An in vitro assay to study the recruitment and substrate specificity of chromatin modifying enzymes

Directory of Open Access Journals (Sweden)

Vermeulen Michiel

2004-01-01

Full Text Available Post-translational modifications of core histones play an important role in regulating fundamental biological processes such as DNA repair, transcription and replication. In this paper, we describe a novel assay that allows sequential targeting of distinct histone modifying enzymes to immobilized nucleosomal templates using recombinant chimeric targeting molecules. The assay can be used to study the histone substrate specificity of chromatin modifying enzymes as well as whether and how certain enzymes affect each other's histone modifying activities. As such the assay can help to understand how a certain histone code is established and interpreted.

Simulation of dislocation glide in dilute Fe-Cu alloys

International Nuclear Information System (INIS)

Tapasa, K.; Bacon, D.J.; Osetsky, Yu.N.

2005-01-01

The effects on dislocation glide of the substitutional element copper in solution in α-iron are being investigated by computer simulation. In the first phase, the critical stress for a 1/2 {110} edge dislocation to overcome configurations of either a single or two nearest-neighbour solute atoms is simulated. Molecular statics and dynamics methods are used to simulate effects at temperature equal to and greater than 0K, respectively. Single copper atoms and nearest-neighbour pairs in the first atomic plane below the glide plane give the strongest barrier to dislocation glide, in partial agreement with elasticity theory. In addition to temperature, obstacle-spacing effects are considered

Efficiency of Lift Production in Flapping and Gliding Flight of Swifts

Science.gov (United States)

Henningsson, Per; Hedenström, Anders; Bomphrey, Richard J.

2014-01-01

Many flying animals use both flapping and gliding flight as part of their routine behaviour. These two kinematic patterns impose conflicting requirements on wing design for aerodynamic efficiency and, in the absence of extreme morphing, wings cannot be optimised for both flight modes. In gliding flight, the wing experiences uniform incident flow and the optimal shape is a high aspect ratio wing with an elliptical planform. In flapping flight, on the other hand, the wing tip travels faster than the root, creating a spanwise velocity gradient. To compensate, the optimal wing shape should taper towards the tip (reducing the local chord) and/or twist from root to tip (reducing local angle of attack). We hypothesised that, if a bird is limited in its ability to morph its wings and adapt its wing shape to suit both flight modes, then a preference towards flapping flight optimization will be expected since this is the most energetically demanding flight mode. We tested this by studying a well-known flap-gliding species, the common swift, by measuring the wakes generated by two birds, one in gliding and one in flapping flight in a wind tunnel. We calculated span efficiency, the efficiency of lift production, and found that the flapping swift had consistently higher span efficiency than the gliding swift. This supports our hypothesis and suggests that even though swifts have been shown previously to increase their lift-to-drag ratio substantially when gliding, the wing morphology is tuned to be more aerodynamically efficient in generating lift during flapping. Since body drag can be assumed to be similar for both flapping and gliding, it follows that the higher total drag in flapping flight compared with gliding flight is primarily a consequence of an increase in wing profile drag due to the flapping motion, exceeding the reduction in induced drag. PMID:24587260

Efficiency of lift production in flapping and gliding flight of swifts.

Directory of Open Access Journals (Sweden)

Per Henningsson

Full Text Available Many flying animals use both flapping and gliding flight as part of their routine behaviour. These two kinematic patterns impose conflicting requirements on wing design for aerodynamic efficiency and, in the absence of extreme morphing, wings cannot be optimised for both flight modes. In gliding flight, the wing experiences uniform incident flow and the optimal shape is a high aspect ratio wing with an elliptical planform. In flapping flight, on the other hand, the wing tip travels faster than the root, creating a spanwise velocity gradient. To compensate, the optimal wing shape should taper towards the tip (reducing the local chord and/or twist from root to tip (reducing local angle of attack. We hypothesised that, if a bird is limited in its ability to morph its wings and adapt its wing shape to suit both flight modes, then a preference towards flapping flight optimization will be expected since this is the most energetically demanding flight mode. We tested this by studying a well-known flap-gliding species, the common swift, by measuring the wakes generated by two birds, one in gliding and one in flapping flight in a wind tunnel. We calculated span efficiency, the efficiency of lift production, and found that the flapping swift had consistently higher span efficiency than the gliding swift. This supports our hypothesis and suggests that even though swifts have been shown previously to increase their lift-to-drag ratio substantially when gliding, the wing morphology is tuned to be more aerodynamically efficient in generating lift during flapping. Since body drag can be assumed to be similar for both flapping and gliding, it follows that the higher total drag in flapping flight compared with gliding flight is primarily a consequence of an increase in wing profile drag due to the flapping motion, exceeding the reduction in induced drag.

The glide of screw dislocations in bcc Fe: Atomistic static and dynamic simulations

International Nuclear Information System (INIS)

Chaussidon, Julien; Fivel, Marc; Rodney, David

2006-01-01

We present atomic-scale simulations of screw dislocation glide in bcc iron. Using two interatomic potentials that, respectively, predict degenerate and non-degenerate core structures, we compute the static 0 K dependence of the screw dislocation Peierls stress on crystal orientation and show strong boundary condition effects related to the generation of non-glide stress components. At finite temperatures we show that, with a non-degenerate core, glide by nucleation/propagation of kink-pairs in a {1 1 0} glide plane is obtained at low temperatures. A transition in the twinning region, towards an average {1 1 2} glide plane, with the formation of debris loops is observed at higher temperatures

Cytochrome P450-mediated metabolism of tumour promoters modifies the inhibition of intercellular communication: a modified assay for tumour promotion

DEFF Research Database (Denmark)

Vang, Ole; Wallin, H.; Doehmer, J.

1993-01-01

The role of metabolism of tumour promoters on the inhibition of intercellular communication was investigated in a modified V79 metabolic cooperation system. V79 cells, which stably express different rat cytochrome P450 enzymes (CYP1A1, CYP1A2 or CYP2B1), were used in the metabolic cooperation assay...... B1 and 4-nitrobiphenyl, did not inhibit metabolic cooperation in either V79 cells expressing or cells not expressing cytochrome P450. We conclude that cytochrome P450-associated metabolism plays an important role in the inhibition of gap junctional intercellular communication of some tumour...... promoters. The modified metabolic cooperation assay presented here is valuable for detecting some inhibitory chemicals which have been 'false negative' in previous assays for gap junctional intercellular communication. The assay also discloses that cytochrome P450 metabolism alters intercellular...

Onboard Determination of Vehicle Glide Capability for Shuttle Abort Flight Managment (SAFM)

Science.gov (United States)

Straube, Timothy; Jackson, Mark; Fill, Thomas; Nemeth, Scott

2002-01-01

When one or more main engines fail during ascent, the flight crew of the Space Shuttle must make several critical decisions and accurately perform a series of abort procedures. One of the most important decisions for many aborts is the selection ofa landing site. Several factors influence the ability to reach a landing site, including the spacecraft point of atmospheric entry, the energy state at atmospheric entry, the vehicle glide capability from that energy state, and whether one or more suitable landing sites are within the glide capability. Energy assessment is further complicated by the fact that phugoid oscillations in total energy influence glide capability. Once the glide capability is known, the crew must select the "best" site option based upon glide capability and landing site conditions and facilities. Since most of these factors cannot currently be assessed by the crew in flight, extensive planning is required prior to each mission to script a variety of procedures based upon spacecraft velocity at the point of engine failure (or failures). The results of this preflight planning are expressed in tables and diagrams on mission-specific cockpit checklists. Crew checklist procedures involve leafing through several pages of instructions and navigating a decision tree for site selection and flight procedures - all during a time critical abort situation. With the advent of the Cockpit Avionics Upgrade (CAU), the Shuttle will have increased on-board computational power to help alleviate crew workload during aborts and provide valuable situational awareness during nominal operations. One application baselined for the CAU computers is Shuttle Abort Flight Management (SAFM), whose requirements have been designed and prototyped. The SAFM application includes powered and glided flight algorithms. This paper describes the glided flight algorithm which is dispatched by SAFM to determine the vehicle glide capability and make recommendations to the crew for site

Influence of glide path on the screw-in effect and torque of nickel-titanium rotary files in simulated resin root canals

Directory of Open Access Journals (Sweden)

Jung-Hong Ha

2012-11-01

Full Text Available Objectives The purpose of this study was to investigate the screw-in effect and torque generation depending on the size of glide path during root canal preparation. Materials and Methods Forty Endo-Training Blocks (REF A 0177, Dentsply Maillefer were used. They were divided into 4 groups. For groups 1, 2, 3, and 4, the glide path was established with ISO #13 Path File (Dentsply Maillefer, #15 NiTi K-file NITIFLEX (Dentsply Maillefer, modified #16 Path File (equivalent to #18, and #20 NiTi K-file NITIFLEX, respectively. The screw-in force and resultant torque were measured using a custom-made experimental apparatus while canals were instrumented with ProTaper S1 (Dentsply Maillefer at a constant speed of 300 rpm with an automated pecking motion. A statistical analysis was performed using one-way analysis of variance and the Duncan post hoc comparison test. Results Group 4 showed lowest screw-in effect (2.796 ± 0.134 among the groups (p < 0.05. Torque was inversely proportional to the glide path of each group. In #20 glide path group, the screw-in effect and torque decreased at the last 1 mm from the apical terminus. However, in the other groups, the decrease of the screw-in effect and torque did not occur in the last 1 mm from the apical terminus. Conclusions The establishment of a larger glide path before NiTi rotary instrumentation appears to be appropriate for safely shaping the canal. It is recommended to establish #20 glide path with NiTi file when using ProTaper NiTi rotary instruments system safely.

Debris extrusion by glide-path establishing endodontic instruments with different geometries

Directory of Open Access Journals (Sweden)

Jung-Hong Ha

2016-06-01

Conclusion: Creating the glide-path using nickel-titanium rotary files produced lower amounts of debris extrusion than using manual stainless-steel files. The progressive taper design of ProGlider, the center-off cross-section of One G, and the alternative-pitch design of ScoutRace may have increased the efficiencies of debris removal with minimal extrusion during glide-path preparation. Glide-path preparation using NiTi rotary files have better clinical efficiency than the manual stainless-steel file.

Director gliding in a nematic liquid crystal layer: Quantitative comparison with experiments

Science.gov (United States)

Mema, E.; Kondic, L.; Cummings, L. J.

2018-03-01

The interaction between nematic liquid crystals and polymer-coated substrates may lead to slow reorientation of the easy axis (so-called "director gliding") when a prolonged external field is applied. We consider the experimental evidence of zenithal gliding observed by Joly et al. [Phys. Rev. E 70, 050701 (2004), 10.1103/PhysRevE.70.050701] and Buluy et al. [J. Soc. Inf. Disp. 14, 603 (2006), 10.1889/1.2235686] as well as azimuthal gliding observed by S. Faetti and P. Marianelli [Liq. Cryst. 33, 327 (2006), 10.1080/02678290500512227], and we present a simple, physically motivated model that captures the slow dynamics of gliding, both in the presence of an electric field and after the electric field is turned off. We make a quantitative comparison of our model results and the experimental data and conclude that our model explains the gliding evolution very well.

14 CFR 29.71 - Helicopter angle of glide: Category B.

Science.gov (United States)

2010-01-01

... 14 Aeronautics and Space 1 2010-01-01 2010-01-01 false Helicopter angle of glide: Category B. 29... AIRCRAFT AIRWORTHINESS STANDARDS: TRANSPORT CATEGORY ROTORCRAFT Flight Performance § 29.71 Helicopter angle of glide: Category B. For each category B helicopter, except multiengine helicopters meeting the...

Translational, rotational, vibrational and electron temperatures of a gliding arc discharge

DEFF Research Database (Denmark)

Zhu, Jiajian; Ehn, Andreas; Gao, Jinlong

2017-01-01

, 0) band was used to simulate the rotational temperature (Tr) of the gliding arc discharge whereas the NO A–X (1, 0) and (0, 1) bands were used to determine its vibrational temperature (Tv). The instantaneous reduced electric field strength E/N was obtained by simultaneously measuring......Translational, rotational, vibrational and electron temperatures of a gliding arc discharge in atmospheric pressure air were experimentally investigated using in situ, non-intrusive optical diagnostic techniques. The gliding arc discharge was driven by a 35 kHz alternating current (AC) power source...... and operated in a glow-type regime. The two-dimensional distribution of the translational temperature (Tt) of the gliding arc discharge was determined using planar laser-induced Rayleigh scattering. The rotational and vibrational temperatures were obtained by simulating the experimental spectra. The OH A–X (0...

Non-equilibrium trajectory dynamics and the kinematics of gliding in a flying snake

International Nuclear Information System (INIS)

Socha, John J; Jafari, Farid; Miklasz, Kevin; Vlachos, Pavlos P

2010-01-01

Given sufficient space, it is possible for gliding animals to reach an equilibrium state with no net forces acting on the body. In contrast, every gliding trajectory must begin with a non-steady component, and the relative importance of this phase is not well understood. Of any terrestrial animal glider, snakes exhibit the greatest active movements, which may affect their trajectory dynamics. Our primary aim was to determine the characteristics of snake gliding during the transition to equilibrium, quantifying changes in velocity, acceleration, and body orientation in the late phase of a glide sequence. We launched 'flying' snakes (Chrysopelea paradisi) from a 15 m tower and recorded the mid-to-end portion of trajectories with four videocameras to reconstruct the snake's body position with mm to cm accuracy. Additionally, we developed a simple analytical model of gliding assuming only steady-state forces of lift, drag and weight acting on the body and used it to explore effects of wing loading, lift-to-drag ratio, and initial velocity on trajectory dynamics. Despite the vertical space provided to transition to steady-state gliding, snakes did not exhibit equilibrium gliding and in fact displayed a net positive acceleration in the vertical axis, an effect also predicted by the analytical model.

Non-equilibrium trajectory dynamics and the kinematics of gliding in a flying snake

Energy Technology Data Exchange (ETDEWEB)

Socha, John J; Jafari, Farid [Engineering Science and Mechanics, Virginia Tech, Blacksburg, VA 24061 (United States); Miklasz, Kevin [Hopkins Marine Station, Stanford University, Pacific Grove, CA 93950 (United States); Vlachos, Pavlos P, E-mail: jjsocha@vt.ed [Department of Mechanical Engineering, Virginia Tech, Blacksburg, VA 24061 (United States)

2010-12-15

Given sufficient space, it is possible for gliding animals to reach an equilibrium state with no net forces acting on the body. In contrast, every gliding trajectory must begin with a non-steady component, and the relative importance of this phase is not well understood. Of any terrestrial animal glider, snakes exhibit the greatest active movements, which may affect their trajectory dynamics. Our primary aim was to determine the characteristics of snake gliding during the transition to equilibrium, quantifying changes in velocity, acceleration, and body orientation in the late phase of a glide sequence. We launched 'flying' snakes (Chrysopelea paradisi) from a 15 m tower and recorded the mid-to-end portion of trajectories with four videocameras to reconstruct the snake's body position with mm to cm accuracy. Additionally, we developed a simple analytical model of gliding assuming only steady-state forces of lift, drag and weight acting on the body and used it to explore effects of wing loading, lift-to-drag ratio, and initial velocity on trajectory dynamics. Despite the vertical space provided to transition to steady-state gliding, snakes did not exhibit equilibrium gliding and in fact displayed a net positive acceleration in the vertical axis, an effect also predicted by the analytical model.

Assessing cellulolysis in passive treatment systems for mine drainage: a modified enzyme assay.

Science.gov (United States)

McDonald, Corina M; Gould, W Douglas; Lindsay, Matthew B J; Blowes, David W; Ptacek, Carol J; Condon, Peter D

2013-01-01

A modified cellulase enzyme assay was developed to monitor organic matter degradation in passive treatment systems for mine drainage. This fluorogenic substrate method facilitates assessment of exo-(1,4)-β-D-glucanase, endo-(1,4)-β-D-glucanase, and β-glucosidase, which compose an important cellulase enzyme system. The modified method was developed and refined using samples of organic carbon-amended mine tailings from field experiments where sulfate reduction was induced as a strategy for managing water quality. Sample masses (3 g) and the number of replicates ( ≥ 3) were optimized. Matrix interferences within these metal-rich samples were found to be insignificant. Application of this modified cellulase assay method provided insight into the availability and degradation of organic carbon within the amended tailings. Results of this study indicate that cellulase enzyme assays can be applied to passive treatment systems for mine drainage, which commonly contain elevated concentrations of metals. Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

Lubrication of dislocation glide in MgO by hydrous defects

Science.gov (United States)

Skelton, Richard; Walker, Andrew M.

2018-02-01

Water-related defects, principally in the form of protonated cation vacancies, are potentially able to weaken minerals under high-stress or low-temperature conditions by reducing the Peierls stress required to initiate dislocation glide. In this study, we use the Peierls-Nabarro (PN) model to determine the effect of protonated Mg vacancies on the 1/2{110} and 1/2{100} slip systems in MgO. This PN model is parameterized using generalized stacking fault energies calculated using plane-wave density functional theory, with and without protonated Mg vacancies present at the glide plane. It found that these defects increase dislocation core widths and reduce the Peierls stress over the entire pressure range 0-125 GPa. Furthermore, 1/2{110} slip is found to be more sensitive to the presence of protonated vacancies which increases in the pressure at which {100} becomes the easy glide plane for 1/2 screw dislocations. These results demonstrate, for a simple mineral system, that water-related defects can alter the deformation behavior of minerals in the glide-creep regime by reducing the stress required to move dislocations by glide. (Mg, Fe)O is the most anisotropic mineral in the Earth's lower mantle, so the differential sensitivity of the major slip systems in MgO to hydrous defects has potential implications for the interpretation of the seismic anisotropy in this region.

Decomposition of naphthalene by dc gliding arc gas discharge.

Science.gov (United States)

Yu, Liang; Li, Xiaodong; Tu, Xin; Wang, Yu; Lu, Shengyong; Yan, Jianhua

2010-01-14

Gliding arc discharge has been proved to be effective in treatment of gas and liquid contaminants. In this study, physical characteristics of dc gliding arc discharge and its application to naphthalene destruction are investigated with different external resistances and carrier gases. The decomposition rate increases with increasing of oxygen concentration and decreases with external resistance. This value can be achieved up to 92.3% at the external resistance of 50 kOmega in the oxygen discharge, while the highest destruction energy efficiency reaches 3.6 g (kW h)(-1) with the external resistance of 93 kOmega. Possible reaction pathways and degradation mechanisms in the plasma with different gases are proposed by qualitative analysis of postdestructed products. In the air and oxygen gliding arc discharges, the naphthalene degradation is mainly governed by reactions with oxygen-derived radicals.

The novel echo-guided ProGlide technique during percutaneous transfemoral transcatheter aortic valve implantation.

Science.gov (United States)

Honda, Yohsuke; Araki, Motoharu; Yamawaki, Masahiro; Tokuda, Takahiro; Tsutumi, Masakazu; Mori, Shinsuke; Sakamoto, Yasunari; Kobayashi, Norihiro; Hirano, Keisuke; Ito, Yoshiaki

2018-04-01

The aim of this study was to assess clinical benefit of the Echo-guided ProGlide technique in patients undergoing percutaneous transfemoral transcatheter aortic valve implantation (TF-TAVI). The efficacy of the Echo-guided ProGlide technique during percutaneous TF-TAVI was not previously clarified. A total of 121 consecutive patients who underwent percutaneous TF-TAVI at our institution between February 2014 and July 2017 were enrolled in this study. According to the introduction of this novel technique in March 2016, patients were divided into two groups (echo-guided group who underwent TAVI from March 2016 to July 2017, n = 63; not echo-guided group who underwent TAVI from February 2014 to February 2016, n = 58). The incidence of major vascular complications, defined per the Valve Academic Research Consortium-2 criteria, and ProGlide complications including acute femoral artery stenosis or occlusion and bleeding requiring any intervention. The incidence of major vascular complication and ProGlide complication were significantly lower in the echo-guided group than in not echo-guided group (1.6% vs 17.2%, P guided ProGlide technique was independently associated with prevention of ProGlide complications (odds ratio, 0.11; 95% confidential interval, 0.01-0.76; P = 0.03). This novel Echo-guided ProGlide technique was associated with a lower rate of major vascular complications, particularly ProGlide complications during percutaneous TF-TAVI. © 2017, Wiley Periodicals, Inc.

The modified high-density survival assay is the useful tool to predict the effectiveness of fractionated radiation exposure

International Nuclear Information System (INIS)

Kuwahara, Yoshikazu; Mori, Miyuki; Oikawa, Toshiyuki; Shimura, Tsutomu; Fukumoto, Manabu; Ohtake, Yosuke; Ohkubo, Yasuhito; Mori, Shiro

2010-01-01

The high-density survival (HDS) assay was originally elaborated to assess cancer cell responses to therapeutic agents under the influence of intercellular communication. Here, we simplified the original HDS assay and studied its applicability for the detection of cellular radioresistance. We have recently defined clinically relevant radioresistant (CRR) cells, which continue to proliferate with daily exposure to 2 gray (Gy) of X-rays for more than 30 days in vitro. We established human CRR cell lines, HepG2-8960-R from HepG2, and SAS-R1 and -R2 from SAS, respectively. In an attempt to apply the HDS assay to detect radioresistance with clinical relevance, we simplified the original HDS assay by scoring the total number of surviving cells after exposure to X-rays. The modified HDS assay successfully detected radioresistance with clinical relevance. The modified HDS assay detected CRR phenotype, which is not always detectable by clonogenic assay. Therefore, we believe that the modified HDS assay presented in this study is a powerful tool to predict the effectiveness of fractionated radiotherapy against malignant tumors. (author)

Plasma-liquid system with rotational gliding discharge with liquid electrode

International Nuclear Information System (INIS)

Nedybaliuk, O.A.; Solomenko, O.V; Martysh, E.V.; Fedirchuk, I.I.

2014-01-01

Plasma-liquid system based on rotational gliding discharge with one liquid electrode was developed. Emission spectra of plasma of rotational gliding discharge with one liquid electrode were investigated. Discovered effective mechanism of controlling non-isothermal level of plasma in dynamic plasma-liquid systems. Major mechanism of expulsion of metal anode material from plasma-liquid systems with rotational discharges was shown.

Evolution of gliding in Southeast Asian geckos and other vertebrates is temporally congruent with dipterocarp forest development.

Science.gov (United States)

Heinicke, Matthew P; Greenbaum, Eli; Jackman, Todd R; Bauer, Aaron M

2012-12-23

Gliding morphologies occur in diverse vertebrate lineages in Southeast Asian rainforests, including three gecko genera, plus frogs, snakes, agamid lizards and squirrels. It has been hypothesized that repeated evolution of gliding is related to the dominance of Asian rainforest tree floras by dipterocarps. For dipterocarps to have influenced the evolution of gliding in Southeast Asian vertebrates, gliding lineages must have Eocene or later origins. However, divergence times are not known for most lineages. To investigate the temporal pattern of Asian gliding vertebrate evolution, we performed phylogenetic and molecular clock analyses. New sequence data for geckos incorporate exemplars of each gliding genus (Cosymbotus, Luperosaurus and Ptychozoon), whereas analyses of other vertebrate lineages use existing sequence data. Stem ages of most gliding vertebrates, including all geckos, cluster in the time period when dipterocarps came to dominate Asian tropical forests. These results demonstrate that a gliding/dipterocarp correlation is temporally viable, and caution against the assumption of early origins for apomorphic taxa.

Use of glide-ins in CMS for production and analysis

International Nuclear Information System (INIS)

Bradley, D; Gutsche, O; Holzman, B; Sfiligoi, I; Vaandering, E; Hahn, K; Padhi, S; Pi, H; Wuerthwein, F; Spiga, D

2010-01-01

With the evolution of various grid federations, the Condor glide-ins represent a key feature in providing a homogeneous pool of resources using late-binding technology. The CMS collaboration uses the glide-in based Workload Management System, glideinWMS, for production (ProdAgent) and distributed analysis (CRAB) of the data. The Condor glide-in daemons traverse to the worker nodes, submitted via Condor-G. Once activated, they preserve the Master-Worker relationships, with the worker first validating the execution environment on the worker node before pulling the jobs sequentially until the expiry of their lifetimes. The combination of late-binding and validation significantly reduces the overall failure rate visible to CMS physicists. We discuss the extensive use of the glideinWMS since the computing challenge, CCRC-08, in order to prepare for the forthcoming LHC data-taking period. The key features essential to the success of large-scale production and analysis on CMS resources across major grid federations, including EGEE, OSG and NorduGrid are outlined. Use of glide-ins via the CRAB server mechanism and ProdAgent, as well as first hand experience of using the next generation CREAM computing element within the CMS framework is discussed.

Gliding swifts attain laminar flow over rough wings.

Directory of Open Access Journals (Sweden)

David Lentink

Full Text Available Swifts are among the most aerodynamically refined gliding birds. However, the overlapping vanes and protruding shafts of their primary feathers make swift wings remarkably rough for their size. Wing roughness height is 1-2% of chord length on the upper surface--10,000 times rougher than sailplane wings. Sailplanes depend on extreme wing smoothness to increase the area of laminar flow on the wing surface and minimize drag for extended glides. To understand why the swift does not rely on smooth wings, we used a stethoscope to map laminar flow over preserved wings in a low-turbulence wind tunnel. By combining laminar area, lift, and drag measurements, we show that average area of laminar flow on swift wings is 69% (n = 3; std 13% of their total area during glides that maximize flight distance and duration--similar to high-performance sailplanes. Our aerodynamic analysis indicates that swifts attain laminar flow over their rough wings because their wing size is comparable to the distance the air travels (after a roughness-induced perturbation before it transitions from laminar to turbulent. To interpret the function of swift wing roughness, we simulated its effect on smooth model wings using physical models. This manipulation shows that laminar flow is reduced and drag increased at high speeds. At the speeds at which swifts cruise, however, swift-like roughness prolongs laminar flow and reduces drag. This feature gives small birds with rudimentary wings an edge during the evolution of glide performance.

[Posterior lamellar keratoplasty with DSEK technique and use of the Tan EndoGlide - short-term results].

Science.gov (United States)

Kałuiny, Bartłomiej J; Piotrowiak, Ilona; Sołdańska, Beata; Grzybek, Katarzyna; Czajkowska, Monika; Galas, Małgorzata; Malukiewicz, Grazyna

2013-01-01

To present the differences in surgical technique of DSEK (Descemet's Stripping Endothelial Keratoplasty) with the use of Tan EndoGlide (Coronet, UK) and Busin Glide (Moria, FR). Short-term results will also be presented, DSEK was performed in 24 eyes, in 8 cases the surgery was combined with cataract phacoemulsification and lOL implantation. Surgery course and 6 months postoperative results of first 12 eyes performed with the use of Tan EndoGlide were compared with 12 consecutive eyes preformed with Busin Glide. Tan EndoGlide provided much more stable anterior chamber, donor tissue unfolding process was better controlled but the incision was wider incision. Surgically induced mean refractory cylinder 6. months after the surgery was 1.56 - 1.15 Dsph in Tan EndoGlide group and 1.18 +/- 1.10 Dsph in Busin Glide group (P 0.05). Mean CDVA was 0.65+/- 0.27 and 0.63 +/- 0.25, respectively (P>0,05). Statistically significant differences in intra- and post-operative complications between both groups were not found. The Tan EndoGlide used during posterior lamellar keratoplasty with DSEK technique is a good alternative to currently used methods. It provides better stabilization of the anterior chamber, however its use is linked with higher postoperative astigmatism in comparison with Busin Glide. The visual outcomes and endothelial cell loss 6 months after the surgery were similar in both groups.

Improving efficiency of heat pumps by use of zeotropic mixtures for different temperature glides

DEFF Research Database (Denmark)

Zühlsdorf, Benjamin; Jensen, Jonas Kjær; Cignitti, Stefano

2017-01-01

The present study demonstrates the optimization of a heat pump for an application with a large temperature glide on the sink and a smaller temperature glide on the source side. The study includes a simulation of a heat pump cycle for all possible binary mixtures from a list of 14 natural...... refrigerants, which enables a match of the temperature glide of sink and source with the temperature of the working fluid during phase change and thus, a reduction of the exergy destruction due to heat transfer. The model was evaluated for four different boundary conditions. For a separated evaluation...... of the irreversibility solely caused by the fluid properties, the exergy destruction in the heat exchangers has been distinguished accordingly and an indicator quantifying the glide match has been defined to analyse the influence on the performance. It was observed that a good glide match can contribute to an increased...

Plastic deformation of crystals: analytical and computer simulation studies of dislocation glide

International Nuclear Information System (INIS)

Altintas, S.

1978-05-01

The plastic deformation of crystals is usually accomplished through the motion of dislocations. The glide of a dislocation is impelled by the applied stress and opposed by microstructural defects such as point defects, voids, precipitates and other dislocations. The planar glide of a dislocation through randomly distributed obstacles is considered. The objective of the present research work is to calculate the critical resolved shear stress (CRSS) for athermal glide and the velocity of the dislocation at finite temperature as a function of the applied stress and the nature and strength of the obstacles. Dislocation glide through mixtures of obstacles has been studied analytically and by computer simulation. Arrays containing two kinds of obstacles as well as square distribution of obstacle strengths are considered. The critical resolved shear stress for an array containing obstacles with a given distribution of strengths is calculated using the sum of the quadratic mean of the stresses for the individual obstacles and is found to be in good agreement with the computer simulation data. Computer simulation of dislocation glide through randomly distributed obstacles containing up to 10 6 obstacles show that the CRSS decreases as the size of the array increases and approaches a limiting value. Histograms of forces and of segment lengths are obtained and compared with theoretical predictions. Effects of array shape and boundary conditions on the dislocation glide are also studied. Analytical and computer simulation results are compared with experimental results obtained on precipitation-, irradiation-, forest-, and impurity cluster-hardening systems and are found to be in good agreement

Plastic deformation of crystals: analytical and computer simulation studies of dislocation glide

Energy Technology Data Exchange (ETDEWEB)

Altintas, S.

1978-05-01

The plastic deformation of crystals is usually accomplished through the motion of dislocations. The glide of a dislocation is impelled by the applied stress and opposed by microstructural defects such as point defects, voids, precipitates and other dislocations. The planar glide of a dislocation through randomly distributed obstacles is considered. The objective of the present research work is to calculate the critical resolved shear stress (CRSS) for athermal glide and the velocity of the dislocation at finite temperature as a function of the applied stress and the nature and strength of the obstacles. Dislocation glide through mixtures of obstacles has been studied analytically and by computer simulation. Arrays containing two kinds of obstacles as well as square distribution of obstacle strengths are considered. The critical resolved shear stress for an array containing obstacles with a given distribution of strengths is calculated using the sum of the quadratic mean of the stresses for the individual obstacles and is found to be in good agreement with the computer simulation data. Computer simulation of dislocation glide through randomly distributed obstacles containing up to 10/sup 6/ obstacles show that the CRSS decreases as the size of the array increases and approaches a limiting value. Histograms of forces and of segment lengths are obtained and compared with theoretical predictions. Effects of array shape and boundary conditions on the dislocation glide are also studied. Analytical and computer simulation results are compared with experimental results obtained on precipitation-, irradiation-, forest-, and impurity cluster-hardening systems and are found to be in good agreement.

Observation of gliding arc surface treatment

DEFF Research Database (Denmark)

Kusano, Yukihiro; Zhu, Jiajian; Ehn, A.

2015-01-01

. Water contact angle measurements indicate that the treatment uniformity improves significantly when the AC gliding arc is tilted to the polymer surface. Thickness reduction of the gas boundary layer, explaining the improvement of surface treatment, by the ultrasonic irradiation was directly observed...

Ubiquity of quantum zero-point fluctuations in dislocation glide

Science.gov (United States)

Landeiro Dos Reis, Marie; Choudhury, Anshuman; Proville, Laurent

2017-03-01

Modeling the dislocation glide through atomic scale simulations in Al, Cu, and Ni and in solid solution alloys Al(Mg) and Cu(Ag), we show that in the course of the plastic deformation the variation of the crystal zero-point energy (ZPE) and the dislocation potential energy barriers are of opposite sign. The multiplicity of situations where we have observed the same trend allows us to conclude that quantum fluctuations, giving rise to the crystal ZPE, make easier the dislocation glide in most materials, even those constituted of atoms heavier than H and He.

Identification of total reversible cysteine oxidation in an atherosclerosis model using a modified biotin switch assay.

Science.gov (United States)

Li, Ru; Huang, Jiqing; Kast, Juergen

2015-05-01

Oxidative stress due to the imbalance of reactive oxygen species (ROS) and the resulting reversible cysteine oxidation (CysOX) are involved in the early proatherogenic aspect of atherosclerosis. Given that the corresponding redox signaling pathways are still unclear, a modified biotin switch assay was developed to quantify the reversible CysOX in an atherosclerosis model established by using a monocytic cell line treated with platelet releasate. The accumulation of ROS was observed in the model system and validated in human primary monocytes. Through the application of the modified biotin switch assay, we obtained the first reversible CysOX proteome for this model. A total of 75 peptides, corresponding to 53 proteins, were quantified with oxidative modification. The bioinformatics analysis of these CysOX-containing proteins highlighted biological processes including glycolysis, cytoskeleton arrangement, and redox regulation. Moreover, the reversible oxidation of three glycolysis enzymes was observed using this method, and the regulation influence was verified by an enzyme activity assay. NADPH oxidase (NOX) inhibition treatment, in conjunction with the modified biotin switch method, was used to evaluate the global CysOX status. In conclusion, this versatile modified biotin switch assay provides an approach for the quantification of all reversible CysOX and for the study of redox signaling in atherosclerosis as well as in diseases in other biological systems.

[Endonuclease modified comet assay for oxidative DNA damage induced by detection of genetic toxicants].

Science.gov (United States)

Zhao, Jian; Li, Hongli; Zhai, Qingfeng; Qiu, Yugang; Niu, Yong; Dai, Yufei; Zheng, Yuxin; Duan, Huawei

2014-03-01

The aim of this study was to investigate the use of the lesion-specific endonucleases-modified comet assay for analysis of DNA oxidation in cell lines. DNA breaks and oxidative damage were evaluated by normal alkaline and formamidopyrimidine-DNA-glycosylase (FPG) modified comet assays. Cytotoxicity were assessed by MTT method. The human bronchial epithelial cell (16HBE) were treated with benzo (a) pyrene (B(a)P), methyl methanesulfonate (MMS), colchicine (COL) and vincristine (VCR) respectively, and the dose is 20 µmol/L, 25 mg/ml, 5 mg/L and 0.5 mg/L for 24 h, respectively. Oxidative damage was also detected by levels of reactive oxygen species in treated cells. Four genotoxicants give higher cytotoxicity and no significant changes on parameters of comet assay treated by enzyme buffer. Cell survival rate were (59.69 ± 2.60) %, (54.33 ± 2.81) %, (53.11 ± 4.00) %, (51.43 ± 3.92) % in four groups, respectively. There was the direct DNA damage induced by test genotoxicants presented by tail length, Olive tail moment (TM) and tail DNA (%) in the comet assay. The presence of FPG in the assays increased DNA migration in treated groups when compared to those without it, and the difference was statistically significant which indicated that the clastogen and aneugen could induce oxidative damage in DNA strand. In the three parameters, the Olive TM was changed most obviously after genotoxicants treatment. In the contrast group, the Olive TM of B(a) P,MMS, COL,VCR in the contrast groups were 22.99 ± 17.33, 31.65 ± 18.86, 19.86 ± 9.56 and 17.02 ± 9.39, respectively, after dealing with the FPG, the Olive TM were 34.50 ± 17.29, 43.80 ± 10.06, 33.10 ± 12.38, 28.60 ± 10.53, increased by 58.94%, 38.48%, 66.86% and 68.21%, respectively (t value was 3.91, 3.89, 6.66 and 3.87, respectively, and all P comet assay appears more specific for detecting oxidative DNA damage induced by genotoxicants exposure, and the application of comet assay will be expanded. The endonuclease

The impact of glide phases on the trackability of hydrodynamic trails in harbour seals (Phoca vitulina).

Science.gov (United States)

Wieskotten, S; Dehnhardt, G; Mauck, B; Miersch, L; Hanke, W

2010-11-01

The mystacial vibrissae of harbour seals (Phoca vitulina) constitute a highly sensitive hydrodynamic receptor system enabling the seals to detect and follow hydrodynamic trails. In the wild, hydrodynamic trails, as generated by swimming fish, consist of cyclic burst-and-glide phases, associated with various differences in the physical parameters of the trail. Here, we investigated the impact of glide phases on the trackability of differently aged hydrodynamic trails in a harbour seal. As fish are not easily trained to swim certain paths with predetermined burst-and-glide phases, the respective hydrodynamic trails were generated using a remote-controlled miniature submarine. Gliding phases in hydrodynamic trails had a negative impact on the trackability when trails were 15 s old. The seal lost the generated trails more often within the transition zones, when the submarine switched from a burst to a glide moving pattern. Hydrodynamic parameter analysis (particle image velocimetry) revealed that the smaller dimensions and faster decay of hydrodynamic trails generated by the gliding submarine are responsible for the impaired success of the seal tracking the gliding phase. Furthermore, the change of gross water flow generated by the submarine from a rearwards-directed stream in the burst phase to a water flow passively dragged behind the submarine during gliding might influence the ability of the seal to follow the trail as this might cause a weaker deflection of the vibrissae. The possible ecological implications of intermittent swimming behaviour in fish for piscivorous predators are discussed.

Analysis of temperature glide matching of heat pumps with zeotropic working fluid mixtures for different temperature glides

DEFF Research Database (Denmark)

Zühlsdorf, Benjamin; Jensen, Jonas Kjær; Cignitti, Stefano

2018-01-01

refrigerants. This approach enables a match of the temperature glide of sink and source with the temperature of the working fluid during phase change and thus, a reduction of the exergy destruction due to heat transfer. The model was evaluated for four different boundary conditions. The exergy destruction due...

Aerodynamic consequences of wing morphing during emulated take-off and gliding in birds.

Science.gov (United States)

Klaassen van Oorschot, Brett; Mistick, Emily A; Tobalske, Bret W

2016-10-01

Birds morph their wings during a single wingbeat, across flight speeds and among flight modes. Such morphing may allow them to maximize aerodynamic performance, but this assumption remains largely untested. We tested the aerodynamic performance of swept and extended wing postures of 13 raptor species in three families (Accipitridae, Falconidae and Strigidae) using a propeller model to emulate mid-downstroke of flapping during take-off and a wind tunnel to emulate gliding. Based on previous research, we hypothesized that (1) during flapping, wing posture would not affect maximum ratios of vertical and horizontal force coefficients (C V :C H ), and that (2) extended wings would have higher maximum C V :C H when gliding. Contrary to each hypothesis, during flapping, extended wings had, on average, 31% higher maximum C V :C H ratios and 23% higher C V than swept wings across all biologically relevant attack angles (α), and, during gliding, maximum C V :C H ratios were similar for the two postures. Swept wings had 11% higher C V than extended wings in gliding flight, suggesting flow conditions around these flexed raptor wings may be different from those in previous studies of swifts (Apodidae). Phylogenetic affiliation was a poor predictor of wing performance, due in part to high intrafamilial variation. Mass was only significantly correlated with extended wing performance during gliding. We conclude that wing shape has a greater effect on force per unit wing area during flapping at low advance ratio, such as take-off, than during gliding. © 2016. Published by The Company of Biologists Ltd.

Effects of Glide Path on the Centering Ability and Preparation Time of Two Reciprocating Instruments

Science.gov (United States)

Coelho, Marcelo Santos; Fontana, Carlos Eduardo; Kato, Augusto Shoji; de Martin, Alexandre Sigrist; da Silveira Bueno, Carlos Eduardo

2016-01-01

Introduction: The aim of this in vitro study was to evaluate the effects of establishing glide path on the centering ability and preparation time of two single-file reciprocating systems in mesial root canals of mandibular molars. Methods and Materials: Sixty extracted mandibular molars with curvatures of 25-39 degrees and separate foramina for the mesiobuccal and mesiolingual canals, were divided into four groups (n=15); WaveOne+glide path; WaveOne; Reciproc+glide path and Reciproc. Non-patent canals were excluded and only one canal in each tooth was instrumented. A manual glide path was established in first and third groups with #10, 15 and 20 hand K-files. Preparation was performed with reciprocating in-and-out motion, with a 3-4 mm amplitude and slight apical pressure. Initial and final radiographs were taken to analyze the amount of dentin removed in the instrumented canals. The radiographs were superimposed with an image editing software and examined to assess discrepancies at 3-, 6- and 9-mm distances from the apex. The Kruskal-Wallis test was used for statistical analysis. The level of significance was set at 0.05. Results: Preparation in groups without glide paths was swifter than the other groups (P=0.001). However, no difference was observed regarding centering ability. Conclusion: Establishing a glide path increased the total instrumentation time for preparing curved canals with WaveOne and Reciproc instruments. Glide path had no influence on the centering ability of these systems. PMID:26843875

Use of glide-ins in CMS for production and analysis

CERN Document Server

Bradley, D; Hahn, K; Holzman, B; Padhi, S; Pi, H; Spiga, D; Sfiligoi, I; Vaandering, E; Würthwein, F

2010-01-01

With the evolution of various grid federations, the Condor glide-ins represent a key feature in providing a homogeneous pool of resources using late-binding technology. The CMS collaboration uses the glide-in based Workload Management System, glideinWMS, for production (ProdAgent) and distributed analysis (CRAB) of the data. The Condor glide-in daemons traverse to the worker nodes, submitted via Condor-G. Once activated, they preserve the Master-Worker relationships, with the worker first validating the execution environment on the worker node before pulling the jobs sequentially until the expiry of their lifetimes. The combination of late-binding and validation significantly reduces the overall failure rate visible to CMS physicists. We discuss the extensive use of the glideinWMS since the computing challenge, CCRC-08, in order to prepare for the forthcoming LHC data-taking period. The key features essential to the success of large-scale production and analysis on CMS resources across major grid federations,...

A cyclic GMP signalling module that regulates gliding motility in a malaria parasite.

Directory of Open Access Journals (Sweden)

Robert W Moon

2009-09-01

Full Text Available The ookinete is a motile stage in the malaria life cycle which forms in the mosquito blood meal from the zygote. Ookinetes use an acto-myosin motor to glide towards and penetrate the midgut wall to establish infection in the vector. The regulation of gliding motility is poorly understood. Through genetic interaction studies we here describe a signalling module that identifies guanosine 3', 5'-cyclic monophosphate (cGMP as an important second messenger regulating ookinete differentiation and motility. In ookinetes lacking the cyclic nucleotide degrading phosphodiesterase delta (PDEdelta, unregulated signalling through cGMP results in rounding up of the normally banana-shaped cells. This phenotype is suppressed in a double mutant additionally lacking guanylyl cyclase beta (GCbeta, showing that in ookinetes GCbeta is an important source for cGMP, and that PDEdelta is the relevant cGMP degrading enzyme. Inhibition of the cGMP-dependent protein kinase, PKG, blocks gliding, whereas enhanced signalling through cGMP restores normal gliding speed in a mutant lacking calcium dependent protein kinase 3, suggesting at least a partial overlap between calcium and cGMP dependent pathways. These data demonstrate an important function for signalling through cGMP, and most likely PKG, in dynamically regulating ookinete gliding during the transmission of malaria to the mosquito.

Cytogenetic status and oxidative DNA-damage induced by atorvastatin in human peripheral blood lymphocytes: Standard and Fpg-modified comet assay

International Nuclear Information System (INIS)

Gajski, Goran; Garaj-Vrhovac, Vera; Orescanin, Visnja

2008-01-01

To investigate the genotoxic potential of atorvastatin on human lymphocytes in vitro standard comet assay was used in the evaluation of basal DNA damage and to investigate possible oxidative DNA damage produced by reactive oxygen species (ROS) Fpg-modified version of comet assay was also conducted. In addition to these techniques the new criteria for scoring micronucleus test were applied for more complete detection of baseline damage in binuclear lymphocytes exposed to atorvastatin 80 mg/day in different time periods by virtue of measuring the frequency of micronuclei, nucleoplasmic bridges and nuclear buds. All parameters obtained with the standard comet assay and Fpg-modified comet assay were significantly higher in the treated than in control lymphocytes. The Fpg-modified comet assay showed a significantly greater tail length, tail intensity, and tail moment in all treated lymphocytes than did the standard comet assay, which suggests that oxidative stress is likely to be responsible for DNA damage. DNA damage detected by the standard comet assay indicates that some other mechanism is also involved. In addition to the comet assay, a total number of micronuclei, nucleoplasmic bridges and nuclear buds were significantly higher in the exposed than in controlled lymphocytes. Regression analyses showed a positive correlation between the results obtained by the comet (Fpg-modified and standard) and micronucleus assay. Overall, the study demonstrated that atorvastatin in its highest dose is capable of producing damage on the level of DNA molecule and cell

Cytogenetic status and oxidative DNA-damage induced by atorvastatin in human peripheral blood lymphocytes: Standard and Fpg-modified comet assay

Energy Technology Data Exchange (ETDEWEB)

Gajski, Goran [Institute for Medical Research and Occupational Health, Mutagenesis Unit, 10000 Zagreb (Croatia); Garaj-Vrhovac, Vera [Institute for Medical Research and Occupational Health, Mutagenesis Unit, 10000 Zagreb (Croatia); Orescanin, Visnja [Ruder Boskovic Institute, 10000 Zagreb (Croatia)

2008-08-15

To investigate the genotoxic potential of atorvastatin on human lymphocytes in vitro standard comet assay was used in the evaluation of basal DNA damage and to investigate possible oxidative DNA damage produced by reactive oxygen species (ROS) Fpg-modified version of comet assay was also conducted. In addition to these techniques the new criteria for scoring micronucleus test were applied for more complete detection of baseline damage in binuclear lymphocytes exposed to atorvastatin 80 mg/day in different time periods by virtue of measuring the frequency of micronuclei, nucleoplasmic bridges and nuclear buds. All parameters obtained with the standard comet assay and Fpg-modified comet assay were significantly higher in the treated than in control lymphocytes. The Fpg-modified comet assay showed a significantly greater tail length, tail intensity, and tail moment in all treated lymphocytes than did the standard comet assay, which suggests that oxidative stress is likely to be responsible for DNA damage. DNA damage detected by the standard comet assay indicates that some other mechanism is also involved. In addition to the comet assay, a total number of micronuclei, nucleoplasmic bridges and nuclear buds were significantly higher in the exposed than in controlled lymphocytes. Regression analyses showed a positive correlation between the results obtained by the comet (Fpg-modified and standard) and micronucleus assay. Overall, the study demonstrated that atorvastatin in its highest dose is capable of producing damage on the level of DNA molecule and cell.

Kinematics of ram filter feeding and beat-glide swimming in the northern anchovy Engraulis mordax.

Science.gov (United States)

Carey, Nicholas; Goldbogen, Jeremy A

2017-08-01

In the dense aquatic environment, the most adept swimmers are streamlined to reduce drag and increase the efficiency of locomotion. However, because they open their mouth to wide gape angles to deploy their filtering apparatus, ram filter feeders apparently switch between diametrically opposite swimming modes: highly efficient, streamlined 'beat-glide' swimming, and ram filter feeding, which has been hypothesized to be a high-cost feeding mode because of presumed increased drag. Ram filter-feeding forage fish are thought to play an important role in the flux of nutrients and energy in upwelling ecosystems; however, the biomechanics and energetics of this feeding mechanism remain poorly understood. We quantified the kinematics of an iconic forage fish, the northern anchovy, Engraulis mordax , during ram filter feeding and non-feeding, mouth-closed beat-glide swimming. Although many kinematic parameters between the two swimming modes were similar, we found that swimming speeds and tailbeat frequencies were significantly lower during ram feeding. Rather than maintain speed with the school, a speed which closely matches theoretical optimum filter-feeding speeds was consistently observed. Beat-glide swimming was characterized by high variability in all kinematic parameters, but variance in kinematic parameters was much lower during ram filter feeding. Under this mode, body kinematics are substantially modified, and E. mordax swims more slowly and with decreased lateral movement along the entire body, but most noticeably in the anterior. Our results suggest that hydrodynamic effects that come with deployment of the filtering anatomy may limit behavioral options during foraging and result in slower swimming speeds during ram filtration. © 2017. Published by The Company of Biologists Ltd.

Adhesion improvement of glass-fibre-reinforced polyester composites by gliding arc discharge treatment

DEFF Research Database (Denmark)

Kusano, Yukihiro; Sørensen, Bent F.; Løgstrup Andersen, Tom

2013-01-01

A gliding arc is a plasma that can be operated at atmospheric pressure and applied for plasma surface treatment for adhesion improvement. In the present work, glass-fibre-reinforced polyester plates were treated using an atmospheric pressure gliding arc discharge with an air flow to improve...

The collapse of stacking-fault tetrahedra by interaction with gliding dislocations

International Nuclear Information System (INIS)

Matsukawa, Y.; Osetsky, Yu.N.; Stoller, R.E.; Zinkle, S.J.

2005-01-01

The collapse of stacking-fault tetrahedra (SFT) by gliding dislocations was observed in in situ straining experiments in a transmission electron microscope (TEM). A stacking-fault tetrahedron was collapsed by intersection with a gliding perfect dislocation: only the base portion divided by the gliding plane of the dislocation annihilated, while the apex portion remained intact. As a result of analysis on evolution of atom configuration induced by intersection with perfect dislocation in SFT, it was found that an unusual atom configuration inevitably appeared in one of the ledges formed on stacking-fault planes, which is traditionally called I-ledge: the atoms on adjacent (1 1 1) planes were overlapping each other. The overlapping configuration provides a strong repulsive force, being a conceivable driving force to induce a chain reaction of atom displacements that collapses the SFT base portion

Detection of Hypoxia in Human Brain Tumor Xenografts Using a Modified Comet Assay

Directory of Open Access Journals (Sweden)

Jingli Wang

2003-07-01

Full Text Available We used the standard comet assay successfully to generate in vitro dose-response curves under oxic and hypoxic conditions. We then made mixtures of cells that had been irradiated with 3 and 9 Gy of X-rays to simulate two subpopulations in a tumor, but efforts to accurately detect and quantify the subpopulations using the standard comet assay were unsuccessful. Therefore, we investigated a modified comet assay to determine whether it could be used for measuring hypoxia in our model systems. U251 MG cells were grown as subcutaneous tumors in athymic mice; U251 MG and U87 MG cells were grown as intracerebral (i.c. tumors in athymic rats. Animals were injected with RSU 1069, irradiated, and euthanized. Tumors and normal brains were removed, and the cells were analyzed using a modified comet assay. Differences in comet tail moment distributions between tumor and contralateral normal brain, using tail moments at either the 25th or 50th percentile in each distribution, were taken as measures of the degree of tumor hypoxia. For U251 MG tumors, there was a positive relationship between tumor size and the degree of hypoxia, whereas preliminary data from U87 MG i.c. tumors showed less hypoxia and no apparent relationship between tumor size and hypoxia.

Root canal anatomy preservation of WaveOne reciprocating files with or without glide path.

Science.gov (United States)

Berutti, Elio; Paolino, Davide Salvatore; Chiandussi, Giorgio; Alovisi, Mario; Cantatore, Giuseppe; Castellucci, Arnaldo; Pasqualini, Damiano

2012-01-01

This study evaluated the influence of glide path on canal curvature and axis modification after instrumentation with WaveOne Primary reciprocating files. Thirty ISO 15, 0.02 taper Endo Training Blocks were used. In group 1, glide path was created with PathFile 1, 2, and 3 at working length, whereas in group 2, glide path was not performed. In both groups, canals were shaped with WaveOne Primary reciprocating files at working length. Preinstrumentation and postinstrumentation digital images were superimposed and processed with Matlab r2010b software to analyze the curvature radius ratio (CRr) and the relative axis error (rAe), representing canal curvature modification. Data were analyzed with 1-way balanced analyses of variance at 2 levels (P < .05). Glide path was found to be extremely significant for both CRr parameter (F = 9.59; df = 1; P = .004) and rAe parameter (F = 13.55; df = 1; P = .001). Canal modifications seem to be significantly reduced when previous glide path is performed by using the new WaveOne nickel-titanium single-file system. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Dislocation glide velocity in creep of Mg alloys derived from dip tests

Czech Academy of Sciences Publication Activity Database

Eisenlohr, P.; Blum, W.; Milička, Karel

510-511, Sp. Iss. (2009), s. 393-397 ISSN 0921-5093. [Creep 2008. Bayreuth, 04.05.2008-09.05.2008] R&D Projects: GA ČR GA106/06/1354 Institutional research plan: CEZ:AV0Z20410507 Keywords : Dislocation glide velocity * Temperature dependence * Solute drag * Forest cutting * Prismatic glide Subject RIV: JG - Metallurgy Impact factor: 1.901, year: 2009

Water-cooled non-thermal gliding arc for adhesion improvement of glass-fibre-reinforced polyester

DEFF Research Database (Denmark)

Kusano, Yukihiro; Sørensen, Bent F.; Løgstrup Andersen, Tom

2013-01-01

A non-equilibrium quenched plasma is prepared using a gliding-arc discharge generated between diverging electrodes and extended by a gas flow. It can be operated at atmospheric pressure and applied to plasma surface treatment to improve adhesion properties of material surfaces. In this work, glass......-fibre-reinforced polyester plates were treated using an atmospheric pressure gliding-arc discharge with air flow to improve adhesion with a vinylester adhesive. The electrodes were water-cooled so as to operate the gliding arc continually. The treatment improved wettability and increased the density of oxygen...

Modified in vivo comet assay detects the genotoxic potential of 14-hydroxycodeinone, an α,β-unsaturated ketone in oxycodone.

Science.gov (United States)

Pant, Kamala; Roden, Nicholas; Zhang, Charles; Bruce, Shannon; Wood, Craig; Pendino, Kimberly

2015-12-01

14-Hydroxycodeinone (14-HC) is an α,β-unsaturated ketone impurity found in oxycodone drug substance and has a structural alert for genotoxicity. 14-HC was tested in a combined Modified and Standard Comet Assay to determine if the slight decrease in % Tail DNA noted in a previously conducted Standard Comet Assay with 14-HC could be magnified to clarify if the response was due to cross-linking activity. One limitation of the Standard Comet Assay is that DNA cross-links cannot be reliably detected. However, under certain modified testing conditions, DNA cross-links and chemical moieties that elicit such cross-links can be elucidated. One such modification involves the induction of additional breakages of DNA strands by gamma or X-ray irradiation. To determine if 14-HC is a DNA crosslinker in vivo, a Modified Comet Assay was conducted using X-ray irradiation as the modification to visualize crosslinking activity. In this assay, 14-HC was administered orally to mice up to 320 mg/kg/day. Results showed a statistically significant reduction in percent tail DNA in duodenal cells at 320 mg/kg/day, with a nonstatistically significant but dose-related reduction in percent tail DNA also observed at the mid dose of 160 mg/kg/day. Similar decreases were not observed in cells from the liver or stomach, and no increases in percent tail DNA were noted for any tissue in the concomitantly conducted Standard Comet Assay. Taken together, 14-HC was identified as a cross-linking agent in the duodenum in the Modified Comet Assay. © 2015 Wiley Periodicals, Inc.

Irradiation creep by climb-enables glide of dislocations resulting from preferred absorption of point defects

Energy Technology Data Exchange (ETDEWEB)

Mansur, L K [Oak Ridge National Lab., TN (USA)

1979-04-01

A mechanism of irradiation creep arising from the climb-enabled glide of dislocations due to stress-induced preferred absorption of radiation-produced point defects is proposed. This creep component is here termed preferred absorption glide, PAG. PAG-creep operates in addition to the previously studied components of creep from climb by stress-induced preferred absorption, (SI) PA-creep, and the climb-enabled glide due to excess absorption of interstitials on dislocations during swelling, I-creep. A formulation of the various climb and climb-enabled glide processes which includes earlier results is presented. PAG-creep is comparable in magnitude to PA-creep in the parameter range of applications. While the PSA-creep rate and the I-creep rate are linear in stress, the PAG-creep rate is quadratic in stress and thus dominates at high stresses.

Experimental protocols for and studies of the effects of surface passivation and water isotopes on the gliding speed of microtubules propelled by kinesin-1

Science.gov (United States)

Maloney, Roger Andrew

This dissertation explores how the kinesin-1 and microtubule system is affected by surface passivation and water isotopes. Surface passivation was found to affect the gliding speed that microtubules exhibit in the gliding motility assay and the lengths of microtubules supported by the passivation. It was also found that gliding speeds of microtubules are very sensitive to temperature changes. Studies changing the water isotope were a first attempt to investigate if changing the solvent changed the osmotic pressure of the solution kinesin and microtubules were in. No osmotic pressure changes were observed, however, the experiments using different isotopes of water did illuminate the possibility that kinesin may be sensitive to viscosity changes in the solvent. This experiment also suggests further experiments that can be specifically designed to probe osmotic pressure changes. This thesis was also the first thesis ever, to the best of the author's knowledge, to be done in a completely open format. All information and notebook entries that are related to it, as well as the thesis itself, can be found on the website OpenWetWare. The thesis can also be found there including all the different versions that went into its editing. The philosophy and process of making data open and accessible to every one is also discussed.

Long-term vascular contractility assay using genipin-modified muscular thin films

International Nuclear Information System (INIS)

Hald, Eric S; Steucke, Kerianne E; Reeves, Jack A; Win, Zaw; Alford, Patrick W

2014-01-01

Vascular disease is a leading cause of death globally and typically manifests chronically due to long-term maladaptive arterial growth and remodeling. To date, there is no in vitro technique for studying vascular function over relevant disease time courses that both mimics in vivo-like tissue structure and provides a simple readout of tissue stress. We aimed to extend tissue viability in our muscular thin film contractility assay by modifying the polydimethylsiloxane (PDMS) substrate with micropatterned genipin, allowing extracellular matrix turnover without cell loss. To achieve this, we developed a microfluidic delivery system to pattern genipin and extracellular matrix proteins on PDMS prior to cell seeding. Tissues constructed using this method showed improved viability and maintenance of in vivo-like lamellar structure. Functional contractility of tissues fabricated on genipin-modified substrates remained consistent throughout two weeks in culture. These results suggest that muscular thin films with genipin-modified PDMS substrates are a viable method for conducting functional studies of arterial growth and remodeling in vascular diseases. (paper)

The 3D CFD study of gliding swimmer on passive hydrodynamics drag

Directory of Open Access Journals (Sweden)

Vishveshwar Rajendra Mantha

2014-04-01

Full Text Available The aim of this study was to analyze the effect of depth on the hydrodynamic drag coefficient during the passive underwater gliding after the starts and turns. The swimmer hydrodynamics performance was studied by the application of computational fluid dynamics (CFD method. The steady-state CFD simulations were performed by the application of k - omega turbulent model and volume of fluid method to obtain two-phase flow around a three-dimensional swimmer model when gliding near water surface and at different depths from the water surface. The simulations were conducted for four different swimming pool size, each with different depth, i.e., 1.0, 1.5, 2.0 and 3.0 m for three different velocities, i.e., 1.5, 2.0 and 2.5 m/s, with swimmer gliding at different depths with intervals of 0.25 m, each starting from the water surface, respectively. The numerical results of pressure drag and total coefficients at individual average race velocities were obtained. The results showed that the drag coefficient decreased as depth increased, with a trend toward reduced fluctuation after 0.5m depth from the water surface. The selection of the appropriate depth during the gliding phase should be a main concern of swimmers and coaches.

Association of manual or engine-driven glide path preparation with canal centring and apical transportation: a systematic review.

Science.gov (United States)

Hartmann, R C; Peters, O A; de Figueiredo, J A P; Rossi-Fedele, G

2018-04-28

The role and effect of glide path preparation in root canal treatment remain controversial. This systematic review aims to compare apical transportation and canal centring of different glide path preparation techniques, with or without subsequent engine-driven root canal preparation. A database search in PubMed, PubMed Central, Embase, Scopus, EBSCO Dentistry & Oral Sciences Source and Virtual Health Library was conducted, using appropriate key words to identify the effect of glide path preparation (or its absence) on apical transportation and canal centring. An assessment for the risk of bias in included studies was carried out. Amongst 2146 studies, 18 satisfied the inclusion criteria. Nine studies assessed glide path preparation per se, comparing apical transportation and canal centring of rotary systems and/or manual files; eleven further investigations examined the efficacy of the glide path prior to final canal preparation with different engine-driven systems. Risk of bias and other study design features with potential influence on study outcomes and clinical implications were assessed. Based on the available evidence, and within the limitation of the studies included, preparation of a glide path using rotary sequences performs similarly (in most of the component studies) or significantly better than manual preparation when assessing apical transportation or canal centring. When compared to the absence of a glide path, canal shaping following glide path preparation was of similar, or significantly better quality, in regard to apical transportation or canal centring. © 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Characteristics of Atmospheric Pressure Rotating Gliding Arc Plasmas

Science.gov (United States)

Zhang, Hao; Zhu, Fengsen; Tu, Xin; Bo, Zheng; Cen, Kefa; Li, Xiaodong

2016-05-01

In this work, a novel direct current (DC) atmospheric pressure rotating gliding arc (RGA) plasma reactor has been developed for plasma-assisted chemical reactions. The influence of the gas composition and the gas flow rate on the arc dynamic behaviour and the formation of reactive species in the N2 and air gliding arc plasmas has been investigated by means of electrical signals, high speed photography, and optical emission spectroscopic diagnostics. Compared to conventional gliding arc reactors with knife-shaped electrodes which generally require a high flow rate (e.g., 10-20 L/min) to maintain a long arc length and reasonable plasma discharge zone, in this RGA system, a lower gas flow rate (e.g., 2 L/min) can also generate a larger effective plasma reaction zone with a longer arc length for chemical reactions. Two different motion patterns can be clearly observed in the N2 and air RGA plasmas. The time-resolved arc voltage signals show that three different arc dynamic modes, the arc restrike mode, takeover mode, and combined modes, can be clearly identified in the RGA plasmas. The occurrence of different motion and arc dynamic modes is strongly dependent on the composition of the working gas and gas flow rate. supported by National Natural Science Foundation of China (No. 51576174), the Specialized Research Fund for the Doctoral Program of Higher Education of China (No. 20120101110099) and the Fundamental Research Funds for the Central Universities (No. 2015FZA4011)

High throughput analysis of red wine and grape phenolics-adaptation and validation of methyl cellulose precipitable tannin assay and modified Somers color assay to a rapid 96 well plate format.

Science.gov (United States)

Mercurio, Meagan D; Dambergs, Robert G; Herderich, Markus J; Smith, Paul A

2007-06-13

The methyl cellulose precipitable (MCP) tannin assay and a modified version of the Somers and Evans color assay were adapted to high-throughput (HTP) analysis. To improve efficiency of the MCP tannin assay, a miniaturized 1 mL format and a HTP format using 96 well plates were developed. The Somers color assay was modified to allow the standardization of pH and ethanol concentrations of wine samples in a simple one-step dilution with a buffer solution, thus removing inconsistencies between wine matrices prior to analysis and allowing for its adaptation to a HTP format. Validation studies showed that all new formats were efficient, and results were reproducible and analogous to the original formats.

Contact- and Protein Transfer-Dependent Stimulation of Assembly of the Gliding Motility Machinery in Myxococcus xanthus.

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Beata Jakobczak

2015-07-01

Full Text Available Bacteria engage in contact-dependent activities to coordinate cellular activities that aid their survival. Cells of Myxococcus xanthus move over surfaces by means of type IV pili and gliding motility. Upon direct contact, cells physically exchange outer membrane (OM lipoproteins, and this transfer can rescue motility in mutants lacking lipoproteins required for motility. The mechanism of gliding motility and its stimulation by transferred OM lipoproteins remain poorly characterized. We investigated the function of CglC, GltB, GltA and GltC, all of which are required for gliding. We demonstrate that CglC is an OM lipoprotein, GltB and GltA are integral OM β-barrel proteins, and GltC is a soluble periplasmic protein. GltB and GltA are mutually stabilizing, and both are required to stabilize GltC, whereas CglC accumulate independently of GltB, GltA and GltC. Consistently, purified GltB, GltA and GltC proteins interact in all pair-wise combinations. Using active fluorescently-tagged fusion proteins, we demonstrate that GltB, GltA and GltC are integral components of the gliding motility complex. Incorporation of GltB and GltA into this complex depends on CglC and GltC as well as on the cytoplasmic AglZ protein and the inner membrane protein AglQ, both of which are components of the gliding motility complex. Conversely, incorporation of AglZ and AglQ into the gliding motility complex depends on CglC, GltB, GltA and GltC. Remarkably, physical transfer of the OM lipoprotein CglC to a ΔcglC recipient stimulates assembly of the gliding motility complex in the recipient likely by facilitating the OM integration of GltB and GltA. These data provide evidence that the gliding motility complex in M. xanthus includes OM proteins and suggest that this complex extends from the cytoplasm across the cell envelope to the OM. These data add assembly of gliding motility complexes in M. xanthus to the growing list of contact-dependent activities in bacteria.

Adenovirus 2, Bordetella bronchiseptica, and Parainfluenza Molecular Diagnostic Assay Results in Puppies After vaccination with Modified Live Vaccines.

Science.gov (United States)

Ruch-Gallie, R; Moroff, S; Lappin, M R

2016-01-01

Canine adenovirus 2, parainfluenza, and Bordetella bronchiseptica cause respiratory disease in dogs, and each has a modified live intranasal vaccine available. Molecular diagnostic assays to amplify specific nucleic acids are available for each of these agents. If positive molecular diagnostic assay results are common after vaccination, the positive predictive value of the diagnostic assays for disease would be decreased. To determine the impact of administration of commercially available modified live topical adenovirus 2, B. bronchiseptica, and parainfluenza vaccine has on the results of a commercially available PCR panel. Eight puppies from a research breeding facility negative for these pathogens. Blinded prospective pilot study. Puppies were vaccinated with a single dose of modified live topical adenovirus 2, B. bronchiseptica, and parainfluenza and parenteral dose of adenovirus 2, canine distemper virus, and parvovirus. Nasal and pharyngeal swabs were collected on multiple days and submitted for PCR assay. Nucleic acids of all 3 organisms contained in the topical vaccine were detected from both samples multiple times through 28 days after vaccination with higher numbers of positive samples detected between days 3 and 10 after vaccination. Vaccine status should be considered when interpreting respiratory agent PCR results if modified live vaccines have been used. Development of quantitative PCR and wild-type sequencing are necessary to improve positive predictive value of these assays by distinguishing vaccinate from natural infection. Copyright © 2015 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Comparison of apical extrusion of intracanal bacteria by various glide-path establishing systems: an in vitro study

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Alberto Dagna

2017-11-01

Full Text Available Objectives This study compared the amount of apically extruded bacteria during the glide-path preparation by using multi-file and single-file glide-path establishing nickel-titanium (NiTi rotary systems. Materials and Methods Sixty mandibular first molar teeth were used to prepare the test apparatus. They were decoronated, blocked into glass vials, sterilized in ethylene oxide gas, infected with a pure culture of Enterococcus faecalis, randomly assigned to 5 experimental groups, and then prepared using manual stainless-steel files (group KF and glide-path establishing NiTi rotary files (group PF with PathFiles, group GF with G-Files, group PG with ProGlider, and group OG with One G. At the end of canal preparation, 0.01 mL NaCl solution was taken from the experimental vials. The suspension was plated on brain heart infusion agar and colonies of bacteria were counted, and the results were given as number of colony-forming units (CFU. Results The manual instrumentation technique tested in group KF extruded the highest number of bacteria compared to the other 4 groups (p < 0.05. The 4 groups using rotary glide-path establishing instruments extruded similar amounts of bacteria. Conclusions All glide-path establishment instrument systems tested caused a measurable apical extrusion of bacteria. The manual glide-path preparation showed the highest number of bacteria extruded compared to the other NiTi glide-path establishing instruments.

Nitrogen Fixation by Gliding Arc Plasma: Better Insight by Chemical Kinetics Modelling.

Science.gov (United States)

Wang, Weizong; Patil, Bhaskar; Heijkers, Stjin; Hessel, Volker; Bogaerts, Annemie

2017-05-22

The conversion of atmospheric nitrogen into valuable compounds, that is, so-called nitrogen fixation, is gaining increased interest, owing to the essential role in the nitrogen cycle of the biosphere. Plasma technology, and more specifically gliding arc plasma, has great potential in this area, but little is known about the underlying mechanisms. Therefore, we developed a detailed chemical kinetics model for a pulsed-power gliding-arc reactor operating at atmospheric pressure for nitrogen oxide synthesis. Experiments are performed to validate the model and reasonable agreement is reached between the calculated and measured NO and NO 2 yields and the corresponding energy efficiency for NO x formation for different N 2 /O 2 ratios, indicating that the model can provide a realistic picture of the plasma chemistry. Therefore, we can use the model to investigate the reaction pathways for the formation and loss of NO x . The results indicate that vibrational excitation of N 2 in the gliding arc contributes significantly to activating the N 2 molecules, and leads to an energy efficient way of NO x production, compared to the thermal process. Based on the underlying chemistry, the model allows us to propose solutions on how to further improve the NO x formation by gliding arc technology. Although the energy efficiency of the gliding-arc-based nitrogen fixation process at the present stage is not comparable to the world-scale Haber-Bosch process, we believe our study helps us to come up with more realistic scenarios of entering a cutting-edge innovation in new business cases for the decentralised production of fertilisers for agriculture, in which low-temperature plasma technology might play an important role. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Influence of a glide path on the dentinal crack formation of ProTaper Next system

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Sevinç Aktemur Türker

2015-11-01

Full Text Available Objectives The aim was to evaluate dentinal crack formation after root canal preparation with ProTaper Next system (PTN with and without a glide path. Materials and Methods Forty-five mesial roots of mandibular first molars were selected. Fifteen teeth were left unprepared and served as controls. The experimental groups consist of mesiobuccal and mesiolingual root canals of remaining 30 teeth, which were divided into 2 groups (n = 15: Group PG/PTN, glide path was created with ProGlider (PG and then canals were shaped with PTN system; Group PTN, glide path was not prepared and canals were shaped with PTN system only. All roots were sectioned perpendicular to the long axis at 1, 2, 3, 4, 6, and 8 mm from the apex, and the sections were observed under a stereomicroscope. The presence/absence of cracks was recorded. Data were analyzed with chi-square tests with Yates correction. Results There were no significant differences in crack formation between the PTN with and without glide path preparation. The incidence of cracks observed in PG/PTN and PTN groups was 17.8% and 28.9%, respectively. Conclusions The creation of a glide path with ProGlider before ProTaper Next rotary system did not influence dentinal crack formation in root canals.

Is Snow Gliding a Major Soil Erosion Agent in Steep Alpine Areas?

