteins. We further examined the effects of image segmentation and a priori constraints on the accuracy of lifetime extraction. Methods to test the applicability of global analysis on cellular data are proposed and demonstrated. The accuracy of global fitting degrades with lower photon count. By systematically tracking the effect of the minimum photon count on lifetime and FRET prefactors when carrying out global analysis, we demonstrate a correction procedure to recover the correct FRET parameters, allowing us to obtain protein interaction information even in dim cellular regions with photon counts as low as 100 per decay curve.

  7. 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.

  8. 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.

  9. 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...

  10. 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...

  11. 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...

  12. 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

  13. 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.

  14. Study of elastoplastic deformations self-fretting of flat cylinders by mandrelling

    International Nuclear Information System (INIS)

    Caron, Roger

    1974-04-01

    An application of the theory of thick tubes to the special case of flat cylinders which have been self-fretted by mandrelling, is presented. The following materials were used: 1 - a soft steel, XC 18 F, considered to be perfectly elastoplastic; 2 - an alloyed steel, 35 NCD 16, designated consolidable. In the first case, the slip trajectories observed on the polished cylinder surface enabled the plastic deformation region to be defined. It was found, in particular, that the average value of the mean boundary radius at the maximum pressure differs very little from that determined using basic formulas. In the second case, the plastic deformations uniformly affect the internal layers, and privileged trajectories do not exist in this region. On the other hand, the ε θ and ε r expansion curves (from deformation measurements), are continuous from the inner radius to the outer radius; the boundary radius was thus localized from considerations of its correspondence with the ε θ -ε r (shearing deformation) at the elastic limit of the material. This characteristic was determined from measurements made using a test piece provided for this purpose. The radii obtained with this method agree with the theoretical radii over only 4/5 of the total deformation, the uncertainty region being taken into consideration. The maximum value of this parameter was determined in such a way as to obtain a return to a completely elastic rest position. (author) [fr

  15. Greater-confinement disposal

    International Nuclear Information System (INIS)

    Trevorrow, L.E.; Schubert, J.P.

    1989-01-01

    Greater-confinement disposal (GCD) is a general term for low-level waste (LLW) disposal technologies that employ natural and/or engineered barriers and provide a degree of confinement greater than that of shallow-land burial (SLB) but possibly less than that of a geologic repository. Thus GCD is associated with lower risk/hazard ratios than SLB. Although any number of disposal technologies might satisfy the definition of GCD, eight have been selected for consideration in this discussion. These technologies include: (1) earth-covered tumuli, (2) concrete structures, both above and below grade, (3) deep trenches, (4) augered shafts, (5) rock cavities, (6) abandoned mines, (7) high-integrity containers, and (8) hydrofracture. Each of these technologies employ several operations that are mature,however, some are at more advanced stages of development and demonstration than others. Each is defined and further described by information on design, advantages and disadvantages, special equipment requirements, and characteristic operations such as construction, waste emplacement, and closure

  16. More features, greater connectivity.

    Science.gov (United States)

    Hunt, Sarah

    2015-09-01

    Changes in our political infrastructure, the continuing frailties of our economy, and a stark growth in population, have greatly impacted upon the perceived stability of the NHS. Healthcare teams have had to adapt to these changes, and so too have the technologies upon which they rely to deliver first-class patient care. Here Sarah Hunt, marketing co-ordinator at Aid Call, assesses how the changing healthcare environment has affected one of its fundamental technologies - the nurse call system, argues the case for wireless such systems in terms of what the company claims is greater adaptability to changing needs, and considers the ever-wider range of features and functions available from today's nurse call equipment, particularly via connectivity with both mobile devices, and ancillaries ranging from enuresis sensors to staff attack alert 'badges'.

  17. Greater oil investment opportunities

    International Nuclear Information System (INIS)

    Arenas, Ismael Enrique

    1997-01-01

    Geologically speaking, Colombia is a very attractive country for the world oil community. According to this philosophy new and important steps are being taken to reinforce the oil sector: Expansion of the exploratory frontier by including a larger number of sedimentary areas, and the adoption of innovative contracting instruments. Colombia has to offer, Greater economic incentives for the exploration of new areas to expand the exploratory frontier, stimulation of exploration in areas with prospectivity for small fields. Companies may offer Ecopetrol a participation in production over and above royalties, without it's participating in the investments and costs of these fields, more favorable conditions for natural gas seeking projects, in comparison with those governing the terms for oil

  18. 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...

  19. 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

  20. 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

  1. 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.

  2. 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.

  3. Quantitative Analysis of Tau-Microtubule Interaction Using FRET

    Directory of Open Access Journals (Sweden)

    Isabelle L. Di Maïo

    2014-08-01

    Full Text Available The interaction between the microtubule associated protein, tau and the microtubules is investigated. A fluorescence resonance energy transfer (FRET assay was used to determine the distance separating tau to the microtubule wall, as well as the binding parameters of the interaction. By using microtubules stabilized with Flutax-2 as donor and tau labeled with rhodamine as acceptor, a donor-to-acceptor distance of 54 ± 1 Å was found. A molecular model is proposed in which Flutax-2 is directly accessible to tau-rhodamine molecules for energy transfer. By titration, we calculated the stoichiometric dissociation constant to be equal to 1.0 ± 0.5 µM. The influence of the C-terminal tails of αβ-tubulin on the tau-microtubule interaction is presented once a procedure to form homogeneous solution of cleaved tubulin has been determined. The results indicate that the C-terminal tails of α- and β-tubulin by electrostatic effects and of recruitment seem to be involved in the binding mechanism of tau.

  4. 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

  5. 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

  6. 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.

  7. 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.

  8. Experimental facility design for study of fretting in steam generator tubes

    International Nuclear Information System (INIS)

    Balbiani, J.P.; Bergant, M.; Yawny, A.

    2012-01-01

    The design of an experimental facility for fretting wear testing of steam generator tubes under pressurized water up to 340 o C, is presented. The main component of the device consists in an autoclave which permits to recreate steam generator operating conditions. CAD CATIA V5R18, CAE ABAQUS and ASME Sec. VII Div. 1 (Rules for Construction of Pressure Vessels) were used along the design process. The design of the autoclave included the pressure vessel itself and the necessary flanges and nozzles. In addition, an axial dynamic sealing system was designed to allow for actuation from outside the pressure boundary. Complementary, typical tube - support contact conditions were analyzed and the principal variables affecting their mutual interaction determined. In addition, a simple device which allows performing fretting wear testing on steam generator tubes in air at room temperature was fabricated and the feasibility of a quantitative assessment of different aspects related with the fretting induced damage was explored. Characterization techniques available at Centro Atomico Bariloche, like light microscopy, scanning electron microscopy (SEM), energy dispersive analysis of X-ray (EDAX) and surface damage analysis by optic profilometry were shown to be appropriate for this aim. The designed facility will allow evaluating fretting damage of tubes - support combinations that might be used on the steam generator of the prototype reactor CAREM-25. It is also expected it could be applied to characterize fretting severity in other applications (nuclear fuel elements) (author)

  9. 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.

  10. 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.

  11. NIR-Emitting Alloyed CdTeSe QDs and Organic Dye Assemblies: A Nontoxic, Stable, and Efficient FRET System

    Directory of Open Access Journals (Sweden)

    Doris E. Ramírez-Herrera

    2018-04-01

    Full Text Available In the present work, we synthesize Near Infrared (NIR-emitting alloyed mercaptopropionic acid (MPA-capped CdTeSe quantum dots (QDs in a single-step one-hour process, without the use of an inert atmosphere or any pyrophoric ligands. The quantum dots are water soluble, non-toxic, and highly photostable and have high quantum yields (QYs up to 84%. The alloyed MPA-capped CdTeSe QDs exhibit a red-shifted emission, whose color can be tuned between visible and NIR regions (608–750 nm by controlling the Te:Se molar ratio in the precursor mixtures and/or changing the time reaction. The MPA-capped QDs were characterized by UV-visible absorption spectroscopy, fluorescence spectroscopy, transmission electron microscopy (TEM, energy dispersive X-ray spectroscopy (EDS, and zeta potential measurements. Photostability studies were performed by irradiating the QDs with a high-power xenon lamp. The ternary MPA-CdTeSe QDs showed greater photostability than the corresponding binary MPA-CdTe QDs. We report the Förster resonance energy transfer (FRET from the MPA-capped CdTeSe QDs as energy donors and Cyanine5 NHS-ester (Cy5 dye as an energy acceptor with efficiency (E up to 95%. The distance between the QDs and dye (r, the Förster distance (R0, and the binding constant (K are reported. Additionally, cytocompatibility and cell internalization experiments conducted on human cancer cells (HeLa cells revealed that alloyed MPA-capped CdTeSe QDs are more cytocompatible than MPA-capped CdTe QDs and are capable of ordering homogeneously all over the cytoplasm, which allows their use as potential safe, green donors for biological FRET applications.

  12. 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.

  13. 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.

  14. 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...

  15. Fluorophores, environments, and quantification techniques in the analysis of transmembrane helix interaction using FRET.

    Science.gov (United States)

    Khadria, Ambalika S; Senes, Alessandro

    2015-07-01

    Förster resonance energy transfer (FRET) has been widely used as a spectroscopic tool in vitro to study the interactions between transmembrane (TM) helices in detergent and lipid environments. This technique has been instrumental to many studies that have greatly contributed to quantitative understanding of the physical principles that govern helix-helix interactions in the membrane. These studies have also improved our understanding of the biological role of oligomerization in membrane proteins. In this review, we focus on the combinations of fluorophores used, the membrane mimetic environments, and measurement techniques that have been applied to study model systems as well as biological oligomeric complexes in vitro. We highlight the different formalisms used to calculate FRET efficiency and the challenges associated with accurate quantification. The goal is to provide the reader with a comparative summary of the relevant literature for planning and designing FRET experiments aimed at measuring TM helix-helix associations. © 2015 Wiley Periodicals, Inc.

  16. 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)

  17. 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.

  18. 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.

  19. 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.

  20. 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+).

  1. 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...

  2. 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)

  3. 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

  4. 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)

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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.

  12. 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

  13. 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

  14. 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

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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

  1. 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.

  2. 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...

  3. 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

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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

  10. 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.

  11. 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

  12. 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).

  13. 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.)

  14. 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.)

  15. 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.

  16. 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.

  17. 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.

  18. 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)

  19. 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.

  20. 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

  1. 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

  2. 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.

  3. 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.

  4. 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)

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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.

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

    Directory of Open Access Journals (Sweden)

    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.

  13. 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.

  14. Values Research

    Science.gov (United States)

    2009-10-01

    that individualistic employees in individualistic organizations and collectivistic employees in collectivistic organizations show greater job...with Parsons’ causal assumption, in the nineties values were recognized on top of the cultural control –values control norms which in turn control...determines intention which may end in behavior. 7 Defining Human Values Cross- cultural theories on values emerged in the 80s developed by three main

  15. 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.

  16. 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

    An aqueous suspension of fluorescent nanoparticles (PHNNPs) of naphthol based fluorescent organic compound 1-[(Z)-(2-phenylhydrazinylidene) methyl] naphthalene -2-ol (PHN) were prepared using reprecipitation method shows bathochromically shifted aggregation induced enhanced emission (AIEE) in the spectral region where erythrosine (ETS) food dye absorbs strongly. The average size of 72.6 nm of aqueous suspension of PHNNPs obtained by Dynamic light scattering results shows a narrow particle size distribution. The negative zeta potential of nano probe (-22.6 mV) responsible to adsorb oppositely charged analyte on its surface and further permit to bind nano probe and analyte within the close distance proximity required for efficient fluorescence resonance energy transfer (FRET) to take place from donor (PHNNPs) to acceptor (ETS). Systematic FRET experiments performed by measuring fluorescence quenching of PHNNPs with successive addition of ETS solution exploited the use of the PHNNPs as a novel nano probe for the detection of ETS in aqueous solution with extremely lower limit of detection equal to 3.6 nM (3.1 ng/mL). The estimation of photo kinetic and thermodynamic parameters such as quenching rate constant, enthalpy change (∆H), Gibbs free energy change (∆G) and entropy change (∆S) was obtained by the quenching results obtained at different constant temperatures which were found to fit the well-known Stern-Volmer relation. The mechanism of binding and fluorescence quenching of PHNNPs by ETS food dye is proposed on the basis of results obtained in photophysical studies, thermodynamic parameter, energy transfer efficiency, critical energy transfer distance (R0) and distance of approach between donor-acceptor molecules (r). The proposed FRET method based on fluorescence quenching of PHNNPs was successfully applied to develop an analytical method for estimation of ETS from food stuffs without interference of other complex ingredients. Graphical Abstract A

  17. 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

  18. 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

  19. 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

  20. 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

  1. 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.

  2. 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.

  3. A Quantitative Theoretical Framework For Protein-Induced Fluorescence Enhancement-Förster-Type Resonance Energy Transfer (PIFE-FRET).

    Science.gov (United States)

    Lerner, Eitan; Ploetz, Evelyn; Hohlbein, Johannes; Cordes, Thorben; Weiss, Shimon

    2016-07-07

    Single-molecule, protein-induced fluorescence enhancement (PIFE) serves as a molecular ruler at molecular distances inaccessible to other spectroscopic rulers such as Förster-type resonance energy transfer (FRET) or photoinduced electron transfer. In order to provide two simultaneous measurements of two distances on different molecular length scales for the analysis of macromolecular complexes, we and others recently combined measurements of PIFE and FRET (PIFE-FRET) on the single molecule level. PIFE relies on steric hindrance of the fluorophore Cy3, which is covalently attached to a biomolecule of interest, to rotate out of an excited-state trans isomer to the cis isomer through a 90° intermediate. In this work, we provide a theoretical framework that accounts for relevant photophysical and kinetic parameters of PIFE-FRET, show how this framework allows the extraction of the fold-decrease in isomerization mobility from experimental data, and show how these results provide information on changes in the accessible volume of Cy3. The utility of this model is then demonstrated for experimental results on PIFE-FRET measurement of different protein-DNA interactions. The proposed model and extracted parameters could serve as a benchmark to allow quantitative comparison of PIFE effects in different biological systems.

  4. 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.

  5. 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.

  6. 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.

  7. 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

  8. 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

  9. 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)

  10. 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.

  11. 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

  12. 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.

  13. 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.

  14. 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.

  15. 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...

  16. 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...

  17. 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...

  18. [Autoerotic fatalities in Greater Dusseldorf].

    Science.gov (United States)

    Hartung, Benno; Hellen, Florence; Borchard, Nora; Huckenbeck, Wolfgang

    2011-01-01

    Autoerotic fatalities in the Greater Dusseldorf area correspond to the relevant medicolegal literature. Our results included exclusively young to middle-aged, usually single men who were found dead in their city apartments. Clothing and devices used showed a great variety. Women's or fetish clothing and complex shackling or hanging devices were disproportionately frequent. In most cases, death occurred due to hanging or ligature strangulation. There was no increased incidence of underlying psychiatric disorders. In most of the deceased no or at least no remarkable alcohol intoxication was found. Occasionally, it may be difficult to reliably differentiate autoerotic accidents, accidents occurring in connection with practices of bondage & discipline, dominance & submission (BDSM) from natural death, suicide or homicide.

  19. Planning for greater confinement disposal

    International Nuclear Information System (INIS)

    Gilbert, T.L.; Luner, C.; Meshkov, N.K.; Trevorrow, L.E.; Yu, C.

    1985-01-01

    A report that provides guidance for planning for greater-confinement disposal (GCD) of low-level radioactive waste is being prepared. The report addresses procedures for selecting a GCD technology and provides information for implementing these procedures. The focus is on GCD; planning aspects common to GCD and shallow-land burial are covered by reference. Planning procedure topics covered include regulatory requirements, waste characterization, benefit-cost-risk assessment and pathway analysis methodologies, determination of need, waste-acceptance criteria, performance objectives, and comparative assessment of attributes that support these objectives. The major technologies covered include augered shafts, deep trenches, engineered structures, hydrofracture, improved waste forms, and high-integrity containers. Descriptive information is provided, and attributes that are relevant for risk assessment and operational requirements are given. 10 refs., 3 figs., 2 tabs

  20. CdSe/ZnS quantum dots conjugated with a fluorescein derivative: a FRET-based pH sensor for physiological alkaline conditions.

    Science.gov (United States)

    Kurabayashi, Tomokazu; Funaki, Nayuta; Fukuda, Takeshi; Akiyama, Shinnosuke; Suzuki, Miho

    2014-01-01

    Dual pH-dependent fluorescence peaks from a semiconductor quantum dot (QD) and a pH-dependent fluorescent dye can be measured by irradiating with a single wavelength light, and the pH can be estimated from the ratio of the fluorescent intensity of the two peaks. In this work, ratiometric pH sensing was achieved in an aqueous environment by a fluorescent CdSe/ZnS QD appended with a pH-sensitive organic dye, based on fluorescence resonance energy transfer (FRET). By functionalizing the CdSe/ZnS QD with 5-(and 6)-carboxynaphthofluorescein succinimidyl ester as a pH-dependent fluorescent dye, we succeeded in fabricating sensitive nanocomplexes with a linear response to a broad range of physiological pH levels (7.5-9.5) when excited at 450 nm. We found that a purification process is important for increasing the high-fluorescence intensity ratio of a ratiometric fluorescence pH-sensor, and the fluorescence intensity ratio was improved up to 1.0 at pH 8.0 after the purification process to remove unreacted CdSe/ZnS QDs even though the fluorescence of the dye could not be observed without the purification process. The fluorescence intensity ratio corresponds to the fluorescence intensity of the dye, and this fluorescent dye exhibited pH-dependent fluorescence intensity changes. These facts indicate that the fluorescence intensity ratio linearly increased with increasing pH value of the buffer solution containing the QD and the dye. The FRET efficiencies changed from 0.3 (pH 7.5) to 6.2 (pH 9.5).

  1. 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.

  2. [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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. Waste management in Greater Vancouver

    Energy Technology Data Exchange (ETDEWEB)

    Carrusca, K. [Greater Vancouver Regional District, Burnaby, BC (Canada); Richter, R. [Montenay Inc., Vancouver, BC (Canada)]|[Veolia Environmental Services, Vancouver, BC (Canada)

    2006-07-01

    An outline of the Greater Vancouver Regional District (GVRD) waste-to-energy program was presented. The GVRD has an annual budget for solid waste management of $90 million. Energy recovery revenues from solid waste currently exceed $10 million. Over 1,660,00 tonnes of GVRD waste is recycled, and another 280,000 tonnes is converted from waste to energy. The GVRD waste-to-energy facility combines state-of-the-art combustion and air pollution control, and has processed over 5 million tonnes of municipal solid waste since it opened in 1988. Its central location minimizes haul distance, and it was originally sited to utilize steam through sales to a recycle paper mill. The facility has won several awards, including the Solid Waste Association of North America award for best facility in 1990. The facility focuses on continual improvement, and has installed a carbon injection system; an ammonia injection system; a flyash stabilization system; and heat capacity upgrades in addition to conducting continuous waste composition studies. Continuous air emissions monitoring is also conducted at the plant, which produces a very small percentage of the total air emissions in metropolitan Vancouver. The GVRD is now seeking options for the management of a further 500,000 tonnes per year of solid waste, and has received 23 submissions from a range of waste energy technologies which are now being evaluated. It was concluded that waste-to-energy plants can be located in densely populated metropolitan areas and provide a local disposal solution as well as a source of renewable energy. Other GVRD waste reduction policies were also reviewed. refs., tabs., figs.

  13. 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

  14. 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

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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

  1. 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.

  2. 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.

  3. Highly sensitive and quantitative FRET-FLIM imaging in single dendritic spines using improved non-radiative YFP.

    Science.gov (United States)

    Murakoshi, Hideji; Lee, Seok-Jin; Yasuda, Ryohei

    2008-08-01

    Two-photon fluorescence lifetime imaging microscopy (TPFLIM) enables the quantitative measurements of fluorescence resonance energy transfer (FRET) in small subcellular compartments in light scattering tissue. We evaluated and optimized the FRET pair of mEGFP (monomeric EGFP with the A206K mutation) and REACh (non-radiative YFP variants) for TPFLIM. We characterized several mutants of REACh in terms of their "darkness," and their ability to act as a FRET acceptor for mEGFP in HeLa cells and hippocampal neurons. Since the commonly used monomeric mutation A206K increases the brightness of REACh, we introduced a different monomeric mutation (F223R) which does not affect the brightness. Also, we found that the folding efficiency of original REACh, as measured by the fluorescence lifetime of a mEGFP-REACh tandem dimer, was low and variable from cell to cell. Introducing two folding mutations (F46L, Q69M) into REACh increased the folding efficiency by approximately 50%, and reduced the variability of FRET signal. Pairing mEGFP with the new REACh (super-REACh, or sREACh) improved the signal-to-noise ratio compared to the mEGFP-mRFP or mEGFP-original REACh pair by approximately 50%. Using this new pair, we demonstrated that the fraction of actin monomers in filamentous and globular forms in single dendritic spines can be quantitatively measured with high sensitivity. Thus, the mEGFP-sREACh pair is suited for quantitative FRET measurement by TPFLIM, and enables us to measure protein-protein interactions in individual dendritic spines in brain slices with high sensitivity.

  4. 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

  5. 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

  6. 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

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. Optical Fiber Demodulation System with High Performance for Assessing Fretting Damage of Steam Generator Tubes.

    Science.gov (United States)

    Huang, Peijian; Wang, Ning; Li, Junying; Zhu, Yong; Zhang, Jie; Xi, Zhide

    2018-01-12

    In order to access the fretting damage of the steam generator tube (SGT), a fast fiber Fabry-Perot (F-P) non-scanning correlation demodulation system based on a super luminescent light emitting diode (SLED) was performed. By demodulating the light signal coming out from the F-P force sensor, the radial collision force between the SGT and the tube support plate (TSP) was interrogated. For higher demodulation accuracy, the effects of the center wavelength, bandwidth, and spectrum noise of SLED were discussed in detail. Specially, a piezoelectric ceramic transducer (PZT) modulation method was developed to get rid of the interference of mode coupling induced by different types of fiber optics in the demodulation system. The reflectivity of optical wedge and F-P sensor was optimized. Finally, the demodulation system worked well in a 1:1 steam generator test loop and successfully demodulated a force signal of 32 N with a collision time of 2 ms.

  12. 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

  13. 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."

  14. 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

  15. 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.

  16. 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)

  17. 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.

  18. 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.

  19. 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.

  20. 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.