International Nuclear Information System (INIS)

Meusburger, K.; Walter, A.; Alewell, C.; Leitinger, G.; Mabit, L.; Mueller, M.H.

2015-01-01

Snow cover is a key hydrological characteristic of mountain areas. Nevertheless, a majority of studies focused on quantifying rates of soil erosion and sediment transport in steep mountain areas has largely neglected the role of snow cover on soil erosion rates (Stanchi et al., 2014). Soil erosion studies have focused almost exclusively on the snow-free periods even though it is well known that wet avalanches can yield enormous erosive forces (Freppaz et al., 2010; Korup and Rixen, 2014). This raises the question whether annual snow cover and particularly the slow movement of snow packages over the soil surface, termed ‘‘snow gliding’’, contribute significantly to the total soil loss in these areas. Three different approaches to estimate soil erosion rates were used to address this question. These include (1) the anthropogenic soil tracer 137 Cs, (2) the Revised Universal Soil Loss Equation (RUSLE), and (3) direct sediment yield measurements of snow glide deposits. The fallout radionuclide 137 Cs integrates total soil loss due to all erosion agents involved, the RUSLE model is suitable to estimate soil loss by water erosion and the sediment yield measurements yield represents a direct estimate of soil removal by snow gliding. Moreover, cumulative snow glide distance was measured for 14 sites and modelled for the surrounding area with the Spatial Snow Glide Model (Leitinger et al., 2008)

Gliding lizards use the position of the sun to enhance social display.

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Klomp, Danielle A; Stuart-Fox, Devi; Das, Indraneil; Ord, Terry J

2017-02-01

Effective communication requires animal signals to be readily detected by receivers in the environments in which they are typically given. Certain light conditions enhance the visibility of colour signals and these conditions can vary depending on the orientation of the sun and the position of the signaller. We tested whether Draco sumatranus gliding lizards modified their position relative to the sun to enhance the conspicuousness of their throat-fan (dewlap) during social display to conspecifics. The dewlap was translucent, and we found that lizards were significantly more likely to orient themselves perpendicular to the sun when displaying. This increases the dewlap's radiance, and likely, its conspicuousness, by increasing the amount of light transmitted through the ornament. This is a rare example of a behavioural adaptation for enhancing the visibility of an ornament to distant receivers. © 2017 The Author(s).

Influence of GlideScope assisted endotracheal intubation on intraocular pressure in ophthalmic patients

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Nauman Ahmad

2015-01-01

Full Text Available Background: Traditional Macintoch laryngoscopy is known to cause a rise in intraocular pressure (IOP, tachycardia and hypertension. These changes are not desirable in patients with glaucoma and open globe injury. GlideScope is a video laryngoscope that functions independent of the line of sight, reduces upward lifting forces for glottic exposure and requires less cervical neck movement for intubation, making it less stimulating than Macintosh laryngoscopy. Aim: The aim was to assess the variations in IOP and hemodynamic changes after GlideScope assisted intubation. Materials and Methods: After approval of the local Institutional Research and Ethical Board and informed patient consent, 50 adult American Society of Anesthesiologist I and II patients with normal IOP were enrolled in a prospective, randomized study for ophthalmic surgery requiring tracheal intubation. In all patients, trachea was intubated using either GlideScope or Macintoch laryngoscope. IOP of nonoperated eye, heart rate and blood pressure were measured as baseline, 1 min after induction, 1 min and 5 min after tracheal intubation. Results: IOP was not significantly different between groups before and after anesthetic induction and 5 min after tracheal intubation (P = 0.217, 0.726, and 0.110 respectively. The only significant difference in IOP was at 1 min after intubation (P = 0.041. No significant difference noted between groups in mean arterial pressure (P = 0.899, 0.62, 0.47, 0.82 respectively and heart rate (P = 0.21, 0.72, 0.07, 0.29, respectively at all measurements. Conclusion: GlideScope assisted tracheal intubation shown lesser rise in IOP at 1 min after intubation in comparison to Macintoch laryngoscope, suggesting that GlideScope may be preferable to Macintosh laryngoscope.

Gliding Swifts Attain Laminar Flow over Rough Wings

NARCIS (Netherlands)

Lentink, D.; Kat, de R.

2014-01-01

Swifts are among the most aerodynamically refined gliding birds. However, the overlapping vanes and protruding shafts of their primary feathers make swift wings remarkably rough for their size. Wing roughness height is 1–2% of chord length on the upper surface—10,000 times rougher than sailplane

Dislocation glide in Ni-Al solid solutions from the atomic scale up: a molecular dynamics study

International Nuclear Information System (INIS)

Rodary, E.

2003-01-01

The glide of an edge dislocation in solid solutions is studied by molecular dynamics, at fixed temperature and imposed external stress. We have optimized an EAM potential for Ni(1 a 8% A1): it well reproduces the lattice expansion, local atomic order, stacking fault energy as a function of composition, as well as the elastic properties of the γ' phase with L1 2 structure. On increasing the stress, the dislocation is first immobile, then glides with a velocity proportional to the stress and the velocity saturates on reaching the transverse sound velocity. However, only beyond a static threshold stress, σ s , does the dislocation glide a distance large enough to allow macroscopic shear; the linear part of the velocity-stress curve extrapolates to zero at a dynamical threshold stress, σ d , The friction coefficient, and the threshold stresses (σ s and σ d ), increase with the A1 concentration and decrease with temperature (300 and 500 K). Close to the critical shear stress, σ s , the dislocation glide is analysed with a 'stop and go' model. The latter yields the flight velocity between obstacles, the mean obstacle density and the distribution of the waiting time on each obstacle as a function of stress, composition and temperature. The obstacle to the glide is proposed to be the strong repulsion between Al atoms brought into nearest neighbour position by the glide process, and not the dislocation-solute interaction. The microscopic parameters so defined are introduced into a micro-mechanical model, which well reproduces the known behaviour of nickel base solid solutions. (author)

Comparison of patellar distraction with patellar glides in female patients with patellofemoral pain syndrom

International Nuclear Information System (INIS)

Syed, S.; Chaudhary, M.A.; Noor, R.; Bashir, M.S.; Manzoor, B.

2017-01-01

To analyse effectiveness of patellar glides and patellar distraction in the patients with patellofemoral pain syndrome (PFPS). Methodology: This longitudinal interventional comparative study was conducted at Physiotherapy Department, Mayo Hospital Lahore, Pakistan from September 2015 to March 2016. A total of 70 patients were divided into 2 groups randomly; group A received hot pack, quadriceps strengthening exercises and patellar distraction whereas group B received hot pack, quadriceps strengthening exercises and patellar glides. Age of the female patients was 18-40 years. Visual Analogue Scale (VAS) and Knee Injury and osteoarthritis outcome score (KOOS) questionnaire were used to compare the effectiveness of both treatments. The data were analysed using SPSS v. 21.0. Results: There was reduction in pain at VAS and KOOS showed improvement in function as well as the range of motion also increased in both groups. Both treatment techniques were effective in reducing pain in PFPS (P <0.005). Pre-treatment KOOS score in patellar glides group was 34.77+10.84 and post-treatment KOOS score was 62.155+15.75 and for patellar distraction group pre-treatment KOOS score was 35.42+10.07 that increased to 55.77+14.66 after treatment which showed that patellar glides had better effect on PFPS. Conclusion: Both treatments were effective in managing PFPS in terms of decreasing pain and increasing ROM as there was no significant difference between two techniques, however patellar glides were superior as compared to patellar distraction in decreasing pain and increasing ROM. (author)

Aerodynamic characteristics of flying fish in gliding flight.

Science.gov (United States)

Park, Hyungmin; Choi, Haecheon

2010-10-01

The flying fish (family Exocoetidae) is an exceptional marine flying vertebrate, utilizing the advantages of moving in two different media, i.e. swimming in water and flying in air. Despite some physical limitations by moving in both water and air, the flying fish has evolved to have good aerodynamic designs (such as the hypertrophied fins and cylindrical body with a ventrally flattened surface) for proficient gliding flight. Hence, the morphological and behavioral adaptations of flying fish to aerial locomotion have attracted great interest from various fields including biology and aerodynamics. Several aspects of the flight of flying fish have been determined or conjectured from previous field observations and measurements of morphometric parameters. However, the detailed measurement of wing performance associated with its morphometry for identifying the characteristics of flight in flying fish has not been performed yet. Therefore, in the present study, we directly measure the aerodynamic forces and moment on darkedged-wing flying fish (Cypselurus hiraii) models and correlated them with morphological characteristics of wing (fin). The model configurations considered are: (1) both the pectoral and pelvic fins spread out, (2) only the pectoral fins spread with the pelvic fins folded, and (3) both fins folded. The role of the pelvic fins was found to increase the lift force and lift-to-drag ratio, which is confirmed by the jet-like flow structure existing between the pectoral and pelvic fins. With both the pectoral and pelvic fins spread, the longitudinal static stability is also more enhanced than that with the pelvic fins folded. For cases 1 and 2, the lift-to-drag ratio was maximum at attack angles of around 0 deg, where the attack angle is the angle between the longitudinal body axis and the flying direction. The lift coefficient is largest at attack angles around 30∼35 deg, at which the flying fish is observed to emerge from the sea surface. From glide polar

A modified Plasmodium falciparum growth inhibition assay (GIA) to assess activity of plasma from malaria endemic areas.

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Mlambo, Godfree; Kumar, Nirbhay

2007-02-01

Plasma samples from patients undergoing treatment in malaria endemic countries often contain anti-malaria drugs, that may overstate effects of specific antibodies in growth inhibition assays (GIA). We describe a modified assay that uses drug resistant P. falciparum parasites (W2) that circumvents the requirement for dialyzing samples that may likely contain drugs such as chloroquine and sulfadoxine/pyrimethamine (SP).

A rolling-gliding wear simulator for the investigation of tribological material pairings for application in total knee arthroplasty

OpenAIRE

Richter, Berna I; Ostermeier, Sven; Turger, Anke; Denkena, Berend; Hurschler, Christof

2010-01-01

Abstract Background Material wear testing is an important technique in the development and evaluation of materials for use in implant for total knee arthroplasty. Since a knee joint induces a complex rolling-gliding movement, standardised material wear testing devices such as Pin-on-Disc or Ring-on-Disc testers are suitable to only a limited extent because they generate pure gliding motion only. Methods A rolling-gliding wear simulator was thus designed, constructed and implemented, which sim...

Circular random motion in diatom gliding under isotropic conditions

International Nuclear Information System (INIS)

Gutiérrez-Medina, Braulio; Maldonado, Ana Iris Peña; Guerra, Andrés Jiménez; Rubio, Yadiralia Covarrubias; Meza, Jessica Viridiana García

2014-01-01

How cells migrate has been investigated primarily for the case of trajectories composed by joined straight segments. In contrast, little is known when cellular motion follows intrinsically curved paths. Here, we use time-lapse optical microscopy and automated trajectory tracking to investigate how individual cells of the diatom Nitzschia communis glide across surfaces under isotropic environmental conditions. We find a distinct kind of random motion, where trajectories are formed by circular arcs traveled at constant speed, alternated with random stoppages, direction reversals and changes in the orientation of the arcs. Analysis of experimental and computer-simulated trajectories show that the circular random motion of diatom gliding is not optimized for long-distance travel but rather for recurrent coverage of limited surface area. These results suggest that one main biological role for this type of diatom motility is to efficiently build the foundation of algal biofilms. (paper)

High Order Large Eddy Simulation (LES) of Gliding Snake Aerodynamics: Effect of 3D Flow on Gliding Performance

Science.gov (United States)

Delorme, Yann; Hassan, Syed Harris; Socha, Jake; Vlachos, Pavlos; Frankel, Steven

2014-11-01

Chrysopelea paradisi are snakes that are able to glide over long distances by morphing the cross section of their bodies from circular to a triangular airfoil, and undulating through the air. Snake glide is characterized by relatively low Reynolds number and high angle of attack as well as three dimensional and unsteady flow. Here we study the 3D dynamics of the flow using an in-house high-order large eddy simulation code. The code features a novel multi block immersed boundary method to accurately and efficiently represent the complex snake geometry. We investigate the steady state 3-dimensionality of the flow, especially the wake flow induced by the presence of the snake's body, as well as the vortex-body interaction thought to be responsible for part of the lift enhancement. Numerical predictions of global lift and drag will be compared to experimental measurements, as well as the lift distribution along the body of the snake due to cross sectional variations. Comparisons with previously published 2D results are made to highlight the importance of 3-dimensional effects. Additional efforts are made to quantify properties of the vortex shedding and Dynamic Mode Decomposition (DMD) is used to analyse the main modes responsible for the lift and drag forces.

A systematic review on the safety of Prostar XL versus ProGlide after TAVR and EVAR

Energy Technology Data Exchange (ETDEWEB)

Maniotis, Christos [2nd Cardiology Department, Hellenic Red Cross Hospital of Athens, Athens (Greece); Andreou, Constantinos; Karalis, Ioannis [Interventional Cardiology Department, University Hospital of Leiden, Leiden (Netherlands); Koutouzi, Giasemi [Interventional Radiology Department, Sahlgrenska University Hospital, Gothenburg (Sweden); Agelaki, Maria [2nd Cardiology Department, Hellenic Red Cross Hospital of Athens, Athens (Greece); Koutouzis, Michael, E-mail: koutouzismike@yahoo.gr [2nd Cardiology Department, Hellenic Red Cross Hospital of Athens, Athens (Greece)

2017-03-15

Background: Endovascular aortic aneurysm repair (EVAR) and transfemoral transcatheter aortic valve replacement (TAVR) are widely spreading minimally invasive procedures performed mainly through the femoral artery. Prostar XL and ProGlide vascular closure devices are used in clinical practice for the hemostasis in these procedures and they have been shown to be safe and effective. Purpose: The aim of our systematic review is to compare the safety of these two devices for percutaneous closure of large arteriotomies in patients undergoing TAVR and EVAR. Methods: We searched PubMed, EMBASE, Google Scholar and the Cochrane Central Register of Controlled Trials for all randomized and observational published studies that compared Prostar XL vs. ProGlide. Relative risk was calculated by random-effects model. Review Manager 5.1 was used for statistical analysis. Results: A total number of 2909 patients were included in our analysis. The rate of overall vascular complications did not differ between Prostar XL and ProGlide {RR 1.35 (0.80–2.29), p = 0.27}. In contrary, the risk ratio of all bleeding complications with Prostar XL compared to ProGlide was 1.82 (1.47–2.24, p < 0.001) and for major and life-threatening bleeding complications was 2.48 (1.65–3.73, p < 0001, suggesting a lower bleeding risk with ProGlide). No statistical difference was found between groups for end-stage acute kidney injury (AKI), with a risk ratio of 2.14 (0.81–5.66), p = 0.05. Finally, there were no differences in in-hospital and 30-days mortality rate between the two groups (1.41, 0.56–3.54, p = 0.46 and 1.43, 0.55–3.73, p = 0.47, respectively). Conclusions: Prostar XL is associated with greater risk of any bleeding as well as life threatening bleeding compared to the ProGlide device. However, no significant differences were observed in the rate of overall vascular complications, end stage AKI and in-hospital and 30-days mortality. - Highlights: • We present a systematic review

A systematic review on the safety of Prostar XL versus ProGlide after TAVR and EVAR

International Nuclear Information System (INIS)

Maniotis, Christos; Andreou, Constantinos; Karalis, Ioannis; Koutouzi, Giasemi; Agelaki, Maria; Koutouzis, Michael

2017-01-01

Background: Endovascular aortic aneurysm repair (EVAR) and transfemoral transcatheter aortic valve replacement (TAVR) are widely spreading minimally invasive procedures performed mainly through the femoral artery. Prostar XL and ProGlide vascular closure devices are used in clinical practice for the hemostasis in these procedures and they have been shown to be safe and effective. Purpose: The aim of our systematic review is to compare the safety of these two devices for percutaneous closure of large arteriotomies in patients undergoing TAVR and EVAR. Methods: We searched PubMed, EMBASE, Google Scholar and the Cochrane Central Register of Controlled Trials for all randomized and observational published studies that compared Prostar XL vs. ProGlide. Relative risk was calculated by random-effects model. Review Manager 5.1 was used for statistical analysis. Results: A total number of 2909 patients were included in our analysis. The rate of overall vascular complications did not differ between Prostar XL and ProGlide {RR 1.35 (0.80–2.29), p = 0.27}. In contrary, the risk ratio of all bleeding complications with Prostar XL compared to ProGlide was 1.82 (1.47–2.24, p < 0.001) and for major and life-threatening bleeding complications was 2.48 (1.65–3.73, p < 0001, suggesting a lower bleeding risk with ProGlide). No statistical difference was found between groups for end-stage acute kidney injury (AKI), with a risk ratio of 2.14 (0.81–5.66), p = 0.05. Finally, there were no differences in in-hospital and 30-days mortality rate between the two groups (1.41, 0.56–3.54, p = 0.46 and 1.43, 0.55–3.73, p = 0.47, respectively). Conclusions: Prostar XL is associated with greater risk of any bleeding as well as life threatening bleeding compared to the ProGlide device. However, no significant differences were observed in the rate of overall vascular complications, end stage AKI and in-hospital and 30-days mortality. - Highlights: • We present a systematic review

Assessment of the Implementation of GNSS into Gliding

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Tomáš Kubáč

2017-11-01

Full Text Available Global navigation satellite systems are increasingly part of our lives and many industries including aviation. Glider flying is no exception in this trend. Global navigation satellite systems were part of gliding since the early 1990s. First as official recording devices for simple evidence of sporting performances, then as navigation systems, anti-collision systems and emergency location transmitters. Development of recording application was initiated and supported by International Gliding Commission of World Air Sports Federation in way of certifications for flight recorders. The use of navigation and other modern instruments in gliders has brought many benefits but also risks. However, the advantages outweigh the disadvantages and these systems are now integral part of gliding. With this wide usage of global navigation satellite systems devices, there is great many possibilities how and in which way one can use these systems. Pilots must orient themselves in varied selection of products, which they can use to choose one solution, that fits him. Therefore, to find out how and if pilots use these devices, we created questionnaire survey among 143 Czech glider pilots. We found out, that 84% of them are using global navigation satellite systems devices for official record of flight and for navigation as well. More than half of pilots is using free, not built-in devices. Most common devices are mobile phones up to 5 inches of screen diagonal in combination with approved flight recorder without display. If pilots use mobile device for navigation, 52% of them is using one with Windows Mobile operating system, 33% use Android. Navigational software on these mobile devices is then almost tied between SeeYou Mobile, XCSoar and LK8000. Knowledge about usage preference of global navigation systems devices should help pilots with selection and overall orientation in subject.

Using physical models to study the gliding performance of extinct animals.

Science.gov (United States)

Koehl, M A R; Evangelista, Dennis; Yang, Karen

2011-12-01

Aerodynamic studies using physical models of fossil organisms can provide quantitative information about how performance of defined activities, such as gliding, depends on specific morphological features. Such analyses allow us to rule out hypotheses about the function of extinct organisms that are not physically plausible and to determine if and how specific morphological features and postures affect performance. The purpose of this article is to provide a practical guide for the design of dynamically scaled physical models to study the gliding of extinct animals using examples from our research on the theropod dinosaur, †Microraptor gui, which had flight feathers on its hind limbs as well as on its forelimbs. Analysis of the aerodynamics of †M. gui can shed light on the design of gliders with large surfaces posterior to the center of mass and provide functional information to evolutionary biologists trying to unravel the origins of flight in the dinosaurian ancestors and sister groups to birds. Measurements of lift, drag, side force, and moments in pitch, roll, and yaw on models in a wind tunnel can be used to calculate indices of gliding and parachuting performance, aerodynamic static stability, and control effectiveness in maneuvering. These indices permit the aerodynamic performance of bodies of different shape, size, stiffness, texture, and posture to be compared and thus can provide insights about the design of gliders, both biological and man-made. Our measurements of maximum lift-to-drag ratios of 2.5-3.1 for physical models of †M. gui suggest that its gliding performance was similar to that of flying squirrels and that the various leg postures that might have been used by †M. gui make little difference to that aspect of aerodynamic performance. We found that body orientation relative to the movement of air past the animal determines whether it is difficult or easy to maneuver.

The Use of a Reciprocating Handpiece to Create a Glide Path in Curved Canals: Comparison with Manual Glide Path Preparation

Science.gov (United States)

2016-06-01

instruments into the root canal system, manufacturers recommend creating a glide path to reduce the risk of instrument fracture due to taper lock . This...Results The two groups had almost identical mean and standard...The groups had identical median values of 85 seconds, and there was no significant difference between the groups (Mann-Whitney U test; p=0.15; two

Effect of condensation temperature glide on the performance of organic Rankine cycles with zeotropic mixture working fluids

International Nuclear Information System (INIS)

Liu, Qiang; Duan, Yuanyuan; Yang, Zhen

2014-01-01

Highlights: • A condensation pressure determination method for ORC with zeotropic mixture is given. • The effects of condensation temperature glide on the ORC performance are analyzed. • Mixture mole fractions for the maximum power output of a geothermal ORC are identified. • The biomass ORC performance with part of the latent heat transferred in the IHE is analyzed. - Abstract: The organic Rankine cycle (ORC) has been widely used to convert low-grade ( 2 M) selected as working fluids for the cogenerative ORC driven by the biomass energy. Two optimal working fluid mole fractions maximize the cycle efficiency, exergy efficiency and net power output for cooling water temperature increases less than the maximum condensation temperature glide, while the highest net power output appears at the higher mole fraction of the more volatile component for the geothermal ORC when the condensation temperature glide of the working fluid mixture matches the cooling water temperature increase. Higher condensation temperature glides result in large thermal loss to the heat sink and exergy destruction in the condenser. There is only one optimal working fluid mole fraction that maximizes the thermal efficiency, exergy efficiency and net power output when the cooling water temperature increase is greater than the condensation temperature glide

Ancient phylogenetic divergence of the enigmatic African rodent Zenkerella and the origin of anomalurid gliding

Directory of Open Access Journals (Sweden)

Steven Heritage

2016-08-01

Full Text Available The “scaly-tailed squirrels” of the rodent family Anomaluridae have a long evolutionary history in Africa, and are now represented by two gliding genera (Anomalurus and Idiurus and a rare and obscure genus (Zenkerella that has never been observed alive by mammalogists. Zenkerella shows no anatomical adaptations for gliding, but has traditionally been grouped with the glider Idiurus on the basis of craniodental similarities, implying that either the Zenkerella lineage lost its gliding adaptations, or that Anomalurus and Idiurus evolved theirs independently. Here we present the first nuclear and mitochondrial DNA sequences of Zenkerella, based on recently recovered whole-body specimens from Bioko Island (Equatorial Guinea, which show unambiguously that Zenkerella is the sister taxon of Anomalurus and Idiurus. These data indicate that gliding likely evolved only once within Anomaluridae, and that there were no subsequent evolutionary reversals. We combine this new molecular evidence with morphological data from living and extinct anomaluromorph rodents and estimate that the lineage leading to Zenkerella has been evolving independently in Africa since the early Eocene, approximately 49 million years ago. Recently discovered fossils further attest to the antiquity of the lineage leading to Zenkerella, which can now be recognized as a classic example of a “living fossil,” about which we know remarkably little. The osteological markers of gliding are estimated to have evolved along the stem lineage of the Anomalurus–Idiurus clade by the early Oligocene, potentially indicating that this adaptation evolved in response to climatic perturbations at the Eocene–Oligocene boundary (∼34 million years ago.

Dynamics, OH distributions and UV emission of a gliding arc at various flow-rates investigated by optical measurements

International Nuclear Information System (INIS)

Zhu, Jiajian; Sun, Zhiwei; Li, Zhongshan; Ehn, Andreas; Aldén, Marcus; Salewski, Mirko; Leipold, Frank; Kusano, Yukihiro

2014-01-01

We demonstrate a plasma discharge which is generated between two diverging electrodes and extended into a gliding arc in non-equilibrium condition by an air flow at atmospheric pressure. Effects of the air flow rates on the dynamics, ground-state OH distributions and spectral characterization of UV emission of the gliding arc were investigated by optical methods. High-speed photography was utilized to reveal flow-rate dependent dynamics such as ignitions, propagation, short-cutting events, extinctions and conversions of the discharge from glowtype to spark-type. Short-cutting events and ignitions occur more frequently at higher flow rates. The anchor points of the gliding arc are mostly steady at the top of the electrodes at lower flow rates whereas at higher flow rates they glide up along the electrodes most of the time. The afterglow of fully developed gliding arcs is observed to decay over hundreds of microseconds after being electronically short-cut by a newly ignited arc. The extinction time decreases with the increase of the flow rate. The frequency of the conversion of a discharge from glow-type to spark-type increases with the flow rate. Additionally, spatial distributions of ground-state OH were investigated using planar laser-induced fluorescence. The results show that the shape, height, intensity and thickness of ground-state OH distribution vary significantly with air flow rates. Finally, UV emission of the gliding arc is measured using optical emission spectroscopy and it is found that the emission intensity of NO γ (A-X), OH (A-X) and N 2 (C-B) increase with the flow rates showing more characteristics of spark-type arcs. The observed phenomena indicate the significance of the interaction between local turbulence and the gliding arc. (paper)

Dynamics, OH distributions and UV emission of a gliding arc at various flow-rates investigated by optical measurements

Science.gov (United States)

Zhu, Jiajian; Sun, Zhiwei; Li, Zhongshan; Ehn, Andreas; Aldén, Marcus; Salewski, Mirko; Leipold, Frank; Kusano, Yukihiro

2014-07-01

We demonstrate a plasma discharge which is generated between two diverging electrodes and extended into a gliding arc in non-equilibrium condition by an air flow at atmospheric pressure. Effects of the air flow rates on the dynamics, ground-state OH distributions and spectral characterization of UV emission of the gliding arc were investigated by optical methods. High-speed photography was utilized to reveal flow-rate dependent dynamics such as ignitions, propagation, short-cutting events, extinctions and conversions of the discharge from glowtype to spark-type. Short-cutting events and ignitions occur more frequently at higher flow rates. The anchor points of the gliding arc are mostly steady at the top of the electrodes at lower flow rates whereas at higher flow rates they glide up along the electrodes most of the time. The afterglow of fully developed gliding arcs is observed to decay over hundreds of microseconds after being electronically short-cut by a newly ignited arc. The extinction time decreases with the increase of the flow rate. The frequency of the conversion of a discharge from glow-type to spark-type increases with the flow rate. Additionally, spatial distributions of ground-state OH were investigated using planar laser-induced fluorescence. The results show that the shape, height, intensity and thickness of ground-state OH distribution vary significantly with air flow rates. Finally, UV emission of the gliding arc is measured using optical emission spectroscopy and it is found that the emission intensity of NO γ (A-X), OH (A-X) and N2 (C-B) increase with the flow rates showing more characteristics of spark-type arcs. The observed phenomena indicate the significance of the interaction between local turbulence and the gliding arc.

Postoperative pain after manual and mechanical glide path: a randomized clinical trial.

Science.gov (United States)

Pasqualini, Damiano; Mollo, Livio; Scotti, Nicola; Cantatore, Giuseppe; Castellucci, Arnaldo; Migliaretti, Giuseppe; Berutti, Elio

2012-01-01

This prospective randomized clinical trial evaluated the incidence of postoperative pain after glide path performed with PathFile (PF) (Dentsply Maillefer, Ballaigues, Switzerland) versus stainless-steel K-file (KF). In 149 subjects, the mechanical glide path was performed with nickel-titanium (NiTi) rotary PF; in 146 subjects, the manual glide path was performed with stainless-steel KFs. Postoperative pain, analgesics consumption, and the number of days to complete pain resolution were evaluated in the following 7 days. An analysis of variance model for repeated measures was used to compare the variation of pain-scale values (P < .05). The Student's t test for continuous variables normally distributed, the nonparametric Mann-Whitney U test for the nonnormally distributed variables, and the chi-square test for dichotomous variables were used (P < .05). Despite homogeneous baseline conditions at diagnosis, tooth type, pain prevalence, and scores, the postoperative pain prevalence curves in PF group evidenced a more favorable trend in terms of time to pain resolution compared with the KF group (P = .004). The difference was also evident in the model adjusted for analgesics consumption in both groups (P = .012). The mean analgesics intake per subject was significantly higher in the KF group (3.7 ± 2.2) compared with the PF group (2 ± 1.7) (P < .001). Mean pain stop values were also significantly higher in the KF group (2.7) compared with the PF group (1.7) (P = .001). The glide path with NiTi Rotary PF leads to less postoperative pain and faster symptom resolution. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Marked colour divergence in the gliding membranes of a tropical lizard mirrors population differences in the colour of falling leaves.

Science.gov (United States)

Klomp, D A; Stuart-Fox, D; Das, I; Ord, T J

2014-12-01

Populations of the Bornean gliding lizard, Draco cornutus, differ markedly in the colour of their gliding membranes. They also differ in local vegetation type (mangrove forest versus lowland rainforest) and consequently, the colour of falling leaves (red and brown/black in mangrove versus green, brown and black in rainforest). We show that the gliding membranes of these lizards closely match the colours of freshly fallen leaves in the local habitat as they appear to the visual system of birds (their probable predators). Furthermore, gliding membranes more closely resembled colours of local fallen leaves than standing foliage or fallen leaves in the other population's habitat. This suggests that the two populations have diverged in gliding membrane coloration to match the colours of their local falling leaves, and that mimicking falling leaves is an adaptation that functions to reduce predation by birds. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Influence of a glide path on apical extrusion of debris during canal preparation using single-file systems in curved canals.

Science.gov (United States)

Topçuoğlu, H S; Düzgün, S; Akpek, F; Topçuoğlu, G; Aktı, A

2016-06-01

To evaluate the effect of a glide path on the amount of apically extruded debris during canal preparation using single-file systems in curved canals. Ninety extracted mandibular molar teeth were randomly assigned to six groups (n = 15 for each group) for canal instrumentation. Endodontic access cavities were prepared in each tooth. In three of the six groups, a glide path was not created whereas a glide path was created using PathFile instruments on the mesial canals of all teeth in the remaining three groups. The mesial canals of the teeth were then instrumented with the following single-file instrument systems: WaveOne, Reciproc and OneShape. Debris extruded apically during instrumentation was collected into pre-weighed Eppendorf tubes. The tubes were then stored in an incubator at 70 °C for 5 days. The weight of the dry extruded debris was established by subtracting the pre-instrumentation and post-instrumentation weight of the Eppendorf tubes for each group. The data obtained were analysed using one-way analysis of variance (anova) and Tukey's post hoc tests. The OneShape file was associated with less debris extrusion than the Reciproc and WaveOne files when canal instrumentation was performed without a glide path (P files (P > 0.05). There was no significant difference amongst the OneShape, Reciproc and WaveOne files when a glide path was created before canal preparation in curved root canals (P > 0.05). All systems extruded significantly less debris in groups with a glide path than in groups without a glide path (P < 0.05). All instruments were associated with apical extrusion of debris. Creating a glide path prior to canal instrumentation reduced the amount of apically extruded debris in curved canals. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

A theoretical analysis of pitch stability during gliding in flying snakes.

Science.gov (United States)

Jafari, Farid; Ross, Shane D; Vlachos, Pavlos P; Socha, John J

2014-06-01

Flying snakes use their entire body as a continuously morphing 'wing' to produce lift and shallow their glide trajectory. Their dominant behavior during gliding is aerial undulation, in which lateral waves are sent posteriorly down the body. This highly dynamic behavior, which is unique among animal gliders, should have substantial effects on the flight dynamics and stability of the snakes, resulting from the continuous redistribution of mass and aerodynamic forces. In this study, we develop two-dimensional theoretical models to assess the stability characteristics of snakes in the pitch direction. Previously measured force coefficients are used to simulate aerodynamic forces acting on the models, and undulation is simulated by varying mass. Model 1 is a simple three-airfoil representation of the snake's body that possesses a passively stable equilibrium solution, whose basin of stability contains initial conditions observed in experimental gliding trajectories. Model 2 is more sophisticated, with more degrees of freedom allowing for postural changes to better represent the snake's real kinematics; in addition, a restoring moment is added to simulate potential active control. The application of static and dynamic stability criteria show that Model 2 is passively unstable, but can be stabilized with a restoring moment. Overall, these models suggest that undulation does not contribute to stability in pitch, and that flying snakes require a closed-loop control system formed around a passively stable dynamical framework.

Plastic deformation of tubular crystals by dislocation glide.

Science.gov (United States)

Beller, Daniel A; Nelson, David R

2016-09-01

Tubular crystals, two-dimensional lattices wrapped into cylindrical topologies, arise in many contexts, including botany and biofilaments, and in physical systems such as carbon nanotubes. The geometrical principles of botanical phyllotaxis, describing the spiral packings on cylinders commonly found in nature, have found application in all these systems. Several recent studies have examined defects in tubular crystals associated with crystalline packings that must accommodate a fixed tube radius. Here we study the mechanics of tubular crystals with variable tube radius, with dislocations interposed between regions of different phyllotactic packings. Unbinding and separation of dislocation pairs with equal and opposite Burgers vectors allow the growth of one phyllotactic domain at the expense of another. In particular, glide separation of dislocations offers a low-energy mode for plastic deformations of solid tubes in response to external stresses, reconfiguring the lattice step by step. Through theory and simulation, we examine how the tube's radius and helicity affects, and is in turn altered by, the mechanics of dislocation glide. We also discuss how a sufficiently strong bending rigidity can alter or arrest the deformations of tubes with small radii.

Hypergravity synthesis of graphitic carbon nanomaterial in glide arc plasma

NARCIS (Netherlands)

Šperka, J.; Soucek, P.; van Loon, J.J.W.A.; Dowson, A.; Schwarz, C.; Krause, J.; Butenko, Y.; Kroesen, G.; Kudrle, V.

2014-01-01

A nanostructured carbon material was synthesized using a methane/helium glide arc plasma under standard and increased gravity. Material analysis performed on samples collected from an effluent gas filter showed that the deposited material was present in the form of carbon nanoparticles. They

Energy consumption of ProTaper Next X1 after glide path with PathFiles and ProGlider.

Science.gov (United States)

Berutti, Elio; Alovisi, Mario; Pastorelli, Michele Angelo; Chiandussi, Giorgio; Scotti, Nicola; Pasqualini, Damiano

2014-12-01

Instrument failure caused by excessive torsional stress can be controlled by creating a manual or mechanical glide path. The ProGlider single-file system (Dentsply Maillefer, Ballaigues, Switzerland) was recently introduced to perform a mechanical glide path. This study was designed to compare the effect of a glide path performed with PathFiles (Dentsply Maillefer) and ProGlider on torque, time, and pecking motion required for ProTaper Next X1 (Dentsply Maillefer) to reach the full working length in simulated root canals. Forty Endo Training Blocks (Dentsply Maillefer) were used. Twenty were prepared with a mechanical glide path using PathFiles 1 and 2 (the PathFile group), and 20 were prepared with a mechanical glide path using a ProGlider single file (the ProGlider group). All samples were shaped with ProTaper Next X1 driven by an endodontic motor connected to a digital wattmeter. The required torque for root canal instrumentation was analyzed by evaluating the electrical power consumption of the endodontic engine. Electric power consumption (mW/h), elapsed time (seconds), and number of pecking motions required to reach the full working length with ProTaper Next X1 were calculated. Differences among groups were analyzed with the parametric Student t test for independent data (P < .05). Elapsed time and electric power consumption were significantly different between groups (P = .0001 for both). ProGlider appears to perform more efficiently than PathFiles in decreasing electric power consumption of ProTaper Next X1 to reach the full working length. This study confirmed the ability of ProGlider to reduce stress in ProTaper Next X1 during shaping through a glide path and preliminary middle and coronal preflaring. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Perception of visual apparent motion is modulated by a gap within concurrent auditory glides, even when it is illusory

Science.gov (United States)

Wang, Qingcui; Guo, Lu; Bao, Ming; Chen, Lihan

2015-01-01

Auditory and visual events often happen concurrently, and how they group together can have a strong effect on what is perceived. We investigated whether/how intra- or cross-modal temporal grouping influenced the perceptual decision of otherwise ambiguous visual apparent motion. To achieve this, we juxtaposed auditory gap transfer illusion with visual Ternus display. The Ternus display involves a multi-element stimulus that can induce either of two different percepts of apparent motion: ‘element motion’ (EM) or ‘group motion’ (GM). In “EM,” the endmost disk is seen as moving back and forth while the middle disk at the central position remains stationary; while in “GM,” both disks appear to move laterally as a whole. The gap transfer illusion refers to the illusory subjective transfer of a short gap (around 100 ms) from the long glide to the short continuous glide when the two glides intercede at the temporal middle point. In our experiments, observers were required to make a perceptual discrimination of Ternus motion in the presence of concurrent auditory glides (with or without a gap inside). Results showed that a gap within a short glide imposed a remarkable effect on separating visual events, and led to a dominant perception of GM as well. The auditory configuration with gap transfer illusion triggered the same auditory capture effect. Further investigations showed that visual interval which coincided with the gap interval (50–230 ms) in the long glide was perceived to be shorter than that within both the short glide and the ‘gap-transfer’ auditory configurations in the same physical intervals (gaps). The results indicated that auditory temporal perceptual grouping takes priority over the cross-modal interaction in determining the final readout of the visual perception, and the mechanism of selective attention on auditory events also plays a role. PMID:26042055

Perception of visual apparent motion is modulated by a gap within concurrent auditory glides, even when it is illusory

Directory of Open Access Journals (Sweden)

Qingcui eWang

2015-05-01

Full Text Available Auditory and visual events often happen concurrently, and how they group together can have a strong effect on what is perceived. We investigated whether/how intra- or cross-modal temporal grouping influenced the perceptual decision of otherwise ambiguous visual apparent motion. To achieve this, we juxtaposed auditory gap transfer illusion with visual Ternus display. The Ternus display involves a multi-element stimulus that can induce either of two different percepts of apparent motion: ‘element motion’ or ‘group motion’. In element motion, the endmost disk is seen as moving back and forth while the middle disk at the central position remains stationary; while in group motion, both disks appear to move laterally as a whole. The gap transfer illusion refers to the illusory subjective transfer of a short gap (around 100 ms from the long glide to the short continuous glide when the two glides intercede at the temporal middle point. In our experiments, observers were required to make a perceptual discrimination of Ternus motion in the presence of concurrent auditory glides (with or without a gap inside. Results showed that a gap within a short glide imposed a remarkable effect on separating visual events, and led to a dominant perception of group motion as well. The auditory configuration with gap transfer illusion triggered the same auditory capture effect. Further investigations showed that visual interval which coincided with the gap interval (50-230 ms in the long glide was perceived to be shorter than that within both the short glide and the ‘gap-transfer’ auditory configurations in the same physical intervals (gaps. The results indicated that auditory temporal perceptual grouping takes priority over the cross-modal interaction in determining the final readout of the visual perception, and the mechanism of selective attention on auditory events also plays a role.

How Informative are the Vertical Buoyancy and the Prone Gliding Tests to Assess Young Swimmers’ Hydrostatic and Hydrodynamic Profiles?

Science.gov (United States)

Barbosa, Tiago M.; Costa, Mário J.; Morais, Jorge E; Moreira, Marc; Silva, António J.; Marinho, Daniel A.

2012-01-01

The aim of this research was to develop a path-flow analysis model to highlight the relationships between buoyancy and prone gliding tests and some selected anthropometrical and biomechanical variables. Thirty-eight young male swimmers (12.97 ± 1.05 years old) with several competitive levels were evaluated. It were assessed the body mass, height, fat mass, body surface area, vertical buoyancy, prone gliding after wall push-off, stroke length, stroke frequency and velocity after a maximal 25 [m] swim. The confirmatory model included the body mass, height, fat mass, prone gliding test, stroke length, stroke frequency and velocity. All theoretical paths were verified except for the vertical buoyancy test that did not present any relationship with anthropometrical and biomechanical variables nor with the prone gliding test. The good-of-fit from the confirmatory path-flow model, assessed with the standardized root mean square residuals (SRMR), is considered as being close to the cut-off value, but even so not suitable of the theory (SRMR = 0.11). As a conclusion, vertical buoyancy and prone gliding tests are not the best techniques to assess the swimmer’s hydrostatic and hydrodynamic profile, respectively. PMID:23486528

Marked colour divergence in the gliding membranes of a tropical lizard mirrors population differences in the colour of falling leaves

OpenAIRE

Klomp, D. A.; Stuart-Fox, D.; Das, I.; Ord, T. J.

2014-01-01

Populations of the Bornean gliding lizard, Draco cornutus, differ markedly in the colour of their gliding membranes. They also differ in local vegetation type (mangrove forest versus lowland rainforest) and consequently, the colour of falling leaves (red and brown/black in mangrove versus green, brown and black in rainforest). We show that the gliding membranes of these lizards closely match the colours of freshly fallen leaves in the local habitat as they appear to the visual system of birds...

Kick, Glide, Pole! Cross-Country Skiing Fun (Part II)

Science.gov (United States)

Duoos, Bridget A.

2012-01-01

Part I of Kick, Glide, Pole! Cross-Country Skiing Fun, which was published in last issue, discussed how to select cross-country ski equipment, dress for the activity and the biomechanics of the diagonal stride. Part II focuses on teaching the diagonal stride technique and begins with a progression of indoor activities. Incorporating this fun,…

Investigation of Aerodynamic Capabilities of Flying Fish in Gliding Flight

Science.gov (United States)

Park, H.; Choi, H.

In the present study, we experimentally investigate the aerodynamic capabilities of flying fish. We consider four different flying fish models, which are darkedged-wing flying fishes stuffed in actual gliding posture. Some morphological parameters of flying fish such as lateral dihedral angle of pectoral fins, incidence angles of pectoral and pelvic fins are considered to examine their effect on the aerodynamic performance. We directly measure the aerodynamic properties (lift, drag, and pitching moment) for different morphological parameters of flying fish models. For the present flying fish models, the maximum lift coefficient and lift-to-drag ratio are similar to those of medium-sized birds such as the vulture, nighthawk and petrel. The pectoral fins are found to enhance the lift-to-drag ratio and the longitudinal static stability of gliding flight. On the other hand, the lift coefficient and lift-to-drag ratio decrease with increasing lateral dihedral angle of pectoral fins.

Evidence and characterization of a glide-vowel distinction in American English

Directory of Open Access Journals (Sweden)

Zachary Scott Jaggers

2018-02-01

Full Text Available This study tests whether native speakers of American English exhibit a glide-vowel distinction ([j]-[i] in a speech elicitation experiment. When reading sentences out loud, participants’ pronunciations of 4 near-minimal pairs of pre-existing lexical items (e.g., 'Eston'[iə] vs. 'pneumon'[jə] exhibit significant differences when acoustically measured, confirming the presence of a [j]-[i] distinction. This distinction is also found to be productively extended to the production of 20 near-minimal pairs of nonce words (e.g., 'Súmia '→ [sumiə] vs. 'Fímya '→ [fimjə], diversified and balanced along different phonologically relevant factors of the surrounding environment. Multiple acoustic measurements are compared to test what aspects most consistently convey the distinction: F2 (frontness, F1 (height, intensity, vocalic sequence duration, transition earliness, and transition speed. This serves the purpose of documenting the distinction’s acoustic phonetic realization. It also serves in the comparison of phonological representations. Multiple types of previously proposed phonological representations are considered along with the competing predictions they generate regarding the acoustic measurements performed. Results suggest that the primary and most consistent characteristic of the distinction is earliness of transition into the following vowel, with results also suggesting that the [j] glide has a greater degree of constriction. The [j] glide is found to have a significantly 'less 'anterior articulation, challenging the application of a representation based on place or articulator differences that would predict [j] to be 'more 'anterior.

A theoretical analysis of pitch stability during gliding in flying snakes

International Nuclear Information System (INIS)

Jafari, Farid; Ross, Shane D; Socha, John J; Vlachos, Pavlos P

2014-01-01

Flying snakes use their entire body as a continuously morphing ‘wing’ to produce lift and shallow their glide trajectory. Their dominant behavior during gliding is aerial undulation, in which lateral waves are sent posteriorly down the body. This highly dynamic behavior, which is unique among animal gliders, should have substantial effects on the flight dynamics and stability of the snakes, resulting from the continuous redistribution of mass and aerodynamic forces. In this study, we develop two-dimensional theoretical models to assess the stability characteristics of snakes in the pitch direction. Previously measured force coefficients are used to simulate aerodynamic forces acting on the models, and undulation is simulated by varying mass. Model 1 is a simple three-airfoil representation of the snake’s body that possesses a passively stable equilibrium solution, whose basin of stability contains initial conditions observed in experimental gliding trajectories. Model 2 is more sophisticated, with more degrees of freedom allowing for postural changes to better represent the snake’s real kinematics; in addition, a restoring moment is added to simulate potential active control. The application of static and dynamic stability criteria show that Model 2 is passively unstable, but can be stabilized with a restoring moment. Overall, these models suggest that undulation does not contribute to stability in pitch, and that flying snakes require a closed-loop control system formed around a passively stable dynamical framework. (papers)

Ultrasound-guided hydrodissection decreases gliding resistance of the median nerve within the carpal tunnel.

Science.gov (United States)

Evers, Stefanie; Thoreson, Andrew R; Smith, Jay; Zhao, Chunfeng; Geske, Jennifer R; Amadio, Peter C

2018-01-01

The aim of this study was to assess alterations in median nerve (MN) biomechanics within the carpal tunnel resulting from ultrasound-guided hydrodissection in a cadaveric model. Twelve fresh frozen human cadaver hands were used. MN gliding resistance was measured at baseline and posthydrodissection, by pulling the nerve proximally and then returning it to the origin. Six specimens were treated with hydrodissection, and 6 were used as controls. In the hydrodissection group there was a significant reduction in mean peak gliding resistance of 92.9 ± 34.8 mN between baseline and immediately posthydrodissection (21.4% ± 10.5%; P = 0.001). No significant reduction between baseline and the second cycle occurred in the control group: 9.6 ± 29.8 mN (0.4% ± 5.3%; P = 0.467). Hydrodissection can decrease the gliding resistance of the MN within the carpal tunnel, at least in wrists unaffected by carpal tunnel syndrome. A clinical trial of hydrodissection seems justified. Muscle Nerve 57: 25-32, 2018. © 2017 Wiley Periodicals, Inc.

Estimating intratidal nonlinearity of respiratory system mechanics: a model study using the enhanced gliding-SLICE method

International Nuclear Information System (INIS)

Schumann, Stefan; Burcza, Boris; Guttmann, Josef; Haberthür, Christoph; Lichtwarck-Aschoff, Michael

2009-01-01

In the clinical situation and in most research work, the analysis of respiratory system mechanics is limited to the estimation of single-value compliances during static or quasi-static conditions. In contrast, our SLICE method analyses intratidal nonlinearity under the dynamic conditions of mechanical ventilation by calculating compliance and resistance for six conjoined volume portions (slices) of the pressure–volume loop by multiple linear regression analysis. With the gliding-SLICE method we present a new approach to determine continuous intratidal nonlinear compliance. The performance of the gliding-SLICE method was tested both in computer simulations and in a physical model of the lung, both simulating different intratidal compliance profiles. Compared to the original SLICE method, the gliding-SLICE method resulted in smaller errors when calculating the compliance or pressure course (all p 2 O s L −1 to 0.8 ± 0.3 cmH 2 O s L −1 (mathematical model) and from 7.2 ± 3.9 cmH 2 O s L −1 to 0.4 ± 0.2 cmH 2 O s L −1 (physical model) (all p < 0.001). We conclude that the new gliding-SLICE method allows detailed assessment of intratidal nonlinear respiratory system mechanics without discontinuity error

Degradation of Verapamil hydrochloride in water by gliding arc discharge

Czech Academy of Sciences Publication Activity Database

Krishna, S.; Mašláni, Alan; Izdebski, T.; Horáková, M.; Klementová, Š.; Špatenka, P.

2016-01-01

Roč. 152, June (2016), s. 47-54 ISSN 0045-6535 Institutional support: RVO:61389021 Keywords : Gliding arc discharge * Emission spectroscopy * Pharmaceuticals * Half-life * Degradation mechanism Subject RIV: BL - Plasma and Gas Discharge Physics Impact factor: 4.208, year: 2016 http://www.sciencedirect.com/science/article/pii/S0045653516302442

A rolling-gliding wear simulator for the investigation of tribological material pairings for application in total knee arthroplasty

Science.gov (United States)

2010-01-01

Background Material wear testing is an important technique in the development and evaluation of materials for use in implant for total knee arthroplasty. Since a knee joint induces a complex rolling-gliding movement, standardised material wear testing devices such as Pin-on-Disc or Ring-on-Disc testers are suitable to only a limited extent because they generate pure gliding motion only. Methods A rolling-gliding wear simulator was thus designed, constructed and implemented, which simulates and reproduces the rolling-gliding movement and loading of the knee joint on specimens of simplified geometry. The technical concept was to run a base-plate, representing the tibia plateau, against a pivoted cylindrical counter-body, representing one femur condyle under an axial load. A rolling movement occurs as a result of the friction and pure gliding is induced by limiting the rotation of the cylindrical counter-body. The set up also enables simplified specimens handling and removal for gravimetrical wear measurements. Long-term wear tests and gravimetrical wear measurements were carried out on the well known material pairings: cobalt chrome-polyethylene, ceramic-polyethylene and ceramic-ceramic, over three million motion cycles to allow material comparisons to be made. Results The observed differences in wear rates between cobalt-chrome on polyethylene and ceramic on polyethylene pairings were similar to the differences of published data for existing material-pairings. Test results on ceramic-ceramic pairings of different frontal-plane geometry and surface roughness displayed low wear rates and no fracture failures. Conclusions The presented set up is able to simulate the rolling-gliding movement of the knee joint, is easy to use, and requires a minimum of user intervention or monitoring. It is suitable for long-term testing, and therefore a useful tool for the investigation of new and promising materials which are of interest for application in knee joint replacement

A rolling-gliding wear simulator for the investigation of tribological material pairings for application in total knee arthroplasty

Directory of Open Access Journals (Sweden)

Denkena Berend

2010-06-01

Full Text Available Abstract Background Material wear testing is an important technique in the development and evaluation of materials for use in implant for total knee arthroplasty. Since a knee joint induces a complex rolling-gliding movement, standardised material wear testing devices such as Pin-on-Disc or Ring-on-Disc testers are suitable to only a limited extent because they generate pure gliding motion only. Methods A rolling-gliding wear simulator was thus designed, constructed and implemented, which simulates and reproduces the rolling-gliding movement and loading of the knee joint on specimens of simplified geometry. The technical concept was to run a base-plate, representing the tibia plateau, against a pivoted cylindrical counter-body, representing one femur condyle under an axial load. A rolling movement occurs as a result of the friction and pure gliding is induced by limiting the rotation of the cylindrical counter-body. The set up also enables simplified specimens handling and removal for gravimetrical wear measurements. Long-term wear tests and gravimetrical wear measurements were carried out on the well known material pairings: cobalt chrome-polyethylene, ceramic-polyethylene and ceramic-ceramic, over three million motion cycles to allow material comparisons to be made. Results The observed differences in wear rates between cobalt-chrome on polyethylene and ceramic on polyethylene pairings were similar to the differences of published data for existing material-pairings. Test results on ceramic-ceramic pairings of different frontal-plane geometry and surface roughness displayed low wear rates and no fracture failures. Conclusions The presented set up is able to simulate the rolling-gliding movement of the knee joint, is easy to use, and requires a minimum of user intervention or monitoring. It is suitable for long-term testing, and therefore a useful tool for the investigation of new and promising materials which are of interest for application in

A rolling-gliding wear simulator for the investigation of tribological material pairings for application in total knee arthroplasty.

Science.gov (United States)

Richter, Berna I; Ostermeier, Sven; Turger, Anke; Denkena, Berend; Hurschler, Christof

2010-06-15

Material wear testing is an important technique in the development and evaluation of materials for use in implant for total knee arthroplasty. Since a knee joint induces a complex rolling-gliding movement, standardised material wear testing devices such as Pin-on-Disc or Ring-on-Disc testers are suitable to only a limited extent because they generate pure gliding motion only. A rolling-gliding wear simulator was thus designed, constructed and implemented, which simulates and reproduces the rolling-gliding movement and loading of the knee joint on specimens of simplified geometry. The technical concept was to run a base-plate, representing the tibia plateau, against a pivoted cylindrical counter-body, representing one femur condyle under an axial load. A rolling movement occurs as a result of the friction and pure gliding is induced by limiting the rotation of the cylindrical counter-body. The set up also enables simplified specimens handling and removal for gravimetrical wear measurements. Long-term wear tests and gravimetrical wear measurements were carried out on the well known material pairings: cobalt chrome-polyethylene, ceramic-polyethylene and ceramic-ceramic, over three million motion cycles to allow material comparisons to be made. The observed differences in wear rates between cobalt-chrome on polyethylene and ceramic on polyethylene pairings were similar to the differences of published data for existing material-pairings. Test results on ceramic-ceramic pairings of different frontal-plane geometry and surface roughness displayed low wear rates and no fracture failures. The presented set up is able to simulate the rolling-gliding movement of the knee joint, is easy to use, and requires a minimum of user intervention or monitoring. It is suitable for long-term testing, and therefore a useful tool for the investigation of new and promising materials which are of interest for application in knee joint replacement implants.

Optimization geometries of a vortex gliding-arc reactor for partial oxidation of methane

International Nuclear Information System (INIS)

Guofeng, Xu; Xinwei, Ding

2012-01-01

The effects of the geometry of gliding-arc reactor – such as distance between the electrodes, outlet diameter, and inlet position – on the reactor characteristics (methane conversion, hydrogen yield, and energy efficiency) have not been fully investigated. In this paper, AC gliding-arc reactors including the vortex flow configuration are designed to produce hydrogen from the methane by partial oxidation. The influence of vortex flow configuration on the reactor characteristics is also studied by varying the inlet position. When the inlet of the gliding-arc reactor is positioned close to the outlet, reverse vortex flow reactor (RVFR), the maximum energy efficiency reaches 50% and the yields of hydrogen and carbon monoxide are 40% and 65%, respectively. As the distance between electrodes increases from 5 mm to 15 mm, both hydrogen yield and energy efficiency increase approximately 10% for the RVFR. The energy efficiency and hydrogen yield are highest when the ratio of the outlet diameter to the inner diameter is 0.5 for the RVFR. Experimental results indicate that the flow field in the plasma reactor has an important influence on the reactor performance. Furthermore, hydrogen production increases as the number of feed gas flows in contact with the plasma zone increases. -- Highlights: ► Gliding-arc reactors were designed to produce hydrogen for studying the characteristics of the vortex flow reactor. ► Hydrogen yield of reverse vortex flow reactor was 10% higher than that of forward vortex flow reactor. ► Maximum energy efficiency was 50% for reverse vortex flow reactor. ► If discharge power was supplied to the reactors, the reactor performance increased with increasing distance between electrodes. ► Optimum ratio of the outlet and inner diameter was 1/2.

The Hydrodynamic Study of the Swimming Gliding: a Two-Dimensional Computational Fluid Dynamics (CFD) Analysis.

Science.gov (United States)

Marinho, Daniel A; Barbosa, Tiago M; Rouboa, Abel I; Silva, António J

2011-09-01

Nowadays the underwater gliding after the starts and the turns plays a major role in the overall swimming performance. Hence, minimizing hydrodynamic drag during the underwater phases should be a main aim during swimming. Indeed, there are several postures that swimmers can assume during the underwater gliding, although experimental results were not conclusive concerning the best body position to accomplish this aim. Therefore, the purpose of this study was to analyse the effect in hydrodynamic drag forces of using different body positions during gliding through computational fluid dynamics (CFD) methodology. For this purpose, two-dimensional models of the human body in steady flow conditions were studied. Two-dimensional virtual models had been created: (i) a prone position with the arms extended at the front of the body; (ii) a prone position with the arms placed alongside the trunk; (iii) a lateral position with the arms extended at the front and; (iv) a dorsal position with the arms extended at the front. The drag forces were computed between speeds of 1.6 m/s and 2 m/s in a two-dimensional Fluent(®) analysis. The positions with the arms extended at the front presented lower drag values than the position with the arms aside the trunk. The lateral position was the one in which the drag was lower and seems to be the one that should be adopted during the gliding after starts and turns.

A Modified NK Cell Degranulation Assay Applicable for Routine Evaluation of NK Cell Function

Directory of Open Access Journals (Sweden)

Snehal Shabrish

2016-01-01

Full Text Available Natural killer (NK cells play important role in innate immunity against tumors and viral infections. Studies show that lysosome-associated membrane protein-1 (LAMP-1, CD107a is a marker for degranulation of NK and cytotoxic T cells and its expression is a sensitive marker for the cytotoxic activity determination. The conventional methods of determination of CD107a on NK cells involve use of peripheral blood mononuclear cells (PBMC or pure NK cells and K562 cells as stimulants. Thus, it requires large volume of blood sample which is usually difficult to obtain in pediatric patients and patients with cytopenia and also requires specialized laboratory for maintaining cell line. We have designed a flow cytometric assay to determine CD107a on NK cells using whole blood, eliminating the need for isolation of PBMC or isolate NK cells. This assay uses phorbol-12-myristate-13-acetate (PMA and calcium ionophore (Ca2+-ionophore instead of K562 cells for stimulation and thus does not require specialized cell culture laboratory. CD107a expression on NK cells using modified NK cell degranulation assay compared to the conventional assay was significantly elevated (p<0.0001. It was also validated by testing patients diagnosed with familial hemophagocytic lymphohistiocytosis (FHL with defect in exocytosis. This assay is rapid, cost effective, and reproducible and requires significantly less volume of blood which is important for clinical evaluation of NK cells.

Sustained diffusive alternating current gliding arc discharge in atmospheric pressure air

Science.gov (United States)

Zhu, Jiajian; Gao, Jinlong; Li, Zhongshan; Ehn, Andreas; Aldén, Marcus; Larsson, Anders; Kusano, Yukihiro

2014-12-01

Rapid transition from glow discharge to thermal arc has been a common problem in generating stable high-power non-thermal plasmas especially at ambient conditions. A sustained diffusive gliding arc discharge was generated in a large volume in atmospheric pressure air, driven by an alternating current (AC) power source. The plasma column extended beyond the water-cooled stainless steel electrodes and was stabilized by matching the flow speed of the turbulent air jet with the rated output power. Comprehensive investigations were performed using high-speed movies measured over the plasma column, synchronized with simultaneously recorded current and voltage waveforms. Dynamic details of the novel non-equilibrium discharge are revealed, which is characterized by a sinusoidal current waveform with amplitude stabilized at around 200 mA intermediate between thermal arc and glow discharge, shedding light to the governing mechanism of the sustained spark-suppressed AC gliding arc discharge.

On low temperature glide of dissociated 〈1 1 0〉 dislocations in strontium titanate

Science.gov (United States)

Ritterbex, Sebastian; Hirel, Pierre; Carrez, Philippe

2018-05-01

An elastic interaction model is presented to quantify low temperature plasticity of SrTiO3 via glide of dissociated 〈1 1 0〉{1 1 0} screw dislocations. Because 〈1 1 0〉 dislocations are dissociated, their glide, controlled by the kink-pair mechanism at T good quantitative agreement with the observed non-monotonic mechanical behaviour of SrTiO3. This agreement allows to explain the experimental results in terms of a (progressive) change in 〈1 1 0〉{1 1 0} glide mechanism, from simultaneous nucleation of two kink-pairs along both partials at low stress, towards nucleation of single kink-pairs on individual partials if resolved shear stress exceeds a critical value of 95 MPa. High resolved shear stress allows thus for the activation of extra nucleation mechanisms on dissociated dislocations impossible to occur under the sole action of thermal activation. We suggest that stress condition in conjunction with core dissociation is key to the origin of non-monotonic plastic behaviour of SrTiO3 at low temperatures.

Degradation of gas-liquid gliding arc discharge on Acid Orange II

International Nuclear Information System (INIS)

Yan, J.H.; Liu, Y.N.; Bo, Zh.; Li, X.D.; Cen, K.F.

2008-01-01

The effects of pH value, initial concentration of dye solution and temperature on the degradation efficiency of Acid Orange II (AO7) using gas-liquid gliding arc discharge were investigated. The influences of pH value and temperature on degradation efficiency were not apparent. Increasing initial solution concentration caused the decrease of degradation rate and the increase of absolute degradation quantity. Considering energy efficiency and absolute degradation quantity, the gas-liquid gliding arc discharge is fit for treating high concentration organic wastewater. A possible mineralization pathway was proposed through the analysis of intermediate products detected by gas chromatograph coupled with mass spectrophotometer (GC-MS) and ion chromatograph (IC). Hydroxyl radicals reacted with the azo linkage-bearing carbon of a hydroxy-substituted ring, leading to the cleavage of -C-N- and degradation of AO7. The solution biodegradability was significantly improved (BOD 5 /COD from 0.02 to 0.43). The toxicity of intermediate products was lower than that of the initial Acid Orange II

Pulsating-gliding transition in the dynamics of levitating liquid nitrogen droplets

International Nuclear Information System (INIS)

Snezhko, Alexey; Aranson, Igor S; Jacob, Eshel Ben

2008-01-01

Hot surfaces can cause levitation of small liquid droplets if the temperature is kept above the Leidenfrost point (220 0 C for water) due to the pressure formed because of rapid evaporation. Here, we demonstrate a new class of pulsating-gliding dynamic transitions in a special setting of the Leidenfrost effect at room temperatures and above a viscous fluid for droplets of liquid nitrogen. A whole range of highly dynamic patterns unfolds when droplets of liquid nitrogen are poured on the surface of another, more viscous liquid at room temperature. We also discovered that the levitating droplets induce vortex motion in the supporting viscous liquid. Depending on the viscosity of the supporting liquid, the nitrogen droplets either adopt an oscillating (pulsating) star-like shape with different azimuthal symmetries (from 2-9 petals) or glide on the surface with random trajectories. Thus, by varying the viscosity of the supporting liquid, we achieve controlled morphology and dynamics of Leidenfrost droplets

Pulsating-gliding transition in the dynamics of levitating liquid nitrogen droplets

Energy Technology Data Exchange (ETDEWEB)

Snezhko, Alexey; Aranson, Igor S [Materials Science Division, Argonne National Laboratory, 9700 S Cass Avenue, Argonne, IL 60439 (United States); Jacob, Eshel Ben [School of Physics and Astronomy, 69978 Tel Aviv University, Tel Aviv (Israel)], E-mail: aranson@msd.anl.gov

2008-04-15

Hot surfaces can cause levitation of small liquid droplets if the temperature is kept above the Leidenfrost point (220 {sup 0}C for water) due to the pressure formed because of rapid evaporation. Here, we demonstrate a new class of pulsating-gliding dynamic transitions in a special setting of the Leidenfrost effect at room temperatures and above a viscous fluid for droplets of liquid nitrogen. A whole range of highly dynamic patterns unfolds when droplets of liquid nitrogen are poured on the surface of another, more viscous liquid at room temperature. We also discovered that the levitating droplets induce vortex motion in the supporting viscous liquid. Depending on the viscosity of the supporting liquid, the nitrogen droplets either adopt an oscillating (pulsating) star-like shape with different azimuthal symmetries (from 2-9 petals) or glide on the surface with random trajectories. Thus, by varying the viscosity of the supporting liquid, we achieve controlled morphology and dynamics of Leidenfrost droplets.

A mannequin study of intubation with the AP advance and GlideScope Ranger videolaryngoscopes and the Macintosh laryngoscope.

Science.gov (United States)

Hodd, Jack A R; Doyle, D John; Gupta, Shipra; Dalton, Jarrod E; Cata, Juan P; Brewer, Edward J; James, Monyulona; Sessler, Daniel I

2011-10-01

The AP Advance (APA) is a videolaryngoscope with interchangeable blades: intubators can choose standard Macintosh blades or a difficult-airway blade with increased curvature and a channel to guide the tube to the larynx. The APA may therefore be comparably effective in both normal and difficult airways. We tested the hypotheses that intubation with the APA is no slower than Macintosh laryngoscopy for normal mannequin airways, and that it is no slower than videolaryngoscopy using a GlideScope Ranger in difficult mannequin airways. Medical professionals whose roles potentially include tracheal intubation were trained with each device. Participants intubated simulated (Laerdal SimMan) normal and difficult airways with the APA, GlideScope, and a conventional Macintosh blade. Speed of intubation was compared using Cox proportional hazards regression, with a hazard ratio >0.8 considered noninferior. We also compared laryngeal visualization, failures, and participant preferences. Unadjusted intubation times in the normal airway with the APA and Macintosh were virtually identical (median, 22 vs 23 seconds); after adjustment for effects of experience, order, and period, the hazard ratio (95% confidence interval) comparing APA with Macintosh laryngoscopy was 0.87 (0.65, 1.17), which was not significantly more than our predefined noninferiority boundary of 0.8 (P = 0.26). Intubation with the APA was faster than with the GlideScope in difficult airways (hazard ratio = 7.6 [5.0, 11.3], P APA, whereas 33% and 37% failed with the GlideScope and Macintosh, respectively. In the difficult airway, 99% of participants achieved a Cormack and Lehane grade I to II view with the APA, versus 85% and 33% with the GlideScope and Macintosh, respectively. When asked to choose 1 device overall, 82% chose the APA. Intubation times were similar with the APA and Macintosh laryngoscopes in mannequins with normal airways. However, intubation with the APA was significantly faster than with the Glide

An in vitro comparison of root canal transportation by reciproc file with and without glide path.

Science.gov (United States)

Nazarimoghadam, Kiumars; Daryaeian, Mohammad; Ramazani, Nahid

2014-09-01

The aim of ideal canal preparation is to prevent iatrogenic aberrations such as transportation. The aim of this study was to evaluate the root canal transportation by Reciproc file with and without glide path. Thirty acrylic-resin blocks with a curvature of 60° and size#10 (2% taper) were assigned into two groups (n= 15). In group 1, the glide path was performed using stainless steel k-files size#10 and 15 at working length In group 2, canals were prepared with Reciproc file system at working length. By using digital imaging software (AutoCAD 2008), the pre-instrumentation and post-instrumentation digital images were superimposed over, taking the landmarks as reference points. Then the radius of the internal and external curve of the specimens was calculated at three α, β and γ points (1mm to apex as α, 3mm to apex as β, and 5mm to apex as γ). The data were statically analyzed using the independent T-test and Mann-Whitney U test by SPSS version 16. Glide path was found significant for only external curve in the apical third of the canal; that is, 5mm to apex (P=0.005). But in the other third, canal modification was not significant (P> 0.008). Canal transportation in the apical third of the canal seems to be significantly reduced when glide path is performed using reciprocating files.

Direct observation of gliding dislocations interactions with defects in irradiated niobium single crystals by means of the high voltage electronic microscopy (HVEM)

International Nuclear Information System (INIS)

Otero, M.P.

1985-01-01

The interactions of gliding dislocations with defects in irradiated niobium that result in the formation of dislocations channels. The effects in the mechanical behaviour of [941]- and [441]- oriented Nb single crystals due to oxygen addition, neutron and electron irradiation was observed either by macroscopic deformation in a Instron machine or 'in-situ' deformation in the HVEM-High Voltage Electron Microscope. Some specimens were irradiated at IPNS-Intense Pulsed Neutron Source, at 325 K, with 5 x 10 17 n/cm 2 , others were irradiated with electrons in the HVEM. The interactions between gliding dislocations with clusters point defects and dislocations were observed. The primary mechanism for removal of the clusters by the gliding dislocations was the 'sweeping' of the clusters along with the gliding dislocations. As to the point defects, they were 'swept' by the gliding dislocations and left as aligned loops close to the intersections of the gliding dislocations with the upper and lower specimen surfaces. For the illustration of this phenomena, a schematic drawing was made. The mechanism of 'bowing-out' interaction of dislocations with defect clusters was also observed. The reported anomalous slip observed to operate in the [941]- oriented Nb was also directly observed and a qualitive explanation along with a schematic drawing was proposed. This would explain the softenig observed after the yield stress in the [941]- oriented Nb deformed in the Instron machine. (Author) [pt

Destruction of acenaphthene, fluorene, anthracene and pyrene by a dc gliding arc plasma reactor

International Nuclear Information System (INIS)

Yu Liang; Tu Xin; Li Xiaodong; Wang Yu; Chi Yong; Yan Jianhua

2010-01-01

In this study, four kinds of PAHs (polycyclic aromatic hydrocarbons) i.e. acenaphthene, fluorene, anthracene and pyrene are used as targets for investigation of PAHs treatment process assisted by dc gliding arc discharge. The effects of carrier gas and external resistance on the PAHs decomposition process are discussed. The results indicate that the destruction rate can be achieved to the highest with the carrier gas of oxygen and the external resistance of 50 kΩ independent of type of PAHs. Furthermore, experimental results suggest that destruction energy efficiency of gliding arc plasma would be improved by treating higher concentration pollutants. Based on the analysis of experimental results, possible destruction mechanisms in different gas discharge are discussed.

Gliding arc in tornado using a reverse vortex flow

International Nuclear Information System (INIS)

Kalra, Chiranjeev S.; Cho, Young I.; Gutsol, Alexander; Fridman, Alexander; Rufael, Tecle S.

2005-01-01

The present article reports a new gliding arc (GA) system using a reverse vortex flow ('tornado') in a cylindrical reactor (gliding arc in tornado, or GAT), as used to preserve the main advantages of traditional GA systems and overcome their main drawbacks. The primary advantages of traditional GA systems retained in the present GAT are the possibility to generate transitional plasma and to avoid considerable electrode erosion. In contrast to a traditional GA, the new GAT system ensures much more uniform gas treatment and has a significantly larger gas residence time in the reactor. The present article also describes the design of the new reactor and its stable operation regime when the variation of GAT current is very small. These features are understood to be very important for most viable applications. Additionally the GAT provides near-perfect thermal insulation from the reactor wall, indicating that the present GAT does not require the reactor wall to be constructed of high-temperature materials. The new GAT system, with its unique properties such as a high level of nonequilibrium and a large residence time, looks very promising for many industrial applications including fuel conversion, carbon dioxide conversion to carbon monoxide and oxygen, surface treatment, waste treatment, flame stabilization, hydrogen sulfide treatment, etc

Torsional Performance of ProTaper Gold Rotary Instruments during Shaping of Small Root Canals after 2 Different Glide Path Preparations.

Science.gov (United States)

Arias, Ana; de Vasconcelos, Rafaela Andrade; Hernández, Alexis; Peters, Ove A

2017-03-01

The purpose of this study was to assess the ex vivo torsional performance of a novel rotary system in small root canals after 2 different glide path preparations. Each independent canal of 8 mesial roots of mandibular molars was randomly assigned to achieve a reproducible glide path with a new set of either PathFile #1 (Dentsply Maillefer, Ballaigues, Switzerland) and #2 or ProGlider (Dentsply Maillefer) after negotiation with a 10 K-file. After glide path preparation, root canals in both groups were shaped with the same sequence of ProTaper Gold (Dentsply Tulsa Dental Specialties, Tulsa, OK) following the directions for use recommended by the manufacturer. A total of 16 new sets of each instrument of the ProTaper Gold (PTG) system were used. The tests were run in a standardized fashion in a torque-testing platform. Peak torque (Ncm) and force (N) were registered during the shaping procedure and compared with Student t tests after normal distribution of data was confirmed. No significant differences were found for any of the instruments in peak torque or force after the 2 different glide path preparations (P > .05). Data presented in this study also serve as a basis for the recommended torque for the use of PTG instruments. Under the conditions of this study, differences in the torsional performance of PTG rotary instruments after 2 different glide path preparations could not be shown. The different geometry of glide path rotary systems seemed to have no effect on peak torque and force induced by PTG rotary instruments when shaping small root canals in extracted teeth. Copyright © 2016 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Collaborative ring trial of the papaya endogenous reference gene and its polymerase chain reaction assays for genetically modified organism analysis.

Science.gov (United States)

Wei, Jiaojun; Li, Feiwu; Guo, Jinchao; Li, Xiang; Xu, Junfeng; Wu, Gang; Zhang, Dabing; Yang, Litao

2013-11-27

The papaya (Carica papaya L.) Chymopapain (CHY) gene has been reported as a suitable endogenous reference gene for genetically modified (GM) papaya detection in previous studies. Herein, we further validated the use of the CHY gene and its qualitative and quantitative polymerase chain reaction (PCR) assays through an interlaboratory collaborative ring trial. A total of 12 laboratories working on detection of genetically modified organisms participated in the ring trial and returned test results. Statistical analysis of the returned results confirmed the species specificity, low heterogeneity, and single-copy number of the CHY gene among different papaya varieties. The limit of detection of the CHY qualitative PCR assay was 0.1%, while the limit of quantification of the quantitative PCR assay was ∼25 copies of haploid papaya genome with acceptable PCR efficiency and linearity. The differences between the tested and true values of papaya content in 10 blind samples ranged from 0.84 to 6.58%. These results indicated that the CHY gene was suitable as an endogenous reference gene for the identification and quantification of GM papaya.

Differential recognition of pitch patterns in discrete and gliding stimuli in congenital amusia: evidence from Mandarin speakers.

Science.gov (United States)

Liu, Fang; Xu, Yi; Patel, Aniruddh D; Francart, Tom; Jiang, Cunmei

2012-08-01

This study examined whether "melodic contour deafness" (insensitivity to the direction of pitch movement) in congenital amusia is associated with specific types of pitch patterns (discrete versus gliding pitches) or stimulus types (speech syllables versus complex tones). Thresholds for identification of pitch direction were obtained using discrete or gliding pitches in the syllable /ma/ or its complex tone analog, from nineteen amusics and nineteen controls, all healthy university students with Mandarin Chinese as their native language. Amusics, unlike controls, had more difficulty recognizing pitch direction in discrete than in gliding pitches, for both speech and non-speech stimuli. Also, amusic thresholds were not significantly affected by stimulus types (speech versus non-speech), whereas controls showed lower thresholds for tones than for speech. These findings help explain why amusics have greater difficulty with discrete musical pitch perception than with speech perception, in which continuously changing pitch movements are prevalent. Copyright © 2012 Elsevier Inc. All rights reserved.

Destruction of acenaphthene, fluorene, anthracene and pyrene by a dc gliding arc plasma reactor.

Science.gov (United States)

Yu, Liang; Tu, Xin; Li, Xiaodong; Wang, Yu; Chi, Yong; Yan, Jianhua

2010-08-15

In this study, four kinds of PAHs (polycyclic aromatic hydrocarbons) i.e. acenaphthene, fluorene, anthracene and pyrene are used as targets for investigation of PAHs treatment process assisted by dc gliding arc discharge. The effects of carrier gas and external resistance on the PAHs decomposition process are discussed. The results indicate that the destruction rate can be achieved to the highest with the carrier gas of oxygen and the external resistance of 50 kOmega independent of type of PAHs. Furthermore, experimental results suggest that destruction energy efficiency of gliding arc plasma would be improved by treating higher concentration pollutants. Based on the analysis of experimental results, possible destruction mechanisms in different gas discharge are discussed. Copyright 2010 Elsevier B.V. All rights reserved.

A protein secretion system linked to bacteroidete gliding motility and pathogenesis

Science.gov (United States)

Sato, Keiko; Naito, Mariko; Yukitake, Hideharu; Hirakawa, Hideki; Shoji, Mikio; McBride, Mark J.; Rhodes, Ryan G.; Nakayama, Koji

2009-01-01

Porphyromonas gingivalis secretes strong proteases called gingipains that are implicated in periodontal pathogenesis. Protein secretion systems common to other Gram-negative bacteria are lacking in P. gingivalis, but several proteins, including PorT, have been linked to gingipain secretion. Comparative genome analysis and genetic experiments revealed 11 additional proteins involved in gingipain secretion. Six of these (PorK, PorL, PorM, PorN, PorW, and Sov) were similar in sequence to Flavobacterium johnsoniae gliding motility proteins, and two others (PorX and PorY) were putative two-component system regulatory proteins. Real-time RT-PCR analysis revealed that porK, porL, porM, porN, porP, porT, and sov were down-regulated in P. gingivalis porX and porY mutants. Disruption of the F. johnsoniae porT ortholog resulted in defects in motility, chitinase secretion, and translocation of a gliding motility protein, SprB adhesin, to the cell surface, providing a link between a unique protein translocation system and a motility apparatus in members of the Bacteroidetes phylum. PMID:19966289

14 CFR 121.360 - Ground proximity warning-glide slope deviation alerting system.

Science.gov (United States)

2010-01-01

... deviation alerting system. 121.360 Section 121.360 Aeronautics and Space FEDERAL AVIATION ADMINISTRATION... Equipment Requirements § 121.360 Ground proximity warning-glide slope deviation alerting system. (a) No... system that meets the performance and environmental standards of TSO-C92 (available from the FAA, 800...

Development and validation of duplex, triplex, and pentaplex real-time PCR screening assays for the detection of genetically modified organisms in food and feed.

Science.gov (United States)

Huber, Ingrid; Block, Annette; Sebah, Daniela; Debode, Frédéric; Morisset, Dany; Grohmann, Lutz; Berben, Gilbert; Stebih, Dejan; Milavec, Mojca; Zel, Jana; Busch, Ulrich

2013-10-30

Worldwide, qualitative methods based on PCR are most commonly used as screening tools for genetically modified material in food and feed. However, the increasing number and diversity of genetically modified organisms (GMO) require effective methods for simultaneously detecting several genetic elements marking the presence of transgenic events. Herein we describe the development and validation of a pentaplex, as well as complementary triplex and duplex real-time PCR assays, for the detection of the most common screening elements found in commercialized GMOs: P-35S, T-nos, ctp2-cp4-epsps, bar, and pat. The use of these screening assays allows the coverage of many GMO events globally approved for commercialization. Each multiplex real-time PCR assay shows high specificity and sensitivity with an absolute limit of detection below 20 copies for the targeted sequences. We demonstrate by intra- and interlaboratory tests that the assays are robust as well as cost- and time-effective for GMO screening if applied in routine GMO analysis.

Gliding arc discharge — Application for adhesion improvement of fibre reinforced polyester composites

DEFF Research Database (Denmark)

Kusano, Yukihiro; Teodoru, Steluta; Leipold, Frank

2008-01-01

production, and surface treatment. However, the application for adhesion improvement of structural materials has been rarely reported. In the present work, glass fibre reinforced polyester plates were treated using atmospheric pressure gliding arcs with high speed air flow for adhesion improvement...

Evaluation of canal transportation after preparation with Reciproc single-file systems with or without glide path files.

Science.gov (United States)

Aydin, Ugur; Karataslioglu, Emrah

2017-01-01

Canal transportation is a common sequel caused by rotary instruments. The purpose of the present study is to evaluate the degree of transportation after the use of Reciproc single-file instruments with or without glide path files. Thirty resin blocks with L-shaped canals were divided into three groups ( n = 10). Group 1 - canals were prepared with Reciproc-25 file. Group 2 - glide path file-G1 was used before Reciproc. Group 3 - glide path files-G1 and G2 were used before Reciproc. Pre- and post-instrumentation images were superimposed under microscope, and resin removed from the inner and outer surfaces of the root canal was calculated throughout 10 points. Statistical analysis was performed with Kruskal-Wallis test and post hoc Dunn test. For coronal and middle one-thirds, there was no significant difference among groups ( P > 0.05). For apical section, transportation of Group 1 was significantly higher than other groups ( P files before Reciproc single-file system reduced the degree of apical canal transportation.

Self-interstitial atom clusters as obstacles to glide of 1/3{11-bar 00} edge dislocations in α-zirconium

International Nuclear Information System (INIS)

Voskoboynikov, R.E.; Osetsky, Yu.N.; Bacon, D.J.

2005-01-01

Atomic-scale details of interaction of a 1/3 {11-bar 00} edge dislocation with clusters of self-interstitial atoms (SIAs) in α-zirconium has been studied by computer simulation. Four typical clusters are considered. A triangular cluster of five SIAs lying within a basal plane bisected by the dislocation glide plane is not absorbed by the dislocation but acts as a moderately strong obstacle. A 3-D SIA cluster lying across the glide plane is completely absorbed by the dislocation by creation of super-jogs, and is a weak obstacle. Interaction of the dislocation with glissile SIA loops with perfect Burgers vector inclined at 60 deg. to the dislocation glide plane shows that the process depends on the vector orientation. Defects of the two orientations are strong obstacles, and one, which initially forms a sessile segment on the dislocation line, is particularly so

Gliding arc surface treatment of glass-fiber-reinforced polyester enhanced by ultrasonic irradiation

DEFF Research Database (Denmark)

Kusano, Yukihiro; Norrman, Kion; Drews, Joanna Maria

2011-01-01

. The efficiency of such a plasma treatment at atmospheric pressure can be further improved by ultrasonic irradiation onto the surface during the treatment. In the present work glass fiber reinforced polyester (GFRP) plates are treated using an atmospheric pressure gliding arc with and without ultrasonic...

Dynamics, OH distributions and UV emission of a gliding arc at various flow-rates investigated by optical measurements

DEFF Research Database (Denmark)

Zhu, Jiajian; Sun, Zhiwei; Li, Zhongshan

2014-01-01

-state OH were investigated using planar laser-induced fluorescence. The results show that the shape, height, intensity and thickness of ground-state OH distribution vary significantly with air flow rates. Finally, UV emission of the gliding arc is measured using optical emission spectroscopy......We demonstrate a plasma discharge which is generated between two diverging electrodes and extended into a gliding arc in non-equilibrium condition by an air flow at atmospheric pressure. Effects of the air flow rates on the dynamics, ground-state OH distributions and spectral characterization of UV...

Destruction of tributylphosphate by cold plasma. Use of a gliding arc reactor

International Nuclear Information System (INIS)

Moussa, David

1999-01-01

The nuclear industry uses the Purex process for reprocessing spent nuclear fuel by plutonium and uranium separation. This process uses complexing properties of tributylphosphate. This solvent is aged by the high radioactivity and acidity of the medium and loses its extracting properties. Thus it becomes an highly radioactive liquid organic waste and it must be degraded before its conditioning. We have elaborated a new method for mineralizing TBP by exposure to the plasma produced by a wet air gliding arc. Electric discharges in wet air give rise to very reactive species like excited molecules and radicals. Such species can accelerate oxidation and degradation of organic compounds. The gliding arc discharge is obtained by applying high voltage between two divergent metal electrodes disposed around a blowing nozzle. The arc formed between the electrodes is blown by the air flow with growing in length. Thus a quenched wet air plasma trail is formed and licks an upper layer of TBP while the lower layer is water. Our device can degrade almost 40 percent of the treated TBP. The main degradation product is phosphoric acid for which we have monitored the production kinetics and suggested a model of a surface oxidation process to explain it. Another part of the TBP is converted into a phosphate layer found on the electrodes and phosphorus oxide white smokes present in exhaust fumes. By means of chromatography and spectroscopic analysis we have found the dibutyl-phosphoric acid as the main partial degradation product. The gliding arc device presents several advantages towards other plasma processes which are a low cost and especially for the present task i.e. an easy building and use, operating at atmospheric pressure and moderate temperature, and the possibility to use high powers (several kW for one unit). (author) [fr

Large Gaps in Canopy Reduce Road Crossing by a Gliding Mammal

Directory of Open Access Journals (Sweden)

Rodney van der Ree

2010-12-01

Full Text Available Roads and traffic reduce landscape connectivity and increase rates of mortality for many species of wildlife. Species that glide from tree to tree may be strongly affected by roads and traffic if the size of the gap between trees exceeds their gliding capability. Not only are wide roads likely to reduce crossing rates, but mortality may also be increased if gliders that do cross have poor landing opportunities. The road-crossing behavior of 47 squirrel gliders (Petaurus norfolcensis was investigated in southeast Australia using radio-tracking. The proportion of gliders crossing one or both roadways of a freeway where trees were present or absent from the center median was compared to that at single-lane country roads (control. The proportion of gliders crossing the road at control sites (77% was similar to the proportion that crossed one or both roadways at the freeway with trees in the median (67%, whereas only a single male (6% crossed the freeway where trees were absent from the median. The frequency of crossing for each individual was also similar at control sites and freeway sites with trees in the median. The almost complete lack of crossing at sites where trees were absent from the median was attributed to the wider gap in canopy (50 - 64 m vs. 5 - 13 m at sites with trees in the median. This suggests that traffic volume, up to 5,000 vehicles per day on each roadway, and the other characteristics of the freeway we studied are not in themselves complete deterrents to road crossing by squirrel gliders. This study demonstrates that retaining and facilitating the growth of tall trees in the center median of two-way roads may mitigate the barrier effect of roads on gliders, thus contributing positively to mobility and potentially to connectivity. This information will be essential for the assessment of road impacts on gliding species using population viability models.

Gliding and Quasi-harmonic Tremor Behaviour of Raung Volcano: November 2014 Crisis Period Case Study

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Vico Luthfi Ipmawan

2018-01-01

Full Text Available DOI: 10.17014/ijog.5.1.13-21The seismic activity of Raung Volcano was raised on 11 November 2014. As many as 1709 tremors were recorded followed by continuous tremors appearing in late November 2014. Quasi-harmonic and gliding tremors appeared in a spectrogram on 12 November 2014. The quasi-harmonic tremors refer to tremors that have no fully harmonic form in spectrum. The gliding harmonic tremors refer to harmonic tremors that have frequency jumps with either positive or negative increment. After signal restitution processing, the Maximum Entropy Spectral Analysis (MESA method was applied in Raung recordings resulting the spectrum and the spectrogram of tremors. The quasi-harmonic tremors have the monotonic spectrum in its head and centre segment, and the harmonic one in its tails. There are twenty-four spectrums that show frequency changes between the monotonic and harmonic. The similarity between the fundamental frequency range of the monotonic and harmonic ones suggests that both signals are excited from a common resonator. The alternating of monotonic and harmonic respectively over this period is qualitatively similar with Julian’s synthetic time series about the nonlinear oscillator model. It is suggested that Raung Volcano magma pressure is sizeable to make a chaotic vibration. A pressure increasing in Raung magmatic conduit causes the increasing of P-wave velocity and makes a positive gliding frequency.

Cross-reactivity between methylisothiazolinone, octylisothiazolinone and benzisothiazolinone using a modified local lymph node assay

DEFF Research Database (Denmark)

Schwensen, J. F.; Bonefeld, C. Menne; Zachariae, C.

2017-01-01

, and between MI and BIT. Methods Immune responses to MI, OIT and BIT were studied in vehicle and MI-sensitized female CBA mice by a modified local lymph node assay. The inflammatory response was measured by ear thickness, cell proliferation of CD4+ and CD8+ T cells, and CD19+ B cells in the auricular draining...... lymph nodes. Results MI induced significant, strong, concentration-dependent immune responses in the draining lymph nodes following a sensitization phase of three consecutive days. Groups of MI-sensitized mice were challenged on day 23 with 0·4% MI, 0·7% OIT and 1·9% BIT – concentrations corresponding...

A modified parallel artificial membrane permeability assay for evaluating the bioconcentration of highly hydrophobic chemicals in fish.

Science.gov (United States)

Kwon, Jung-Hwan; Escher, Beate I

2008-03-01

Low cost in vitro tools are needed at the screening stage of assessment of bioaccumulation potential of new and existing chemicals because the number of chemical substances that needs to be tested highly exceeds the capacity of in vivo bioconcentration tests. Thus, the parallel artificial membrane permeability assay (PAMPA) system was modified to predict passive uptake/ elimination rate in fish. To overcome the difficulties associated with low aqueous solubility and high membrane affinity of highly hydrophobic chemicals, we measured the rate of permeation from the donor poly(dimethylsiloxane)(PDMS) disk to the acceptor PDMS disk through aqueous and PDMS membrane boundary layers and term the modified PAMPA system "PDMS-PAMPA". Twenty chemicals were selected for validation of PDMS-PAMPA. The measured permeability is proportional to the passive elimination rate constant in fish and was used to predict the "minimum" in vivo elimination rate constant. The in vivo data were very close to predicted values except for a few polar chemicals and metabolically active chemicals, such as pyrene and benzo[a]pyrene. Thus, PDMS-PAMPA can be an appropriate in vitro system for nonmetabolizable chemicals. Combination with metabolic clearance rates using a battery of metabolic degradation assays would enhance the applicability for metabolizable chemicals.

In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay.

Science.gov (United States)

Hördegen, P; Cabaret, J; Hertzberg, H; Langhans, W; Maurer, V

2006-11-03

Because of the increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against gastrointestinal nematodes. Phytotherapy could be one of the major options to control these pathologies. Extracts or ingredients of six different plant species were tested against exsheathed infective larvae of Haemonchus contortus using a modified methyl-thiazolyl-tetrazolium (MTT) reduction assay. Pyrantel tartrate was used as reference anthelmintic. Bromelain, the enzyme complex of the stem of Ananas comosus (Bromeliaceae), the ethanolic extracts of seeds of Azadirachta indica (Meliaceae), Caesalpinia crista (Caesalpiniaceae) and Vernonia anthelmintica (Asteraceae), and the ethanolic extracts of the whole plant of Fumaria parviflora (Papaveraceae) and of the fruit of Embelia ribes (Myrsinaceae) showed an anthelmintic efficacy of up to 93%, relative to pyrantel tartrate. Based on these results obtained with larval Haemonchus contortus, the modified MTT reduction assay could be a possible method for testing plant products with anthelmintic properties.

Development and Application of Loop-Mediated Isothermal Amplification Assays for Rapid Visual Detection of cry2Ab and cry3A Genes in Genetically-Modified Crops

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Feiwu Li

2014-08-01

Full Text Available The cry2Ab and cry3A genes are two of the most important insect-resistant exogenous genes and had been widely used in genetically-modified crops. To develop more effective alternatives for the quick identification of genetically-modified organisms (GMOs containing these genes, a rapid and visual loop-mediated isothermal amplification (LAMP method to detect the cry2Ab and cry3A genes is described in this study. The LAMP assay can be finished within 60 min at an isothermal condition of 63 °C. The derived LAMP products can be obtained by a real-time turbidimeter via monitoring the white turbidity or directly observed by the naked eye through adding SYBR Green I dye. The specificity of the LAMP assay was determined by analyzing thirteen insect-resistant genetically-modified (GM crop events with different Bt genes. Furthermore, the sensitivity of the LAMP assay was evaluated by diluting the template genomic DNA. Results showed that the limit of detection of the established LAMP assays was approximately five copies of haploid genomic DNA, about five-fold greater than that of conventional PCR assays. All of the results indicated that this established rapid and visual LAMP assay was quick, accurate and cost effective, with high specificity and sensitivity. In addition, this method does not need specific expensive instruments or facilities, which can provide a simpler and quicker approach to detecting the cry2Ab and cry3A genes in GM crops, especially for on-site, large-scale test purposes in the field.

Development and application of loop-mediated isothermal amplification assays for rapid visual detection of cry2Ab and cry3A genes in genetically-modified crops.

Science.gov (United States)

Li, Feiwu; Yan, Wei; Long, Likun; Qi, Xing; Li, Congcong; Zhang, Shihong

2014-08-27

The cry2Ab and cry3A genes are two of the most important insect-resistant exogenous genes and had been widely used in genetically-modified crops. To develop more effective alternatives for the quick identification of genetically-modified organisms (GMOs) containing these genes, a rapid and visual loop-mediated isothermal amplification (LAMP) method to detect the cry2Ab and cry3A genes is described in this study. The LAMP assay can be finished within 60 min at an isothermal condition of 63 °C. The derived LAMP products can be obtained by a real-time turbidimeter via monitoring the white turbidity or directly observed by the naked eye through adding SYBR Green I dye. The specificity of the LAMP assay was determined by analyzing thirteen insect-resistant genetically-modified (GM) crop events with different Bt genes. Furthermore, the sensitivity of the LAMP assay was evaluated by diluting the template genomic DNA. Results showed that the limit of detection of the established LAMP assays was approximately five copies of haploid genomic DNA, about five-fold greater than that of conventional PCR assays. All of the results indicated that this established rapid and visual LAMP assay was quick, accurate and cost effective, with high specificity and sensitivity. In addition, this method does not need specific expensive instruments or facilities, which can provide a simpler and quicker approach to detecting the cry2Ab and cry3A genes in GM crops, especially for on-site, large-scale test purposes in the field.

Immunosuppressive potential of bardoxolone methyl using a modified murine local lymph node assay (LLNA).

Science.gov (United States)

Kitsukawa, Mika; Tsuchiyama, Hiromi; Maeda, Akihisa; Oshida, Keiyu; Miyamoto, Yohei

2014-08-01

2-Cyano-3, 12-dioxooleana-1, 9-dien-28-oic acid methyl ester (CDDO-Me; bardoxolone methyl) is one of the synthetic oleanane triterpenoids (SOs). It is known that it is the strongest Nrf2/ARE signaling inducer of SOs and slightly inhibits immune response. Little was known about the immunomodulatory action of CDDO-Me in vivo. We assessed its immunosuppressive potential by using the modified mouse lymph node assay (LLNA) including immunosuppression-related gene expression analysis. In the modified LLNA, CDDO-Me showed a significant decrease in lymph node weight and changes in expressions of the immunosuppression-related genes, Zfp459 and Fmo2. It has been already reported that a decrease in lymph node weight was induced by several types of immunosuppressive chemicals such as calcineurin inhibitors, antimetabolites, steroids, and alkylators. In addition, changes in Zfp459 and Fmo2 expression was reported in response after only treatment of antimetabolites. From these results, CDDO-Me is considered to have an immunosuppressive action and similar mechanism to antimetabolites.

Endogenous Reference Genes and Their Quantitative Real-Time PCR Assays for Genetically Modified Bread Wheat (Triticum aestivum L.) Detection.

Science.gov (United States)

Yang, Litao; Quan, Sheng; Zhang, Dabing

2017-01-01

Endogenous reference genes (ERG) and their derivate analytical methods are standard requirements for analysis of genetically modified organisms (GMOs). Development and validation of suitable ERGs is the primary step for establishing assays that monitoring the genetically modified (GM) contents in food/feed samples. Herein, we give a review of the ERGs currently used for GM wheat analysis, such as ACC1, PKABA1, ALMT1, and Waxy-D1, as well as their performances in GM wheat analysis. Also, we discussed one model for developing and validating one ideal RG for one plant species based on our previous research work.

The Effects of One-Dimensional Glide on the Reaction Kinetics of Interstitial Clusters

International Nuclear Information System (INIS)

Heinisch, Howard L.; Singh, B N.; Golubov, S I.

2000-01-01

Collision cascades in metals produce small interstitial clusters and perfect dislocation loops that glide in thermally activated one-dimensional (1D) random walks. These gliding defects can change their Burgers vectors by thermal activation or by interactions with other defects. Their migration is therefore''mixed 1D/3D migration'' along a 3D path consisting of 1D segments. The defect reaction kinetics under mixed 1D/3D diffusion are different from pure 1D diffusion and pure 3D diffusion, both of which can be formulated within analytical rate theory models of microstructure evolution under irradiation. Atomic-scale kinetic Monte Carlo (kMC) defect migration simulations are used to investigate the effects of mixed 1D/3D migration on defect reaction kinetics as a guide for implementing mixed 1D/3D migration into the analytical rate theory. The functional dependence of the sink strength on the sixe and concentration of sinks under mixed 1D/3D migration is shown to lie between that for pure 1D and pure 3D migration and varies with L, the average distance between direction changes of the gliding defects. It is shown that the sink strength in simulations for spherical sinks of radius R under mixed 1D/3D migration for values of L greater than R can be approximated by an expression that varies directly as R2. For small L, the form of the transition from mixed 1D/3D to pure 3D diffusion as L decreases is demonstrated in the simulations, the results of which can be used in the future development of an analytical expression describing this transition region

Standardization and performance evaluation of "modified" and "ultrasensitive" versions of the Abbott RealTime HIV-1 assay, adapted to quantify minimal residual viremia.

Science.gov (United States)

Amendola, Alessandra; Bloisi, Maria; Marsella, Patrizia; Sabatini, Rosella; Bibbò, Angela; Angeletti, Claudio; Capobianchi, Maria Rosaria

2011-09-01

Numerous studies investigating clinical significance of HIV-1 minimal residual viremia (MRV) suggest potential utility of assays more sensitive than those routinely used to monitor viral suppression. However currently available methods, based on different technologies, show great variation in detection limit and input plasma volume, and generally suffer from lack of standardization. In order to establish new tools suitable for routine quantification of minimal residual viremia in patients under virological suppression, some modifications were introduced into standard procedure of the Abbott RealTime HIV-1 assay leading to a "modified" and an "ultrasensitive" protocols. The following modifications were introduced: calibration curve extended towards low HIV-1 RNA concentration; 4 fold increased sample volume by concentrating starting material; reduced volume of internal control; adoption of "open-mode" software for quantification. Analytical performances were evaluated using the HIV-1 RNA Working Reagent 1 for NAT assays (NIBSC). Both tests were applied to clinical samples from virologically suppressed patients. The "modified" and the "ultrasensitive" configurations of the assay reached a limit of detection of 18.8 (95% CI: 11.1-51.0 cp/mL) and 4.8 cp/mL (95% CI: 2.6-9.1 cp/mL), respectively, with high precision and accuracy. In clinical samples from virologically suppressed patients, "modified" and "ultrasensitive" protocols allowed to detect and quantify HIV RNA in 12.7% and 46.6%, respectively, of samples resulted "not-detectable", and in 70.0% and 69.5%, respectively, of samples "detected laboratories for measuring MRV. Copyright © 2011 Elsevier B.V. All rights reserved.

Translational, rotational and vibrational temperatures of a gliding arc discharge at atmospheric pressure air

DEFF Research Database (Denmark)

Zhu, Jiajian; Gao, Jinlong; Ehn, Andreas

2014-01-01

and vibrational temperatures of a gliding arc generated at atmospheric pressure air are investigated. Translational temperatures (about 1100 K) were measured by laser-induced Rayleigh scattering, and two-dimensional temperature imaging was performed. Rotational and vibrational temperatures (about 3600 K and 6700...

Relationship between the electronic structure and the glide in the hexagonal close packed metals

International Nuclear Information System (INIS)

Legrand, B.; Le Hazif, R.

1983-06-01

In all hexagonal close-packed metals (HCP), deformation is performed by slip on a mean glide system (MGS) and on several secondary systems. There are no reliable predictions of the MGS choice. In this paper is shown the role played by the electronic structure on the choice of glide system in HCP metals. MGS is basal for all normal metals and is a function of the electron number in HCP transition metals. The different SFE's were calculated using appropriate total energy models, for different metals. Thus pseudopotentials were used (or empirical pair potentials) for normal metals, and a tight-binding model for transition metals. The most important results are the following: prismatic SFE (PSFE) is smaller than basal SFE (BSFE) for Y, Ti, Zr, Hf, Ru and Os; BSFE is smaller than PSFE for Co and all normal metals; BSFE and PSFe and about the same for RE and Tc

The barrier to misfit dislocation glide in continuous, strained, epitaxial layers on patterned substrates

International Nuclear Information System (INIS)

Watson, G.P.; Ast, D.G.; Anderson, T.J.; Pathangey, B.

1993-01-01

In a previous report [G. P. Watson, D. G. Ast, T. J. Anderson, and Y. Hayakawa, Appl. Phys. Lett. 58, 2517 (1991)] we demonstrated that the motion of misfit dislocations in InGaAs, grown by organometallic vapor phase epitaxy on patterned GaAs substrates, can be impeded even if the strained epitaxial layer is continuous. Trenches etched into GaAs before growth are known to act as a barrier to misfit dislocation propagation [E. A. Fitzgerald, G. P. Watson, R. E. Proano, D. G. Ast, P. D. Kirchner, G. D. Pettit, and J. M. Woodall, J. Appl. Phys. 65, 2220 (1989)] when those trenches create discontinuities in the epitaxial layers; but even shallow trenches, with continuous strained layers following the surface features, can act as barriers. By considering the strain energy required to change the length of the dislocation glide segments that stretch from the interface to the free surface, a simple model is developed that explains the major features of the unique blocking action observed at the trench edges. The trench wall angle is found to be an important parameter in determining whether or not a trench will block dislocation glide. The predicted blocking angles are consistent with observations made on continuous 300 and 600 nm thick In 0.04 Ga 0.96 As films on patterned GaAs. Based on the model, a structure is proposed that may be used as a filter to yield misfit dislocations with identical Burgers vectors or dislocations which slip in only one glide plane

CR TKA UHMWPE Wear Tested after Artificial Aging of the Vitamin E Treated Gliding Component by Simulating Daily Patient Activities

Directory of Open Access Journals (Sweden)

Jens Schwiesau

2014-01-01

Full Text Available The wear behaviour of total knee arthroplasty (TKA is dominated by two wear mechanisms: the abrasive wear and the delamination of the gliding components, where the second is strongly linked to aging processes and stress concentration in the material. The addition of vitamin E to the bulk material is a potential way to reduce the aging processes. This study evaluates the wear behaviour and delamination susceptibility of the gliding components of a vitamin E blended, ultra-high molecular weight polyethylene (UHMWPE cruciate retaining (CR total knee arthroplasty. Daily activities such as level walking, ascending and descending stairs, bending of the knee, and sitting and rising from a chair were simulated with a data set received from an instrumented knee prosthesis. After 5 million test cycles no structural failure of the gliding components was observed. The wear rate was with 5.62±0.53 mg/million cycles falling within the limit of previous reports for established wear test methods.

CR TKA UHMWPE wear tested after artificial aging of the vitamin E treated gliding component by simulating daily patient activities.

Science.gov (United States)

Schwiesau, Jens; Fritz, Bernhard; Kutzner, Ines; Bergmann, Georg; Grupp, Thomas M

2014-01-01

The wear behaviour of total knee arthroplasty (TKA) is dominated by two wear mechanisms: the abrasive wear and the delamination of the gliding components, where the second is strongly linked to aging processes and stress concentration in the material. The addition of vitamin E to the bulk material is a potential way to reduce the aging processes. This study evaluates the wear behaviour and delamination susceptibility of the gliding components of a vitamin E blended, ultra-high molecular weight polyethylene (UHMWPE) cruciate retaining (CR) total knee arthroplasty. Daily activities such as level walking, ascending and descending stairs, bending of the knee, and sitting and rising from a chair were simulated with a data set received from an instrumented knee prosthesis. After 5 million test cycles no structural failure of the gliding components was observed. The wear rate was with 5.62 ± 0.53 mg/million cycles falling within the limit of previous reports for established wear test methods.

Development of an event-specific hydrolysis probe quantitative real-time polymerase chain reaction assay for Embrapa 5.1 genetically modified common bean (Phaseolus vulgaris).

Science.gov (United States)

Treml, Diana; Venturelli, Gustavo L; Brod, Fábio C A; Faria, Josias C; Arisi, Ana C M

2014-12-10

A genetically modified (GM) common bean event, namely Embrapa 5.1, resistant to the bean golden mosaic virus (BGMV), was approved for commercialization in Brazil. Brazilian regulation for genetically modified organism (GMO) labeling requires that any food containing more than 1% GMO be labeled. The event-specific polymerase chain reaction (PCR) method has been the primary trend for GMO identification and quantitation because of its high specificity based on the flanking sequence. This work reports the development of an event-specific assay, named FGM, for Embrapa 5.1 detection and quantitation by use of SYBR Green or hydrolysis probe. The FGM assay specificity was tested for Embrapa 2.3 event (a noncommercial GM common bean also resistant to BGMV), 46 non-GM common bean varieties, and other crop species including maize, GM maize, soybean, and GM soybean. The FGM assay showed high specificity to detect the Embrapa 5.1 event. Standard curves for the FGM assay presented a mean efficiency of 95% and a limit of detection (LOD) of 100 genome copies in the presence of background DNA. The primers and probe developed are suitable for the detection and quantitation of Embrapa 5.1.

Cyclic fatigue resistance of R-Pilot, WaveOne Gold Glider, and ProGlider glide path instruments.

Science.gov (United States)

Keskin, Cangül; İnan, Uğur; Demiral, Murat; Keleş, Ali

2018-02-17

The aim of the present study was to compare the cyclic fatigue resistance of R-Pilot (VDW; Munich, Germany) with ProGlider (Denstply Sirona; Ballaigues, Switzerland) and WaveOne Gold Glider (Denstply Sirona; Ballaigues, Switzerland) glide path instruments. R-Pilot, ProGlider, and WaveOne Gold Glider instruments were collected (n = 15) and tested in a dynamic cyclic fatigue test device, which has an artificial canal with 60° angle of curvature and a 5-mm radius of curvature. All instruments were operated until fracture occurred, and both time to fracture (TF) and the lengths of the fractured fragments were recorded. Mean and standard deviations of TF and fragment length were calculated for each reciprocating system. TF data and fractured fragment length data were subjected to one-way ANOVA and post-hoc Tukey tests (P  0.05). Weibull analysis revealed that WaveOne Gold Glider showed the highest predicted TF value for 99% survival rate, which was followed by R-Pilot and ProGlider. Regarding the length of the fractured tips, there were no significant differences among the instruments (P > 0.05). The reciprocating WaveOne Gold Glider and R-Pilot instruments had significantly higher cyclic fatigue resistance than rotary ProGlider instruments. This study reported that novel reciprocating glide path instruments exhibited higher cyclic fatigue resistance than rotating glide path instrument.

Gravity effects on a gliding arc in four noble gases: from normal to hypergravity

NARCIS (Netherlands)

Potocnakova, L.; Sperka, J.; Zikan, P.; van Loon, J.J.W.A.; Beckers, J.; Kudrle, V.

2015-01-01

A gliding arc in four noble gases (He, Ne, Ar, Kr) has been studied under previously unexplored conditions of varying artificial gravity, from normal 1 g gravity up to 18 g hypergravity. Significant differences, mainly the visual thickness of the plasma channel, its maximum elongation and general

Glide path preparation in S-shaped canals with rotary pathfinding nickel-titanium instruments.

Science.gov (United States)

Ajuz, Natasha C C; Armada, Luciana; Gonçalves, Lucio S; Debelian, Gilberto; Siqueira, José F

2013-04-01

This study compared the incidence of deviation along S-shaped (double-curved) canals after glide path preparation with 2 nickel-titanium (NiTi) rotary pathfinding instruments and hand K-files. S-shaped canals from 60 training blocks were filled with ink, and preinstrumentation images were obtained by using a stereomicroscope. Glide path preparation was performed by an endodontist who used hand stainless steel K-files (up to size 20), rotary NiTi PathFile instruments (up to size 19), or rotary NiTi Scout RaCe instruments (up to size 20). Postinstrumentation images were taken by using exactly the same conditions as for the preinstrumentation images, and both pictures were superimposed. Differences along the S-shaped canal for the mesial and distal aspects were measured to evaluate the occurrence of deviation. Intragroup analysis showed that all instruments promoted some deviation in virtually all levels. Overall, regardless of the group, deviations were observed in the mesial wall at the canal terminus and at levels 4, 5, 6 and 7 mm and in the distal wall at levels 1, 2, and 3 mm. These levels corresponded to the inner walls of each curvature. Both rotary NiTi instruments performed significantly better than hand K-files at all levels (P instruments showed significantly better results than PathFiles at levels 0, 2, 3, 5, and 6 mm (P rotary NiTi instruments are suitable for adequate glide path preparation because they promoted less deviation from the original canal anatomy when compared with hand-operated instruments. Of the 2 rotary pathfinding instruments, Scout RaCe showed an overall significantly better performance. Copyright © 2013 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Apical Transportation, Centering Ability, and Cleaning Effectiveness of Reciprocating Single-file System Associated with Different Glide Path Techniques.

Science.gov (United States)

de Carvalho, Guilherme Moreira; Sponchiado Junior, Emílio Carlos; Garrido, Angela Delfina Bittencourt; Lia, Raphael Carlos Comelli; Garcia, Lucas da Fonseca Roberti; Marques, André Augusto Franco

2015-12-01

The aim of this study was to evaluate the apical transportation, the centering ability, and the cleaning effectiveness of a reciprocating single-file system associated to different glide path techniques. The mesial root canals of 52 mandibular molars were randomly distributed into 4 groups (n = 13) according to the different glide path techniques used before biomechanical preparation with Reciproc System (RS): KF/RS (sizes 10 and 15 K-files), NGP/RS (no glide path, only reciprocating system), PF/RS (sizes 13, 16, and 19 PathFile instruments), and NP (no preparation). Cone-beam computed tomography analysis was performed before and after instrumentation for apical third images acquisition. Apical transportation and its direction were evaluated by using the formula D = (X1 - X2) - (Y1 - Y2), and the centering ability was analyzed by the formula CC = (X1 - X2/Y1 - Y2 or Y1 - Y2/X1 - X2). The samples were submitted to histologic processing and analyzed under a digital microscope for debris quantification. The values were statistically analyzed (Kruskal-Wallis, the Dunn multiple comparisons test, P .05). Groups had a tendency toward transportation in the mesial direction. No technique had perfect centering ability (=1.0), with no significant difference among them. KF/RS had larger amount of debris, with statistically significant difference in comparison with NGP/RS (P > .05). The different glide path techniques promoted minimal apical transportation, and the reciprocating single-file system tested remained relatively centralized within the root canal. Also, the different techniques interfered in the cleaning effectiveness of the reciprocating system. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Comparison of Friedewald Formula and Modified Friedewald Formula with Direct Homogeneous Assay for Low Density Lipoprotein Cholesterol Estimation

International Nuclear Information System (INIS)

Anwar, M.; Khan, D. A.; Khan, F. A.

2014-01-01

Objective: To compare the Friedewald and modified Friedewald formulae with direct homogeneous assay for serum lowdensity lipoprotein cholesterol (LDL-C) levels estimation. Study Design: Cross-sectional study. Place and Duration of Study: Armed Forces Institute of Pathology, Rawalpindi, from June to December 2011. Methodology: Healthy subjects of either gender, from Rawalpindi, aged 18-75 years were included by consecutive sampling. Patients with diabetes mellitus, chronic liver disease, chronic kidney disease, those taking lipid lowering drugs and samples with triglyceride (TG) > 4.52 mmol/l were excluded from the study. Total cholesterol, high-density lipoprotein cholesterol, TG and LDL-C were measured on Hitachi 912 chemistry analyzer (Roche). LDL-C levels were also calculated by Friedewald formula (FF) and Vujovic modified formula (VMF). Paired sample t-test and scatter plots were used for statistical analysis. Results: Although both calculated methods showed good correlation with direct assay (r > 0.93) in 300 subjects, but the difference was statistically significant. The ffLDL-C were 0.12 +- 31 mmol/l (p < 0.001) lower and vmfLDL-C were 0.11 +- 26 mmol/l (p < 0.001) higher than dLDL-C. The difference was not significant between ffLDL-C and dLDL-C at TG levels < 1.70 mmol/l (p = 0.58) and between vmfLDL-C and dLDL-C at TG levels 2.26 - 4.52 mmol/l (p = 0.38). At all other TG levels, the difference between LDL-C calculated by both formulas and dLDL-C was statistically significant (p < 0.001). As compared to direct assay, 11% and 14% subjects were classified in wrong National Cholesterol Education Programm cardiac risk categories by FF and VMF respectively. Conclusion: LDL-C should be measured by direct homogeneous assay in routine clinical laboratories, as the calculated methods did not have a uniform performance for LDL-C estimation at different TG levels. (author)

A Prospective Observational Study of Technical Difficulty With GlideScope-Guided Tracheal Intubation in Children.

Science.gov (United States)

Zhang, Bin; Gurnaney, Harshad G; Stricker, Paul A; Galvez, Jorge A; Isserman, Rebecca S; Fiadjoe, John E

2018-05-09

The GlideScope Cobalt is one of the most commonly used videolaryngoscopes in pediatric anesthesia. Although visualization of the airway may be superior to direct laryngoscopy, users need to learn a new indirect way to insert the tracheal tube. Learning this indirect approach requires focused practice and instruction. Identifying the specific points during tube placement, during which clinicians struggle, would help with targeted education. We conducted this prospective observational study to determine the incidence and location of technical difficulties using the GlideScope, the success rates of various corrective maneuvers used, and the impact of technical difficulty on success rate. We conducted this observational study at our quaternary pediatric hospital between February 2014 and August 2014. We observed 200 GlideScope-guided intubations and documented key intubation-related outcomes. Inclusion criteria for patients were the number of advancement maneuvers required to intubate the trachea, the location where technical difficulty occurred, the types of maneuvers used to address difficulties, and the tracheal intubation success rate. We used a bias-corrected bootstrapping method with 300 replicates to determine the 95% confidence interval (CI) around the rate of difficulty with an intubation attempt. After excluding attempts by inexperienced clinicians, there were 225 attempts in 187 patients, 58% (131 of 225; bootstrap CI, 51.6%-64.6%]) of the attempts had technical difficulties. Technical difficulty was most likely to occur when inserting the tracheal tube between the plane of the arytenoid cartilages to just beyond the vocal cords: "zone 3." Clockwise rotation of the tube was the most common successful corrective maneuver in zone 3. The overall tracheal intubation success rate was 98% (CI, 95%-99%); however, the first attempt success rate was only 80% (CI, 74%-86%). Patients with technical difficulty had more attempts (median [interquartile range], 2 [1

Anteroposterior glide versus rotating platform low contact stress (LCS knee arthroplasty: a randomised controlled trial

Directory of Open Access Journals (Sweden)

Wynn-Jones Charles

2007-08-01

Full Text Available Abstract Background Fifty thousand knee replacements are performed annually in the UK at an estimated cost of £150 million. Post-operative improvement depends on a number of factors including implant design and patient associated factors. To our knowledge there are no published study's comparing the results of AP glide and rotating platform designs of LCS knee arthroplasty. Therefore we feel that a study is required to investigate and compare the effects of two types of LCS total knee arthroplasty on joint proprioception and range of motion. Methods/Design Patients will be randomised to receive either a LCS AP glide or Rotating platform prosthesis. Clinical scores (Oxford knee score, American knee society score, EuroQol, range of motion and proprioception will be assessed prior to and at 3,6, 12 and 24 months after the operation. Proprioception will be assessed in terms of absolute error angle (mean difference between the target angle and the response angle. Knee angles will be measured in degrees using an electromagnetic tracking device, Polhemus 3Space Fastrak that detects positions of sensors placed on the test limb. Student's t-test will be used to compare the mean of two groups. Discussion Evidence is lacking concerning the best prosthesis to use for patients undergoing total knee replacement. This pragmatic randomised trial will test the null hypothesis that anteroposterior glide LCS knee arthroplasty does not result in better post operative knee motion and proprioception as compared to rotating platform LCS knee. Trial Registration ISRCTN52943804

Arc dynamics of a pulsed DC nitrogen rotating gliding arc discharge

Science.gov (United States)

Zhu, Fengsen; Zhang, Hao; Li, Xiaodong; Wu, Angjian; Yan, Jianhua; Ni, Mingjiang; Tu, Xin

2018-03-01

In this study, a novel pulsed direct current (DC) rotating gliding arc (RGA) plasma reactor co-driven by an external magnetic field and a tangential gas flow has been developed. The dynamic characteristics of the rotating gliding arc have been investigated by means of numerical simulation and experiment. The simulation results show that a highly turbulent vortex flow can be generated at the bottom of the RGA reactor to accelerate the arc rotation after arc ignition, whereas the magnitude of gas velocity declined significantly along the axial direction of the RGA reactor. The calculated arc rotation frequency (14.4 Hz) is reasonably close to the experimental result (18.5 Hz) at a gas flow rate of 10 l min-1. In the presence of an external magnet, the arc rotation frequency is around five times higher than that of the RGA reactor without using a magnet, which suggests that the external magnetic field plays a dominant role in the maintenance of the arc rotation in the upper zone of the RGA reactor. In addition, when the magnet is placed outside the reactor reversely to form a reverse external magnetic field, the arc can be stabilized at a fixed position in the inner wall of the outer electrode at a critical gas flow rate of 16 l min-1.

Leaping shampoo glides on a lubricating air layer

Science.gov (United States)

Lee, S.; Li, E. Q.; Marston, J. O.; Bonito, A.; Thoroddsen, S. T.

2013-06-01

When a stream of shampoo is fed onto a pool in one's hand, a jet can leap sideways or rebound from the liquid surface in an intriguing phenomenon known as the Kaye effect. Earlier studies have debated whether non-Newtonian effects are the underlying cause of this phenomenon, making the jet glide on top of a shear-thinning liquid layer, or whether an entrained air layer is responsible. Herein we show unambiguously that the jet slides on a lubricating air layer. We identify this layer by looking through the pool liquid and observing its rupture into fine bubbles. The resulting microbubble sizes suggest this air layer is of submicron thickness. This thickness estimate is also supported by the tangential deceleration of the jet during the rebounding.

Leaping shampoo glides on a lubricating air layer

KAUST Repository

Lee, S.

2013-06-10

When a stream of shampoo is fed onto a pool in one\\'s hand, a jet can leap sideways or rebound from the liquid surface in an intriguing phenomenon known as the Kaye effect. Earlier studies have debated whether non-Newtonian effects are the underlying cause of this phenomenon, making the jet glide on top of a shear-thinning liquid layer, or whether an entrained air layer is responsible. Herein we show unambiguously that the jet slides on a lubricating air layer. We identify this layer by looking through the pool liquid and observing its rupture into fine bubbles. The resulting microbubble sizes suggest this air layer is of submicron thickness. This thickness estimate is also supported by the tangential deceleration of the jet during the rebounding.

Leaping shampoo glides on a lubricating air layer

KAUST Repository

Lee, S.; Li, Erqiang; Marston, J. O.; Bonito, A.; Thoroddsen, Sigurdur T

2013-01-01

When a stream of shampoo is fed onto a pool in one's hand, a jet can leap sideways or rebound from the liquid surface in an intriguing phenomenon known as the Kaye effect. Earlier studies have debated whether non-Newtonian effects are the underlying cause of this phenomenon, making the jet glide on top of a shear-thinning liquid layer, or whether an entrained air layer is responsible. Herein we show unambiguously that the jet slides on a lubricating air layer. We identify this layer by looking through the pool liquid and observing its rupture into fine bubbles. The resulting microbubble sizes suggest this air layer is of submicron thickness. This thickness estimate is also supported by the tangential deceleration of the jet during the rebounding.

Cyclic fatigue resistances of several nickel-titanium glide path rotary and reciprocating instruments at body temperature.

Science.gov (United States)

Yılmaz, K; Uslu, G; Gündoğar, M; Özyürek, T; Grande, N M; Plotino, G

2018-01-31

To compare the cyclic fatigue resistance of the One G, ProGlider, HyFlex EDM and R-Pilot glide path NiTi files at body temperature. Twenty One G (size 14, .03 taper), 20 ProGlider (size 16, .02 taper), 20 HyFlex EDM (size 10, .05 taper) and 20 R-Pilot (size 12.5, .04 taper) instruments were operated in rotation at 300 rpm (One G, ProGlider and HyFlex) or in reciprocation (R-Pilot) at 35 °C in artificial canals that were manufactured by reproducing the size and taper of the instrument until fracture occurred. The time to fracture was recorded in seconds using a digital chronometer, and the length of the fractured fragments was registered. Mean data were analysed statistically using the Kruskal-Wallis test and post hoc Tukey tests via SPSS 21.0 software. The statistical significance level was set at 5%. The cyclic fatigue resistance of the R-Pilot files was significantly greater than the other instruments, and the One G was significantly lower (P EDM and the ProGlider (P > 0.05). No significant difference (P > 0.05) was evident in the mean length of the fractured fragments of the various instruments. The cyclic fatigue resistance of the R-Pilot reciprocating glide path file was significantly greater than that of the rotary HyFlex EDM, ProGlider and One G glide path files. © 2018 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Spatiotemporally resolved characteristics of a gliding arc discharge in a turbulent air flow at atmospheric pressure

DEFF Research Database (Denmark)

Zhu, Jiajian; Gao, Jinlong; Ehn, Andreas

2017-01-01

A gliding arc discharge was generated in a turbulent air flow at atmospheric pressure driven by a 35 kHz alternating current (AC) electric power. The spatiotemporally resolved characteristics of the gliding arc discharge, including glow-type discharges, spark-type discharges, short-cutting events...... and transitions among the different types of discharges, were investigated using simultaneously optical and electrical diagnostics. The glow-type discharge shows sinusoidal-like voltage and current waveforms with a peak current of hundreds of milliamperes. The frequency of the emission intensity variation...... of the glow-type discharge is the same as that of the electronic power dissipated in the plasma column. The glow-type discharge can transfer into a spark discharge characterized by a sharp peak current of several amperes and a sudden increase of the brightness in the plasma column. Transitions can also...

Effect of Glide Path Creating on Cyclic Fatigue Resistance of Reciproc and Reciproc Blue Nickel-titanium Files: A Laboratory Study.

Science.gov (United States)

Özyürek, Taha; Uslu, Gülşah; Yılmaz, Koray; Gündoğar, Mustafa

2018-06-01

The purpose of this article was to compare the cyclic fatigue resistance of Reciproc and Reciproc Blue files (VDW GmbH, Munich, Germany) that were used to prepare root canals of mandibular molar teeth with or without a glide path. Sixty Reciproc R25 and 60 Reciproc Blue R25 files were used. The Reciproc and Reciproc Blue groups were divided into 3 subgroups (ie, as received condition, used without a glide path, and used with a glide path). All the instruments were rotated in a stainless steel artificial canal with an inner diameter of 1.5 mm, a 60° angle of curvature, and a radius of curvature of 5 mm until fracture occurred. The number of cycle to fracture was calculated, and the length of the fractured segments was measured. The Kruskal-Wallis test was performed to statistically analyze the data using SPSS 21.0 software (IBM Corp, Armonk, NY) at a 5% significance level. The cyclic fatigue resistance of as received condition Reciproc Blue files was found to be higher than as received condition Reciproc files (P  .05). There was no statistically significant difference in the mean length of the fractured fragments of the instruments (P > .05). Within the limitations of this in vitro study, it was concluded that creating a glide path using ProGlider files had no effect on the cyclic fatigue resistance of RPC and RPC Blue files. Copyright © 2018 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Deformation behavior of dragonfly-inspired nodus structured wing in gliding flight through experimental visualization approach.

Science.gov (United States)

Zhang, Sheng; Sunami, Yuta; Hashimoto, Hiromu

2018-04-10

Dragonfly has excellent flight performance and maneuverability due to the complex vein structure of wing. In this research, nodus as an important structural element of the dragonfly wing is investigated through an experimental visualization approach. Three vein structures were fabricated as, open-nodus structure, closed-nodus structure (with a flex-limiter) and rigid wing. The samples were conducted in a wind tunnel with a high speed camera to visualize the deformation of wing structure in order to study the function of nodus structured wing in gliding flight. According to the experimental results, nodus has a great influence on the flexibility of the wing structure. Moreover, the closed-nodus wing (with a flex-limiter) enables the vein structure to be flexible without losing the strength and rigidity of the joint. These findings enhance the knowledge of insect-inspired nodus structured wing and facilitate the application of Micro Air Vehicle (MAV) in gliding flight.

Effect of repetitive pecking at working length for glide path preparation using G-file

Directory of Open Access Journals (Sweden)

Jung-Hong Ha

2015-05-01

Full Text Available Objectives Glide path preparation is recommended to reduce torsional failure of nickel-titanium (NiTi rotary instruments and to prevent root canal transportation. This study evaluated whether the repetitive insertions of G-files to the working length maintain the apical size as well as provide sufficient lumen as a glide path for subsequent instrumentation. Materials and Methods The G-file system (Micro-Mega composed of G1 and G2 files for glide path preparation was used with the J-shaped, simulated resin canals. After inserting a G1 file twice, a G2 file was inserted to the working length 1, 4, 7, or 10 times for four each experimental group, respectively (n = 10. Then the canals were cleaned by copious irrigation, and lubricated with a separating gel medium. Canal replicas were made using silicone impression material, and the diameter of the replicas was measured at working length (D0 and 1 mm level (D1 under a scanning electron microscope. Data was analysed by one-way ANOVA and post-hoc tests (p = 0.05. Results The diameter at D0 level did not show any significant difference between the 1, 2, 4, and 10 times of repetitive pecking insertions of G2 files at working length. However, 10 times of pecking motion with G2 file resulted in significantly larger canal diameter at D1 (p < 0.05. Conclusions Under the limitations of this study, the repetitive insertion of a G2 file up to 10 times at working length created an adequate lumen for subsequent apical shaping with other rotary files bigger than International Organization for Standardization (ISO size 20, without apical transportation at D0 level.

In vitro screening of six anthelmintic plant products against larval Haemonchus contortus with a modified methyl-thiazolyl-tetrazolium reduction assay

OpenAIRE

Hördegen, P.; Cabaret, J.; Hertzberg, H.; Langhans, W.; Maurer, V.

2006-01-01

Because of the increasing anthelmintic resistance and the impact of conventional anthelmintics on the environment, it is important to look for alternative strategies against gastrointestinal nematodes. Phytotherapy could be one of the major options to control these pathologies. Extracts or ingredients of six different plant species were tested against exsheathed infective larvae of Haemonchus contortus using a modified methyl-thiazolyltetrazolium (MTT) reduction assay. Pyrantel tartrate was u...

Preservation of root canal anatomy using self-adjusting file instrumentation with glide path prepared by 20/0.02 hand files versus 20/0.04 rotary files

Science.gov (United States)

Jain, Niharika; Pawar, Ajinkya M.; Ukey, Piyush D.; Jain, Prashant K.; Thakur, Bhagyashree; Gupta, Abhishek

2017-01-01

Objectives: To compare the relative axis modification and canal concentricity after glide path preparation with 20/0.02 hand K-file (NITIFLEX®) and 20/0.04 rotary file (HyFlex™ CM) with subsequent instrumentation with 1.5 mm self-adjusting file (SAF). Materials and Methods: One hundred and twenty ISO 15, 0.02 taper, Endo Training Blocks (Dentsply Maillefer, Ballaigues, Switzerland) were acquired and randomly divided into following two groups (n = 60): group 1, establishing glide path till 20/0.02 hand K-file (NITIFLEX®) followed by instrumentation with 1.5 mm SAF; and Group 2, establishing glide path till 20/0.04 rotary file (HyFlex™ CM) followed by instrumentation with 1.5 mm SAF. Pre- and post-instrumentation digital images were processed with MATLAB R 2013 software to identify the central axis, and then superimposed using digital imaging software (Picasa 3.0 software, Google Inc., California, USA) taking five landmarks as reference points. Student's t-test for pairwise comparisons was applied with the level of significance set at 0.05. Results: Training blocks instrumented with 20/0.04 rotary file and SAF were associated less deviation in canal axis (at all the five marked points), representing better canal concentricity compared to those, in which glide path was established by 20/0.02 hand K-files followed by SAF instrumentation. Conclusion: Canal geometry is better maintained after SAF instrumentation with a prior glide path established with 20/0.04 rotary file. PMID:28855752

Development of a plutonium solution-assay instrument with isotopic capability

International Nuclear Information System (INIS)

Hsue, S.T.; Marks, T.

1992-01-01

A new generation of solution-assay instrument has been developed to satisfy all the assay requirements of an aqueous plutonium-recovery operation. The assay is based on a transmission-corrected passive assay technique. We have demonstrated that the system can cover a concentration range of 0.5--300 g/ell with simultaneous isotopic determination. The system can be used to assay input and eluate streams of the recovery operation. The system can be modified to measure low-concentration effluent solutions from the recovery operation covering 0.01--40 g/ell. The same system has also been modified to assay plutonium solutions enriched in 242 Pu. 6 refs

Genotoxicity of unmodified and organo-modified montmorillonite

DEFF Research Database (Denmark)

Sharma, Anoop Kumar; Schmidt, Bjørn; Frandsen, Henrik Lauritz

2010-01-01

absent in the filtered samples, which was independently confirmed by dynamic light-scattering measurements. Detection and identification of free quaternary ammonium modifier in the filtered sample was carried out by HPLC-Q-TOF/MS and revealed a total concentration of a mixture of quaternary ammonium...... assay, none of the clays produced ROS in a cell-free test system (the DCFH-DA assay). Inductively coupled plasma mass-spectrometry (ICP-MS) was used to detect clay particles in the filtered samples using aluminium as a tracer element characteristic to clay. The results indicated that clay particles were...... analogues of 1.57 mu g/ml. These findings suggest that the genotoxicity of organo-modified montmorillonite was caused by the organo-modifier. The detected organo-modifier mixture was synthesized and comet-assay results showed that the genotoxic potency of this synthesized organo-modifier was in the same...

Sustained diffusive alternating current gliding arc discharge in atmospheric pressure air

DEFF Research Database (Denmark)

Zhu, Jiajian; Gao, Jinlong; Li, Zhongshan

2014-01-01

Rapid transition from glow discharge to thermal arc has been a common problem in generating stable high-power non-thermal plasmas especially at ambient conditions. A sustained diffusive gliding arc discharge was generated in a large volume in atmospheric pressure air, driven by an alternating...... current (AC) power source. The plasma column extended beyond the water-cooled stainless steel electrodes and was stabilized by matching the flow speed of the turbulent air jet with the rated output power. Comprehensive investigations were performed using high-speed movies measured over the plasma column...

Brittle-ductile gliding shear zone and its dynamic metallization in uranium deposit No. 3110

International Nuclear Information System (INIS)

Fang Shiyi.

1990-01-01

A preliminary study on the macroscopic geological structure, microstructures of plastic deformation rotary strain, structural geochemistry and zoning regularity of a brittle-ductile gliding shear zone in uranium deposit No. 3110 is made. Structural dynamic metallization of uranium caused by the strong shearing stress is discussed. It is pointed out that great attention must be paid to in further exploration

In-situ determination of the mechanical properties of gliding or non-motile bacteria by atomic force microscopy under physiological conditions without immobilization.

Directory of Open Access Journals (Sweden)

Samia Dhahri

Full Text Available We present a study about AFM imaging of living, moving or self-immobilized bacteria in their genuine physiological liquid medium. No external immobilization protocol, neither chemical nor mechanical, was needed. For the first time, the native gliding movements of Gram-negative Nostoc cyanobacteria upon the surface, at speeds up to 900 µm/h, were studied by AFM. This was possible thanks to an improved combination of a gentle sample preparation process and an AFM procedure based on fast and complete force-distance curves made at every pixel, drastically reducing lateral forces. No limitation in spatial resolution or imaging rate was detected. Gram-positive and non-motile Rhodococcus wratislaviensis bacteria were studied as well. From the approach curves, Young modulus and turgor pressure were measured for both strains at different gliding speeds and are ranging from 20±3 to 105±5 MPa and 40±5 to 310±30 kPa depending on the bacterium and the gliding speed. For Nostoc, spatially limited zones with higher values of stiffness were observed. The related spatial period is much higher than the mean length of Nostoc nodules. This was explained by an inhomogeneous mechanical activation of nodules in the cyanobacterium. We also observed the presence of a soft extra cellular matrix (ECM around the Nostoc bacterium. Both strains left a track of polymeric slime with variable thicknesses. For Rhodococcus, it is equal to few hundreds of nanometers, likely to promote its adhesion to the sample. While gliding, the Nostoc secretes a slime layer the thickness of which is in the nanometer range and increases with the gliding speed. This result reinforces the hypothesis of a propulsion mechanism based, for Nostoc cyanobacteria, on ejection of slime. These results open a large window on new studies of both dynamical phenomena of practical and fundamental interests such as the formation of biofilms and dynamic properties of bacteria in real physiological conditions.

In-Situ Determination of the Mechanical Properties of Gliding or Non-Motile Bacteria by Atomic Force Microscopy under Physiological Conditions without Immobilization

Science.gov (United States)

Dhahri, Samia; Ramonda, Michel; Marlière, Christian

2013-01-01

We present a study about AFM imaging of living, moving or self-immobilized bacteria in their genuine physiological liquid medium. No external immobilization protocol, neither chemical nor mechanical, was needed. For the first time, the native gliding movements of Gram-negative Nostoc cyanobacteria upon the surface, at speeds up to 900 µm/h, were studied by AFM. This was possible thanks to an improved combination of a gentle sample preparation process and an AFM procedure based on fast and complete force-distance curves made at every pixel, drastically reducing lateral forces. No limitation in spatial resolution or imaging rate was detected. Gram-positive and non-motile Rhodococcus wratislaviensis bacteria were studied as well. From the approach curves, Young modulus and turgor pressure were measured for both strains at different gliding speeds and are ranging from 20±3 to 105±5 MPa and 40±5 to 310±30 kPa depending on the bacterium and the gliding speed. For Nostoc, spatially limited zones with higher values of stiffness were observed. The related spatial period is much higher than the mean length of Nostoc nodules. This was explained by an inhomogeneous mechanical activation of nodules in the cyanobacterium. We also observed the presence of a soft extra cellular matrix (ECM) around the Nostoc bacterium. Both strains left a track of polymeric slime with variable thicknesses. For Rhodococcus, it is equal to few hundreds of nanometers, likely to promote its adhesion to the sample. While gliding, the Nostoc secretes a slime layer the thickness of which is in the nanometer range and increases with the gliding speed. This result reinforces the hypothesis of a propulsion mechanism based, for Nostoc cyanobacteria, on ejection of slime. These results open a large window on new studies of both dynamical phenomena of practical and fundamental interests such as the formation of biofilms and dynamic properties of bacteria in real physiological conditions. PMID:23593493

Performance characteristics of the ARCHITECT anti-HCV assay.

Science.gov (United States)

Jonas, Gesa; Pelzer, Claudia; Beckert, Christian; Hausmann, Michael; Kapprell, Hans-Peter

2005-10-01

The ARCHITECT Anti-HCV assay is a fully automated high throughput chemiluminescent microparticle immunoassay (CMIA) for the detection of antibodies to structural and nonstructural proteins of the hepatitis C virus (HCV). To further enhance the performance of this test, the assay was modified to improve the specificity for blood donor specimens. The specificity of the enhanced ARCHITECT Anti-HCV assay was evaluated by screening blood donor samples randomly collected from various German blood banks, as well as hospitalized patient samples derived from Germany and the US. Additionally, antibody sensitivity was determined on commercially available anti-HCV seroconversion panels and on a commercially available worldwide anti-HCV genotype performance panel. Apparent specificity of the modified ARCHITECT Anti-HCV assay in a blood donor population consisting of 3811 specimens was 99.92%, compared to 99.76% for the current on-market assay. Additionally, antibody sensitivity was determined on commercially available anti-HCV seroconversion panels. Seroconversion sensitivity equivalent to or better than the current on-market product was observed by testing 33 seroconversion panels. This study demonstrates that the modified version of the ARCHITECT Anti-HCV assay shows improved specificity for blood donor specimens compared to the current assay on market without compromising sensitivity. With the availability of the improved ARCHITECT Anti-HCV assay and the recent launch of the ARCHITECT HIV Ag/Ab Combo assay, the ARCHITECT system now offers a full hepatitis/retrovirus menu with excellent performance on a high throughput, random access, automated analyzer, ideally suited for blood screening and diagnostic applications.

How-to-do-it: Immunological Assays for the Classroom 1. Enzyme Linked Immunosorbent Assay (ELISA): A Laboratory Tool for Demonstration of Antibody-Antigen Interaction.

Science.gov (United States)

Russo, A. J.; And Others

1984-01-01

Background information, list of required materials, and procedures are provided for an immunological assay which has been modified for use as a classroom/laboratory demonstration of antigen-antibody reaction. The assay is designed for a two and one-half hour laboratory period but may be modified for one hour laboratories. (JN)

Magnetic circuit design of magnetically driving gliding arc discharge device

International Nuclear Information System (INIS)

Jiang Zhonghe; Liu Minghai; Gu Chenglin; Pan Yuan

2002-01-01

A gliding arc discharge driven by magnetic field at atmospheric pressure can generate non-equilibrium plasma with good confinement property, and has extensive application in the areas of microelectronic fabrication, environmental engineering, etc. The magnetic circuit of the generator is designed with the permeance method, and analytic expression is obtained on the magnetic induction, the permeant magnetic material thickness and length of air gap. The results have been compared with those of the finite element method, the difference is 3.1%. But the permeance method is more concise and convenient and more universal and economical. So the permeance method is a more credible and useful engineering arithmetic

Instantaneous imaging of ozone in a gliding arc discharge using photofragmentation laser-induced fluorescence

Science.gov (United States)

Larsson, Kajsa; Hot, Dina; Gao, Jinlong; Kong, Chengdong; Li, Zhongshan; Aldén, Marcus; Bood, Joakim; Ehn, Andreas

2018-04-01

Ozone vapor, O3, is here visualized in a gliding arc discharge using photofragmentation laser-induced fluorescence. Ozone is imaged by first photodissociating the O3 molecule into an O radical and a vibrationally hot O2 fragment by a pump photon. Thereafter, the vibrationally excited O2 molecule absorbs a second (probe) photon that further transits the O2-molecule to an excited electronic state, and hence, fluorescence from the deexcitation process in the molecule can be detected. Both the photodissociation and excitation processes are achieved within one 248 nm KrF excimer laser pulse that is formed into a laser sheet and the fluorescence is imaged using an intensified CCD camera. The laser-induced signal in the vicinity of the plasma column formed by the gliding arc is confirmed to stem from O3 rather than plasma produced vibrationally hot O2. While both these products can be produced in plasmas a second laser pulse at 266 nm was utilized to separate the pump- from the probe-processes. Such arrangement allowed lifetime studies of vibrationally hot O2, which under these conditions were several orders of magnitude shorter than the lifetime of plasma-produced ozone.

Conversion of Methane to C2 Hydrocarbons and Hydrogen Using a Gliding Arc Reactor

International Nuclear Information System (INIS)

Hu Shuanghui; Wang Baowei; Lv Yijun; Yan Wenjuan

2013-01-01

Methane conversion has been studied using gliding arc plasma in the presence of argon. The process was conducted at atmospheric pressure and ambient temperature. The focus of this research was to develop a process of converting methane to C 2 hydrocarbons and hydrogen. The main parameters, including the CH 4 /Ar mole ratio, the CH 4 flow rate, the input voltage, and the minimum electrode gap, were varied to investigate their effects on methane conversion rate, product distribution, energy consumption, carbon deposit, and reaction stability. The specific energy requirement (SER) was used to express the energy utilization efficiency of the process and provided a practical guidance for optimizing reaction conditions for improving energy efficiency. It was found that the carbon deposition was not conducive to methane conversion, and the gliding arc plasma discharge reached a stable state twelve minutes later. Optimum conditions for methane conversion were suggested. The maximum methane conversion rate of 43.39% was obtained under the optimum conditions. Also, C 2 hydrocarbons selectivity, C 2 hydrocarbons yield, H 2 selectivity, H 2 yield and SER were 87.20%, 37.83%, 81.28%, 35.27%, and 2.09 MJ/mol, respectively.

Micro-CT evaluation of several glide path techniques and ProTaper Next shaping outcomes in maxillary first molar curved canals.

Science.gov (United States)

Alovisi, M; Cemenasco, A; Mancini, L; Paolino, D; Scotti, N; Bianchi, C C; Pasqualini, D

2017-04-01

To evaluate the ability of ProGlider instruments, PathFiles and K-files to maintain canal anatomy during glide path preparation using X-ray computed micro-tomography (micro-CT). Forty-five extracted maxillary first permanent molars were selected. Mesio-buccal canals were randomly assigned (n = 15) to manual K-file, PathFile or ProGlider groups for glide path preparation. Irrigation was achieved with 5% NaOCl and 10% EDTA. After glide path preparation, each canal was shaped with ProTaper Next X1 and X2 to working length. Specimens were scanned (isotropic voxel size 9.1 μm) for matching volumes and surface areas and post-treatment analyses. Canal volume, surface area, centroid shift, canal geometry variation through ratio of diameter ratios and ratio of cross-sectional areas were assessed in the apical and coronal levels and at the point of maximum canal curvature. One-way factorial anovas were used to evaluate the significance of instrument in the various canal regions. Post-glide path analysis revealed that instrument factor was significant at the apical level for both the ratio of diameter ratios and the ratio of cross-sectional areas (P flare the root canal compared with K-file and PathFile. PathFile and ProGlider demonstrated a significantly lower centroid shift compared with K-file at the apical level (P = 0.023). Post-shaping analysis demonstrated a more centred preparation of ProGlider, compared with PathFile and K-files, with no significant differences for other parameters. Use of ProGlider instruments led to less canal transportation than PathFiles and K-files. © 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Measurement of cetuximab and panitumumab-unbound serum EGFR extracellular domain using an assay based on slow off-rate modified aptamer (SOMAmer reagents.

Directory of Open Access Journals (Sweden)

Noh Jin Park

Full Text Available Response to cetuximab (Erbitux® and panitumumab (Vectibix® varies among individuals, and even those who show response ultimately gain drug resistance. One possible etiologic factor is differential interaction between the drug and target. We describe the development of an assay based on Slow Off-rate Modified Aptamer (SOMAmer(™ reagents that can distinguish drug-bound from unbound epidermal growth factor receptor (EGFR.This quantitative assay uses a SOMAmer reagent specific for EGFR extracellular domain (ECD as a capturing reagent. Captured SOMAmer is quantitated using PCR. Linearity and accuracy (recovery of the assay were assessed using normal sera and purified EGFR ECD.This EGFR ECD assay showed linearity between 2.5 and 600 ng/mL. Average recovery was 101%. The assay detected EGFR but showed little cross-reactivity to other ErbB proteins: 0.4% for ErbB2, 6.9% for ErbB3, and 1.3% for ErbB4. Preincubation of normal serum with either cetuximab or panitumumab resulted in a dose-dependent decrease in EGFR ECD levels measured using the SOMAmer assay; preincubation did not affect measurement with an ELISA.This SOMAmer-based serum EGFR ECD assay accurately and specifically measures EGFR in serum. Detection of significant amounts of drug-unbound EGFR in patients undergoing cetuximab or panitumumab treatment could be an indicator of poor drug response. Further studies are needed to evaluate the utility of the assay as an indicator of drug efficacy or as a guide to dosing.