  1. 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

  2. The use of Fluorescence Resonance Energy Transfer (FRET peptidesfor measurement of clinically important proteolytic enzymes

    Directory of Open Access Journals (Sweden)

    Adriana K. Carmona

    2009-09-01

    Full Text Available Proteolytic enzymes have a fundamental role in many biological processes and are associated with multiple pathological conditions. Therefore, targeting these enzymes may be important for a better understanding of their function and development of therapeutic inhibitors. Fluorescence Resonance Energy Transfer (FRET peptides are convenient tools for the study of peptidases specificity as they allow monitoring of the reaction on a continuous basis, providing a rapid method for the determination of enzymatic activity. Hydrolysis of a peptide bond between the donor/acceptor pair generates fluorescence that permits the measurement of the activity of nanomolar concentrations of the enzyme. The assays can be performed directly in a cuvette of the fluorimeter or adapted for determinations in a 96-well fluorescence plate reader. The synthesis of FRET peptides containing ortho-aminobenzoic acid (Abz as fluorescent group and 2, 4-dinitrophenyl (Dnp or N-(2, 4-dinitrophenylethylenediamine (EDDnp as quencher was optimized by our group and became an important line of research at the Department of Biophysics of the Federal University of São Paulo. Recently, Abz/Dnp FRET peptide libraries were developed allowing high-throughput screening of peptidases substrate specificity. This review presents the consolidation of our research activities undertaken between 1993 and 2008 on the synthesis of peptides and study of peptidases specificities.As enzimas proteolíticas têm um papel fundamental em muitos processos biológicos e estão associadas a vários estados patológicos. Por isso, o estudo da especificidade das peptidases pode ser importante para uma melhor compreensão da função destas enzimas e para o desenvolvimento de inibidores. Os substratos com supressão intramolecular de fluorescência constituem uma excelente ferramenta, pois permitem o monitoramento da reação de forma contínua, proporcionando um método prático e rápido para a determinação da

  3. 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.

  4. 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)

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. Materials for coatings against erosion, fretting, and high-temperature oxidation

    International Nuclear Information System (INIS)

    Feller, H.G.; Wienstroth, U.; Balke, C.

    1990-01-01

    This paper investigates the applicability of Co-Cr-W alloys (CoCr29W29, CoCr29W9Y1, CoCr29W9Fe3Y1, CoCr29W9Y1Al1) as coating materials for the substrates MA 6000 and MA 754. Their properties are compared with those of Amperit 410, which is the alloy NiCo23Cr17Al12.5Y0.5. Their isothermal oxidation behaviour at temperatures up to 1000deg C is found to be better for the most part than that of the commercially available Amperit 410. Furthermore, the oxide shows distinctly better adhesion, so that better results concerning resistance to hot-gas corrosion are expected. The fretting behaviour at room temperature is characterized by very low friction factors and a strong resistance to wear. A comparable behaviour is found for resistance to erosive wear. Specimens tested for 500 hours in the pressurised beam device exhibit only minimal changes of mass in the bond MA 600/coating. Single-particle impact tests reveal that exposure of specimens to high temperatures leads to an increase in mean hardness, which is caused by a solidification of the yttrium-containing phase. (orig./MM) [de

  11. 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

  12. Sub-Ensemble Monitoring of DNA Strand Displacement Using Multiparameter Single-Molecule FRET.

    Science.gov (United States)

    Baltierra-Jasso, Laura E; Morten, Michael J; Magennis, Steven W

    2018-03-05

    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 constant of 10 m -1  s -1 . We also followed the displacement from a DNA three-way junction (3WJ) by ssDNA. The presence of three internal mismatched bases in the middle of the invading strand did not prevent displacement from the 3WJ, but reduced the second-order rate constant by about 50 %. We attribute strand exchange in the dsDNA and 3WJ to a zero-toehold pathway from the blunt-ended duplex arms. The single-molecule approach demonstrated here will be useful for studying complex DNA networks. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. 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.

  14. Stabilization of Nucleosomes by Histone Tails and by FACT Revealed by spFRET Microscopy

    Directory of Open Access Journals (Sweden)

    Maria E. Valieva

    2017-01-01

    Full Text Available A correct chromatin structure is important for cell viability and is tightly regulated by numerous factors. Human protein complex FACT (facilitates chromatin transcription is an essential factor involved in chromatin transcription and cancer development. Here FACT-dependent changes in the structure of single nucleosomes were studied with single-particle Förster resonance energy transfer (spFRET microscopy using nucleosomes labeled with a donor-acceptor pair of fluorophores, which were attached to the adjacent gyres of DNA near the contact between H2A-H2B dimers. Human FACT and its version without the C-terminal domain (CTD and the high mobility group (HMG domain of the structure-specific recognition protein 1 (SSRP1 subunit did not change the structure of the nucleosomes, while FACT without the acidic C-terminal domains of the suppressor of Ty 16 (Spt16 and the SSRP1 subunits caused nucleosome aggregation. Proteolytic removal of histone tails significantly disturbed the nucleosome structure, inducing partial unwrapping of nucleosomal DNA. Human FACT reduced DNA unwrapping and stabilized the structure of tailless nucleosomes. CTD and/or HMG domains of SSRP1 are required for this FACT activity. In contrast, previously it has been shown that yeast FACT unfolds (reorganizes nucleosomes using the CTD domain of SSRP1-like Pol I-binding protein 3 subunit (Pob3. Thus, yeast and human FACT complexes likely utilize the same domains for nucleosome reorganization and stabilization, respectively, and these processes are mechanistically similar.

  15. A virus-MIPs fluorescent sensor based on FRET for highly sensitive detection of JEV.

    Science.gov (United States)

    Liang, Caishuang; Wang, Huan; He, Kui; Chen, Chunyan; Chen, Xiaoming; Gong, Hang; Cai, Changqun

    2016-11-01

    Major stumbling blocks in the recognition and detection of virus are the unstable biological recognition element or the complex detection means. Here a fluorescent sensor based on virus-molecular imprinted polymers (virus-MIPs) was designed for specific recognition and highly sensitive detection of Japanese encephalitis virus (JEV). The virus-MIPs were anchored on the surface of silica microspheres modified by fluorescent dye, pyrene-1-carboxaldehyde (PC). The fluorescence intensity of PC can be enhanced by the principle of fluorescence resonance energy transfer (FRET), where virus acted as energy donor and PC acted as energy acceptor. The enhanced fluorescence intensity was proportional to the concentration of virus in the range of 24-960pM, with a limit of detection (LOD, 3σ) of 9.6pM, and the relative standard deviation was 1.99%. In additional, the specificity study confirmed the resultant MIPs has high-selectivity for JEV. This sensor would become a new key for the detection of virus because of its high sensitive, simple operation, high stability and low cost. Copyright © 2016. Published by Elsevier B.V.

  16. 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.

  17. Optical Fiber Demodulation System with High Performance for Assessing Fretting Damage of Steam Generator Tubes

    Directory of Open Access Journals (Sweden)

    Peijian Huang

    2018-01-01

    Full Text Available In order to access the fretting damage of the steam generator tube (SGT, a fast fiber Fabry-Perot (F-P non-scanning correlation demodulation system based on a super luminescent light emitting diode (SLED was performed. By demodulating the light signal coming out from the F-P force sensor, the radial collision force between the SGT and the tube support plate (TSP was interrogated. For higher demodulation accuracy, the effects of the center wavelength, bandwidth, and spectrum noise of SLED were discussed in detail. Specially, a piezoelectric ceramic transducer (PZT modulation method was developed to get rid of the interference of mode coupling induced by different types of fiber optics in the demodulation system. The reflectivity of optical wedge and F-P sensor was optimized. Finally, the demodulation system worked well in a 1:1 steam generator test loop and successfully demodulated a force signal of 32 N with a collision time of 2 ms.

  18. 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.

  19. Greater trochanteric pain syndrome diagnosis and treatment.

    Science.gov (United States)

    Mallow, Michael; Nazarian, Levon N

    2014-05-01

    Lateral hip pain, or greater trochanteric pain syndrome, is a commonly seen condition; in this article, the relevant anatomy, epidemiology, and evaluation strategies of greater trochanteric pain syndrome are reviewed. Specific attention is focused on imaging of this syndrome and treatment techniques, including ultrasound-guided interventions. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Valuing Essays: Essaying Values

    Science.gov (United States)

    Badley, Graham

    2010-01-01

    The essay regularly comes under attack. It is criticised for being rigidly linear rather than flexible and reflective. I first challenge this view by examining reasons why the essay should be valued as an important genre. Secondly, I propose that in using the essay form students and academics necessarily exemplify their own critical values. Essays…

  1. Time-resolved homo-FRET studies of biotin-streptavidin complexes.

    Science.gov (United States)

    Andreoni, Alessandra; Nardo, Luca; Rigler, Rudolf

    2016-09-01

    Förster resonance energy transfer is a mechanism of fluorescence quenching that is notably useful for characterizing properties of biomolecules and/or their interactions. Here we study water-solutions of Biotin-Streptavidin complexes, in which Biotin is labeled with a rigidly-bound fluorophore that can interact by Förster resonance energy transfer with the fluorophores labeling the other, up to three, Biotins of the same complex. The fluorophore, Atto550, is a Rhodamine analogue. We detect the time-resolved fluorescence decay of the fluorophores with an apparatus endowed with single-photon sensitivity and temporal resolution of ~30ps. The decay profiles we observe for samples containing constant Biotin-Atto550 conjugates and varying Streptavidin concentrations are multi-exponential. Each decay component can be associated with the rate of quenching exerted on each donor by each of the acceptors that label the other Biotin molecules, depending on the binding site they occupy. The main features that lead to this result are that (i) the transition dipole moments of the up-to-four Atto550 fluorophores that label the complexes are fixed as to both relative positions and mutual orientations; (ii) the fluorophores are identical and the role of donor in each Biotin-Streptavidin complex is randomly attributed to the one that has absorbed the excitation light (homo-FRET). Obviously the high-temporal resolution of the excitation-detection apparatus is necessary to discriminate among the fluorescence decay components. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. A comparative study on the fretting wear properties of advanced zirconium fuel cladding materials

    International Nuclear Information System (INIS)

    Lee, Young Ho; Kim, Hyung Kyu; Park, Jeong Yong; Kim, Jun Hwan

    2005-06-01

    Fretting wear tests were carried out in room and high temperature water in order to evaluate the wear properties of new zirconium nuclear fuel claddings (K2∼K6) and the commercial claddings (M5, zirlo and zircaloy-4). The objective is to compare the wear resistance of K2∼K6 claddings with that of the commercial ones at the same test condition. After the wear tests, the average wear volume and the maximum wear depth were evaluated and compared at each test condition. As a result, it is difficult to select the most wear-resistant cladding between the K2∼K6 claddings and the commercial ones. This is because the average wear volume and maximum depth of each cladding included between the scattering range of measured results. However, wear resistance of the tested claddings based on the average wear volume and maximum wear depth could be summarized as follows: K5 > zircaloy-4 > (K2,K3) > (K4,M5) > K6 > zirlo at room temperature, zircaloy-4 > K5 > (K3,K4,zirlo) > (K2,K6) > M5 at high temperature and pressure. Therefore, it is concluded that K5 cladding among the tested new zirconium alloys has relatively higher wear-resistance in room and high temperature condition. In order to examine the wear mechanism, it is necessary to systematically study with the consideration of the alloying element effect and test environment. In this report, the wear test procedure and the wear evaluation method are described in detail

  3. 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.

  4. 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...

  5. 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.

  6. Fretting fatigue cracking of a center guide bolt supporting the combustion chamber in a heavy-duty gas turbine engine

    Energy Technology Data Exchange (ETDEWEB)

    Neidel, Andreas; Fischer, Boromir; Gaedicke, Tobias [Siemens AG, Energy Sector, Gasturbinenwerk Berlin (Germany). Werkstoffprueflabor

    2018-04-01

    The slotted center guide bolt of the center guide feature of the lower part of the outer shell of an annular combustion chamber was found fractured in a heavy-duty gas turbine engine used for power generation, after approximately 5.500 operating hours. The incident was a one-off event and not a recurring incident. No similar events were reported from the fleet; hence the failure was not considered a field issue. The metallurgical root cause investigation that was ordered to determine the failure mechanism revealed that the incident center guide bolt failed by fretting fatigue cracking, a high cycle fatigue (HCF) phenomenon.

  7. 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

  8. 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

  9. Simultaneous bilateral isolated greater trochanter fracture

    Directory of Open Access Journals (Sweden)

    Maruti Kambali

    2013-01-01

    Full Text Available A 48-year-old woman sustained simultaneous isolated bilateral greater trochanteric fracture, following a road traffic accident. The patient presented to us 1 month after the injury. She presented with complaints of pain in the left hip and inability to walk. Roentgenograms revealed displaced comminuted bilateral greater trochanter fractures. The fracture of the left greater trochanter was reduced and fixed internally using the tension band wiring technique. The greater trochanter fracture on the right side was asymptomatic and was managed conservatively. The patient regained full range of motion and use of her hips after a postoperative follow-up of 6 months. Isolated fractures of the greater trochanter are unusual injuries. Because of their relative rarity and the unsettled controversy regarding their etiology and pathogenesis, several methods of treatment have been advocated. Furthermore, the reports of this particular type of injury are not plentiful and the average textbook coverage afforded to this entity is limited. In our study we discuss the mechanism of injury and the various treatment options available.

  10. 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.

  11. 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.

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. 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.

  17. 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)

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

    Directory of Open Access Journals (Sweden)

    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.

  19. Greater trochanteric fracture with occult intertrochanteric extension.

    Science.gov (United States)

    Reiter, Michael; O'Brien, Seth D; Bui-Mansfield, Liem T; Alderete, Joseph

    2013-10-01

    Proximal femoral fractures are frequently encountered in the emergency department (ED). Prompt diagnosis is paramount as delay will exacerbate the already poor outcomes associated with these injuries. In cases where radiography is negative but clinical suspicion remains high, magnetic resonance imaging (MRI) is the study of choice as it has the capability to depict fractures which are occult on other imaging modalities. Awareness of a particular subset of proximal femoral fractures, namely greater trochanteric fractures, is vital for both radiologists and clinicians since it has been well documented that they invariably have an intertrochanteric component which may require surgical management. The detection of intertrochanteric or cervical extension of greater trochanteric fractures has been described utilizing MRI but is underestimated with both computed tomography (CT) and bone scan. Therefore, if MRI is unavailable or contraindicated, the diagnosis of an isolated greater trochanteric fracture should be met with caution. The importance of avoiding this potential pitfall is demonstrated in the following case of an elderly woman with hip pain and CT demonstrating an isolated greater trochanteric fracture who subsequently returned to the ED with a displaced intertrochanteric fracture.

  20. Butterfly valves: greater use in power plants

    International Nuclear Information System (INIS)

    McCoy, M.

    1975-01-01

    Improvements in butterfly valves, particularly in the areas of automatic control and leak tightness are described. The use of butterfly valves in nuclear power plants is discussed. These uses include service in component cooling, containment cooling, and containment isolation. The outlook for further improvements and greater uses is examined. (U.S.)

  1. Greater Somalia, the never-ending dream?

    DEFF Research Database (Denmark)

    Zoppi, Marco

    2015-01-01

    This paper provides an historical analysis of the concept of Greater Somalia, the nationalist project that advocates the political union of all Somali-speaking people, including those inhabiting areas in current Djibouti, Ethiopia and Kenya. The Somali territorial unification project of “lost...

  2. A FRET-based high throughput screening assay to identify inhibitors of anthrax protective antigen binding to capillary morphogenesis gene 2 protein.

    Directory of Open Access Journals (Sweden)

    Michael S Rogers

    Full Text Available Anti-angiogenic therapies are effective for the treatment of cancer, a variety of ocular diseases, and have potential benefits in cardiovascular disease, arthritis, and psoriasis. We have previously shown that anthrax protective antigen (PA, a non-pathogenic component of anthrax toxin, is an inhibitor of angiogenesis, apparently as a result of interaction with the cell surface receptors capillary morphogenesis gene 2 (CMG2 protein and tumor endothelial marker 8 (TEM8. Hence, molecules that bind the anthrax toxin receptors may be effective to slow or halt pathological vascular growth. Here we describe development and testing of an effective homogeneous steady-state fluorescence resonance energy transfer (FRET high throughput screening assay designed to identify molecules that inhibit binding of PA to CMG2. Molecules identified in the screen can serve as potential lead compounds for the development of anti-angiogenic and anti-anthrax therapies. The assay to screen for inhibitors of this protein-protein interaction is sensitive and robust, with observed Z' values as high as 0.92. Preliminary screens conducted with a library of known bioactive compounds identified tannic acid and cisplatin as inhibitors of the PA-CMG2 interaction. We have confirmed that tannic acid both binds CMG2 and has anti-endothelial properties. In contrast, cisplatin appears to inhibit PA-CMG2 interaction by binding both PA and CMG2, and observed cisplatin anti-angiogenic effects are not mediated by interaction with CMG2. This work represents the first reported high throughput screening assay targeting CMG2 to identify possible inhibitors of both angiogenesis and anthrax intoxication.

  3. The Value of Value

    DEFF Research Database (Denmark)

    Sørensen, Asger

    parts of business ethics given prominence to especially one term, namely `value'. The question that interests me is the following: What does the articulation of ethics and morality in terms of values mean for ethics and morality as such. Or, to put the question in a more fashionably way: What......As a social scientist of ethics and morality, Luhmann has noticed the ethical wave that has recently swept across the western world, and states that this particular kind of wave seems to have a wavelength of about one hundred years (cf. Luhmann 1989: 9 ff.). Even though the frequency...... and the regularity of such a phenomenon is both hard to verify and, if true, difficult to explain, it seems fair to say that since the Enlightenment, an approaching fin-de-siecle has brought an increased interest in matters concerning morality and ethics.1 The present peak has in public-political discourse and some...

  4. 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.

  5. 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.

  6. 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...

  7. 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

  8. 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

  9. 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.

  10. 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

  11. 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.

  12. 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.

  13. 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.

  14. Utilization of wind energy in greater Hanover

    International Nuclear Information System (INIS)

    Sahling, U.

    1993-01-01

    Since the beginning of the Eighties, the association of communities of Greater Hanover has dealt intensively with energy and ecopolitical questions in the scope of regional planning. Renewable energy sources play a dominant role in this context. This brochure is the third contribution to the subject ''Energy policy and environmental protection''. Experts as well as possibly interested parties are addressed especially. For all 8 contributions contained, separate entries have been recorded in this database. (BWI) [de

  15. Small cities face greater impact from automation

    OpenAIRE

    Frank, Morgan R.; Sun, Lijun; Cebrian, Manuel; Youn, Hyejin; Rahwan, Iyad

    2017-01-01

    The city has proven to be the most successful form of human agglomeration and provides wide employment opportunities for its dwellers. As advances in robotics and artificial intelligence revive concerns about the impact of automation on jobs, a question looms: How will automation affect employment in cities? Here, we provide a comparative picture of the impact of automation across U.S. urban areas. Small cities will undertake greater adjustments, such as worker displacement and job content su...

  16. The Greater Sekhukhune-CAPABILITY outreach project.

    Science.gov (United States)

    Gregersen, Nerine; Lampret, Julie; Lane, Tony; Christianson, Arnold

    2013-07-01

    The Greater Sekhukhune-CAPABILITY Outreach Project was undertaken in a rural district in Limpopo, South Africa, as part of the European Union-funded CAPABILITY programme to investigate approaches for capacity building for the translation of genetic knowledge into care and prevention of congenital disorders. Based on previous experience of a clinical genetic outreach programme in Limpopo, it aimed to initiate a district clinical genetic service in Greater Sekhukhune to gain knowledge and experience to assist in the implementation and development of medical genetic services in South Africa. Implementing the service in Greater Sekhukhune was impeded by a developing staff shortage in the province and pressure on the health service from the existing HIV/AIDS and TB epidemics. This situation underscores the need for health needs assessment for developing services for the care and prevention of congenital disorders in middle- and low-income countries. However, these impediments stimulated the pioneering of innovate ways to offer medical genetic services in these circumstances, including tele-teaching of nurses and doctors, using cellular phones to enhance clinical care and adapting and assessing the clinical utility of a laboratory test, QF-PCR, for use in the local circumstances.

  17. Operational technology for greater confinement disposal

    International Nuclear Information System (INIS)

    Dickman, P.T.; Vollmer, A.T.; Hunter, P.H.

    1984-12-01

    Procedures and methods for the design and operation of a greater confinement disposal facility using large-diameter boreholes are discussed. It is assumed that the facility would be located at an operating low-level waste disposal site and that only a small portion of the wastes received at the site would require greater confinement disposal. The document is organized into sections addressing: facility planning process; facility construction; waste loading and handling; radiological safety planning; operations procedures; and engineering cost studies. While primarily written for low-level waste management site operators and managers, a detailed economic assessment section is included that should assist planners in performing cost analyses. Economic assessments for both commercial and US government greater confinement disposal facilities are included. The estimated disposal costs range from $27 to $104 per cubic foot for a commercial facility and from $17 to $60 per cubic foot for a government facility. These costs are based on average site preparation, construction, and waste loading costs for both contact- and remote-handled wastes. 14 figures, 22 tables

  18. Fast-NPS-A Markov Chain Monte Carlo-based analysis tool to obtain structural information from single-molecule FRET measurements

    Science.gov (United States)

    Eilert, Tobias; Beckers, Maximilian; Drechsler, Florian; Michaelis, Jens

    2017-10-01

    The analysis tool and software package Fast-NPS can be used to analyse smFRET data to obtain quantitative structural information about macromolecules in their natural environment. In the algorithm a Bayesian model gives rise to a multivariate probability distribution describing the uncertainty of the structure determination. Since Fast-NPS aims to be an easy-to-use general-purpose analysis tool for a large variety of smFRET networks, we established an MCMC based sampling engine that approximates the target distribution and requires no parameter specification by the user at all. For an efficient local exploration we automatically adapt the multivariate proposal kernel according to the shape of the target distribution. In order to handle multimodality, the sampler is equipped with a parallel tempering scheme that is fully adaptive with respect to temperature spacing and number of chains. Since the molecular surrounding of a dye molecule affects its spatial mobility and thus the smFRET efficiency, we introduce dye models which can be selected for every dye molecule individually. These models allow the user to represent the smFRET network in great detail leading to an increased localisation precision. Finally, a tool to validate the chosen model combination is provided. Programme Files doi:http://dx.doi.org/10.17632/7ztzj63r68.1 Licencing provisions: Apache-2.0 Programming language: GUI in MATLAB (The MathWorks) and the core sampling engine in C++ Nature of problem: Sampling of highly diverse multivariate probability distributions in order to solve for macromolecular structures from smFRET data. Solution method: MCMC algorithm with fully adaptive proposal kernel and parallel tempering scheme.