Rate controlling mechanisms during hot deformation of Mg–3Gd–1Zn magnesium alloy: Dislocation glide and climb, dynamic recrystallization, and mechanical twinning

International Nuclear Information System (INIS)

Mirzadeh, H.; Roostaei, M.; Parsa, M.H.; Mahmudi, R.

2015-01-01

Highlights: • Hot deformation behavior and dynamic recrystallization of GZ31 magnesium alloy. • Deducing the operative deformation mechanisms by constitutive analysis. • Viscous glide as the rate controlling step during hot working of GZ31 alloy. • Characterization of the effect of mechanical twinning on constitutive relations. - Abstract: The flow behavior of the Mg–3Gd–1Zn (GZ31) magnesium alloy during hot working was critically analyzed and dislocation glide in the form of a viscous drag process (viscous glide) was identified as the rate controlling mechanism due to interaction of rare earth Gd atoms with the moving dislocations. Mechanical twinning was shown to significantly affect the level of flow stress at high Zener–Hollomon parameters, i.e. low forming temperatures and high strain rates. Moreover, dynamic recrystallization (DRX) was found to be another responsible phenomenon for deviation of constitutive equations from the theoretical ones, namely the deformation activation energy based on diffusivity and the pre-defined Garofalo’s type hyperbolic sine power, during high-temperature thermomechanical processing of this creep resistant light alloy

Differential Recognition of Pitch Patterns in Discrete and Gliding Stimuli in Congenital Amusia: Evidence from Mandarin Speakers

Science.gov (United States)

Liu, Fang; Xu, Yi; Patel, Aniruddh D.; Francart, Tom; Jiang, Cunmei

2012-01-01

This study examined whether "melodic contour deafness" (insensitivity to the direction of pitch movement) in congenital amusia is associated with specific types of pitch patterns (discrete versus gliding pitches) or stimulus types (speech syllables versus complex tones). Thresholds for identification of pitch direction were obtained using discrete…

Anatomy and histochemistry of spread-wing posture in birds. 2. Gliding flight in the California gull, Larus californicus: a paradox of fast fibers and posture.

Science.gov (United States)

Meyers, R A; Mathias, E

1997-09-01

Gliding flight is a postural activity which requires the wings to be held in a horizontal position to support the weight of the body. Postural behaviors typically utilize isometric contractions in which no change in length takes place. Due to longer actin-myosin interactions, slow contracting muscle fibers represent an economical means for this type of contraction. In specialized soaring birds, such as vultures and pelicans, a deep layer of the pectoralis muscle, composed entirely of slow fibers, is believed to perform this function. Muscles involved in gliding posture were examined in California gulls (Larus californicus) and tested for the presence of slow fibers using myosin ATPase histochemistry and antibodies. Surprisingly small numbers of slow fibers were found in the M. extensor metacarpi radialis, M. coracobrachialis cranialis, and M. coracobrachialis caudalis, which function in wrist extension, wing protraction, and body support, respectively. The low number of slow fibers in these muscles and the absence of slow fibers in muscles associated with wing extension and primary body support suggest that gulls do not require slow fibers for their postural behaviors. Gulls also lack the deep belly to the pectoralis found in other gliding birds. Since bird muscle is highly oxidative, we hypothesize that fast muscle fibers may function to maintain wing position during gliding flight in California gulls.

The Screw-Like Movement of a Gliding Bacterium Is Powered by Spiral Motion of Cell-Surface Adhesins.

Science.gov (United States)

Shrivastava, Abhishek; Roland, Thibault; Berg, Howard C

2016-09-06

Flavobacterium johnsoniae, a rod-shaped bacterium, glides over surfaces at speeds of ∼2 μm/s. The propulsion of a cell-surface adhesin, SprB, is known to enable gliding. We used cephalexin to generate elongated cells with irregular shapes and followed their displacement in three dimensions. These cells rolled about their long axes as they moved forward, following a right-handed trajectory. We coated gold nanoparticles with an SprB antibody and tracked them in three dimensions in an evanescent field where the nanoparticles appeared brighter when they were closer to the glass. The nanoparticles followed a right-handed spiral trajectory on the surface of the cell. Thus, if SprB were to adhere to the glass rather than to a nanoparticle, the cell would move forward along a right-handed trajectory, as observed, but in a direction opposite to that of the nanoparticle. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Evaluation of selected mechanical properties of NiTi rotary glide path files manufactured from controlled memory wires.

Science.gov (United States)

Nishijo, Miki; Ebihara, Arata; Tokita, Daisuke; Doi, Hisashi; Hanawa, Takao; Okiji, Takashi

2018-03-28

This study aimed to investigate mechanical properties related to flexibility and fracture resistance of controlled memory wiremanufactured nickel-titanium rotary glide path files [HyFlex EDM Glide Path File (EDM) and HyFlex GPF (GPF)]. Scout RaCe (RaCe) served as control. Bending loads, torsional/cyclic fatigue resistance, and screw-in forces were measured. EDM showed a significantly larger torque at fracture, a longer time to cyclic fracture in reciprocation and a larger screw-in force compared with GPF and RaCe. GPF showed significantly lower bending loads and higher angular deflection values than EDM and RaCe, and a significantly longer time to cyclic fracture than RaCe. The time to cyclic fracture was significantly longer in reciprocation compared with continuous rotation in EDM and GPF. It can be concluded that EDM and/or GPF showed higher flexibility and cyclic/torsional fatigue resistance compared with RaCe; and that reciprocation conferred better cyclic fatigue resistance to EDM and GPF.

Development of a colloidal gold immunochromatographic strip assay for simple and fast detection of human α-lactalbumin in genetically modified cow milk.

Science.gov (United States)

Tao, Chenyu; Zhang, Qingde; Feng, Na; Shi, Deshi; Liu, Bang

2016-03-01

The qualitative and quantitative declaration of food ingredients is important to consumers, especially for genetically modified food as it experiences a rapid increase in sales. In this study, we designed an accurate and rapid detection system using colloidal gold immunochromatographic strip assay (GICA) methods to detect genetically modified cow milk. First, we prepared 2 monoclonal antibodies for human α-lactalbumin (α-LA) and measured their antibody titers; the one with the higher titer was used for further experiments. Then, we found the optimal pH value and protein amount of GICA for detection of pure milk samples. The developed strips successfully detected genetically modified cow milk and non-modified cow milk. To determine the sensitivity of GICA, a quantitative ELISA system was used to determine the exact amount of α-LA, and then genetically modified milk was diluted at different rates to test the sensitivity of GICA; the sensitivity was 10 μg/mL. Our results demonstrated that the applied method was effective to detect human α-LA in cow milk. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Complete genome sequence of the gliding, heparinolytic Pedobacter saltans type strain (113T)

OpenAIRE

Liolios, Konstantinos; Sikorski, Johannes; Lu, Meagan; Nolan, Matt; Lapidus, Alla; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxanne; Han, Cliff; Goodwin, Lynne; Pitluck, Sam; Huntemann, Marcel; Ivanova, Natalia

2011-01-01

Pedobacter saltans Steyn et al. 1998 is one of currently 32 species in the genus Pedobacter within the family Sphingobacteriaceae. The species is of interest for its isolated location in the tree of life. Like other members of the genus P. saltans is heparinolytic. Cells of P. saltans show a peculiar gliding, dancing motility and can be distinguished from other Pedobacter strains by their ability to utilize glycerol and the inability to assimilate D-cellobiose. The genome presented here is on...

Functional Conservation of the Glide/Gcm Regulatory Network Controlling Glia, Hemocyte, and Tendon Cell Differentiation in Drosophila

Science.gov (United States)

Cattenoz, Pierre B.; Popkova, Anna; Southall, Tony D.; Aiello, Giuseppe; Brand, Andrea H.; Giangrande, Angela

2016-01-01

High-throughput screens allow us to understand how transcription factors trigger developmental processes, including cell specification. A major challenge is identification of their binding sites because feedback loops and homeostatic interactions may mask the direct impact of those factors in transcriptome analyses. Moreover, this approach dissects the downstream signaling cascades and facilitates identification of conserved transcriptional programs. Here we show the results and the validation of a DNA adenine methyltransferase identification (DamID) genome-wide screen that identifies the direct targets of Glide/Gcm, a potent transcription factor that controls glia, hemocyte, and tendon cell differentiation in Drosophila. The screen identifies many genes that had not been previously associated with Glide/Gcm and highlights three major signaling pathways interacting with Glide/Gcm: Notch, Hedgehog, and JAK/STAT, which all involve feedback loops. Furthermore, the screen identifies effector molecules that are necessary for cell-cell interactions during late developmental processes and/or in ontogeny. Typically, immunoglobulin (Ig) domain–containing proteins control cell adhesion and axonal navigation. This shows that early and transiently expressed fate determinants not only control other transcription factors that, in turn, implement a specific developmental program but also directly affect late developmental events and cell function. Finally, while the mammalian genome contains two orthologous Gcm genes, their function has been demonstrated in vertebrate-specific tissues, placenta, and parathyroid glands, begging questions on the evolutionary conservation of the Gcm cascade in higher organisms. Here we provide the first evidence for the conservation of Gcm direct targets in humans. In sum, this work uncovers novel aspects of cell specification and sets the basis for further understanding of the role of conserved Gcm gene regulatory cascades. PMID:26567182

Development and inter-laboratory assessment of droplet digital PCR assays for multiplex quantification of 15 genetically modified soybean lines.

Science.gov (United States)

Košir, Alexandra Bogožalec; Spilsberg, Bjørn; Holst-Jensen, Arne; Žel, Jana; Dobnik, David

2017-08-17

Quantification of genetically modified organisms (GMOs) in food and feed products is often required for their labelling or for tolerance thresholds. Standard-curve-based simplex quantitative polymerase chain reaction (qPCR) is the prevailing technology, which is often combined with screening analysis. With the rapidly growing number of GMOs on the world market, qPCR analysis becomes laborious and expensive. Innovative cost-effective approaches are therefore urgently needed. Here, we report the development and inter-laboratory assessment of multiplex assays to quantify GMO soybean using droplet digital PCR (ddPCR). The assays were developed to facilitate testing of foods and feed for compliance with current GMO regulations in the European Union (EU). Within the EU, the threshold for labelling is 0.9% for authorised GMOs per ingredient. Furthermore, the EU has set a technical zero tolerance limit of 0.1% for certain unauthorised GMOs. The novel multiplex ddPCR assays developed target 11 GMO soybean lines that are currently authorised, and four that are tolerated, pending authorisation in the EU. Potential significant improvements in cost efficiency are demonstrated. Performance was assessed for the critical parameters, including limits of detection and quantification, and trueness, repeatability, and robustness. Inter-laboratory performance was also determined on a number of proficiency programme and real-life samples.

A Middle Triassic thoracopterid from China highlights the evolutionary origin of overwater gliding in early ray-finned fishes.

Science.gov (United States)

Xu, Guang-Hui; Zhao, Li-Jun; Shen, Chen-Chen

2015-01-01

Gliding adaptations in thoracopterid flying fishes represent a remarkable case of convergent evolution of overwater gliding strategy with modern exocoetid flying fishes, but the evolutionary origin of this strategy was poorly known in the thoracopterids because of lack of transitional forms. Until recently, all thoracopterids, from the Late Triassic of Austria and Italy and the Middle Triassic of South China, were highly specialized 'four-winged' gliders in having wing-like paired fins and an asymmetrical caudal fin with the lower caudal lobe notably larger than the upper lobe. Here, we show that the new genus Wushaichthys and the previously alleged 'peltopleurid' Peripeltopleurus, from the Middle Triassic (Ladinian, 235-242 Ma) of South China and near the Ladinian/Anisian boundary of southern Switzerland and northern Italy, respectively, represent the most primitive and oldest known thoracopterids. Wushaichthys, the most basal thoracopterid, shows certain derived features of this group in the skull. Peripeltopleurus shows a condition intermediate between Wushaichthys and Thoracopterus in having a slightly asymmetrical caudal fin but still lacking wing-like paired fins. Phylogenetic studies suggest that the evolution of overwater gliding of thoracopterids was gradual in nature; a four-stage adaption following the 'cranial specialization-asymmetrical caudal fin-enlarged paired fins-scale reduction' sequence has been recognized in thoracopterid evolution. Moreover, Wushaichthys and Peripeltopleurus bear hooklets on the anal fin of supposed males, resembling those of modern viviparious teleosts. Early thoracopterids probably had evolved a live-bearing reproductive strategy. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Cyclic fatigue resistance of R-Pilot, HyFlex EDM and PathFile nickel-titanium glide path files in artificial canals with double (S-shaped) curvature.

Science.gov (United States)

Uslu, G; Özyürek, T; Yılmaz, K; Gündoğar, M

2018-05-01

To examine the cyclic fatigue resistances of R-Pilot, HyFlex EDM and PathFile NiTi glide path files in S-shaped artificial canals. Twenty R-Pilot (12.5/.04), 20 HyFlex EDM (10/.05) and 20 PathFile (19/.02) single-file glide path files were included. Sixty files (n: 20/each) were subjected to static cyclic fatigue testing using double-curved canals until fracture occurred (TF). The number of cycles to fracture (NCF) was calculated by multiplying the rpm value by the TF. The length of the fractured fragment (FL) was determined by a digital microcaliper. Six samples of fractured files (n: 2/each) were examined by SEM to determine the fracture mode. The NCF and the FL data were analysed using one-way anova, post hoc Tamhane and Kruskal-Wallis tests using SPPS 21 software. The significance level was set at 5%. In the double-curved canal, all the files fractured first in the apical curvature and then in the coronal curvature. The NCF values revealed that the R-Pilot had the greatest cyclic fatigue resistance, followed by the HyFlex EDM and PathFile in both the apical and coronal curvatures (P < 0.05). R-Pilot NiTi glide path files, used in a reciprocating motion, had the greatest cyclic fatigue resistance amongst the tested NiTi glide path files in an artificial S-shaped canal. © 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Asymmetric, compressive, SiGe epilayers on Si grown by lateral liquid-phase epitaxy utilizing a distinction between dislocation nucleation and glide critical thicknesses

Science.gov (United States)

O'Reilly, Andrew J.; Quitoriano, Nathaniel

2018-01-01

Uniaxially strained Si1-xGex channels have been proposed as a solution for high mobility channels in next-generation MOSFETS to ensure continued device improvement as the benefits from further miniaturisation are diminishing. Previously proposed techniques to deposit uniaxially strained Si1-xGex epilayers on Si (0 0 1) substrates require multiple deposition steps and only yielded thin strips of uniaxially strained films. A lateral liquid-phase epitaxy (LLPE) technique was developed to deposit a blanket epilayer of asymmetrically strained Si97.4Ge2.6 on Si in a single step, where the epilayer was fully strained in the growth direction and 31% strain-relaxed in the orthogonal direction. The LLPE technique promoted the glide of misfit dislocations, which nucleated in a region with an orthogonal misfit dislocation network, into a region where the dislocation nucleation was inhibited. This created an array of parallel misfit dislocations which were the source of the asymmetric strain. By observing the thicknesses at which the dislocation network transitions from orthogonal to parallel and at which point dislocation glide is exhausted, the separate critical thicknesses for dislocation nucleation and dislocation glide can be determined.

CO2 Dissociation by Low Current Gliding Discharge in the Reverse Vortex Flow

Science.gov (United States)

Gutsol, Alexander

2012-10-01

If performed with high energy efficiency, plasma-chemical dissociation of carbon dioxide can be a way of converting and storing energy when there is an excess of electric energy, for example generated by solar elements of wind turbines. CO2 dissociation with efficiency of up to 90% was reported earlier for low pressure microwave discharge in supersonic flow. A new plasma-chemical system uses a low current gliding discharge in the reverse vortex flow of plasma gas. The system is a development of the Gliding Arc in Tornado reactor. The system was used to study dissociation of CO2 in wide ranges of the following experimental parameters: reactor pressure (15-150 kPa), discharge current (50-500 mA), gas flow rate (3-30 liters per minute), and electrode gap length (1-10 cm). Additionally, the effect of thermal energy recuperation on CO2 dissociation efficiency was tested. Plasma chemical efficiency of CO2 dissociation is very low (about 3%) in a short discharge at low pressures (about 15 kPa) when it is defined by electronic excitation. The highest efficiency (above 40%) was reached at pressures 50-70 kPa in a long discharge with thermal energy recuperation. It means that the process is controlled by thermal dissociation with subsequent effective quenching. Plasma chemical efficiency was determined from the data of chromatographic analysis and oscilloscope electric power integration, and also was checked calorimetrically by the thermal balance of the system.

A new stem-neopterygian fish from the Middle Triassic of China shows the earliest over-water gliding strategy of the vertebrates.

Science.gov (United States)

Xu, Guang-Hui; Zhao, Li-Jun; Gao, Ke-Qin; Wu, Fei-Xiang

2013-01-07

Flying fishes are extraordinary aquatic vertebrates capable of gliding great distances over water by exploiting their enlarged pectoral fins and asymmetrical caudal fin. Some 50 species of extant flying fishes are classified in the Exocoetidae (Neopterygii: Teleostei), which have a fossil record no older than the Eocene. The Thoracopteridae is the only pre-Cenozoic group of non-teleosts that shows an array of features associated with the capability of over-water gliding. Until recently, however, the fossil record of the Thoracopteridae has been limited to the Upper Triassic of Austria and Italy. Here, we report the discovery of exceptionally well-preserved fossils of a new thoracopterid flying fish from the Middle Triassic of China, which represents the earliest evidence of an over-water gliding strategy in vertebrates. The results of a phylogenetic analysis resolve the Thoracopteridae as a stem-group of the Neopterygii that is more crown-ward than the Peltopleuriformes, yet more basal than the Luganoiiformes. As the first record of the Thoracopteride in Asia, this new discovery extends the geographical distribution of this group from the western to eastern rim of the Palaeotethys Ocean, providing new evidence to support the Triassic biological exchanges between Europe and southern China. Additionally, the Middle Triassic date of the new thoracopterid supports the hypothesis that the re-establishment of marine ecosystems after end-Permian mass extinction is more rapid than previously thought.

Glitter-like iridescence within the bacteroidetes especially Cellulophaga spp.: optical properties and correlation with gliding motility.

Directory of Open Access Journals (Sweden)

Betty Kientz

Full Text Available Iridescence results from structures that generate color. Iridescence of bacterial colonies has recently been described and illustrated. The glitter-like iridescence class, created especially for a few strains of Cellulophaga lytica, exhibits an intense iridescence under direct illumination. Such color appearance effects were previously associated with other bacteria from the Bacteroidetes phylum, but without clear elucidation and illustration. To this end, we compared various bacterial strains to which the iridescent trait was attributed. All Cellulophaga species and additional Bacteroidetes strains from marine and terrestrial environments were investigated. A selection of bacteria, mostly marine in origin, were found to be iridescent. Although a common pattern of reflected wavelengths was recorded for the species investigated, optical spectroscopy and physical measurements revealed a range of different glitter-like iridescence intensity and color profiles. Importantly, gliding motility was found to be a common feature of all iridescent colonies. Dynamic analyses of "glitter" formation at the edges of C. lytica colonies showed that iridescence was correlated with layer superposition. Both gliding motility, and unknown cell-to-cell communication processes, may be required for the establishment, in time and space, of the necessary periodic structures responsible for the iridescent appearance of Bacteroidetes.

Employment of hypersonic glide vehicles: Proposed criteria for use

Energy Technology Data Exchange (ETDEWEB)

Olguin, Abel [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States)

2014-07-01

Hypersonic Glide Vehicles (HGVs) are a type of reentry vehicle that couples the high speed of ballistic missiles with the maneuverability of aircraft. The HGV has been in development since the 1970s, and its technology falls under the category of Conventional Prompt Global Strike (CPGS) weapons. As noted by James M. Acton, a senior associate in the Nuclear Policy Program at the Carnegie Endowment, CPGS is a “missile in search of a mission.” With the introduction of any significant new military capability, a doctrine for use—including specifics regarding how, when and where it would be used, as well as tactics, training and procedures—must be clearly defined and understood by policy makers, military commanders, and planners. In this paper, benefits and limitations of the HGV are presented. Proposed criteria and four scenarios illustrate a possible method for assessing when to use an HGV.

Aptamer-Phage Reporters for Ultrasensitive Lateral Flow Assays.

Science.gov (United States)

Adhikari, Meena; Strych, Ulrich; Kim, Jinsu; Goux, Heather; Dhamane, Sagar; Poongavanam, Mohan-Vivekanandan; Hagström, Anna E V; Kourentzi, Katerina; Conrad, Jacinta C; Willson, Richard C

2015-12-01

We introduce the modification of bacteriophage particles with aptamers for use as bioanalytical reporters, and demonstrate the use of these particles in ultrasensitive lateral flow assays. M13 phage displaying an in vivo biotinylatable peptide (AviTag) genetically fused to the phage tail protein pIII were used as reporter particle scaffolds, with biotinylated aptamers attached via avidin-biotin linkages, and horseradish peroxidase (HRP) reporter enzymes covalently attached to the pVIII coat protein. These modified viral nanoparticles were used in immunochromatographic sandwich assays for the direct detection of IgE and of the penicillin-binding protein from Staphylococcus aureus (PBP2a). We also developed an additional lateral flow assay for IgE, in which the analyte is sandwiched between immobilized anti-IgE antibodies and aptamer-bearing reporter phage modified with HRP. The limit of detection of this LFA was 0.13 ng/mL IgE, ∼100 times lower than those of previously reported IgE assays.

[fs-Lentotomy: presbyopia reversal by generating gliding planes inside the crystalline lens].

Science.gov (United States)

Lubatschowski, H; Schumacher, S; Wegener, A; Fromm, M; Oberheide, U; Hoffmann, H; Gerten, G

2009-12-01

Based on the Helmholtz theory for accommodation, increasing sclerosis of the lens nucleus and cortex is the main cause for the development of presbyopia. Existing therapies, however, do not reverse the stiffness of the crystalline lens and thus do not regain real accommodation ability. A new approach to restore the flexibility of the lens has been realised by utilising the non-linear interaction of ultrafast laser pulses with transparent tissue, the so-called photodisruption. This process has been used to create micro-incisions which act as gliding planes inside the crystalline lens without opening the eye globe. This treatment method, known as fs-lentotomy, enables regeneration of real dynamic accommodation. For the first time, 3D structures for gliding planes were successfully generated in experiments with human donor lenses of different ages. An average increase in anterior-posterior lens thickness of 100 mum accompanied by a decrease of equatorial lens diameter was observed as a direct consequence of fs-lentotomy. This is attributed to the increased flexibility, as the force of the capsule bag moulds the lens tissue more spherically. Moreover, in vivo experiments on rabbit eye lenses did not induce an increasing opacification (cataract) over a six-month follow-up period. However, the incisions were still detectable using Scheimpflug imaging and histopathological techniques, although the visibility of the incisions was declining. Furthermore, no side effects were observed during the wound healing process and during a six-months follow-up period. Based on these findings fs-lentotomy might have the potential to become a procedure for the reversal of presbyopia. Copyright Georg Thieme Verlag KG Stuttgart . New York.

Dislocation glide in Ni-Al solid solutions from the atomic scale up: a molecular dynamics study; Etude du glissement des dislocations dans la solution solide Ni-Al par simulation a l'echelle atomique

Energy Technology Data Exchange (ETDEWEB)

Rodary, E

2003-01-01

The glide of an edge dislocation in solid solutions is studied by molecular dynamics, at fixed temperature and imposed external stress. We have optimized an EAM potential for Ni(1 a 8% A1): it well reproduces the lattice expansion, local atomic order, stacking fault energy as a function of composition, as well as the elastic properties of the {gamma}' phase with L1{sub 2} structure. On increasing the stress, the dislocation is first immobile, then glides with a velocity proportional to the stress and the velocity saturates on reaching the transverse sound velocity. However, only beyond a static threshold stress, {sigma}{sub s}, does the dislocation glide a distance large enough to allow macroscopic shear; the linear part of the velocity-stress curve extrapolates to zero at a dynamical threshold stress, {sigma}{sub d}, The friction coefficient, and the threshold stresses ({sigma}{sub s} and {sigma}{sub d}), increase with the A1 concentration and decrease with temperature (300 and 500 K). Close to the critical shear stress, {sigma}{sub s}, the dislocation glide is analysed with a 'stop and go' model. The latter yields the flight velocity between obstacles, the mean obstacle density and the distribution of the waiting time on each obstacle as a function of stress, composition and temperature. The obstacle to the glide is proposed to be the strong repulsion between Al atoms brought into nearest neighbour position by the glide process, and not the dislocation-solute interaction. The microscopic parameters so defined are introduced into a micro-mechanical model, which well reproduces the known behaviour of nickel base solid solutions. (author)

Assay method and compositions

International Nuclear Information System (INIS)

1977-01-01

Methods are described for measuring catecholamine levels in human and animal body fluids and tissues using the catechol-O-methyl-transferase (COMT) radioassay. The assay involves incubating the biological sample with COMT and the tritiated methyl donor, S-adenosyl-L-methionine( 3 H)-methyl. The O-methylated ( 3 H) epinephrine and/or norepinephrine are extracted and oxidised to vanillin- 3 H which in turn is extracted and its radioactivity counted. When analysing dopamine levels the assay is extended by vanillin- 3 H and raising the pH of the aqueous periodate phase from which O-methylated ( 3 H) dopamine is extracted and counted. The assay may be modified depending on whether measurements of undifferentiated total endogenous catecholamine levels or differential analyses of the catecholamine levels are being performed. The sensitivity of the assay can be as low as 5 picograms for norepinephrine and epinephrine and 12 picograms for dopamine. The assemblance of the essential components of the assay into a kit for use in laboratories is also described. (U.K.)

Sensitive colorimetric assay for uric acid and glucose detection based on multilayer-modified paper with smartphone as signal readout.

Science.gov (United States)

Wang, Xu; Li, Fang; Cai, Ziqi; Liu, Kaifan; Li, Jing; Zhang, Boyang; He, Jianbo

2018-04-01

In this work, a multilayer-modified paper-based colorimetric sensing platform with improved color uniformity and intensity was developed for the sensitive and selective determination of uric acid and glucose with smartphone as signal readout. In detail, chitosan, different kinds of chromogenic reagents, and horseradish peroxidase (HRP) combined with a specific oxidase, e.g., uricase or glucose oxidase (GOD), were immoblized onto the paper substrate to form a multilayer-modified test paper. Hydrogen peroxide produced by the oxidases (uricase or GOD) reacts with the substrates (uric acid or glucose), and could oxidize the co-immoblized chromogenic reagents to form colored products with HRP as catalyst. A simple strategy by placing the test paper on top of a light-emitting diode lamp was adopted to efficiently prevent influence from the external light. The color images were recorded by the smartphone camera, and then the gray values of the color images were calculated for quantitative analysis. The developed method provided a wide linear response from 0.01 to 1.0 mM for uric acid detection and from 0.02 to 4.0 mM for glucose detection, with a limit of detection (LOD) as low as 0.003 and 0.014 mM, respectively, which was much lower than for previously reported paper-based colorimetric assays. The proposed assays were successfully applied to uric acid and glucose detection in real serum samples. Furthermore, the enhanced analytical performance of the proposed method allowed the non-invasive detection of glucose levels in tear samples, which holds great potential for point-of-care analysis. Graphical abstract ᅟ.

Development of a novel genetically modified bioluminescent-bacteria-based assay for detection of fluoroquinolones in animal-derived foods.

Science.gov (United States)

Cheng, Guyue; Dong, Xiaobing; Wang, Yulian; Peng, Dapeng; Wang, Xu; Hao, Haihong; Xie, Shuyu; Qu, Wei; Liu, Zhenli; Yuan, Zonghui

2014-12-01

Fluoroquinolones (FQNs) are broad-spectrum antibacterial agents widely used in animal husbandry and aquaculture. The residues and antimicrobial resistance of such antibiotics are a major public health concern. To realize multianalyte detection of FQN residues, a genetically modified bacterium, Escherichia coli pK12 harboring plasmid pRecAlux3, was constructed in this study to develop a bioluminescent-bacteria-based assay for the detection of FQNs in animal-derived foods. This assay was based on the principle of induction of an SOS response by FQNs via inducing the recA-promoter-fused luciferase reporter gene existing on the plasmid pRecAlux3. E. coli pK12 was able to recognize 11 FQNs: difloxacin, enrofloxacin, ciprofloxacin, sarafloxacin, norfloxacin, danofloxacin, ofloxacin, pefloxacin, lomefloxacin, marbofloxacin, and orbifloxacin. This method could be applied to 11 edible tissues, including milk, fish muscle, and the muscles, livers, and kidneys of cattle, chickens, and pigs, with a very simple and rapid sample extraction procedure using only phosphate-buffered saline. The limits of detection of the FQNs were between 12.5 and 100 μg kg(-1), all of which were lower than the maximum residue limits. Most of the recoveries of the FQNs were in the range from 60 to 120 %, and the interassay coefficients of variation were less than 30 %. This method, confirmed by high-performance liquid chromatography, is reliable and can be used as both a screening test and a semiquantitative assay, when the identity of a single type of FQN is known.

A versatile class of cell surface directional motors gives rise to gliding motility and sporulation in Myxococcus xanthus.

Directory of Open Access Journals (Sweden)

Morgane Wartel

2013-12-01

Full Text Available Eukaryotic cells utilize an arsenal of processive transport systems to deliver macromolecules to specific subcellular sites. In prokaryotes, such transport mechanisms have only been shown to mediate gliding motility, a form of microbial surface translocation. Here, we show that the motility function of the Myxococcus xanthus Agl-Glt machinery results from the recent specialization of a versatile class of bacterial transporters. Specifically, we demonstrate that the Agl motility motor is modular and dissociates from the rest of the gliding machinery (the Glt complex to bind the newly expressed Nfs complex, a close Glt paralogue, during sporulation. Following this association, the Agl system transports Nfs proteins directionally around the spore surface. Since the main spore coat polymer is secreted at discrete sites around the spore surface, its transport by Agl-Nfs ensures its distribution around the spore. Thus, the Agl-Glt/Nfs machineries may constitute a novel class of directional bacterial surface transporters that can be diversified to specific tasks depending on the cognate cargo and machinery-specific accessories.

Distance-Based Tear Lactoferrin Assay on Microfluidic Paper Device Using Interfacial Interactions on Surface-Modified Cellulose.

Science.gov (United States)

Yamada, Kentaro; Henares, Terence G; Suzuki, Koji; Citterio, Daniel

2015-11-11

"Distance-based" detection motifs on microfluidic paper-based analytical devices (μPADs) allow quantitative analysis without using signal readout instruments in a similar manner to classical analogue thermometers. To realize a cost-effective and calibration-free distance-based assay of lactoferrin in human tear fluid on a μPAD not relying on antibodies or enzymes, we investigated the fluidic mobilities of the target protein and Tb(3+) cations used as the fluorescent detection reagent on surface-modified cellulosic filter papers. Chromatographic elution experiments in a tear-like sample matrix containing electrolytes and proteins revealed a collapse of attractive electrostatic interactions between lactoferrin or Tb(3+) and the cellulosic substrate, which was overcome by the modification of the paper surface with the sulfated polysaccharide ι-carrageenan. The resulting μPAD based on the fluorescence emission distance successfully analyzed 0-4 mg mL(-1) of lactoferrin in complex human tear matrix with a lower limit of detection of 0.1 mg mL(-1) by simple visual inspection. Assay results of 18 human tear samples including ocular disease patients and healthy volunteers showed good correlation to the reference ELISA method with a slope of 0.997 and a regression coefficient of 0.948. The distance-based quantitative signal and the good batch-to-batch fabrication reproducibility relying on printing methods enable quantitative analysis by simply reading out "concentration scale marks" printed on the μPAD without performing any calibration and using any signal readout instrument.

More Than Gliding: Involvement of GldD and GldG in the Virulence of Flavobacterium psychrophilum

Directory of Open Access Journals (Sweden)

David Pérez-Pascual

2017-11-01

Full Text Available A fascinating characteristic of most members of the genus Flavobacterium is their ability to move over surfaces by gliding motility. Flavobacterium psychrophilum, an important pathogen of farmed salmonids worldwide, contains in its genome the 19 gld and spr genes shown to be required for gliding or spreading in Flavobacterium johnsoniae; however, their relative role in its lifestyle remains unknown. In order to address this issue, two spreading deficient mutants were produced as part of a Tn4351 mutant library in F. psychrophilum strain THCO2-90. The transposons were inserted in gldD and gldG genes. While the wild-type strain is proficient in adhesion, biofilm formation and displays strong proteolytic activity, both mutants lost these characteristics. Extracellular proteome comparisons revealed important modifications for both mutants, with a significant reduction of the amounts of proteins likely transported through the outer membrane by the Type IX secretion system, indicating that GldD and GldG proteins are required for an effective activity of this system. In addition, a significant decrease in virulence was observed using rainbow trout bath and injection infection models. Our results reveal additional roles of gldD and gldG genes that are likely of importance for the F. psychrophilum lifestyle, including virulence.

Glide back booster wind tunnel model testing

Science.gov (United States)

Pricop, M. V.; Cojocaru, M. G.; Stoica, C. I.; Niculescu, M. L.; Neculaescu, A. M.; Persinaru, A. G.; Boscoianu, M.

2017-07-01

Affordable space access requires partial or ideally full launch vehicle reuse, which is in line with clean environment requirement. Although the idea is old, the practical use is difficult, requiring very large technology investment for qualification. Rocket gliders like Space Shuttle have been successfullyoperated but the price and correspondingly the energy footprint were found not sustainable. For medium launchers, finally there is a very promising platform as Falcon 9. For very small launchers the situation is more complex, because the performance index (payload to start mass) is already small, versus medium and heavy launchers. For partial reusable micro launchers this index is even smaller. However the challenge has to be taken because it is likely that in a multiyear effort, technology is going to enable the performance recovery to make such a system economically and environmentally feasible. The current paper is devoted to a small unitary glide back booster which is foreseen to be assembled in a number of possible configurations. Although the level of analysis is not deep, the solution is analyzed from the aerodynamic point of view. A wind tunnel model is designed, with an active canard, to enablea more efficient wind tunnel campaign, as a national level premiere.

Developing a yeast-based assay protocol to monitor total ...

African Journals Online (AJOL)

A yeast-based assay protocol developed for detecting oestrogenic activity in activated sludge (AS) supernatant is described. The protocol used Saccharomyces cerevisiae construct RMY/ER-ERE with human oestrogen receptor (ERα) and lacZ reporter genes, and was developed by modifying existing assays for use with AS ...

Unsaturated fatty acids show clear elicitation responses in a modified local lymph node assay with an elicitation phase, and test positive in the direct peptide reactivity assay.

Science.gov (United States)

Yamashita, Kunihiko; Shinoda, Shinsuke; Hagiwara, Saori; Miyazaki, Hiroshi; Itagaki, Hiroshi

2015-12-01

The Organisation for Economic Co-operation and Development (OECD) Test Guidelines (TG) adopted the murine local lymph node assay (LLNA) and guinea pig maximization test (GPMT) as stand-alone skin sensitization test methods. However, unsaturated carbon-carbon double-bond and/or lipid acids afforded false-positive results more frequently in the LLNA compared to those in the GPMT and/or in human subjects. In the current study, oleic, linoleic, linolenic, undecylenic, fumaric, maleic, and succinic acid and squalene were tested in a modified LLNA with an elicitation phase (LLNA:DAE), and in a direct peptide reactivity assay (DPRA) to evaluate their skin-sensitizing potential. Oleic, linoleic, linolenic, undecylenic and maleic acid were positive in the LLNA:DAE, of which three, linoleic, linolenic, and maleic acid were positive in the DPRA. Furthermore, the results of the cross-sensitizing tests using four LLNA:DAE-positive chemicals were negative, indicating a chemical-specific elicitation response. In a previous report, the estimated concentration needed to produce a stimulation index of 3 (EC3) of linolenic acid, squalene, and maleic acid in the LLNA was LLNA. However, the skin-sensitizing potential of all LLNA:DAE-positive chemicals was estimated as weak. These results suggested that oleic, linoleic, linolenic, undecylenic, and maleic acid had skin-sensitizing potential, and that the LLNA overestimated the skin-sensitizing potential compared to that estimated by the LLNA:DAE.

Characterization and evaluation of a modified local lymph node assay using ATP content as a non-radio isotopic endpoint.

Science.gov (United States)

Idehara, Kenji; Yamagishi, Gaku; Yamashita, Kunihiko; Ito, Michio

2008-01-01

The murine local lymph node assay (LLNA) is an accepted and widely used method for assessing the skin-sensitizing potential of chemicals. Here, we describe a non-radio isotopic modified LLNA in which adenosine triphosphate (ATP) content is used as an endpoint instead of radioisotope (RI); the method is termed LLNA modified by Daicel based on ATP content (LLNA-DA). Groups of female CBA/JNCrlj mice were treated topically on the dorsum of both ears with test chemicals or a vehicle control on days 1, 2, and 3; an additional fourth application was conducted on day 7. Pretreatment with 1% sodium lauryl sulfate solution was performed 1 h before each application. On day 8, the amount of ATP in the draining auricular lymph nodes was measured as an alternative endpoint by the luciferin-luciferase assay in terms of bioluminescence (relative light units, RLU). A stimulation index (SI) relative to the concurrent vehicle control was derived based on the RLU value, and an SI of 3 was set as the cut-off value. Using the LLNA-DA method, 31 chemicals were tested and the results were compared with those of other test methods. The accuracy of LLNA-DA vs LLNA, guinea pig tests, and human tests was 93% (28/30), 80% (20/25), and 79% (15/19), respectively. The estimated concentration (EC) 3 value was calculated and compared with that of the original LLNA. It was found that the EC3 values obtained by LLNA-DA were almost equal to those obtained by the original LLNA. The SI value based on ATP content is similar to that of the original LLNA as a result of the modifications in the chemical treatment procedure, which contribute to improving the SI value. It is concluded that LLNA-DA is a promising non-RI alternative method for evaluating the skin-sensitizing potential of chemicals.

In Vivo Alkaline Comet Assay and Enzyme-modified Alkaline Comet Assay for Measuring DNA Strand Breaks and Oxidative DNA Damage in Rat Liver.

Science.gov (United States)

Ding, Wei; Bishop, Michelle E; Lyn-Cook, Lascelles E; Davis, Kelly J; Manjanatha, Mugimane G

2016-05-04

Unrepaired DNA damage can lead to genetic instability, which in turn may enhance cancer development. Therefore, identifying potential DNA damaging agents is important for protecting public health. The in vivo alkaline comet assay, which detects DNA damage as strand breaks, is especially relevant for assessing the genotoxic hazards of xenobiotics, as its responses reflect the in vivo absorption, tissue distribution, metabolism and excretion (ADME) of chemicals, as well as DNA repair process. Compared to other in vivo DNA damage assays, the assay is rapid, sensitive, visual and inexpensive, and, by converting oxidative DNA damage into strand breaks using specific repair enzymes, the assay can measure oxidative DNA damage in an efficient and relatively artifact-free manner. Measurement of DNA damage with the comet assay can be performed using both acute and subchronic toxicology study designs, and by integrating the comet assay with other toxicological assessments, the assay addresses animal welfare requirements by making maximum use of animal resources. Another major advantage of the assays is that they only require a small amount of cells, and the cells do not have to be derived from proliferating cell populations. The assays also can be performed with a variety of human samples obtained from clinically or occupationally exposed individuals.

Non-planar dislocations: 3D models and thermally-activated glide processes

International Nuclear Information System (INIS)

Ngan, A.H.W.

2005-01-01

In recent years, there has been a renewed interest in studying the cross-slip of screw dislocations in the simple face-centred cubic (FCC) structure. This paper serves to address parallel developments in modelling the cross-slip of screw dislocations in the body-centred cubic (BCC) structure and the ordered L1 2 structure. In the latter two cases, the dislocation cores have non-planar spreading offering high intrinsic Peierls stresses. The flow behaviours of these materials, such as the non-Schmid behaviour and temperature-dependence of flow stress, are largely due to the behaviours of single dislocations. 3D atomistic modelling of the minimum-energy path for the glide processes in these cases is performed with an aim to reconcile with experimentally determined activation energies for slip

In vitro digestibility of individual amino acids in rumen-undegraded protein: the modified three-step procedure and the immobilized digestive enzyme assay.

Science.gov (United States)

Boucher, S E; Calsamiglia, S; Parsons, C M; Stern, M D; Moreno, M Ruiz; Vázquez-Añón, M; Schwab, C G

2009-08-01

Three soybean meal, 3 SoyPlus (West Central Cooperative, Ralston, IA), 5 distillers dried grains with solubles, and 5 fish meal samples were used to evaluate the modified 3-step in vitro procedure (TSP) and the in vitro immobilized digestive enzyme assay (IDEA; Novus International Inc., St. Louis, MO) for estimating digestibility of AA in rumen-undegraded protein (RUP-AA). In a previous experiment, each sample was ruminally incubated in situ for 16 h, and in vivo digestibility of AA in the intact samples and in the rumen-undegraded residues (RUR) was obtained for all samples using the precision-fed cecectomized rooster assay. For the modified TSP, 5 g of RUR was weighed into polyester bags, which were then heat-sealed and placed into Daisy(II) incubator bottles. Samples were incubated in a pepsin/HCl solution followed by incubation in a pancreatin solution. After this incubation, residues remaining in the bags were analyzed for AA, and digestibility of RUP-AA was calculated based on disappearance from the bags. In vitro RUP-AA digestibility estimates obtained with this procedure were highly correlated to in vivo estimates. Corresponding intact feeds were also analyzed via the pepsin/pancreatin steps of the modified TSP. In vitro estimates of AA digestibility of the feeds were highly correlated to in vivo RUP-AA digestibility, which suggests that the feeds may not need to be ruminally incubated before determining RUP-AA digestibility in vitro. The RUR were also analyzed via the IDEA kits. The IDEA values of the RUR were good predictors of RUP-AA digestibility in soybean meal, SoyPlus, and distillers dried grains with solubles, but the IDEA values were not as good predictors of RUP-AA digestibility in fish meal. However, the IDEA values of intact feed samples were also determined and were highly correlated to in vivo RUP-AA digestibility for all feed types, suggesting that the IDEA value of intact feeds may be a better predictor of RUP-AA digestibility than the IDEA

Sensitive, specific radioisotope assay for L-glutamine-D-fructose-6- phosphat aminotransferase

International Nuclear Information System (INIS)

Callahan, M.; Tourian, A.; Hung, W.Y.

1981-01-01

A sensitive and specific radioassay for L-glutamine-D-fructose-6-phosphate aminotransferase (EC 5.3.1.19) activity is presented. Picomoles of product are measurable, and the assay can be applied to systems having limited quantities of available protein, particularly in extracts of either cell or organ cultures. The assay is at least 10,000 times more sensitive under K 1 concentrations of fructose 6-phosphate than the modified Elson-Morgan colorimetric assay and 20 times more sensitive under saturating conditions of fructose 6-phosphate. As little as 0.5 μg of cell-extract protein will yield measurable product. In contrast, 280 μg of crude-extract protein from colon is required with the modified Elson-Morgan colorimetric assay

Validation of a modified fluorimetric assay for the screening of trichomonacidal drugs

Directory of Open Access Journals (Sweden)

Alexandra Ibáñez Escribano

2012-08-01

Full Text Available A fluorimetric microassay that uses a redox dye to determine the viability of the flagellate Trichomonas vaginalis has been optimised to provide a more sensitive method to evaluate potential trichomonacidal compounds. Resazurin has been used in recent years to test drugs against different parasites, including trichomonadid protozoa; however, the reproducibility of these resazurin-based methods in our laboratory has been limited because the flagellate culture medium spontaneously reduces the resazurin. The objective of this work was to refine the fluorimetric microassay method previously developed by other research groups to reduce the fluorescence background generated by the media and increase the sensitivity of the screening assay. The experimental conditions, time of incubation, resazurin concentration and media used in the microtitre plates were adjusted. Different drug sensitivity studies against T. vaginalis were developed using the 5-nitroimidazole reference drugs, new 5-nitroindazolinones and 5-nitroindazole synthetic derivatives. Haemocytometer count results were compared with the resazurin assay using a 10% solution of 3 mM resazurin dissolved in phosphate buffered saline with glucose (1 mg/mL. The fluorimetric assay and the haemocytometer counts resulted in similar percentages of trichomonacidal activity in all the experiments, demonstrating that the fluorimetric microtitre assay has the necessary accuracy for high-throughput screening of new drugs against T. vaginalis.

Development of a simple, low cost chronoamperometric assay for fructose based on a commercial graphite-nanoparticle modified screen-printed carbon electrode.

Science.gov (United States)

Nicholas, Phil; Pittson, Robin; Hart, John P

2018-02-15

This paper describes the development of a simple, low cost chronoamperometric assay, for the measurement of fructose, using a graphite-nanoparticle modified screen-printed electrode (SPCE-G-COOH). Cyclic voltammetry showed that the response of the SPCE-G-COOH enhanced the sensitivity and precision, towards the enzymatically generated ferrocyanide species, over a plain SPCE; therefore the former was employed in subsequent studies. Calibration studies were carried out using chronoamperometry with a 40µl mixture containing fructose, mediator and FDH, deposited onto the SPCE-G-COOH. The response was linear from 0.1mM to 1.0mM. A commercial fruit juice sample was analysed using the developed assay and the fructose concentration was calculated to be 477mM with a precision of 3.03% (n=5). Following fortification (477mM fructose) the mean recovery was found to be 97.12% with a coefficient of variation of 6.42% (n=5); consequently, the method holds promise for the analysis of commercial fruit juices. Copyright © 2017 Elsevier Ltd. All rights reserved.

Quantitative analysis of Plasmodium ookinete motion in three dimensions suggests a critical role for cell shape in the biomechanics of malaria parasite gliding motility.

Science.gov (United States)

Kan, Andrey; Tan, Yan-Hong; Angrisano, Fiona; Hanssen, Eric; Rogers, Kelly L; Whitehead, Lachlan; Mollard, Vanessa P; Cozijnsen, Anton; Delves, Michael J; Crawford, Simon; Sinden, Robert E; McFadden, Geoffrey I; Leckie, Christopher; Bailey, James; Baum, Jake

2014-05-01

Motility is a fundamental part of cellular life and survival, including for Plasmodium parasites--single-celled protozoan pathogens responsible for human malaria. The motile life cycle forms achieve motility, called gliding, via the activity of an internal actomyosin motor. Although gliding is based on the well-studied system of actin and myosin, its core biomechanics are not completely understood. Currently accepted models suggest it results from a specifically organized cellular motor that produces a rearward directional force. When linked to surface-bound adhesins, this force is passaged to the cell posterior, propelling the parasite forwards. Gliding motility is observed in all three life cycle stages of Plasmodium: sporozoites, merozoites and ookinetes. However, it is only the ookinetes--formed inside the midgut of infected mosquitoes--that display continuous gliding without the necessity of host cell entry. This makes them ideal candidates for invasion-free biomechanical analysis. Here we apply a plate-based imaging approach to study ookinete motion in three-dimensional (3D) space to understand Plasmodium cell motility and how movement facilitates midgut colonization. Using single-cell tracking and numerical analysis of parasite motion in 3D, our analysis demonstrates that ookinetes move with a conserved left-handed helical trajectory. Investigation of cell morphology suggests this trajectory may be based on the ookinete subpellicular cytoskeleton, with complementary whole and subcellular electron microscopy showing that, like their motion paths, ookinetes share a conserved left-handed corkscrew shape and underlying twisted microtubular architecture. Through comparisons of 3D movement between wild-type ookinetes and a cytoskeleton-knockout mutant we demonstrate that perturbation of cell shape changes motion from helical to broadly linear. Therefore, while the precise linkages between cellular architecture and actomyosin motor organization remain unknown, our

Global optimization in the adaptive assay of subterranean uranium nodules

International Nuclear Information System (INIS)

Vulkan, U.; Ben-Haim, Y.

1989-01-01

An adaptive assay is one in which the design of the assay system is modified during operation in response to measurements obtained on-line. The present work has two aims: to design an adaptive system for borehole assay of isolated subterranean uranium nodules, and to investigate globality of optimal design in adaptive assay. It is shown experimentally that reasonably accurate estimates of uranium mass are obtained for a wide range of nodule shapes, on the basis of an adaptive assay system based on a simple geomorphological model. Furthermore, two concepts are identified which underlie the optimal design of the assay system. The adaptive assay approach shows promise for successful measurement of spatially random material in many geophysical applications. (author)

Chitosan scaffold modified with D-(+) raffinose and enriched with thiol-modified gelatin for improved osteoblast adhesion

International Nuclear Information System (INIS)

Galli, C; Parisi, L; Smerieri, A; Lumetti, S; Manfredi, E; Macaluso, G M; Elviri, L; Bianchera, A; Bettini, R; Lagonegro, P

2016-01-01

The aim of the present study was to investigate whether chitosan-based scaffolds modified with D-(+) raffinose and enriched with thiol-modified gelatin could selectively improve osteoblast adhesion and proliferation. 2, 3 and 4.5% chitosan films were prepared. Chitosan suitability for tissue engineering was confirmed by protein adsorption assay. Scaffolds were incubated with a 2.5 mg ml −1 BSA solution and the decrease of protein content in the supernatants was measured by spectrophotometry. Chitosan films were then enriched with thiol-modified gelatin and their ability to bind BSA was also measured. Then, 2% chitosan discs with or without thiol-modified gelatin were used as culture substrates for MC3T3-E1 cells. After 72 h cells were stained with trypan blue or with calcein AM and propidium iodide for morphology, viability and proliferation assays. Moreover, cell viability was measured at 48, 72, 96 and 168 h to obtain a growth curve. Chitosan films efficiently bound and retained BSA proportionally to the concentration of chitosan discs. The amount of protein retained was higher on chitosan enriched with thiol-modified gelatin. Moreover, chitosan discs allowed the adhesion and the viability of cells, but inhibited their proliferation. The functionalization of chitosan with thiol-modified gelatin enhanced cell spreading and proliferation. Our data confirm that chitosan is a suitable material for tissue engineering. Moreover, our data show that the enrichment of chitosan with thiol-modified gelatin enhances its biological properties. (paper)

Enzymatic assay for methotrexate in erythrocytes

DEFF Research Database (Denmark)

Schrøder, H; Heinsvig, E M

1985-01-01

Methotrexate (MTX) accumulates in erythrocytes in MTX-treated patients. We present a modified enzymatic assay measuring MTX concentrations between 10 and 60 nmol/l in erythrocytes, adapted for a centrifugal analyser (Cobas Bio). About 40 patient's samples could be analysed within 1 h. The detection...

Determination of hidden chromosome instability in persons suffered from the action of factors of the Chernobyl accident by the modified 'G2 bleomycin sensitivity assay'

International Nuclear Information System (INIS)

Dibs'kij, S.S.; Dibs'ka, O.B.; Pedan, L.P.; Pyilyins'ka, M.A.

2009-01-01

With the help of the modified 'G 2 -bleomycin sensitivity assay' the voluntary investigation of hidden chromosome instability in 53 persons with different radiation exposures had been fulfilled. In all examined groups, the individual levels of chromosome injuries under identical bleomycin exposure varied in a wide range and didn't depend on their initial values in intact cultures. Among control donors and individuals with low radiation exposure, ∼ 33 % hypertensive persons had been identified that can be considered as a genetically caused phenomenon. In patients recovered from acute radiation, 57.9 % persons expressed the hidden chromosome instability. The data obtained allow us to assume that high doses of ionizing radiation can modify the inherited susceptibility of human chromosomes to a mutagen exposure.

Determination of low tetanus or diphtheria antitoxin titers in sera by a toxin neutralization assay and a modified toxin-binding inhibition test

Directory of Open Access Journals (Sweden)

M.H. Sonobe

2007-01-01

Full Text Available A method for the screening of tetanus and diphtheria antibodies in serum using anatoxin (inactivated toxin instead of toxin was developed as an alternative to the in vivo toxin neutralization assay based on the toxin-binding inhibition test (TOBI test. In this study, the serum titers (values between 1.0 and 19.5 IU measured by a modified TOBI test (Modi-TOBI test and toxin neutralization assays were correlated (P < 0.0001. Titers of tetanus or diphtheria antibodies were evaluated in serum samples from guinea pigs immunized with tetanus toxoid, diphtheria-tetanus or triple vaccine. For the Modi-TOBI test, after blocking the microtiter plates, standard tetanus or diphtheria antitoxin and different concentrations of guinea pig sera were incubated with the respective anatoxin. Twelve hours later, these samples were transferred to a plate previously coated with tetanus or diphtheria antitoxin to bind the remaining anatoxin. The anatoxin was then detected using a peroxidase-labeled tetanus or diphtheria antitoxin. Serum titers were calculated using a linear regression plot of the results for the corresponding standard antitoxin. For the toxin neutralization assay, L+/10/50 doses of either toxin combined with different concentrations of serum samples were inoculated into mice for anti-tetanus detection, or in guinea pigs for anti-diphtheria detection. Both assays were suitable for determining wide ranges of antitoxin levels. The linear regression plots showed high correlation coefficients for tetanus (r² = 0.95, P < 0.0001 and for diphtheria (r² = 0.93, P < 0.0001 between the in vitro and the in vivo assays. The standardized method is appropriate for evaluating titers of neutralizing antibodies, thus permitting the in vitro control of serum antitoxin levels.

An ultra-sensitive colorimetric Hg(2+)-sensing assay based on DNAzyme-modified Au NP aggregation, MNPs and an endonuclease.

Science.gov (United States)

Li, Chao; Dai, Peiqing; Rao, Xinyi; Shao, Lin; Cheng, Guifang; He, Pingang; Fang, Yuzhi

2015-01-01

This paper reports the development of an ultra-sensitive colorimetric method for the detection of trace mercury ions involving DNAzymes, Au nanoparticle aggregation, magnetic nanoparticles and an endonuclease. DNAzyme-sensing elements are conjugated to the surface of Au nanoparticle-2, which can crosslink with the T-rich strands coated on Au nanoparticle-1 to form Au nanoparticle aggregation. Other T-rich stands are immobilized on the surface of MNPs. The specific hybridization of these two T-rich strands depends on the presence of Hg(2+), resulting in the formation of a T-Hg(2+)-T structure. Added endonuclease then digests the hybridized strands, and DNAzyme-modified Au NP aggregation is released, catalysing the conversion of the colourless ABTS into a blue-green product by H2O2-mediated oxidation. The increase in the adsorption spectrum of ABTS(+) at 421 nm is related to the concentration of Hg(2+). This assay was validated by detecting mercury ion concentrations in river water. The colorimetric responses were not significantly altered in the presence of 100-fold excesses of other metal ions such as Zn(2+), Pb(2+), Cd(2+), Mn(2+), Ca(2+) and Ni(2+). The inclusion of both Au NP aggregation and an endonuclease enables the assay to eliminate interference from the magnetic nanoparticles with colorimetric detection, decrease the background and improve the detection sensitivity. The calibration curve of the assay was linear over the range of Hg(2+) concentrations from 1 to 30 nM, and the detection limit was 0.8 nM, which is far lower than the 10 nM US EPA limit for drinking water. Copyright © 2014 Elsevier B.V. All rights reserved.

Complete genome sequence of the gliding, heparinolytic Pedobacter saltans type strain (113).

Science.gov (United States)

Liolios, Konstantinos; Sikorski, Johannes; Lu, Meagan; Nolan, Matt; Lapidus, Alla; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxanne; Han, Cliff; Goodwin, Lynne; Pitluck, Sam; Huntemann, Marcel; Ivanova, Natalia; Pagani, Ioanna; Mavromatis, Konstantinos; Ovchinikova, Galina; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam; Hauser, Loren; Brambilla, Evelyne-Marie; Kotsyurbenko, Oleg; Rohde, Manfred; Tindall, Brian J; Abt, Birte; Göker, Markus; Detter, John C; Woyke, Tanja; Bristow, James; Eisen, Jonathan A; Markowitz, Victor; Hugenholtz, Philip; Klenk, Hans-Peter; Kyrpides, Nikos C

2011-10-15

Pedobacter saltans Steyn et al. 1998 is one of currently 32 species in the genus Pedobacter within the family Sphingobacteriaceae. The species is of interest for its isolated location in the tree of life. Like other members of the genus P. saltans is heparinolytic. Cells of P. saltans show a peculiar gliding, dancing motility and can be distinguished from other Pedobacter strains by their ability to utilize glycerol and the inability to assimilate D-cellobiose. The genome presented here is only the second completed genome sequence of a type strain from a member of the family Sphingobacteriaceae to be published. The 4,635,236 bp long genome with its 3,854 protein-coding and 67 RNA genes consists of one chromosome, and is a part of the Genomic Encyclopedia of Bacteria and Archaea project.

Influence of the dislocation core on the glide of the 1/2 < 111 >{110} edge dislocation in bcc-iron: An embedded atom method study

Czech Academy of Sciences Publication Activity Database

Haghighat, S.M.H.; von Pezold, J.; Race, C. P.; Kormann, F.; Friák, Martin; Neugebauer, J.; Raabe, D.

2014-01-01

Roč. 87, MAY (2014), s. 274-282 ISSN 0927-0256 Institutional support: RVO:68081723 Keywords : Molecular dynamics * Edge dislocation * Core structure * Dislocation glide * Iron Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 2.131, year: 2014

Measurements of 3D slip velocities and plasma column lengths of a gliding arc discharge

DEFF Research Database (Denmark)

Zhu, Jiajian; Gao, Jinlong; Ehn, Andreas

2015-01-01

A non-thermal gliding arc discharge was generated at atmospheric pressure in an air flow. The dynamics of the plasma column and tracer particles were recorded using two synchronized high-speed cameras. Whereas the data analysis for such systems has previously been performed in 2D (analyzing......, gives more realistic insight into the convection cooling process. Additionally, with the determination of the 3D slip velocity and the 3D length of the plasma column, we give more accurate estimates for the drag force, the electric field strength, the power per unit length, and the radius...

Ribbon and gliding type parachutes evaluated in the 7 by 10 foot transonic wind tunnel

Energy Technology Data Exchange (ETDEWEB)

Ottensoser, J.

1975-09-01

An experiment has been conducted in the NSRDC 7- x 10-foot transonic tunnel for the Sandia Corporation to evaluate various parachute parameters. The experiment consisted of three main parts: the first phase evaluated the disreefing characteristics of the various parachutes as well as the drag forces before, during, and after disreefing; the second phase measured the pressure distribution around the chute as well as the drag forces; and the final phase evaluated the disreefing and drag characteristics of gliding type parachutes. The free stream dynamic pressure varied from 65 to 500 psf. 12 figures, 1 table. (auth)

Radioenzymatic assay of DOPA (3,4-dihydroxyphenylalanine)

International Nuclear Information System (INIS)

Johnson, G.A.; Gren, J.M.; Kupiecki, R.

1978-01-01

We modified the single-isotope radioenzymatic assay for catecholamines [Life Sci. 21, 625(1977)] to assay 3,4-dihydroxyphenylalanine (DOPA). DOPA decarboxylase is used to convert DOPA to dopamine, which concurrently is converted to [ 3 H]-3-O-methyldopamine in the presence of catechol-O-methyltransferase and [methyl- 3 H]-S-adenosylmethionine and assayed radioenzymatically. For assay of plasma DOPA, 50 μl of untreated plasma is added directly into the incubation mixture. A duplicate mixture containing an internal standard requires a second 50-μl aliquot of plasma. Because the assay measures both DOPA and endogenous dopamine, two additional aliquots of plasma must be assayed for dopamine in the absence of the decarboxylase by the differential assay; DOPA is estimated by difference. The assay is sensitive to 25 pg (500 ng/liter of plasma). Analysis of DOPA (DOPA plus dopamine) and the concurrent differential assay of catecholamines in at least 10 samples can be done in a single working day. Plasma DOPA concentrations for 42 normotensive adults were 1430 +- 19 ng/liter (mean +- SEM). In contrast, dopamine concentrations for these same subjects averaged 23 +- 20 ng/liter. Values for the 24 women subjects (1510 +- 62 ng/liter) significantly (P = 0.04) exceeded those for the men

Development of fluorescent methods for DNA methyltransferase assay

Science.gov (United States)

Li, Yueying; Zou, Xiaoran; Ma, Fei; Tang, Bo; Zhang, Chun-yang

2017-03-01

DNA methylation modified by DNA methyltransferase (MTase) plays an important role in regulating gene transcription, cell growth and proliferation. The aberrant DNA MTase activity may lead to a variety of human diseases including cancers. Therefore, accurate and sensitive detection of DNA MTase activity is crucial to biomedical research, clinical diagnostics and therapy. However, conventional DNA MTase assays often suffer from labor-intensive operations and time-consuming procedures. Alternatively, fluorescent methods have significant advantages of simplicity and high sensitivity, and have been widely applied for DNA MTase assay. In this review, we summarize the recent advances in the development of fluorescent methods for DNA MTase assay. These emerging methods include amplification-free and the amplification-assisted assays. Moreover, we discuss the challenges and future directions of this area.

Improved MPSP Method-based Cooperative Re-entry Guidance for Hypersonic Gliding Vehicles

Directory of Open Access Journals (Sweden)

Chu Haiyan

2017-01-01

Full Text Available A computationally sufficient technique is used to solve the 3-D cooperative re-entry guidance problem for hypersonic gliding vehicles. Due to the poor surrounding adaptive ability of the traditional cooperative guidance methods, a novel methodology, named as model predictive static programming (MPSP, is used to solve a class of finite-horizon optimal control problems with hard terminal constraints. The main feature of this guidance law is that it is capable of hitting the target with high accuracy for each one of the cooperative vehicles at the same time. In addition, it accurately satisfies variable constraints. Performance of the proposed MPSP-based guidance is demonstrated in 3-D nonlinear dynamics scenario. The numerical simulation results show that the proposed cooperative re-entry guidance methodology has the advantage of computational efficiency and better robustness against the perturbations.

Automated DNA extraction from genetically modified maize using aminosilane-modified bacterial magnetic particles.

Science.gov (United States)

Ota, Hiroyuki; Lim, Tae-Kyu; Tanaka, Tsuyoshi; Yoshino, Tomoko; Harada, Manabu; Matsunaga, Tadashi

2006-09-18

A novel, automated system, PNE-1080, equipped with eight automated pestle units and a spectrophotometer was developed for genomic DNA extraction from maize using aminosilane-modified bacterial magnetic particles (BMPs). The use of aminosilane-modified BMPs allowed highly accurate DNA recovery. The (A(260)-A(320)):(A(280)-A(320)) ratio of the extracted DNA was 1.9+/-0.1. The DNA quality was sufficiently pure for PCR analysis. The PNE-1080 offered rapid assay completion (30 min) with high accuracy. Furthermore, the results of real-time PCR confirmed that our proposed method permitted the accurate determination of genetically modified DNA composition and correlated well with results obtained by conventional cetyltrimethylammonium bromide (CTAB)-based methods.

Ceratoplastia endotelial com desnudamento da Descemet(DSEK utilizando o dispositivo TAN EndoGlideTM: série de casos

Directory of Open Access Journals (Sweden)

Henrique Santiago Baltar Pazos

2011-06-01

Full Text Available OBJETIVO: Relatar os resultados da ceratoplastia endotelial com desnudamento da Descemet (DSEK utilizando o dispositivo TAN EndoGlideTM para facilitar a introdução da membrana endotelial. MÉTODOS: Série de casos consecutivos, prospectiva. Foram incluídos 9 pacientes com edema corneano secundário à disfunção endotelial. Melhor acuidade visual corrigida, refração, astigmatismo ceratométrico, espessura corneana central, densidade das células endoteliais e complicações foram analisadas após seguimento de seis meses. RESULTADOS: Houve melhora do edema de córnea e da visão em 7 pacientes (77,78%. A melhor acuidade visual corrigida ficou entre 20/40 e 20/200. A densidade endotelial média após 6 meses variou entre 1.305 céls/mm² e 2.346 céls/mm² com média de perda de 33,14%. Desprendimento de parte do enxerto ocorreu em 1 olho (11,11%, falência primária do transplante endotelial em 2 olhos (22,22%. CONCLUSÃO: O dispositivo TAN EndoGlideTM facilita a introdução do enxerto na ceratoplastia endotelial com desnudamento da Descemet.

Development, Optimization, and Evaluation of a Duplex Droplet Digital PCR Assay To Quantify the T-nos/hmg Copy Number Ratio in Genetically Modified Maize.

Science.gov (United States)

Félix-Urquídez, Dalmira; Pérez-Urquiza, Melina; Valdez Torres, José-Benigno; León-Félix, Josefina; García-Estrada, Raymundo; Acatzi-Silva, Abraham

2016-01-05

Certified reference materials (CRMs) are required to guarantee the reliability of analytical measurements. The CRMs available in the field of genetically modified organisms (GMOs) are characterized using real-time polymerase chain reaction (qPCR). This technology has limited application, because of its dependence on a calibrant. The objective of this study was to obtain a method with higher metrological quality, to characterize the CRMs for their contents of T-nos/hmg copy number ratio in maize. A duplex droplet digital PCR (ddPCR) assay was developed and optimized by a central composite design. The developed method achieved an absolute limit of detection (LOD) of 11 cP T-nos, a relative LOD of 0.034%, a limit of quantification (LOQ) of 23 cP (relative LOQ of 0.08%), and a dynamic range of 0.08%-100% T-nos/hmg ratio. The specificity and applicability of the assay were established for the analysis of low T-nos concentrations (0.9%) in several corn varieties. The convenience of DNA digestion to reduce measurement bias in the case of multiple-copy binding was confirmed through an enzymatic restriction assay. Given its overall performance, this method can be used to characterize CRM candidates for their contents of T-nos/hmg ratio.

Development, Optimization, and Evaluation of a Duplex Droplet Digital PCR Assay To Quantify the T-nos/hmg Copy Number Ratio in Genetically Modified Maize

Science.gov (United States)

2015-01-01

Certified reference materials (CRMs) are required to guarantee the reliability of analytical measurements. The CRMs available in the field of genetically modified organisms (GMOs) are characterized using real-time polymerase chain reaction (qPCR). This technology has limited application, because of its dependence on a calibrant. The objective of this study was to obtain a method with higher metrological quality, to characterize the CRMs for their contents of T-nos/hmg copy number ratio in maize. A duplex droplet digital PCR (ddPCR) assay was developed and optimized by a central composite design. The developed method achieved an absolute limit of detection (LOD) of 11 cP T-nos, a relative LOD of 0.034%, a limit of quantification (LOQ) of 23 cP (relative LOQ of 0.08%), and a dynamic range of 0.08%–100% T-nos/hmg ratio. The specificity and applicability of the assay were established for the analysis of low T-nos concentrations (0.9%) in several corn varieties. The convenience of DNA digestion to reduce measurement bias in the case of multiple-copy binding was confirmed through an enzymatic restriction assay. Given its overall performance, this method can be used to characterize CRM candidates for their contents of T-nos/hmg ratio. PMID:26605751

In situ TEM study on elastic interaction between a prismatic loop and a gliding dislocation

International Nuclear Information System (INIS)

Matsukawa, Yoshitaka; Liu, Grace S.

2012-01-01

In situ straining in a transmission electron microscope was performed in order to investigate dislocation interactions with a prismatic loop, which as a mobile obstacle is expected to be displaced by the strain-field of dislocation prior to physical contact. It was found that when a gliding dislocation approached a critical distance, the prismatic loop was certainly attracted to the dislocation. The captured loop disrupted the dislocation motion and was not dragged along with the mobile dislocation. Instead, the dislocation bypassed the loop via cross-slip to another slip plane with a resolved shear stress estimated to be 40% lower than that of the original plane.

A modified direct insulin lodination for insulin radioreceptor assay in human erythrocytes

Energy Technology Data Exchange (ETDEWEB)

Elnabrawie, F S; Megahed, Y M; Fahim, F A; Ahmed, A M [Middle Eastern Regional radioisotope center for Arab Countries biochemistry dept. Faculty of science Ain Shams university, cairo, (Egypt)

1995-10-01

A substitution of iodine in to the inulin molecule will easily lead to a decreased hormonal activity. Nevertheless it cannot be concluded without further investigation that one or several of the tyrosyl groups are directly involve in the degree of iodination and the loss activity has been demonstrated by many workers, (Frenkel-Conrat 1950, and de Zoeten and van Strik, 1961). On the basis of the Frenkel-Conrat experiments, lee 1957 suggested that a mono-substitution of iodine in two tyrosyl groups is possible without loss of biological activity. In general, iodination is easy to perform as usually carried out at room temperature at PH of 7.5 in a phosphate buffer medium (Megahed et al., 1976, 1979). Reaction between antigen and antibody in radioimmunoassay (RIA), immunoradiometric assay (IRMA) and radioreceptor assay (RRA), is detected by radiation emitted from a radioisotope incorporated into the antigen or antibody molecule. Ideally the radiolabelled molecule has an immunoreactivity identical to the natural molecule and this behaves in the same way as in the assay procedure. Oxidation of 125 I-iodide gives rise to the 125 I-iodination which in a mildly alkaline PH (7.5) reacts with the phenolic benzene ring of tyrosine and tyrosyl residues by a process of electrophilic attack.

A modified direct insulin lodination for insulin radioreceptor assay in human erythrocytes

International Nuclear Information System (INIS)

Elnabrawie, F.S.; Megahed, Y.M.; Fahim, F.A.; Ahmed, A.M.

1995-01-01

A substitution of iodine in to the inulin molecule will easily lead to a decreased hormonal activity. Nevertheless it cannot be concluded without further investigation that one or several of the tyrosyl groups are directly involve in the degree of iodination and the loss activity has been demonstrated by many workers, (Frenkel-Conrat 1950, and de Zoeten and van Strik, 1961). On the basis of the Frenkel-Conrat experiments, lee 1957 suggested that a mono-substitution of iodine in two tyrosyl groups is possible without loss of biological activity. In general, iodination is easy to perform as usually carried out at room temperature at PH of 7.5 in a phosphate buffer medium (Megahed et al., 1976, 1979). Reaction between antigen and antibody in radioimmunoassay (RIA), immunoradiometric assay (IRMA) and radioreceptor assay (RRA), is detected by radiation emitted from a radioisotope incorporated into the antigen or antibody molecule. Ideally the radiolabelled molecule has an immunoreactivity identical to the natural molecule and this behaves in the same way as in the assay procedure. Oxidation of 125 I-iodide gives rise to the 125 I-iodination which in a mildly alkaline PH (7.5) reacts with the phenolic benzene ring of tyrosine and tyrosyl residues by a process of electrophilic attack

An ARGS-aggrecan assay for analysis in blood and synovial fluid

DEFF Research Database (Denmark)

Larsson, S; Lohmander, Stefan; Struglics, A

2014-01-01

OBJECTIVE: To validate a modified ligand-binding assay for the detection of aggrecanase generated aggrecan fragments with the ARGS neoepitope in synovial fluid (SF) and blood, and to verify the identity of aggrecan fragments found in blood. DESIGN: An enzyme-linked immunosorbent assay (ELISA....... Aggrecan was purified from serum and plasma pools and analysed by Western blot. RESULTS: The limits of quantification for the ARGS-aggrecan assay was between 0.2 and 0.025 pmol ARGS/ml, and the sensitivity of the assay was improved two-fold compared to when using a standard purified from human donors...... similar, and correlated (r(S) = 0.773, P assay is highly sensitive and suited for analysis...

The electrophoretic mobility shift assay (EMSA)

OpenAIRE

sprotocols

2015-01-01

The electrophoretic mobility shift assay (EMSA), also known as “gel shift assay”, is used to examine the binding parameters and relative affinities of protein and DNA interactions. We produced recombinant CCA1 protein and tested its binding affinity for the promoter fragments that contain CBS (AAAAATCT) or evening element (EE, AAAATATCT) (1) using a modified procedure adopted from published protocols (2,3).

Cholinesterase assay by an efficient fixed time endpoint method

Directory of Open Access Journals (Sweden)

Mónica Benabent

2014-01-01

The method may be adapted to the user needs by modifying the enzyme concentration and applied for simultaneously testing many samples in parallel; i.e. for complex experiments of kinetics assays with organophosphate inhibitors in different tissues.

A novel hematoxylin and eosin stain assay for detection of the parasitic dinoflagellate Amoebophrya.

Science.gov (United States)

Li, Caiwen; Chen, Tiantian

2017-02-01

The parasitic dinoflagellate Amoebophrya infects broad range of marine organisms. Particularly, Amoebophrya infections in planktonic dinoflagellates can prevent or delay the formation of algal blooms, and recycle undergrazed planktonic dinoflagellates back to the microbial loop by disrupting host cells. Its ecological significance was gradually recognized along with the discovery of its enormous molecular diversity in oceanic and coastal ecosystems. Thus, we developed a reliable, easily accessible and less time-consuming assay, to detect and assess Amoebophrya infections in planktonic dinoflagellates. The modified hematoxylin and eosin staining assay provided reliable diagnosis of Amoebophrya infection by identifying the characteristic "beehive" of the multinucleate trophonts. After staining, the typical multinucleate "beehive" is evidently distinguishable from the compact nuclei of uninfected host cells. The modified hematoxylin and eosin (H & E) staining assay is easy to use, that can be routinely performed within 3h (up to 20 samples/batch) using general laboratory equipment, supplies and chemical reagents. The produced slides with agar-embedded dinoflagellate cells can be stored for several months or even years in a dry place without noticeable loss in quality of staining. With suitable calculation, the modified H & E assay can be applied to assess the prevalence of Amoebophrya infection in planktonic dinoflagellates. This efficient and powerful assay will facilitate the investigation on the ecological roles of Amoebophryidae in coastal and oceanic ecosystem. Copyright © 2016 Elsevier B.V. All rights reserved.

Comparison of tetrazolium colorimetric and [3H]-uridine assays for in vitro chemosensitivity testing.

Science.gov (United States)

Ford, C H; Richardson, V J; Tsaltas, G

1989-01-01

We have routinely used a [3H]-uridine microplate assay for assessing chemosensitivity. A colorimetric assay with the advantages of safety, cost and simplicity has previously been described and relies on the ability of living cells to reduce a soluble tetrazolium salt, 3-4,5-dimethylthiazol-2,5-diphenyl-tetrazolium bromide (MMT), into an insoluble formazan precipitate. We compared the chemosensitivity of 14 human tumour cell lines of colonic, lung and cervical carcinoma origin to doxorubicin, vindesine or vindesine immunoconjugates in both the [3H]-uridine assay and a modified MTT assay to evaluate whether we could change to the non-radiolabelled method. Correlation between the concentration of drug causing 50% inhibition of cell growth (IC50) for these agents between the two assays was very poor. However, taking account of recent reports in the literature, we modified the MTT assay by removing serum-containing medium and using dimethyl sulphoxide to solubilise the formazan precipitate. This considerably improved the correlation between the assays for doxorubicin (r = 0.871; P = 0.001) and vindesine (r = 0.981; P less than 0.001). Our data indicates that the MTT assay can be used to replace the [3H]-uridine assay for chemosensitivity screening, but further modifications are necessary to improve the sensitivity and decrease the problem of cell loss after washing, which was noted with some adherent cell lines.

Complete genome sequence of the gliding, heparinolytic Pedobacter saltans type strain (113T)

Science.gov (United States)

Liolios, Konstantinos; Sikorski, Johannes; Lu, Meagan; Nolan, Matt; Lapidus, Alla; Lucas, Susan; Hammon, Nancy; Deshpande, Shweta; Cheng, Jan-Fang; Tapia, Roxanne; Han, Cliff; Goodwin, Lynne; Pitluck, Sam; Huntemann, Marcel; Ivanova, Natalia; Pagani, Ioanna; Mavromatis, Konstantinos; Ovchinikova, Galina; Pati, Amrita; Chen, Amy; Palaniappan, Krishna; Land, Miriam; Hauser, Loren; Brambilla, Evelyne-Marie; Kotsyurbenko, Oleg; Rohde, Manfred; Tindall, Brian J.; Abt, Birte; Göker, Markus; Detter, John C.; Woyke, Tanja; Bristow, James; Eisen, Jonathan A.; Markowitz, Victor; Hugenholtz, Philip; Klenk, Hans-Peter; Kyrpides, Nikos C.

2011-01-01

Pedobacter saltans Steyn et al. 1998 is one of currently 32 species in the genus Pedobacter within the family Sphingobacteriaceae. The species is of interest for its isolated location in the tree of life. Like other members of the genus P. saltans is heparinolytic. Cells of P. saltans show a peculiar gliding, dancing motility and can be distinguished from other Pedobacter strains by their ability to utilize glycerol and the inability to assimilate D-cellobiose. The genome presented here is only the second completed genome sequence of a type strain from a member of the family Sphingobacteriaceae to be published. The 4,635,236 bp long genome with its 3,854 protein-coding and 67 RNA genes consists of one chromosome, and is a part of the Genomic Encyclopedia of Bacteria and Archaea project. PMID:22180808

Clonogenic assay: adherent cells.

Science.gov (United States)

Rafehi, Haloom; Orlowski, Christian; Georgiadis, George T; Ververis, Katherine; El-Osta, Assam; Karagiannis, Tom C

2011-03-13

The clonogenic (or colony forming) assay has been established for more than 50 years; the original paper describing the technique was published in 1956. Apart from documenting the method, the initial landmark study generated the first radiation-dose response curve for X-ray irradiated mammalian (HeLa) cells in culture. Basically, the clonogenic assay enables an assessment of the differences in reproductive viability (capacity of cells to produce progeny; i.e. a single cell to form a colony of 50 or more cells) between control untreated cells and cells that have undergone various treatments such as exposure to ionising radiation, various chemical compounds (e.g. cytotoxic agents) or in other cases genetic manipulation. The assay has become the most widely accepted technique in radiation biology and has been widely used for evaluating the radiation sensitivity of different cell lines. Further, the clonogenic assay is commonly used for monitoring the efficacy of radiation modifying compounds and for determining the effects of cytotoxic agents and other anti-cancer therapeutics on colony forming ability, in different cell lines. A typical clonogenic survival experiment using adherent cells lines involves three distinct components, 1) treatment of the cell monolayer in tissue culture flasks, 2) preparation of single cell suspensions and plating an appropriate number of cells in petri dishes and 3) fixing and staining colonies following a relevant incubation period, which could range from 1-3 weeks, depending on the cell line. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cell lines with the use of an immortalized human keratinocyte cell line (FEP-1811). Also, our aims are to describe common features of clonogenic assays including calculation of the plating efficiency and survival fractions after exposure of cells to radiation, and to exemplify modification of radiation-response with the use of a natural antioxidant

Comparison of Modified Impedance Whole Blood Platelet Aggregation Method Detecting Platelet Function in ACS Patients with Different CYP2C19 Genotypes.

Science.gov (United States)

Cui, Chanjuan; Qiao, Rui; Zhang, Jie

2016-01-01

A reliable laboratory test to monitor onclopidogrel platelet reactivity (PR) is very necessary. In addition, genetic factors also play an important part in onclopidogrel PR. This study aimed to modify the original impedance whole blood platelet aggregation assay associated with the release assay to monitor onclopidogrel PR and assess their relationship with genotype. We adjusted the concentration of calcium in the in vitro reaction system of platelet aggregation to modify the original impedance whole blood platelet aggregation assay. Meanwhile, chronolume, which quantified the adenosine triphosphate (ATP) released from platelet dense granules, is added to this reaction system to reflect the platelet release function. In the modified assay, platelet magnified activation time (MAT) and the maximal platelet ATP release value (RV) were used to reflect platelet function parameters. In the original assay, the electrical resistance (omega) and RV were used to reflect platelet function parameters. Onclopidogrel PR was detected by the original impedance whole blood platelet aggregation assay, modified assay, and flow cytometric vasodilator stimulated phosphoprotein (VASP) assay in 168 patients with acute coronary syndromes (ACS). CYP2C19*2 and CYP2C19*3 polymorphisms were also detected in all of these patients. This modified method showed that when 12.5 microL CaCl2 (0.2 mmol/L) was added to the reaction system, MAT was appropriate (93 +/- 23 seconds). The CVs for the modified impedance assay and release assay were 9.31% and 6.13%, respectively. The mean VASP-PRI in the patient group treated with clopidogrel was significantly lower than that in the control group without antiplatelet therapy (54.88 +/- 16.81% vs. 79.86 +/- 10.24%, p 50% group were shorter than that in the PRI 50% group were higher than that in the PRI omega) and RV of the original method showed no differences between the two groups [0 (0-2) vs. 0 (0-1.25), 0.05 (0-0.25) vs. 0.08 (0-0.24); p > 0.05, p > 0

Treatment by gliding arc of epoxy resin: preliminary analysis of surface modifications

Science.gov (United States)

Faubert, F.; Wartel, M.; Pellerin, N.; Pellerin, S.; Cochet, V.; Regnier, E.; Hnatiuc, B.

2016-12-01

Treatments with atmospheric pressure non-thermal plasma are easy to implement and inexpensive. Among them gliding arc (GlidArc) remains rarely used in surface treatment of polymers. However, it offers economic and flexible way to treat quickly large areas. In addition the choice of carrier gas makes it possible to bring the active species and other radicals allowing different types of grafting and functionalization of the treated surfaces, for example in order to apply for anti-biofouling prevention. This preliminary work includes analysis of the surface of epoxy resins by infrared spectroscopy: the different affected chemical bonds were studied depending on the duration of treatment. The degree of oxidation (the C/O ratio) is obtained by X-ray microanalysis and contact angle analysis have been performed to determinate the wettability properties of the treated surface. A spectroscopic study of the plasma allows to determine the possible active species in the different zones of the discharge.

Radioreceptor assay analysis of tamsulosin and terazosin pharmacokinetics

NARCIS (Netherlands)

Taguchi, K.; Schäfers, R. F.; Michel, M. C.

1998-01-01

AIMS: A radioreceptor assay has been developed for alpha1-adrenoceptor subtypes and applied to a pharmacokinetic analysis of tamsulosin and terazosin. METHODS: Young, male, healthy volunteers received 0.4 mg tamsulosin (as Omnic modified release capsules) or 5 mg terazosin (as Flotrin tablets) in a

[Effectiveness of the GlideScope video laryngoscope in a case of unexpected difficult airway due to lingual tonsil hypertrophy].

Science.gov (United States)

Cruz, P; Alarcón, L; Del Castillo, T; Cabrerizo, P; Díaz, S

2015-05-01

Lingual tonsil hypertrophy can cause varying degrees of airway obstruction and is considered a risk factor for difficult mask ventilation and tracheal intubation. We report a case of unexpected difficult airway in a patient with unknown lingual tonsil hypertrophy that was solved with the use of the GlideScope video laryngoscope. Copyright © 2014 Sociedad Española de Anestesiología, Reanimación y Terapéutica del Dolor. Publicado por Elsevier España, S.L.U. All rights reserved.

Further characterization of benzo[a]pyrene diol-epoxide (BPDE)-induced comet assay effects.

Science.gov (United States)

Bausinger, Julia; Schütz, Petra; Piberger, Ann Liza; Speit, Günter

2016-03-01

The present study aims to further characterize benzo[a]pyrene diol-epoxide (BPDE)-induced comet assay effects. Therefore, we measured DNA effects by the comet assay and adduct levels by high-performance liquid chromatography (HPLC) in human lymphocytes and A549 cells exposed to (±)-anti-benzo[a]pyrene-7,8-diol 9,10-epoxide [(±)-anti-BPDE] or (+)-anti-benzo[a]pyrene-7,8-diol 9,10-epoxide [(+)-anti-BPDE]. Both, the racemic form and (+)-anti-BPDE, which is the most relevant metabolite with regard to mutagenicity and carcinogenicity, induced DNA migration in cultured lymphocytes in the same range of concentrations to a similar extent in the alkaline comet assay after exposure for 2h. Nevertheless, (+)-anti-BPDE induced significantly enhanced DNA migration after 16 and 18h post-cultivation which was not seen in response to (±)-anti-BPDE. Combination of the comet assay with the Fpg (formamidopyrimidine-DNA glycosylase) protein did not enhance BPDE-induced effects and thus indicated the absence of Fpg-sensitive sites (oxidized purines, N7-guanine adducts, AP-sites). The aphidicolin (APC)-modified comet assay suggested significant excision repair activity of cultured lymphocytes during the first 18h of culture after a 2 h-exposure to BPDE. In contrast to these repair-related effects measured by the comet assay, HPLC analysis of stable adducts did not reveal any significant removal of (+)-anti-BPDE-induced adducts from lymphocytes during the first 22h of culture. On the other hand, HPLC measurements indicated that A549 cells repaired about 70% of (+)-anti-BPDE-induced DNA-adducts within 22h of release. However, various experiments with the APC-modified comet assay did not indicate significant repair activity during this period in A549 cells. The conflicting results obtained with the comet assay and the HPLC-based adduct analysis question the real cause for BPDE-induced DNA migration in the comet assay and the reliability of the APC-modified comet assay for the

The comet assay: ready for 30 more years.

Science.gov (United States)

Møller, Peter

2018-02-24

During the last 30 years, the comet assay has become widely used for the measurement of DNA damage and repair in cells and tissues. A landmark achievement was reached in 2016 when the Organization for Economic Co-operation and Development adopted a comet assay guideline for in vivo testing of DNA strand breaks in animals. However, the comet assay has much more to offer than being an assay for testing DNA strand breaks in animal organs. The use of repair enzymes increases the range of DNA lesions that can be detected with the assay. It can also be modified to measure DNA repair activity. Still, despite the long-term use of the assay, there is a need for studies that assess the impact of variation in specific steps of the procedure. This is particularly important for the on-going efforts to decrease the variation between experiments and laboratories. The articles in this Special Issue of Mutagenesis cover important technical issues of the comet assay procedure, nanogenotoxicity and ionising radiation sensitivity on plant cells. The included biomonitoring studies have assessed seasonal variation and certain predictors for the basal level of DNA damage in white blood cells. Lastly, the comet assay has been used in studies on genotoxicity of environmental and occupational exposures in human biomonitoring studies and animal models. Overall, the articles in this Special Issue demonstrate the versatility of the comet assay and they hold promise that the assay is ready for the next 30 years.

Comparison of static and microfluidic protease assays using modified bioluminescence resonance energy transfer chemistry.

Directory of Open Access Journals (Sweden)

Nan Wu

Full Text Available BACKGROUND: Fluorescence and bioluminescence resonance energy transfer (F/BRET are two forms of Förster resonance energy transfer, which can be used for optical transduction of biosensors. BRET has several advantages over fluorescence-based technologies because it does not require an external light source. There would be benefits in combining BRET transduction with microfluidics but the low luminance of BRET has made this challenging until now. METHODOLOGY: We used a thrombin bioprobe based on a form of BRET (BRET(H, which uses the BRET(1 substrate, native coelenterazine, with the typical BRET(2 donor and acceptor proteins linked by a thrombin target peptide. The microfluidic assay was carried out in a Y-shaped microfluidic network. The dependence of the BRET(H ratio on the measurement location, flow rate and bioprobe concentration was quantified. Results were compared with the same bioprobe in a static microwell plate assay. PRINCIPAL FINDINGS: The BRET(H thrombin bioprobe has a lower limit of detection (LOD than previously reported for the equivalent BRET(1-based version but it is substantially brighter than the BRET(2 version. The normalised BRET(H ratio of the bioprobe changed 32% following complete cleavage by thrombin and 31% in the microfluidic format. The LOD for thrombin in the microfluidic format was 27 pM, compared with an LOD of 310 pM, using the same bioprobe in a static microwell assay, and two orders of magnitude lower than reported for other microfluidic chip-based protease assays. CONCLUSIONS: These data demonstrate that BRET based microfluidic assays are feasible and that BRET(H provides a useful test bed for optimising BRET-based microfluidics. This approach may be convenient for a wide range of applications requiring sensitive detection and/or quantification of chemical or biological analytes.

The development and validation of EpiComet-Chip, a modified high-throughput comet assay for the assessment of DNA methylation status.

Science.gov (United States)

Townsend, Todd A; Parrish, Marcus C; Engelward, Bevin P; Manjanatha, Mugimane G

2017-08-01

DNA damage and alterations in global DNA methylation status are associated with multiple human diseases and are frequently correlated with clinically relevant information. Therefore, assessing DNA damage and epigenetic modifications, including DNA methylation, is critical for predicting human exposure risk of pharmacological and biological agents. We previously developed a higher-throughput platform for the single cell gel electrophoresis (comet) assay, CometChip, to assess DNA damage and genotoxic potential. Here, we utilized the methylation-dependent endonuclease, McrBC, to develop a modified alkaline comet assay, "EpiComet," which allows single platform evaluation of genotoxicity and global DNA methylation [5-methylcytosine (5-mC)] status of single-cell populations under user-defined conditions. Further, we leveraged the CometChip platform to create an EpiComet-Chip system capable of performing quantification across simultaneous exposure protocols to enable unprecedented speed and simplicity. This system detected global methylation alterations in response to exposures which included chemotherapeutic and environmental agents. Using EpiComet-Chip on 63 matched samples, we correctly identified single-sample hypermethylation (≥1.5-fold) at 87% (20/23), hypomethylation (≥1.25-fold) at 100% (9/9), with a 4% (2/54) false-negative rate (FNR), and 10% (4/40) false-positive rate (FPR). Using a more stringent threshold to define hypermethylation (≥1.75-fold) allowed us to correctly identify 94% of hypermethylation (17/18), but increased our FPR to 16% (7/45). The successful application of this novel technology will aid hazard identification and risk characterization of FDA-regulated products, while providing utility for investigating epigenetic modes of action of agents in target organs, as the assay is amenable to cultured cells or nucleated cells from any tissue. Environ. Mol. Mutagen. 58:508-521, 2017. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis.

Science.gov (United States)

Khairnar, Krishna; Sanmukh, Swapnil; Chandekar, Rajshree; Paunikar, Waman

2014-07-01

The comet assay is the widely used method for in vitro toxicity testing which is also an alternative to the use of animal models for in vivo testing. Since, its inception in 1984 by Ostling and Johansson, it is being modified frequently for a wide range of application. In spite of its wide applicability, unfortunately there is no report of its application in bacteriophages research. In this study, a novel application of comet assay for the detection of bacteriophage mediated bacterial cell lysis was described. The conventional methods in bacteriophage research for studying bacterial lysis by bacteriophages are plaque assay method. It is time consuming, laborious and costly. The lytic activity of bacteriophage devours the bacterial cell which results in the release of bacterial genomic material that gets detected by ethidium bromide staining method by the comet assay protocol. The objective of this study was to compare efficacy of comet assay with different assay used to study phage mediated bacterial lysis. The assay was performed on culture isolates (N=80 studies), modified comet assay appear to have relatively higher sensitivity and specificity than other assay. The results of the study showed that the application of comet assay can be an economical, time saving and less laborious alternative to conventional plaque assay for the detection of bacteriophage mediated bacterial cell lysis. Copyright © 2014 Elsevier B.V. All rights reserved.

Atomic-scale nanoindentation: detection and identification of single glide events in three dimensions by force microscopy

International Nuclear Information System (INIS)

Egberts, P; Bennewitz, R

2011-01-01

Indentation experiments on the nanometre scale have been performed by means of atomic force microscopy in ultra-high vacuum on KBr(100) surfaces. The surfaces yield in the form of discrete surface displacements with a typical length scale of 1 A. These surface displacements are detected in both normal and lateral directions. Measurement of the lateral tip displacement requires a load-dependent calibration due to the load dependence of the effective lateral compliance. Correlation of the lateral and normal displacements for each glide event allow identification of the activated slip system. The results are discussed in terms of the resolved shear stress in indentation experiments and of typical results in atomistic simulations of nanometre-scale indentation.

Leaping shampoo glides on a 500-nm-thick lubricating air layer

Science.gov (United States)

Li, Erqiang; Lee, Sanghyun; Marston, Jeremy; Bonito, Andrea; Thoroddsen, Sigurdur

2013-11-01

When a stream of shampoo is fed onto a pool in one's hand, a jet can leap sideways or rebound from the liquid surface in an intriguing phenomenon known as the Kaye effect. Earlier studies have debated whether non-Newtonian effects are the underlying cause of this phenomenon, making the jet glide on top of a shear-thinning liquid layer, or whether an entrained air layer is responsible. Herein we show unambiguously that the jet slides on a lubricating air layer [Lee et al., Phys. Rev. E 87, 061001 (2013)]. We identify this layer by looking through the pool liquid and observing its rupture into fine micro-bubbles. The resulting micro-bubble sizes suggest that the thickness of this air layer is around 500 nm. This thickness estimate is also supported by the tangential deceleration of the jet during the rebounding, with the shear stress within the thin air layer sufficient for the observed deceleration. Particle tracking within the jet shows uniform velocity, with no pronounced shear, which would be required for shear-thinning effects. The role of the surfactant may primarily be to stabilize the air film.

In vitro comparison of the cyclic fatigue resistance of HyFlex EDM, One G, and ProGlider nickel titanium glide path instruments in single and double curvature canals.

Science.gov (United States)

Yılmaz, Koray; Uslu, Gülşah; Özyürek, Taha

2017-11-01

It was aimed to compare the cyclic fatigue resistances of ProGlider (PG), One G (OG), and HyFlex EDM (HEDM) nickel titanium glide path files in single- and double-curved artificial canals. 40 PG (16/0.02), 40 OG (14/0.03), and 40 HEDM (10/0.05) single-file glide path files were used in the present study. Sixty files were subjected to cyclic fatigue test by using double-curved canals and 60 files by using single-curved canal ( n = 20). The number of cycles to fracture (NCF) was calculated and the length of the fractured fragment (FL) was determined by a digital micro-caliper. Twelve pieces of fractured files were examined with scanning electron microscope to determine fracture types of the files ( n = 2). The NCF and the FL data were analyzed using one-way analysis of variance and post hoc Tukey test using SPSS 21 software ( p < 0.05). In all of the groups, NCF values were significantly lower in double-curved canals when compared to single-curved canals ( p < 0.05). For both of single- and double-curved canals, NCF values of HEDM group in apical and coronal curvatures were found to be significantly higher than NCF values of PG and OG groups ( p < 0.05). In both of single- and double-curved canals, NCF value of PG group was found significantly higher than OG group ( p < 0.05). Within the limitations of this study, HEDM glide path files were found to have the highest cyclic fatigue resistance in both of single- and double-curved canals.

A dual amplification strategy for DNA detection combining bio-barcode assay and metal-enhanced fluorescence modality.

Science.gov (United States)

Zhou, Zhenpeng; Li, Tian; Huang, Hongduan; Chen, Yang; Liu, Feng; Huang, Chengzhi; Li, Na

2014-11-11

Silver-enhanced fluorescence was coupled with a bio-barcode assay to facilitate a dual amplification assay to demonstrate a non-enzymatic approach for simple and sensitive detection of DNA. In the assay design, magnetic nanoparticles seeded with silver nanoparticles were modified with the capture DNA, and silver nanoparticles were modified with the binding of ssDNA and the fluorescently labeled barcode dsDNA. Upon introduction of the target DNA, a sandwich structure was formed because of the hybridization reaction. By simple magnetic separation, silver-enhanced fluorescence of barcode DNAs could be readily measured without the need of a further step to liberate barcode DNAs from silver nanoparticles, endowing the method with simplicity and high sensitivity with a detection limit of 1 pM.

Adaptation of the Nelson-Somogyi reducing-sugar assay to a microassay using microtiter plates.

Science.gov (United States)

Green, F; Clausen, C A; Highley, T L

1989-11-01

The Nelson-Somogyi assay for reducing sugars was adapted to microtiter plates. The primary advantages of this modified assay are (i) smaller sample and reagent volumes, (ii) elimination of boiling and filtration steps, (iii) automated measurement with a dual-wavelength scanning TLC densitometer, (iv) increased range and reproducibility, and (v) automated colorimetric readings by reflectance rather than absorbance.

Evaluation of three gentamicin serum assay techniques

International Nuclear Information System (INIS)

Matzke, G.R.; Gwizdala, C.; Wery, J.; Ferry, D.; Starnes, R.

1982-01-01

This investigation was designed to compare the enzyme-modified immunoassay (Syva--EMIT) with a radioimmunoassay (New England Nuclear--RIA) and the radiometric assay (Johnston--BACTEC) to determine the optimal assay for use in our aminoglycoside dosing service. The serum concentration determinations obtained via the three assay methods were analyzed by linear regression analysis. Significant positive correlations were noted between the three assay techniques (p less than 0.005) during both sample collection phases. The coefficients of determination for EMIT vs BACTEC and RIA vs BACTEC were 0.73 and 0.83 during phase 1, respectively, and 0.65 and 0.68 during phase 2, respectively. The slope of the regression lines also varied markedly during the two phases; 0.49 and 0.42 for EMIT and for RIA vs BACTEC, respectively, during phase 1 compound with 1.12 and 0.77, respectively, during phase 2. The differences noted in these relationships during phase 1 and 2 may be related to the alteration of the pH of the control sera utilized in the BACTEC assay. In contrast, RIA vs EMIT regression analysis indicated that existence of a highly significant relationship (p less than 0.0005 and r2 . 0.90). The EMIT technique was the easiest and most accurate for determination of serum gentamicin concentrations, whereas the BACTEC method was judged unacceptable for clinical use

Quantitative Correlation of in Vivo Properties with in Vitro Assay Results: The in Vitro Binding of a Biotin–DNA Analogue Modifier with Streptavidin Predicts the in Vivo Avidin-Induced Clearability of the Analogue-Modified Antibody

Science.gov (United States)

Dou, Shuping; Virostko, John; Greiner, Dale L.; Powers, Alvin C.; Liu, Guozheng

2016-01-01

Quantitative prediction of in vivo behavior using an in vitro assay would dramatically accelerate pharmaceutical development. However, studies quantitatively correlating in vivo properties with in vitro assay results are rare because of the difficulty in quantitatively understanding the in vivo behavior of an agent. We now demonstrate such a correlation as a case study based on our quantitative understanding of the in vivo chemistry. In an ongoing pretargeting project, we designed a trifunctional antibody (Ab) that concomitantly carried a biotin and a DNA analogue (hereafter termed MORF). The biotin and the MORF were fused into one structure prior to conjugation to the Ab for the concomitant attachment. Because it was known that avidin-bound Ab molecules leave the circulation rapidly, this design would theoretically allow complete clearance by avidin. The clearability of the trifunctional Ab was determined by calculating the blood MORF concentration ratio of avidin-treated Ab to non-avidin-treated Ab using mice injected with these compounds. In theory, any compromised clearability should be due to the presence of impurities. In vitro, we measured the biotinylated percentage of the Ab-reacting (MORF-biotin)⊃-NH2 modifier, by addition of streptavidin to the radiolabeled (MORF-biotin)⊃-NH2 samples and subsequent high-performance liquid chromatography (HPLC) analysis. On the basis of our previous quantitative understanding, we predicted that the clearability of the Ab would be equal to the biotinylation percentage measured via HPLC. We validated this prediction within a 3% difference. In addition to the high avidin-induced clearability of the trifunctional Ab (up to ~95%) achieved by the design, we were able to predict the required quality of the (MORF-biotin)⊃-NH2 modifier for any given in vivo clearability. This approach may greatly reduce the steps and time currently required in pharmaceutical development in the process of synthesis, chemical analysis, in

Characterization of modified allergen extracts by in vitro beta-hexosaminidase release from rat basophils.

Science.gov (United States)

Gehlhar, Kirsten; Peters, Marcus; Brockmann, Kirsten; van Schijndel, Hans; Bufe, Albrecht

2005-04-01

To date, there is no well-established test available that can be used to measure functional properties of modified allergens (allergoids). Due to the cross-linking process, the IgE-binding capacity of the allergens, normally necessary for their characterization, is lost. The aim of this study was to test whether the rat basophilic leukaemia (RBL) cell assay (beta-hexosaminidase release by rat basophils upon allergen stimulation) can be adopted to characterize allergoids and to evaluate the assay for testing allergoids and native allergens as well. Mice were immunized with native and modified Phleumpratense extracts in the presence of alum. Their sera were used to sensitize RBL-2H3 cells and measure basophil stimulation induced by different allergen extracts in the presence or absence of various additives. Sera containing specific IgE against both extract formulations were obtained. Native as well as modified extracts induced dose-dependent beta-hexosaminidase release from RBL cells. Both extracts were used to evaluate the characteristics of the assay, which showed high precision. Storage conditions were chosen to enhance extract degradation, which could be read directly from the altered stimulatory capacity of the extracts. Additives turned out to have diverse effects on the assay, whereas phenol had no measurable effect, alum had an inhibitory effect and glycerol elevated basophil activation. For the first time, a reliable, precise in vitro assay is available that is able to directly measure the properties of modified allergen extracts after their production process. The test is well evaluated and its advantages and limitations are discussed in this report. Copyright (c) 2005 S. Karger AG, Basel

Genotoxicity of cadmium chloride in the marine gastropod Nerita chamaeleon using comet assay and alkaline unwinding assay

Digital Repository Service at National Institute of Oceanography (India)

Sarkar, A.; Bhagat, J.; Ingole, B.S.; Rao, P.V.S.S.D.P.; Markad, V.L.

cell lines. Toxicology 126:103–114. Mitchelmore CL, Birmelin C, Chipman JK, Livingstone DR. 1998. Evidence for cytochrome P-450 catalysis and free radical involvement in the production of DNA strand breaks by benzo[a]pyrene and nitroaromatics... Lymnaea luteola L. Aquat Toxicol 124:83-90. Anderson D, Yu TW, Phillips BJ, Schmezer P. 1994. The effect of various antioxidants and other modifying agents on oxygen-radical-generated DNA damage in human lymphocytes in the Comet assay. Mutat Res...

A simple coated-tube assay for alpha-foeto protein for clinical use

International Nuclear Information System (INIS)

Dakubu, S.; Ahene, I.S.; Foli, A.K.

1977-01-01

A standard method for coating plastic tubes with antiserum has been applied to coat tubes with rabbit antiserum to human alpha-foeto protein. The coated plastic tubes have been used to set up a radioimmunoassay system which is sensitive and convenient for use on the occasional clinical sample. For a successful coated-tube assay, it was found necessary to modify the final incubation mixture from what was suitable in a standard double antibody assay system. (orig.) [de

T-S Fuzzy Modelling and H∞ Attitude Control for Hypersonic Gliding Vehicles

Directory of Open Access Journals (Sweden)

Weidong Zhang

2017-01-01

Full Text Available This paper addresses the T-S fuzzy modelling and H∞ attitude control in three channels for hypersonic gliding vehicles (HGVs. First, the control-oriented affine nonlinear model has been established which is transformed from the reentry dynamics. Then, based on Taylor’s expansion approach and the fuzzy linearization approach, the homogeneous T-S local modelling technique for HGVs is proposed. Given the approximation accuracy and controller design complexity, appropriate fuzzy premise variables and operating points of interest are selected to construct the T-S homogeneous submodels. With so-called fuzzy blending, the original plant is transformed into the overall T-S fuzzy model with disturbance. By utilizing Lyapunov functional approach, a state feedback fuzzy controller has been designed based on relaxed linear matrix inequality (LMI conditions to stable the original plants with a prescribed H∞ performance of disturbance. Finally, numerical simulations are performed to demonstrate the effectiveness of the proposed H∞ T-S fuzzy controller for the original attitude dynamics; the superiority of the designed T-S fuzzy controller compared with other local controllers based on the constructed fuzzy model is shown as well.

Rapid colorimetric assay for gentamicin injection.

Science.gov (United States)

Tarbutton, P

1987-01-01

A rapid colorimetric method for determining gentamicin concentration in commercial preparations of gentamicin sulfate injection was developed. Methods currently available for measuring gentamicin concentration via its colored complex with cupric ions in alkaline solution were modified to reduce the time required for a single analysis. The alkaline copper tartrate (ACT) reagent solution was prepared such that each milliliter contained 100 mumol cupric sulfate, 210 mumol potassium sodium tartrate, and 1.25 mmol sodium hydroxide. The assay involves mixing 0.3 mL gentamicin sulfate injection 40 mg/mL (of gentamicin), 1.0 mL ACT reagent, and 0.7 mL water; the absorbance of the resulting solution at 560 nm was used to calculate the gentamicin concentration in the sample. For injections containing 10 mg/mL of gentamicin, the amount of the injection was increased to 0.5 mL and water decreased to 0.5 mL. The concentration of gentamicin in samples representing 11 lots of gentamicin sulfate injection 40 mg/mL and 8 lots of gentamicin sulfate injection 10 mg/mL was determined. The specificity, reproducibility, and accuracy of the assay were assessed. The colored complex was stable for at least two hours. Gentamicin concentration ranged from 93.7 to 108% and from 95 to 109% of the stated label value of the 40 mg/mL and the 10 mg/mL injections, respectively. No components of the preservative system present in the injections interfered with the assay. Since other aminoglycosides produced a colored complex, the assay is not specific for gentamicin. The assay was accurate and reproducible over the range of 4-20 mg of gentamicin. This rapid and accurate assay can be easily applied in the hospital pharmacy setting.

Tracheal intubation in the emergency department: a comparison of GlideScope® video laryngoscopy to direct laryngoscopy in 822 intubations.

Science.gov (United States)

Sakles, John C; Mosier, Jarrod M; Chiu, Stephen; Keim, Samuel M

2012-04-01

Video laryngoscopy has, in recent years, become more available to emergency physicians. However, little research has been conducted to compare their success to conventional direct laryngoscopy. To compare the success rates of GlideScope(®) (Verathon Inc., Bothell, WA) videolaryngoscopy (GVL) with direct laryngoscopy (DL) for emergency department (ED) intubations. This was a 24-month retrospective observational study of all patients intubated in a single academic ED with a level I trauma center. Structured data forms were completed after each intubation and entered into a continuous quality improvement database. All patients intubated in the ED with either the GlideScope(®) standard, Cobalt, Ranger, or traditional Macintosh or Miller laryngoscopes were included. All patients intubated before arrival were excluded. Primary analysis evaluated overall and first-attempt success rates, operator experience level, performance characteristics of GVL, complications, and reasons for failure. There were 943 patients intubated during the study period; 120 were excluded due to alternative management strategies. DL was used in 583 (62%) patients, and GVL in 360 (38%). GVL had higher first-attempt success (75%, p = 0.03); DL had a higher success rate when more than one attempt was required (57%, p = 0.003). The devices had statistically equivalent overall success rates. GVL had fewer esophageal intubations (n = 1) than DL (n = 18); p = 0.005. The two techniques performed equivalently overall, however, GVL had a higher overall success rate, and lower number of esophageal complications. In the setting of ED intubations, GVL offers an excellent option to maximize first-attempt success for airway management. Published by Elsevier Inc.

Radioligand binding assay of cortisol using horse transcortin

International Nuclear Information System (INIS)

Dash, R.J.; Sharma, B.R.; Lata, V.

1979-01-01

A modified radioligand binding assay was developed to measure cortisol in a single methylene chloride extract of human plasma/serum. The assay utilises 5 percent horse serum for cortisol binding and 3 H-cortisol as tracer. Except for a cross reaction of 13.3 percent for cortisone and 7 percent for prednisolone that for other steroids tested was negligible. The assay sensitivity at the lower 95 percent inhibition of buffer controls was 20 pg/tube. The log dose logit response standard curve was linear between 80 pg and 5 ng/tube. Recovery(Y) of cortisol added (x) to male and pregnant female plasma was quantitated (y = 0.983 X-0.47, r = 0.98). Regression analysis of cortisol estimates obtained in 51 plasma/serum samples with this assay system and a specific radioimmunoassay (using cortisol-3-BSA antiserum) for plasma cortisol gave a coefficient of correlation (r) of 0.95 and a regression coefficient (b) of 0.97. The method was found to be simple and highly reproducible. Availability of all reagents in the aqueous phase permitted handling of a large number of samples by a single technician. (auth.)

DNA-repair measurements by use of the modified comet assay

DEFF Research Database (Denmark)

Godschalk, Roger W L; Ersson, Clara; Riso, Patrizia

2013-01-01

The measurement of DNA-repair activity by extracts from cells or tissues by means of the single-cell gel electrophoresis (comet) assay has a high potential to become widely used in biomonitoring studies. We assessed the inter-laboratory variation in reported values of DNA-repair activity...... on substrate cells that had been incubated with Ro19-8022 plus light to generate oxidatively damaged DNA. Eight laboratories assessed the DNA-repair activity of three cell lines (i.e. one epithelial and two fibroblast cell lines), starting with cell pellets or with cell extracts provided by the coordinating...... laboratory. There was a large inter-laboratory variation, as evidenced by the range in the mean level of repair incisions between the laboratory with the lowest (0.002incisions/10(6)bp) and highest (0.988incisions/10(6)bp) incision activity. Nevertheless, six out of eight laboratories reported the same cell...

Detection of bovine herpesvirus 4 glycoprotein B and thymidine kinase DNA by PCR assays in bovine milk

NARCIS (Netherlands)

Wellenberg, G.J.; Verstraten, E.; Belak, S.; Verschuren, S.B.E.; Rijsewijk, F.A.M.; Peshev, R.; Oirschot, van J.T.

2001-01-01

A polymerase chain reaction (PCR) assay was developed to detect bovine herpesvirus 4 (BHV4) glycoprotein B (gB) DNA, and a nested-PCR assay was modified for the detection of BHV4 thymidine kinase (TK) DNA in bovine milk samples. To identify false-negative PCR results, internal control templates were

Interlaboratory validation of the modified murine local lymph node assay based on adenosine triphosphate measurement.

Science.gov (United States)

Omori, Takashi; Idehara, Kenji; Kojima, Hajime; Sozu, Takashi; Arima, Kazunori; Goto, Hirohiko; Hanada, Tomohiko; Ikarashi, Yoshiaki; Inoda, Taketo; Kanazawa, Yukiko; Kosaka, Tadashi; Maki, Eiji; Morimoto, Takashi; Shinoda, Shinsuke; Shinoda, Naoki; Takeyoshi, Masahiro; Tanaka, Masashi; Uratani, Mamoru; Usami, Masahito; Yamanaka, Atsushi; Yoneda, Tomofumi; Yoshimura, Isao; Yuasa, Atsuko

2008-01-01

The murine local lymph node assay (LLNA) is a well-established alternative to the guinea pig maximization test (GPMT) or Buehler test (BT) for the assessment of the skin sensitizing ability of drugs and chemicals. Daicel Chemical Industries Ltd. has developed a modified LLNA based on the adenosine triphosphate (ATP) content (LLNA-DA). We conducted 2 interlaboratory validation studies to evaluate the reliability and relevance of LLNA-DA. The experiment involved 17 laboratories, wherein 14 chemicals were examined under blinded conditions. In the first study, 3 chemicals were examined in 10 laboratories and the remaining 9 were examined in 3 laboratories. In the second study, 1 chemical was examined in 7 laboratories and the remaining 4 chemicals were examined in 4 laboratories. The data were expressed as the ATP content for each chemical-treated group, and the stimulation index (SI) for each chemical-treated group was determined as the increase in the ATP content relative to the concurrent vehicle control group. An SI of 3 was set as the cut-off value for exhibiting skin sensitization activity. The results of the first study obtained in the experiments conducted for the 3 chemicals that were examined in all the 10 laboratories and for 5 of the remaining 9 chemicals were sufficiently consistent with small variations in their SI values. The sensitivity, specificity, and accuracy of LLNA-DA against those of GPMT/BT were 7/8 (87.5%), 3/3 (100%), and 10/11 (90.9%), respectively. In the second study, all the 5 chemicals studied demonstrated acceptably small interlaboratory variations. In the first study, a large variation was observed for 2 chemicals; in the second study, this variation was small. It was attributed to the application of dimethylsulfoxide as the solvent for the metallic salts. In conclusion, these 2 studies provide good evidence for the reliability of the LLNA-DA.

A facile molecularly imprinted polymer-based fluorometric assay for detection of histamine

DEFF Research Database (Denmark)

Feng, Xiaotong; Ashley, Jon; Zhou, Tongchang

2018-01-01

urgently needed. In this paper, we developed a facile and cost-effective molecularly imprinted polymer (MIP)-based fluorometric assay to directly quantify histamine. Histamine-specific MIP nanoparticles (nanoMIPs) were synthesized using a modified solid-phase synthesis method. They were then immobilized...

Analysis of chromium-51 release assay data using personal computer spreadsheet software

International Nuclear Information System (INIS)

Lefor, A.T.; Steinberg, S.M.; Wiebke, E.A.

1988-01-01

The Chromium-51 release assay is a widely used technique to assess the lysis of labeled target cells in vitro. We have developed a simple technique to analyze data from Chromium-51 release assays using the widely available LOTUS 1-2-3 spreadsheet software. This package calculates percentage specific cytotoxicity and lytic units by linear regression. It uses all data points to compute the linear regression and can determine if there is a statistically significant difference between two lysis curves. The system is simple to use and easily modified, since its implementation requires neither knowledge of computer programming nor custom designed software. This package can help save considerable time when analyzing data from Chromium-51 release assays

Small unilamellar vesicles as reagents: a chemically defined, quantitative assay for lectins

Energy Technology Data Exchange (ETDEWEB)

Rando, R.R.

1981-01-01

Samll unilamellar vesicles containing synthetic glycolipids can be prepared. These vesicles are aggregated by the appropriate lectin (Orr et al., 1979; Rando and Bangerter, 1979; Slama and Rando, 1980). It is shown here that extent of aggregation of these vesicles as measured by light scattering at 360 nm, is, under certain conditions, linear with amount of lectin added. This forms the basis of a rapid and simple quantitative assay for lectins using the modified vesicles as a defined chemical substrate. The assay is sensitive to lectin concentrations in the low ..mu..g range. The assay is applied here to studies on concanavalin A, Ricinus communis agglutinin and the ..cap alpha..-fucosyl binding lectin from Ulex europaeus (Type I).

Investigation of the antibiofilm capacity of peptide-modified stainless steel.

Science.gov (United States)

Cao, Pan; Li, Wen-Wu; Morris, Andrew R; Horrocks, Paul D; Yuan, Cheng-Qing; Yang, Ying

2018-03-01

Biofilm formation on surfaces is an important research topic in ship tribology and medical implants. In this study, dopamine and two types of synthetic peptides were designed and attached to 304 stainless steel surfaces, aiming to inhibit the formation of biofilms. A combinatory surface modification procedure was applied in which dopamine was used as a coupling agent, allowing a strong binding ability with the two peptides. X-ray photoelectron spectroscopy (XPS), elemental analysis, contact angle measurement and surface roughness test were used to evaluate the efficiency of the peptide modification. An antibiofilm assay against Staphylococcus aureus was conducted to validate the antibiofilm capacity of the peptide-modified stainless steel samples. XPS analysis confirmed that the optimal dopamine concentration was 40 µg ml -1 in the coupling reaction. Element analysis showed that dopamine and the peptides had bound to the steel surfaces. The robustness assay of the modified surface demonstrated that most peptide molecules had bound on the surface of the stainless steel firmly. The contact angle of the modified surfaces was significantly changed. Modified steel samples exhibited improved antibiofilm properties in comparison to untreated and dopamine-only counterpart, with the peptide 1 modification displaying the best antibiofilm effect. The modified surfaces showed antibacterial capacity. The antibiofilm capacity of the modified surfaces was also surface topography sensitive. The steel sample surfaces polished with 600# sandpaper exhibited stronger antibiofilm capacity than those polished with other types of sandpapers after peptide modification. These findings present valuable information for future antifouling material research.

The effects of posterior talar glide with dorsiflexion of the ankle on mobility, muscle strength and balance in stroke patients: a randomised controlled trial

OpenAIRE

Lee, Jin; Kim, Ju-O; Lee, Byoung-Hee

2017-01-01

[Purpose] The purpose of this study was to examine the effectiveness of posterior talar glide (PTG) with dorsiflexion of the ankle on stroke patients ankle mobility, muscle strength, and balance ability. [Subjects and Methods] Thirty-four subjects were randomly assigned to either a PTG with dorsiflexion group (PTG; n=17), or a weight-bearing with placebo PTG group (control; n=17). Subjects in the PTG group performed PTG with dorsiflexion, designed to improve ankle mobility, muscle strength an...

Solid-phase peptide quantitation assay using labeled monoclonal antibody and glutaraldehyde fixation

International Nuclear Information System (INIS)

Kasprzyk, P.G.; Cuttitta, F.; Avis, I.; Nakanishi, Y.; Treston, A.; Wong, H.; Walsh, J.H.; Mulshine, J.L.

1988-01-01

A solid-phase radioimmunoassay utilizing iodinated peptide-specific monoclonal antibody as a detection system instead of labeled peptide has been developed. Regional specific monoclonal antibodies to either gastrin-releasing peptide or gastrin were used as models to validate the general application of our modified assay. Conditions for radioactive labeling of the monoclonal antibody were determined to minimize oxidant damage, which compromises the sensitivity of other reported peptide quantitation assays. Pretreatment of 96-well polyvinyl chloride test plates with a 5% glutaraldehyde solution resulted in consistent retention of sufficient target peptide on the solid-phase matrix to allow precise quantitation. This quantitative method is completed within 1 h of peptide solid phasing. Pretreatment of assay plates with glutaraldehyde increased binding of target peptide and maximized antibody binding by optimizing antigen presentation. The hypothesis that glutaraldehyde affects both peptide binding to the plate and orientation of the peptide was confirmed by analysis of several peptide analogs. These studies indicate that peptide binding was mediated through a free amino group leaving the carboxy-terminal portion of the target peptide accessible for antibody binding. It was observed that the length of the peptide also affects the amount of monoclonal antibody that will bind. Under the optimal conditions, results from quantitation of gastrin-releasing peptide in relevant samples agree well with those from previously reported techniques. Thus, we report here a modified microplate assay which may be generally applied for the rapid and sensitive quantitation of peptide hormones

Loop-mediated isothermal amplification (LAMP) method for detection of genetically modified maize T25.

Science.gov (United States)

Xu, Junyi; Zheng, Qiuyue; Yu, Ling; Liu, Ran; Zhao, Xin; Wang, Gang; Wang, Qinghua; Cao, Jijuan

2013-11-01

The loop-mediated isothermal amplification (LAMP) assay indicates a potential and valuable means for genetically modified organism (GMO) detection especially for its rapidity, simplicity, and low cost. We developed and evaluated the specificity and sensitivity of the LAMP method for rapid detection of the genetically modified (GM) maize T25. A set of six specific primers was successfully designed to recognize six distinct sequences on the target gene, including a pair of inner primers, a pair of outer primers, and a pair of loop primers. The optimum reaction temperature and time were verified to be 65°C and 45 min, respectively. The detection limit of this LAMP assay was 5 g kg(-1) GMO component. Comparative experiments showed that the LAMP assay was a simple, rapid, accurate, and specific method for detecting the GM maize T25.

Development of a loop-mediated isothermal amplification (LAMP) assay for rapid and specific detection of common genetically modified organisms (GMOs).

Science.gov (United States)

Feng, Jiawang; Tang, Shiming; Liu, Lideng; Kuang, Xiaoshan; Wang, Xiaoyu; Hu, Songnan; You, Shuzhu

2015-03-01

Here, we developed a loop-mediated isothermal amplification (LAMP) assay for 11 common transgenic target DNA in GMOs. Six sets of LAMP primer candidates for each target were designed and their specificity, sensitivity, and reproductivity were evaluated. With the optimized LAMP primers, this LAMP assay was simply run within 45-60 min to detect all these targets in GMOs tested. The sensitivity, specificity, and reproductivity of the LAMP assay were further analyzed in comparison with those of Real-Time PCR. In consistent with real-time PCR, detection of 0.5% GMOs in equivalent background DNA was possible using this LAMP assay for all targets. In comparison with real-time PCR, the LAMP assay showed the same results with simple instruments. Hence, the LAMP assay developed can provide a rapid and simple approach for routine screening as well as specific events detection of many GMOs.

Selective detection and quantification of modified DNA with solid-state nanopores.

Science.gov (United States)

Carlsen, Autumn T; Zahid, Osama K; Ruzicka, Jan A; Taylor, Ethan W; Hall, Adam R

2014-10-08

We demonstrate a solid-state nanopore assay for the unambiguous discrimination and quantification of modified DNA. Individual streptavidin proteins are employed as high-affinity tags for DNA containing a single biotin moiety. We establish that the rate of translocation events corresponds directly to relative concentration of protein-DNA complexes and use the selectivity of our approach to quantify modified oligonucleotides from among a background of unmodified DNA in solution.

One simple DNA extraction device and its combination with modified visual loop-mediated isothermal amplification for rapid on-field detection of genetically modified organisms.

Science.gov (United States)

Zhang, Miao; Liu, Yinan; Chen, Lili; Quan, Sheng; Jiang, Shimeng; Zhang, Dabing; Yang, Litao

2013-01-02

Quickness, simplicity, and effectiveness are the three major criteria for establishing a good molecular diagnosis method in many fields. Herein we report a novel detection system for genetically modified organisms (GMOs), which can be utilized to perform both on-field quick screening and routine laboratory diagnosis. In this system, a newly designed inexpensive DNA extraction device was used in combination with a modified visual loop-mediated isothermal amplification (vLAMP) assay. The main parts of the DNA extraction device included a silica gel membrane filtration column and a modified syringe. The DNA extraction device could be easily operated without using other laboratory instruments, making it applicable to an on-field GMO test. High-quality genomic DNA (gDNA) suitable for polymerase chain reaction (PCR) and isothermal amplification could be quickly isolated from plant tissues using this device within 15 min. In the modified vLAMP assay, a microcrystalline wax encapsulated detection bead containing SYBR green fluorescent dye was introduced to avoid dye inhibition and cross-contaminations from post-LAMP operation. The system was successfully applied and validated in screening and identification of GM rice, soybean, and maize samples collected from both field testing and the Grain Inspection, Packers, and Stockyards Administration (GIPSA) proficiency test program, which demonstrated that it was well-adapted to both on-field testing and/or routine laboratory analysis of GMOs.

Development of a qualitative real-time PCR method to detect 19 targets for identification of genetically modified organisms.

Science.gov (United States)

Peng, Cheng; Wang, Pengfei; Xu, Xiaoli; Wang, Xiaofu; Wei, Wei; Chen, Xiaoyun; Xu, Junfeng

2016-01-01

As the amount of commercially available genetically modified organisms (GMOs) grows recent years, the diversity of target sequences for molecular detection techniques are eagerly needed. Considered as the gold standard for GMO analysis, the real-time PCR technology was optimized to produce a high-throughput GMO screening method. With this method we can detect 19 transgenic targets. The specificity of the assays was demonstrated to be 100 % by the specific amplification of DNA derived from reference material from 20 genetically modified crops and 4 non modified crops. Furthermore, most assays showed a very sensitive detection, reaching the limit of ten copies. The 19 assays are the most frequently used genetic elements present in GM crops and theoretically enable the screening of the known GMO described in Chinese markets. Easy to use, fast and cost efficient, this method approach fits the purpose of GMO testing laboratories.

Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection

Science.gov (United States)

Paudel, Damodar; Jarman, Richard; Limkittikul, Kriengsak; Klungthong, Chonticha; Chamnanchanunt, Supat; Nisalak, Ananda; Gibbons, Robert; Chokejindachai, Watcharee

2011-01-01

Background: Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman® assay and conventional nested PCR assay. Aims: To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conventional nested PCR (modified Lanciotti). Materials and Methods: Eight cultured virus strains were diluted in tenth dilution down to undetectable level by the PCR to optimize the primer, temperature (annealing, and extension and to detect the limit of detection of the assay. Hundred and ninety three ELISA and PCR proved dengue clinical samples were tested with real time SYBR® Green assay, real time Taqman® assay to compare the sensitivity and specificity. Results: Sensitivity and specificity of real time SYBR® green dengue assay (84% and 66%, respectively) was almost comparable to those (81% and 74%) of Taqman real time PCR dengue assay. Real time SYBR® green RT-PCR was equally sensitive in primary and secondary infection while real time Taqman was less sensitive in the secondary infection. Sensitivity of real time Taqman on DENV3 (87%) was equal to SYBR green real time PCR dengue assay. Conclusion: We developed low cost rapid diagnostic SYBR green dengue assay. Further study is needed to make duplex primer assay for the serotyping of dengue virus. PMID:22363089

Ab initio modeling of plasticity in HCP metals: pure zirconium and titanium and effect of oxygen

International Nuclear Information System (INIS)

Chaari, Nermine

2015-01-01

We performed atomistic simulations to determine screw dislocations properties in pure zirconium and titanium and to explain the hardening effect attributed to oxygen alloying in both hexagonal close-packed transition metals. We used two energetic models: ab initio calculations based on the density functional theory and calculations with an empirical potential. The complete energetic profile of the screw dislocation when gliding in the different slip planes is obtained in pure Zr. Our calculations reveal the existence of a metastable configuration of the screw dislocation partially spread in the first order pyramidal plane. This configuration is responsible for the cross slip of screw dislocations from prismatic planes, the easiest glide planes, to pyramidal or basal planes. This energy profile is affected by oxygen addition. Ab initio calculations reveal two main effects: oxygen enhances pyramidal cross slip by modifying the dislocation core structure, and pins the dislocation in its metastable sessile configuration. The same modeling approach is applied to titanium. In pure Ti, the same configurations of the screw dislocation in Zr are obtained, but with different energy levels. This leads to a different gliding mechanism. The same way as in Zr, oxygen enhances pyramidal glide in Ti by modifying the dislocation core structure. Besides, oxygen atom lowers the energy of the metastable configuration but not enough to pin the dislocation in this sessile configuration. (author) [fr

Accuracy of molecular diagnostics in pemphigus and bullous pemphigoid: comparison of commercial and modified mosaic indirect immunofluorescence tests as well as enzyme-linked immunosorbent assays.

Science.gov (United States)

Gornowicz-Porowska, Justyna; Seraszek-Jaros, Agnieszka; Bowszyc-Dmochowska, Monika; Kaczmarek, Elżbieta; Pietkiewicz, Paweł; Bartkiewicz, Paweł; Dmochowski, Marian

2017-02-01

Pemphigus and bullous pemphigoid (BP) are identified by autoantibodies (abs) against desmoglein 1, 3 (DSG1/3) and BP180/BP230, respectively. A novel mosaic to indirect immunofluorescence (IIF) using purified BP180 recombinant proteins spotted on slide and transfected cells expressing BP230, DSG1, DSG3 is available. The commercial (IgG detection) and modified (IgG4 detection) mosaic for indirect immunofluorescence (IIFc - IIF commercial, IIFm - IIF modified) and IgG ELISAs were evaluated in pemphigus and bullous pemphigoid (BP) molecular diagnostics. To compare diagnostic accuracy of commercial (IgG detection) and modified (IgG4 detection) mosaic IIF assay and to examine the diagnostic value of ELISAs in relation to mosaic IIF in routine laboratory diagnostics of pemphigus and BP. Sera from 37 BP and 19 pemphigus patients were studied. Associations between tests were assessed using Fisher's exact test. There are associations between the positive/negative samples detected by IIFc with desmoglein1 (DSG1)/desmoglein3 (DSG3)/BP230 transfected cells and ELISAs and no association between anti-BP180 IgG detection by IIFc and ELISA. IIFm with DSG1 and DSG3 showed both 100% sensitivity and 100% and 78% specificity, respectively, and 100% and 83% positive predictive value in relation to IIFc. IIFm with BP230 had 87% specificity, 55% sensitivity, whereas IIFm with BP180 had a 100% sensitivity and 13% specificity in relation to IIFc. The IIFc with DSG1/DSG3/BP230 transfected cells, excluding BP180 spots, is an alternative method to ELISA in pemphigus/BP diagnostics. IgG4 antibodies, both pathogenically and diagnostically important, are inconsistently detectable with IIFm.

The influence of torque and manual glide path on the defect or separation rate of NiTi rotary instruments in root canal therapy

Directory of Open Access Journals (Sweden)

Zarrabi M

2010-01-01

Full Text Available Introduction: One of the effecting factors in prognosis of root canal therapy is accidental procedure as broken files that may be unpreventable. Many manufacturers have designed and marketed various electromotors that can control rotational speed and torque. On the other hand, some studies have recommended applying a manual glide path to diminish contact area between the file and canal walls. The purpose of this study was evaluation of the effect of torque and a manual glide path on defects as separation of Nickel-titanium (NiTi rotary files. Materials and Methods: This ex vivo randomized controlled trial study was carried out on 160 canals of human′s matured molars with mild curvature (15-338. After initial preparation of samples and checking for inclusion criteria, in first group, preparation was carried out with air-driven handpiece, and in group two, Endo IT was used as electromotor. In both groups, Mtwo files with simultaneous technique were used for preparation. Then all data were collected and analyzed with Mann Whitny, Mantel Cox, and t-test. Results: No significant differences between two groups (P < 0.05 were observed. Based on survival analysis, safety probability of files after preparation of nine canals is 64% in group one and 69.9% in group two. There was no significant differences between this safety probability in two groups (P = 0.272. Conclusion: Usage of torque control handpiece is not an important factor, comparing instrumentation technique.

Incidence of dentinal defects after preparation of severely curved root canals using the Reciproc single-file system with and without prior creation of a glide path.

Science.gov (United States)

Saber, S E D M; Schäfer, E

2016-11-01

To investigate the incidence of dentinal defects after preparation of severely curved root canals using the Reciproc single-file system with and without prior creation of a glide path. Mesial roots from extracted mandibular first molars were collected and scanned with CBCT to assess the morphology of the root canal systems. Three groups of 20 anatomically comparable specimens were generated. The control group was left unprepared, whilst the experimental groups were prepared with Reciproc R25 with and without a glide path (groups RG and R, respectively). Roots were then sectioned perpendicular to the long axis at 2, 4, 6, 8 and 10 mm from the apex, and coloured photographs of the sections at 40× were obtained. Two blinded examiners registered the presence of dentinal defects twice at 2-week interval. Data were statistically analysed using the Fisher exact and Cochran's Q tests. No defects were observed in the control group. The overall incidence of dentinal defects was 26% in group R and 24% in group RG, with no significant differences between them (P > 0.05). Dentinal defects occurred significantly more often in the middle and coronal thirds compared to the apical third of the canals (P files. © 2015 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Calibration and validation of the 14C-labelled polyethylene glycol-binding assay for tannins in tropical browse

International Nuclear Information System (INIS)

Mlambo, V.; Makkar, H.P.S.

2005-01-01

This study evaluates the radiolabelled polyethylene glycol (PEG)-binding procedure [Silanikove, N., Shinder, D., Gilboa, N., Eyal, M., Nitsan, Z., 1996. Polyethylene glycol-binding to plant samples as an assay for the biological effects of tannins: predicting the negative effects of tannins in Mediterranean browse on rumen degradation. J. Agric. Food Chem. 44, 3230-3234] for tannin analysis, using 27 tropical browse plants. In this method, the amount of PEG bound to a plant sample is assumed to be a reflection of its tannin content. The method was modified to exclude the use of non-tanniniferous substrate for estimating non-specific binding (NSB) in tannin-containing substrates. Non-specific binding values varied widely (0.4-2.8 mg PEG/100 mg DM tannin-free substrate) when the tannin-free substrate was changed from wheat straw to either rye grass or maize shoots. We therefore propose a modified radiolabelled PEG-binding method to estimate the level of PEG-binding (PEGb) to tannin-containing foliage without using tannin-free substrate to correct for non-specific binding. In this approach, incremental levels of each tanniniferous substrate were used to generate PEGb values. The resultant linear response was analysed and tannin activity was expressed as the slope of the response curve (PEGbSlope) observed for each substrate. The slope takes into account the non-specific binding in each substrate, thus PEGbSlope does not require correction for NSB using tannin-free samples. This approach improved the correlation between PEGb and the 125 I-labelled bovine serum albumin precipitation assay. Relationships between the modified PEG-binding assay and radiolabelled bovine serum albumin assay, in vitro tannin bioassay and colorimetric assays are presented. (author)

Platelet Lysate-Modified Porous Silicon Microparticles for Enhanced Cell Proliferation in Wound Healing Applications.

Science.gov (United States)

Fontana, Flavia; Mori, Michela; Riva, Federica; Mäkilä, Ermei; Liu, Dongfei; Salonen, Jarno; Nicoletti, Giovanni; Hirvonen, Jouni; Caramella, Carla; Santos, Hélder A

2016-01-13

The new frontier in the treatment of chronic nonhealing wounds is the use of micro- and nanoparticles to deliver drugs or growth factors into the wound. Here, we used platelet lysate (PL), a hemoderivative of platelets, consisting of a multifactorial cocktail of growth factors, to modify porous silicon (PSi) microparticles and assessed both in vitro and ex vivo the properties of the developed microsystem. PL-modified PSi was assessed for its potential to induce proliferation of fibroblasts. The wound closure-promoting properties of the microsystem were then assessed in an in vitro wound healing assay. Finally, the PL-modified PSi microparticles were evaluated in an ex vivo experiment over human skin. It was shown that PL-modified PSi microparticles were cytocompatible and enhanced the cell proliferation in different experimental settings. In addition, this microsystem promoted the closure of the gap between the fibroblast cells in the wound healing assay, in periods of time comparable with the positive control, and induced a proliferation and regeneration process onto the human skin in an ex vivo experiment. Overall, our results show that PL-modified PSi microparticles are suitable microsystems for further development toward applications in the treatment of chronic nonhealing wounds.

Morphology of single Shockley-type stacking faults generated by recombination enhanced dislocation glide in 4H-SiC

Science.gov (United States)

Matsuhata, Hirofumi; Sekiguchi, Takashi

2018-04-01

Morphology of single Shockley-type stacking faults (SFs) generated by recombination enhanced dislocation glide (REDG) in 4H-SiC are discussed and analysed. A complete set of the 12 different dissociated states of basal-plane dislocation loops is obtained using the crystallographic space group operations. From this set, six different double rhombic-shaped SFs are derived. These tables indicate the rules that connect shapes of SFs with the locations of partial dislocations having different core structures, the positions of slip planes in a unit cell, and the Burgers vectors of partial dislocations. We applied these tables for the analysis of SFs generated by the REDG effect reported in the past articles. Shapes, growing process of SFs and perfect dislocations for origins of SFs were well analysed systematically.

Detection of NT-pro BNP using fluorescent protein modified by streptavidin as a label in immunochromatographic assay

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Haixia Li

2016-12-01

Full Text Available A novel fluorescent immunochromatographic assay for the detection of NT-proBNP in human serum has been developed. Based on a sandwich-type immunoassay format, analytes in samples were captured by one monoclonal antibody labeled with fluorescent protein and “sandwiched” by another monoclonal antibody immobilized on the nitrocellulose membrane, the fluorescence and concentration of analytes were measured and then calculated by fluoroanalyzer. The fluorescent protein is a fusion protein and was prepared through the application of Streptavidin gene SA, β subunit cpcB of Phycocyanin, lyase alr0617, and phycoerythrobilin synthetase gene ho1, pebA, pebB for covalent binding. It is characterized with higher stability, good solubility in water and it is not easy to quench fluorescence. Take the advantages of fluorescent protein, the immunochromatographic assay exhibited a wide linear range for NT-proBNP from 200 pg ml−1 to 26,000 pg ml−1, with a detection limit of 47 pg ml−1 under optimal conditions. Compared with chemiluminescence immunoassay (CLIA, 131 human serum samples were analyzed and the correlation coefficient of the developed immunoassay was 0.978. These results demonstrated that fluorescent immunochromatographic assay is a more rapid, sensitive, specific method and could be developed into a platform for more biomarkers determination in clinical practice. Keywords: NT-pro BNP, Fluorescent protein, Immunochromatographic assay

Analysis of variola and vaccinia virus neutralization assays for smallpox vaccines.

Science.gov (United States)

Hughes, Christine M; Newman, Frances K; Davidson, Whitni B; Olson, Victoria A; Smith, Scott K; Holman, Robert C; Yan, Lihan; Frey, Sharon E; Belshe, Robert B; Karem, Kevin L; Damon, Inger K

2012-07-01

Possible smallpox reemergence drives research for third-generation vaccines that effectively neutralize variola virus. A comparison of neutralization assays using different substrates, variola and vaccinia (Dryvax and modified vaccinia Ankara [MVA]), showed significantly different 90% neutralization titers; Dryvax underestimated while MVA overestimated variola neutralization. Third-generation vaccines may rely upon neutralization as a correlate of protection.

Paper-Based Digital Microfluidic Chip for Multiple Electrochemical Assay Operated by a Wireless Portable Control System

DEFF Research Database (Denmark)

Ruecha, Nipapan; Lee, Jumi; Chae, Heedo

2017-01-01

for multiple analysis assays are fabricated by affordable printing techniques. For enhanced sensitivity of the sensor, the working electrode is modified through the electrochemical method, namely by reducing graphene with voltammetry and coating gold nanoparticles by amperometry. Detachable sensor and absorber...... designed portable power supply and wireless control system, the active paper-based chip platform can be utilized as an advanced point-of-care device for multiple assays in digital microfluidics....

Evaluation of the reliability of maize reference assays for GMO quantification.

Science.gov (United States)

Papazova, Nina; Zhang, David; Gruden, Kristina; Vojvoda, Jana; Yang, Litao; Buh Gasparic, Meti; Blejec, Andrej; Fouilloux, Stephane; De Loose, Marc; Taverniers, Isabel

2010-03-01

A reliable PCR reference assay for relative genetically modified organism (GMO) quantification must be specific for the target taxon and amplify uniformly along the commercialised varieties within the considered taxon. Different reference assays for maize (Zea mays L.) are used in official methods for GMO quantification. In this study, we evaluated the reliability of eight existing maize reference assays, four of which are used in combination with an event-specific polymerase chain reaction (PCR) assay validated and published by the Community Reference Laboratory (CRL). We analysed the nucleotide sequence variation in the target genomic regions in a broad range of transgenic and conventional varieties and lines: MON 810 varieties cultivated in Spain and conventional varieties from various geographical origins and breeding history. In addition, the reliability of the assays was evaluated based on their PCR amplification performance. A single base pair substitution, corresponding to a single nucleotide polymorphism (SNP) reported in an earlier study, was observed in the forward primer of one of the studied alcohol dehydrogenase 1 (Adh1) (70) assays in a large number of varieties. The SNP presence is consistent with a poor PCR performance observed for this assay along the tested varieties. The obtained data show that the Adh1 (70) assay used in the official CRL NK603 assay is unreliable. Based on our results from both the nucleotide stability study and the PCR performance test, we can conclude that the Adh1 (136) reference assay (T25 and Bt11 assays) as well as the tested high mobility group protein gene assay, which also form parts of CRL methods for quantification, are highly reliable. Despite the observed uniformity in the nucleotide sequence of the invertase gene assay, the PCR performance test reveals that this target sequence might occur in more than one copy. Finally, although currently not forming a part of official quantification methods, zein and SSIIb

Calibration and validation of the {sup 14}C-labelled polyethylene glycol-binding assay for tannins in tropical browse

Energy Technology Data Exchange (ETDEWEB)

Mlambo, V. [Animal Production Unit, FAO/IAEA Agriculture and Biotechnology Laboratory, Seibersdorf (Austria)]. E-mail: vmlambo@agric.uniswa.sz; Makkar, H.P.S. [Animal Production and Health Section, Joint FAO/IAEA Division of Nuclear Techniques in Agriculture and Food, International Atomic Energy Agency, Vienna (Austria)

2005-08-19

This study evaluates the radiolabelled polyethylene glycol (PEG)-binding procedure [Silanikove, N., Shinder, D., Gilboa, N., Eyal, M., Nitsan, Z., 1996. Polyethylene glycol-binding to plant samples as an assay for the biological effects of tannins: predicting the negative effects of tannins in Mediterranean browse on rumen degradation. J. Agric. Food Chem. 44, 3230-3234] for tannin analysis, using 27 tropical browse plants. In this method, the amount of PEG bound to a plant sample is assumed to be a reflection of its tannin content. The method was modified to exclude the use of non-tanniniferous substrate for estimating non-specific binding (NSB) in tannin-containing substrates. Non-specific binding values varied widely (0.4-2.8 mg PEG/100 mg DM tannin-free substrate) when the tannin-free substrate was changed from wheat straw to either rye grass or maize shoots. We therefore propose a modified radiolabelled PEG-binding method to estimate the level of PEG-binding (PEGb) to tannin-containing foliage without using tannin-free substrate to correct for non-specific binding. In this approach, incremental levels of each tanniniferous substrate were used to generate PEGb values. The resultant linear response was analysed and tannin activity was expressed as the slope of the response curve (PEGbSlope) observed for each substrate. The slope takes into account the non-specific binding in each substrate, thus PEGbSlope does not require correction for NSB using tannin-free samples. This approach improved the correlation between PEGb and the {sup 125}I-labelled bovine serum albumin precipitation assay. Relationships between the modified PEG-binding assay and radiolabelled bovine serum albumin assay, in vitro tannin bioassay and colorimetric assays are presented. (author)

Quantum Dot-Based Luminescent Oxygen Channeling Assay for Potential Application in Homogeneous Bioassays.