  19. ADR characteristics and corporate governance in the Greater China region

    Directory of Open Access Journals (Sweden)

    Lee-Hsien Pan

    2012-04-01

    Full Text Available We examine the relationship between firm valuation and governance mechanisms, firm characteristics, and institutional factors of the American Depository Receipts (ADRs domiciled in the Greater China region. We find that China ADRs have the highest market-to-book value ratio followed by Hong Kong and Taiwan ADRs. It appears that Chinese firms with the poorest external governance environment stand to benefit the most from cross listing under the ADR programs. Listing in the U.S. that requires more stringent regulations and disclosure rules may strengthen the firms’ governance practices and thereby enhance their firm value. Among the internal governance mechanisms, institutional ownership and insider ownership are important for firm value.

  20. Greater happiness for a greater number: Is that possible in Austria?

    NARCIS (Netherlands)

    R. Veenhoven (Ruut)

    2011-01-01

    textabstractWhat is the final goal of public policy? Jeremy Bentham (1789) would say: greater happiness for a greater number. He thought of happiness as subjective enjoyment of life; in his words as “the sum of pleasures and pains”. In his time the happiness of the great number could not be measured

  1. Greater happiness for a greater number: Is that possible? If so how? (Arabic)

    NARCIS (Netherlands)

    R. Veenhoven (Ruut); E. Samuel (Emad)

    2012-01-01

    textabstractWhat is the final goal of public policy? Jeremy Bentham (1789) would say: greater happiness for a greater number. He thought of happiness as subjective enjoyment of life; in his words as “the sum of pleasures and pains”. In his time, the happiness of the great number could not be

  2. Greater happiness for a greater number: Is that possible in Germany?

    NARCIS (Netherlands)

    R. Veenhoven (Ruut)

    2009-01-01

    textabstractWhat is the final goal of public policy? Jeremy Bentham (1789) would say: greater happiness for a greater number. He thought of happiness as subjective enjoyment of life; in his words as “the sum of pleasures and pains”. In his time the Happiness of the great number could not be measured

  3. Broadband Light-Harvesting Molecular Triads with High FRET Efficiency Based on the Coumarin-Rhodamine-BODIPY Platform.

    Science.gov (United States)

    He, Longwei; Zhu, Sasa; Liu, Yong; Xie, Yinan; Xu, Qiuyan; Wei, Haipeng; Lin, Weiying

    2015-08-17

    Broadband capturing and FRET-based light-harvesting molecular triads, CRBs, based on the coumarin-rhodamine-BODIPY platform were rationally designed and synthesized. The absorption band of CRBs starts from blue-green to yellow-orange regions (330-610 nm), covering the strong radiation scope of sunlight. The peripheral coumarin and BODIPY chromophore energy could transfer to the central acceptor rhodamine by a one-step direct way. The energy of the coumarin moiety could also transfer to the BODIPY unit, subsequently transferring to the rhodamine core by two-step sequential ways. Both the efficiencies of the coumarin moiety and the BODIPY unit to the rhodamine core in CRBs, determined by two different ways, are very high. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. 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

  5. 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

  6. 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.

  7. Ratiometric FRET-based detection of DNA and micro-RNA on the surface using TIRF detection

    International Nuclear Information System (INIS)

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

    2010-01-01

    A new FRET-based method for the ratiometric detection of DNA oligomers on a surface using TIRF detection mode is reported. The dual-labeled system consisting of two hybridized oligomers, Cy3oligoY:Cy5oligoX was immobilized on the surface, and the total internal reflection fluorescence (TIRF) was used to detect emission signals from the surface. Two signals, green and red, which originated from the green donor Cy3 and the red acceptor Cy5, have been simultaneously detected. When the target single-stranded complimentary oligomer was present in the solution, this oligomer replaced the Cy3oligoY in the donor:acceptor complex on the surface and the ratio of red-to-green signal was dramatically changed. This detection scheme is generally applicable to the detection of DNA or RNA on a surface.

  8. A Series of Fluorescent and Colorimetric Chemodosimeters for Selective Recognition of Cyanide Based on the FRET Mechanism.

    Science.gov (United States)

    Hua, Ying-Xi; Shao, Yongliang; Wang, Ya-Wen; Peng, Yu

    2017-06-16

    A series of fluorescence "turn-on" probes (PY, AN, NA, B1, and B2) have been developed and successfully applied to detect cyanide anions based on the Michael addition reaction and FRET mechanism. These probes demonstrated good selectivity, high sensitivity, and very fast recognition for CN - . In particular, the fluorescence response of probe NA finished within 3 s. Low limits of detection (down to 63 nM) are also obtained in these probes with remarkable fluorescence enhancement factors. In addition, fluorescence colors of these probes turned to blue, yellow, or orange upon sensing CN - . In UV-vis mode, all of them showed ratiometric response for CN - . 1 H NMR titration experiments and TDDFT calculations were taken to verify the mechanism of the specific reaction and fluorescence properties of the corresponding compounds. Moreover, silica gel plates with these probes were also fabricated and utilized to detect cyanide.

  9. Safety assessment of the potential for foreign object - caused fretting - wear damages on PWR steam generator U-tubes

    International Nuclear Information System (INIS)

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

    2003-01-01

    This study investigates the safety assessment of the potential for fretting-wear damages on Steam Generator (SG) U-tubes caused by foreign object in operating nuclear power plants. The operating SG shell-side flow field conditions are obtained from three-dimensional SG flow calculation using the ATHOS3 code. Modal analyses are performed for the finite element modelings of U-tubes to get the natural frequency, corresponding mode shape and participation factor. The wear rate of U-tube caused by foreign object is calculated using the Archard formula and the remaining life of the tube is predicted. Also, discussed in this study are the effects of flow velocity, internal pressure, tube-to-foreign object contact angle, and vibration of the tube on the remaining life of the tube

  10. Life Estimation of PWR Steam Generator U-Tubes Subjected to Foreign Object-Induced Fretting Wear

    International Nuclear Information System (INIS)

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

    2005-01-01

    This paper presents an approach to the remaining life prediction of steam generator (SG) U-tubes, which are intact initially, subjected to fretting-wear degradation due to the interaction between a vibrating tube and a foreign object in operating nuclear power plants. The operating SG shell-side flow field conditions are obtained from a three-dimensional SG flow calculation using the ATHOS3 code. Modal analyses are performed for the finite element models of U-tubes to get the natural frequency, corresponding mode shape, and participation factor. The wear rate of a U-tube caused by a foreign object is calculated using the Archard formula, and the remaining life of the tube is predicted. Also discussed in this study are the effects of the tube modal characteristics, external flow velocity, and tube internal pressure on the estimated results of the remaining life of the tube

  11. Search for greater stability in nuclear regulation

    International Nuclear Information System (INIS)

    Asselstine, J.K.

    1985-01-01

    The need for greater stability in nuclear regulation is discussed. Two possible approaches for dealing with the problems of new and rapidly changing regulatory requirements are discussed. The first approach relies on the more traditional licensing reform initiatives that have been considered off and on for the past decade. The second approach considers a new regulator philosophy aimed at the root causes of the proliferation of new safety requirements that have been imposed in recent years. For the past few years, the concepts of deregulation and regulatory reform have been in fashion in Washington, and the commercial nuclear power program has not remained unaffected. Many look to these concepts to provide greater stability in the regulatory program. The NRC, the nuclear industry and the administration have all been avidly pursuing regulatory reform initiatives, which take the form of both legislative and administrative proposals. Many of these proposals look to the future, and, if adopted, would have little impact on currently operating nuclear power plants or plants now under construction

  12. Greater Sudbury fuel efficient driving handbook

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2009-12-15

    Reducing the amount of fuel that people use for personal driving saves money, improves local air quality, and reduces personal contributions to climate change. This handbook was developed to be used as a tool for a fuel efficient driving pilot program in Greater Sudbury in 2009-2010. Specifically, the purpose of the handbook was to provide greater Sudbury drivers with information on how to drive and maintain their personal vehicles in order to maximize fuel efficiency. The handbook also provides tips for purchasing fuel efficient vehicles. It outlines the benefits of fuel maximization, with particular reference to reducing contributions to climate change; reducing emissions of air pollutants; safe driving; and money savings. Some tips for efficient driving are to avoid aggressive driving; use cruise control; plan trips; and remove excess weight. Tips for efficient winter driving are to avoid idling to warm up the engine; use a block heater; remove snow and ice; use snow tires; and check tire pressure. The importance of car maintenance and tire pressure was emphasized. The handbook also explains how fuel consumption ratings are developed by vehicle manufacturers. refs., figs.

  13. Women at greater risk of HIV infection.

    Science.gov (United States)

    Mahathir, M

    1997-04-01

    Although many people believe that mainly men get infected with HIV/AIDS, women are actually getting infected at a faster rate than men, especially in developing countries, and suffer more from the adverse impact of AIDS. As of mid-1996, the Joint UN Program on AIDS estimated that more than 10 million of the 25 million adults infected with HIV since the beginning of the epidemic are women. The proportion of HIV-positive women is growing, with almost half of the 7500 new infections daily occurring among women. 90% of HIV-positive women live in a developing country. In Asia-Pacific, 1.4 million women have been infected with HIV out of an estimated total 3.08 million adults from the late 1970s until late 1994. Biologically, women are more vulnerable than men to infection because of the greater mucus area exposed to HIV during penile penetration. Women under age 17 years are at even greater risk because they have an underdeveloped cervix and low vaginal mucus production. Concurrent sexually transmitted diseases increase the risk of HIV transmission. Women's risk is also related to their exposure to gender inequalities in society. The social and economic pressures of poverty exacerbate women's risk. Prevention programs are discussed.

  14. 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.

  15. 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)

  16. Small cities face greater impact from automation.

    Science.gov (United States)

    Frank, Morgan R; Sun, Lijun; Cebrian, Manuel; Youn, Hyejin; Rahwan, Iyad

    2018-02-01

    The city has proved to be the most successful form of human agglomeration and provides wide employment opportunities for its dwellers. As advances in robotics and artificial intelligence revive concerns about the impact of automation on jobs, a question looms: how will automation affect employment in cities? Here, we provide a comparative picture of the impact of automation across US urban areas. Small cities will undertake greater adjustments, such as worker displacement and job content substitutions. We demonstrate that large cities exhibit increased occupational and skill specialization due to increased abundance of managerial and technical professions. These occupations are not easily automatable, and, thus, reduce the potential impact of automation in large cities. Our results pass several robustness checks including potential errors in the estimation of occupational automation and subsampling of occupations. Our study provides the first empirical law connecting two societal forces: urban agglomeration and automation's impact on employment. © 2018 The Authors.

  17. Small cities face greater impact from automation

    Science.gov (United States)

    Sun, Lijun; Cebrian, Manuel; Rahwan, Iyad

    2018-01-01

    The city has proved to be the most successful form of human agglomeration and provides wide employment opportunities for its dwellers. As advances in robotics and artificial intelligence revive concerns about the impact of automation on jobs, a question looms: how will automation affect employment in cities? Here, we provide a comparative picture of the impact of automation across US urban areas. Small cities will undertake greater adjustments, such as worker displacement and job content substitutions. We demonstrate that large cities exhibit increased occupational and skill specialization due to increased abundance of managerial and technical professions. These occupations are not easily automatable, and, thus, reduce the potential impact of automation in large cities. Our results pass several robustness checks including potential errors in the estimation of occupational automation and subsampling of occupations. Our study provides the first empirical law connecting two societal forces: urban agglomeration and automation's impact on employment. PMID:29436514

  18. Planning for greater-confinement disposal

    International Nuclear Information System (INIS)

    Gilbert, T.L.; Luner, C.; Meshkov, N.K.; Trevorrow, L.E.; Yu, C.

    1984-01-01

    This contribution is a progress report for preparation of a document that will summarize procedures and technical information needed to plan for and implement greater-confinement disposal (GCD) of low-level radioactive waste. Selection of a site and a facility design (Phase I), and construction, operation, and extended care (Phase II) will be covered in the document. This progress report is limited to Phase I. Phase I includes determination of the need for GCD, design alternatives, and selection of a site and facility design. Alternative designs considered are augered shafts, deep trenches, engineered structures, high-integrity containers, hydrofracture, and improved waste form. Design considerations and specifications, performance elements, cost elements, and comparative advantages and disadvantages of the different designs are covered. Procedures are discussed for establishing overall performance objectives and waste-acceptance criteria, and for comparative assessment of the performance and cost of the different alternatives. 16 references

  19. Greater confinement disposal of radioactive wastes

    International Nuclear Information System (INIS)

    Trevorrow, L.E.; Gilbert, T.L.; Luner, C.; Merry-Libby, P.A.; Meshkov, N.K.; Yu, C.

    1985-01-01

    Low-level radioactive waste (LLW) includes a broad spectrum of different radionuclide concentrations, half-lives, and hazards. Standard shallow-land burial practice can provide adequate protection of public health and safety for most LLW. A small volume fraction (approx. 1%) containing most of the activity inventory (approx. 90%) requires specific measures known as greater-confinement disposal (GCD). Different site characteristics and different waste characteristics - such as high radionuclide concentrations, long radionuclide half-lives, high radionuclide mobility, and physical or chemical characteristics that present exceptional hazards - lead to different GCD facility design requirements. Facility design alternatives considered for GCD include the augered shaft, deep trench, engineered structure, hydrofracture, improved waste form, and high-integrity container. Selection of an appropriate design must also consider the interplay between basic risk limits for protection of public health and safety, performance characteristics and objectives, costs, waste-acceptance criteria, waste characteristics, and site characteristics

  20. Planning for greater-confinement disposal

    International Nuclear Information System (INIS)

    Gilbert, T.L.; Luner, C.; Meshkov, N.K.; Trevorrow, L.E.; Yu, C.

    1984-01-01

    This contribution is a progress report for preparation of a document that will summarize procedures and technical information needed to plan for and implement greater-confinement disposal (GCD) of low-level radioactive waste. Selection of a site and a facility design (Phase I), and construction, operation, and extended care (Phase II) will be covered in the document. This progress report is limited to Phase I. Phase I includes determination of the need for GCD, design alternatives, and selection of a site and facility design. Alternative designs considered are augered shafts, deep trenches, engineered structures, high-integrity containers, hydrofracture, and improved waste form. Design considerations and specifications, performance elements, cost elements, and comparative advantages and disadvantages of the different designs are covered. Procedures are discussed for establishing overall performance objecties and waste-acceptance criteria, and for comparative assessment of the performance and cost of the different alternatives. 16 refs

  1. Urban acid deposition in Greater Manchester

    Energy Technology Data Exchange (ETDEWEB)

    Lee, D.S.; Longhurst, J.W.S.; Gee, D.R.; Hare, S.E. (Manchester Polytechnic, Manchester (UK). Acid Rain Information Centre)

    1989-08-01

    Data are presented from a monitoring network of 18 bulk precipitation collectors and one wet-only collector in the urban area of Greater Manchester, in the north west of England. Weekly samples were analysed for all the major ions in precipitation along with gaseous nitrogen dioxide concentrations from diffusion tubes. Statistical analysis of the data shows significant spatial variation of non marine sulphate, nitrate, ammonium, acidity and calcium concentrations, and nitrogen dioxide concentrations. Calcium is thought to be responsible for the buffering of acidity and is of local origin. Wet deposition is the likely removal process for calcium in the atmosphere and probably by below cloud scavenging. Nitrate and ammonium concentrations and depositions show close spatial, temporal and statistical association. Examination of high simultaneous episodes of nitrate and ammonium deposition shows that these depositions cannot be explained in terms of trajectories and it is suggested that UK emissions of ammonia may be important. Statistical analysis of the relationships between nitrate and ammonium depositions, concentrations and precipitation amount suggest that ammonia from mesoscale sources reacts reversibly with nitric acid aerosol and is removed by below cloud scavenging. High episodes of the deposition of non marine sulphate are difficult to explain by trajectory analysis alone, perhaps suggesting local sources. In a comparison between wet deposition and bulk deposition, it was shown that only 15.2% of the non marine sulphur was dry deposited to the bulk precipitation collector. 63 refs., 86 figs., 31 tabs.

  2. 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.

  3. 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

  4. Land use scenarios for greater Copenhagen

    DEFF Research Database (Denmark)

    Fertner, Christian; Jørgensen, Gertrud; Nielsen, Thomas Alexander Sick

    2012-01-01

    Urban planning and development in Denmark can be characterised by a relatively strong planning framework. Land use scenarios based on empirically derived dynamics of urban growth are practically never applied. However, modelling approaches do offer a methodology to explore the pressures in an urban...... region, as well as an approach to understand urban development patterns outside the ‘spatial masterplan’. In this context we will present the results of a modelling exercise addressing future land use change in the metropolitan area of Copenhagen, Denmark, and the impact of the current regional planning...... short period of time. The set-up and the results were discussed with a few experts from the Danish Ministry of the Environment and its value as discussion input recognized. The approach offers a lot of possibilities to discuss urban growth and spatial planning policies, even in a country with a strong...

  5. Public Values

    DEFF Research Database (Denmark)

    Beck Jørgensen, Torben; Rutgers, Mark R.

    2015-01-01

    administration is approached in terms of processes guided or restricted by public values and as public value creating: public management and public policy-making are both concerned with establishing, following and realizing public values. To study public values a broad perspective is needed. The article suggest......This article provides the introduction to a symposium on contemporary public values research. It is argued that the contribution to this symposium represent a Public Values Perspective, distinct from other specific lines of research that also use public value as a core concept. Public...... a research agenda for this encompasing kind of public values research. Finally the contributions to the symposium are introduced....

  6. 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.

  7. 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.

  8. 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.

  9. 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.

  10. 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.

  11. Study on torsional fretting wear behavior of a ball-on-socket contact configuration simulating an artificial cervical disk

    International Nuclear Information System (INIS)

    Wang, Song; Wang, Fei; Liao, Zhenhua; Wang, Qingliang; Liu, Yuhong; Liu, Weiqiang

    2015-01-01

    A ball-on-socket contact configuration was designed to simulate an artificial cervical disk in structure. UHMWPE (ultra high molecular weight polyethylene) hot pressed by powders and Ti6Al4V alloy were selected as the material combination of ball and socket. The socket surface was coated by a ~ 500 nm C-DLC (carbon ion implantation-diamond like carbon) mixed layer to improve its surface nano hardness and wear resistance. The torsional fretting wear behavior of the ball-on-socket model was tested at different angular displacements under 25% bovine serum lubrication with an axial force of 100 N to obtain more realistic results with that in vivo. The fretting running regimes and wear damage characteristics as well as wear mechanisms for both ball and socket were studied based on 2D (two dimension) optical microscope, SEM (scanning electron microscope) and 3D (three dimension) profiles. With the increase of angular displacement amplitude from 1° to 7°, three types of T-θ (Torsional torque-angular displacement amplitude) curves (i.e., linear, elliptical and parallelogram loops) corresponding to running regimes of PSR (partial slip regime), MR (mixed regime) and SR (slip regime) were observed and analyzed. Both the central region and the edge zone of the ball and socket were damaged. The worn surfaces were characterized by wear scratches and wear debris. In addition, more severe wear damage and more wear debris appeared on the central region of the socket at higher angular displacement amplitude. The dominant damage mechanism was a mix of surface scratch, adhesive wear and abrasive wear for the UHMWPE ball while that for the coated socket was abrasive wear by PE particles and some polishing and rolling process on the raised overgrown DLC grains. The frictional kinetic behavior, wear type, damage region and damage mechanism for the ball-on-socket model revealed significant differences with those of a ball-on-flat contact while showing better consistency with that of in

  12. Study on torsional fretting wear behavior of a ball-on-socket contact configuration simulating an artificial cervical disk

    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); Wang, Fei [School of Material Science and Engineering, China University of Mining and Technology, Xuzhou 221116 (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); Wang, Qingliang [School of Material Science and Engineering, China University of Mining and Technology, Xuzhou 221116 (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 [Biomechanics and Biotechnology Lab, Research Institute of Tsinghua University in Shenzhen, Shenzhen 518057 (China); State Key Laboratory of Tribology, Tsinghua University, Beijing 100084 (China)

    2015-10-01

    A ball-on-socket contact configuration was designed to simulate an artificial cervical disk in structure. UHMWPE (ultra high molecular weight polyethylene) hot pressed by powders and Ti6Al4V alloy were selected as the material combination of ball and socket. The socket surface was coated by a ~ 500 nm C-DLC (carbon ion implantation-diamond like carbon) mixed layer to improve its surface nano hardness and wear resistance. The torsional fretting wear behavior of the ball-on-socket model was tested at different angular displacements under 25% bovine serum lubrication with an axial force of 100 N to obtain more realistic results with that in vivo. The fretting running regimes and wear damage characteristics as well as wear mechanisms for both ball and socket were studied based on 2D (two dimension) optical microscope, SEM (scanning electron microscope) and 3D (three dimension) profiles. With the increase of angular displacement amplitude from 1° to 7°, three types of T-θ (Torsional torque-angular displacement amplitude) curves (i.e., linear, elliptical and parallelogram loops) corresponding to running regimes of PSR (partial slip regime), MR (mixed regime) and SR (slip regime) were observed and analyzed. Both the central region and the edge zone of the ball and socket were damaged. The worn surfaces were characterized by wear scratches and wear debris. In addition, more severe wear damage and more wear debris appeared on the central region of the socket at higher angular displacement amplitude. The dominant damage mechanism was a mix of surface scratch, adhesive wear and abrasive wear for the UHMWPE ball while that for the coated socket was abrasive wear by PE particles and some polishing and rolling process on the raised overgrown DLC grains. The frictional kinetic behavior, wear type, damage region and damage mechanism for the ball-on-socket model revealed significant differences with those of a ball-on-flat contact while showing better consistency with that of in

  13. Comparability of reference values

    International Nuclear Information System (INIS)

    Rossbach, M.; Stoeppler, M.

    1993-01-01

    Harmonization of certified values in Reference Materials (RMs) can be carried out by applying nuclear analytical techniques to RMs of various matrix types and concentration levels. Although RMs generally should not be used as primary standards the cross evaluation of concentrations in RMs leads to better compatibility of reference values and thus to a greater agreement between analytical results from different laboratories using these RMs for instrument calibration and quality assurance. (orig.)

  14. 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.

  15. 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

  16. A RhoA-FRET Biosensor Mouse for Intravital Imaging in Normal Tissue Homeostasis and Disease Contexts

    Directory of Open Access Journals (Sweden)

    Max Nobis

    2017-10-01

    Full Text Available The small GTPase RhoA is involved in a variety of fundamental processes in normal tissue. Spatiotemporal control of RhoA is thought to govern mechanosensing, growth, and motility of cells, while its deregulation is associated with disease development. Here, we describe the generation of a RhoA-fluorescence resonance energy transfer (FRET biosensor mouse and its utility for monitoring real-time activity of RhoA in a variety of native tissues in vivo. We assess changes in RhoA activity during mechanosensing of osteocytes within the bone and during neutrophil migration. We also demonstrate spatiotemporal order of RhoA activity within crypt cells of the small intestine and during different stages of mammary gestation. Subsequently, we reveal co-option of RhoA activity in both invasive breast and pancreatic cancers, and we assess drug targeting in these disease settings, illustrating the potential for utilizing this mouse to study RhoA activity in vivo in real time.

  17. Assembly and dynamics of the U4/U6 di-snRNP by single-molecule FRET

    Science.gov (United States)

    Hardin, John W.; Warnasooriya, Chandani; Kondo, Yasushi; Nagai, Kiyoshi; Rueda, David

    2015-01-01

    In large ribonucleoprotein machines, such as ribosomes and spliceosomes, RNA functions as an assembly scaffold as well as a critical catalytic component. Protein binding to the RNA scaffold can induce structural changes, which in turn modulate subsequent binding of other components. The spliceosomal U4/U6 di-snRNP contains extensively base paired U4 and U6 snRNAs, Snu13, Prp31, Prp3 and Prp4, seven Sm and seven LSm proteins. We have studied successive binding of all protein components to the snRNA duplex during di-snRNP assembly by electrophoretic mobility shift assay and accompanying conformational changes in the U4/U6 RNA 3-way junction by single-molecule FRET. Stems I and II of the duplex were found to co-axially stack in free RNA and function as a rigid scaffold during the entire assembly, but the U4 snRNA 5′ stem-loop adopts alternative orientations each stabilized by Prp31 and Prp3/4 binding accounting for altered Prp3/4 binding affinities in presence of Prp31. PMID:26503251

  18. Tight coupling of astrocyte energy metabolism to synaptic activity revealed by genetically encoded FRET nanosensors in hippocampal tissue.

    Science.gov (United States)

    Ruminot, Iván; Schmälzle, Jana; Leyton, Belén; Barros, L Felipe; Deitmer, Joachim W

    2017-01-01

    The potassium ion, K + , a neuronal signal that is released during excitatory synaptic activity, produces acute activation of glucose consumption in cultured astrocytes, a phenomenon mediated by the sodium bicarbonate cotransporter NBCe1 ( SLC4A4). We have explored here the relevance of this mechanism in brain tissue by imaging the effect of neuronal activity on pH, glucose, pyruvate and lactate dynamics in hippocampal astrocytes using BCECF and FRET nanosensors. Electrical stimulation of Schaffer collaterals produced fast activation of glucose consumption in astrocytes with a parallel increase in intracellular pyruvate and biphasic changes in lactate . These responses were blocked by TTX and were absent in tissue slices prepared from NBCe1-KO mice. Direct depolarization of astrocytes with elevated extracellular K + or Ba 2+ mimicked the metabolic effects of electrical stimulation. We conclude that the glycolytic pathway of astrocytes in situ is acutely sensitive to neuronal activity, and that extracellular K + and the NBCe1 cotransporter are involved in metabolic crosstalk between neurons and astrocytes. Glycolytic activation of astrocytes in response to neuronal K + helps to provide an adequate supply of lactate, a metabolite that is released by astrocytes and which acts as neuronal fuel and an intercellular signal.

  19. Characterisation of the surface topography, tomography and chemistry of fretting corrosion product found on retrieved polished femoral stems.

    Science.gov (United States)

    Bryant, M; Ward, M; Farrar, R; Freeman, R; Brummitt, K; Nolan, J; Neville, A

    2014-04-01

    This study presents the characterisation of the surface topography, tomography and chemistry of fretting corrosion product found on retrieved polished femoral stems. Scanning Electron Microscopy (SEM), X-ray Photoelectron Spectroscopy (XPS), Transmission Electron Microscopy (TEM) and Fourier Transform Infrared Spectroscopy (FI-IR) were utilised in order to assess the surface morphology of retrieved Metal-on-Metal Total Hip Replacements and surface chemistry of the films found on the surface. Gross slip, plastic deformation and directionality of the surface were extensively seen on the proximal surfaces of the retrievals. A more corrosive phenomenon was observed in the distal regions of the stem, demonstrating a seemingly intergranular attack. Tribochemical reactions were seen to occur within the stem-cement interfaces with tribofilms being observed on the femoral stem and counterpart PMMA bone cement. XPS, TEM-EDX and FT-IR analyses demonstrated that the films present in the stem surfaces were a complex mixture of chromium oxide and amorphous organic material. A comparison between current experimental and clinical literature has been conducted and findings from this study demonstrate that the formation and chemistry of films are drastically influenced by the type of wear or degradation mechanism. Films formed in the stem-cement interface are thought to further influence the biological environment outside the stem-cement interface due to the formation of Cr and O rich films within the interface whilst Co is free to migrate away. © 2013 Elsevier Ltd. All rights reserved.

  20. Selective Killing of Breast Cancer Cells by Doxorubicin-Loaded Fluorescent Gold Nanoclusters: Confocal Microscopy and FRET.

    Science.gov (United States)

    Chattoraj, Shyamtanu; Amin, Asif; Jana, Batakrishna; Mohapatra, Saswat; Ghosh, Surajit; Bhattacharyya, Kankan

    2016-01-18

    Fluorescent gold nanoclusters (AuNCs) capped with lysozymes are used to deliver the anticancer drug doxorubicin to cancer and noncancer cells. Doxorubicin-loaded AuNCs cause the highly selective and efficient killing (90 %) of breast cancer cells (MCF7) (IC50 =155 nm). In contrast, the killing of the noncancer breast cells (MCF10A) by doxorubicin-loaded AuNCs is only 40 % (IC50 =4500 nm). By using a confocal microscope, the fluorescence spectrum and decay of the AuNCs were recorded inside the cell. The fluorescence maxima (at ≈490-515 nm) and lifetime (≈2 ns), of the AuNCs inside the cells correspond to Au10-13 . The intracellular release of doxorubicin from AuNCs is monitored by Förster resonance energy transfer (FRET) imaging. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Greater Confinement Disposal Program at the Savannah River Plant

    International Nuclear Information System (INIS)

    Towler, O.A.; Cook, J.R.; Peterson, D.L.

    1983-01-01

    Plans for improved LLW disposal at the Savannah River Plant include Greater Confinement Disposal (GCD) for the higher activity fractions of this waste. GCD practices will include waste segregation, packaging, emplacement below the root zone, and stabilizing the emplacement with cement. Statistical review of SRP burial records showed that about 95% of the radioactivity is associated with only 5% of the waste volume. Trigger values determined in this study were compared with actual burials in 1982 to determine what GCD facilities would be needed for a demonstration to begin in Fall 1983. Facilities selected include 8-feet-diameter x 30-feet-deep boreholes to contain reactor scrap, tritiated waste, and selected wastes from offsite

  2. Use of renewable energy in the greater metropolitan area

    International Nuclear Information System (INIS)

    Arias Garcia, Rocio; Castro Gomez, Gustavo; Fallas Cordero, Kenneth; Grant Chaves, Samuel; Mendez Parrales, Tony; Parajeles Fernandez, Ivan

    2012-01-01

    A study is conducted on different renewable energy within the larger metropolitan area, selecting the most suitable for the area and the implementation for distributed generation. A research methodology is practiced type pretending gather the necessary information to make proposals selected of different type of energy. The geography of the greater metropolitan area is studied along with the different existing renewable energy: distributed generation, remote measurement of energy which is one of the elements of the concept of intelligent networks (Smart Grid) in the electricity sector, legislation of Costa Rica regarding the generation of renewable energy and environmental impact. An analysis of economic feasibility is covered for each of the proposals estimating current rates for leading distributors of a future value, concluding with the viability of projects for possible execution of the same. (author) [es

  3. Age and Expatriate Job Performance in Greater China

    DEFF Research Database (Denmark)

    Selmer, Jan; Lauring, Jakob; Feng, Yunxia

    2009-01-01

    a positive impact on expatriates' job performance. Therefore, the purpose of this paper is toexamine the association between the age of business expatriates and their work performance in a Chinese cultural setting. Design/methodology/approach - Controlling for the potential bias of a number of background......, companies should not discriminate against older candidatesin expatriate selection for Greater China. Furthermore, older expatriates destined for a Chinesecultural context could be trained how to exploit their age advantage. Originality/value - In contrast to previous studies, this investigation attempts...... to match a certain personal characteristic of expatriates with a specific host culture. The results have implications for and contribute to the literature on expatriate selection as well as to the body of research on crosscultural training....

  4. Married Women, Work, and Values.

    Science.gov (United States)

    Jalilvand, Mahshid

    2000-01-01

    Working women appear to have a personal-value structure different from that of nonworking women. Economic and political values are more prominent among women who work, whereas social and religious values play a greater role for women who stay at home. (JOW)

  5. The Value of Architecture

    DEFF Research Database (Denmark)

    Beim, Anne

    2018-01-01

    of Europe’s architectural heritage. This resulted in a greater emphasis on building conservation and led to the development of the Survey of Architectural Values in the Environment (SAVE) by the Danish heritage authorities. The value assessment was initially meant for evaluating building heritage...... and authorities in efforts to evaluate the cultural value of the built environment. The National Trust of Norway is a voluntary association, a trust, and a property owner. Since 1845, it has developed expertise in restoration and maintenance of historical monuments. It is based on personal memberships...

  6. 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.

  7. Default values

    International Nuclear Information System (INIS)

    1987-08-01

    In making calculations for the purposes of radiation protection, numerical values for parameters used in the calculations are selected. In some cases, data directly applicable to the set of conditions for which the calculations are to be made are unavailable. Therefore, the selection of the values for these parameters may be based on more general data available from the literature or other sources. These values may be referred to as 'default values', that is, values used in default of those based on directly applicable data. The following policy will be applied by Atomic Energy Control Board (AECB) staff in reviewing the radiation protection aspects of submissions associated with licensing, in participating with other organizations in the development of codes and standards, and in any other work which relies to some extent on using default values

  8. FRET biosensors reveal AKAP-mediated shaping of subcellular PKA activity and a novel mode of Ca(2+)/PKA crosstalk.

    Science.gov (United States)

    Schott, Micah B; Gonowolo, Faith; Maliske, Benjamin; Grove, Bryon

    2016-04-01

    Scaffold proteins play a critical role in cellular homeostasis by anchoring signaling enzymes in close proximity to downstream effectors. In addition to anchoring static enzyme complexes, some scaffold proteins also form dynamic signalosomes that can traffic to different subcellular compartments upon stimulation. Gravin (AKAP12), a multivalent scaffold, anchors PKA and other enzymes to the plasma membrane under basal conditions, but upon [Ca(2+)]i elevation, is rapidly redistributed to the cytosol. Because gravin redistribution also impacts PKA localization, we postulate that gravin acts as a calcium "switch" that modulates PKA-substrate interactions at the plasma membrane, thus facilitating a novel crosstalk mechanism between Ca(2+) and PKA-dependent pathways. To assess this, we measured the impact of gravin-V5/His expression on compartmentalized PKA activity using the FRET biosensor AKAR3 in cultured cells. Upon treatment with forskolin or isoproterenol, cells expressing gravin-V5/His showed elevated levels of plasma membrane PKA activity, but cytosolic PKA activity levels were reduced compared with control cells lacking gravin. This effect required both gravin interaction with PKA and localization at the plasma membrane. Pretreatment with calcium-elevating agents thapsigargin or ATP caused gravin redistribution away from the plasma membrane and prevented gravin from elevating PKA activity levels at the membrane. Importantly, this mode of Ca(2+)/PKA crosstalk was not observed in cells expressing a gravin mutant that resisted calcium-mediated redistribution from the cell periphery. These results reveal that gravin impacts subcellular PKA activity levels through the spatial targeting of PKA, and that calcium elevation modulates downstream β-adrenergic/PKA signaling through gravin redistribution, thus supporting the hypothesis that gravin mediates crosstalk between Ca(2+) and PKA-dependent signaling pathways. Based on these results, AKAP localization dynamics may

  9. FRET biosensors reveal AKAP-mediated shaping of subcellular PKA activity and a novel mode of Ca2+/PKA crosstalk

    Science.gov (United States)

    Schott, Micah; Gonowolo, Faith; Maliske, Ben; Grove, Bryon

    2016-01-01

    Scaffold proteins play a critical role in cellular homeostasis by anchoring signaling enzymes in close proximity to downstream effectors. In addition to anchoring static enzyme complexes, some scaffold proteins also form dynamic signalosomes that can traffic to different subcellular compartments upon stimulation. Gravin (AKAP12), a multivalent scaffold, anchors PKA and other enzymes to the plasma membrane under basal conditions, but upon [Ca2+]i elevation, is rapidly redistributed to the cytosol. Because gravin redistribution also impacts PKA localization, we postulate that gravin acts as a calcium “switch” that modulates PKA-substrate interactions at the plasma membrane, thus facilitating a novel crosstalk mechanism between Ca2+ and PKA-dependent pathways. To assess this, we measured the impact of gravin-V5/His expression on compartmentalized PKA activity using the FRET biosensor AKAR3 in cultured cells. Upon treatment with forskolin or isoproterenol, cells expressing gravin-V5/His showed elevated levels of plasma membrane PKA activity, but cytosolic PKA activity levels were reduced compared with control cells lacking gravin. This effect required both gravin interaction with PKA and localization at the plasma membrane. Pretreatment with calcium-elevating agents thapsigargin or ATP caused gravin redistribution away from the plasma membrane and prevented gravin from elevating PKA activity levels at the membrane. Importantly, this mode of Ca2+/PKA crosstalk was not observed in cells expressing a gravin mutant that resists calcium-mediated redistribution from the cell periphery. These results reveal that gravin impacts subcellular PKA activity levels through the spatial targeting of PKA, and that calcium elevation modulates downstream β-adrenergic/PKA signaling through gravin redistribution, thus supporting the hypothesis that gravin mediates crosstalk between Ca2+ and PKA-dependent signaling pathways. Based on these results, AKAP localization dynamics may

  10. Advances in Spiropyrans/Spirooxazines and Applications Based on Fluorescence Resonance Energy Transfer (FRET with Fluorescent Materials

    Directory of Open Access Journals (Sweden)

    Hongyan Xia

    2017-12-01

    Full Text Available Studies on the following were reviewed: (1 the structure of spiropyrans and spirooxazines (two kinds of spiro compounds under external stimuli and (2 the construction and applications of composite systems based on fluorescence resonance energy transfer (FRET with fluorescent materials. When treated with different stimuli (light, acids and bases, solvents, metal ions, temperature, redox potential, and so on, spiropyrans/spirooxazines undergo transformations between the ring-closed form (SP, the ring-opened merocyanine (MC form, and the protonated ring-opened form (MCH. This is due to the breakage of the spiro C–O bond and the protonation of MC, along with a color change. Various novel, multifunctional materials based on photochromic spiropyrans and spirooxazines have been successfully developed because of the vastly differently physiochemical properties posssed by the SP, MC and MCH forms. Among the three different structural forms, the MC form has been studied most extensively. The MC form not only gives complexes with various inorganic particles, biological molecules, and organic chemicals but also acts as the energy acceptor (of energy from fluorescent molecules during energy transfer processes that take place under proper conditions. Furthermore, spiropyran and spirooxazine compounds exhibit reversible physicochemical property changes under proper stimuli; this provides more advantages compared with other photochromic compounds. Additionally, the molecular structures of spiropyrans and spirooxazines can be easily modified and extended, so better compounds can be obtained to expand the scope of already known applications. Described in detail are: (1 the structural properties of spiropyrans and spirooxazines and related photochromic mechanisms; (2 composite systems based on spiropyrans and spirooxazines, and (3 fluorescent materials which have potential applications in sensing, probing, and a variety of optical elements.

  11. Deep Value

    DEFF Research Database (Denmark)

    Asness, Clifford S.; Liew, John M.; Pedersen, Lasse Heje

    premium. Following these episodes, the value strategy has (1) high average returns; (2) low market betas, but high betas to a global value factor; (3) deteriorating fundamentals; (4) negative news sentiment; (5) selling pressure; (6) increased limits to arbitrage; and (7) increased arbitrage activity...

  12. Forestland values.

    Science.gov (United States)

    John H. Beuter; Ralph J. Alig

    2004-01-01

    This issue of the journal of Forestry is devoted to articles about forestland values. Viewed broadly, natural resources and humans are our two basic resources. An expression of the importance of land as a foundation for forest ecosystems is forestland value. Our attitudes about land and the forest ecosystems that they support have changed considerably in recent years....

  13. 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.

  14. The Value of Value Sets

    DEFF Research Database (Denmark)

    Sløk-Madsen, Stefan Kirkegaard; Christensen, Jesper

    The world over classrooms in business schools are being taught that corporate values can impact performance. The argument is typically that culture matter more than strategy plans and culture can be influenced and indeed changed by a shared corporate value set. While the claim seems intuitively a...... a unique contribution to the effects of investment in shared company values, and to whether agent rationality can be fundamentally changed by committed organizational efforts....

  15. 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.

  16. 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

  17. Value Representations

    DEFF Research Database (Denmark)

    Rasmussen, Majken Kirkegaard; Petersen, Marianne Graves

    2011-01-01

    Stereotypic presumptions about gender affect the design process, both in relation to how users are understood and how products are designed. As a way to decrease the influence of stereotypic presumptions in design process, we propose not to disregard the aspect of gender in the design process......, as the perspective brings valuable insights on different approaches to technology, but instead to view gender through a value lens. Contributing to this perspective, we have developed Value Representations as a design-oriented instrument for staging a reflective dialogue with users. Value Representations...

  18. Unravelling Value

    DEFF Research Database (Denmark)

    Walsh, Matthew

    Paper presented in the symposium “What’s It All Worth? Material Possessions and Value in Past Societies” at 22nd Annual Meeting of the European Association of Archaeologists, Vilnius, Lithuania.......Paper presented in the symposium “What’s It All Worth? Material Possessions and Value in Past Societies” at 22nd Annual Meeting of the European Association of Archaeologists, Vilnius, Lithuania....

  19. 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

  20. Public Value: rethinking value creation

    OpenAIRE

    Meynhardt, Timo; Gomez, Peter; Strathoff, Pepe; Hermann, Carolin

    2014-01-01

    Managers might refute public criticism of their business as an attitude of taking everything for granted in a saturated society, but ignoring Public Value aspects can threaten the success of new products and even the survival of entire firms.

  1. 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.

  2. 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)

  3. 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.

  4. A graphene oxide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of botulinum neurotoxin A (BoNT/A) enzymatic activity.

    Science.gov (United States)

    Shi, Jingyu; Guo, Jiubiao; Bai, Gongxun; Chan, Chunyu; Liu, Xuan; Ye, Weiwei; Hao, Jianhua; Chen, Sheng; Yang, Mo

    2015-03-15

    Botulinum neurotoxins (BoNTs) are among the most potent toxic bacterial proteins for humans, which make them potential agents for bioterrorism. Therefore, an ultrasensitive detection of BoNTs and their active states is in great need as field-deployable systems for anti-terrorism applications. We report the construction of a novel graphene oxide (GO)-peptide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of the BoNT serotype A light chain (BoNT-LcA) protease activity. A green fluorescence protein (GFP) modified SNAP-25 peptide substrate (SNAP-25-GFP) was optimally designed and synthesized with the centralized recognition/cleavage sites. This FRET platform was constructed by covalent immobilization of peptide substrate on GO with BSA passivation which have advantages of low non-specific adsorption and high stability in protein abundant solution. BoNT-LcA can specifically cleave SNAP-25-GFP substrate covalently immobilized on GO to release the fragment with GFP. Based on fluorescence signal recovery measurement, the target BoNT-LcA was detected sensitively and selectively with the linear detection range from 1fg/mL to 1pg/mL. The limit of detection (LOD) for BoNT-LcA is around 1fg/mL. Copyright © 2014 Elsevier B.V. All rights reserved.

  5. Fluisd elastic instability and fretting-wear characteristics of steam generator helical tubes subjected to single-phase external flow and two-phase internal flow

    International Nuclear Information System (INIS)

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

    2004-01-01

    This study investigates the fluid elastic instability characteristics of steam generator (SG) helical type tubes and the safety assessment of the potential for fretting-wear damages caused by foreign object in operating nuclear power plants. The thermal-hydraulic conditions of both tube side and shell side flow fields are predicted by a general purpose computational fluid dynamics code employing the finite volume element modeling. 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 on the effects of coil diameter and the number of turns on the modal and instability characteristics of tubes, which are expressed in terms of the natural frequency, corresponding mode shape and stability ratio. Also, the wear rate of helical type tube caused by foreign object is calculated using the Archard formula and the remaining life of the tube is predicted, and discussed in this study is the effect of the flow velocity and vibration of the tube on the remaining life of the tube. In addition, addressed is the effect of the external pressure on the vibration and fretting wear characteristics of the tube

  6. 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.

  7. 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.

  8. A FRET-based DNA biosensor tracks OmpR-dependent acidification of Salmonella during macrophage infection.

    Directory of Open Access Journals (Sweden)

    Smarajit Chakraborty

    2015-04-01

    Full Text Available In bacteria, one paradigm for signal transduction is the two-component regulatory system, consisting of a sensor kinase (usually a membrane protein and a response regulator (usually a DNA binding protein. The EnvZ/OmpR two-component system responds to osmotic stress and regulates expression of outer membrane proteins. In Salmonella, EnvZ/OmpR also controls expression of another two-component system SsrA/B, which is located on Salmonella Pathogenicity Island (SPI 2. SPI-2 encodes a type III secretion system, which functions as a nanomachine to inject bacterial effector proteins into eukaryotic cells. During the intracellular phase of infection, Salmonella switches from assembling type III secretion system structural components to secreting effectors into the macrophage cytoplasm, enabling Salmonella to replicate in the phagocytic vacuole. Major questions remain regarding how bacteria survive the acidified vacuole and how acidification affects bacterial secretion. We previously reported that EnvZ sensed cytoplasmic signals rather than extracellular ones, as intracellular osmolytes altered the dynamics of a 17-amino-acid region flanking the phosphorylated histidine. We reasoned that the Salmonella cytoplasm might acidify in the macrophage vacuole to activate OmpR-dependent transcription of SPI-2 genes. To address these questions, we employed a DNA-based FRET biosensor ("I-switch" to measure bacterial cytoplasmic pH and immunofluorescence to monitor effector secretion during infection. Surprisingly, we observed a rapid drop in bacterial cytoplasmic pH upon phagocytosis that was not predicted by current models. Cytoplasmic acidification was completely dependent on the OmpR response regulator, but did not require known OmpR-regulated genes such as ompC, ompF, or ssaC (SPI-2. Microarray analysis highlighted the cadC/BA operon, and additional experiments confirmed that it was repressed by OmpR. Acidification was blocked in the ompR null background in a

  9. Add Value

    DEFF Research Database (Denmark)

    Kobbelgaard, Cecilie Elisabeth

    2016-01-01

    "Add Value – kend din kunde" er et brætspil, som giver både offentlige og private virksomheder unikke muligheder for at forbedre deres service overfor kunderne. Spillet giver, på en alternativ og handlingsorienteret måde, mulighed for at blive skarpere på kundeoplevelsen – hvor er der værdi...... at hente, og hvor kan der spares på tid og ressourcer? Dette samtidig med, at kunderne får den oplevelse og service, de forventer. Når I spiller "Add Value – kend din kunde" sættes der fokus på Jeres kundeservice ud fra kundens perspektiv, og det er i alle Jeres kontaktflader med kunden. Lige fra kunden...