Science.gov (United States)

Zhuang, Si-Hui; Guo, Xin-Xin; Wu, Ying-Song; Chen, Zhen-Hua; Chen, Yao; Ren, Zhi-Qi; Liu, Tian-Cai

2016-01-01

The unique photoproperties of quantum dots are promising for potential application in bioassays. In the present study, quantum dots were applied to a luminescent oxygen channeling assay. The reaction system developed in this study was based on interaction of biotin with streptavidin. Carboxyl-modified polystyrene microspheres doped with quantum dots were biotinylated and used as acceptors. Photosensitizer-doped carboxyl-modified polystyrene microspheres were conjugated with streptavidin and used as donors. The results indicated that the singlet oxygen that was released from the donor beads diffused into the acceptor beads. The acceptor beads were then exited via thioxene, and were subsequently fluoresced. To avoid generating false positives, a high concentration (0.01 mg/mL) of quantum dots is required for application in homogeneous immunoassays. Compared to a conventional luminescent oxygen channeling assay, this quantum dots-based technique requires less time, and would be easier to automate and miniaturize because it requires no washing to remove excess labels.

Factors affecting a cyanogen bromide-based assay of thiamin.

Science.gov (United States)

Wyatt, D T; Lee, M; Hillman, R E

1989-11-01

We analyzed extensively a modified thiochrome method for thiamin analysis. Acid phosphatase (EC 3.1.3.2) from potato was superior to either alpha-amylase or acid phosphatase from wheat germ as a dephosphorylating agent. Timing of cyanogen bromide exposure was important, but the assay had good precision and accuracy. The standard curve was linear from 10 to 3000 nmol/L. The within-run and between-run coefficients of variation for total thiamin in whole blood were 3.6% and 7.4%, respectively. Analytical recoveries for low, intermediate, and high additions of thiamin to whole blood were 93-109%. Sample yield was increased by 41% (+/- 29% SD) with pre-assay freezing. Samples were stable for two days at room temperature, for seven days when refrigerated, and for two years when frozen. Previously unreported interference was seen with penicillin derivatives, and with several commonly used diuretic and antiepileptic medications. This assay may be suitable for population screening; 200 samples could be analyzed weekly at a cost of +0.20 per sample.

Accuracy of molecular diagnostics in pemphigus and bullous pemphigoid: comparison of commercial and modified mosaic indirect immunofluorescence tests as well as enzyme-linked immunosorbent assays

Directory of Open Access Journals (Sweden)

Justyna Gornowicz-Porowska

2017-02-01

Full Text Available Introduction : Pemphigus and bullous pemphigoid (BP are identified by autoantibodies (abs against desmoglein 1, 3 (DSG1/3 and BP180/BP230, respectively. A novel mosaic to indirect immunofluorescence (IIF using purified BP180 recombinant proteins spotted on slide and transfected cells expressing BP230, DSG1, DSG3 is available. The commercial (IgG detection and modified (IgG4 detection mosaic for indirect immunofluorescence (IIFc – IIF commercial, IIFm – IIF modified and IgG ELISAs were evaluated in pemphigus and bullous pemphigoid (BP molecular diagnostics. Aim : To compare diagnostic accuracy of commercial (IgG detection and modified (IgG4 detection mosaic IIF assay and to examine the diagnostic value of ELISAs in relation to mosaic IIF in routine laboratory diagnostics of pemphigus and BP. Material and methods : Sera from 37 BP and 19 pemphigus patients were studied. Associations between tests were assessed using Fisher’s exact test. Results: There are associations between the positive/negative samples detected by IIFc with desmoglein1 (DSG1/desmoglein3 (DSG3/BP230 transfected cells and ELISAs and no association between anti-BP180 IgG detection by IIFc and ELISA. IIFm with DSG1 and DSG3 showed both 100% sensitivity and 100% and 78% specificity, respectively, and 100% and 83% positive predictive value in relation to IIFc. IIFm with BP230 had 87% specificity, 55% sensitivity, whereas IIFm with BP180 had a 100% sensitivity and 13% specificity in relation to IIFc. Conclusions : The IIFc with DSG1/DSG3/BP230 transfected cells, excluding BP180 spots, is an alternative method to ELISA in pemphigus/BP diagnostics. IgG4 antibodies, both pathogenically and diagnostically important, are inconsistently detectable with IIFm.

Study of the effect of thiols on the vasodilatory potency of S-nitrosothiols by using a modified aortic ring assay

International Nuclear Information System (INIS)

Giustarini, Daniela; Tsikas, Dimitrios; Rossi, Ranieri

2011-01-01

Both low-molecular-mass thiols (LMM-SH) and protein thiols (P-SH) can modulate the biological activity of S-nitrosothiols (RSNO) via S-transnitrosation reactions. It has been difficult to evaluate the entity of this effect in blood circulation by in vitro assays with isolated aorta rings so far, because media rich in proteins cannot be used due to the foaming as a consequence of the needed gas bubbling. We have modified the original apparatus for organ bioassay in order to minimize foaming and to increase analytical performance. By using this modified bioassay we investigated the vasodilatory potency of various endogenous RSNOs in the presence of physiologically relevant concentrations of albumin and LMM-SH. Our results show that the sulfhydryl group of the cysteine moiety of albumin and LMM-SH has a dramatic effect on the vasodilatory potency of RSNO. Considering the equilibrium constants for S-transnitrosation reactions and the concentration of P-SH and LMM-SH we measured in healthy humans (aged 18-85 years), we infer that the age-dependency of hematic levels of LMM-SH may have a considerable impact in RSNO-mediated vasodilation. S-Nitrosoproteins such as S-nitrosoalbumin may constitute a relatively silent and constant amount of circulating RSNO. On the other hand, LMM-SH may mediate and control the biological actions of S-nitrosoproteins via S-transnitrosation reactions, by forming more potent nitric oxide-releasing LMM-S-nitrosothiols. Lifestyle habits, status of health and individual age are proven factors that, in turn, may influence the concentration of these compounds. These aspects should be taken into consideration when testing the vasodilatory effects of RSNO in pre-clinical studies. - Highlights: → A modification of the organ chamber apparatus for aortic ring bioassays is proposed. → The new apparatus can work in the presence of albumin at physiological concentrations. → Potency of RSNOs was studied in the presence of albumin and low molecular

Computed micro-tomographic evaluation of glide path with nickel-titanium rotary PathFile in maxillary first molars curved canals.

Science.gov (United States)

Pasqualini, Damiano; Bianchi, Caterina Chiara; Paolino, Davide Salvatore; Mancini, Lucia; Cemenasco, Andrea; Cantatore, Giuseppe; Castellucci, Arnaldo; Berutti, Elio

2012-03-01

X-ray computed micro-tomography scanning allows high-resolution 3-dimensional imaging of small objects. In this study, micro-CT scanning was used to compare the ability of manual and mechanical glide path to maintain the original root canal anatomy. Eight extracted upper first permanent molars were scanned at the TOMOLAB station at ELETTRA Synchrotron Light Laboratory in Trieste, Italy, with a microfocus cone-beam geometry system. A total of 2,400 projections on 360° have been acquired at 100 kV and 80 μA, with a focal spot size of 8 μm. Buccal root canals of each specimen (n = 16) were randomly assigned to PathFile (P) or stainless-steel K-file (K) to perform glide path at the full working length. Specimens were then microscanned at the apical level (A) and at the point of the maximum curvature level (C) for post-treatment analyses. Curvatures of root canals were classified as moderate (≤35°) or severe (≥40°). The ratio of diameter ratios (RDRs) and the ratio of cross-sectional areas (RAs) were assessed. For each level of analysis (A and C), 2 balanced 2-way factorial analyses of variance (P < .05) were performed to evaluate the significance of the instrument factor and of canal curvature factor as well as the interactions of the factors both with RDRs and RAs. Specimens in the K group had a mean curvature of 35.4° ± 11.5°; those in the P group had a curvature of 38° ± 9.9°. The instrument factor (P and K) was extremely significant (P < .001) for both the RDR and RA parameters, regardless of the point of analysis. Micro-CT scanning confirmed that NiTi rotary PathFile instruments preserve the original canal anatomy and cause less canal aberrations. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

A histidine-rich protein 2-based malaria drug sensitivity assay for field use.

Science.gov (United States)

Noedl, Harald; Attlmayr, Bernhard; Wernsdorfer, Walther H; Kollaritsch, Herwig; Miller, Robert S

2004-12-01

With the spread of antimalarial drug resistance, simple and reliable tools for the assessment of antimalarial drug resistance, particularly in endemic regions and under field conditions, have become more important than ever before. We therefore developed a histidine-rich protein 2 (HRP2)-based drug sensitivity assay for testing of fresh isolates of Plasmodium falciparum in the field. In contrast to the HRP2 laboratory assay, the field assay uses a procedure that further simplifies the handling and culturing of malaria parasites by omitting centrifugation, washing, the use of serum, and dilution with uninfected red blood cells. A total of 40 fresh Plasmodium falciparum isolates were successfully tested for their susceptibility to dihydroartemisinin, mefloquine, quinine, and chloroquine (50% inhibitory concentration [IC50] = 3.43, 61.89, 326.75, and 185.31 nM, respectively). Results very closely matched those obtained with a modified World Health Organization schizont maturation assay (R2 = 0.96, P < 0.001; mean log difference at IC50 = 0.054).

Calibration of a Modified Californium Shuffler

International Nuclear Information System (INIS)

Sadowski, E.T.; Armstrong, F.; Oldham, R.; Ceo, R.; Williams, N.

1995-01-01

A californium shuffler originally designed to assay hollow cylindrical pieces of UA1 has been modified to assay solid cylinders. Calibration standards were characterized via chemical analysis of the molten UA1 taken during casting of the standards. The melt samples yielded much more reliable characterization data than drill samples taken from standards after the standards had solidified. By normalizing one well-characterized calibration curve to several standards at different enrichments, a relatively small number of standards was required to develop an enrichment-dependent calibration. The precision of this shuffler is 0.65%, and the typical random and systematic uncertainties are 0.53% and 0.73%, respectively, for a six minute assay of an ingot containing approximately 700 grams of 235 U. This paper will discuss (1) the discrepancies encountered when UA1 standards were characterized via melt samples versus drill samples, (2) a calibration methodology employing a small number of standards, and (3) a comparison of results from a previously unused shuffler with an existing shuffler. A small number of UA1 standards have been characterized using samples from the homogeneous molten state and have yielded enrichment-dependent and enrichment-independent calibration curves on two different shufflers

ABTS assay of phenol oxidase activity in soil.

Science.gov (United States)

Floch, Carine; Alarcon-Gutiérrez, Enrique; Criquet, Stéven

2007-12-01

Phenol oxidases (PO) are involved in degradation of many recalcitrant aromatic compounds and may be sensitive to some pollutants. Hence, their activities may be a useful indicator for evaluating soil quality and health. To this end, the aim of this study was to develop a simple method to assay PO activity directly in bulk samples by spectrophotometric test using 2,2'-azinobis-(-3 ethylbenzothiazoline-6-sulfononic acid) diammonium salt (ABTS) as the substrate. Three Mediterranean soils were used as models. For each soil, we studied the kinetic parameters and the effects of certain factors (i.e. amount of soil, pH, temperature, incubation time and substrate concentration) in order to determine the optimum conditions for the ABTS assay. Results showed that PO attain their optimum activities when incubating 0.1 g of soil at 30 degrees C for 5 min with 10 ml of a Modified Universal Buffer (MUB) at pH 2 and 200 microl of a 0.1 M ABTS solution.

Neutralization of X4- and R5-tropic HIV-1 NL4-3 variants by HOCl-modified serum albumins

Directory of Open Access Journals (Sweden)

Schwalbe Birco

2010-06-01

Full Text Available Abstract Background Myeloperoxidase (MPO, an important element of the microbicidal activity of neutrophils, generates hypochlorous acid (HOCl from H2O2 and chloride, which is released into body fluids. Besides its direct microbicidal activity, HOCl can react with amino acid residues and HOCl-modified proteins can be detected in vivo. Findings This report is based on binding studies of HOCl-modified serum albumins to HIV-1 gp120 and three different neutralization assays using infectious virus. The binding studies were carried out by surface plasmon resonance spectroscopy and by standard ELISA techniques. Virus neutralization assays were carried out using HIV-1 NL4-3 virus and recombinant strains with CXCR4 and CCR5 coreceptor usage. Viral infection was monitored by a standard p24 or X-gal staining assay. Our data demonstrate that HOCl-modified mouse-, bovine- and human serum albumins all bind to the HIV-1 NL4-3 gp120 (LAV glycoprotein in contrast to non-modified albumin. Binding of HOCl-modified albumin to gp120 correlated to the blockade of CD4 as well as that of V3 loop specific monoclonal antibody binding. In neutralization experiments, HOCl-modified serum albumins inhibited replication and syncytium formation of the X4- and R5-tropic NL4-3 isolates in a dose dependent manner. Conclusions Our data indicate that HOCl-modified serum albumin veils the binding site for CD4 and the V3 loop on gp120. Such masking of the viral gp120/gp41 envelope complex might be a simple but promising strategy to inactivate HIV-1 and therefore prevent infection when HOCl-modified serum albumin is applied, for example, as a topical microbicide.

Cartilage formation measured by a novel PIINP assay suggests that IGF-I does not stimulate but maintains cartilage formation ex vivo

DEFF Research Database (Denmark)

Madsen, S H; Sondergaard, B C; Jensen, Anne-Christine Bay

2009-01-01

explants were cultured in Dulbecco's modified Eagle's medium (DMEM):F12 in the presence of 0, 0.01, 0.1, 1, 10, or 100 ng/mL of IGF-I. The viability of the chondrocytes was measured by the colorimetric Alamar blue assay. Collagen formation was assessed from the conditioned medium by the PIINP assay...

Comparison of the reusable standard GlideScope® video laryngoscope and the disposable cobalt GlideScope® video laryngoscope for tracheal intubation in an academic emergency department: a retrospective review.

Science.gov (United States)

Sakles, John C; Patanwala, Asad E; Mosier, Jarrod; Dicken, John; Holman, Nathan

2014-04-01

The objective was to compare the first-pass success and clinical performance characteristics of the reusable standard GlideScope® video laryngoscope (sGVL) and the disposable Cobalt GlideScope® video laryngoscope (cGVL). This was a retrospective analysis of prospectively collected data recorded into a continuous quality improvement database at an urban academic emergency department (ED). The intent of the database is to evaluate operator performance and to track practice patterns used for intubation in the ED. Between July 1, 2007, and June 30, 2013, operators recorded all consecutive intubations performed in the ED. The database included patient demographics and detailed information about each intubation, such as device(s) used, reason for device selection, method of intubation, difficult airway characteristics, number of intubation attempts, and outcome of each attempt. The operator also evaluated the presence of lens fogging and extent of lens contamination. The primary outcome measure was first-pass success. Secondary outcome measures were ultimate success, Cormack-Lehane (CL) view of the airway, presence of lens fogging, and extent of lens contamination. Only adult patients age 18 years or older intubated with the sGVL or cGVL using a stylet, and who had data forms completed at the time of intubation, were included in this study. A total of 583 intubations were included in the study, 504 with the sGVL and 79 with cGVL. First pass success was achieved in 81.0% (95% confidence interval [CI]=77.3% to 84.3%) of patients in the sGVL group and in 58.2% (95% CI=46.6% to 69.2%) of patients in the cGVL group. In a multivariate logistic regression analysis, the sGVL was associated with a higher first pass success than the cGVL (odds ratio [OR]=3.3, 95% CI=1.9 to 5.8). The ultimate success of the sGVL was 92.1% (95% CI=89.4% to 94.3%) and the cGVL was 72.2% (95% CI=60.9% to 81.7%). A CL grade I or II view was obtained in 93.2% (95% CI=90.7% to 95.3%) in the sGVL group

Validation of a newly developed hexaplex real-time PCR assay for screening for presence of GMOs in food, feed and seed.

Science.gov (United States)

Bahrdt, C; Krech, A B; Wurz, A; Wulff, D

2010-03-01

For years, an increasing number and diversity of genetically modified plants has been grown on a commercial scale. The need for detection and identification of these genetically modified organisms (GMOs) calls for broad and at the same time flexible high throughput testing methods. Here we describe the development and validation of a hexaplex real-time polymerase chain reaction (PCR) screening assay covering more than 100 approved GMOs containing at least one of the GMO targets of the assay. The assay comprises detection systems for Cauliflower Mosaic Virus 35S promoter, Agrobacterium tumefaciens NOS terminator, Figwort Mosaic Virus 34S promoter and two construct-specific sequences present in novel genetically modified soybean and maize that lack common screening elements. Additionally a detection system for an internal positive control (IPC) indicating the presence or absence of PCR inhibiting substances was included. The six real-time PCR systems were allocated to five detection channels showing no significant crosstalk between the detection channels. As part of an extensive validation, a limit of detection (LOD(abs)) GMOs in processed and unprocessed food, feed and seed samples with high efficiency.

Association of a new type of gliding, filamentous, purple phototrophic bacterium inside bundles of Microcoleus chthonoplastes in hypersaline cyanobacterial mats

Science.gov (United States)

D'Amelio, E. D.; Cohen, Y.; Des Marais, D. J.

1987-01-01

An unidentified filamentous purple bacterium, probably belonging to a new genus or even a new family, is found in close association with the filamentous, mat-forming cyanobacterium Microcoleus chthonoplastes in a hypersaline pond at Guerrero Negro, Baja California Sur, Mexico, and in Solar Lake, Sinai, Egypt. This organism is a gliding, segmented trichome, 0.8-0.9 micrometer wide. It contains intracytoplasmic stacked lamellae which are perpendicular and obliquely oriented to the cell wall, similar to those described for the purple sulfur bacteria Ectothiorhodospira. These bacteria are found inside the cyanobacterial bundle, enclosed by the cyanobacterial sheath. Detailed transmission electron microscopical analyses carried out in horizontal sections of the upper 1.5 mm of the cyanobacterial mat show this cyanobacterial-purple bacterial association at depths of 300-1200 micrometers, corresponding to the zone below that of maximal oxygenic photosynthesis. Sharp gradients of oxygen and sulfide are established during the day at this microzone in the two cyanobacterial mats studied. The close association, the distribution pattern of this association and preliminary physiological experiments suggest a co-metabolism of sulfur by the two-membered community. This probable new genus of purple bacteria may also grow photoheterotrophically using organic carbon excreted by the cyanobacterium. Since the chemical gradients in the entire photic zone fluctuate widely in a diurnal cycle, both types of metabolism probably take place. During the morning and afternoon, sulfide migrates up to the photic zone allowing photoautotrophic metabolism with sulfide as the electron donor. During the day the photic zone is highly oxygenated and the purple bacteria may either use oxidized species of sulfur such as elemental sulfur and thiosulfate in the photoautotrophic mode or grow photoheterotrophically using organic carbon excreted by M. chthonoplastes. The new type of filamentous purple sulfur

Quantum dot-linked immunosorbent assay (QLISA) using orientation-directed antibodies.

Science.gov (United States)

Suzuki, Miho; Udaka, Hikari; Fukuda, Takeshi

2017-09-05

An approach similar to the enzyme-linked immunosorbent assay (ELISA), with the advantage of saving time and effort but exhibiting high performance, was developed using orientation-directed half-part antibodies immobilized on CdSe/ZnS quantum dots. ELISA is a widely accepted assay used to detect the presence of a target substance. However, it takes time to quantify the target with specificity and sensitivity owing to signal amplification. In this study, CdSe/ZnS quantum dots are introduced as bright and photobleaching-tolerant fluorescent materials. Since hydrophilic surface coating of quantum dots rendered biocompatibility and functional groups for chemical reactions, the quantum dots were modified with half-sized antibodies after partial reduction. The half-sized antibody could be bound to a quantum dot through a unique thiol site to properly display the recognition domain for the core process of ELISA, which is an antigen-antibody interaction. The reducing conditions were investigated to generate efficient conjugates of quantum dots and half-sized antibodies. This was applied to IL-6 detection, as the quantification of IL-6 is significant owing to its close relationships with various biomedical phenomena that cause different diseases. An ELISA-like assay with CdSe/ZnS quantum dot institution (QLISA; Quantum dot-linked immunosorbent assay) was developed to detect 0.05ng/mL IL-6, which makes it sufficiently sensitive as an immunosorbent assay. Copyright © 2017 Elsevier B.V. All rights reserved.

Characterization of antibacterial and adhesion properties of chitosan-modified glass ionomer cement.

Science.gov (United States)

Ibrahim, Marrwa A; Neo, Jennifer; Esguerra, Roxanna J; Fawzy, Amr S

2015-10-01

The aim is to investigate the effect of modifying the liquid phase of a conventional glass ionomer restorative material with different chitosan volume contents on the antibacterial properties and adhesion to dentin. The liquids of commercially available restorative glass ionomer cements (GIC) were modified with chitosan (CH) solutions at different volume contents (5%, 10%, 25%, and 50%). The GIC powders were mixed with the unmodified and the CH-modified liquids at the desired powder/liquid (P/L) ratio. For the characterization of the antibacterial properties, Streptococcus mutans biofilms were formed on GIC discs and characterized by scanning electron microscope (SEM), confocal microscopy, colony forming unit (CFU) count, and cell viability assay (MTS). The unmodified and CH-modified GICs were bonded to dentin surfaces and the micro-tensile bond strength (µTBs) was evaluated and the interface was investigated by SEM. Modification with CH solutions enhanced the antibacterial properties against S. mutans in terms of resistance to biofilm formation, CFU count, and MTS assay. Generally, significant improvement in the antibacterial properties was found with the increase in the CH volume content. Modification with 25% and 50% CH adversely affected the µTBs with predominant cohesive failure in the GIC. However, no difference was found between the control and the 5% and 10% CH-modified specimens. Incorporation of acidic solutions of chitosan in the polyacrylic acid liquid of GIC at v/v ratios of 5-10% improved the antibacterial properties of conventional glass ionomer cement against S. mutans without adversely affecting its bonding to dentin surface. © The Author(s) 2015.

Validation and modification of dried blood spot-based glycosylated hemoglobin assay for the longitudinal aging study in India.

Science.gov (United States)

Hu, Peifeng; Edenfield, Michael; Potter, Alan; Kale, Varsha; Risbud, Arun; Williams, Sharon; Lee, Jinkook; Bloom, David E; Crimmins, Eileen; Seeman, Teresa

2015-01-01

This study aims to validate a modified dried blood spot (DBS)-based glycosylated hemoglobin (HbA1c) assay protocol, after a pretest in India showed poor correlation between the original DBS-based protocol and venous results. The original protocol was tested on different chemistry analyzers and then simplified at the University of Washington (UW). A second pretest was conducted in India to validate the modified assay protocol, using 44 quality control specimens. Data from UW indicated that, using the original protocol, the correlation coefficients between DBS and venous results were above 0.98 on both Bio-Rad and Olympus chemistry analyzers. The protocol worked equally well on filter paper, with or without pre-treatment, and when the recommended amount of blood spot material, or less, was used. A second pretest of the modified protocol confirmed that DBS-based levels from both Olympus and Roche chemistry analyzers were well correlated with DBS results from UW (correlation coefficients were above 0.96), as well as with venous values (correlation coefficients were above 0.94). The DBS-based HbA1c values are highly correlated with venous results. The pre-treatment of filter paper does not appear to be necessary. The poor results from the first pretest are probably due to factors unrelated to the protocol, such as problems with the chemistry analyzer or assay reagents. © 2015 Wiley Periodicals, Inc.

Inter-laboratory validation of the modified murine local lymph node assay based on 5-bromo-2'-deoxyuridine incorporation.

Science.gov (United States)

Kojima, Hajime; Takeyoshi, Masahiro; Sozu, Takashi; Awogi, Takumi; Arima, Kazunori; Idehara, Kenji; Ikarashi, Yoshiaki; Kanazawa, Yukiko; Maki, Eiji; Omori, Takashi; Yuasa, Atsuko; Yoshimura, Isao

2011-01-01

The murine local lymph node assay (LLNA) is a well-established alternative to the guinea pig maximization test (GPMT) or Buehler test (BT) for the assessment of the skin sensitizing ability of a drug, cosmetic material, pesticide or industrial chemical. Instead of radioisotope using in this method, Takeyoshi M. et al. (2001) has developed a modified LLNA based on the 5-bromo-2'-deoxyuridine (BrdU) incorporation (LLNA:BrdU-ELISA). The LLNA:BrdU-ELISA is practically identical to the LLNA methodology excluding the use of BrdU, for which a single intraperitoneal injection of BrdU is made on day 4, and colorimetric detection of cell turnover. We conducted the validation study to evaluate the reliability and relevance of LLNA:BrdU-ELISA. The experiment involved 7 laboratories, wherein 10 chemicals were examined under blinded conditions. In this study, 3 chemicals were examined in all laboratories and the remaining 7 were examined in 3 laboratories. The data were expressed as the BrdU incorporation using an ELISA method for each group, and the stimulation index (SI) for each chemical-treated group was determined as the increase in the BrdU incorporation relative to the concurrent vehicle control group. An SI of 2 was set as the cut-off value for exhibiting skin sensitization activity. The results obtained in the experiments conducted for all 10 chemicals were sufficiently consistent with small variations in their SI values. The sensitivity, specificity, and accuracy of LLNA:BrdU-ELISA against those of GPMT/BT were 7/7 (100%), 3/3 (100%), and 10/10 (100%), respectively. Copyright © 2010 John Wiley & Sons, Ltd.

A radiolabel-release microwell assay for proteolytic enzymes present in cell culture media

International Nuclear Information System (INIS)

Rucklidge, G.J.; Milne, G.

1990-01-01

A modified method for the measurement of proteolytic enzyme activity in cell culture-conditioned media has been developed. Using the release of 3H-labeled peptides from 3H-labeled gelatin the method is performed in microwell plates. The substrate is insolubilized and attached to the wells by glutaraldehyde treatment, thus eliminating the need for a precipitation step at the end of the assay. The assay is sensitive, reproducible, and convenient for small sample volumes. The effect of different protease inhibitors on activity can be assessed rapidly allowing an early characterization of the enzyme. It can also be adapted to microplate spectrophotometric analysis by staining residual substrate with Coomassie blue

A protein chip membrane-capture assay for botulinum neurotoxin activity

International Nuclear Information System (INIS)

Marconi, Severine; Ferracci, Geraldine; Berthomieu, Maelys; Kozaki, Shunji; Miquelis, Raymond; Boucraut, Jose; Seagar, Michael

2008-01-01

Botulinum neurotoxins A and B (BoNT/A and B) are neuromuscular blocking agents which inhibit neurotransmission by cleaving the intra-cellular presynaptic SNARE proteins SNAP-25 and VAMP2, localized respectively in plasma membrane and synaptic vesicles. These neurotoxins are both dangerous pathogens and powerful therapeutic agents with numerous clinical and cosmetic applications. Consequently there is a need for in vitro assays of their biological activity to screen for potential inhibitors and to replace the widely used in vivo mouse assay. Surface plasmon resonance (SPR) was used to measure membrane vesicle capture by antibodies against SNAP-25 and VAMP2. Substrate cleavage by BoNTs modified capture providing a method to assay toxin activity. Firstly using synaptic vesicles as a substrate, a comparison of the EC 50 s for BoNT/B obtained by SPR, ELISA or flow cytometry indicated similar sensitivity although SPR assays were more rapid. Sonication of brain or neuronal cultures generated plasma membrane fragments with accessible intra-cellular epitopes adapted to measurement of BoNT/A activity. SPR responses were proportional to antigen concentration permitting detection of as little as 4 pM SNAP-25 in crude lysates. BoNT/A activity was assayed using monoclonal antibodies that specifically recognize a SNAP-25 epitope generated by the proteolytic action of the toxin. Incubation of intact primary cultured neurons with BoNT/A yielded an EC 50 of 0.5 pM. The SPR biosensor method was sensitive enough to monitor BoNT/A and B activity in cells cultured in a 96-well format providing an alternative to experimental animals for toxicological assays

Biomimetic synthesis of modified calcium phosphate fine powders and their in vitro studies

Energy Technology Data Exchange (ETDEWEB)

Gergulova, R., E-mail: rumigg@yahoo.com; Tepavitcharova, S., E-mail: rumigg@yahoo.com; Rabadjieva, D., E-mail: rumigg@yahoo.com; Sezanova, K., E-mail: rumigg@yahoo.com; Ilieva, R., E-mail: rumigg@yahoo.com [Institute of General and Inorganic Chemistry, Bulgarian Academy of Sciences, Acad. G. Bonchev Str., Bl. 11, 1113 Sofia (Bulgaria); Alexandrova, R.; Andonova-Lilova, B. [Institute of Experimental Morphology, Pathology and Anthropology with Museum, BAS, Acad. G. Bonchev Str., Bl. 25, Sofia (Bulgaria)

2013-12-16

Biomimetic approach and subsequent high-temperature treatment were used to synthesize ion modified calcium phosphate fine powders. Thus, using Simulated Body Fluid (SBF) as an ion modifier, a bi-phase mixture of ion modified β-tricalcium phosphate and hydroxyapatite (β-TCP + HA) was prepared. The use of SBF electrolyte solution enriched with Mg{sup 2+} or Zn{sup 2+} yielded monophase β-tricalcium phosphate additionally modified with Mg{sup 2+} or Zn{sup 2+} (Mg-β-TCP or Zn-β-TCP). The in vitro behavior of the prepared powders on cell viability and proliferation of murine BALB/c 3T3 fibroblasts and of human Lep 3 cells was studied by MTT test assays and Mosmann method after 72 h incubation. The relative cell viability was calculated.

Pertussis serology: assessment of IgG anti-PT ELISA for replacement of the CHO cell assay*

Science.gov (United States)

DALBY, TINE; SØRENSEN, CHARLOTTE; PETERSEN, JESPER WESTPHAL; KROGFELT, KAREN ANGELIKI

2010-01-01

Dalby T, Sørensen C, Petersen JW, Krogfelt KA. Pertussis serology: assessment of IgG anti-PT ELISA for replacement of the CHO cell assay. APMIS 2010; 118: 968–72. Two types of serological assays are commonly used for the assessment of pertussis vaccine-induced antibodies; the Chinese hamster ovary cell (CHO cell) assay and the immunoglobulin G (IgG) anti pertussis toxin (PT) enzyme-linked immunosorbent assay (IgG anti-PT ELISA). Recently, both the techniques have been modified to improve performance with sera with interfering activity (CHO cell assay) or with heat-treated sera (IgG anti-PT ELISA). These two improved techniques were compared by the analysis of 100 individual serum samples from a previous clinical trial and 213 sera from a longitudinal serum collection from 20 Danish adults recently vaccinated with the Danish acellular pertussis vaccine. The comparison showed a significant linear correlation between the results of the two assays with a p-value of ELISA can be used as a replacement for the often troublesome and time-consuming CHO cell assay for the measurement of vaccine-induced human antibodies to PT. PMID:21091778

Assaying Cellular Viability Using the Neutral Red Uptake Assay.

Science.gov (United States)

Ates, Gamze; Vanhaecke, Tamara; Rogiers, Vera; Rodrigues, Robim M

2017-01-01

The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration-dependent reduction of the uptake of neutral red after exposure to the xenobiotic under investigation. The neutral red uptake assay is mainly used for hazard assessment in in vitro toxicology applications. This method has also been introduced in regulatory recommendations as part of 3T3-NRU-phototoxicity-assay, which was regulatory accepted in all EU member states in 2000 and in the OECD member states in 2004 as a test guideline (TG 432). The present protocol describes the neutral red uptake assay using the human hepatoma cell line HepG2, which is often employed as an alternative in vitro model for human hepatocytes. As an example, the cytotoxicity of acetaminophen and acetyl salicylic acid is assessed.

Improved assay for measuring heparin binding to bull sperm

International Nuclear Information System (INIS)

Miller, D.J.; Ax, R.L.

1988-01-01

The binding of heparin to sperm has been used to study capacitation and to rank relative fertility of bulls. Previous binding assays were laborious, used 10 7 sperm per assay point, and required large amounts of radiolabeled heparin. A modified heparin-binding assay is described that used only 5 x 10 4 cells per incubation well and required reduced amounts of [ 3 H] heparin. The assay was performed in 96-well Millititer plates, enabling easy incubation and filtering. Dissociation constants and concentrations of binding sites did not differ if analyzed by Scatchard plots, Woolf plots, or by log-logit transformed weighted nonlinear least squares regression, except in the case of outliers. In such cases, Scatchard analysis was more sensitive to outliers. Nonspecific binding was insignificant using nonlinear logistic fit regression and a proportion graph. The effects were tested of multiple free-thawing of sperm in either a commercial egg yolk extender, 40 mM Tris buffer with 8% glycerol, or 40 mM Tris buffer without glycerol. Freeze-thawing in extender did not affect the dissociation constant or the concentration of binding sites. However, freeze-thawing three times in 40 mM Tris reduced the concentration of binding sites and lowered the dissociation constant (raised the affinity). The inclusion of glycerol in the 40 mM Tris did not significantly affect the estimated dissociation constant or the concentration of binding sites as compared to 40 mM Tris without glycerol

Comparison among manual instruments and PathFile and Mtwo rotary instruments to create a glide path in the root canal preparation of curved canals.

Science.gov (United States)

Alves, Vanessa de Oliveira; Bueno, Carlos Eduardo da Silveira; Cunha, Rodrigo Sanches; Pinheiro, Sérgio Luiz; Fontana, Carlos Eduardo; de Martin, Alexandre Sigrist

2012-01-01

Nickel-titanium rotary instruments reduce procedural errors and the time required to finish root canal preparation. The goal of this study was to evaluate the occurrences of apical transportation and canal aberrations produced with different instruments used to create a glide path in the preparation of curved root canals, namely manual K-files (Dentsply Maillefer, Ballaigues, Switzerland) and PathFile (Dentsply Maillefer) and Mtwo (Sweden and Martina, Padua, Italy) nickel-titanium rotary files. The mesial canals of 45 mandibular first and second molars (with curvature angles between 25° and 35°) were selected for this study. The specimens were divided randomly into 3 groups with 15 canals each, and canal preparation was performed by an endodontist using #10-15-20 K-type stainless steel manual files (group M), #13-16-19 PathFile rotary instruments (group PF), and #10-15-20 Mtwo rotary instruments (group MT). The double digital radiograph technique was used, pre- and postinstrumentation, to assess whether apical transportation and/or aberration in root canal morphology occurred. The initial and final images of the central axis of the canals were compared by superimposition through computerized analysis and with the aid of magnification. The specimens were analyzed by 3 evaluators, whose calibration was checked using the Kendall agreement test. No apical transportation or aberration in root canal morphology occurred in any of the teeth; therefore, no statistical analysis was conducted. Neither the manual instruments nor the PathFile or Mtwo rotary instruments used to create a glide path had any influence on the occurrence of apical transportation or produced any canal aberration. Copyright © 2012 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Evaluation of a modified IRMA for anti-D quantitation, using 3H protein A

International Nuclear Information System (INIS)

Dumasia, A.; Gupte, S.

1993-01-01

A modified immunoradiometric assay (IRMA) using tritiated ( 3 H) protein A was developed to estimate anti-D concentration. The main advantages of the assay were longer shelf life of the labelled reagent (more than two years); minimum radiation hazard and; low non specific binding. Levels of anti-D were estimated in 23 Rh (D) immunized women. A good correlation of anti-D concentration (μg/ml) with Rh antibody titre was observed (r=+ 0.89, P 3 H protein A IRMA correlated well with the severity of Rh-HDN. This assay could quantitate anti-D in sera having exclusively IgG 3 subtype. (author). 20 refs., 2 figs., 2 tabs

Factors Affecting Accuracy and Time Requirements of a Glucose Oxidase-Peroxidase Assay for Determination of Glucose

Science.gov (United States)

Accurate and rapid assays for glucose are desirable for analysis of glucose and starch in food and feedstuffs. An established colorimetric glucose oxidase-peroxidase method for glucose was modified to reduce analysis time, and evaluated for factors that affected accuracy. Time required to perform t...

GMO detection in food and feed through screening by visual loop-mediated isothermal amplification assays.

Science.gov (United States)

Wang, Cong; Li, Rong; Quan, Sheng; Shen, Ping; Zhang, Dabing; Shi, Jianxin; Yang, Litao

2015-06-01

Isothermal DNA/RNA amplification techniques are the primary methodology for developing on-spot rapid nucleic acid amplification assays, and the loop-mediated isothermal amplification (LAMP) technique has been developed and applied in the detection of foodborne pathogens, plant/animal viruses, and genetically modified (GM) food/feed contents. In this study, one set of LAMP assays targeting on eight frequently used universal elements, marker genes, and exogenous target genes, such as CaMV35S promoter, FMV35S promoter, NOS, bar, cry1Ac, CP4 epsps, pat, and NptII, were developed for visual screening of GM contents in plant-derived food samples with high efficiency and accuracy. For these eight LAMP assays, their specificity was evaluated by testing commercial GM plant events and their limits of detection were also determined, which are 10 haploid genome equivalents (HGE) for FMV35S promoter, cry1Ac, and pat assays, as well as five HGE for CaMV35S promoter, bar, NOS terminator, CP4 epsps, and NptII assays. The screening applicability of these LAMP assays was further validated successfully using practical canola, soybean, and maize samples. The results suggested that the established visual LAMP assays are applicable and cost-effective for GM screening in plant-derived food samples.

Development of a replication-competent lentivirus assay for dendritic cell-targeting lentiviral vectors

Directory of Open Access Journals (Sweden)

Daniel C Farley

Full Text Available It is a current regulatory requirement to demonstrate absence of detectable replication-competent lentivirus (RCL in lentiviral vector products prior to use in clinical trials. Immune Design previously described an HIV-1-based integration-deficient lentiviral vector for use in cancer immunotherapy (VP02. VP02 is enveloped with E1001, a modified Sindbis virus glycoprotein which targets dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN expressed on dendritic cells in vivo. Vector enveloped with E1001 does not transduce T-cell lines used in standard HIV-1-based RCL assays, making current RCL testing formats unsuitable for testing VP02. We therefore developed a novel assay to test for RCL in clinical lots of VP02. This assay, which utilizes a murine leukemia positive control virus and a 293F cell line expressing the E1001 receptor DC-SIGN, meets a series of evaluation criteria defined in collaboration with US regulatory authorities and demonstrates the ability of the assay format to amplify and detect a hypothetical RCL derived from VP02 vector components. This assay was qualified and used to test six independent GMP production lots of VP02, in which no RCL was detected. We propose that the evaluation criteria used to rationally design this novel method should be considered when developing an RCL assay for any lentiviral vector.

Detection of proteins using a colorimetric bio-barcode assay.

Science.gov (United States)

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

2007-01-01

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

Molecular motor transport through hollow nanowires

DEFF Research Database (Denmark)

Lard, Mercy; Ten Siethoff, Lasse; Generosi, Johanna

2014-01-01

-driven motion of fluorescent probes (actin filaments) through 80 nm wide, Al2O 3 hollow nanowires of micrometer length. The motor-driven transport is orders of magnitude faster than would be possible by passive diffusion. The system represents a necessary element for advanced devices based on gliding assays...

Random assay in radioimmunoassay: Feasibility and application compared with batch assay

Energy Technology Data Exchange (ETDEWEB)

Lee, Jung Min; Lee, Hwan Hee; Park, Sohyun; Kim, Tae Sung; Kim, Seok Ki [Dept. of Nuclear MedicineNational Cancer Center, Goyang (Korea, Republic of)

2016-12-15

The batch assay has been conventionally used for radioimmunoassay (RIA) because of its technical robustness and practical convenience. However, it has limitations in terms of the relative lag of report time due to the necessity of multiple assays in a small number of samples compared with the random assay technique. In this study, we aimed to verify whether the random assay technique can be applied in RIA and is feasible in daily practice. The coefficients of variation (CVs) of eight standard curves within a single kit were calculated in a CA-125 immunoradiometric assay (IRMA) for the reference of the practically ideal CV of the CA-125 kit. Ten standard curves of 10 kits from 2 prospectively collected lots (pLot) and 85 standard curves of 85 kits from 3 retrospectively collected lots (Lot) were obtained. Additionally, the raw measurement data of both 170 control references and 1123 patients' sera were collected retrospectively between December 2015 and January 2016. A standard curve of the first kit of each lot was used as a master standard curve for a random assay. The CVs of inter-kits were analyzed in each lot, respectively. All raw measurements were normalized by decay and radioactivity. The CA-125 values from control samples and patients' sera were compared using the original batch assay and random assay. In standard curve analysis, the CVs of inter-kits in pLots and Lots were comparable to those within a single kit. The CVs from the random assay with normalization were similar to those from the batch assay in the control samples (CVs % of low/high concentration; Lot1 2.71/1.91, Lot2 2.35/1.83, Lot3 2.83/2.08 vs. Lot1 2.05/1.21, Lot2 1.66/1.48, Lot3 2.41/2.14). The ICCs between the batch assay and random assay using patients' sera were satisfactory (Lot1 1.00, Lot2 0.999, Lot3 1.00). The random assay technique could be successfully applied to the conventional CA-125 IRMA kits. The random assay showed strong agreement with the batch assay. The

Irradiation influence on the detection of genetic-modified soybeans

International Nuclear Information System (INIS)

Villavicencio, A.L.C.H.; Araujo, M.M.; Baldasso, J.G.; Aquino, S.; Konietzny, U.; Greiner, R.

2004-01-01

Three soybean varieties were analyzed to evaluate the irradiation influence on the detection of genetic modification. Samples were treated in a 60 Co facility at dose levels of 0, 500, 800, and 1000 Gy. The seeds were at first analyzed by Comet Assay as a rapid screening irradiation detection method. Secondly, germination test was performed to detect the viability of irradiated soybeans. Finally, because of its high sensitivity, its specificity and rapidity the polimerase chain reaction was the method applied for genetic modified organism detection. The analysis of DNA by the single technique of microgel electrophoresis of single cells (DNA Comet Assay) showed that DNA damage increased with increasing radiation doses. No negative influence of irradiation on the genetic modification detection was found

Effects of Surface Epitope Coverage on the Sensitivity of Displacement Assays that Employ Modified Nanoparticles: Using Bisphenol A as a Model Analyte

Directory of Open Access Journals (Sweden)

Yang Lu

2016-08-01

Full Text Available With the ever-increasing use of nanoparticles in immunosensors, a fundamental study on the effect of epitope density is presented herein, with a small molecule epitope, on the performance of the displacement assay format in an enzyme-linked immunosorbent assay (ELISA. Thiolated bisphenol A (BPA functionalized gold nanoparticles (cysBPAv-AuNPs and specific anti-BPA antibodies are employed for this purpose. It is shown that the displacement of cysBPAv-AuNPs bound to the immobilized antibodies was influenced by both the avidity of bound cysBPAv-AuNPs and the concentration of free BPA to displace it. The importance of surface epitope density was that it changed the number of epitopes in close proximity to the antibody-binding site. This then influenced the avidity of cysBPAv-AuNPs bound to the immobilized antibody. Furthermore, the molar epitope concentration in an assay appears to affect the degree of antibody binding site saturation. Controlling surface epitope density of the functionalized nanoparticles and molar epitope concentration in an assay leads to a decrease of the concentration of free BPA required to displace the bound cysBPAv-AuNP, and hence better assay performance with regards to the D50 value and dynamic range in the displacement assay.

Solid solution hardening in face centered binary alloys: Gliding statistics of a dislocation in random solid solution by atomistic simulation

International Nuclear Information System (INIS)

Patinet, S.

2009-12-01

The glide of edge and screw dislocation in solid solution is modeled through atomistic simulations in two model alloys of Ni(Al) and Al(Mg) described within the embedded atom method. Our approach is based on the study of the elementary interaction between dislocations and solutes to derive solid solution hardening of face centered cubic binary alloys. We identify the physical origins of the intensity and range of the interaction between a dislocation and a solute atom. The thermally activated crossing of a solute atom by a dislocation is studied at the atomistic scale. We show that hardening of edge and screw segments are similar. We develop a line tension model that reproduces quantitatively the atomistic calculations of the flow stress. We identify the universality class to which the dislocation depinning transition in solid solution belongs. (author)

Use of toxicity assays for evaluating the effectiveness of groundwater remediation with Fenton’s reagent

DEFF Research Database (Denmark)

Kusk, Kresten Ole; Bennedsen, Lars; Christophersen, Mette

2011-01-01

evaluates in situ chemical oxidation (ISCO) using modified Fenton’s reagent (H2O2 + chelated Fe2+) as a groundwater remedy. Three injections were performed over a period to test treatment efficacy. Performance monitoring samples were collected from two depths both prior to and during treatment, and analyzed...... treatment with Fenton’s reagent the toxicity had increased and now needed 7100 times dilution to reduce toxicity to the LC10 probably due to mobilization of metals. It is concluded that toxicity assay is a useful tool for evaluating samples from contaminated sites and that toxicity assays and chemical...

Porphyromonas gingivalis and related bacteria: from colonial pigmentation to the type IX secretion system and gliding motility

Science.gov (United States)

Nakayama, K

2015-01-01

Porphyromonas gingivalis is a gram-negative, non-motile, anaerobic bacterium implicated as a major pathogen in periodontal disease. P. gingivalis grows as black-pigmented colonies on blood agar, and many bacteriologists have shown interest in this property. Studies of colonial pigmentation have revealed a number of important findings, including an association with the highly active extracellular and surface proteinases called gingipains that are found in P. gingivalis. The Por secretion system, a novel type IX secretion system (T9SS), has been implicated in gingipain secretion in studies using non-pigmented mutants. In addition, many potent virulence proteins, including the metallocarboxypeptidase CPG70, 35 kDa hemin-binding protein HBP35, peptidylarginine deiminase PAD and Lys-specific serine endopeptidase PepK, are secreted through the T9SS. These findings have not been limited to P. gingivalis but have been extended to other bacteria belonging to the phylum Bacteroidetes. Many Bacteroidetes species possess the T9SS, which is associated with gliding motility for some of these bacteria. PMID:25546073

Matrix effects of TRU [transuranic] assays using the SWEPP PAN assay system

International Nuclear Information System (INIS)

Smith, J.R.

1990-08-01

The Drum Assay System (DAS) at the Stored Waste Experimental Pilot Plant (SWEPP) is a second-generation active-passive neutron assay system. It has been used to assay over 5000 208-liter drums of transuranic waste from the Rocky Flats Plant (RFP). Data from these assays have been examined and compared with the assays performed at Rocky Flats, mainly utilize counting of 239 Pu gamma rays. For the most part the passive assays are in very good agreement with the Rocky Flats assays. The active assays are strongly correlated with the results of the other two methods, but require matrix-dependent correction factors beyond those provided by the system itself. A set of matrix-dependent correction factors has been developed from the study of the assay results. 3 refs., 4 figs., 3 tabs

Collective behavior of minus-ended motors in mitotic microtubule asters gliding toward DNA

International Nuclear Information System (INIS)

Athale, Chaitanya A; Dinarina, Ana; Nedelec, Francois; Karsenti, Eric

2014-01-01

Microtubules (MTs) nucleated by centrosomes form star-shaped structures referred to as asters. Aster motility and dynamics is vital for genome stability, cell division, polarization and differentiation. Asters move either toward the cell center or away from it. Here, we focus on the centering mechanism in a membrane independent system of Xenopus cytoplasmic egg extracts. Using live microscopy and single particle tracking, we find that asters move toward chromatinized DNA structures. The velocity and directionality profiles suggest a random-walk with drift directed toward DNA. We have developed a theoretical model that can explain this movement as a result of a gradient of MT length dynamics and MT gliding on immobilized dynein motors. In simulations, the antagonistic action of the motor species on the radial array of MTs leads to a tug-of-war purely due to geometric considerations and aster motility resembles a directed random-walk. Additionally, our model predicts that aster velocities do not change greatly with varying initial distance from DNA. The movement of asymmetric asters becomes increasingly super-diffusive with increasing motor density, but for symmetric asters it becomes less super-diffusive. The transition of symmetric asters from superdiffusive to diffusive mobility is the result of number fluctuations in bound motors in the tug-of-war. Overall, our model is in good agreement with experimental data in Xenopus cytoplasmic extracts and predicts novel features of the collective effects of motor-MT interactions. (paper)

Development and validation of a quantitative PCR assay for Ichthyophonus spp.

Science.gov (United States)

White, Vanessa C; Morado, J Frank; Crosson, Lisa M; Vadopalas, Brent; Friedman, Carolyn S

2013-04-29

Members of the genus Ichthyophonus are trophically transmitted, cosmopolitan parasites that affect numerous fish species worldwide. A quantitative PCR (qPCR) assay specific for genus Ichthyophonus 18S ribosomal DNA was developed for parasite detection and surveillance. The new assay was tested for precision, repeatability, reproducibility, and both analytical sensitivity and specificity. Diagnostic sensitivity and specificity were estimated using tissue samples from a wild population of walleye pollock Theragra chalcogramma. Ichthyophonus sp. presence in tissue samples was determined by qPCR, conventional PCR (cPCR), and histology. Parasite prevalence estimates varied depending upon the detection method employed and tissue type tested. qPCR identified the greatest number of Ichthyophonus sp.-positive cases when applied to walleye pollock skeletal muscle. The qPCR assay proved sensitive and specific for Ichthyophonus spp. DNA, but like cPCR, is only a proxy for infection. When compared to cPCR, qPCR possesses added benefits of parasite DNA quantification and a 100-fold increase in analytical sensitivity. Because this novel assay is specific for known members of the genus, it is likely appropriate for detecting Ichthyophonus spp. DNA in various hosts from multiple regions. However, species-level identification and isotype variability would require DNA sequencing. In addition to distribution and prevalence applications, this assay could be modified and adapted for use with zooplankton or environmental samples. Such applications could aid in investigating alternate routes of transmission and life history strategies typical to members of the genus Ichthyophonus.

Production of recombinant proteins from Plasmodium falciparum in Escherichia coli.

Science.gov (United States)

Guerra, Ángela Patricia; Calvo, Eliana Patricia; Wasserman, Moisés; Chaparro-Olaya, Jacqueline

2016-02-23

The production of recombinant proteins is essential for the characterization and functional study of proteins from Plasmodium falciparum. However, the proteins of P. falciparum are among the most challenging to express, and when expression is achieved, the recombinant proteins usually fold incorrectly and lead to the formation of inclusion bodies.  To obtain and purify four recombinant proteins and to use them as antigens to produce polyclonal antibodies. The production efficiency and solubility were evaluated as the proteins were expressed in two genetically modified strains of Escherichia coli to favor the production of heterologous proteins (BL21-CodonPlus (DE3)-RIL and BL21-pG-KJE8).  The four recombinant P. falciparum proteins corresponding to partial sequences of PfMyoA (Myosin A) and PfGAP50 (gliding associated protein 50), and the complete sequences of PfMTIP (myosin tail interacting protein) and PfGAP45 (gliding associated protein 45), were produced as glutathione S-transferase-fusion proteins, purified and used for immunizing mice.  The protein expression was much more efficient in BL21-CodonPlus, the strain that contains tRNAs that are rare in wild-type E. coli, compared to the expression in BL21-pG-KJE8. In spite of the fact that BL21-pG-KJE8 overexpresses chaperones, this strain did not minimize the formation of inclusion bodies.  The use of genetically modified strains of E. coli was essential to achieve high expression levels of the four evaluated P. falciparum proteins and lead to improved solubility of two of them. The approach used here allowed us to obtain and purify four P. falciparum proteins in enough quantity to produce polyclonal antibodies in mice, and a fair amount of two pure and soluble recombinant proteins for future assays.

Comparison of Batch Assay and Random Assay Using Automatic Dispenser in Radioimmunoassay

Energy Technology Data Exchange (ETDEWEB)

Moon, Seung Hwan; Jang, Su Jin; Kang, Ji Yeon; Lee, Dong Soo; Chung, June Key; Lee, Myung Chul [Seoul Metropolitan Government Seoul National University Boramae Medical Center, Seoul (Korea, Republic of); Lee, Ho Young; Shin, Sun Young; Min, Gyeong Sun; Lee, Hyun Joo [Seoul National University college of Medicine, Seoul (Korea, Republic of)

2009-08-15

Radioimmunoassay (RIA) was usually performed by the batch assay. To improve the efficiency of RIA without increase of the cost and time, random assay could be a choice. We investigated the possibility of the random assay using automatic dispenser by assessing the agreement between batch assay and random assay. The experiments were performed with four items; Triiodothyronine (T3), free thyroxine (fT4), Prostate specific antigen (PSA), Carcinoembryonic antigen (CEA). In each item, the sera of twenty patients, the standard, and the control samples were used. The measurements were done 4 times with 3 hour time intervals by random assay and batch assay. The coefficient of variation (CV) of the standard samples and patients' data in T3, fT4, PSA, and CEA were assessed. ICC (Intraclass correlation coefficient) and coefficient of correlation were measured to assessing the agreement between two methods. The CVs (%) of T3, fT4, PSA, and CEA measured by batch assay were 3.2+-1.7%, 3.9+-2.1%, 7.1+-6.2%, 11.2+-7.2%. The CVs by random assay were 2.1+-1.7%, 4.8+-3.1%, 3.6+-4.8%, and 7.4+-6.2%. The ICC between the batch assay and random assay were 0.9968 (T3), 0.9973 (fT4), 0.9996 (PSA), and 0.9901 (CEA). The coefficient of correlation between the batch assay and random assay were 0.9924(T3), 0.9974 (fT4), 0.9994 (PSA), and 0.9989 (CEA) (p<0.05). The results of random assay showed strong agreement with the batch assay in a day. These results suggest that random assay using automatic dispenser could be used in radioimmunoassay

Comparison of Batch Assay and Random Assay Using Automatic Dispenser in Radioimmunoassay

International Nuclear Information System (INIS)

Moon, Seung Hwan; Jang, Su Jin; Kang, Ji Yeon; Lee, Dong Soo; Chung, June Key; Lee, Myung Chul; Lee, Ho Young; Shin, Sun Young; Min, Gyeong Sun; Lee, Hyun Joo

2009-01-01

Radioimmunoassay (RIA) was usually performed by the batch assay. To improve the efficiency of RIA without increase of the cost and time, random assay could be a choice. We investigated the possibility of the random assay using automatic dispenser by assessing the agreement between batch assay and random assay. The experiments were performed with four items; Triiodothyronine (T3), free thyroxine (fT4), Prostate specific antigen (PSA), Carcinoembryonic antigen (CEA). In each item, the sera of twenty patients, the standard, and the control samples were used. The measurements were done 4 times with 3 hour time intervals by random assay and batch assay. The coefficient of variation (CV) of the standard samples and patients' data in T3, fT4, PSA, and CEA were assessed. ICC (Intraclass correlation coefficient) and coefficient of correlation were measured to assessing the agreement between two methods. The CVs (%) of T3, fT4, PSA, and CEA measured by batch assay were 3.2±1.7%, 3.9±2.1%, 7.1±6.2%, 11.2±7.2%. The CVs by random assay were 2.1±1.7%, 4.8±3.1%, 3.6±4.8%, and 7.4±6.2%. The ICC between the batch assay and random assay were 0.9968 (T3), 0.9973 (fT4), 0.9996 (PSA), and 0.9901 (CEA). The coefficient of correlation between the batch assay and random assay were 0.9924(T3), 0.9974 (fT4), 0.9994 (PSA), and 0.9989 (CEA) (p<0.05). The results of random assay showed strong agreement with the batch assay in a day. These results suggest that random assay using automatic dispenser could be used in radioimmunoassay

Monte carlo feasibility study of an active neutron assay technique for full-volume UF{sub 6} cylinder assay using a correlated interrogation source

Energy Technology Data Exchange (ETDEWEB)

Miller, Karen A., E-mail: kamiller@lanl.gov [Los Alamos National Laboratory, Los Alamos, P.O. Box 1663 MS E540, NM 87545 (United States); Menlove, Howard O.; Swinhoe, Martyn T.; Marlow, Johnna B. [Los Alamos National Laboratory, Los Alamos, P.O. Box 1663 MS E540, NM 87545 (United States)

2013-03-01

Uranium cylinder assay plays an important role in the nuclear material accounting at gas centrifuge enrichment plants. The Passive Neutron Enrichment Meter (PNEM) was designed to determine uranium mass and enrichment in 30B and 48Y cylinders using total neutron and coincidence counting in the passive mode. 30B and 48Y cylinders are used to hold bulk UF{sub 6} feed, product, and tails at enrichment plants. In this paper, we report the results of a Monte-Carlo-based feasibility study for an active uranium cylinder assay system based on the PNEM design. There are many advantages of the active technique such as a shortened count time and a more direct measure of {sup 235}U content. The active system is based on a modified PNEM design and uses a {sup 252}Cf source as the correlated, active interrogation source. We show through comparison with a random AmLi source of equal strength how the use of a correlated driver significantly boosts the active signal and reduces the statistical uncertainty. We also discuss ways in which an active uranium cylinder assay system can be optimized to minimize background from {sup 238}U fast-neutron induced fission and direct counts from the interrogation source.

Modified biomolecule as potential vehicle for buccal delivery of doxepin.

Science.gov (United States)

Laffleur, Flavia; Zilio, Martina; Shuwisitkul, Duangratana

2016-10-01

Doxepin is a traditional tricyclic antidepressant with analgesic and anesthetic properties when applied topically to the mucosa. Doxepin is one approach in treating insomnia and depression in Parkinson's disease. Patients with Parkinson's disease suffer difficulties in swallowing. Therefore, it was the aim of this study to develop a buccal-adhesive delivery system. Pectin was modified with cysteine. Stability assays in form of disintegration assay according to the Ph.Eur were performed. Furthermore, bioadhesiveness on buccal mucosa was investigated incorporating the drug doxepin. The adhesiveness was improved 1.4-fold and revealed a sustained release over 3 h. Taking these findings into account, the modifications render this designed excipient fruitful for buccal delivery.

Radioreceptor opioid assay

International Nuclear Information System (INIS)

Miller, R.J.; Chang, K.-J.

1981-01-01

A radioreceptor assay is described for assaying opioid drugs in biological fluids. The method enables the assay of total opioid activity, being specific for opioids as a class but lacking specificity within the class. A radio-iodinated opioid and the liquid test sample are incubated with an opiate receptor material. The percentage inhibition of the binding of the radio-iodinated compound to the opiate receptor is calculated and the opioid activity of the test liquid determined from a standard curve. Examples of preparing radio-iodinated opioids and assaying opioid activity are given. A test kit for the assay is described. Compared to other methods, this assay is cheap, easy and rapid. (U.K.)

Utilization of exploration-based learning and video-assisted learning to teach GlideScope videolaryngoscopy.

Science.gov (United States)

Johnston, Lindsay C; Auerbach, Marc; Kappus, Liana; Emerson, Beth; Zigmont, Jason; Sudikoff, Stephanie N

2014-01-01

GlideScope (GS) is used in pediatric endotracheal intubation (ETI) but requires a different technique compared to direct laryngoscopy (DL). This article was written to evaluate the efficacy of exploration-based learning on procedural performance using GS for ETI of simulated pediatric airways and establish baseline success rates and procedural duration using DL in airway trainers among pediatric providers at various levels. Fifty-five pediatric residents, fellows, and faculty from Pediatric Critical Care, NICU, and Pediatric Emergency Medicine were enrolled. Nine physicians from Pediatric Anesthesia benchmarked expert performance. Participants completed a demographic survey and viewed a video by the GS manufacturer. Subjects spent 15 minutes exploring GS equipment and practicing the intubation procedure. Participants then intubated neonatal, infant, child, and adult airway simulators, using GS and DL, in random order. Time to ETI was recorded. Procedural performance after exploration-based learning, measured as time to successful ETI, was shorter for DL than for GS for neonatal and child airways at the.05 significance level. Time to ETI in adult airway using DL was correlated with experience level (p =.01). Failure rates were not different among subgroups. A brief video and period of exploration-based learning is insufficient for implementing a new technology. Pediatricians at various levels of training intubated simulated airways faster using DL than GS.

Identification and quantification of genetically modified Moonshade carnation lines using conventional and TaqMan real-time polymerase chain reaction methods.

Science.gov (United States)

Li, Peng; Jia, Junwei; Bai, Lan; Pan, Aihu; Tang, Xueming

2013-07-01

Genetically modified carnation (Dianthus caryophyllus L.) Moonshade was approved for planting and commercialization in several countries from 2004. Developing methods for analyzing Moonshade is necessary for implementing genetically modified organism labeling regulations. In this study, the 5'-transgene integration sequence was isolated using thermal asymmetric interlaced (TAIL)-PCR. Based upon the 5'-transgene integration sequence, conventional and TaqMan real-time PCR assays were established. The relative limit of detection for the conventional PCR assay was 0.05 % for Moonshade using 100 ng total carnation genomic DNA, corresponding to approximately 79 copies of the carnation haploid genome, and the limits of detection and quantification of the TaqMan real-time PCR assay were estimated to be 51 and 254 copies of haploid carnation genomic DNA, respectively. These results are useful for identifying and quantifying Moonshade and its derivatives.

Screening of anionic-modified polymers in terms of stability, disintegration, and swelling behavior.

Science.gov (United States)

Laffleur, Flavia; Ijaz, Muhammad; Menzel, Claudia

2017-11-01

This study aimed to screen the stability, disintegration, and swelling behavior of chemically modified anionic polymers. Investigated polymers were well-known and widely used staples of the pharmaceutical and medical field, namely, alginate (AL), carboxymethyl cellulose (CMC), polycarbophil (PC), and hyaluronic acid (HA). On the basis of amide bond formation between the carboxylic acid moieties of anionic polymers and the primary amino group of the modification ligand cysteine (CYS), the modified polymers were obtained. Unmodified polymers served as controls throughout all studies. With the Ellman's assay, modification degrees were determined of synthesized polymeric excipients. Stability assay in terms of erosion study at physiological conditions were performed. Moreover, water uptake of compressed polymeric discs were evaluated and further disintegration studies according to the USP were carried out to define the potential ranking. Results ranking figured out PCCYS > CMCCYS > HACYS > ALCYS in terms of water uptake capacity compared to respective controls. Cell viability assays on Caco-2 cell line as well as on RPMI 2650 (ATTC CCL30) proved modification not being harmful to those. Due to the results of this study, an intense screening of prominent anionic polymer derivate was performed in order to help the pharmaceutical research for the best choice of polymeric excipients for developments of controlled drug release systems.

An efficient genotyping method for genome-modified animals and human cells generated with CRISPR/Cas9 system.

Science.gov (United States)

Zhu, Xiaoxiao; Xu, Yajie; Yu, Shanshan; Lu, Lu; Ding, Mingqin; Cheng, Jing; Song, Guoxu; Gao, Xing; Yao, Liangming; Fan, Dongdong; Meng, Shu; Zhang, Xuewen; Hu, Shengdi; Tian, Yong

2014-09-19

The rapid generation of various species and strains of laboratory animals using CRISPR/Cas9 technology has dramatically accelerated the interrogation of gene function in vivo. So far, the dominant approach for genotyping of genome-modified animals has been the T7E1 endonuclease cleavage assay. Here, we present a polyacrylamide gel electrophoresis-based (PAGE) method to genotype mice harboring different types of indel mutations. We developed 6 strains of genome-modified mice using CRISPR/Cas9 system, and utilized this approach to genotype mice from F0 to F2 generation, which included single and multiplexed genome-modified mice. We also determined the maximal detection sensitivity for detecting mosaic DNA using PAGE-based assay as 0.5%. We further applied PAGE-based genotyping approach to detect CRISPR/Cas9-mediated on- and off-target effect in human 293T and induced pluripotent stem cells (iPSCs). Thus, PAGE-based genotyping approach meets the rapidly increasing demand for genotyping of the fast-growing number of genome-modified animals and human cell lines created using CRISPR/Cas9 system or other nuclease systems such as TALEN or ZFN.

PCR-free quantitative detection of genetically modified organism from raw materials. An electrochemiluminescence-based bio bar code method.

Science.gov (United States)

Zhu, Debin; Tang, Yabing; Xing, Da; Chen, Wei R

2008-05-15

A bio bar code assay based on oligonucleotide-modified gold nanoparticles (Au-NPs) provides a PCR-free method for quantitative detection of nucleic acid targets. However, the current bio bar code assay requires lengthy experimental procedures including the preparation and release of bar code DNA probes from the target-nanoparticle complex and immobilization and hybridization of the probes for quantification. Herein, we report a novel PCR-free electrochemiluminescence (ECL)-based bio bar code assay for the quantitative detection of genetically modified organism (GMO) from raw materials. It consists of tris-(2,2'-bipyridyl) ruthenium (TBR)-labeled bar code DNA, nucleic acid hybridization using Au-NPs and biotin-labeled probes, and selective capture of the hybridization complex by streptavidin-coated paramagnetic beads. The detection of target DNA is realized by direct measurement of ECL emission of TBR. It can quantitatively detect target nucleic acids with high speed and sensitivity. This method can be used to quantitatively detect GMO fragments from real GMO products.

Transferability study of CHO cell clustering assays for monitoring of pertussis toxin activity in acellular pertussis vaccines.

Science.gov (United States)

Isbrucker, R; Daas, A; Wagner, L; Costanzo, A

2016-01-01

Current regulations for acellular pertussis (aP) vaccines require that they are tested for the presence of residual or reversion-derived pertussis toxin (PTx) activity using the mouse histamine sensitisation test (HIST). Although a CHO cell clustering assay can be used by manufacturers to verify if sufficient inactivation of the substance has occurred in-process, this assay cannot be used at present for the final product due to the presence of aluminium adjuvants which interfere with mammalian cell cultures. Recently, 2 modified CHO cell clustering assays which accommodate for the adjuvant effects have been proposed as alternatives to the HIST. These modified assays eliminate the adjuvant-induced cytotoxicity either through dilution of the vaccine (called the Direct Method) or by introducing a porous barrier between the adjuvant and the cells (the Indirect Method). Transferability and suitability of these methods for testing of products present on the European market were investigated during a collaborative study organised by the European Directorate for the Quality of Medicines & HealthCare (EDQM). Thirteen laboratories participated in this study which included 4 aP-containing vaccines spiked by addition of PTx. This study also assessed the transferability of a standardised CHO cell clustering assay protocol for use with non-adjuvanted PTx preparations. Results showed that the majority of laboratories were able to detect the PTx spike in all 4 vaccines at concentrations of 4 IU/mL or lower using the Indirect Method. This sensitivity is in the range of the theoretical sensitivity of the HIST. The Direct Method however did not show the expected results and would need additional development work.

Assay reproducibility in clinical studies of plasma miRNA.

Directory of Open Access Journals (Sweden)

Jonathan Rice

Full Text Available There are increasing reports of plasma miRNAs as biomarkers of human disease but few standards in methodologic reporting, leading to inconsistent data. We systematically reviewed plasma miRNA studies published between July 2013-June 2014 to assess methodology. Six parameters were investigated: time to plasma extraction, methods of RNA extraction, type of miRNA, quantification, cycle threshold (Ct setting, and methods of statistical analysis. We compared these data with a proposed standard methodologic technique. Beginning with initial screening for 380 miRNAs using microfluidic array technology and validation in an additional cohort of patients, we compared 11 miRNAs that exhibited differential expression between 16 patients with benign colorectal neoplasms (advanced adenomas and 16 patients without any neoplasm (controls. Plasma was isolated immediately, 12, 24, 48, or 72 h following phlebotomy. miRNA was extracted using two different techniques (Trizol LS with pre-amplification or modified miRNeasy. We performed Taqman-based RT-PCR assays for the 11 miRNAs with subsequent analyses using a variable Ct setting or a fixed Ct set at 0.01, 0.03, 0.05, or 0.5. Assays were performed in duplicate by two different operators. RNU6 was the internal reference. Systematic review yielded 74 manuscripts meeting inclusion criteria. One manuscript (1.4% documented all 6 methodological parameters, while < 5% of studies listed Ct setting. In our proposed standard technique, plasma extraction ≤12 h provided consistent ΔCt. miRNeasy extraction yielded higher miRNA concentrations and fewer non-expressed miRNAs compared to Trizol LS (1/704 miRNAs [0.14%] vs 109/704 miRNAs [15%], not expressed, respectively. A fixed Ct bar setting of 0.03 yielded the most reproducible data, provided that <10% miRNA were non-expressed. There was no significant intra-operator variability. There was significant inter-operator variation using Trizol LS extraction, while this was

Biobarcode assay for the oral anticoagulant acenocoumarol.

Science.gov (United States)

Broto, Marta; Salvador, J Pablo; Galve, Roger; Marco, M Pilar

2018-02-01

A novel approach for therapeutic drug monitoring of oral anticoagulants (OA) in clinical samples is reported, based on a NP-based biobarcode assay. The proposed strategy uses specific antibodies for acenocumarol (ACL) covalently bound to magnetic particles (pAb236-MP) and a bioconjugate competitor (hACL-BSA) linked to encoded polystyrene probes (hACL-BSA-ePSP) on a classical competitive immunochemical format. By using this scheme ACL can be detected in low nM range (LOD, 0.96 ± 0.26, N = 3, in buffer) even in complex samples such as serum or plasma (LOD 4 ± 1). The assay shows a high reproducibility (%CV 1.1 day-to-day) and is robust, as it is demonstrated by the fact that ACL can be quantified in complex biological samples with a very good accuracy (slope = 0.97 and R 2 = 0.91, of the linear regression obtained when analyzing spiked vs measured values). Moreover, we have demonstrated that the biobarcode approach has the potential to overcome one of the main challenges of the multiplexed diagnostic, which is the possibility to measure in a single run biomarker targets present at different concentration ranges. Thus, it has been proven that the signal and the detectability can be modulated by just modifying the oligonucleotide load of the encoded probes. This fact opens the door for combining in the same assay encoded probes with the necessary oligonucleotide load to achieve the detectability required for each biomarker target. Copyright © 2017 Elsevier B.V. All rights reserved.

New Insights into Butyrylcholinesterase Activity Assay: Serum Dilution Factor as a Crucial Parameter.