  10. Conservation Value

    OpenAIRE

    Tisdell, Clement A.

    2010-01-01

    This paper outlines the significance of the concept of conservation value and discusses ways in which it is determined paying attention to views stemming from utilitarian ethics and from deontological ethics. The importance of user costs in relation to economic decisions about the conservation and use of natural resources is emphasised. Particular attention is given to competing views about the importance of conserving natural resources in order to achieve economic sustainability. This then l...

  11. Technical concept for a greater-confinement-disposal test facility

    International Nuclear Information System (INIS)

    Hunter, P.H.

    1982-01-01

    Greater confinement disposal (GCO) has been defined by the National Low-Level Waste Program as the disposal of low-level waste in such a manner as to provide greater containment of radiation, reduce potential for migration or dispersion or radionuclides, and provide greater protection from inadvertent human and biological intrusions in order to protect the public health and safety. This paper discusses: the need for GCD; definition of GCD; advantages and disadvantages of GCD; relative dose impacts of GCD versus shallow land disposal; types of waste compatible with GCD; objectives of GCD borehole demonstration test; engineering and technical issues; and factors affecting performance of the greater confinement disposal facility

  12. Dietary breadth of grizzly bears in the Greater Yellowstone Ecosystem

    Science.gov (United States)

    Gunther, Kerry A.; Shoemaker, Rebecca; Frey, Kevin L.; Haroldson, Mark A.; Cain, Steven L.; van Manen, Frank T.; Fortin, Jennifer K.

    2014-01-01

    Grizzly bears (Ursus arctos) in the Greater Yellowstone Ecosystem (GYE) are opportunistic omnivores that eat a great diversity of plant and animal species. Changes in climate may affect regional vegetation, hydrology, insects, and fire regimes, likely influencing the abundance, range, and elevational distribution of the plants and animals consumed by GYE grizzly bears. Determining the dietary breadth of grizzly bears is important to document future changes in food resources and how those changes may affect the nutritional ecology of grizzlies. However, no synthesis exists of all foods consumed by grizzly bears in the GYE. We conducted a review of available literature and compiled a list of species consumed by grizzly bears in the GYE. We documented >266 species within 200 genera from 4 kingdoms, including 175 plant, 37 invertebrate, 34 mammal, 7 fungi, 7 bird, 4 fish, 1 amphibian, and 1 algae species as well as 1 soil type consumed by grizzly bears. The average energy values of the ungulates (6.8 kcal/g), trout (Oncorhynchus spp., 6.1 kcal/g), and small mammals (4.5 kcal/g) eaten by grizzlies were higher than those of the plants (3.0 kcal/g) and invertebrates (2.7 kcal/g) they consumed. The most frequently detected diet items were graminoids, ants (Formicidae), whitebark pine seeds (Pinus albicaulis), clover (Trifolium spp.), and dandelion (Taraxacum spp.). The most consistently used foods on a temporal basis were graminoids, ants, whitebark pine seeds, clover, elk (Cervus elaphus), thistle (Cirsium spp.), and horsetail (Equisetum spp.). Historically, garbage was a significant diet item for grizzlies until refuse dumps were closed. Use of forbs increased after garbage was no longer readily available. The list of foods we compiled will help managers of grizzly bears and their habitat document future changes in grizzly bear food habits and how bears respond to changing food resources.

  13. Greater trochanter pain syndrome: A descriptive MR imaging study

    Energy Technology Data Exchange (ETDEWEB)

    Klontzas, Michail E., E-mail: miklontzas@gmail.com; Karantanas, Apostolos H., E-mail: akarantanas@gmail.com

    2014-10-15

    Objective: Greater trochanter pain syndrome (GTPS) is a diverse clinical entity caused by a variety of underlying conditions. We sought to explore the impact of (1) hip morphology, namely the center-edge angle (CEa) and femoral neck-shaft (NSa) angle, (2) hip abductor tendon degeneration, (3) the dimensions of peritrochanteric edema and (4) bursitis, on the presence of GTPS, using MR imaging. Materials and methods: The presence of pain was prospectively assessed blindly by the senior author. CEa and NSa were blindly measured in 174 hip MR examinations, after completion of the clinical evaluation by another evaluator. The existence and dimensions of T2 hyperintensity of the peritrochanteric soft tissues, the existence and dimensions of bursae, as well as degeneration and tearing of gluteus tendons were also recorded. Results: Out of 174 examinations, 91 displayed peritrochanteric edema (group A) and 34 bursitis, all with peritrochanteric edema (group B). A number of 78 patients from both A and B groups, showed gluteus medius tendon degeneration and one tendon tear. CEa of groups A and B were 6° higher than those of normals (group C, P = 0.0038). The mean age of normals was 16.6 years less than in group A and 19.8 years less than in group B (P < 0.0001). Bursitis was associated with pain with a negative predictive value of 97% (P = 0.0003). Conclusion: Acetabular morphology is associated with GTPS and the absence of bursitis was proved to be clinically relevant. Peritrochanteric edema alone was not associated with local pain.

  14. Greater trochanter pain syndrome: A descriptive MR imaging study

    International Nuclear Information System (INIS)

    Klontzas, Michail E.; Karantanas, Apostolos H.

    2014-01-01

    Objective: Greater trochanter pain syndrome (GTPS) is a diverse clinical entity caused by a variety of underlying conditions. We sought to explore the impact of (1) hip morphology, namely the center-edge angle (CEa) and femoral neck-shaft (NSa) angle, (2) hip abductor tendon degeneration, (3) the dimensions of peritrochanteric edema and (4) bursitis, on the presence of GTPS, using MR imaging. Materials and methods: The presence of pain was prospectively assessed blindly by the senior author. CEa and NSa were blindly measured in 174 hip MR examinations, after completion of the clinical evaluation by another evaluator. The existence and dimensions of T2 hyperintensity of the peritrochanteric soft tissues, the existence and dimensions of bursae, as well as degeneration and tearing of gluteus tendons were also recorded. Results: Out of 174 examinations, 91 displayed peritrochanteric edema (group A) and 34 bursitis, all with peritrochanteric edema (group B). A number of 78 patients from both A and B groups, showed gluteus medius tendon degeneration and one tendon tear. CEa of groups A and B were 6° higher than those of normals (group C, P = 0.0038). The mean age of normals was 16.6 years less than in group A and 19.8 years less than in group B (P < 0.0001). Bursitis was associated with pain with a negative predictive value of 97% (P = 0.0003). Conclusion: Acetabular morphology is associated with GTPS and the absence of bursitis was proved to be clinically relevant. Peritrochanteric edema alone was not associated with local pain

  15. Assessing Human Impacts on the Greater Akaki River, Ethiopia ...

    African Journals Online (AJOL)

    We assessed the impacts of human activities on the Greater Akaki River using physicochemical parameters and macroinvertebrate metrics. Physicochemical samples and macroinvertebrates were collected bimonthly from eight sites established on the Greater Akaki River from February 2006 to April 2006. Eleven metrics ...

  16. Comparative Education in Greater China: Contexts, Characteristics, Contrasts and Contributions.

    Science.gov (United States)

    Bray, Mark; Qin, Gui

    2001-01-01

    The evolution of comparative education in Greater China (mainland China, Taiwan, Hong Kong, and Macau) has been influenced by size, culture, political ideologies, standard of living, and colonialism. Similarities and differences in conceptions of comparative education are identified among the four components and between Greater China and other…

  17. Greater temperature sensitivity of plant phenology at colder sites

    DEFF Research Database (Denmark)

    Prevey, Janet; Vellend, Mark; Ruger, Nadja

    2017-01-01

    Warmer temperatures are accelerating the phenology of organisms around the world. Temperature sensitivity of phenology might be greater in colder, higher latitude sites than in warmer regions, in part because small changes in temperature constitute greater relative changes in thermal balance...

  18. Breeding of Greater and Lesser Flamingos at Sua Pan, Botswana ...

    African Journals Online (AJOL)

    to fledging was unknown owing to the rapid drying of the pan in late March 1999. No Greater Flamingo breeding was seen that season. Exceptional flooding during 1999–2000 produced highly favourable breeding conditions, with numbers of Greater and Lesser Flamingos breeding estimated to be 23 869 and 64 287 pairs, ...

  19. Surgical anatomy of greater occipital nerve and its relation to ...

    African Journals Online (AJOL)

    Introduction: The knowledge of the anatomy of greater occipital nerve and its relation to occipital artery is important for the surgeon. Blockage or surgical release of greater occipital nerve is clinically effective in reducing or eliminating chronic migraine symptoms. Aim: The aim of this research was to study the anatomy of ...

  20. Surgical anatomy of greater occipital nerve and its relation to ...

    African Journals Online (AJOL)

    Nancy Mohamed El Sekily

    2014-08-19

    Aug 19, 2014 ... Abstract Introduction: The knowledge of the anatomy of greater occipital nerve and its relation to occipital artery is important for the surgeon. Blockage or surgical release of greater occipital nerve is clinically effective in reducing or eliminating chronic migraine symptoms. Aim: The aim of this research was to ...

  1. INDUSTRIAL LAND DEVELOPMENT AND MANUFACTURING DECONCENTRATION IN GREATER JAKARTA

    NARCIS (Netherlands)

    Hudalah, Delik; Viantari, Dimitra; Firman, Tommy; Woltjer, Johan

    2013-01-01

    Industrial land development has become a key feature of urbanization in Greater Jakarta, one of the largest metropolitan areas in Southeast Asia. Following Suharto's market-oriented policy measures in the late 1980s, private developers have dominated the land development projects in Greater Jakarta.

  2. Strategies for Talent Management: Greater Philadelphia Companies in Action

    Science.gov (United States)

    Council for Adult and Experiential Learning (NJ1), 2008

    2008-01-01

    Human capital is one of the critical issues that impacts the Greater Philadelphia region's ability to grow and prosper. The CEO Council for Growth (CEO Council) is committed to ensuring a steady and talented supply of quality workers for this region. "Strategies for Talent Management: Greater Philadelphia Companies in Action" provides…

  3. Secure equality and greater-than tests with sublinear online complexity

    DEFF Research Database (Denmark)

    Lipmaa, Helger; Toft, Tomas

    2013-01-01

    Secure multiparty computation (MPC) allows multiple parties to evaluate functions without disclosing the private inputs. Secure comparisons (testing equality and greater-than) are important primitives required by many MPC applications. We propose two equality tests for ℓ-bit values with O(1) online...

  4. Valuing hope.

    Science.gov (United States)

    McMillan, John; Walker, Simon; Hope, Tony

    2014-01-01

    This article argues that hope is of value in clinical ethics and that it can be important for clinicians to be sensitive to both the risks of false hope and the importance of retaining hope. However, this sensitivity requires an understanding of the complexity of hope and how it bears on different aspects of a well-functioning doctor-patient relationship. We discuss hopefulness and distinguish it, from three different kinds of hope, or 'hopes for', and then relate these distinctions back to differing accounts of autonomy. This analysis matters because it shows how an overly narrow view of the ethical obligations of a clinician to their patient, and autonomy, might lead to scenarios where patients regret the choices they make.

  5. Radiology's value chain.

    Science.gov (United States)

    Enzmann, Dieter R

    2012-04-01

    A diagnostic radiology value chain is constructed to define its main components, all of which are vulnerable to change, because digitization has caused disaggregation of the chain. Some components afford opportunities to improve productivity, some add value, while some face outsourcing to lower labor cost and to information technology substitutes, raising commoditization risks. Digital image information, because it can be competitive at smaller economies of scale, allows faster, differential rates of technological innovation of components, initiating a centralization-to-decentralization technology trend. Digitization, having triggered disaggregation of radiology's professional service model, may soon usher in an information business model. This means moving from a mind-set of "reading images" to an orientation of creating and organizing information for greater accuracy, faster speed, and lower cost in medical decision making. Information businesses view value chain investments differently than do small professional services. In the former model, producing a better business product will extend image interpretation beyond a radiologist's personal fund of knowledge to encompass expanding external imaging databases. A follow-on expansion with integration of image and molecular information into a report will offer new value in medical decision making. Improved interpretation plus new integration will enrich and diversify radiology's key service products, the report and consultation. A more robust, information-rich report derived from a "systems" and "computational" radiology approach will be facilitated by a transition from a professional service to an information business. Under health care reform, radiology will transition its emphasis from volume to greater value. Radiology's future brightens with the adoption of a philosophy of offering information rather than "reads" for decision making. Staunchly defending the status quo via turf wars is unlikely to constitute a

  6. Fair Value or Market Value?

    Directory of Open Access Journals (Sweden)

    Bogdan Cosmin Gomoi

    2014-12-01

    Full Text Available When taking into consideration the issue of defining the “fair value” concept, those less experimented in the area often fall in the “price trap”, which is considered as an equivalent of the fair value of financial structures. This valuation basis appears as a consequence of the trial to provide an “accurate image” by the financial statements and, also, as an opportunity for the premises offered by the activity continuing principle. The specialized literature generates ample controversies regarding the “fair value” concept and the “market value” concept. The paper aims to debate this issue, taking into account various opinions.

  7. Fractures of the greater trochanter following total hip replacement.

    Science.gov (United States)

    Brun, Ole-Christian L; Maansson, Lukas

    2013-01-01

    We studied the incidence of greater trochanteric fractures at our department following THR. In all we examined 911 patients retrospectively and found the occurance of a greater trochanteric fracture to be 3%. Patients with fractures had significantly poorer outcome on Oxford Hip score, Pain VAS, Satisfaction VAS and EQ-5D compared to THR without fractures. Greater trochanteric fracture following THR is one of the most common complications following THR. It has previously been thought to have little impact on the overall outcome following THR, but our study suggests otherwise.

  8. In vivo FRET imaging revealed a regulatory role of RanGTP in kinetochore-microtubule attachments via Aurora B kinase.

    Directory of Open Access Journals (Sweden)

    Yoke-Peng Lee

    Full Text Available Under the fluctuating circumstances provided by the innate dynamics of microtubules and opposing tensions resulted from microtubule-associated motors, it is vital to ensure stable kinetochore-microtubule attachments for accurate segregation. However, a comprehensive understanding of how this regulation is mechanistically achieved remains elusive. Using our newly designed live cell FRET time-lapse imaging, we found that post-metaphase RanGTP is crucial in the maintenance of stable kinetochore-microtubule attachments by regulating Aurora B kinase via the NES-bearing Mst1. More importantly, our study demonstrates that by ensuring stable alignment of metaphase chromosomes prior to segregation, RanGTP is indispensible in governing the genomic integrity and the fidelity of cell cycle progression. Our findings suggest an additional role of RanGTP beyond its known function in mitotic spindle assembly during the prometaphase-metaphase transition.

  9. Valuing vaccination.

    Science.gov (United States)

    Bärnighausen, Till; Bloom, David E; Cafiero-Fonseca, Elizabeth T; O'Brien, Jennifer Carroll

    2014-08-26

    Vaccination has led to remarkable health gains over the last century. However, large coverage gaps remain, which will require significant financial resources and political will to address. In recent years, a compelling line of inquiry has established the economic benefits of health, at both the individual and aggregate levels. Most existing economic evaluations of particular health interventions fail to account for this new research, leading to potentially sizable undervaluation of those interventions. In line with this new research, we set forth a framework for conceptualizing the full benefits of vaccination, including avoided medical care costs, outcome-related productivity gains, behavior-related productivity gains, community health externalities, community economic externalities, and the value of risk reduction and pure health gains. We also review literature highlighting the magnitude of these sources of benefit for different vaccinations. Finally, we outline the steps that need to be taken to implement a broad-approach economic evaluation and discuss the implications of this work for research, policy, and resource allocation for vaccine development and delivery.

  10. The outcome of endoscopy for recalcitrant greater trochanteric pain syndrome.

    Science.gov (United States)

    Drummond, James; Fary, Camdon; Tran, Phong

    2016-11-01

    Greater trochanteric pain syndrome (GTPS), previously referred as trochanteric bursitis, is a debilitating condition characterised by chronic lateral hip pain. The syndrome is thought to relate to gluteal tendinopathy, with most cases responding to non-operative treatment. A number of open and endoscopic surgical techniques targeting the iliotibial band, trochanteric bursa and gluteal tendons have, however, been described for severe recalcitrant cases. We report the outcomes of one such endoscopic approach here. We retrospectively reviewed 49 patients (57 operations) who had undergone endoscopic longitudinal vertical iliotibial band release and trochanteric bursectomy. Inclusion criteria included diagnosed GTPS with a minimum of six months of non-operative treatment. Exclusion criteria included concomitant intra- or extra-articular hip pathology and previous hip surgery including total hip arthroplasty. Outcomes were assessed using the Visual Analogue Scale, Oxford hip Score and International Hip Outcome Tool (iHOT-33). The series included 42 females and 7 males with a mean age of 65.0 years (26.7-88.6). Mean follow-up time was 20.7 months (5.3-41.2). Eight patients had full thickness gluteal tendon tears, of which 7 were repaired. Adjuvant PRP was injected intraoperatively in 38 of 57 operations (67.2 %). At follow-up, overall mean Visual Analogue Scale values had decreased from 7.8 to 2.8 (p < 0.001), Oxford hip Scores had increased from 20.4 to 37.3 (p < 0.001) and iHOT-33 scores had increased from 23.8 to 70.2 (p < 0.001). Of the 57 operations performed, patients reported feeling very satisfied with the surgical outcome in 28 operations (49.1 %), satisfied in 17 operations (29.8 %) and less than satisfied in 12 operations (21.1 %). While the majority of patients with GTPS will improve with non-operative management, endoscopic iliotibial band release, trochanteric bursectomy and gluteal tendon repair is a safe and effective treatment for severe

  11. Prey selection by a reintroduced lion population in the Greater ...

    African Journals Online (AJOL)

    Prey selection by a reintroduced lion population in the Greater Makalali Conservancy, South Africa. Dave Druce, Heleen Genis, Jonathan Braak, Sophie Greatwood, Audrey Delsink, Ross Kettles, Luke Hunter, Rob Slotow ...

  12. LiveDiverse: Case study area, Greater Kruger South Africa

    CSIR Research Space (South Africa)

    Nortje, Karen

    2011-01-01

    Full Text Available Livelihoods and Biodiversity in Developing Countries Case study area: Greater Kruger, South Africa January 2011 Kolhapur, India Where are we? HARDSHIP LIVELIHOODS NATURE & BIODIVERSITY BELIEFS & CULTURAL PRACTISE threesansinv foursansinv onesansinv...

  13. Exploration of the Energy Efficiency of the Greater London Authority ...

    African Journals Online (AJOL)

    GLA Building/City Hall) ... Journal Home > Vol 11, No 2 (2007) > ... The Greater London Authority building was acclaimed as being energy efficient, with claims of 75 % reduction in its annual energy consumption compared to a high specification ...

  14. Molecular insights into the biology of Greater Sage-Grouse

    Science.gov (United States)

    Oyler-McCance, Sara J.; Quinn, Thomas W.

    2011-01-01

    Recent research on Greater Sage-Grouse (Centrocercus urophasianus) genetics has revealed some important findings. First, multiple paternity in broods is more prevalent than previously thought, and leks do not comprise kin groups. Second, the Greater Sage-Grouse is genetically distinct from the congeneric Gunnison sage-grouse (C. minimus). Third, the Lyon-Mono population in the Mono Basin, spanning the border between Nevada and California, has unique genetic characteristics. Fourth, the previous delineation of western (C. u. phaios) and eastern Greater Sage-Grouse (C. u. urophasianus) is not supported genetically. Fifth, two isolated populations in Washington show indications that genetic diversity has been lost due to population declines and isolation. This chapter examines the use of molecular genetics to understand the biology of Greater Sage-Grouse for the conservation and management of this species and put it into the context of avian ecology based on selected molecular studies.

  15. Greater saphenous vein anomaly and aneurysm with subsequent pulmonary embolism

    OpenAIRE

    Ma, Truong; Kornbau, Craig

    2017-01-01

    Abstract Venous aneurysms often present as painful masses. They can present either in the deep or superficial venous system. Deep venous system aneurysms have a greater risk of thromboembolism. Though rare, there have been case reports of superficial aneurysms and thrombus causing significant morbidity such as pulmonary embolism. We present a case of an anomalous greater saphenous vein connection with an aneurysm and thrombus resulting in a pulmonary embolism. This is the only reported case o...

  16. GREATER OMENTUM: MORPHOFUNCTIONAL CHARACTERISTICS AND CLINICAL SIGNIFICANCE IN PEDIATRICS

    Directory of Open Access Journals (Sweden)

    A.V. Nekrutov

    2007-01-01

    Full Text Available The review analyzes the structure organization and pathophysiological age specificities of the greater omentum, which determine its uniqueness and functional diversity in a child's organism. the article discusses protective functions of the organ, its role in the development of post operative complications of children, and the usage in children's reconstructive plastic surgery.Key words: greater omentum, omentitis, of post operative complications, children.