Directory of Open Access Journals (Sweden)

Joanna Jońca

Full Text Available Butyrylcholinesterase (BChE activity assay and inhibitor phenotyping can help to identify patients at risk of prolonged paralysis following the administration of neuromuscular blocking agents. The assay plays an important role in clinical chemistry as a good diagnostic marker for intoxication with pesticides and nerve agents. Furthermore, the assay is also commonly used for in vitro characterization of cholinesterases, their toxins and drugs. There is still lack of standardized procedure for measurement of BChE activity and many laboratories use different substrates at various concentrations. The purpose of this study was to validate the BChE activity assay to determine the best dilution of human serum and the most optimal concentration of substrates and inhibitors. Serum BChE activity was measured using modified Ellman's method applicable for a microplate reader. We present our experience and new insights into the protocol for high-throughput routine assays of human plasma cholinesterase activities adapted to a microplate reader. During our routine assays used for the determination of BChE activity, we have observed that serum dilution factor influences the results obtained. We show that a 400-fold dilution of serum and 5mM S-butyrylthiocholine iodide can be successfully used for the accurate measurement of BChE activity in human serum. We also discuss usage of various concentrations of dibucaine and fluoride in BChE phenotyping. This study indicates that some factors of such a multicomponent clinical material like serum can influence kinetic parameters of the BChE. The observed inhibitory effect is dependent on serum dilution factor used in the assay.

Modifiers of notch transcriptional activity identified by genome-wide RNAi

Directory of Open Access Journals (Sweden)

Firnhaber Christopher B

2010-10-01

Full Text Available Abstract Background The Notch signaling pathway regulates a diverse array of developmental processes, and aberrant Notch signaling can lead to diseases, including cancer. To obtain a more comprehensive understanding of the genetic network that integrates into Notch signaling, we performed a genome-wide RNAi screen in Drosophila cell culture to identify genes that modify Notch-dependent transcription. Results Employing complementary data analyses, we found 399 putative modifiers: 189 promoting and 210 antagonizing Notch activated transcription. These modifiers included several known Notch interactors, validating the robustness of the assay. Many novel modifiers were also identified, covering a range of cellular localizations from the extracellular matrix to the nucleus, as well as a large number of proteins with unknown function. Chromatin-modifying proteins represent a major class of genes identified, including histone deacetylase and demethylase complex components and other chromatin modifying, remodeling and replacement factors. A protein-protein interaction map of the Notch-dependent transcription modifiers revealed that a large number of the identified proteins interact physically with these core chromatin components. Conclusions The genome-wide RNAi screen identified many genes that can modulate Notch transcriptional output. A protein interaction map of the identified genes highlighted a network of chromatin-modifying enzymes and remodelers that regulate Notch transcription. Our results open new avenues to explore the mechanisms of Notch signal regulation and the integration of this pathway into diverse cellular processes.

The microculture tetrazolium assay (MTA): another colorimetric method of testing Plasmodium falciparum chemosensitivity.

Science.gov (United States)

Delhaes, L; Lazaro, J E; Gay, F; Thellier, M; Danis, M

1999-01-01

Malarial lactate dehydrogenase (LDH), which uses 3-acetyl pyridine adenine dinucleotide as coenzyme in a reaction leading to the formation of pyruvate from L-lactate, may be used to study the susceptibility of Plasmodium falciparum to a drug in vitro. Several methods to determine the activity of this enzyme are available. One, the colorimetric method of Makler and colleagues, was modified slightly, by using sodium-2,3-bis-[2-methoxy-4-nitro-5-sulphophenyl]-2H-tetrazolium-5 - carboxanilide (XTT) and following the reaction by measuring the optical density at 450 nm. Using two, culture-adapted strains of P. falciparum, this LDH assay was compared with the unmodified Makler's assay and with the isotopic microtest based on the incorporation of tritium-labelled hypoxanthine. Fresh, clinical P. falciparum isolates were also tested in the presence of several drugs, including chloroquine, mefloquine, quinine, halofantrine, atovaquone and qinghaosu derivatives. The results of the three assays were correlated for all the drugs tested except atovaquone. The two enzymatic assays are non-radioactive, rapid, reliable, inexpensive to perform and semi-automatic. However, they do require an initial parasitaemia of 2% with a haematocrit of 1.8%.

Evaluation of Antibacterial Activity of Titanium Surface Modified by PVD/PACVD Process.

Science.gov (United States)

Ji, Min-Kyung; Lee, Min-Joo; Park, Sang-Won; Lee, Kwangmin; Yun, Kwi-Dug; Kim, Hyun-Seung; Oh, Gye-Jeong; Kim, Ji-Hyun; Lim, Hyun-Pil

2016-02-01

The aim of this study was to evaluate the response of Streptococcus mutans (S. mutans) via crystal violet staining assay on titanium surface modified by physical vapor deposition/plasma assisted chemical vapor deposition process. Specimens were divided into the following three groups: polished titanium (control group), titanium modified by DC magnetron sputtering (group TiN-Ti), and titanium modified by plasma nitriding (group N-Ti). Surface characteristics of specimens were observed by using nanosurface 3D optical profiler and field emission scanning electron microscope. Group TiN-Ti showed TiN layer of 1.2 microm in thickness. Group N-Ti was identified as plasma nitriding with X-ray photoelectron spectroscopy. Roughness average (Ra) of all specimens had values 0.05). Within the process condition of this study, modified titanium surfaces by DC magnetron sputtering and plasma nitriding did not influence the adhesion of S. mutans.

Enhancement of sorption capacity of cocoa shell biomass modified with non-thermal plasma for removal of both cationic and anionic dyes from aqueous solution.

Science.gov (United States)

Takam, Brice; Acayanka, Elie; Kamgang, Georges Y; Pedekwang, Merlin T; Laminsi, Samuel

2017-07-01

Removal of cationic dye, Azur II, and anionic dye, Reactive Red 2 (RR-2) from aqueous solutions, has been successfully achieved by using a modified agricultural biomaterial waste: cocoa shell husk (Theobroma cacao) treated by gliding arc plasma (CPHP). The biomass in its natural form CPHN and modified form CPHP was characterized by infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), and point of zero charge (pH pzc ). Experimental variables such as initial pH, contact time, and temperature were optimized for adsorptive characteristics of CPHN and CPHP. The results show that the removal of the Azur II dye was favorable in the basic pH region (pH 10) while the Reactive Red 2 dye was favorable in the acidic pH region (pH 2). The minimum equilibrium time for Azur II and RR-2 dye was obtained after 40 and 240 min, respectively. The adsorption kinetics and isotherm data obtained were best described by a pseudo-second-order kinetic rate model and a combination of Langmuir-Freundlich isotherm models. This work indicates that the plasma-treated raw materials are good alternative multi-purpose sorbents for the removal of many coexisting pollutants from aqueous solutions.

A specific endogenous reference for genetically modified common bean (Phaseolus vulgaris L.) DNA quantification by real-time PCR targeting lectin gene.

Science.gov (United States)

Venturelli, Gustavo L; Brod, Fábio C A; Rossi, Gabriela B; Zimmermann, Naíra F; Oliveira, Jaison P; Faria, Josias C; Arisi, Ana C M

2014-11-01

The Embrapa 5.1 genetically modified (GM) common bean was approved for commercialization in Brazil. Methods for the quantification of this new genetically modified organism (GMO) are necessary. The development of a suitable endogenous reference is essential for GMO quantification by real-time PCR. Based on this, a new taxon-specific endogenous reference quantification assay was developed for Phaseolus vulgaris L. Three genes encoding common bean proteins (phaseolin, arcelin, and lectin) were selected as candidates for endogenous reference. Primers targeting these candidate genes were designed and the detection was evaluated using the SYBR Green chemistry. The assay targeting lectin gene showed higher specificity than the remaining assays, and a hydrolysis probe was then designed. This assay showed high specificity for 50 common bean samples from two gene pools, Andean and Mesoamerican. For GM common bean varieties, the results were similar to those obtained for non-GM isogenic varieties with PCR efficiency values ranging from 92 to 101 %. Moreover, this assay presented a limit of detection of ten haploid genome copies. The primers and probe developed in this work are suitable to detect and quantify either GM or non-GM common bean.

A modified RIA for minute albumin in human urine

International Nuclear Information System (INIS)

Chen Panzao; Hao Xiuhua; Xiao Shuqing; Li Zhenjia

1989-01-01

A modified radioimmunoassay for minute albuminuria using a solid phase radioiodination technique (Iodogen), and a precipitating reagent (PR) separation was described. The results of RIA and EIA of albumin are compared with each other (r = 0.925). Aliquots of 100μl diluted urine (1:20-1:100) are incubated at 4 deg C overnight with 100μl 125 I-labelled albumin and 100μl antiserum. Separation with 500 μl PR is very successful. The concentration of standard albumin ranges from 50 to 3200 ng/ml. The sensitivity of detection is 5 ng of albumin. The coefficients of inter-assay and intr-assay variation are 3.2-8.2% and 13.0-14.5% respectively. In 70 normal individuals the range of urinary albumin is 1.2-17.8 mg/24h

One Novel Multiple-Target Plasmid Reference Molecule Targeting Eight Genetically Modified Canola Events for Genetically Modified Canola Detection.

Science.gov (United States)

Li, Zhuqing; Li, Xiang; Wang, Canhua; Song, Guiwen; Pi, Liqun; Zheng, Lan; Zhang, Dabing; Yang, Litao

2017-09-27

Multiple-target plasmid DNA reference materials have been generated and utilized as good substitutes of matrix-based reference materials in the analysis of genetically modified organisms (GMOs). Herein, we report the construction of one multiple-target plasmid reference molecule, pCAN, which harbors eight GM canola event-specific sequences (RF1, RF2, MS1, MS8, Topas 19/2, Oxy235, RT73, and T45) and a partial sequence of the canola endogenous reference gene PEP. The applicability of this plasmid reference material in qualitative and quantitative PCR assays of the eight GM canola events was evaluated, including the analysis of specificity, limit of detection (LOD), limit of quantification (LOQ), and performance of pCAN in the analysis of various canola samples, etc. The LODs are 15 copies for RF2, MS1, and RT73 assays using pCAN as the calibrator and 10 genome copies for the other events. The LOQ in each event-specific real-time PCR assay is 20 copies. In quantitative real-time PCR analysis, the PCR efficiencies of all event-specific and PEP assays are between 91% and 97%, and the squared regression coefficients (R 2 ) are all higher than 0.99. The quantification bias values varied from 0.47% to 20.68% with relative standard deviation (RSD) from 1.06% to 24.61% in the quantification of simulated samples. Furthermore, 10 practical canola samples sampled from imported shipments in the port of Shanghai, China, were analyzed employing pCAN as the calibrator, and the results were comparable with those assays using commercial certified materials as the calibrator. Concluding from these results, we believe that this newly developed pCAN plasmid is one good candidate for being a plasmid DNA reference material in the detection and quantification of the eight GM canola events in routine analysis.

Selection of non-destructive assay methods: Neutron counting or calorimetric assay?

International Nuclear Information System (INIS)

Cremers, T.L.; Wachter, J.R.

1994-01-01

The transition of DOE facilities from production to D ampersand D has lead to more measurements of product, waste, scrap, and other less attractive materials. Some of these materials are difficult to analyze by either neutron counting or calorimetric assay. To determine the most efficacious analysis method, variety of materials, impure salts and hydrofluorination residues have been assayed by both calorimetric assay and neutron counting. New data will be presented together with a review of published data. The precision and accuracy of these measurements are compared to chemistry values and are reported. The contribution of the gamma ray isotopic determination measurement to the overall error of the calorimetric assay or neutron assay is examined and discussed. Other factors affecting selection of the most appropriate non-destructive assay method are listed and considered

Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability.

Science.gov (United States)

He, Dong; Luo, Wen; Wang, Zhiyuan; Lv, Pengmei; Yuan, Zhenhong; Huang, Shaowei; Xv, Jingliang

2017-07-01

Directed evolution has been proved an effective way to improve the stability of proteins, but high throughput screening assays for directed evolution with simultaneous improvement of two or more properties are still rare. In this study, we aimed to establish a membrane-blot assay for use in the high-throughput screening of Rhizomucor miehei lipases (RMLs). With the assistance of the membrane-blot screening assay, a mutant E47K named G10 that showed improved thermal stability was detected in the first round of error-prone PCR. Using G10 as the parent, two variants G10-11 and G10-20 that showed improved thermal stability and methanol tolerance without loss of activity compared to the wild type RML were obtained. The T 50 60 -value of G10-11 and G10-20 increased by 12°C and 6.5°C, respectively. After incubation for 1h, the remaining residual activity of G10-11 and G10-20 was 63.45% and 74.33%, respectively, in 50% methanol, and 15.98% and 30.22%, respectively, in 80% methanol. Thus, we successfully developed a membrane-blot assay that could be used for the high-throughput screening of RMLs with improved thermostability and methanol tolerance. Based on our findings, we believe that our newly developed membrane-blot assay will have potential applications in directed evolution in the future. Copyright © 2017 Elsevier Inc. All rights reserved.

A comparison of two colorimetric assays, based upon Lowry and Bradford techniques, to estimate total protein in soil extracts.

Science.gov (United States)

Redmile-Gordon, M A; Armenise, E; White, R P; Hirsch, P R; Goulding, K W T

2013-12-01

Soil extracts usually contain large quantities of dissolved humified organic material, typically reflected by high polyphenolic content. Since polyphenols seriously confound quantification of extracted protein, minimising this interference is important to ensure measurements are representative. Although the Bradford colorimetric assay is used routinely in soil science for rapid quantification protein in soil-extracts, it has several limitations. We therefore investigated an alternative colorimetric technique based on the Lowry assay (frequently used to measure protein and humic substances as distinct pools in microbial biofilms). The accuracies of both the Bradford assay and a modified Lowry microplate method were compared in factorial combination. Protein was quantified in soil-extracts (extracted with citrate), including standard additions of model protein (BSA) and polyphenol (Sigma H1675-2). Using the Lowry microplate assay described, no interfering effects of citrate were detected even with concentrations up to 5 times greater than are typically used to extract soil protein. Moreover, the Bradford assay was found to be highly susceptible to two simultaneous and confounding artefacts: 1) the colour development due to added protein was greatly inhibited by polyphenol concentration, and 2) substantial colour development was caused directly by the polyphenol addition. In contrast, the Lowry method enabled distinction between colour development from protein and non-protein origin, providing a more accurate quantitative analysis. These results suggest that the modified-Lowry method is a more suitable measure of extract protein (defined by standard equivalents) because it is less confounded by the high polyphenolic content which is so typical of soil extracts.

Assessment of angiogenic properties of biomaterials using the chicken embryo chorioallantoic membrane assay

International Nuclear Information System (INIS)

Azzarello, Joseph; Ihnat, Michael A; Kropp, Bradley P; Warnke, Linda A; Lin, H.-K.

2007-01-01

The angiogenic potential of a biomaterial is a critical factor for successful graft intake in tissue engineering. We developed a modified, rapid and reproducible chicken embryo chorioallantoic membrane (CAM) assay to evaluate the ability of biomaterials in inducing blood vessel density. Five biomaterials including one-layer porcine small intestinal submucosa (SIS), two-layer SIS, four-layer vacuum pressed (VP) SIS, polyglycolic acid (PGA) and PGA modified with poly(lactic-co-glycolic acid) (PLGA) were analyzed. A circular section (1.2 mm diameter) of each biomaterial was placed near a group of blood vessels in the CAM. Blood vessels around the biomaterials were captured with black and white images at 96 h post implantation; and the images were subjected to densitometry evaluation. One-layer SIS induced a significant increase in blood vessel density as compared to the cellulose nitrate negative control, and had the greatest increase in blood vessel density as compared to four-layer VP SIS, PGA, or PLGA modified PGA. Although two-layer SIS has enhanced physical structure for surgical manipulation, its induction in blood vessel density was significantly lower than the one-layer SIS. Stripping the SIS proteins or incubating one-layer SIS with neutralizing antibodies against basic fibroblast growth factor (bFGF) or vascular endothelial growth factor (VEGF) resulted in decreased angiogenesis. Consistent with results obtained from bladder augmentation animal models, these results confirmed that angiogenic growth factors were present in SIS and affected the angiogenic potential of biomaterials. These data also demonstrated that the CAM assay can be used to ascertain methodically the angiogenic potential of biomaterials

PCR-free quantitative detection of genetically modified organism from raw materials – A novel electrochemiluminescence-based bio-barcode method

Science.gov (United States)

Zhu, Debin; Tang, Yabing; Xing, Da; Chen, Wei R.

2018-01-01

Bio-barcode assay based on oligonucleotide-modified gold nanoparticles (Au-NPs) provides a PCR-free method for quantitative detection of nucleic acid targets. However, the current bio-barcode assay requires lengthy experimental procedures including the preparation and release of barcode DNA probes from the target-nanoparticle complex, and immobilization and hybridization of the probes for quantification. Herein, we report a novel PCR-free electrochemiluminescence (ECL)-based bio-barcode assay for the quantitative detection of genetically modified organism (GMO) from raw materials. It consists of tris-(2’2’-bipyridyl) ruthenium (TBR)-labele barcode DNA, nucleic acid hybridization using Au-NPs and biotin-labeled probes, and selective capture of the hybridization complex by streptavidin-coated paramagnetic beads. The detection of target DNA is realized by direct measurement of ECL emission of TBR. It can quantitatively detect target nucleic acids with high speed and sensitivity. This method can be used to quantitatively detect GMO fragments from real GMO products. PMID:18386909

Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

Directory of Open Access Journals (Sweden)

Carina Ladeira

2015-06-01

The results concerning of positive findings by micronuclei and non significant ones by comet assay, are corroborated by Deng et al. (2005 study performed in workers occupationally exposed to methotrexate, also a cytostatic drug. According to Cavallo et al. (2009, the comet assay seems to be more suitable for the prompt evaluation of the genotoxic effects, for instance, of polycyclic aromatic hydrocarbons mixtures containing volatile substances, whereas the micronucleus test seems more appropriate to evaluate the effects of exposure to antineoplastic agents. However, there are studies that observed an increase in both the comet assay and the micronucleus test in nurses handling antineoplastic drugs, although statistical significance was only seen in the comet assay, quite the opposite of our results (Maluf & Erdtmann, 2000; Laffon et al. 2005.

A modified fluorimetric host cell reactivation assay to determine the repair capacity of primary keratinocytes, melanocytes and fibroblasts

Directory of Open Access Journals (Sweden)

Gebhard Daniel

2010-06-01

Full Text Available Abstract Background The Host Cell Reactivation Assay (HCRA is widely used to identify circumstances and substances affecting the repair capacity of cells, however, it is restricted by the transfection procedure used and the sensitivity of the detection method. Primary skin cells are particularly difficult to transfect, and therefore sensitive methods are needed to detect any variations due to the cell-type or inter-individual differences or changes induced by diverse substances. A sensitive and repeatable method to detect the repair capacity of skin cells would be useful in two different aspects: On the one hand, to identify substances influencing the repair capacity in a positive manner (these substances could be promising ingredients for cosmetic products and on the other hand, to exclude the negative effects of substances on the repair capacity (this could serve as one step further towards replacing or at least reducing animal testing. Results In this paper, we present a rapid and sensitive assay to determine the repair capacity of primary keratinocytes, melanocytes and fibroblasts based on two wave-length Green Fluorescent Protein (GFP and DsRed reporter technology in order to test different substances and their potential to influence the DNA repair capacity. For the detection of plasmid restoration, we used FACS technology, which, in comparison to luminometer technology, is highly sensitive and allows single cell based analysis. The usefulness of this assay and studying the repair capacity is demonstrated by the evidence that DNA repair is repressed by Cyclosporin A in fibroblasts. Conclusions The methodology described in this paper determines the DNA repair capacity in different types of human skin cells. The described transfection protocol is suitable for the transfection of melanocytes, keratinocytes and fibroblasts, reaching efficacies suitable for the detection of the restored plasmids by FACS technology. Therefore the repair capacity

A multiplex bead-based suspension array assay for interrogation of phylogenetically informative single nucleotide polymorphisms for Bacillus anthracis

DEFF Research Database (Denmark)

Thierry, Simon; Hamidjaja, Raditijo A.; Girault, Guillaume

2013-01-01

been modified and adapted for simultaneous interrogation of 13 biallelic canonical SNPs in a 13-plex assay. Changes made to the originally published method include the design of allele-specific dual-priming-oligonucleotides (DPOs) as competing detection probes (MOLigo probes) and use of asymmetric PCR...

Development of a qualitative, multiplex real-time PCR kit for screening of genetically modified organisms (GMOs).

Science.gov (United States)

Dörries, Hans-Henno; Remus, Ivonne; Grönewald, Astrid; Grönewald, Cordt; Berghof-Jäger, Kornelia

2010-03-01

The number of commercially available genetically modified organisms (GMOs) and therefore the diversity of possible target sequences for molecular detection techniques are constantly increasing. As a result, GMO laboratories and the food production industry currently are forced to apply many different methods to reliably test raw material and complex processed food products. Screening methods have become more and more relevant to minimize the analytical effort and to make a preselection for further analysis (e.g., specific identification or quantification of the GMO). A multiplex real-time PCR kit was developed to detect the 35S promoter of the cauliflower mosaic virus, the terminator of the nopaline synthase gene of Agrobacterium tumefaciens, the 35S promoter from the figwort mosaic virus, and the bar gene of the soil bacterium Streptomyces hygroscopicus as the most widely used sequences in GMOs. The kit contains a second assay for the detection of plant-derived DNA to control the quality of the often processed and refined sample material. Additionally, the plant-specific assay comprises a homologous internal amplification control for inhibition control. The determined limits of detection for the five assays were 10 target copies/reaction. No amplification products were observed with DNAs of 26 bacterial species, 25 yeasts, 13 molds, and 41 not genetically modified plants. The specificity of the assays was further demonstrated to be 100% by the specific amplification of DNA derived from reference material from 22 genetically modified crops. The applicability of the kit in routine laboratory use was verified by testing of 50 spiked and unspiked food products. The herein described kit represents a simple and sensitive GMO screening method for the reliable detection of multiple GMO-specific target sequences in a multiplex real-time PCR reaction.

BioAssay templates for the semantic web

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Alex M. Clark

2016-05-01

Full Text Available Annotation of bioassay protocols using semantic web vocabulary is a way to make experiment descriptions machine-readable. Protocols are communicated using concise scientific English, which precludes most kinds of analysis by software algorithms. Given the availability of a sufficiently expressive ontology, some or all of the pertinent information can be captured by asserting a series of facts, expressed as semantic web triples (subject, predicate, object. With appropriate annotation, assays can be searched, clustered, tagged and evaluated in a multitude of ways, analogous to other segments of drug discovery informatics. The BioAssay Ontology (BAO has been previously designed for this express purpose, and provides a layered hierarchy of meaningful terms which can be linked to. Currently the biggest challenge is the issue of content creation: scientists cannot be expected to use the BAO effectively without having access to software tools that make it straightforward to use the vocabulary in a canonical way. We have sought to remove this barrier by: (1 defining a BioAssay Template (BAT data model; (2 creating a software tool for experts to create or modify templates to suit their needs; and (3 designing a common assay template (CAT to leverage the most value from the BAO terms. The CAT was carefully assembled by biologists in order to find a balance between the maximum amount of information captured vs. low degrees of freedom in order to keep the user experience as simple as possible. The data format that we use for describing templates and corresponding annotations is the native format of the semantic web (RDF triples, and we demonstrate some of the ways that generated content can be meaningfully queried using the SPARQL language. We have made all of these materials available as open source (http://github.com/cdd/bioassay-template, in order to encourage community input and use within diverse projects, including but not limited to our own

Enzyme activity assay of glycoprotein enzymes based on a boronate affinity molecularly imprinted 96-well microplate.

Science.gov (United States)

Bi, Xiaodong; Liu, Zhen

2014-12-16

Enzyme activity assay is an important method in clinical diagnostics. However, conventional enzyme activity assay suffers from apparent interference from the sample matrix. Herein, we present a new format of enzyme activity assay that can effectively eliminate the effects of the sample matrix. The key is a 96-well microplate modified with molecularly imprinted polymer (MIP) prepared according to a newly proposed method called boronate affinity-based oriented surface imprinting. Alkaline phosphatase (ALP), a glycoprotein enzyme that has been routinely used as an indicator for several diseases in clinical tests, was taken as a representative target enzyme. The prepared MIP exhibited strong affinity toward the template enzyme (with a dissociation constant of 10(-10) M) as well as superb tolerance for interference. Thus, the enzyme molecules in a complicated sample matrix could be specifically captured and cleaned up for enzyme activity assay, which eliminated the interference from the sample matrix. On the other hand, because the boronate affinity MIP could well retain the enzymatic activity of glycoprotein enzymes, the enzyme captured by the MIP was directly used for activity assay. Thus, additional assay time and possible enzyme or activity loss due to an enzyme release step required by other methods were avoided. Assay of ALP in human serum was successfully demonstrated, suggesting a promising prospect of the proposed method in real-world applications.

A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.

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Asif J Iqbal

Full Text Available Chemotaxis assays are an invaluable tool for studying the biological activity of inflammatory mediators such as CC chemokines, which have been implicated in a wide range of chronic inflammatory diseases. Conventional chemotaxis systems such as the modified Boyden chamber are limited in terms of the data captured given that the assays are analysed at a single time-point. We report the optimisation and validation of a label-free, real-time cell migration assay based on electrical cell impedance to measure chemotaxis of different primary murine macrophage populations in response to a range of CC chemokines and other chemoattractant signalling molecules. We clearly demonstrate key differences in the migratory behavior of different murine macrophage populations and show that this dynamic system measures true macrophage chemotaxis rather than chemokinesis or fugetaxis. We highlight an absolute requirement for Gαi signaling and actin cytoskeletal rearrangement as demonstrated by Pertussis toxin and cytochalasin D inhibition. We also studied the chemotaxis of CD14(+ human monocytes and demonstrate distinct chemotactic profiles amongst different monocyte donors to CCL2. This real-time chemotaxis assay will allow a detailed analysis of factors that regulate macrophage responses to chemoattractant cytokines and inflammatory mediators.

A Real Time Chemotaxis Assay Unveils Unique Migratory Profiles amongst Different Primary Murine Macrophages

Science.gov (United States)

Iqbal, Asif J.; Regan-Komito, Daniel; Christou, Ivy; White, Gemma E.; McNeill, Eileen; Kenyon, Amy; Taylor, Lewis; Kapellos, Theodore S.; Fisher, Edward A.; Channon, Keith M.; Greaves, David R.

2013-01-01

Chemotaxis assays are an invaluable tool for studying the biological activity of inflammatory mediators such as CC chemokines, which have been implicated in a wide range of chronic inflammatory diseases. Conventional chemotaxis systems such as the modified Boyden chamber are limited in terms of the data captured given that the assays are analysed at a single time-point. We report the optimisation and validation of a label-free, real-time cell migration assay based on electrical cell impedance to measure chemotaxis of different primary murine macrophage populations in response to a range of CC chemokines and other chemoattractant signalling molecules. We clearly demonstrate key differences in the migratory behavior of different murine macrophage populations and show that this dynamic system measures true macrophage chemotaxis rather than chemokinesis or fugetaxis. We highlight an absolute requirement for Gαi signaling and actin cytoskeletal rearrangement as demonstrated by Pertussis toxin and cytochalasin D inhibition. We also studied the chemotaxis of CD14+ human monocytes and demonstrate distinct chemotactic profiles amongst different monocyte donors to CCL2. This real-time chemotaxis assay will allow a detailed analysis of factors that regulate macrophage responses to chemoattractant cytokines and inflammatory mediators. PMID:23516549

Microbead agglutination based assays

KAUST Repository

Kodzius, Rimantas

2013-01-21

We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

An Improved Neutral a-Glucosidase Assay for Assessment of Epididymal Function—Validation and Comparison to the WHO Method

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Frank Eertmans

2014-01-01

Full Text Available Neutral a-glucosidase (NAG activity in human seminal plasma is an important indicator for epididymis functionality. In the present study, the classic World Health Organization (WHO method has been adapted to enhance assay robustness. Changes include modified enzyme reaction buffer composition and usage of an alternative enzyme inhibitor for background correction (glucose instead of castanospermine. Both methods have been tested in parallel on 144 semen samples, obtained from 94 patients/donors and 50 vasectomized men (negative control, respectively. Passing-Bablok regression analysis demonstrated equal assay performance. In terms of assay validation, analytical specificity, detection limit, measuring range, precision, and cut-off values have been calculated. These data confirm that the adapted method is a reliable, improved tool for NAG analysis in human semen.

Improved laboratory assays of Pu and U for SRP purification and finishing processes

International Nuclear Information System (INIS)

Holland, M.K.; Dorsett, R.S.

1986-01-01

Significant improvements have been made in routine assay techniques for uranium and plutonium as part of an effort to improve accountability at the Savannah River Plant (SRP). Emphasis was placed on input/output accountability points and key physical inventory tanks associated with purification and finishing processes. Improvements were made in existing assay methods; new methods were implemented; and the application of these methods was greatly expanded. Prior to assays, samples were validated via density measurements. Digital density meters precise to four, five, and six decimal places were used to meet specific requirements. Improved plutonium assay techniques are now in routine use: controlled-potential coulometry, ion-exchange coulometry, and Pu(III) diode-array spectrophotometry. A new state-of-the-art coulometer was fabricated and used to ensure maximum accuracy in verifying standards and in measuring plutonium in product streams. The diode-array spectrophotometer for Pu(III) measurements was modified with fiber optics to facilitate remote measurements; rapid, precise measurements made the technique ideally suited for high-throughput assays. For uranium assays, the isotope-dilution mass spectrometric (IDMS) method was converted to a gravimetric basis. The IDMS method and the existing Davies-Gray titration (gravimetric basis) have met accountability requirements for uranium. More recently, a Pu(VI) diode-array spectrophotometric method was used on a test basis to measure plutonium in shielded-cell input accountability samples. In addition, tests to measure uranium via diode-array spectrophotometry were initiated. This rapid, precise method will replace IDMS for certain key sample points

Advanced glycation end product (AGE) modified proteins in tears of diabetic patients.

Science.gov (United States)

Zhao, Zhenjun; Liu, Jingfang; Shi, Bingyin; He, Shuixiang; Yao, Xiaoli; Willcox, Mark D P

2010-08-11

High glucose level in diabetic patients may lead to advanced glycation end product (AGE) modified proteins. This study investigated AGE modified proteins in tears and compared their levels in diabetic patients (DM) with non-diabetic controls (CTL). Basal tears were collected from DM with (DR) or without (DNR) retinopathy and CTL. Total AGE modified proteins were detected quantitatively by a dot immunobinding assay. The AGE modified proteins were separated in 1D- and 2D-SDS gels and detected by western-blotting. The individual AGE modified proteins were also compared between groups using densitometry. Compared with the CTL group, tear concentrations of AGE modified proteins were significantly elevated in DR and DNR groups. The concentration of AGE modified proteins in diabetic tears were positively correlated with AGE modified hemoglobin (HbA1c) and postprandial blood glucose level (PBG). Western blotting of AGE modified proteins from 1D-SDS gels showed several bands, the major one at around 60 kDa. The intensities of AGE modified protein bands were higher in DM tears than in CTL tears. Western blotting from 2D-SDS gels showed a strongly stained horizontal strip, which corresponded to the major band in 1D-SDS gels. Most of the other AGE modified protein species were within molecular weight of 30-60 kDa, PI 5.2-7.0. Densitometry analysis demonstrated several AGE modified proteins were elevated in DR or DNR tears. Total and some individual AGE modified proteins were elevated in DM tears. AGE modified proteins in tears may be used as biomarkers to diagnose diabetes and/or diabetic retinopathy.

Development of laser-induced fluorescence detection to assay DNA damage

International Nuclear Information System (INIS)

Sharma, M.; Freund, H.G.

1991-01-01

A precolumn derivation method has been developed for high performance liquid chromatographic (HPLC) analysis of DNA damage using fluorescence detection. The modified nucleotide, having excised enzymatically from the exposed DNA, is enriched from the normal nucleotides and labeled with a fluorescent reagent. The labeling procedure involves phosphoramidation of the nucleotide with ethylenediamine (EDA) followed by conjugation of the free amino end of the phosphoramidate with 5-dimethylaminonaphthalene 1-sulfonyl chloride, commonly known as Dansyl chloride. The dansylated nucleotide can be analyzed with a sub-picomole limit of detection (LOD) by conventional HPLC using a conventional fluorescence detector. By combining microbore HPLC with laser-induced fluorescence (LIF) detection, the authors present the development of an analytical system that has sub-femtomole LOD for real-time analysis of the dansylated nucleotide. In this paper the application of the developed system in fluorescence postlabeling assay of a small alkyl-modified nucleotide (5-methyl CMP) in calf-thymus DNA is discussed

An SH2 domain-based tyrosine kinase assay using biotin ligase modified with a terbium(III) complex.

Science.gov (United States)

Sueda, Shinji; Shinboku, Yuki; Kusaba, Takeshi

2013-01-01

Src homology 2 (SH2) domains are modules of approximately 100 amino acids and are known to bind phosphotyrosine-containing sequences with high affinity and specificity. In the present work, we developed an SH2 domain-based assay for Src tyrosine kinase using a unique biotinylation reaction from archaeon Sulfolobus tokodaii. S. tokodaii biotinylation has a unique property that biotin protein ligase (BPL) forms a stable complex with its biotinylated substrate protein (BCCP). Here, an SH2 domain from lymphocyte-specific tyrosine kinase was genetically fused to a truncated BCCP, and the resulting fusion protein was labeled through biotinylation with BPL carrying multiple copies of a luminescent Tb(3+) complex. The labeled SH2 fusion proteins were employed to detect a phosphorylated peptide immobilized on the surface of the microtiter plate, where the phosphorylated peptide was produced by phosphorylation to the substrate peptide by Src tyrosine kinase. Our assay allows for a reliable determination of the activity of Src kinase lower than 10 pg/μL by a simple procedure.

Seismogenic rotational slumps and translational glides in pelagic deep-water carbonates. Upper Tithonian-Berriasian of Southern Tethyan margin (W Sicily, Italy)

Science.gov (United States)

Basilone, Luca

2017-07-01

Soft-sediment deformation structures (SSDSs), which reflect sediment mobilization processes, are helpful to identify punctual events of paleoenvironmental stresses. In the upper Tithonian-Berriasian calpionellid pelagic limestone of the Lattimusa Fm. outcropping in the Barracù section (W Sicily), paleoenvironmental restoration reveals the occurrence of a deep-water flat basin, characterised by undeformed planar bedding, laterally passing to a gentle slope where the deformed horizons alternate with undeformed beds. Here, two types of gravity slides have been differentiated on the basis of different kinds of SSDSs, brittle deformation, involved lithofacies, geometry and morphology. Type 1 slump sheets, involving marl-limestone couplets, are characterised by chaotic stratification, truncated beds, recumbent folds, tension gashes, clastic dikes and small-scale listric normal faults and thrusts. They moved downslope as a rotational slump, displaying the typical wedge-shaped morphology, becoming thicker toe-wards. Type 2 slump sheets, involving bed packages of lime mudstone, are characterised by arcuate slump scars developing downwards as listric normal faults, associated with rotational beds and bulging. Displaying tabular geometries, they moved coherently along an overall bedding parallel detachment surface as a translational glide characterised by sediment creep. Regional geology indicates that the basin (Sicanian), bordered by a stepped carbonate platform, was affected by synsedimentary tectonics throughout the Mesozoic. The synsedimentary extensional tectonics that affect the Upper Triassic-Jurassic deposits with steeped normal faults, causing tilt-block and instability of the seafloor through fluidization processes, triggered the rotational slumps of the lower portion of the section. Seismic shocks, induced by outside sector tectonics, like those recorded in the adjacent Busambra stepped carbonate platform margin, triggered the rotational slumps that are not

Neuromyelitis optica immunoglobulin G in Chinese patients detected by immunofluorescence assay on a monkey brain substrate.

Science.gov (United States)

Long, Youming; Hu, Xueqiang; Peng, Fuhua; Lu, Zhengqi; Wang, Yuge; Yang, Yu; Qiu, Wei

2012-01-01

Serum neuromyelitis optica immunoglobulin G (NMO-IgG) is used as a biomarker to differentiate between neuromyelitis optica (NMO) and multiple sclerosis (MS). However, the original assay is expensive and complex and shows low sensitivity. Here, we investigated the potential of NMO-IgG detection using an indirect immunofluorescence (IIF) assay on monkey brains. NMO-IgG seroprevalence was determined in 168 samples by an IIF assay on a monkey brain substrate. The data were compared with those from a standard mouse brain IIF assay using McNemar and kappa tests. Thirty-one of 50 (62%) NMO patients, 7 of 18 (38.9%) longitudinally extensive transverse myelitis patients, 6 of 57 (10.5%) MS patients, and 5 of 10 (50%) optic neuritis patients were seropositive for NMO-IgG. None of the acute partial transverse myelitis patients (n = 3) or healthy controls (n = 20) was positive. Thus, the sensitivity of the test was 62% for the patients with clinically definite NMO. The specificity was 89.5%, considering the 57 MS patients as the control group. The modified IIF assay on monkey brains and the standard IIF assay based on mouse brains were not significantly different (McNemar test; p = 1.000). The two assays were concordant in 39 seropositive samples and 100 seronegative samples (kappa test; kappa = 0.592, p monkey brain assay was no better than the standard mouse brain IIF assay, we affirmed that NMO-IgG is a sensitive and specific biomarker to differentiate between NMO and MS. Copyright © 2011 S. Karger AG, Basel.

Utility of loop-mediated isothermal amplification assay, polymerase chain reaction, and elisa for diagnosis of leptospirosis in South Indian patients

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Mallika Sengupta

2017-01-01

Full Text Available Background: Leptospirosis is a zoonotic disease which requires laboratory diagnosis for confirmation. Materials and Methods: In this study serum samples from adults with acute undifferentiated fever (duration ≤15 days were tested for IgM antibodies to Leptospira by ELISA, PCR for rrs gene and loop-mediated isothermal amplification (LAMP assay for LipL32 and LipL41. Results: Among the 150 sera tested, three were positive by PCR, LAMP and IgM ELISA/modified Faines' criteria, two by only PCR; seven only by LAMP assay and forty fulfilled modified Faine's criteria (illness clinically compatible and IgM ELISA positive for leptospirosis. Clinical correlation revealed renal compromise, low platelet count and severe jaundice were significantly related to leptospirosis (P < 0.05. Conclusion: This study suggests that LAMP assay could be useful for diagnosis of leptospirosis during the 1st week of illness whereas IgM ELISA forms the mainstay of diagnosis from the 2nd week onward. Further studies especially community based, comparing ELISA, PCR, LAMP, culture and microscopic agglutination test are required to evaluate the veracity of these findings.

Critical thickness of atomically ordered III-V alloys

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France, R. M.; McMahon, W. E.; Guthrey, H. L. [National Renewable Energy Laboratory, 15013 Denver West Parkway, Golden, Colorado 80401 (United States)

2015-10-12

The critical thickness model is modified with a general boundary energy that describes the change in bulk energy as a dislocation regularly alters the atomic structure of an ordered material. The model is evaluated for dislocations gliding through CuPt-ordered GaInP and GaInAs, where the boundary energy is negative and the boundary is stable. With ordering present, the critical thickness is significantly lowered and remains finite as the mismatch strain approaches zero. The reduction in critical thickness is most significant when the order parameter is greatest and the amount of misfit energy is low. The modified model is experimentally validated for low-misfit GaInP epilayers with varying order parameters using in situ wafer curvature and ex situ cathodoluminescence. With strong ordering, relaxation begins at a lower thickness and occurs at a greater rate, which is consistent with a lower critical thickness and increased glide force. Thus, atomic ordering is an important consideration for the stability of lattice-mismatched devices.

Performance evaluation of Cepheid Xpert Norovirus kit with a user-modified protocol.

Science.gov (United States)

Wong, Rachel Shi-Lei; Yeo, Fion; Chia, Wai Theng; Lee, Chun Kiat; Leong, Mun Han; Ng, Christopher Wai-Siong; Poon, Kok Siong; Yan, Gabriel Zherong; Chiu, Lily-Lily; Yan, Benedict Junrong; Jureen, Roland; Koay, Evelyn Siew-Chuan; Lee, Hong Kai

2018-03-01

The Cepheid Xpert® Norovirus kit automates sample processing, nucleic acid extraction, and real-time reverse transcription polymerase chain reactions (RT-PCRs) to detect norovirus genogroups I (GI) and II (GII). Eighty-five stool samples collected between February 2015 and May 2017 were used to compare the performance of a user-modified Xpert assay against a clinically validated laboratory-developed test (LDT). Of the 85 samples, 54 were previously archived in -80°C freezer. The remaining 31 were fresh samples tested concurrently with the LDT. The results of all samples tested using the Xpert kit and LDT were found to be concordant, including 12 GI- and 42 GII-positive samples, 1 GI and GII coinfection, and 30 negative samples. Comparison of the assays showed perfect concordance with a kappa coefficient score of 1.00 (95%CI from 1.00 to 1.00). Of the 30 negative stool samples tested, three samples were positive for rotavirus detected using an immunochromatographic assay, with no cross-reactivity shown in both LDT and Xpert assays. In-run sample processing control of the Xpert assay for all negative samples tested showed no/minor inhibition. Compared to the LDT, the Xpert assay produced similar or better Ct values for detection. It also showed better mitigation of PCR inhibition in stool sample testing. © 2017 Wiley Periodicals, Inc.

High-throughput kinase assays with protein substrates using fluorescent polymer superquenching

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Weatherford Wendy

2005-05-01

Full Text Available Abstract Background High-throughput screening is used by the pharmaceutical industry for identifying lead compounds that interact with targets of pharmacological interest. Because of the key role that aberrant regulation of protein phosphorylation plays in diseases such as cancer, diabetes and hypertension, kinases have become one of the main drug targets. With the exception of antibody-based assays, methods to screen for specific kinase activity are generally restricted to the use of small synthetic peptides as substrates. However, the use of natural protein substrates has the advantage that potential inhibitors can be detected that affect enzyme activity by binding to a site other than the catalytic site. We have previously reported a non-radioactive and non-antibody-based fluorescence quench assay for detection of phosphorylation or dephosphorylation using synthetic peptide substrates. The aim of this work is to develop an assay for detection of phosphorylation of chemically unmodified proteins based on this polymer superquenching platform. Results Using a modified QTL Lightspeed™ assay, phosphorylation of native protein was quantified by the interaction of the phosphorylated proteins with metal-ion coordinating groups co-located with fluorescent polymer deposited onto microspheres. The binding of phospho-protein inhibits a dye-labeled "tracer" peptide from associating to the phosphate-binding sites present on the fluorescent microspheres. The resulting inhibition of quench generates a "turn on" assay, in which the signal correlates with the phosphorylation of the substrate. The assay was tested on three different proteins: Myelin Basic Protein (MBP, Histone H1 and Phosphorylated heat- and acid-stable protein (PHAS-1. Phosphorylation of the proteins was detected by Protein Kinase Cα (PKCα and by the Interleukin -1 Receptor-associated Kinase 4 (IRAK4. Enzyme inhibition yielded IC50 values that were comparable to those obtained using

High-throughput kinase assays with protein substrates using fluorescent polymer superquenching.

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Rininsland, Frauke; Stankewicz, Casey; Weatherford, Wendy; McBranch, Duncan

2005-05-31

High-throughput screening is used by the pharmaceutical industry for identifying lead compounds that interact with targets of pharmacological interest. Because of the key role that aberrant regulation of protein phosphorylation plays in diseases such as cancer, diabetes and hypertension, kinases have become one of the main drug targets. With the exception of antibody-based assays, methods to screen for specific kinase activity are generally restricted to the use of small synthetic peptides as substrates. However, the use of natural protein substrates has the advantage that potential inhibitors can be detected that affect enzyme activity by binding to a site other than the catalytic site. We have previously reported a non-radioactive and non-antibody-based fluorescence quench assay for detection of phosphorylation or dephosphorylation using synthetic peptide substrates. The aim of this work is to develop an assay for detection of phosphorylation of chemically unmodified proteins based on this polymer superquenching platform. Using a modified QTL Lightspeed assay, phosphorylation of native protein was quantified by the interaction of the phosphorylated proteins with metal-ion coordinating groups co-located with fluorescent polymer deposited onto microspheres. The binding of phospho-protein inhibits a dye-labeled "tracer" peptide from associating to the phosphate-binding sites present on the fluorescent microspheres. The resulting inhibition of quench generates a "turn on" assay, in which the signal correlates with the phosphorylation of the substrate. The assay was tested on three different proteins: Myelin Basic Protein (MBP), Histone H1 and Phosphorylated heat- and acid-stable protein (PHAS-1). Phosphorylation of the proteins was detected by Protein Kinase Calpha (PKCalpha) and by the Interleukin -1 Receptor-associated Kinase 4 (IRAK4). Enzyme inhibition yielded IC50 values that were comparable to those obtained using peptide substrates. Statistical parameters that

Electrochemical sensor for multiplex screening of genetically modified DNA: identification of biotech crops by logic-based biomolecular analysis.

Science.gov (United States)

Liao, Wei-Ching; Chuang, Min-Chieh; Ho, Ja-An Annie

2013-12-15

Genetically modified (GM) technique, one of the modern biomolecular engineering technologies, has been deemed as profitable strategy to fight against global starvation. Yet rapid and reliable analytical method is deficient to evaluate the quality and potential risk of such resulting GM products. We herein present a biomolecular analytical system constructed with distinct biochemical activities to expedite the computational detection of genetically modified organisms (GMOs). The computational mechanism provides an alternative to the complex procedures commonly involved in the screening of GMOs. Given that the bioanalytical system is capable of processing promoter, coding and species genes, affirmative interpretations succeed to identify specified GM event in terms of both electrochemical and optical fashions. The biomolecular computational assay exhibits detection capability of genetically modified DNA below sub-nanomolar level and is found interference-free by abundant coexistence of non-GM DNA. This bioanalytical system, furthermore, sophisticates in array fashion operating multiplex screening against variable GM events. Such a biomolecular computational assay and biosensor holds great promise for rapid, cost-effective, and high-fidelity screening of GMO. Copyright © 2013 Elsevier B.V. All rights reserved.

Improving colorimetric assays through protein enzyme-assisted gold nanoparticle amplification.

Science.gov (United States)

Xie, Xiaoji; Xu, Wei; Liu, Xiaogang

2012-09-18

The discovery of the DNA-mediated assembly of gold nanoparticles was a great moment in the history of science; this understanding and chemical control enabled the rational design of functional nanomaterials as novel probes in biodetection. In contrast with conventional probes such as organic dyes, gold nanoparticles exhibit high photostability and unique size-dependent optical properties. Because of their high extinction coefficients and strong distance dependent optical properties, these nanoparticles have emerged over the past decade as a promising platform for rapid, highly sensitive colorimetric assays that allow for the visual detection of low concentrations of metal ions, small molecules, and biomacromolecules. These discoveries have deepened our knowledge of biological phenomena and facilitated the development of many new diagnostic and therapeutic tools. Despite these many advances and continued research efforts, current nanoparticle-based colorimetric detection systems still suffer from several drawbacks, such as limited sensitivity and selectivity. This Account describes the recent development of colorimetric assays based on protein enzyme-assisted gold nanoparticle amplification. The benefits of such detection systems include significantly improved detection sensitivity and selectivity. First, we discuss the general design of enzyme-modified nanoparticle systems in colorimetric assays. We show that a quantitative understanding of the unique properties of different enzymes is paramount for effective biological assays. We then examine the assays for nucleic acid detection based on different types of enzymes, including endonucleases, ligases, and polymerases. For each of these assays, we identify the underlying principles that contribute to the enhanced detection capability of nanoparticle systems and illustrate them with selected examples. Furthermore, we demonstrate that the combination of gold nanoparticles and specific enzymes can probe enzyme dynamics

Novel Route for Agmatine Catabolism in Aspergillus niger: 4-Guanidinobutyrase Assay.

Science.gov (United States)

Saragadam, Tejaswani; Punekar, Narayan S

2018-01-01

The enzyme 4-guanidinobutyrase (GBase) catalyzes the hydrolysis of 4-guanidinobutyric acid (GB) to 4-aminobutyric acid (GABA) and urea. Here we describe methods to estimate urea and GABA that were suitably adapted from the published literature. The urea is determined by colorimetric assay using modified Archibald's method. However, the low sensitivity of this method often renders it impractical to perform fine kinetic analysis. To overcome this limitation, a high sensitive method for detecting GABA is exploited that can even detect 1 μM of GABA in the assay mixture. The samples are deproteinized by perchloric acid (PCA) and potassium hydroxide treatment prior to HPLC analysis of GABA. The method involves a pre-column derivatization with o-phthalaldehyde (OPA) in combination with the thiol 3-mercaptopropionic acid (MPA). The fluorescent GABA derivative is then detected after reversed phase high performance liquid chromatography (RP-HPLC) using isocratic elution. The protocols described here are broadly applicable to other biological samples involving urea and GABA as metabolites.

Estimates of DNA damage by the comet assay in the direct-developing frog Eleutherodactylus johnstonei (Anura, Eleutherodactylidae

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Laura Carolina Valencia

2011-01-01

Full Text Available The aim of this study was to use the Comet assay to assess genetic damage in the direct-developing frog Eleutherodactylus johnstonei. A DNA diffusion assay was used to evaluate the effectiveness of alkaline, enzymatic and alkaline/enzymatic treatments for lysing E. johnstonei blood cells and to determine the amount of DNA strand breakage associated with apoptosis and necrosis. Cell sensitivity to the mutagens bleomycin (BLM and 4-nitroquinoline-1-oxide (4NQO was also assessed using the Comet assay, as was the assay reproducibility. Alkaline treatment did not lyse the cytoplasmic and nuclear membranes of E. johnstonei blood cells, whereas enzymatic digestion with proteinase K (40 !g/mL yielded naked nuclei. The contribution of apoptosis and necrosis (assessed by the DNA diffusion assay to DNA damage was estimated to range from 0% to 8%. BLM and 4NQO induced DNA damage in E. johnstonei blood cells at different concentrations and exposure times. Dose-effect curves with both mutagens were highly reproducible and showed consistently low coefficients of variation (CV < 10%. The results are discussed with regard to the potential use of the modified Comet assay for assessing the exposure of E. johnstonei to herbicides in ecotoxicological studies.

Estimates of DNA damage by the comet assay in the direct-developing frog Eleutherodactylus johnstonei (Anura, Eleutherodactylidae).

Science.gov (United States)

Valencia, Laura Carolina; García, Adriana; Ramírez-Pinilla, Martha Patricia; Fuentes, Jorge Luis

2011-10-01

The aim of this study was to use the Comet assay to assess genetic damage in the direct-developing frog Eleutherodactylus johnstonei. A DNA diffusion assay was used to evaluate the effectiveness of alkaline, enzymatic and alkaline/enzymatic treatments for lysing E. johnstonei blood cells and to determine the amount of DNA strand breakage associated with apoptosis and necrosis. Cell sensitivity to the mutagens bleomycin (BLM) and 4-nitro-quinoline-1-oxide (4NQO) was also assessed using the Comet assay, as was the assay reproducibility. Alkaline treatment did not lyse the cytoplasmic and nuclear membranes of E. johnstonei blood cells, whereas enzymatic digestion with proteinase K (40 μg/mL) yielded naked nuclei. The contribution of apoptosis and necrosis (assessed by the DNA diffusion assay) to DNA damage was estimated to range from 0% to 8%. BLM and 4NQO induced DNA damage in E. johnstonei blood cells at different concentrations and exposure times. Dose-effect curves with both mutagens were highly reproducible and showed consistently low coefficients of variation (CV ≤ 10%). The results are discussed with regard to the potential use of the modified Comet assay for assessing the exposure of E. johnstonei to herbicides in ecotoxicological studies.

Huntingtin interacting proteins are genetic modifiers of neurodegeneration.

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Linda S Kaltenbach

2007-05-01

Full Text Available Huntington's disease (HD is a fatal neurodegenerative condition caused by expansion of the polyglutamine tract in the huntingtin (Htt protein. Neuronal toxicity in HD is thought to be, at least in part, a consequence of protein interactions involving mutant Htt. We therefore hypothesized that genetic modifiers of HD neurodegeneration should be enriched among Htt protein interactors. To test this idea, we identified a comprehensive set of Htt interactors using two complementary approaches: high-throughput yeast two-hybrid screening and affinity pull down followed by mass spectrometry. This effort led to the identification of 234 high-confidence Htt-associated proteins, 104 of which were found with the yeast method and 130 with the pull downs. We then tested an arbitrary set of 60 genes encoding interacting proteins for their ability to behave as genetic modifiers of neurodegeneration in a Drosophila model of HD. This high-content validation assay showed that 27 of 60 orthologs tested were high-confidence genetic modifiers, as modification was observed with more than one allele. The 45% hit rate for genetic modifiers seen among the interactors is an order of magnitude higher than the 1%-4% typically observed in unbiased genetic screens. Genetic modifiers were similarly represented among proteins discovered using yeast two-hybrid and pull-down/mass spectrometry methods, supporting the notion that these complementary technologies are equally useful in identifying biologically relevant proteins. Interacting proteins confirmed as modifiers of the neurodegeneration phenotype represent a diverse array of biological functions, including synaptic transmission, cytoskeletal organization, signal transduction, and transcription. Among the modifiers were 17 loss-of-function suppressors of neurodegeneration, which can be considered potential targets for therapeutic intervention. Finally, we show that seven interacting proteins from among 11 tested were able to

Assay-specific decision limits for two new automated parathyroid hormone and 25-hydroxyvitamin D assays.

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Souberbielle, Jean-Claude; Fayol, Véronique; Sault, Corinne; Lawson-Body, Ethel; Kahan, André; Cormier, Catherine

2005-02-01

The recent development of nonradioactive automated assays for serum parathyroid hormone (PTH) and 25-hydroxyvitamin D (25OHD) has made measurement of these two hormones possible in many laboratories. In this study, we compared two new assays for PTH and 25OHD adapted on an automated analyzer, the LIAISON, with two manual immunoassays used worldwide. We studied 228 osteoporotic patients, 927 healthy individuals, 38 patients with primary hyperparathyroidism, and 167 hemodialyzed patients. Serum PTH was measured with the Allegro and the LIAISON assays, and 25OHD was measured with DiaSorin RIA and the LIAISON assay. Regression analysis was used to calculate decision thresholds for the LIAISON assays that were equivalent to those of the Allegro PTH and DiaSorin 25OHD assays. The 25OHD concentrations obtained with the LIAISON assay and the RIA in osteoporotic patients were well correlated (r = 0.83; P 50 nmol/L as eligible for the reference population for the LIAISON PTH assay. In this group, the 3rd-97th percentile interval for LIAISON PTH was 3-51 ng/L. Considering upper reference limits of 46 and 51 ng/L for the Allegro and LIAISON assays, respectively, the frequency of above-normal PTH concentrations in patients with primary hyperparathyroidism was similar in both assays. Regression analysis between serum PTH measured by the Allegro and LIAISON assays in 167 hemodialyzed patients and the corresponding Bland-Altman analysis of these data suggest that the LIAISON PTH assay tends to read higher than the Allegro assay at low concentrations but lower at high concentrations (>300 ng/L). Because clinical decision limits for both PTH and 25OHD should be assay specific, we propose equivalences between these assays and two manual assays used worldwide. These assay-specific decision limits should help potential users of the LIAISON PTH and 25OHD assays.

Comparison of Two Assays to Determine Anti-Citrullinated Peptide Antibodies in Rheumatoid Arthritis in relation to Other Chronic Inflammatory Rheumatic Diseases: Assaying Anti-Modified Citrullinated Vimentin Antibodies Adds Value to Second-Generation Anti-Citrullinated Cyclic Peptides Testing

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Miriam Lizette Díaz-Toscano

2014-01-01

Full Text Available Determination of anti-citrullinated peptide antibodies (ACPA plays a relevant role in the diagnosis of rheumatoid arthritis (RA. To date, it is still unclear if the use of several tests for these autoantibodies in the same patient offers additional value as compared to performing only one test. Therefore, we evaluated the performance of using two assays for ACPA: second-generation anti-citrullinated cyclic peptides antibodies (anti-CCP2 and anti-mutated citrullinated vimentin (anti-MCV antibodies for the diagnosis of RA. We compared three groups: RA (n=142, chronic inflammatory disease (CIRD, n=86, and clinically healthy subjects (CHS, n=56 to evaluate sensitivity, specificity, predictive values, and likelihood ratios (LR of these two assays for the presence of RA. A lower frequency of positivity for anti-CCP2 was found in RA (66.2% as compared with anti-MCV (81.0%. When comparing RA versus other CIRD, sensitivity increased when both assays were performed. This strategy of testing both assays had high specificity and LR+. We conclude that adding the assay of anti-MCV antibodies to the determination of anti-CCP2 increases the sensitivity for detecting seropositive RA. Therefore, we propose the use of both assays in the initial screening of RA in longitudinal studies, including early onset of undifferentiated arthritis.

Adaptation of the neutral bacterial comet assay to assess antimicrobial-mediated DNA double-strand breaks in Escherichia coli

Science.gov (United States)

SOLANKY, DIPESH; HAYDEL, SHELLEY E.

2012-01-01

This study aimed to determine the mechanism of action of a natural antibacterial clay mineral mixture, designated CB, by investigating the induction of DNA double-strand breaks (DSBs) in Escherichia coli. To quantify DNA damage upon exposure to soluble antimicrobial compounds, we modified a bacterial neutral comet assay, which primarily associates the general length of an electrophoresed chromosome, or comet, with the degree of DSB-associated DNA damage. To appropriately account for antimicrobial-mediated strand fragmentation, suitable control reactions consisting of exposures to water, ethanol, kanamycin, and bleomycin were developed and optimized for the assay. Bacterial exposure to the CB clay resulted in significantly longer comet lengths, compared to water and kanamycin exposures, suggesting that the induction of DNA DSBs contributes to the killing activity of this antibacterial clay mineral mixture. The comet assay protocol described herein provides a general technique for evaluating soluble antimicrobial-derived DNA damage and for comparing DNA fragmentation between experimental and control assays. PMID:22940101

Correspondence between radioactive and functional methods in the quality control of DNA restriction and modifying enzymes

DEFF Research Database (Denmark)

Trujillo, L E; Pupo, E; Miranda, F

1996-01-01

We evaluated the use of two radiolabeled lambda DNA/Hpa II substrates to detect 5'-->3', 3'-->5' single and double stranded DNA dependent exonuclease and phosphatase activities found as contaminants in restriction and modifying enzyme preparations. Looking for the meaning of the radioactive assay...

Evaluation of assays for quantification of DNA in canine plasma as an indirect marker of NETosis.

Science.gov (United States)

Smith, Stephanie A; Lawson, Corinne M; McMichael, Maureen A; Jung, Katrina; O'Brien, Mauria; Achiel, Ron

2017-06-01

Neutrophil extracellular traps (NET), consisting of a filamentous DNA/chromatin-histone scaffold originating from neutrophils are part of the innate immune response, may be released under a variety of inflammatory conditions and are associated with an increased risk for thrombosis. The purpose of this study was to evaluate a SYTOX green fluorescence assay and a histone-DNA complex (hisDNA) ELISA for quantification of NET-related DNA in canine plasma. The influence of variations in blood sample handling on assay results was tested. Accuracy of the SYTOX green fluorescence and the hisDNA ELISA was evaluated with dilutional linearity using serial dilutions. Interference was assessed by addition of purified bilirubin or hemoglobin. Precision was determined by calculating the intra- and inter-assay CV. Preanalytic sample handling did not influence DNA measurements by either assay. Citrate and EDTA plasma samples were equivalent. For the DNA fluorescence assay, dilutional linearity was poor due to autofluorescence, which was corrected by addition of canine plasma to the diluent. The presence of bilirubin and hemoglobin also increased autofluorescence, and resulted in falsely low concentrations of DNA. On the hisDNA ELISA, pigmentemia had no effect. Both assays as modified in this study are suitable for measuring DNA in canine EDTA or citrate plasma. However, performance of the fluorescence assay was impacted by pigmentemia, and it was less sensitive than the ELISA in detecting the presence of nucleosome material in the plasma. © 2017 American Society for Veterinary Clinical Pathology.

Cell-mediated cytotoxicity for melanome tumor cells: detection by a (3H)proline release assay

International Nuclear Information System (INIS)

Saal, J.G.; Rieber, E.P.; Riethmueller, G.

1976-01-01

An in vitro lymphocyte-mediated cytotoxicity assay using [ 3 H]proline-labelled target cells is described. The assay, modified from an original procedure of Bean et al., assesses the release of [ 3 H]proline by filtering the total culture fluid containing both trypsinised tumor cells and effector cells. Filtration is performed with a semiautomatic harvesting device using low suction pressure and large-diameter glass filters. Pretreatment of filters with whole serum diminishes adsorption of cell-free radioactive material considerably and thus increases the sensitivity of the assay. Nearly 100% of the radioactivity could be recovered with this harvesting device. The technique allowed the detection of cytolytic activities of lymphocytes after 6 h of incubation. Lymphocytes from patients with primary malignant melanoma showed a significantly higher cytolytic reactivity (p > 0.001) than normal donors' lymphocytes against three different melanoma cell lines. In a series of parallel experiments on 36 patients and 18 normal donors, this modification of the [ 3 ]proline test was compared with three different assays: the conventional microcytotoxicity test of Takasugi and Klein, the original [ 3 H]proline microcytotoxicity test of Bean et al., and the viability count of tumor cells. (Auth.)

LNA-modified oligonucleotides mediate specific inhibition of microRNA function

DEFF Research Database (Denmark)

Ørom, Ulf Andersson; Kauppinen, Sakari; Lund, Anders H

2006-01-01

microRNAs are short, endogenous non-coding RNAs that act as post-transcriptional modulators of gene expression. Important functions for microRNAs have been found in the regulation of development, cellular proliferation and differentiation, while perturbed miRNA expression patterns have been...... observed in many human cancers. Here we present a method for specific inhibition of miRNA function through interaction with LNA-modified antisense oligonucleotides and report the specificity of this application. We show that LNA-modified oligonucleotides can inhibit exogenously introduced miRNAs with high...... specificity using a heterologous reporter assay, and furthermore demonstrate their ability to inhibit an endogenous miRNA in Drosophila melanogaster cells, leading to up-regulation of the cognate target protein. The method shows stoichiometric and reliable inhibition of the targeted miRNA and can thus...

Biomonitoring of genotoxic risk in radar facility workers: comparison of the comet assay with micronucleus assay and chromatid breakage assay

International Nuclear Information System (INIS)

Garaj-Vrhovac, V.; Kopjar, N.

2003-01-01

Genotoxic risks of occupational exposure in a radar facility were evaluated by using alkaline comet assay, micronucleus assay and chromatid breakage assay on peripheral blood leukocytes in exposed subjects and corresponding controls. Results show that occupational exposure to microwave radiation correlates with an increase of genome damage in somatic cells. The levels of DNA damage in exposed subjects determined by using alkaline comet assay were increased compared to control and showed interindividual variations. Incidence of micronuclei was also significantly increased compared to baseline control values. After short exposure of cultured lymphocytes to bleomycin, cells of occupationally exposed subjects responded with high numbers of chromatid breaks. Although the level of chromosome damage generated by bleomycin varied greatly between individuals, in exposed subjects a significantly elevated number of chromatid breaks was observed. Our results support data reported in literature indicating that microwave radiation represents a potential DNA-damaging hazard. Alkaline comet assay is confirmed as a sensitive and highly reproducible technique for detection of primary DNA damage inflicted in somatic cells. Micronucleus assay was confirmed as reliable bio-markers of effect and chromatid breakage assay as sensitive bio-marker of individual cancer susceptibility. The results obtained also confirm the necessity to improve measures and to perform accurate health surveillance of individuals occupationally exposed to microwave radiation

Lowering the quantification limit of the QubitTM RNA HS assay using RNA spike-in.

Science.gov (United States)

Li, Xin; Ben-Dov, Iddo Z; Mauro, Maurizio; Williams, Zev

2015-05-06

RNA quantification is often a prerequisite for most RNA analyses such as RNA sequencing. However, the relatively low sensitivity and large sample consumption of traditional RNA quantification methods such as UV spectrophotometry and even the much more sensitive fluorescence-based RNA quantification assays, such as the Qubit™ RNA HS Assay, are often inadequate for measuring minute levels of RNA isolated from limited cell and tissue samples and biofluids. Thus, there is a pressing need for a more sensitive method to reliably and robustly detect trace levels of RNA without interference from DNA. To improve the quantification limit of the Qubit™ RNA HS Assay, we spiked-in a known quantity of RNA to achieve the minimum reading required by the assay. Samples containing trace amounts of RNA were then added to the spike-in and measured as a reading increase over RNA spike-in baseline. We determined the accuracy and precision of reading increases between 1 and 20 pg/μL as well as RNA-specificity in this range, and compared to those of RiboGreen(®), another sensitive fluorescence-based RNA quantification assay. We then applied Qubit™ Assay with RNA spike-in to quantify plasma RNA samples. RNA spike-in improved the quantification limit of the Qubit™ RNA HS Assay 5-fold, from 25 pg/μL down to 5 pg/μL while maintaining high specificity to RNA. This enabled quantification of RNA with original concentration as low as 55.6 pg/μL compared to 250 pg/μL for the standard assay and decreased sample consumption from 5 to 1 ng. Plasma RNA samples that were not measurable by the Qubit™ RNA HS Assay were measurable by our modified method. The Qubit™ RNA HS Assay with RNA spike-in is able to quantify RNA with high specificity at 5-fold lower concentration and uses 5-fold less sample quantity than the standard Qubit™ Assay.

The modified nanocrystalline cellulose for hydrophobic drug delivery

Energy Technology Data Exchange (ETDEWEB)

Qing, Weixia [Institute of Environmental and Analytical Sciences, College of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004 (China); Medical College, Henan University, Kaifeng 475004 (China); Wang, Yong [Institute of Environmental and Analytical Sciences, College of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004 (China); Wang, Youyou [Institute of Environmental and Analytical Sciences, College of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004 (China); Key Lab of Natural Medicine and Immun-engineering of Henan Province, Henan University, Kaifeng 475004 (China); Zhao, Dongbao [Institute of Environmental and Analytical Sciences, College of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004 (China); Liu, Xiuhua, E-mail: ll514527@163.com [Institute of Environmental and Analytical Sciences, College of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004 (China); Key Lab of Natural Medicine and Immun-engineering of Henan Province, Henan University, Kaifeng 475004 (China); Zhu, Jinhua [Institute of Environmental and Analytical Sciences, College of Chemistry and Chemical Engineering, Henan University, Kaifeng 475004 (China)

2016-03-15

Graphical abstract: - Highlights: • Torispherical NCC was synthesized through the improvements on the hydrolysis method. • NCC was firstly modified with CTMAB as a drug carrier. • Luteolin and luteoloside loading CTMAB-coated NCC were studied. - Abstract: In this work, torispherical nanocrystalline cellulose (NCC) was synthesized, and firstly modified with a cationic surfactant cetyltrimethylammonium bromide (CTMAB). It was proved that the kinetics of NCC adsorbing CTMAB followed the pseudo-second-order kinetics equation, and the adsorption isotherm model followed Freundlich which was multi molecular layer adsorption model. The morphology and structure of NCC and CTMAB-coated NCC were characterized by transmission electron microscopy (TEM) and X-ray powder diffraction (XRD). Stabilities of NCC and CTMAB-coated NCC were assayed by zeta potential. The results showed that NCC in CTMAB solution was well-dispersed and stable. Moreover, the drug loading and release performance of CTMAB-coated NCC were studied using luteolin (LUT) and luteoloside (LUS) as model drugs.

The modified nanocrystalline cellulose for hydrophobic drug delivery

International Nuclear Information System (INIS)

Qing, Weixia; Wang, Yong; Wang, Youyou; Zhao, Dongbao; Liu, Xiuhua; Zhu, Jinhua

2016-01-01

Graphical abstract: - Highlights: • Torispherical NCC was synthesized through the improvements on the hydrolysis method. • NCC was firstly modified with CTMAB as a drug carrier. • Luteolin and luteoloside loading CTMAB-coated NCC were studied. - Abstract: In this work, torispherical nanocrystalline cellulose (NCC) was synthesized, and firstly modified with a cationic surfactant cetyltrimethylammonium bromide (CTMAB). It was proved that the kinetics of NCC adsorbing CTMAB followed the pseudo-second-order kinetics equation, and the adsorption isotherm model followed Freundlich which was multi molecular layer adsorption model. The morphology and structure of NCC and CTMAB-coated NCC were characterized by transmission electron microscopy (TEM) and X-ray powder diffraction (XRD). Stabilities of NCC and CTMAB-coated NCC were assayed by zeta potential. The results showed that NCC in CTMAB solution was well-dispersed and stable. Moreover, the drug loading and release performance of CTMAB-coated NCC were studied using luteolin (LUT) and luteoloside (LUS) as model drugs.

Evaluation of four endogenous reference genes and their real-time PCR assays for common wheat quantification in GMOs detection.

Science.gov (United States)

Huang, Huali; Cheng, Fang; Wang, Ruoan; Zhang, Dabing; Yang, Litao

2013-01-01

Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs) detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L.) DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat.

Evaluation of four endogenous reference genes and their real-time PCR assays for common wheat quantification in GMOs detection.

Directory of Open Access Journals (Sweden)

Huali Huang

Full Text Available Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L. DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat.

Evaluation of Four Endogenous Reference Genes and Their Real-Time PCR Assays for Common Wheat Quantification in GMOs Detection

Science.gov (United States)

Huang, Huali; Cheng, Fang; Wang, Ruoan; Zhang, Dabing; Yang, Litao

2013-01-01

Proper selection of endogenous reference genes and their real-time PCR assays is quite important in genetically modified organisms (GMOs) detection. To find a suitable endogenous reference gene and its real-time PCR assay for common wheat (Triticum aestivum L.) DNA content or copy number quantification, four previously reported wheat endogenous reference genes and their real-time PCR assays were comprehensively evaluated for the target gene sequence variation and their real-time PCR performance among 37 common wheat lines. Three SNPs were observed in the PKABA1 and ALMT1 genes, and these SNPs significantly decreased the efficiency of real-time PCR amplification. GeNorm analysis of the real-time PCR performance of each gene among common wheat lines showed that the Waxy-D1 assay had the lowest M values with the best stability among all tested lines. All results indicated that the Waxy-D1 gene and its real-time PCR assay were most suitable to be used as an endogenous reference gene for common wheat DNA content quantification. The validated Waxy-D1 gene assay will be useful in establishing accurate and creditable qualitative and quantitative PCR analysis of GM wheat. PMID:24098735

Development of a biomimetic enzyme-linked immunosorbent assay based on molecularly imprinted polymers on paper for the detection of carbaryl.

Science.gov (United States)

Zhang, Can; Cui, Hanyu; Han, Yufeng; Yu, Fangfang; Shi, Xiaoman

2018-02-01

A biomimetic enzyme-linked immunosorbent assay (BELISA) which was based on molecularly imprinted polymers on paper (MIPs-paper) with specific recognition was developed. As a detector, the surface of paper was modified with γ-MAPS by hydrolytic action and anchored the MIP layer on γ-MAPS modified-paper by copolymerization to construct the artificial antibody Through a series of experimentation and verification, we successful got the MIPs-paper and established BELISA for the detection of carbaryl. The development of MIPs-paper based on BELISA was applied to detect carbaryl in real samples and validated by an enzyme-linked immunosorbent assay (ELISA) based on anti-carbaryl biological antibody. The results of these two methods (BELISA and ELISA) were well correlated (R 2 =0.944). The established method of MIPs-paper BELISA exhibits the advantages of low cost, higher stability and being re-generable, which can be applied as a convenient tool for the fast and efficient detection of carbaryl. Copyright © 2017. Published by Elsevier Ltd.

Assay strategies and methods for phospholipases

International Nuclear Information System (INIS)

Reynolds, L.J.; Washburn, W.N.; Deems, R.A.; Dennis, E.A.

1991-01-01

Of the general considerations discussed, the two issues which are most important in choosing an assay are (1) what sensitivity is required to assay a particular enzyme and (2) whether the assay must be continuous. One can narrow the options further by considering substrate availability, enzyme specificity, assay convenience, or the presence of incompatible side reactions. In addition, the specific preference of a particular phospholipase for polar head group, micellar versus vesicular substrates, and anionic versus nonionic detergents may further restrict the options. Of the many assays described in this chapter, several have limited applicability or serious drawbacks and are not commonly employed. The most commonly used phospholipase assays are the radioactive TLC assay and the pH-stat assay. The TLC assay is probably the most accurate, sensitive assay available. These aspects often outweigh the disadvantages of being discontinuous, tedious, and expensive. The radioactive E. coli assay has become popular recently as an alternative to the TLC assay for the purification of the mammalian nonpancreatic phospholipases. The assay is less time consuming and less expensive than the TLC assay, but it is not appropriate when careful kinetics are required. Where less sensitivity is needed, or when a continuous assay is necessary, the pH-stat assay is often employed. With purified enzymes, when free thiol groups are not present, a spectrophotometric thiol assay can be used. This assay is ∼ as sensitive as the pH-stat assay but is more convenient and more reproducible, although the substrate is not available commercially. Despite the many assay choices available, the search continues for a convenient, generally applicable assay that is both sensitive and continuous

Synthesis of photothermal nanocomposites and their application to antibacterial assays

Science.gov (United States)

Yang, Ning; Wang, Chun; Wang, Xiaoyu; Li, Lidong

2018-04-01

In this work, we report a novel gold nanorod (AuNR)-based nanocomposite that shows strong binding to bacterium and high antibacterial efficiency. The AuNRs were used as a photothermal material to transform near-infrared radiation (NIR) into heat. We selected poly (acrylic acid) to modify the surface of the AuNRs based on a simple self-assembly method. After conjugation of the bacterium-binding molecule vancomycin, the nanocomposites were capable of efficiently gathering on the cell walls of bacteria. The nanocomposites exhibited a high bacterial inhibition capability owing to NIR-induced heat generation in situ. Therefore, the prepared photothermal nanocomposites show great potential for use in antibacterial assays.

Solid phase assays

International Nuclear Information System (INIS)

Reese, M.G.; Johnson, L.R.; Ransom, D.K.

1980-01-01

In a solid phase assay for quantitative determination of biological and other analytes, a sample such as serum is contacted with a receptor for the analyte being assayed, the receptor being supported on a solid support. No tracer for the analyte is added to the sample before contacting with the receptor; instead the tracer is contacted with the receptor after unbound analyte has been removed from the receptor. The assay can be otherwise performed in a conventional manner but can give greater sensitivity. (author)

Detection of induced male germline mutation: Correlations and comparisons between traditional germline mutation assays, transgenic rodent assays and expanded simple tandem repeat instability assays

Energy Technology Data Exchange (ETDEWEB)

Singer, Timothy M. [Mutagenesis Section, Environmental and Occupational Toxicology Division, Safe Environments Programme, 0803A, Health Canada, Ottawa, Ont., K1A 0K9 (Canada); Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, Ont., K1S 5B6 (Canada); Lambert, Iain B. [Department of Biology, Carleton University, 1125 Colonel By Drive, Ottawa, Ont., K1S 5B6 (Canada); Williams, Andrew [Biostatistics and Epidemiology Division, Safe Environments Programme, 6604B, Health Canada, Ottawa, Ont., K1A 0K9 (Canada); Douglas, George R. [Mutagenesis Section, Environmental and Occupational Toxicology Division, Safe Environments Programme, 0803A, Health Canada, Ottawa, Ont., K1A 0K9 (Canada); Yauk, Carole L. [Mutagenesis Section, Environmental and Occupational Toxicology Division, Safe Environments Programme, 0803A, Health Canada, Ottawa, Ont., K1A 0K9 (Canada)]. E-mail: carole_yauk@hc-sc.gc.ca

2006-06-25

Several rodent assays are capable of monitoring germline mutation. These include traditional assays, such as the dominant lethal (DL) assay, the morphological specific locus (SL) test and the heritable translocation (HT) assay, and two assays that have been developed more recently-the expanded simple tandem repeat (ESTR) and transgenic rodent (TGR) mutation assays. In this paper, we have compiled the limited amount of experimental data that are currently available to make conclusions regarding the comparative ability of the more recently developed assays to detect germline mutations induced by chemical and radiological agents. The data suggest that ESTR and TGR assays are generally comparable with SL in detecting germline mutagenicity induced by alkylating agents and radiation, though TGR offered less sensitivity than ESTR in some cases. The DL and HT assays detect clastogenic events and are most susceptible to mutations arising in post-spermatogonial cells, and they may not provide the best comparisons with TGR and ESTR instability. The measurement of induced ESTR instability represents a relatively sensitive method of identifying agents causing germline mutation in rodents, and may also be useful for bio-monitoring exposed individuals in the human population. Any future use of the TGR and ESTR germline mutation assays in a regulatory testing context will entail more robust and extensive characterization of assay performance. This will require substantially more data, including experiments measuring multiple endpoints, a greatly expanded database of chemical agents and a focus on characterizing stage-specific activity of mutagens in these assays, preferably by sampling epididymal sperm exposed at defined pre-meiotic, meiotic and post-meiotic stages of development.

Detection of induced male germline mutation: Correlations and comparisons between traditional germline mutation assays, transgenic rodent assays and expanded simple tandem repeat instability assays

International Nuclear Information System (INIS)

Singer, Timothy M.; Lambert, Iain B.; Williams, Andrew; Douglas, George R.; Yauk, Carole L.

2006-01-01

Several rodent assays are capable of monitoring germline mutation. These include traditional assays, such as the dominant lethal (DL) assay, the morphological specific locus (SL) test and the heritable translocation (HT) assay, and two assays that have been developed more recently-the expanded simple tandem repeat (ESTR) and transgenic rodent (TGR) mutation assays. In this paper, we have compiled the limited amount of experimental data that are currently available to make conclusions regarding the comparative ability of the more recently developed assays to detect germline mutations induced by chemical and radiological agents. The data suggest that ESTR and TGR assays are generally comparable with SL in detecting germline mutagenicity induced by alkylating agents and radiation, though TGR offered less sensitivity than ESTR in some cases. The DL and HT assays detect clastogenic events and are most susceptible to mutations arising in post-spermatogonial cells, and they may not provide the best comparisons with TGR and ESTR instability. The measurement of induced ESTR instability represents a relatively sensitive method of identifying agents causing germline mutation in rodents, and may also be useful for bio-monitoring exposed individuals in the human population. Any future use of the TGR and ESTR germline mutation assays in a regulatory testing context will entail more robust and extensive characterization of assay performance. This will require substantially more data, including experiments measuring multiple endpoints, a greatly expanded database of chemical agents and a focus on characterizing stage-specific activity of mutagens in these assays, preferably by sampling epididymal sperm exposed at defined pre-meiotic, meiotic and post-meiotic stages of development

"Life-like" assessment of antimicrobial surfaces by a new touch transfer assay displays strong superiority of a copper alloy compared to silver containing surfaces.

Directory of Open Access Journals (Sweden)

Johannes Karl-Mark Knobloch

Full Text Available Transmission of bacteria from inanimate surfaces in healthcare associated environments is an important source of hospital acquired infections. A number of commercially available medical devices promise to fulfill antibacterial activity to reduce environmental contamination. In this study we developed a touch transfer assay modeling fingerprint transmission to investigate the antibacterial activity of surfaces, with confirmed antibacterial activity by a modified ISO 22196 (JIS Z 2801 assay to test such surfaces under more realistic conditions. Bacteria were taken up from a dry standardized primary contaminated surface (PCS with disinfected fingers or fingers covered with sterile and moistened cotton gloves. Subsequently, bacteria were transferred by pressing on secondary contaminated surfaces (SCS with or without potential antibacterial activity and the relative reduction rate was determined after 24 h. A stable transmission rate between PCS and SCS was observed using moistened sterile gloves. A copper containing alloy displayed at least a tenfold reduction of the bacterial load consistently reaching less than 2.5 cfu/cm2. In contrast, no significant reduction of bacterial contamination by silver containing surfaces and matured pure silver was observed in the touch transfer assay. With the touch transfer assay we successfully established a new reproducible method modeling cross contamination. Using the new method we were able to demonstrate that several surfaces with confirmed antimicrobial activity in a modified ISO 22196 (JIS Z 2801 assay lacked effectiveness under defined ambient conditions. This data indicate that liquid based assays like the ISO 22196 should be critically reviewed before claiming antibacterial activity for surfaces in the setting of contamination of dry surfaces by contact to the human skin. We suggest the newly developed touch transfer assay as a new additional tool for the assessment of potential antimicrobial surfaces

Effects of Multivitamins and Known Teratogens on Chick Cardiomyocytes Micromass Culture Assay

Directory of Open Access Journals (Sweden)

Samreen Memon

2013-09-01

Full Text Available   Objective(s: This study aimed to find out whether the chick cardiomyocyte micromass (MM system could be employed to predict the teratogenecity of common environmental factors. Different multivitamins and over the counter drugs were used in this study.   Materials and Methods: White Leghorn 5-day-old embryo hearts were dissected and trypsinized to produce a cardiomyocyte cell suspension in Dulbecco's Modified Eagle's Medium. The cultures were incubated at 370C in 5% CO2 in air, and observations were made at 24, 48 and 144 hr, for the detection of cell beating. Cellular viability was assessed using the resazurin assay and cell protein content was assessed by the kenacid blue assay. It was observed that while not affecting total cell number folic acid, vitamin C, sodium fluoride and ginseng did not significantly reduced cell activity and beating. However cadmium chloride significantly reduced the beating, cell viability and cell protein content in micromass cultures. Results: The results demonstrate the potential of the chick cardiomyocyte MM culture assay to identify teratogens/embryotoxins that alter morphology and function, which may result in either teratogenic outcome or cytotoxicity. Conclusion: This could form part of a screen for developmental toxicity related to cardiac function

Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

International Nuclear Information System (INIS)

Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro; Onodera, Takeshi; Toko, Kiyoshi

2014-01-01

The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

Development of Indirect Competitive Immuno-Assay Method Using SPR Detection for Rapid and Highly Sensitive Measurement of Salivary Cortisol Levels

Energy Technology Data Exchange (ETDEWEB)

Tahara, Yusuke; Huang, Zhe; Kiritoshi, Tetsuro [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Onodera, Takeshi [Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan); Toko, Kiyoshi, E-mail: toko@ed.kyushu-u.ac.jp [Graduate School of Information Science and Electrical Engineering, Kyushu University, Fukuoka (Japan); Research and Development Center for Taste and Odor Sensing, Kyushu University, Fukuoka (Japan)

2014-05-30

The monitoring of salivary cortisol as a key biomarker of an individual’s stress response has been increasingly focused on. This paper describes the development of a novel cortisol immuno-assay method based on an indirect competitive method using a commercially available surface plasmon resonance instrument. The surface of an Au chip was modified with PEG6-COOH aromatic dialkanethiol self-assembled monolayers and hydrocortisone 3-(O-carboxymethyl) oxime (hydrocortisone 3-CMO) as a cortisol analog. A detection limit of 38 ppt range with a measurement range of 10 ppt–100 ppb was accomplished without the incubation of a mixing solution consisting of standard cortisol and an anti-cortisol antibody, and the time for quantification of cortisol concentration was 8 min from the sample injection. We experimentally compared our immuno-assay with a commercialized salivary cortisol enzyme-linked immunosorbent assay (ELISA) kit using human saliva samples. It was found that the results obtained by the cortisol immuno-assay had a good correlation with those obtained by ELISA assay (R = 0.96). Our findings indicate the potential utility of the cortisol immuno-assay for measurements of human salivary cortisol levels.

Evaluation of total PSA assay on vitros ECi and correlation with Kryptor-PSA assay.

Science.gov (United States)

Cassinat, B; Wacquet, M; Toubert, M E; Rain, J D; Schlageter, M H

2001-01-01

An increasing number of multiparametric immuno-analysers for PSA assays are available. As different immuno-assays may vary in their analytical quality and their accuracy for the follow-up of patients, expertise is necessary for each new assay. The PSA assay on the Vitros-ECi analyser has been evaluated and compared with the PSA assay from the Kryptor analyser. Variation coefficients were 0.91 to 1.98% for within-run assays, and 4.2% to 5.4% for interassay (PSA levels = 0.8 microgram/L to 33.6 micrograms/L). Dilution tests showed 93 to 136% recovery until 70 micrograms/L PSA. Functional sensitivity was estimated at 0.03 microgram/L. Equimolarity of the test was confirmed. Correlation of PSA levels measured with Vitros-ECi and Kryptor analysers displayed a correlation coefficient r2 of 0.9716. The half-lives and doubling times of PSA were similar using both methods. Vitros-ECi PSA assay meets the major criteria for the management of prostate cancer patients.

Hormone assay

International Nuclear Information System (INIS)

Eisentraut, A.M.

1977-01-01

An improved radioimmunoassay is described for measuring total triiodothyronine or total thyroxine levels in a sample of serum containing free endogenous thyroid hormone and endogenous thyroid hormone bound to thyroid hormone binding protein. The thyroid hormone is released from the protein by adding hydrochloric acid to the serum. The pH of the separated thyroid hormone and thyroid hormone binding protein is raised in the absence of a blocking agent without interference from the endogenous protein. 125 I-labelled thyroid hormone and thyroid hormone antibodies are added to the mixture, allowing the labelled and unlabelled thyroid hormone and the thyroid hormone antibody to bind competitively. This results in free thyroid hormone being separated from antibody bound thyroid hormone and thus the unknown quantity of thyroid hormone may be determined. A thyroid hormone test assay kit is described for this radioimmunoassay. It provides a 'single tube' assay which does not require blocking agents for endogenous protein interference nor an external solid phase sorption step for the separation of bound and free hormone after the competitive binding step; it also requires a minimum number of manipulative steps. Examples of the assay are given to illustrate the reproducibility, linearity and specificity of the assay. (UK)

A Rapid Method for Quantifying Viable Mycobacterium avium subsp. paratuberculosis in Cellular Infection Assays

Science.gov (United States)

Pooley, Hannah B.; de Silva, Kumudika; Purdie, Auriol C.; Begg, Douglas J.; Whittington, Richard J.

2016-01-01

ABSTRACT Determining the viability of bacteria is a key outcome of in vitro cellular infection assays. Currently, this is done by culture, which is problematic for fastidious slow-growing bacteria such as Mycobacterium avium subsp. paratuberculosis, where it can take up to 4 months to confirm growth. This study aimed to identify an assay that can rapidly quantify the number of viable M. avium subsp. paratuberculosis cells in a cellular sample. Three commercially available bacterial viability assays along with a modified liquid culture method coupled with high-throughput quantitative PCR growth detection were assessed. Criteria for assessment included the ability of each assay to differentiate live and dead M. avium subsp. paratuberculosis organisms and their accuracy at low bacterial concentrations. Using the culture-based method, M. avium subsp. paratuberculosis growth was reliably detected and quantified within 2 weeks. There was a strong linear association between the 2-week growth rate and the initial inoculum concentration. The number of viable M. avium subsp. paratuberculosis cells in an unknown sample was quantified based on the growth rate, by using growth standards. In contrast, none of the commercially available viability assays were suitable for use with samples from in vitro cellular infection assays. IMPORTANCE Rapid quantification of the viability of Mycobacterium avium subsp. paratuberculosis in samples from in vitro cellular infection assays is important, as it allows these assays to be carried out on a large scale. In vitro cellular infection assays can function as a preliminary screening tool, for vaccine development or antimicrobial screening, and also to extend findings derived from experimental animal trials. Currently, by using culture, it takes up to 4 months to obtain quantifiable results regarding M. avium subsp. paratuberculosis viability after an in vitro infection assay; however, with the quantitative PCR and liquid culture method

Development and validation of a 48-target analytical method for high-throughput monitoring of genetically modified organisms.

Science.gov (United States)

Li, Xiaofei; Wu, Yuhua; Li, Jun; Li, Yunjing; Long, Likun; Li, Feiwu; Wu, Gang

2015-01-05

The rapid increase in the number of genetically modified (GM) varieties has led to a demand for high-throughput methods to detect genetically modified organisms (GMOs). We describe a new dynamic array-based high throughput method to simultaneously detect 48 targets in 48 samples on a Fludigm system. The test targets included species-specific genes, common screening elements, most of the Chinese-approved GM events, and several unapproved events. The 48 TaqMan assays successfully amplified products from both single-event samples and complex samples with a GMO DNA amount of 0.05 ng, and displayed high specificity. To improve the sensitivity of detection, a preamplification step for 48 pooled targets was added to enrich the amount of template before performing dynamic chip assays. This dynamic chip-based method allowed the synchronous high-throughput detection of multiple targets in multiple samples. Thus, it represents an efficient, qualitative method for GMO multi-detection.

Dose assessment of SiC nanoparticle dispersions during in vitro assays

International Nuclear Information System (INIS)

Mejia, Jorge; Piret, Jean-Pascal; Noël, Florence; Masereel, Bernard; Toussaint, Olivier; Lucas, Stéphane

2013-01-01

Here, we show that key physicochemical parameters of commercial Silicon Carbide nanoparticles, such as the primary particles of about 53 nm in size, the agglomerates size, and the surface composition, are considerably modified with respect to the pristine conditions, during in vitro assessment. The use of sample conditioning stages, such as the pre-dispersion in aqueous media and the subsequent dispersion in a culture medium specific to the in vitro assay, produce modifications as the absorption of N, C, and O, from the culture medium, in the nanoparticles surface. Our results show that the sedimented dose, fraction of sedimented NPs during incubation and consequently in contact with cells seeded at the bottom, of Silicon Carbide nanoparticles can be measured from the particle size distribution obtained using a centrifugal liquid sedimentation technique. It is underlined that the variations observed in the physicochemical properties are related to the in vitro assay conditions. Culture medium and incubation time are found to influence the most the sedimented dose and consequently the cells dose uptake

Ex vivo assays to study self-renewal, long-term expansion, and leukemic transformation of genetically modified human hematopoietic and patient-derived leukemic stem cells

NARCIS (Netherlands)

Sontakke, Pallavi; Carretta, Marco; Capala, Marta; Schepers, Hein; Schuringa, Jan Jacob

2014-01-01

With the emergence of the concept of the leukemic stem cell (LSC), assays to study them remain pivotal in understanding (leukemic) stem cell biology. Although the in vivo NOD-SCID or NSG xenotransplantation model is currently still the favored assay of choice in most cases, this system has some

Reversible control of kinesin activity and microtubule gliding speeds by switching the doping states of a conducting polymer support

Energy Technology Data Exchange (ETDEWEB)

Martin, Brett D [US Naval Research Laboratory, Code 6930, Washington, DC 20375 (United States); Velea, Luminita M [US Naval Research Laboratory, Code 6930, Washington, DC 20375 (United States); Soto, Carissa M [US Naval Research Laboratory, Code 6930, Washington, DC 20375 (United States); Whitaker, Craig M [US Naval Academy, Department of Chemistry, Annapolis, MD 21402 (United States); Gaber, Bruce P [US Naval Research Laboratory, Code 6930, Washington, DC 20375 (United States); Ratna, Banahalli [US Naval Research Laboratory, Code 6930, Washington, DC 20375 (United States)

2007-02-07

We describe a method for reversibly controlling the ATPase activity of streptavidin-linked kinesin by changing the doping states of a conducting polymer support. When the polymer (poly(CH{sub 2}OH-EDOT)) was electrochemically switched from its dedoped (semiconducting) state to its doped (conducting) state, the ATPase activity of the adsorbed kinesin complex decreased by 35% with a concomitant decrease in the gliding speeds of kinesin-driven microtubules. When the polymer was switched back to its original dedoped state, nearly identical increases were observed in the kinesin ATPase activity and microtubule speeds. Use of a fluorescent ATP substrate analogue showed that the total amount of kinesin adsorbed on the poly(CH{sub 2}OH-EDOT) surface remained constant as the doping state of the polymer was switched. The microtubules exhibited nearly identical speed differences on the doped and dedoped surfaces for both chemical and electrochemical doping methods. Michaelis-Menten modelling suggests that the doped surface acts as an 'uncompetitive inhibitor' of kinesin. This work represents an investigation into the phenomenon of an electrically switchable surface exerting a moderating effect on the activity of an adsorbed protein that does not contain a bound, electroactive metal ion.

Biochemical studies of mouse brain tubulin: colchicine binding (DEAE-cellulose filter) assay and subunits (α and β) biosynthesis and degradation (in newborn brain)

International Nuclear Information System (INIS)

Tse, C.F.

1978-01-01

A DEAE-cellulose filter assay, measuring [ 3 H]colchicine bound to colchicine binding protein (CBP) absorbed on filter discs, has been modified to include lM sucrose in the incubation medium for complexing colchicine to CBP in samples before applying the samples to filter discs (single point assay). Due to the much greater stability of colchicine binding capacity in the presence of lM sucrose, multiple time-point assays and least squares linear regression analysis were not necessary for accurate determination of CBP in hybrid mouse brain at different stages of development. The highest concentrations of CBP were observed in the 160,000g supernatant and pellet of newborn brain homogenate. Further studies of the modified filter assay documented that the assay has an overall counting efficiency of 27.3%, that DEAE-cellulose filters bind and retain all tubulin in the assay samples, and that one molecule of colchicine binds approximately one molecule of tubulin dimer. Therefore, millimoles of colchicine bound per milligram total protein can be used to calculate tubulin content. With this technique tubulin content of brain supernatant was found to be 11.9% for newborn, and 7.15% for 11 month old mice. Quantitative densitometry was also used to measure mouse brain supernatant actin content for these two stages. In vivo synthesis and degradation rates of tubulin α and β subunits of two day mouse brain 100,000g supernatant were studied after intracerebral injection of [ 3 H]leucine. Quantitative changes of the ratio of tritium specific activities of tubulin α and β subunits with time were determined. The pattern of change was biphasic. During the first phase the ratio decreased; during the second phase the ratio increased continuously. An interpretation consistent with all the data in this study is that the α subunit is synthesized at a more rapid rate than the β subunit

Colorimetric and ratiometric aggregation assay for streptomycin using gold nanoparticles and a new and highly specific aptamer

International Nuclear Information System (INIS)

Soheili, Vahid; Taghdisi, Seyed Mohammad; Khayyat, Mohammad Hassanzadeh; Abnous, Khalil; Bazzaz, BiBi Sedigheh Fazly; Ramezani, Mohammad

2016-01-01

Aptamers specific for the antibiotic streptomycin were identified by a modified SELEX procedure that employs magnetic beads. After eight rounds of selection, twenty-six aptamers were identified and clustered into seven groups according to similarities in their sequences. The binding constant of three sequences from different groups were determined by colorimetric assays using unmodified gold nanoparticles (AuNPs). These most suitable aptamers were then truncated, and finally a 23-base sequence was identified that has the highest affinity (K_d = 132.3 nM) and selectivity. The assay was employed to analyze streptomycin residue in raw milk samples by ratiometric spectrophotometry at 520 and 660 nm, respectively. The analytical range extends from 180 to 1000 nM, and the LOD is 47.2 nM which is better than that of HPLC (4 μM). The interaction between aptamer and streptomycin was studied by molecular modeling. In our perception, this colorimetric assay provides a viable method for fast analysis of streptomycin in raw milk. (author)

A modified spectrophotometric assay to estimate deglycosylation of steroidal saponin to sapogenin by mixed ruminal microbes.

Science.gov (United States)

Wang, Yuxi; McAllister, Tim A

2010-08-30

The lack of a method for measuring deglycosylation of saponins in ruminal fluid has limited the ability to investigate the impact of these compounds on rumen microorganisms. A simple spectrophotometric assay was adapted and a protocol developed to enable measurement of steroidal saponin and sapogenin in ruminal fluid. The procedure was used for in vitro determination of deglycosylation activity of rumen bacteria obtained from cattle fed or not fed Yucca schidigera saponin, and to determine the relative deglycosylase activities of extracellular and cell-associated enzymes from ruminal content. Modifications to the spectrophotometric assay (i.e. heating time shortened to 10 min and 0.5 mL dH(2)O added to the reaction mixture) improved the stability of the optical density (425 nm) of the chromophore for up to 24 h post-reaction. Centrifugation (12 000 x g, 20 min) enabled differential estimations of steroidal saponin and sapogenin in ruminal fluid. Steroidal saponin added to defaunated ruminal fluid (dRF) or clarified ruminal fluid (cRF) was recovered completely from the mixture as saponin + sapogenin (99.1% and 100.6%, respectively), whereas saponin recovery from the supernatant of dRF was greatly reduced (P ruminal extracellular enzyme preparation was recoverable in supernatant after 24 h, compared with only 26-32% remaining in supernatant from incubation with a cell-associated enzymes fraction. Mixed rumen bacteria deglycosylate steroidal saponin to sapogenin, at activity levels unaffected by prior exposure to saponin, but they were unable to degrade the sapogenin core structure. Deglycosylation activity occurred primarily in the cell-associated enzyme fraction of ruminal content. Copyright (c) 2008 Crown in the right of Canada. Published by John Wiley & Sons, Ltd.

The Comet Assay: Tails of the (Unexpected. Use of the comet assay in pharmaceutical development.

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Bas-jan Van Der Leede

2015-08-01

Full Text Available In genotoxicity testing of pharmaceuticals the rodent alkaline comet assay is being increasingly used as a second in vivo assay in addition to the in vivo micronucleus assay to mitigate in vitro positive results as recommended by regulatory guidance. In this presentation we want to give insight into the circumstances in vivo comet assay is deployed in a Genetic Toxicology Department of a pharmaceutical company. As the in vivo comet assay is a salvage assay, it means that some events have occurred in an in vitro assay and that the compound (or metabolite responsible for this signal is potentially deselected for further development. More than often the decision to perform an in vivo comet assay is at a very early stage in development and the first time that the compound will be tested in vivo at high/toxic dose levels. As almost no toxicokinetic data and tissue distribution data are available a careful design with maximizes the chances for successful mitigation is necessary. Decisions on acute or repeated dosing need to be made and arrangements for combining the in vivo comet assay with the in vivo micronucleus assay are to be considered. Often synthesis methods need to be scaled up fast to provide the required amount of compound and information on suitable formulations needs to be in place. As exposure data is crucial for interpretation of results, analytical methods need to be brought in place rapidly. An experienced multi skilled and communicative team needs to be available to deploy successfully this kind of assays at an early stage of development. We will present a few scenarios on study conduct and demonstrate how this assay can make a difference for the further development of a new drug.

Application of the KeratinoSens™ assay for assessing the skin sensitization potential of agrochemical active ingredients and formulations.

Science.gov (United States)

Settivari, Raja S; Gehen, Sean C; Amado, Ricardo Acosta; Visconti, Nicolo R; Boverhof, Darrell R; Carney, Edward W

2015-07-01

Assessment of skin sensitization potential is an important component of the safety evaluation process for agrochemical products. Recently, non-animal approaches including the KeratinoSens™ assay have been developed for predicting skin sensitization potential. Assessing the utility of the KeratinoSens™ assay for use with multi-component mixtures such as agrochemical formulations has not been previously evaluated and is a significant need. This study was undertaken to evaluate the KeratinoSens™ assay prediction potential for agrochemical formulations. The assay was conducted for 8 agrochemical active ingredients (AIs) including 3 sensitizers (acetochlor, meptyldinocap, triclopyr), 5 non-sensitizers (aminopyralid, clopyralid, florasulam, methoxyfenozide, oxyfluorfen) and 10 formulations for which in vivo sensitization data were available. The KeratinoSens™ correctly predicted the sensitization potential of all the AIs. For agrochemical formulations it was necessary to modify the standard assay procedure whereby the formulation was assumed to have a common molecular weight. The resultant approach correctly predicted the sensitization potential for 3 of 4 sensitizing formulations and all 6 non-sensitizing formulations when compared to in vivo data. Only the meptyldinocap-containing formulation was misclassified, as a result of high cytotoxicity. These results demonstrate the promising utility of the KeratinoSens™ assay for evaluating the skin sensitization potential of agrochemical AIs and formulations. Copyright © 2015 Elsevier Inc. All rights reserved.

Studies on the utility and mechanism of the V-79 cell metabolic cooperation assay for tumor promoters

International Nuclear Information System (INIS)

Hartman, T.G.

1985-01-01

Cigarette smoke condensate and its fractions were tested for activity in the V-79 Metabolic Cooperation Assay to determine the usefulness of the assay for analysis of a complex mixture and to compare the results obtained with previously conducted in vivo promoter assays. The whole condensate and several of its fractions were positive in the assay. In general, the Metabolic Cooperation Assay results were comparable to previously published results obtained on mouse skin. The effect of cell density, phorbol 12-myrystate-13-acetate (PMA) exposure time, concentration, pre-exposure and binding activity on the recovery of mutant V-79 Chinese hamster lung fibroblasts in the Metabolic Cooperation Assay was determined. A PMA exposure interval of only 1 minute resulted in maximum recovery of mutant cells. PMA began to inhibit metabolic cooperation at an exposure concentration of 0.1 ng/ml. Pre-exposure of cells to PMA increased the recovery of both post-PMA-treated and non-treated mutant cells in a dose-dependent manner. 3 H-PMA was rapidly bound to or taken up by the V-79 cells under assay conditions. The effect of calcium antagonists and representative compounds from several classes of anti-promoters including anti-inflammatory sterols, protease inhibitors, retinoids and cyclic nucleotides on metabolic determined. Each compound was tested for its effect on metabolic cooperation and also for its ability to reverse or modify the inhibitory properties of PMA on inter-cellular communication. Of all the compounds tested only cyclic adenosine monophosphate (cAMP) was able to antagonize the inhibitory effect of PMA

A novel assay of DGAT activity based on high temperature GC/MS of triacylglycerol.

Science.gov (United States)

Greer, Michael S; Zhou, Ting; Weselake, Randall J

2014-08-01

Diacylglycerol acyltransferase (DGAT) catalyzes the final step in the acyl-CoA-dependent biosynthesis of triacylglycerol (TAG), a high-energy compound composed of three fatty acids esterified to a glycerol backbone. In vitro DGAT assays, which are usually conducted with radiolabeled substrate using microsomal fractions, have been useful in identifying compounds and genetic modifications that affect DGAT activity. Here, we describe a high-temperature gas chromatography (GC)/mass spectrometry (MS)-based method for monitoring molecular species of TAG produced by the catalytic action of microsomal DGAT. This method circumvents the need for radiolabeled or modified substrates, and only requires a simple lipid extraction prior to GC. The utility of the method is demonstrated using a recombinant type-1 Brassica napus DGAT produced in a strain of Saccharomyces cerevisae that is deficient in TAG synthesis. The GC/MS-based assay of DGAT activity was strongly correlated with the typical in vitro assay of the enzyme using [1-(14)C] acyl-CoA as an acyl donor. In addition to determining DGAT activity, the method is also useful for determining substrate specificity and selectivity properties of the enzyme.

Population screening for glucose-6-phosphate dehydrogenase deficiencies in Isabel Province, Solomon Islands, using a modified enzyme assay on filter paper dried bloodspots

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Landry Losi

2010-08-01

Full Text Available Abstract Background Glucose-6-phosphate dehydrogenase deficiency poses a significant impediment to primaquine use for the elimination of liver stage infection with Plasmodium vivax and for gametocyte clearance, because of the risk of life-threatening haemolytic anaemia that can occur in G6PD deficient patients. Although a range of methods for screening G6PD deficiency have been described, almost all require skilled personnel, expensive laboratory equipment, freshly collected blood, and are time consuming; factors that render them unsuitable for mass-screening purposes. Methods A published WST8/1-methoxy PMS method was adapted to assay G6PD activity in a 96-well format using dried blood spots, and used it to undertake population screening within a malaria survey undertaken in Isabel Province, Solomon Islands. The assay results were compared to a biochemical test and a recently marketed rapid diagnostic test. Results Comparative testing with biochemical and rapid diagnostic test indicated that results obtained by filter paper assay were accurate providing that blood spots were assayed within 5 days when stored at ambient temperature and 10 days when stored at 4 degrees. Screening of 8541 people from 41 villages in Isabel Province, Solomon Islands revealed the prevalence of G6PD deficiency as defined by enzyme activity Conclusions The assay enabled simple and quick semi-quantitative population screening in a malaria-endemic region. The study indicated a high prevalence of G6PD deficiency in Isabel Province and highlights the critical need to consider G6PD deficiency in the context of P. vivax malaria elimination strategies in Solomon Islands, particularly in light of the potential role of primaquine mass drug administration.

Radioreceptor assays: plasma membrane receptors and assays for polypeptide and glycoprotein hormones

International Nuclear Information System (INIS)

Schulster, D.

1977-01-01

Receptors for peptide, protein and glycoprotein hormones, and the catecholamines are located on the plasma membranes of their target cells. Preparations of the receptors may be used as specific, high-affinity binding agents for these hormones in assay methodology akin to that for radioimmunoassay. A particular advantage of the radioreceptor assay is that it has a specificity directed towards the biologically active region of the hormone, rather than to some immunologically active region that may have little (or no) involvement in the expression of hormonal activity. Methods for hormone receptor preparation vary greatly, and range from the use of intact cells (as the source of hormone receptor) to the use of purified or solubilized membrane receptors. Receptors isolated from plasma membranes have proved to be of variable stability, and may be damaged during preparation and/or storage. Moreover, since they are present in relatively low concentration in the cell, their preparation in sufficient quantity for use in a radioreceptor assay may present technical problems. In general, there is good correlation between radioreceptor assays and in-vitro bioassays; differences between results from radioreceptor assays and radioimmunoassays are similar to those noted between in-vitro bioassays and radioimmunoassays. The sensitivity of the method is such that normal plasma concentrations of various hormones have been assayed by this technique. (author)

Relationship between the radioisotopic footpad assay and other immunological assays in tumor bearing rats

International Nuclear Information System (INIS)

Mizushima, Yutaka; Takeichi, Noritoshi; Minami, Akio; Kasai, Masaharu; Itaya, Toshiyuki

1981-01-01

KMT-17, a fibrosarcoma induced by 3-methylcholanthrene in a WKA rat, is a sensitive tumor to various kinds of immunological assays and is a suitable model tumor for the study of the immune status in tumor bearing hosts. The antitumor immune response of KMT-17 bearing rats was studied by a radioisotopic footpad assay (FPA) in comparison with other in vivo and in vitro assays. Delayed hypersensitivity to tumor antigens measured by the FPA was observed from the 8th day after transplantation of KMT-17 cells, reached a peak on the 12 - 15th day, and then declined in the late stage on the 17th day. The kinetics of the FPA correlated well with those of an in vivo Winn assay and of an in vitro lymphocyte cytotoxicity assay ( 51 Cr-release assay). The appearance of an antitumor antibody detected by a complement dependent cytotoxicity test also correlated well with the kinetics of the FPA. A growth inhibition assay (GIA) for non-specific cell-mediated immunity also showed similar kinetics to that of the FPA. The delayed hypersensitivity footpad reaction to tumor cell extracts measured by this FPA was tumor-specific. These results suggest that the FPA is a simple and reliable in vivo assay for evaluating antitumor immunity in tumor bearing hosts. (author)

Performance of a Multiplex Serological Helicobacter pylori Assay on a Novel Microfluidic Assay Platform

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Angela Filomena

2017-10-01

Full Text Available Infection with Helicobacter pylori (H. pylori occurs in 50% of the world population, and is associated with the development of ulcer and gastric cancer. Serological diagnostic tests indicate an H. pylori infection by detecting antibodies directed against H. pylori proteins. In addition to line blots, multiplex assay platforms provide smart solutions for the simultaneous analysis of antibody responses towards several H. pylori proteins. We used seven H. pylori proteins (FliD, gGT, GroEL, HpaA, CagA, VacA, and HP0231 and an H. pylori lysate for the development of a multiplex serological assay on a novel microfluidic platform. The reaction limited binding regime in the microfluidic channels allows for a short incubation time of 35 min. The developed assay showed very high sensitivity (99% and specificity (100%. Besides sensitivity and specificity, the technical validation (intra-assay CV = 3.7 ± 1.2% and inter-assay CV = 5.5 ± 1.2% demonstrates that our assay is also a robust tool for the analysis of the H. pylori-specific antibody response. The integration of the virulence factors CagA and VacA allow for the assessment of the risk for gastric cancer development. The short assay time and the performance of the platform shows the potential for implementation of such assays in a clinical setting.

Silver-embedded modified hyperbranched epoxy/clay nanocomposites as antibacterial materials.

Science.gov (United States)

Roy, Buddhadeb; Bharali, Pranjal; Konwar, B K; Karak, Niranjan

2013-01-01

Silver-embedded modified hyperbranched epoxy/clay nanocomposites were prepared at different wt.% of octadecyl amine-modified montmorillonite at a constant silver concentration (1 wt.%). UV-visible, XRD and TEM studies confirmed the formation of silver nanoparticles. Compared to the system without silver and clay, the gloss from 70° to 94°, scratch hardness from 4 to 5.8 kg, impact strength from 60 to 90 cm, tensile strength from 8.5 to 15.5 MPa, adhesive strength from 5 to 7.1 × 10(9)N/m, flexibility from >6 to nanocomposites showed antibacterial activity in well diffusion assays against Staphylococcus aureus (ATCC11632), Bacillus subtilis (ATCC11774), Escherichia coli (MTCC40), Pseudomonas aeruginosa (MTCC7814) and Klebsiella pneumoniae (ATCC10031). The results showed that these nanocomposites have potential to be used as antimicrobial materials. Copyright © 2012 Elsevier Ltd. All rights reserved.

Detection of HLA Antibodies in Organ Transplant Recipients – Triumphs and Challenges of the Solid Phase Bead Assay

Science.gov (United States)

Tait, Brian D.

2016-01-01

This review outlines the development of human leukocyte antigen (HLA) antibody detection assays and their use in organ transplantation in both antibody screening and crossmatching. The development of sensitive solid phase assays such as the enzyme-linked immunosorbent assay technique, and in particular the bead-based technology has revolutionized this field over the last 10–15 years. This revolution however has created a new paradigm in clinical decision making with respect to the detection of low level pretransplant HLA sensitization and its clinical relevance. The relative sensitivities of the assays used are discussed and the relevance of conflicting inter-assay results. Each assay has its advantages and disadvantages and these are discussed. Over the last decade, the bead-based assay utilizing the Luminex® fluorocytometer instrument has become established as the “gold standard” for HLA antibody testing. However, there are still unresolved issues surrounding this technique, such as the presence of denatured HLA molecules on the beads which reveal cryptic epitopes and the issue of appropriate fluorescence cut off values for positivity. The assay has been modified to detect complement binding (CB) in addition to non-complement binding (NCB) HLA antibodies although the clinical relevance of the CB and NCB IgG isotypes is not fully resolved. The increase sensitivity of the Luminex® bead assay over the complement-dependent cytotoxicity crossmatch has permitted the concept of the “virtual crossmatch” whereby the crossmatch is predicted to a high degree of accuracy based on the HLA antibody specificities detected by the solid phase assay. Dialog between clinicians and laboratory staff on an individual patient basis is essential for correct clinical decision making based on HLA antibody results obtained by the various techniques. PMID:28018342

Liquor oligoclonal bands assay: interpretation, correlation with other laboratory assays and importance for diagnostics of neurological disorders

OpenAIRE

Bagdonas, Dovydas

2017-01-01

Aim: to analyse the possible relationship between liquor IgG oligoclonal bands assay and other laboratory assays in neurological patients. Objectives: to determine the frequency of oligoclonal bands in neurological patients; to compare the results between serum and liquor laboratory assays in dependence of oligoclonal bands assay results; to evaluate the relationships between oligoclonal bands assay and serological-immunological assays for infectious diseases, gender, age and neurological ...

A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles

International Nuclear Information System (INIS)

Li, Yaqi; Sun, Li; Qian, Jing; Wang, Chengke; Liu, Qian; Han, En; Hao, Nan; Zhang, Liuping; Cai, Jianrong; Wang, Kun

2016-01-01

In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1–500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection. - Highlights: • Both NGQDs and AgNPs were selected as the novel FRET donor-acceptor pairs. • The proposed homogeneous FRET assay was developed for CaMV35S detection. • The resulting method could identify 0.5% containing transgenic soybean sample. • This assay was inexpensive, simple and highly sensitive.

A homogeneous assay for highly sensitive detection of CaMV35S promoter in transgenic soybean by förster resonance energy transfer between nitrogen-doped graphene quantum dots and Ag nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Li, Yaqi; Sun, Li [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Qian, Jing [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China); Wang, Chengke [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Liu, Qian [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China); Han, En [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Hao, Nan [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China); Zhang, Liuping [Sinograin Zhenjiang Grains & Oils Quality Testing Center Co., Ltd., Zhenjiang, 212013 (China); Cai, Jianrong, E-mail: jrcai@ujs.edu.cn [School of Food and Biological Engineering, Jiangsu University, Zhenjiang, 212013 (China); Wang, Kun, E-mail: wangkun@ujs.edu.cn [School of Chemistry and Chemical Engineering, Jiangsu University, Zhenjiang, 212013 (China)

2016-12-15

In this work, a novel homogeneous assay for DNA quantitative analysis based on förster resonance energy transfer (FRET) was developed for cauliflwer mosaic virus 35s (CaMV35S) promoter of transgenic soybean detection. The homogenous FRET of fluorescence signal was fabricated by DNA hybridization with probe modified nitrogen-doped graphene quantum dots (NGQDs) and silver nanoparticles (AgNPs), which acted the donor-acceptor pairs for the first time. The highly efficient FRET and unique properties of the NGQDs made the proposed FRET system as a functionalized detection platform for labelling of DNA. Upon the recognition of specific target DNA (tDNA), the FRET between NGQDs and AgNPs was triggered to produce fluorescence quenching, which could be used for tDNA detection. The fabricated homogeneous FRET assay displayed a wide linear range of 0.1–500.0 nM and a low limit of detection 0.03 nM for the detection of CaMV35S (S/N = 3). This proposed biosensor revealed high specificity to detect tDNA, with acceptable intra-assay precision and excellent stability. This method was successfully applied to identify the real sample of 0.5% containing transgenic soybean, which achieved the most of national law regulations. This assay was further validated by polymerase chain reaction as the genetically modified organisms, suggesting that the proposed FRET system is a feasible tool for the further daily genetically modified organism detection. - Highlights: • Both NGQDs and AgNPs were selected as the novel FRET donor-acceptor pairs. • The proposed homogeneous FRET assay was developed for CaMV35S detection. • The resulting method could identify 0.5% containing transgenic soybean sample. • This assay was inexpensive, simple and highly sensitive.

Endogenous Locus Reporter Assays.

Science.gov (United States)

Liu, Yaping; Hermes, Jeffrey; Li, Jing; Tudor, Matthew

2018-01-01

Reporter gene assays are widely used in high-throughput screening (HTS) to identify compounds that modulate gene expression. Traditionally a reporter gene assay is built by cloning an endogenous promoter sequence or synthetic response elements in the regulatory region of a reporter gene to monitor transcriptional activity of a specific biological process (exogenous reporter assay). In contrast, an endogenous locus reporter has a reporter gene inserted in the endogenous gene locus that allows the reporter gene to be expressed under the control of the same regulatory elements as the endogenous gene, thus more accurately reflecting the changes seen in the regulation of the actual gene. In this chapter, we introduce some of the considerations behind building a reporter gene assay for high-throughput compound screening and describe the methods we have utilized to establish 1536-well format endogenous locus reporter and exogenous reporter assays for the screening of compounds that modulate Myc pathway activity.

Capillary electrophoresis with electrochemiluminescent detection for highly sensitive assay of genetically modified organisms.

Science.gov (United States)

Guo, Longhua; Yang, Huanghao; Qiu, Bin; Xiao, Xueyang; Xue, Linlin; Kim, Donghwan; Chen, Guonan

2009-12-01

A capillary electrophoresis coupled with electrochemiluminescent detection system (CE-ECL) was developed for the detection of polymerase chain reaction (PCR) amplicons. The ECL luminophore, tris(1,10-phenanthroline) ruthenium(II) (Ru(phen)(3)(2+)), was labeled to the PCR primers before amplification. Ru(phen)(3)(2+) was then introduced to PCR amplicons by PCR amplification. Eventually, the PCR amplicons were separated and detected by the homemade CE-ECL system. The detection of a typical genetically modified organism (GMO), Roundup Ready Soy (RRS), was shown as an example to demonstrate the reliability of the proposed approach. Four pairs of primers were amplified by multiple PCR (MPCR) simultaneously, three of which were targeted on the specific sequence of exogenous genes of RRS, and another was targeted on the endogenous reference gene of soybean. Both the conditions for PCR amplification and CE-ECL separation and detection were investigated in detail. Results showed that, under the optimal conditions, the proposed method can accurately identifying RRS. The corresponding limit of detection (LOD) was below 0.01% with 35 PCR cycles.

Chronocoulometry of wine on multi-walled carbon nanotube modified electrode: Antioxidant capacity assay.

Science.gov (United States)

Ziyatdinova, Guzel; Kozlova, Ekaterina; Budnikov, Herman

2016-04-01

Phenolic antioxidants of wine were electrochemically oxidized on multi-walled carbon nanotubes modified glassy carbon electrode (MWNT/GCE) in phosphate buffer solution. Three oxidation peaks were observed at 0.39, 0.61 and 0.83V for red dry wine and 0.39, 0.80 and 1.18 V for white dry wine, respectively, using differential pulse voltammetry at pH 4.0. The oxidation potentials for individual phenolic antioxidants confirmed the integral nature of the analytical signals for the wines examined. A one-step chronocoulometric method at 0.83 and 1.18 V for red and white wines, respectively, has been developed for the evaluation of wine antioxidant capacity (AOC). The AOC is expressed in gallic acid equivalents per 1L of wine. The AOC of white wine was significantly less than red wine (386 ± 112 vs. 1224 ± 184, pwine and total antioxidant capacity, based on coulometric titration with electrogenerated bromine (r=0.8957 at n=5 and r=0.8986 at n=4 for red and white wines, respectively). Copyright © 2015 Elsevier Ltd. All rights reserved.

Biocompatibility studies of polyacrylonitrile membranes modified with carboxylated polyetherimide

Energy Technology Data Exchange (ETDEWEB)

Senthilkumar, S.; Rajesh, S.; Jayalakshmi, A.; Mohan, D., E-mail: mohantarun@gmail.com

2013-10-15

Poly (ether-imide) (PEI) was carboxylated and used as the hydrophilic modification agent for the preparation of polyacrylonitrile (PAN) membranes. Membranes were prepared with different blend compositions of PAN and CPEI by diffusion induced precipitation. The modified membranes were characterized by thermo gravimetric analysis (TGA), mechanical analysis, scanning electron microscopy (SEM) and contact angle measurement to understand the influence of CPEI on the properties of the membranes. The biocompatibility studies exhibited reduced plasma protein adsorption, platelet adhesion and thrombus formation on the modified membrane surface. The complete blood count (CBC) results of CPEI incorporated membranes showed stable CBC values and significant decrease in the complement activation were also observed. In addition to good cytocompatibility, monocytes cultured on these modified membranes exhibited improved functional profiles in 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Thus it could be concluded that PAN/CPEI membranes with excellent biocompatibility can be useful for hemodialysis. Highlights: • Carboxylated PEI was prepared and utilized as hydrophilic modification agent. • CPEI incorporated into PAN to improved biocompatibility and cyto compatibility • Biocompatibility of membranes was correlated with morphology and hydrophilicity. • Antifouling studies of the PAN/CPEI membranes was studied by BSA as model foulant.

Activity-based assay for human mono-ADP-ribosyltransferases ARTD7/PARP15 and ARTD10/PARP10 aimed at screening and profiling inhibitors.

Science.gov (United States)

Venkannagari, Harikanth; Fallarero, Adyary; Feijs, Karla L H; Lüscher, Bernhard; Lehtiö, Lari

2013-05-13

Poly(ADP-ribose) polymerases (PARPs) or diphtheria toxin like ADP-ribosyl transferases (ARTDs) are enzymes that catalyze the covalent modification of proteins by attachment of ADP-ribose units to the target amino acid residues or to the growing chain of ADP-ribose. A subclass of the ARTD superfamily consists of mono-ADP-ribosyl transferases that are thought to modify themselves and other substrate proteins by covalently adding only a single ADP-ribose moiety to the target. Many of the ARTD enzymes are either established or potential drug targets and a functional activity assay for them will be a valuable tool to identify selective inhibitors for each enzyme. Existing assays are not directly applicable for screening of inhibitors due to the different nature of the reaction and different target molecules. We modified and applied a fluorescence-based assay previously described for PARP1/ARTD1 and tankyrase/ARTD5 for screening of PARP10/ARTD10 and PARP15/ARTD7 inhibitors. The assay measures the amount of NAD(+) present after chemically converting it to a fluorescent analog. We demonstrate that by using an excess of a recombinant acceptor protein the performance of the activity-based assay is excellent for screening of compound libraries. The assay is homogenous and cost effective, making it possible to test relatively large compound libraries. This method can be used to screen inhibitors of mono-ARTDs and profile inhibitors of the enzyme class. The assay was optimized for ARTD10 and ARTD7, but it can be directly applied to other mono-ARTDs of the ARTD superfamily. Profiling of known ARTD inhibitors against ARTD10 and ARTD7 in a validatory screening identified the best inhibitors with submicromolar potencies. Only few of the tested ARTD inhibitors were potent, implicating that there is a need to screen new compound scaffolds. This is needed to create small molecules that could serve as biological probes and potential starting points for drug discovery projects against

Optimization of digital droplet polymerase chain reaction for quantification of genetically modified organisms.

Science.gov (United States)

Gerdes, Lars; Iwobi, Azuka; Busch, Ulrich; Pecoraro, Sven

2016-03-01

Digital PCR in droplets (ddPCR) is an emerging method for more and more applications in DNA (and RNA) analysis. Special requirements when establishing ddPCR for analysis of genetically modified organisms (GMO) in a laboratory include the choice between validated official qPCR methods and the optimization of these assays for a ddPCR format. Differentiation between droplets with positive reaction and negative droplets, that is setting of an appropriate threshold, can be crucial for a correct measurement. This holds true in particular when independent transgene and plant-specific reference gene copy numbers have to be combined to determine the content of GM material in a sample. Droplets which show fluorescent units ranging between those of explicit positive and negative droplets are called 'rain'. Signals of such droplets can hinder analysis and the correct setting of a threshold. In this manuscript, a computer-based algorithm has been carefully designed to evaluate assay performance and facilitate objective criteria for assay optimization. Optimized assays in return minimize the impact of rain on ddPCR analysis. We developed an Excel based 'experience matrix' that reflects the assay parameters of GMO ddPCR tests performed in our laboratory. Parameters considered include singleplex/duplex ddPCR, assay volume, thermal cycler, probe manufacturer, oligonucleotide concentration, annealing/elongation temperature, and a droplet separation evaluation. We additionally propose an objective droplet separation value which is based on both absolute fluorescence signal distance of positive and negative droplet populations and the variation within these droplet populations. The proposed performance classification in the experience matrix can be used for a rating of different assays for the same GMO target, thus enabling employment of the best suited assay parameters. Main optimization parameters include annealing/extension temperature and oligonucleotide concentrations. The

ASSESSING POSSIBLE ECOLOGICAL RISKS OF GENETICALLY MODIFIED CROPS: GENE EXPRESSION ASSAYS AND GENETIC MONITORING OF NON-TARGET ORGANISMS

Science.gov (United States)

Widespread planting of genetically modified crops with the Bt transgene pesticide has led to concern over non-target effects of Bt compounds in agroecosystems. While some research suggests that non-target organisms exposed to Bt toxin exhibit reduced fecundity and increased morta...

CYP1A inhibition in fish gill filaments: A novel assay applied on pharmaceuticals and other chemicals

Energy Technology Data Exchange (ETDEWEB)

Beijer, Kristina; Abrahamson, Alexandra; Brunstroem, Bjoern [Department of Environmental Toxicology, Uppsala University, Norbyvaegen 18A, SE-752 36 Uppsala (Sweden); Brandt, Ingvar, E-mail: ingvar.brandt@ebc.uu.se [Department of Environmental Toxicology, Uppsala University, Norbyvaegen 18A, SE-752 36 Uppsala (Sweden)

2010-01-31

The gill filament 7-ethoxyresorufin O-deethylase (EROD) assay was originally developed as a biomarker for cytochrome P4501A (CYP1A) induction by Ah-receptor agonists in water. In this study, the assay was adapted to measure inhibition of CYP1A activity in fish gill filaments ex vivo. The experiments were carried out using gill arch filaments from {beta}-naphthoflavone ({beta}NF)-exposed three-spined stickleback (Gasterosteus aculeatus). Candidate CYP1A inhibitors were added to the assay buffer. Nine selected pharmaceuticals and five known or suspected CYP1A-modulating chemicals were examined with regard to their ability to reduce EROD activity in gill filaments. Ellipticine, a well characterized CYP1A inhibitor, was the most effective inhibitor of the compounds tested. At a concentration in the assay buffer of 1 {mu}M the antifungal azoles ketoconazole, miconazole and bitertanol, and the plant flavonoid acacetin reduced gill EROD activity by more than 50%, implying IC50 values below 1 {mu}M. These compounds have previously been shown to inhibit EROD activity in liver microsomes from fish and mammals at similar concentrations. The proton pump inhibitor omeprazole reduced the gill EROD activity by 39% at 10 {mu}M. It is concluded that the modified gill filament EROD assay is useful to screen for waterborne pollutants that inhibit catalytic CYP1A activity in fish gills.

CYP1A inhibition in fish gill filaments: A novel assay applied on pharmaceuticals and other chemicals

International Nuclear Information System (INIS)

Beijer, Kristina; Abrahamson, Alexandra; Brunstroem, Bjoern; Brandt, Ingvar

2010-01-01

The gill filament 7-ethoxyresorufin O-deethylase (EROD) assay was originally developed as a biomarker for cytochrome P4501A (CYP1A) induction by Ah-receptor agonists in water. In this study, the assay was adapted to measure inhibition of CYP1A activity in fish gill filaments ex vivo. The experiments were carried out using gill arch filaments from β-naphthoflavone (βNF)-exposed three-spined stickleback (Gasterosteus aculeatus). Candidate CYP1A inhibitors were added to the assay buffer. Nine selected pharmaceuticals and five known or suspected CYP1A-modulating chemicals were examined with regard to their ability to reduce EROD activity in gill filaments. Ellipticine, a well characterized CYP1A inhibitor, was the most effective inhibitor of the compounds tested. At a concentration in the assay buffer of 1 μM the antifungal azoles ketoconazole, miconazole and bitertanol, and the plant flavonoid acacetin reduced gill EROD activity by more than 50%, implying IC50 values below 1 μM. These compounds have previously been shown to inhibit EROD activity in liver microsomes from fish and mammals at similar concentrations. The proton pump inhibitor omeprazole reduced the gill EROD activity by 39% at 10 μM. It is concluded that the modified gill filament EROD assay is useful to screen for waterborne pollutants that inhibit catalytic CYP1A activity in fish gills.

Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

International Nuclear Information System (INIS)

Pastrana, Diana V.; Buck, Christopher B.; Pang, Y.-Y. S.; Thompson, Cynthia D.; Castle, Philip E.; FitzGerald, Peter C.; Krueger Kjaer, Susanne; Lowy, Douglas R.; Schiller, John T.

2004-01-01

Sensitive high-throughput neutralization assays, based upon pseudoviruses carrying a secreted alkaline phosphatase (SEAP) reporter gene, were developed and validated for human papillomavirus (HPV)16, HPV18, and bovine papillomavirus 1 (BPV1). SEAP pseudoviruses were produced by transient transfection of codon-modified papillomavirus structural genes into an SV40 T antigen expressing line derived from 293 cells, yielding sufficient pseudovirus from one flask for thousands of titrations. In a 96-well plate format, in this initial characterization, the assay was reproducible and appears to be as sensitive as, but more specific than, a standard papillomavirus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). The neutralization assay detected type-specific HPV16 or HPV18 neutralizing antibodies (titers of 160-10240) in sera of the majority of a group of women infected with the corresponding HPV type, but not in virgin women. Sera from HPV16 VLP vaccinees had high anti-HPV16 neutralizing titers (mean: 45000; range: 5120-163840), but no anti-HPV18 neutralizing activity. The SEAP pseudovirus-based neutralization assay should be a practical method for quantifying potentially protective antibody responses in HPV natural history and prophylactic vaccine studies

Monitoring of DNA breakage in embryonic stages of the African catfish Clarias gariepinus (Burchell, 1822) after exposure to lead nitrate using alkaline comet assay

NARCIS (Netherlands)

Osman, A.G.M.; Mekkawy, Imam A.; Verreth, J.A.J.; Wuertz, S.; Kloas, W.; Kirschbaum, Frank

2008-01-01

Increasing lead contamination in Egyptian ecosystems and high lead concentrations in food items have raised concern for human health and stimulated studies on monitoring ecotoxicological impact of lead-caused genotoxicity. In this work, the alkaline comet assay was modified for monitoring DNA strand

Benefits of gene transduction of granulocyte macrophage colony-stimulating factor in cancer vaccine using genetically modified dendritic cells.

Science.gov (United States)

Ojima, Toshiyasu; Iwahashi, Makoto; Nakamura, Masaki; Matsuda, Kenji; Nakamori, Mikihito; Ueda, Kentaro; Naka, Teiji; Katsuda, Masahiro; Miyazawa, Motoki; Yamaue, Hiroki

2007-10-01

Granulocyte macrophage colony-stimulating factor (GM-CSF) is a key cytokine for the generation and stimulation of dendritic cells (DCs), and it may also play a pivotal role in promoting the survival of DCs. In this study, the feasibility of creating a cancer vaccine using DCs adenovirally transduced with the carcinoembryonic antigen (CEA) gene and the GM-CSF gene was examined. In addition, the effect of the co-transduction of GM-CSF gene on the lifespan of these genetically modified DCs was determined. A cytotoxic assay using peripheral blood mononuclear cell (PBMC)-derived cytotoxic T lymphocytes (CTLs) was performed in a 4-h 51Cr release assay. The apoptosis of DCs was examined by TdT-mediated dUTP-FITC nick end labeling (TUNEL) assay. CEA-specific CTLs were generated from PBMCs stimulated with genetically modified DCs expressing CEA. The cytotoxicity of these CTLs was augmented by co-transduction of DCs with the GM-CSF gene. Co-transduction of the GM-CSF gene into DCs inhibited apoptosis of these DCs themselves via up-regulation of Bcl-x(L) expression, leading to the extension of the lifespan of these DCs. Furthermore, the transduction of the GM-CSF gene into DCs also suppressed the incidence of apoptosis of DCs induced by transforming growth factor-beta1 (TGFbeta-1). Immunotherapy using these genetically modified DCs may therefore be useful with several advantages as follows: i) adenoviral toxicity to DCs can be reduced; ii) the lifespan of vaccinated DCs can be prolonged; and iii) GM-CSF may protect DCs from apoptosis induced by tumor-derived TGFbeta-1 in the regional lymph nodes.

Direct 125I-radioligand assays for serum progesterone compared with assays involving extraction of serum

International Nuclear Information System (INIS)

Ratcliffe, W.A.; Corrie, J.E.T.; Dalziel, A.H.; Macpherson, J.S.

1982-01-01

Two direct radioimmunoassays for progesterone in 50 μL of unextracted serum or plasma with assays involving extraction of serum were compared. The direct assays include the use of either danazol at pH 7.4 or 8-anilino-1-naphthalenesulfonic acid at pH 4.0 to displace progesterone from serum binding-proteins. Progesterone is then assayed by using an antiserum to a progesterone 11α-hemisuccinyl conjugate and the radioligand 125 I-labeled progesterone 11α-glucuronyl tyramine, with separation by double-antibody techniques. Direct assays with either displacing agent gave good analytical recovery of progesterone added to human serum, and progesterone values for patients' specimens correlated well (r > 0.96) with results of assays involving extraction of serum. Precision was similar with each displacing agent over the working range 2.5-100 nmol/L and superior to that of extraction assays. We conclude that these direct assays of progesterone are analytically valid and more robust, precise, and technically convenient than many conventional methods involving extraction of serum

Absolute nuclear material assay

Science.gov (United States)

Prasad, Manoj K [Pleasanton, CA; Snyderman, Neal J [Berkeley, CA; Rowland, Mark S [Alamo, CA

2010-07-13

A method of absolute nuclear material assay of an unknown source comprising counting neutrons from the unknown source and providing an absolute nuclear material assay utilizing a model to optimally compare to the measured count distributions. In one embodiment, the step of providing an absolute nuclear material assay comprises utilizing a random sampling of analytically computed fission chain distributions to generate a continuous time-evolving sequence of event-counts by spreading the fission chain distribution in time.

A practicable detection system for genetically modified rice by SERS-barcoded nanosensors.

Science.gov (United States)

Chen, Kun; Han, Heyou; Luo, Zhihui; Wang, Yanjun; Wang, Xiuping

2012-04-15

Since the global cultivation of genetically modified crops constantly expands, it remains a high demand to establish different ways to sort food and feed that consist or contain genetically modified organisms. Surface-enhanced Raman scattering (SERS) spectroscopy is a flexible tool for biological analysis due to its excellent properties for detecting wide varieties of target biomolecules including nucleic acids. In the present study, a SERS-barcoded nanosensor was developed to detect Bacillus thuringiensis (Bt) gene-transformed rice expressing insecticidal proteins. The barcoded sensor was designed by encapsulation of gold nanoparticles with silica and conjugation of oligonucleotide strands for targeting DNA strands. The transition between the cry1A(b) and cry1A(c) fusion gene sequence was used to construct a specific SERS-based detection method with a detection limit of 0.1 pg/mL. In order to build the determination models to screen transgene, a series mixture of Bt rice and normal rice were prepared for SERS assay, and the limit of detection was 0.1% (w/w) transgenic Bt rice relative to normal rice. The sensitivity and accuracy of the SERS-based assay was comparable with real-time PCR. The SERS-barcoded analytical method would provide precise detection of transgenic rice varieties but also informative supplement to avoid false positive outcomes. Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.

Modified beacon probe assisted dual signal amplification for visual detection of microRNA.

Science.gov (United States)

Sun, Xiuwei; Ying, Na; Ju, Chuanjing; Li, Zhongyi; Xu, Na; Qu, Guijuan; Liu, Wensen; Wan, Jiayu

2018-04-21

In a recent study, we reported a novel assay for the detection of microRNA-21 based on duplex-specific nuclease (DSN)-assisted isothermal cleavage and hybridization chain reaction (HCR) dual signal amplification. The Fam modified double-stranded DNA products were generated after the HCR, another biotin modified probe was digested by DSN and released from the magnetic beads after the addition of the target miRNA. The released sequence was then combined with HCR products to form a double-tagging dsDNA, which can be recognized by the lateral flow strips. In this study, we introduced a 2-OMethyl-RNA modified beacon probe (2-OMe-MB) to make some improvements based on the previous study. Firstly, the substitution of modified probe combined on magnetic beads avoids the fussy washing steps for the separation of un-reacted probes. Furthermore, the modification of 2-OMe on the stem of the probe avoided the unnecessary cleavage by DSN, which greatly reduce the background signal. Compared to the previous work, these improvements save us a lot of steps but possess the comparable sensitivity and selectivity. Copyright © 2018 Elsevier Inc. All rights reserved.

Evaluation of the highly sensitive Roche thyroglobulin II assay and establishment of a reference limit for thyroglobulin-negative patient samples.

Science.gov (United States)

Rotteveel-de Groot, Dorien M; Ross, H Alec; Janssen, Marcel J R; Netea-Maier, Romana T; Oosting, Janine D; Sweep, Fred C G J; van Herwaarden, Antonius E

2016-08-01

Thyroglobulin (Tg) measurements are used to monitor for residual thyroid tissue in patients with differentiated thyroid cancer (DTC) after thyroidectomy and radioiodine ablative therapy. In recent years highly sensitive Tg assays have been developed. In this study the analytical performance of the new Roche Elecsys Tg II assay was evaluated and compared with the well documented Access2 Tg assay (Beckman-Coulter). Analytical performance was examined using various Clinical and Laboratory Standards Institute (CLSI) evaluation protocols. Tg negative patient sera were used to establish an upper reference limit (URL) for the Elecsys Tg II assay. Non-linearity, drift and carry-over according to CLSI EP10 and EP6 in a measuring range of 0.04-500 ng/mL were non-significant. Total precision according to CLSI EP5 was 10% at a Tg concentration of 0.08 ng/mL. A patient serum comparison performed according to a modified CLSI EP9 protocol showed a significant difference of a factor of approximately 1.4, despite using an identical CRM calibrator. The Elecsys Tg II assay measured Tg with a two-fold higher sensitivity than the Access2 assay. Finally, using human sera without Tg, an URL of 0.05 ng/mL was determined. In our hands the highly sensitive Elecsys Tg II assay shows a good analytical performance and a higher sensitivity compared to the Access2 Tg assay. An URL of 0.05 ng/mL for the Elecsys Tg II assay was determined which may improve the clinical utility of the assay for the detection of residual DTC or disease recurrence.

Ultrasensitive electrochemiluminescent immunoassay for morphine using a gold electrode modified with CdS quantum dots, polyamidoamine, and gold nanoparticles

International Nuclear Information System (INIS)

Fei, Wenjuan; Chen, Feifei; Sun, Li; Li, Qianhua; Wu, Ying; Yang, Jianping

2014-01-01

We report on a novel electrochemiluminescent (ECL) immunoassay for the ultrasensitive determination of morphine by making use of a gold electrode which was modified with a nanocomposite film containing self-assembled polyamidoamine (PAMAM) CdS quantum dots and electrodeposited gold nanoparticles (Au-NPs). The highly uniform and well-dispersed quantum dots were capped with PAMAM dendrimers. Due to the synergistic effect of the modified quantum dots and the electrodeposited Au-NPs, the ECL response is dramatically enhanced. Under optimal experimental conditions, the immunoreaction between morphine and anti-morphine antibody resulted in a decrease of the ECL signal because of steric hindrance. The calibration plot is linear in the morphine concentration range from 0.2 to 180 ng•mL −1 , with a detection limit as low as 67 pg•mL −1 . The sensor was successfully applied to the determination of morphine in blood plasma. This kind of assay is expected to pave new avenues in label-free drug assays. (author)

Detection of Dirofilaria immitis and other arthropod-borne filarioids by an HRM real-time qPCR, blood-concentrating techniques and a serological assay in dogs from Costa Rica.

Science.gov (United States)

Rojas, Alicia; Rojas, Diana; Montenegro, Víctor M; Baneth, Gad

2015-03-23

Canine filarioids are important nematodes transmitted to dogs by arthropods. Diagnosis of canine filariosis is accomplished by the microscopic identification of microfilariae, serology or PCR for filarial-DNA. The aim of this study was to evaluate a molecular assay for the detection of canine filariae in dog blood, to compare its performance to other diagnostic techniques, and to determine the relationship between microfilarial concentration and infection with other vector-borne pathogens. Blood samples from 146 dogs from Costa Rica were subjected to the detection of canine filarioids by four different methods: the microhematocrit tube test (MCT), Knott's modified test, serology and a high resolution melt and quantitative real-time PCR (HRM-qPCR). Co-infection with other vector-borne pathogens was also evaluated. Fifteen percent of the dogs were positive to Dirofilaria immitis by at least one of the methods. The HRM-qPCR produced distinctive melting plots for the different filarial worms and revealed that 11.6% of dogs were infected with Acanthocheilonema reconditum. The latter assay had a limit of detection of 2.4x10⁻⁴ mf/μl and detected infections with lower microfilarial concentrations in comparison to the microscopic techniques and the serological assay. The MCT and Knott's test only detected dogs with D. immitis microfilaremias above 0.7 mf/μl. Nevertheless, there was a strong correlation between the microfilarial concentration obtained by the Knott's modified test and the HRM-qPCR (r = 0.906, p HRM-qPCR showed the most sensitive and reliable performance in the detection of blood filaroids in comparison to the Knott's modified test, the MCT test and a serological assay.

Evaluation of the highly sensitive Roche thyroglobulin II assay and establishment of a reference limit for thyroglobulin-negative patient samples

Directory of Open Access Journals (Sweden)

Dorien M. Rotteveel-de Groot

2016-08-01

Full Text Available Objectives: Thyroglobulin (Tg measurements are used to monitor for residual thyroid tissue in patients with differentiated thyroid cancer (DTC after thyroidectomy and radioiodine ablative therapy. In recent years highly sensitive Tg assays have been developed. In this study the analytical performance of the new Roche Elecsys Tg II assay was evaluated and compared with the well documented Access2 Tg assay (Beckman–Coulter. Design and methods: Analytical performance was examined using various Clinical and Laboratory Standards Institute (CLSI evaluation protocols. Tg negative patient sera were used to establish an upper reference limit (URL for the Elecsys Tg II assay. Results: Non-linearity, drift and carry-over according to CLSI EP10 and EP6 in a measuring range of 0.04–500 ng/mL were non-significant. Total precision according to CLSI EP5 was 10% at a Tg concentration of 0.08 ng/mL. A patient serum comparison performed according to a modified CLSI EP9 protocol showed a significant difference of a factor of approximately 1.4, despite using an identical CRM calibrator. The Elecsys Tg II assay measured Tg with a two-fold higher sensitivity than the Access2 assay. Finally, using human sera without Tg, an URL of 0.05 ng/mL was determined. Conclusions: In our hands the highly sensitive Elecsys Tg II assay shows a good analytical performance and a higher sensitivity compared to the Access2 Tg assay. An URL of 0.05 ng/mL for the Elecsys Tg II assay was determined which may improve the clinical utility of the assay for the detection of residual DTC or disease recurrence. Keywords: Thyroglobulin, Roche Elecsys Tg II assay, validation, reporting limit