  17. Socio-economic considerations of cleaning Greater Vancouver's air

    International Nuclear Information System (INIS)

    2005-08-01

    Socio-economic considerations of better air quality on the Greater Vancouver population and economy were discussed. The purpose of the study was to provide socio-economic information to staff and stakeholders of the Greater Vancouver Regional District (GVRD) who are participating in an Air Quality Management Plan (AQMP) development process and the Sustainable Region Initiative (SRI) process. The study incorporated the following methodologies: identification and review of Canadian, American, and European quantitative socio-economic, cost-benefit, cost effectiveness, competitiveness and health analyses of changes in air quality and measures to improve air quality; interviews with industry representatives in Greater Vancouver on competitiveness impacts of air quality changes and ways to improve air quality; and a qualitative analysis and discussion of secondary quantitative information that identifies and evaluates socio-economic impacts arising from changes in Greater Vancouver air quality. The study concluded that for the Greater Vancouver area, the qualitative analysis of an improvement in Greater Vancouver air quality shows positive socio-economic outcomes, as high positive economic efficiency impacts are expected along with good social quality of life impacts. 149 refs., 30 tabs., 6 appendices

  18. Improving greater trochanteric reattachment with a novel cable plate system.

    Science.gov (United States)

    Baril, Yannick; Bourgeois, Yan; Brailovski, Vladimir; Duke, Kajsa; Laflamme, G Yves; Petit, Yvan

    2013-03-01

    Cable-grip systems are commonly used for greater trochanteric reattachment because they have provided the best fixation performance to date, even though they have a rather high complication rate. A novel reattachment system is proposed with the aim of improving fixation stability. It consists of a Y-shaped fixation plate combined with locking screws and superelastic cables to reduce cable loosening and limit greater trochanter movement. The novel system is compared with a commercially available reattachment system in terms of greater trochanter movement and cable tensions under different greater trochanteric abductor application angles. A factorial design of experiments was used including four independent variables: plate system, cable type, abductor application angle, and femur model. The test procedure included 50 cycles of simultaneous application of an abductor force on the greater trochanter and a hip force on the femoral head. The novel plate reduces the movements of a greater trochanter fragment within a single loading cycle up to 26%. Permanent degradation of the fixation (accumulated movement based on 50-cycle testing) is reduced up to 46%. The use of superelastic cables reduces tension loosening up to 24%. However this last improvement did not result in a significant reduction of the grater trochanter movement. The novel plate and cables present advantages over the commercially available greater trochanter reattachment system. The plate reduces movements generated by the hip abductor. The superelastic cables reduce cable loosening during cycling. Both of these positive effects could decrease the risks related to grater trochanter non-union. Copyright © 2012 IPEM. Published by Elsevier Ltd. All rights reserved.

  19. Technical concept for a Greater Confinement Disposal test facility

    International Nuclear Information System (INIS)

    Hunter, P.H.

    1982-01-01

    For the past two years, Ford, Bacon and Davis has been performing technical services for the Department of Energy at the Nevada Test Site in specific development of defense low-level waste management concepts for greater confinement disposal concept with particular application to arid sites. The investigations have included the development of Criteria for Greater Confinement Disposal, NVO-234, which was published in May of 1981 and the draft of the technical concept for Greater Confinement Disposal, with the latest draft published in November 1981. The final draft of the technical concept and design specifications are expected to be published imminently. The document is prerequisite to the actual construction and implementation of the demonstration facility this fiscal year. The GCD Criteria Document, NVO-234 is considered to contain information complimentary and compatible with that being developed for the reserved section 10 CFR 61.51b of the NRCs proposed licensing rule for low level waste disposal facilities

  20. Expatriate job performance in Greater China: Does age matter?

    DEFF Research Database (Denmark)

    Selmer, Jan; Lauring, Jakob; Feng, Yunxia

    to expatriates in Chinese societies. It is possible that older business expatriates will receive more respect and be treated with more deference in a Chinese cultural context than their apparently younger colleagues. This may have a positive impact on expatriates’ job performance. To empirically test...... this presumption, business expatriates in Greater Chine were targeted by a survey. Controlling for the potential bias of a number of background variables, results indicate that contextual/managerial performance, including general managerial functions applied to the subsidiary in Greater China, had a positive...

  1. Absenteeism movement in Greater Poland in 1840–1902

    OpenAIRE

    Izabela Krasińska

    2013-01-01

    The article presents the origins and development of the idea of absenteeism in Greater Poland in the 19th century. The start date for the research is 1840, which is considered to be a breakthrough year in the history of an organized absenteeism movement in Greater Poland. It was due to the Association for the Suppression of the Use of Vodka (Towarzystwo ku Przytłumieniu Używania Wódki) in the Great Duchy of Posen that was then established in Kórnik. It was a secular organization that came int...

  2. Adjustment of Business Expatriates in Greater China: A Strategic Perspective

    DEFF Research Database (Denmark)

    Selmer, Jan

    2006-01-01

    Research has found that due to similarities, firms which have gained business experience elsewhere in Greater China may exhibit relatively better performance in mainland China. Hence, the experience of business expatriates could be of strategic importance for the expansion path of their firms...

  3. College Students with ADHD at Greater Risk for Sleep Disorders

    Science.gov (United States)

    Gaultney, Jane F.

    2014-01-01

    The pediatric literature indicates that children with ADHD are at greater risk for sleep problems, daytime sleepiness, and some sleep disorders than children with no diagnosed disability. It has not been determined whether this pattern holds true among emerging adults, and whether comorbid sleep disorders with ADHD predict GPA. The present study…

  4. Ecology of greater sage-grouse in the Dakotas

    Science.gov (United States)

    Christopher C. Swanson

    2009-01-01

    Greater sage-grouse (Centrocercus urophasianus) populations and the sagebrush (Artemisia spp.) communities that they rely on have dramatically declined from historic levels. Moreover, information regarding sage-grouse annual life-history requirements at the eastern-most extension of sagebrush steppe communities is lacking....

  5. The Easterlin Illusion: Economic growth does go with greater happiness

    NARCIS (Netherlands)

    R. Veenhoven (Ruut); F. Vergunst (Floris)

    2014-01-01

    markdownabstract__Abstract__ The 'Easterlin Paradox' holds that economic growth in nations does not buy greater happiness for the average citizen. This thesis was advanced in the 1970s on the basis of the then available data on happiness in nations. Later data have disproved most of the empirical

  6. Job-Sharing at the Greater Victoria Public Library.

    Science.gov (United States)

    Miller, Don

    1978-01-01

    Describes the problems associated with the management of part-time library employees and some solutions afforded by a job sharing arrangement in use at the Greater Victoria Public Library. This is a voluntary work arrangement, changing formerly full-time positions into multiple part-time positions. (JVP)

  7. Radiographic features of tuberculous osteitis in greater trochanter and lschium

    International Nuclear Information System (INIS)

    Hahm, So Hee; Lee, Ye Ri; Kim, Dong Jin; Sung, Ki Jun; Lim, Jong Nam

    1996-01-01

    To evaluate, if possible, the radiographic features of tuberculous osteitis in the greater trochanter and ischium, and to determine the cause of the lesions. We reterospectively reviewed the plain radiographic findings of 14 ptients with histologically proven tuberculous osteitis involving the greater trochanter and ischium. In each case, the following were analyzed:morphology of bone destruction, including cortical erosion;periosteal reaction;presence or abscence of calcific shadows in adjacent soft tissue. On the basis of an analysis of radiographic features and correlation of the anatomy with adjacent structures we attempted to determine causes. Of the 14 cases evaluated, 12 showed varrious degrees of extrinsic erosion on the outer cortical bone of the greater trochanter and ischium ; in two cases, bone destruction was so severe that the radiographic features of advanced perforated osteomyelitis were simulated. In addition to findings of bone destruction, in these twelve cases, the presence of sequestrum or calcific shadows was seen in adjacent soft tissue. Tuberculous osteitis in the greater trochanter and ischium showed the characteristic findings of chronic extrinsic erosion. On the basis of these findings we can suggest that these lesions result from an extrinsic pathophysiologic cause such as adjacent bursitis

  8. Radiographic features of tuberculous osteitis in greater trochanter and lschium

    Energy Technology Data Exchange (ETDEWEB)

    Hahm, So Hee; Lee, Ye Ri [Hanil Hospital Affiliated to KEPCO, Seoul (Korea, Republic of); Kim, Dong Jin; Sung, Ki Jun [Yonsei Univ. Wonju College of Medicine, Wonju (Korea, Republic of); Lim, Jong Nam [Konkuk Univ. College of Medicine, Seoul (Korea, Republic of)

    1996-11-01

    To evaluate, if possible, the radiographic features of tuberculous osteitis in the greater trochanter and ischium, and to determine the cause of the lesions. We reterospectively reviewed the plain radiographic findings of 14 ptients with histologically proven tuberculous osteitis involving the greater trochanter and ischium. In each case, the following were analyzed:morphology of bone destruction, including cortical erosion;periosteal reaction;presence or abscence of calcific shadows in adjacent soft tissue. On the basis of an analysis of radiographic features and correlation of the anatomy with adjacent structures we attempted to determine causes. Of the 14 cases evaluated, 12 showed varrious degrees of extrinsic erosion on the outer cortical bone of the greater trochanter and ischium ; in two cases, bone destruction was so severe that the radiographic features of advanced perforated osteomyelitis were simulated. In addition to findings of bone destruction, in these twelve cases, the presence of sequestrum or calcific shadows was seen in adjacent soft tissue. Tuberculous osteitis in the greater trochanter and ischium showed the characteristic findings of chronic extrinsic erosion. On the basis of these findings we can suggest that these lesions result from an extrinsic pathophysiologic cause such as adjacent bursitis.

  9. Greater Confinement Disposal trench and borehole operations status

    International Nuclear Information System (INIS)

    Harley, J.P. Jr.; Wilhite, E.L.; Jaegge, W.J.

    1987-01-01

    Greater Confinement Disposal (GCD) facilities have been constructed within the operating burial ground at the Savannah River Plant (SRP) to dispose of the higher activity fraction of SRP low-level waste. GCD practices of waste segregation, packaging, emplacement below the root zone, and waste stabilization are being used in the demonstration. 2 refs., 2 figs., 2 tabs

  10. The Mesozoic-Cenozoic tectonic evolution of the Greater Caucasus

    NARCIS (Netherlands)

    Saintot, A.N.; Brunet, M.F.; Yakovlev, F.; Sébrier, M.; Stephenson, R.A.; Ershov, A.V.; Chalot-Prat, F.; McCann, T.

    2006-01-01

    The Greater Caucasus (GC) fold-and-thrust belt lies on the southern deformed edge of the Scythian Platform (SP) and results from the Cenoozoic structural inversion of a deep marine Mesozoic basin in response to the northward displacement of the Transcaucasus (lying south of the GC subsequent to the

  11. Introduction. China and the Challenges in Greater Middle East

    DEFF Research Database (Denmark)

    Sørensen, Camilla T. N.; Andersen, Lars Erslev; Jiang, Yang

    2016-01-01

    This collection of short papers is an outcome of an international conference entitled China and the Challenges in Greater Middle East, organized by the Danish Institute for International Studies and Copenhagen University on 10 November 2015. The conference sought answers to the following questions...

  12. On the Occurrence of Standardized Regression Coefficients Greater than One.

    Science.gov (United States)

    Deegan, John, Jr.

    1978-01-01

    It is demonstrated here that standardized regression coefficients greater than one can legitimately occur. Furthermore, the relationship between the occurrence of such coefficients and the extent of multicollinearity present among the set of predictor variables in an equation is examined. Comments on the interpretation of these coefficients are…

  13. The Educational Afterlife of Greater Britain, 1903-1914

    Science.gov (United States)

    Gardner, Philip

    2012-01-01

    Following its late nineteenth-century emergence as an important element within federalist thinking across the British Empire, the idea of Greater Britain lost much of its political force in the years following the Boer War. The concept however continued to retain considerable residual currency in other fields of Imperial debate, including those…

  14. Professional performance attributes according to accounting entrepreneurs from greater São Paulo

    OpenAIRE

    Ivam Ricardo Peleias; Rafael Augusto Lourenço; Marcos Reinaldo Severino Peters; Carlos Eduardo Farcin Lavarda

    2015-01-01

    Accounting has a broad activity universe, which requires skilled professionals who are prepared to practice the profession. One possibility is to be an accounting entrepreneur, whose profile requires specific attributes and attitudes. The results reported on in this paper derive from a research developed using the oral history technique, concerning the identification and analysis of attributes valued in the performance of the profession by accounting entrepreneurs from greater São Paulo. A fi...

  15. Risk Aversion and the Value of Information.

    Science.gov (United States)

    Eeckhoudt, Louis; Godfroid, Phillippe

    2000-01-01

    Explains why risk aversion does not always induce a greater information value, but instead may induce a lower information value when increased. Presents a basic model defining the concept of perfect information value and providing a numerical illustration. Includes references. (CMK)

  16. Greater-than-Class C low-level radioactive waste characterization. Appendix E-4: Packaging factors for greater-than-Class C low-level radioactive waste

    International Nuclear Information System (INIS)

    Quinn, G.; Grant, P.; Winberg, M.; Williams, K.

    1994-09-01

    This report estimates packaging factors for several waste types that are potential greater-than-Class C (GTCC) low-level radioactive waste (LLW). The packaging factor is defined as the volume of a GTCC LLW disposal container divided by the as-generated or ''unpackaged'' volume of the waste loaded into the disposal container. Packaging factors reflect any processes that reduce or increase an original unpackaged volume of GTCC LLW, the volume inside a waste container not occupied by the waste, and the volume of the waste container itself. Three values are developed that represent (a) the base case or most likely value for a packaging factor, (b) a high case packaging factor that corresponds to the largest anticipated disposal volume of waste, and (c) a low case packaging factor for the smallest volume expected. GTCC LLW is placed in three categories for evaluation in this report: activated metals, sealed sources, and all other waste

  17. Value-based genomics.

    Science.gov (United States)

    Gong, Jun; Pan, Kathy; Fakih, Marwan; Pal, Sumanta; Salgia, Ravi

    2018-03-20

    Advancements in next-generation sequencing have greatly enhanced the development of biomarker-driven cancer therapies. The affordability and availability of next-generation sequencers have allowed for the commercialization of next-generation sequencing platforms that have found widespread use for clinical-decision making and research purposes. Despite the greater availability of tumor molecular profiling by next-generation sequencing at our doorsteps, the achievement of value-based care, or improving patient outcomes while reducing overall costs or risks, in the era of precision oncology remains a looming challenge. In this review, we highlight available data through a pre-established and conceptualized framework for evaluating value-based medicine to assess the cost (efficiency), clinical benefit (effectiveness), and toxicity (safety) of genomic profiling in cancer care. We also provide perspectives on future directions of next-generation sequencing from targeted panels to whole-exome or whole-genome sequencing and describe potential strategies needed to attain value-based genomics.

  18. 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.

  19. Higher motivation - greater control? The effect of arousal on judgement.

    Science.gov (United States)

    Riemer, Hila; Viswanathan, Madhu

    2013-01-01

    This research examines control over the effect of arousal, a dimension of affect, on judgement. Past research shows that high processing motivation enhances control over the effects of affect on judgement. Isolating and studying arousal as opposed to valence, the other dimension of affect, and its effect on judgement, we identify boundary conditions for past findings. Drawing from the literature on processes by which arousal influences judgement, we demonstrate that the role of motivation is contingent upon the type of judgement task (i.e., memory- versus stimulus-based judgement). In stimulus-based judgement, individuals exert greater control over the effect of arousal on judgement under low compared to high motivation. In contrast, in memory-based judgement individuals exert greater control over the effect of arousal under high compared to low motivation. Theoretical implications and avenues for future research are discussed.

  20. Patient expectations predict greater pain relief with joint arthroplasty.

    Science.gov (United States)

    Gandhi, Rajiv; Davey, John Roderick; Mahomed, Nizar

    2009-08-01

    We examined the relationship between patient expectations of total joint arthroplasty and functional outcomes. We surveyed 1799 patients undergoing primary hip or knee arthroplasty for demographic data and Western Ontario McMaster University Osteoarthritis Index scores at baseline, 3 months, and 1 year of follow-up. Patient expectations were determined with 3 survey questions. The patients with the greatest expectations of surgery were younger, male, and had a lower body mass index. Linear regression modeling showed that a greater expectation of pain relief with surgery independently predicted greater reported pain relief at 1 year of follow-up, adjusted for all relevant covariates (P relief after joint arthroplasty is an important predictor of outcomes at 1 year.

  1. Torsion of the greater omentum: A rare preoperative diagnosis

    International Nuclear Information System (INIS)

    Tandon, Ankit Anil; Lim, Kian Soon

    2010-01-01

    Torsion of the greater omentum is a rare acute abdominal condition that is seldom diagnosed preoperatively. We report the characteristic computed tomography (CT) scan findings and the clinical implications of this unusual diagnosis in a 41-year-old man, who also had longstanding right inguinal hernia. Awareness of omental torsion as a differential diagnosis in the acute abdomen setting is necessary for correct patient management

  2. Ecological specialization and morphological diversification in Greater Antillean boas.

    Science.gov (United States)

    Reynolds, R Graham; Collar, David C; Pasachnik, Stesha A; Niemiller, Matthew L; Puente-Rolón, Alberto R; Revell, Liam J

    2016-08-01

    Colonization of islands can dramatically influence the evolutionary trajectories of organisms, with both deterministic and stochastic processes driving adaptation and diversification. Some island colonists evolve extremely large or small body sizes, presumably in response to unique ecological circumstances present on islands. One example of this phenomenon, the Greater Antillean boas, includes both small (<90 cm) and large (4 m) species occurring on the Greater Antilles and Bahamas, with some islands supporting pairs or trios of body-size divergent species. These boas have been shown to comprise a monophyletic radiation arising from a Miocene dispersal event to the Greater Antilles, though it is not known whether co-occurrence of small and large species is a result of dispersal or in situ evolution. Here, we provide the first comprehensive species phylogeny for this clade combined with morphometric and ecological data to show that small body size evolved repeatedly on separate islands in association with specialization in substrate use. Our results further suggest that microhabitat specialization is linked to increased rates of head shape diversification among specialists. Our findings show that ecological specialization following island colonization promotes morphological diversity through deterministic body size evolution and cranial morphological diversification that is contingent on island- and species-specific factors. © 2016 The Author(s). Evolution © 2016 The Society for the Study of Evolution.

  3. Moderate Baseline Vagal Tone Predicts Greater Prosociality in Children

    Science.gov (United States)

    Miller, Jonas G.; Kahle, Sarah; Hastings, Paul D.

    2016-01-01

    Vagal tone is widely believed to be an important physiological aspect of emotion regulation and associated positive behaviors. However, there is inconsistent evidence for relations between children’s baseline vagal tone and their helpful or prosocial responses to others (Hastings & Miller, 2014). Recent work in adults suggests a quadratic association (inverted U-shape curve) between baseline vagal tone and prosociality (Kogan et al., 2014). The present research examined whether this nonlinear association was evident in children. We found consistent evidence for a quadratic relation between vagal tone and prosociality across 3 samples of children using 6 different measures. Compared to low and high vagal tone, moderate vagal tone in early childhood concurrently predicted greater self-reported prosociality (Study 1), observed empathic concern in response to the distress of others and greater generosity toward less fortunate peers (Study 2), and longitudinally predicted greater self-, mother-, and teacher-reported prosociality 5.5 years later in middle childhood (Study 3). Taken together, our findings suggest that moderate vagal tone at rest represents a physiological preparedness or tendency to engage in different forms of prosociality across different contexts. Early moderate vagal tone may reflect an optimal balance of regulation and arousal that helps prepare children to sympathize, comfort, and share with others. PMID:27819463

  4. Absenteeism movement in Greater Poland in 1840–1902

    Directory of Open Access Journals (Sweden)

    Izabela Krasińska

    2013-12-01

    Full Text Available The article presents the origins and development of the idea of absenteeism in Greater Poland in the 19th century. The start date for the research is 1840, which is considered to be a breakthrough year in the history of an organized absenteeism movement in Greater Poland. It was due to the Association for the Suppression of the Use of Vodka (Towarzystwo ku Przytłumieniu Używania Wódki in the Great Duchy of Posen that was then established in Kórnik. It was a secular organization that came into being on an initiative of doctor De La Roch, who was a German surgeon of a French origin. However, as early as 1844, the idea of absenteeism raised an interest of catholic clergymen of Greater Poland with high ranking clergy such as Rev. Leon Michał Przyłuski, Archbishop of Gniezno and Rev. Jan Kanty Dąbrowski, Archbishop of Posen, and later on Archbishops Rev. Mieczysław Halka Ledóchowski and Rev. Florian Oksza Stablewski. They were fascinated with activities of Rev. Jan Nepomucen Fick, Parish Priest of Piekary Śląskie and several other priests on whose initiative a lot of church brotherhoods of so called holy continence were set up in Upper Silesia as early as the first half-year of 1844. It was due to Bishop Dąbrowski that 100 000 people took vows of absenteeism in 1844–1845, becoming members of brotherhoods of absenteeism. In turn, it was an initiative of Archbishop Przyłuski that Jesuit missionaries – Rev. Karol Bołoz Antoniewicz, Rev. Teofil Baczyński and Rev. Kamil Praszałowicz, arrived in Greater Poland from Galicia in 1852 to promote the idea of absenteeism. Starting from 1848, they were helping Silesian clergymen to spread absenteeism. Clergymen of Greater Poland were also active in secular absenteeism associations. They became involved in the workings of the Association for the Promotion of Absenteeism that was set up by Zygmunt Celichowski in Kórnik in 1887, and especially in the Jutrzenka Absenteeism Association

  5. The post-orgasmic prolactin increase following intercourse is greater than following masturbation and suggests greater satiety.

    Science.gov (United States)

    Brody, Stuart; Krüger, Tillmann H C

    2006-03-01

    Research indicates that prolactin increases following orgasm are involved in a feedback loop that serves to decrease arousal through inhibitory central dopaminergic and probably peripheral processes. The magnitude of post-orgasmic prolactin increase is thus a neurohormonal index of sexual satiety. Using data from three studies of men and women engaging in masturbation or penile-vaginal intercourse to orgasm in the laboratory, we report that for both sexes (adjusted for prolactin changes in a non-sexual control condition), the magnitude of prolactin increase following intercourse is 400% greater than that following masturbation. The results are interpreted as an indication of intercourse being more physiologically satisfying than masturbation, and discussed in light of prior research reporting greater physiological and psychological benefits associated with coitus than with any other sexual activities.

  6. Sexual predators, energy development, and conservation in greater Yellowstone.

    Science.gov (United States)

    Berger, Joel; Beckmann, Jon P

    2010-06-01

    In the United States, as elsewhere, a growing debate pits national energy policy and homeland security against biological conservation. In rural communities the extraction of fossil fuels is often encouraged because of the employment opportunities it offers, although the concomitant itinerant workforce is often associated with increased wildlife poaching. We explored possible positive and negative factors associated with energy extraction in the Greater Yellowstone Ecosystem (GYE), an area known for its national parks, intact biological diversity, and some of the New World's longest terrestrial migrations. Specifically, we asked whether counties with different economies-recreation (ski), agrarian (ranching or farming), and energy extractive (petroleum)-differed in healthcare (gauged by the abundance of hospital beds) and in the frequency of sexual predators. The absolute and relative frequency of registered sex offenders grew approximately two to three times faster in areas reliant on energy extraction. Healthcare among counties did not differ. The strong conflation of community dishevel, as reflected by in-migrant sexual predators, and ecological decay in Greater Yellowstone is consistent with patterns seen in similar systems from Ecuador to northern Canada, where social and environmental disarray exist around energy boomtowns. In our case, that groups (albeit with different aims) mobilized campaigns to help maintain the quality of rural livelihoods by protecting open space is a positive sign that conservation can matter, especially in the face of rampant and poorly executed energy extraction projects. Our findings further suggest that the public and industry need stronger regulatory action to instill greater vigilance when and where social factors and land conversion impact biological systems.

  7. Land cover mapping of Greater Mesoamerica using MODIS data

    Science.gov (United States)

    Giri, Chandra; Jenkins, Clinton N.

    2005-01-01

    A new land cover database of Greater Mesoamerica has been prepared using moderate resolution imaging spectroradiometer (MODIS, 500 m resolution) satellite data. Daily surface reflectance MODIS data and a suite of ancillary data were used in preparing the database by employing a decision tree classification approach. The new land cover data are an improvement over traditional advanced very high resolution radiometer (AVHRR) based land cover data in terms of both spatial and thematic details. The dominant land cover type in Greater Mesoamerica is forest (39%), followed by shrubland (30%) and cropland (22%). Country analysis shows forest as the dominant land cover type in Belize (62%), Cost Rica (52%), Guatemala (53%), Honduras (56%), Nicaragua (53%), and Panama (48%), cropland as the dominant land cover type in El Salvador (60.5%), and shrubland as the dominant land cover type in Mexico (37%). A three-step approach was used to assess the quality of the classified land cover data: (i) qualitative assessment provided good insight in identifying and correcting gross errors; (ii) correlation analysis of MODIS- and Landsat-derived land cover data revealed strong positive association for forest (r2 = 0.88), shrubland (r2 = 0.75), and cropland (r2 = 0.97) but weak positive association for grassland (r2 = 0.26); and (iii) an error matrix generated using unseen training data provided an overall accuracy of 77.3% with a Kappa coefficient of 0.73608. Overall, MODIS 500 m data and the methodology used were found to be quite useful for broad-scale land cover mapping of Greater Mesoamerica.

  8. Practicing more retrieval routes leads to greater memory retention.

    Science.gov (United States)

    Zheng, Jun; Zhang, Wei; Li, Tongtong; Liu, Zhaomin; Luo, Liang

    2016-09-01

    A wealth of research has shown that retrieval practice plays a significant role in improving memory retention. The current study focused on one simple yet rarely examined question: would repeated retrieval using two different retrieval routes or using the same retrieval route twice lead to greater long-term memory retention? Participants elaborately learned 22 Japanese-Chinese translation word pairs using two different mediators. Half an hour after the initial study phase, the participants completed two retrieval sessions using either one mediator (Tm1Tm1) or two different mediators (Tm1Tm2). On the final test, which was performed 1week after the retrieval practice phase, the participants received only the cue with a request to report the mediator (M1 or M2) followed by the target (Experiment 1) or only the mediator (M1 or M2) with a request to report the target (Experiment 2). The results of Experiment 1 indicated that the participants who practiced under the Tm1Tm2 condition exhibited greater target retention than those who practiced under the Tm1Tm1 condition. This difference in performance was due to the significant disadvantage in mediator retrieval and decoding of the unpracticed mediator under the Tm1Tm1 condition. Although mediators were provided to participants on the final test in Experiment 2, decoding of the unpracticed mediators remained less effective than decoding of the practiced mediators. We conclude that practicing multiple retrieval routes leads to greater memory retention than focusing on a single retrieval route. Thus, increasing retrieval variability during repeated retrieval practice indeed significantly improves long-term retention in a delay test. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Taino and African maternal heritage in the Greater Antilles.

    Science.gov (United States)

    Bukhari, Areej; Luis, Javier Rodriguez; Alfonso-Sanchez, Miguel A; Garcia-Bertrand, Ralph; Herrera, Rene J

    2017-12-30

    Notwithstanding the general interest and the geopolitical importance of the island countries in the Greater Antilles, little is known about the specific ancestral Native American and African populations that settled them. In an effort to alleviate this lacuna of information on the genetic constituents of the Greater Antilles, we comprehensively compared the mtDNA compositions of Cuba, Dominican Republic, Haiti, Jamaica and Puerto Rico. To accomplish this, the mtDNA HVRI and HVRII regions, as well as coding diagnostic sites, were assessed in the Haitian general population and compared to data from reference populations. The Taino maternal DNA is prominent in the ex-Spanish colonies (61.3%-22.0%) while it is basically non-existent in the ex-French and ex-English colonies of Haiti (0.0%) and Jamaica (0.5%), respectively. The most abundant Native American mtDNA haplogroups in the Greater Antilles are A2, B2 and C1. The African mtDNA component is almost fixed in Haiti (98.2%) and Jamaica (98.5%), and the frequencies of specific African haplogroups vary considerably among the five island nations. The strong persistence of Taino mtDNA in the ex-Spanish colonies (and especially in Puerto Rico), and its absence in the French and English excolonies is likely the result of different social norms regarding mixed marriages with Taino women during the early years after the first contact with Europeans. In addition, this article reports on the results of an integrative approach based on mtDNA analysis and demographic data that tests the hypothesis of a southward shift in raiding zones along the African west coast during the period encompassing the Transatlantic Slave Trade. Copyright © 2017 Elsevier B.V. All rights reserved.

  10. A FRET-based probe for epidermal growth factor receptor bound non-covalently to a pair of synthetic amphipathic helixes

    International Nuclear Information System (INIS)

    Itoh, Reina E.; Kurokawa, Kazuo; Fujioka, Aki; Sharma, Alok; Mayer, Bruce J.; Matsuda, Michiyuki

    2005-01-01

    Epidermal growth factor (EGF) receptor plays a pivotal role in a variety of cellular functions, such as proliferation, differentiation, and migration. To monitor the EGF receptor (EGFR) activity in living cells, we developed a probe for EGFR activity based on the principle of fluorescence resonance energy transfer (FRET). Previously, we developed a probe designated as Picchu (Phosphorylation indicator of the CrkII chimeric unit), which detects the tyrosine phosphorylation of the CrkII adaptor protein. We used a pair of synthetic amphipathic helixes, WinZipA2 and WinZipB1, to bind Picchu non-covalently to the carboxyl-terminus of the EGFR. Using this modified probe named Picchu-Z, the activity of EGFR was followed in EGF-stimulated Cos7 cells. We found that a high level of tyrosine phosphorylation of Picchu-Z probe remained after endocytosis until the point when the EGFR was translocated to the perinuclear region. These findings are in agreement with the previously reported 'signaling endosome' model. Furthermore, by pulse stimulation with EGF and by acute ablation of EGFR activity with AG1478, it was suggested that the phosphorylation of Picchu-Z probe, and probably the phosphorylation of EGFR also, underwent a rapid equilibrium (τ 1/2 < 2 min) between the phosphorylated and dephosphorylated states in the presence of EGF

  11. Monitoring transient elastic energy storage within the rotary motors of single FoF1-ATP synthase by DCO-ALEX FRET

    Science.gov (United States)

    Ernst, Stefan; Düser, Monika G.; Zarrabi, Nawid; Börsch, Michael

    2012-03-01

    The enzyme FoF1-ATP synthase provides the 'chemical energy currency' adenosine triphosphate (ATP) for living cells. Catalysis is driven by mechanochemical coupling of subunit rotation within the enzyme with conformational changes in the three ATP binding sites. Proton translocation through the membrane-bound Fo part of ATP synthase powers a 10-step rotary motion of the ring of c subunits. This rotation is transmitted to the γ and ɛ subunits of the F1 part. Because γ and ɛ subunits rotate in 120° steps, we aim to unravel this symmetry mismatch by real time monitoring subunit rotation using single-molecule Förster resonance energy transfer (FRET). One fluorophore is attached specifically to the F1 motor, another one to the Fo motor of the liposome-reconstituted enzyme. Photophysical artifacts due to spectral fluctuations of the single fluorophores are minimized by a previously developed duty cycle-optimized alternating laser excitation scheme (DCO-ALEX). We report the detection of reversible elastic deformations between the rotor parts of Fo and F1 and estimate the maximum angular displacement during the load-free rotation using Monte Carlo simulations.

  12. A rapid, ratiometric, enzyme-free, and sensitive single-step miRNA detection using three-way junction based FRET probes

    Science.gov (United States)

    Luo, Qingying; Liu, Lin; Yang, Cai; Yuan, Jing; Feng, Hongtao; Chen, Yan; Zhao, Peng; Yu, Zhiqiang; Jin, Zongwen

    2018-03-01

    MicroRNAs (miRNAs) are single stranded endogenous molecules composed of only 18-24 nucleotides which are critical for gene expression regulating the translation of messenger RNAs. Conventional methods based on enzyme-assisted nucleic acid amplification techniques have many problems, such as easy contamination, high cost, susceptibility to false amplification, and tendency to have sequence mismatches. Here we report a rapid, ratiometric, enzyme-free, sensitive, and highly selective single-step miRNA detection using three-way junction assembled (or self-assembled) FRET probes. The developed strategy can be operated within the linear range from subnanomolar to hundred nanomolar concentrations of miRNAs. In comparison with the traditional approaches, our method showed high sensitivity for the miRNA detection and extreme selectivity for the efficient discrimination of single-base mismatches. The results reveal that the strategy paved a new avenue for the design of novel highly specific probes applicable in diagnostics and potentially in microscopic imaging of miRNAs in real biological environments.

  13. FRET structure with non-radiative acceptor provided by dye-linker-glass surface complex and single-molecule photodynamics by TIRFM-polarized imaging

    International Nuclear Information System (INIS)

    Tani, Toshiro; Mashimo, Kei; Suzuki, Tetsu; Horiuchi, Hiromi; Oda, Masaru

    2008-01-01

    We present our recent study of microscopic single-molecule imaging on the artificial complex of tetramethylrhodamine linked with a propyl chain onto silica glass surface, i.e. an asymmetric fluorescence resonance energy transfer (FRET) structure with non-radiative acceptor. In the synthesis of the complex, we used a mixture of two kinds of isomers to introduce rather small photodynamic difference among them. This isomeric structure change will provide more or less a distinctive photophysical change in e.g. non-radiative relaxation rate. Our recent observation at room temperatures, so far, shows that such contributions can be discriminated in the histograms of the fluorescent spot intensities; broad but distinctive multi-components appear. To identify the isomeric difference as a cause of structures, some configurational assumptions are necessary. One such basic prerequisite is that the transition dipoles of the chromophores should be oriented almost parallel to the glass surface. In order to make clear the modeling, we also provide preliminary experiments on the polarization dependence of the imaging under rotating polarization in epi-illumination

  14. Demystifying the Clinical Diagnosis of Greater Trochanteric Pain Syndrome in Women.

    Science.gov (United States)

    Ganderton, Charlotte; Semciw, Adam; Cook, Jill; Pizzari, Tania

    2017-06-01

    To evaluate the diagnostic accuracy of 10 clinical tests that can be used in the diagnosis of greater trochanteric pain syndrome (GTPS) in women, and to compare these clinical tests to magnetic resonance imaging (MRI) findings. Twenty-eight participants with GTPS (49.5 ± 22.0 years) and 18 asymptomatic participants (mean age ± standard deviation [SD], 52.5 ± 22.8 years) were included. A blinded physiotherapist performed 10 pain provocation tests potentially diagnostic for GTPS-palpation of the greater trochanter, resisted external derotation test, modified resisted external derotation test, standard and modified Ober's tests, Patrick's or FABER test, resisted hip abduction, single-leg stance test, and the resisted hip internal rotation test. A sample of 16 symptomatic and 17 asymptomatic women undertook a hip MRI scan. Gluteal tendons were evaluated and categorized as no pathology, mild tendinosis, moderate tendinosis/partial tear, or full-thickness tear. Clinical test analyses show high specificity, high positive predictive value, low to moderate sensitivity, and negative predictive value for most clinical tests. All symptomatic and 88% of asymptomatic participants had pathological gluteal tendon changes on MRI, from mild tendinosis to full-thickness tear. The study found the Patrick's or FABER test, palpation of the greater trochanter, resisted hip abduction, and the resisted external derotation test to have the highest diagnostic test accuracy for GTPS. Tendon pathology on MRI is seen in both symptomatic and asymptomatic women.

  15. Single versus double row suture anchor fixation for greater tuberosity fractures - a biomechanical study.

    Science.gov (United States)

    Seppel, Gernot; Saier, Tim; Martetschläger, Frank; Plath, Johannes E; Guevara-Alvarez, Alberto; Henschel, Julia; Winkler, Martin; Augat, Peter; Imhoff, Andreas B; Buchmann, Stefan

    2017-12-01

    Fractures of the humeral greater tuberosity (GT) are a frequent injury progressively treated with arthroscopic suture anchor repair. Yet, no biomechanical study has been performed comparing fixation strength of arthroscopic single- (SR) vs. double row (DR) fixation. Standardized fractures of the greater tuberosity were created in 12 fresh frozen proximal humeri. After random assignation to the SR or DR group the fixed humeri were tested applying cyclic loading to the supraspinatus and infraspinatus tendon. Load to failure and fragment displacement were assessed by means of an electrodynamic material testing machine using an optical tracking system. Load to failure values were higher in the DR group (649 N; ±176) than in the SR group (490 N; ±145) however without statistical significance (p = .12). In greater tuberosity displacement of 3-5 mm surgical treatment is recommended. The fixing constructs in this study did not reach displacement landmarks of 3 or 5 mm before construct failure as shown in previous studies. Thus the applied traction force (N) at 1 mm displacement was analyzed. In the SR group the load at 1 mm displacement was 277 N; ±46 compared to 260 N; ±62 in the DR group (p = .65). The results suggest that both techniques are viable options for refixation of greater tuberosity fractures. Laboratory study.

  16. Conservation of greater sage-grouse- a synthesis of current trends and future management

    Science.gov (United States)

    Connelly, John W.; Knick, Steven T.; Braun, Clait E.; Baker, William L.; Beever, Erik A.; Christiansen, Thomas J.; Doherty, Kevin E.; Garton, Edward O.; Hagen, Christian A.; Hanser, Steven E.; Johnson, Douglas H.; Leu, Matthias; Miller, Richard F.; Naugle, David E.; Oyler-McCance, Sara J.; Pyke, David A.; Reese, Kerry P.; Schroeder, Michael A.; Stiver, San J.; Walker, Brett L.; Wisdorn, Michael J.

    2011-01-01

    Recent analyses of Greater Sage-Grouse (Centrocercus urophasianus) populations indicate substantial declines in many areas but relatively stable populations in other portions of the species? range. Sagebrush (Artemisia spp.) habitats neces-sary to support sage-grouse are being burned by large wildfires, invaded by nonnative plants, and developed for energy resources (gas, oil, and wind). Management on public lands, which con-tain 70% of sagebrush habitats, has changed over the last 30 years from large sagebrush control projects directed at enhancing livestock grazing to a greater emphasis on projects that often attempt to improve or restore ecological integrity. Never-theless, the mandate to manage public lands to provide traditional consumptive uses as well as recreation and wilderness values is not likely to change in the near future. Consequently, demand and use of resources contained in sagebrush land-scapes plus the associated infrastructure to sup-port increasing human populations in the western United States will continue to challenge efforts to conserve Greater Sage-Grouse. The continued widespread distribution of sage-grouse, albeit at very low densities in some areas, coupled with large areas of important sagebrush habitat that are relatively unaffected by the human footprint, sug-gest that Greater Sage-Grouse populations may be able to persist into the future. We summarize the status of sage-grouse populations and habitats, provide a synthesis of major threats and chal-lenges to conservation of sage-grouse, and suggest a roadmap to attaining conservation goals.

  17. Absorption spectrum of DNA for wavelengths greater than 300 nm

    International Nuclear Information System (INIS)

    Sutherland, J.C.; Griffin, K.P.

    1981-01-01

    Although DNA absorption at wavelengths greater than 300 nm is much weaker than that at shorter wavelengths, this absorption seems to be responsible for much of the biological damage caused by solar radiation of wavelengths less than 320 nm. Accurate measurement of the absorption spectrum of DNA above 300 nm is complicated by turbidity characteristic of concentrated solutions of DNA. We have measured the absorption spectra of DNA from calf thymus, Clostridium perfringens, Escherichia coli, Micrococcus luteus, salmon testis, and human placenta using procedures which separate optical density due to true absorption from that due to turbidity. Above 300 nm, the relative absorption of DNA increases as a function of guanine-cytosine content, presumably because the absorption of guanine is much greater than the absorption of adenine at these wavelengths. This result suggests that the photophysical processes which follow absorption of a long-wavelength photon may, on the average, differ from those induced by shorter-wavelength photons. It may also explain the lower quantum yield for the killing of cells by wavelengths above 300 nm compared to that by shorter wavelengths

  18. Black breast cancer survivors experience greater upper extremity disability.

    Science.gov (United States)

    Dean, Lorraine T; DeMichele, Angela; LeBlanc, Mously; Stephens-Shields, Alisa; Li, Susan Q; Colameco, Chris; Coursey, Morgan; Mao, Jun J

    2015-11-01

    Over one-third of breast cancer survivors experience upper extremity disability. Black women present with factors associated with greater upper extremity disability, including: increased body mass index (BMI), more advanced disease stage at diagnosis, and varying treatment type compared with Whites. No prior research has evaluated the relationship between race and upper extremity disability using validated tools and controlling for these factors. Data were drawn from a survey study among 610 women with stage I-III hormone receptor positive breast cancer. The disabilities of the arm, shoulder and hand (QuickDASH) is an 11-item self-administered questionnaire that has been validated for breast cancer survivors to assess global upper extremity function over the past 7 days. Linear regression and mediation analysis estimated the relationships between race, BMI and QuickDASH score, adjusting for demographics and treatment types. Black women (n = 98) had 7.3 points higher average QuickDASH scores than White (n = 512) women (p disability by 40 %. Even several years post-treatment, Black breast cancer survivors had greater upper extremity disability, which was partially mediated by higher BMIs. Close monitoring of high BMI Black women may be an important step in reducing disparities in cancer survivorship. More research is needed on the relationship between race, BMI, and upper extremity disability.

  19. Greater-confinement disposal of low-level radioactive wastes

    International Nuclear Information System (INIS)

    Trevorrow, L.E.; Gilbert, T.L.; Luner, C.; Merry-Libby, P.A.; Meshkov, N.K.; Yu, C.

    1985-01-01

    Low-level radioactive wastes include a broad spectrum of wastes that have different radionuclide concentrations, half-lives, and physical and chemical properties. Standard shallow-land burial practice can provide adequate protection of public health and safety for most low-level wastes, but a small volume fraction (about 1%) containing most of the activity inventory (approx.90%) requires specific measures known as ''greater-confinement disposal'' (GCD). Different site characteristics and different waste characteristics - such as high radionuclide concentrations, long radionuclide half-lives, high radionuclide mobility, and physical or chemical characteristics that present exceptional hazards - lead to different GCD facility design requirements. Facility design alternatives considered for GCD include the augered shaft, deep trench, engineered structure, hydrofracture, improved waste form, and high-integrity container. Selection of an appropriate design must also consider the interplay between basic risk limits for protection of public health and safety, performance characteristics and objectives, costs, waste-acceptance criteria, waste characteristics, and site characteristics. This paper presents an overview of the factors that must be considered in planning the application of methods proposed for providing greater confinement of low-level wastes. 27 refs

  20. Ratiometric fluorescent sensing of pH values in living cells by dual-fluorophore-labeled i-motif nanoprobes.

    Science.gov (United States)

    Huang, Jin; Ying, Le; Yang, Xiaohai; Yang, Yanjing; Quan, Ke; Wang, He; Xie, Nuli; Ou, Min; Zhou, Qifeng; Wang, Kemin

    2015-09-01

    We designed a new ratiometric fluorescent nanoprobe for sensing pH values in living cells. Briefly, the nanoprobe consists of a gold nanoparticle (AuNP), short single-stranded oligonucleotides, and dual-fluorophore-labeled i-motif sequences. The short oligonucleotides are designed to bind with the i-motif sequences and immobilized on the AuNP surface via Au-S bond. At neutral pH, the dual fluorophores are separated, resulting in very low fluorescence resonance energy transfer (FRET) efficiency. At acidic pH, the i-motif strands fold into a quadruplex structure and leave the AuNP, bringing the dual fluorophores into close proximity, resulting in high FRET efficiency, which could be used as a signal for pH sensing. The nanoprobe possesses abilities of cellular transfection, enzymatic protection, fast response and quantitative pH detection. The in vitro and intracellular applications of the nanoprobe were demonstrated, which showed excellent response in the physiological pH range. Furthermore, our experimental results suggested that the nanoprobe showed excellent spatial and temporal resolution in living cells. We think that the ratiometric sensing strategy could potentially be applied to create a variety of new multicolor sensors for intracellular detection.

  1. The Effects of Work Values, Work-Value Congruence and Work Centrality on Organizational Citizenship Behavior

    OpenAIRE

    Başak Uçanok

    2008-01-01

    The aim of this study is to test the "work values" inventory developed by Tevruz and Turgut and to utilize the concept in a model, which aims to create a greater understanding of the work experience. In the study multiple effects of work values, work-value congruence and work centrality on organizational citizenship behavior are examined. In this respect, it is hypothesized that work values and work-value congruence predict organizational citizenship behavior through work...

  2. [Clinical Results of Endoscopic Treatment of Greater Trochanteric Pain Syndrome].

    Science.gov (United States)

    Zeman, P; Rafi, M; Skala, P; Zeman, J; Matějka, J; Pavelka, T

    2017-01-01

    PURPOSE OF THE STUDY This retrospective study aims to present short-term clinical outcomes of endoscopic treatment of patients with greater trochanteric pain syndrome (GTPS). MATERIAL AND METHODS The evaluated study population was composed of a total of 19 patients (16 women, 3 men) with the mean age of 47 years (19-63 years). In twelve cases the right hip joint was affected, in the remaining seven cases it was the left side. The retrospective evaluation was carried out only in patients with greater trochanteric pain syndrome caused by independent chronic trochanteric bursitis without the presence of m. gluteus medius tear not responding to at least 3 months of conservative treatment. In patients from the followed-up study population, endoscopic trochanteric bursectomy was performed alone or in combination with iliotibial band release. The clinical results were evaluated preoperatively and with a minimum follow-up period of 1 year after the surgery (mean 16 months). The Visual Analogue Scale (VAS) for assessment of pain and WOMAC (Western Ontario MacMaster) score were used. In both the evaluated criteria (VAS and WOMAC score) preoperative and postoperative results were compared. Moreover, duration of surgery and presence of postoperative complications were assessed. Statistical evaluation of clinical results was carried out by an independent statistician. In order to compare the parameter of WOMAC score and VAS pre- and post-operatively the Mann-Whitney Exact Test was used. The statistical significance was set at 0.05. RESULTS The preoperative VAS score ranged 5-9 (mean 7.6) and the postoperative VAS ranged 0-5 (mean 2.3). The WOMAC score ranged 56.3-69.7 (mean 64.2) preoperatively and 79.8-98.3 (mean 89.7) postoperatively. When both the evaluated parameters of VAS and WOMAC score were compared in time, a statistically significant improvement (ppain syndrome yields statistically significant improvement of clinical results with the concurrent minimum incidence of

  3. Use value, exchange value, and resource scarcity

    International Nuclear Information System (INIS)

    Stern, D.I.

    1999-01-01

    The literature on natural resource scarcity indicators is reviewed. Scarcity indicators can be classified by what is being measured: value of the resource stock or value of extracted resource commodities; whose value is considered: social vs. private scarcity; and by the mode of valuation considered: exchange value and use value. Prices and rents are common measures of exchange value or indicators of ''exchange scarcity'' and unit costs can be seen as use value indicators or indicators of u se scarcity . The major aim of this paper is to demonstrate the links between productivity indicators such as unit costs and the classical concept of use value. The two classes of indicator relate to John Commons' discussions of scarcity and efficiency, and a marginal vs. a non-marginal approach to value and scarcity. The classical use value concept also has wider relevance for issues of valuation in energy, resource, and environmental policy. (author)

  4. Strontium isotopic geochemistry of intrusive rocks, Puerto Rico, Greater Antilles

    International Nuclear Information System (INIS)

    Jones, L.M.; Kesler, S.E.

    1980-01-01

    The strontium isotope geochemistry is given for three Puerto Rican intrusive rocks: the granodioritic Morovis and San Lorenzo plutons and the Rio Blanco stock of quartz dioritic composition. The average calculated initial 87 Sr/ 86 Sr ratios are 0.70370, 0.70355 and 0.70408, respectively. In addition, the San Lorenzo data establish a whole-rock isochron of 71 +- 2 m.y., which agrees with the previously reported K-Ar age of 73 m.y. Similarity of most of the intrusive rocks in the Greater Antilles with respect to their strontium isotopic geochemistry regardless of their major element composition indicates that intrusive magmas with a wide range of composition can be derived from a single source material. The most likely source material, in view of the available isotopic data, is the mantle wedge overlying the subduction zone. (orig.)

  5. Slimness is associated with greater intercourse and lesser masturbation frequency.

    Science.gov (United States)

    Brody, Stuart

    2004-01-01

    I examined the relationship of recalled and diary recorded frequency of penile-vaginal intercourse (FSI), noncoital partnered sexual activity, and masturbation to measured waist and hip circumference in 120 healthy adults aged 19-38. Slimmer waist (in men and in the sexes combined) and slimmer hips (in men and women) were associated with greater FSI. Slimmer waist and hips were associated with rated importance of intercourse for men. Noncoital partnered sexual activity had a less consistent association with slimness. Slimmer waist and hips were associated with less masturbation (in men and in the sexes combined). I discuss the results in terms of differences between different sexual behaviors, attractiveness, emotional relatedness, physical sensitivity, sexual dysfunction, sociobiology, psychopharmacological aspects of excess fat and carbohydrate consumption, and implications for sex therapy.

  6. Greater confinement disposal program at the Savannah River Plant

    International Nuclear Information System (INIS)

    Cook, J.R.; Towler, O.A.; Peterson, D.L.; Johnson, G.M.; Helton, B.D.

    1984-01-01

    The first facility to demonstrate Greater Confinement Disposal (GCD) in a humid environment in the United States has been built and is operating at the Savannah River Plant. GCD practices of waste segregation, packaging, emplacement below the root zone, and waste stabilization are being used in the demonstration. Activity concentrations to select wastes for GCD are based on a study of SRP burial records, and are equal to or less than those for Class B waste in 10CFR61. The first disposal units to be constructed are 9-foot diameter, thirty-foot deep boreholes which will be used to dispose of wastes from production reactors, tritiated wastes, and selected wastes from off-site. In 1984 an engineered GCD trench will be constructed for disposal of boxed wastes and large bulky items. 2 figures, 1 table

  7. Evil genius? How dishonesty can lead to greater creativity.

    Science.gov (United States)

    Gino, Francesca; Wiltermuth, Scott S

    2014-04-01

    We propose that dishonest and creative behavior have something in common: They both involve breaking rules. Because of this shared feature, creativity may lead to dishonesty (as shown in prior work), and dishonesty may lead to creativity (the hypothesis we tested in this research). In five experiments, participants had the opportunity to behave dishonestly by overreporting their performance on various tasks. They then completed one or more tasks designed to measure creativity. Those who cheated were subsequently more creative than noncheaters, even when we accounted for individual differences in their creative ability (Experiment 1). Using random assignment, we confirmed that acting dishonestly leads to greater creativity in subsequent tasks (Experiments 2 and 3). The link between dishonesty and creativity is explained by a heightened feeling of being unconstrained by rules, as indicated by both mediation (Experiment 4) and moderation (Experiment 5).

  8. The hydrogen village in the Greater Toronto Area (GTA)

    International Nuclear Information System (INIS)

    Kimmel, T.B.; Smith, R.

    2004-01-01

    'Full text:' A Hydrogen Village (H2V) is a public/private partnership with an objective to accelerate the commercialization of hydrogen and fuel cell technology in Canada and firmly position Canada as the international leader in this sector. The first Hydrogen Village is planned for the Greater Toronto Area (GTA) and will make use of existing hydrogen and fuel cell deployments to assist in its creation. This five year GTA Hydrogen Village program is planned to begin operations in 2004. The Hydrogen Village will demonstrate and deploy various hydrogen production and delivery techniques as well as fuel cells for stationary, transportation (mobile) and portable applications. This paper will provide an overview of the Hydrogen Village and identify the missions, objectives, members and progress within the H2V. (author)

  9. Aerobic Interval Exercise Training Induces Greater Reduction in Cardiac Workload in the Recovery Period in Rats

    International Nuclear Information System (INIS)

    Borges, Juliana Pereira; Masson, Gustavo Santos; Tibiriçá, Eduardo; Lessa, Marcos Adriano

    2014-01-01

    Aerobic interval exercise training has greater benefits on cardiovascular function as compared with aerobic continuous exercise training. The present study aimed at analyzing the effects of both exercise modalities on acute and subacute hemodynamic responses of healthy rats. Thirty male rats were randomly assigned into three groups as follows: continuous exercise (CE, n = 10); interval exercise (IE, n = 10); and control (C, n = 10). Both IE and CE groups performed a 30-minute exercise session. The IE group session consisted of three successive 4-minute periods at 60% of maximal velocity (Max Vel), with 4-minute recovery intervals at 40% of Max Vel. The CE group ran continuously at 50% of Max Vel. Heart rate (HR), blood pressure(BP), and rate pressure product (RPP) were measured before, during and after the exercise session. The CE and IE groups showed an increase in systolic BP and RPP during exercise as compared with the baseline values. After the end of exercise, the CE group showed a lower response of systolic BP and RPP as compared with the baseline values, while the IE group showed lower systolic BP and mean BP values. However, only the IE group had a lower response of HR and RPP during recovery. In healthy rats, one interval exercise session, as compared with continuous exercise, induced similar hemodynamic responses during exercise. However, during recovery, the interval exercise caused greater reductions in cardiac workload than the continuous exercise

  10. Aerobic Interval Exercise Training Induces Greater Reduction in Cardiac Workload in the Recovery Period in Rats

    Science.gov (United States)

    Borges, Juliana Pereira; Masson, Gustavo Santos; Tibiriçá, Eduardo; Lessa, Marcos Adriano

    2014-01-01

    Background Aerobic interval exercise training has greater benefits on cardiovascular function as compared with aerobic continuous exercise training. Objective The present study aimed at analyzing the effects of both exercise modalities on acute and subacute hemodynamic responses of healthy rats. Methods Thirty male rats were randomly assigned into three groups as follows: continuous exercise (CE, n = 10); interval exercise (IE, n = 10); and control (C, n = 10). Both IE and CE groups performed a 30-minute exercise session. The IE group session consisted of three successive 4-minute periods at 60% of maximal velocity (Max Vel), with 4-minute recovery intervals at 40% of Max Vel. The CE group ran continuously at 50% of Max Vel. Heart rate (HR), blood pressure(BP), and rate pressure product (RPP) were measured before, during and after the exercise session. Results The CE and IE groups showed an increase in systolic BP and RPP during exercise as compared with the baseline values. After the end of exercise, the CE group showed a lower response of systolic BP and RPP as compared with the baseline values, while the IE group showed lower systolic BP and mean BP values. However, only the IE group had a lower response of HR and RPP during recovery. Conclusion In healthy rats, one interval exercise session, as compared with continuous exercise, induced similar hemodynamic responses during exercise. However, during recovery, the interval exercise caused greater reductions in cardiac workload than the continuous exercise. PMID:24270864

  11. Aerobic Interval Exercise Training Induces Greater Reduction in Cardiac Workload in the Recovery Period in Rats

    Energy Technology Data Exchange (ETDEWEB)

    Borges, Juliana Pereira, E-mail: julipborges@gmail.com; Masson, Gustavo Santos; Tibiriçá, Eduardo; Lessa, Marcos Adriano [Instituto Oswaldo Cruz - FIOCRUZ, Rio de Janeiro, RJ (Brazil)

    2014-01-15

    Aerobic interval exercise training has greater benefits on cardiovascular function as compared with aerobic continuous exercise training. The present study aimed at analyzing the effects of both exercise modalities on acute and subacute hemodynamic responses of healthy rats. Thirty male rats were randomly assigned into three groups as follows: continuous exercise (CE, n = 10); interval exercise (IE, n = 10); and control (C, n = 10). Both IE and CE groups performed a 30-minute exercise session. The IE group session consisted of three successive 4-minute periods at 60% of maximal velocity (Max Vel), with 4-minute recovery intervals at 40% of Max Vel. The CE group ran continuously at 50% of Max Vel. Heart rate (HR), blood pressure(BP), and rate pressure product (RPP) were measured before, during and after the exercise session. The CE and IE groups showed an increase in systolic BP and RPP during exercise as compared with the baseline values. After the end of exercise, the CE group showed a lower response of systolic BP and RPP as compared with the baseline values, while the IE group showed lower systolic BP and mean BP values. However, only the IE group had a lower response of HR and RPP during recovery. In healthy rats, one interval exercise session, as compared with continuous exercise, induced similar hemodynamic responses during exercise. However, during recovery, the interval exercise caused greater reductions in cardiac workload than the continuous exercise.

  12. The Greater Caucasus Glacier Inventory (Russia, Georgia and Azerbaijan)

    Science.gov (United States)

    Tielidze, Levan G.; Wheate, Roger D.

    2018-01-01

    There have been numerous studies of glaciers in the Greater Caucasus, but none that have generated a modern glacier database across the whole mountain range. Here, we present an updated and expanded glacier inventory at three time periods (1960, 1986, 2014) covering the entire Greater Caucasus. Large-scale topographic maps and satellite imagery (Corona, Landsat 5, Landsat 8 and ASTER) were used to conduct a remote-sensing survey of glacier change, and the 30 m resolution Advanced Spaceborne Thermal Emission and Reflection Radiometer Global Digital Elevation Model (ASTER GDEM; 17 November 2011) was used to determine the aspect, slope and height distribution of glaciers. Glacier margins were mapped manually and reveal that in 1960 the mountains contained 2349 glaciers with a total glacier surface area of 1674.9 ± 70.4 km2. By 1986, glacier surface area had decreased to 1482.1 ± 64.4 km2 (2209 glaciers), and by 2014 to 1193.2 ± 54.0 km2 (2020 glaciers). This represents a 28.8 ± 4.4 % (481 ± 21.2 km2) or 0.53 % yr-1 reduction in total glacier surface area between 1960 and 2014 and an increase in the rate of area loss since 1986 (0.69 % yr-1) compared to 1960-1986 (0.44 % yr-1). Glacier mean size decreased from 0.70 km2 in 1960 to 0.66 km2 in 1986 and to 0.57 km2 in 2014. This new glacier inventory has been submitted to the Global Land Ice Measurements from Space (GLIMS) database and can be used as a basis data set for future studies.

  13. Myiasis in Dogs in the Greater Accra Region of Ghana.

    Science.gov (United States)

    Johnson, Sherry A M; Gakuya, Daniel W; Mbuthia, Paul G; Mande, John D; Afakye, Kofi; Maingi, Ndichu

    2016-01-01

    Myiasis is the infestation of tissues of live vertebrate animals and humans with dipterous larvae. In sub-Saharan Africa, Cordylobia anthropohaga and Cordylobia rodhaini are known to be responsible for cutaneous myiasis in animals and humans. Human cases of myiasis, purportedly acquired in Ghana but diagnosed in other countries, have been reported; however, published data on its occurrence in animals in Ghana is unavailable. This study assessed the prevalence of canine myiasis among owned dogs in the Greater Accra region (GAR) of Ghana. A cross-sectional study was conducted in the Greater Accra region of Ghana, selected for being the region with the highest estimated population density of owned dogs. Physical examination and demographic characteristics of the study dogs were assessed. Management of the dogs was assessed through a questionnaire administered to the dog owners. A total of 392 owned dogs were sampled. Twenty-nine (7.4%) had cutaneous myiasis caused by C. rodhaini. In addition, one (0.2%) of the dogs had intestinal myiasis, with Dermatobia hominis as the offending larvae. Among the breeds of dogs with myiasis, the mongrel was most affected, with 24 (82.8%) out of the 29 cases. The mongrels, majority of which (24; 82.8%) were males, were left to roam freely in the community. Results from this study demonstrate that C. rodhaini and D. hominis are important causes of myiasis in owned dogs in the GAR of Ghana. Dogs could play a role in the spread of myiasis to humans, with its attendant public health implications.

  14. Economic and geographic factors affecting the development of Greater Baku

    Directory of Open Access Journals (Sweden)

    Vusat AFANDIYEV

    2014-12-01

    Full Text Available Globally, the responsible factors for the ongoing development of urbanization are the high speed of population growth, and the mass migration of humans to cities and large urban areas. In most countries, this process resulted in the emergence of ‘pseudo-urbanization’ which is difficult to be regulated. The purpose of the carried researches to determine the development priorities in the territory of Greater Baku – the capital city of the Republic of Azerbaijan; to define the problems that take place in this connection; and to develop ways of elimination of these problems. The reason of taking Baku as a research area is connected with some of the factors. Firstly, studies on Baku have been conducted based on the Soviet geographical and urban planning school and their methods for a long period. In this regard, it is necessary to carry out research in this field based on the principles adopted in most countries. Secondly, since 1992, the intensive accumulation of population in the territory of the capital city and the surrounding areas is being observed because of socio-economic problems. As a result, the process of pseudo-urbanization intensified, entailing a densely-populated area. Thirdly, low-rise buildings still continue to exist in the large areas within the territory of Baku, and they are not associated with the functional structure of the city. This situation creates many challenges, particularly in terms of density growth and effective use of the city’s territory. Finally, numerous new buildings have been constructed in the residential areas of Baku in recent years, and this may entailserious problems in water supply, energy provision, and utilities. The study is carried out referring to previous works of researchers, statistic data, and the results of the population census conducted in 1959-2009.The practical significance of the scientific work is that positive and negative factors affecting the further development of Greater Baku

  15. Greater learnability is not sufficient to produce cultural universals.

    Science.gov (United States)

    Rafferty, Anna N; Griffiths, Thomas L; Ettlinger, Marc

    2013-10-01

    Looking across human societies reveals regularities in the languages that people speak and the concepts that they use. One explanation that has been proposed for these "cultural universals" is differences in the ease with which people learn particular languages and concepts. A difference in learnability means that languages and concepts possessing a particular property are more likely to be accurately transmitted from one generation of learners to the next. Intuitively, this difference could allow languages and concepts that are more learnable to become more prevalent after multiple generations of cultural transmission. If this is the case, the prevalence of languages and concepts with particular properties can be explained simply by demonstrating empirically that they are more learnable. We evaluate this argument using mathematical analysis and behavioral experiments. Specifically, we provide two counter-examples that show how greater learnability need not result in a property becoming prevalent. First, more learnable languages and concepts can nonetheless be less likely to be produced spontaneously as a result of transmission failures. We simulated cultural transmission in the laboratory to show that this can occur for memory of distinctive items: these items are more likely to be remembered, but not generated spontaneously once they have been forgotten. Second, when there are many languages or concepts that lack the more learnable property, sheer numbers can swamp the benefit produced by greater learnability. We demonstrate this using a second series of experiments involving artificial language learning. Both of these counter-examples show that simply finding a learnability bias experimentally is not sufficient to explain why a particular property is prevalent in the languages or concepts used in human societies: explanations for cultural universals based on cultural transmission need to consider the full set of hypotheses a learner could entertain and all of

  16. Greater general startle reflex is associated with greater anxiety levels: a correlational study on 111 young women

    Directory of Open Access Journals (Sweden)

    Eleonora ePoli

    2015-02-01

    Full Text Available Startle eyeblink reflex is a valid non-invasive tool for studying attention, emotion and psychiatric disorders. In the absence of any experimental manipulation, the general (or baseline startle reflex shows a high inter-individual variability, which is often considered task-irrelevant and therefore normalized across participants. Unlike the above view, we hypothesized that greater general startle magnitude is related to participants’ higher anxiety level. 111 healthy young women, after completing the State-Trait Anxiety Inventory, were randomly administered 10 acoustic white noise probes (50 ms, 100 dBA acoustic level while integrated EMG from left and right orbicularis oculi was recorded. Results showed that participants with greater state anxiety levels exhibited larger startle reflex magnitude from the left eye (r109=0.23, p<0.05. Furthermore, individuals who perceived the acoustic probe as more aversive reported the largest anxiety scores (r109=0.28, p<0.05 and had the largest eyeblinks, especially in the left eye (r109 = 0.34, p<0.001. Results suggest that general startle may represent a valid tool for studying the neural excitability underlying anxiety and emotional dysfunction in neurological and mental disorders.

  17. Urban-rural solar radiation loss in the atmosphere of Greater Cairo region, Egypt

    International Nuclear Information System (INIS)

    Robaa, S.M.

    2009-01-01

    A comparative study for measured global solar radiation, G, during the period (1969-2006) and the corresponding global radiation loss in the atmosphere, R L %, over urban and rural districts in Greater Cairo region have been performed. The climatic variabilities of G radiation at the urban and rural sites are also investigated and discussed. Monthly, seasonal and annual mean values of extraterrestrial radiation, Go, and R L % during four successive periods, (1969-1978), (1979-1988), (1989-1998) and (1999-2006) at the above two sites have been calculated and investigated. The results revealed that urban area was always received lower amount of solar radiation due to urbanization factors. The yearly mean values of G radiation were distinctly decreased from maximum value 21.93 and 22.62 MJ m -2 during 1970 year to minimum value 17.57 and 17.87 MJ m -2 during 2004 and 2006 years with average decrease rate 0.09 and 0.10 MJ m -2 per year for the urban and rural areas, respectively. Also, the seasonal and annual mean anomalies of G radiation have been also gradually decreased from maximum values during the eldest period (1969-1978) to minimum values during the recent period (1999-2006). R L % over the urban area was always higher than that rural area. The urban-rural R L % differences range from 0.61% in 1999 year to 4.19% in 2002 year and 2.20% as average value. The yearly mean of R L % values distinctly gradually increase from minimum value 29.47% and 27.28% during 1970 year to maximum value 43.50% and 42.60% during 2004 and 2006 years with average increase rate 0.28% and 0.32% per year for the urban and rural areas, respectively. The minimum value of R L % (26.88%) occurred at rural area during summer season of the eldest period (1969-1978) while the maximum value of R L % (51.27%) occurred at the urban area during winter season of the last recent urbanized period (1999-2006). The linear trend of the yearly variations of R L % revealed that G values will reach zero

  18. Dynamic Study of Soil Erosion in Greater Khingan Forest

    OpenAIRE

    Wei Li; Wenyi Fan; Xuegang Mao; Xiaojie Wang

    2015-01-01

    Based on the amended model of RUSLE universal soil loss equation and GIS technology, combined with the natural geographical features of Great Khingan, it has conducted quantitative analysis of the factor in Soil loss equation. Uses 2000 and 2010 years TM images classification are land uses/cover type figure, we gets all factors values of space distribution in the RUSLE model, gets soil erosion volume estimates data and soil erosion strength distribution figure based on grid cell data and obta...

  19. Social values as arguments: similar is convincing

    Science.gov (United States)

    Maio, Gregory R.; Hahn, Ulrike; Frost, John-Mark; Kuppens, Toon; Rehman, Nadia; Kamble, Shanmukh

    2014-01-01

    Politicians, philosophers, and rhetors engage in co-value argumentation: appealing to one value in order to support another value (e.g., “equality leads to freedom”). Across four experiments in the United Kingdom and India, we found that the psychological relatedness of values affects the persuasiveness of the arguments that bind them. Experiment 1 found that participants were more persuaded by arguments citing values that fulfilled similar motives than by arguments citing opposing values. Experiments 2 and 3 replicated this result using a wider variety of values, while finding that the effect is stronger among people higher in need for cognition and that the effect is mediated by the greater plausibility of co-value arguments that link motivationally compatible values. Experiment 4 extended the effect to real-world arguments taken from political propaganda and replicated the mediating effect of argument plausibility. The findings highlight the importance of value relatedness in argument persuasiveness. PMID:25147529

  20. Social Values as Arguments: Similar is Convincing

    Directory of Open Access Journals (Sweden)

    Gregory R Maio

    2014-08-01

    Full Text Available Politicians, philosophers, and rhetors engage in co-value argumentation: appealing to one value in order to support another value (e.g., equality leads to freedom. Across four experiments in the United Kingdom and India, we found that the psychological relatedness of values affects the persuasiveness of the arguments that bind them. Experiment 1 found that participants were more persuaded by arguments citing values that fulfilled similar motives than by arguments citing opposing values. Experiments 2 and 3 replicated this result using a wider variety of values, while finding that the effect is stronger among people higher in need for cognition and that the effect is mediated by the greater plausibility of co-value arguments that link motivationally compatible values. Experiment 4 extended the effect to real-world arguments taken from political propaganda and replicated the mediating effect of argument plausibility. The findings highlight the importance of value relatedness in argument persuasiveness.