Accurate mass-based analysis in human sport doping control : Application of liquid chromatography - time-of-flight mass spectrometry

Human sport doping control analysis is a complex and challenging task for anti-doping laboratories. The List of Prohibited Substances and Methods, updated annually by World Anti-Doping Agency (WADA), consists of hundreds of chemically and pharmacologically different low and high molecular weight compounds. This poses a considerable challenge for laboratories to analyze for them all in a limited amount of time from a limited sample aliquot. The continuous expansion of the Prohibited List obliges laboratories to keep their analytical methods updated and to research new available methodologies. In this thesis, an accurate mass-based analysis employing liquid chromatography - time-of-flight mass spectrometry (LC-TOFMS) was developed and validated to improve the power of doping control analysis. New analytical methods were developed utilizing the high mass accuracy and high information content obtained by TOFMS to generate comprehensive and generic screening procedures. The suitability of LC-TOFMS for comprehensive screening was demonstrated for the first time in the field with mass accuracies better than 1 mDa. Further attention was given to generic sample preparation, an essential part of screening analysis, to rationalize the whole work flow and minimize the need for several separate sample preparation methods. Utilizing both positive and negative ionization allowed the detection of almost 200 prohibited substances. Automatic data processing produced a Microsoft Excel based report highlighting the entries fulfilling the criteria of the reverse data base search (retention time (RT), mass accuracy, isotope match). The quantitative performance of LC-TOFMS was demonstrated with morphine, codeine and their intact glucuronide conjugates. After a straightforward sample preparation the compounds were analyzed directly without the need for hydrolysis, solvent transfer, evaporation or reconstitution. The hydrophilic interaction technique (HILIC) provided good chromatographic separation, which was critical for the morphine glucuronide isomers. A wide linear range (50-5000 ng/ml) with good precision (RSD<10%) and accuracy (±10%) was obtained, showing comparable or better performance to other methods used. In-source collision-induced dissociation (ISCID) allowed confirmation analysis with three diagnostic ions with a median mass accuracy of 1.08 mDa and repeatable ion ratios fulfilling WADA s identification criteria. The suitability of LC-TOFMS for screening of high molecular weight doping agents was demonstrated with plasma volume expanders (PVE), namely dextran and hydroxyethylstarch (HES). Specificity of the assay was improved, since interfering matrix compounds were removed by size exclusion chromatography (SEC). ISCID produced three characteristic ions with an excellent mean mass accuracy of 0.82 mDa at physiological concentration levels. In summary, by combining TOFMS with a proper sample preparation and chromatographic separation, the technique can be utilized extensively in doping control laboratories for comprehensive screening of chemically different low and high molecular weight compounds, for quantification of threshold substances and even for confirmation. LC-TOFMS rationalized the work flow in doping control laboratories by simplifying the screening scheme, expediting reporting and minimizing the analysis costs. Therefore LC-TOFMS can be exploited widely in doping control, and the need for several separate analysis techniques is reduced.

Rocking response of rectangular rigid blocks under random noise base excitations

The response of a rigid rectangular block resting on a rigid foundation and acted upon simultaneously by a horizontal and a vertical random white-noise excitation is considered. In the equation of motion, the energy dissipation is modeled through a viscous damping term. Under the assumption that the body does not topple, the steady-state joint probability density function of the rotation and the rotational velocity is obtained using the Fokker-Planck equation approach. Closed form solution is obtained for a specific combination of system parameters. A more general but approximate solution to the joint probability density function based on the method of equivalent non-linearization is also presented. Further, the problem of overturning of the block is approached in the framework of the diffusion methods for first passage failure studies. The overturning of the block is deemed incipient when the response trajectories in the phase plane cross the separatrix of the conservative unforced system. Expressions for the moments of first passage time are obtained via a series solution to the governing generalized Pontriagin-Vitt equations. Numerical results illustra- tive of the theoretical solutions are presented and their validity is examined through limited amount of digital simulations.

Adaptive Flight-Control Design Using Neural-Network-Aided Optimal Nonlinear Dynamic Inversion

A neural-network-aided nonlinear dynamic inversion-based hybrid technique of model reference adaptive control flight-control system design is presented in this paper. Here, the gains of the nonlinear dynamic inversion-based flight-control system are dynamically selected in such a manner that the resulting controller mimics a single network, adaptive control, optimal nonlinear controller for state regulation. Traditional model reference adaptive control methods use a linearized reference model, and the presented control design method employs a nonlinear reference model to compute the nonlinear dynamic inversion gains. This innovation of designing the gain elements after synthesizing the single network adaptive controller maintains the advantages that an optimal controller offers, yet it retains a simple closed-form control expression in state feedback form, which can easily be modified for tracking problems without demanding any a priori knowledge of the reference signals. The strength of the technique is demonstrated by considering the longitudinal motion of a nonlinear aircraft system. An extended single network adaptive control/nonlinear dynamic inversion adaptive control design architecture is also presented, which adapts online to three failure conditions, namely, a thrust failure, an elevator failure, and an inaccuracy in the estimation of C-M alpha. Simulation results demonstrate that the presented adaptive flight controller generates a near-optimal response when compared to a traditional nonlinear dynamic inversion controller.

Multi-scale modelling of Suzuki segregation in gamma' precipitates in Ni and Co-base superalloys

The high temperature strength of alloys with (gamma +gamma') microstructure is primarily due to the resistance of the ordered precipitate to cutting by matrix dislocations. Such shearing requires higher stresses since it involves the creation of a planar fault. Planar fault energy is known to be dependent on composition. This implies that the composition on the fault may be different from that in the bulk for energetic reasons. Such segregation (or desegregation) of specific alloying elements to the fault may result in Suzuki strengthening which has not been explored extensively in these systems. In this work, segregation (or desegregation) of alloying elements to planar faults was studied computationally in Ni-3(Al, Ti) and Co-3(W, Al) type gamma' precipitates. The composition dependence of APB energy and heat of mixing were evaluated from first principle electronic structure calculations. A phase field model incorporating the first principles results, was used to simulate the motion of an extended superdislocation under stress concurrently with composition evolution. Results reveal that in both systems, significant (de) segregation occurs on equilibration. On application of stress, solutes were dragged along with the APB in some cases. Additionally, it was also noted the velocity of the superdislocation under an applied stress is strongly dependent on atomic mobility (i. e. diffusivity).

Structures on sliding base subject to horizontal and vertical motions

In this paper particular investigation is directed towards the combined effects of horizontal and vertical motions of real earthquakes to structures resting on sliding base. A simplified method is presented to treat the nonlinear effects of time dependent frictional force of the sliding base as a function of the vertical reaction produced by the foundation. As an example, the El Centro 1940 earthquake record is used on a structural model to show the structural responses due to a sliding base with different frictional and stiffness characteristics. The study shows that vertical ground motion does affect both the superstructure response and the base sliding displacement. Nevertheless, the sliding base isolator is shown to be effective for the reduction of seismic response of a superstructure.

Octopamine neurons mediate flight-induced modulation of visual processing in Drosophila melanogaster

Activity-dependent modulation of sensory systems has been documented in many organisms, and is likely to be essential for appropriate processing of information during different behavioral states. However, the mechanisms underlying these phenomena, and often their functional consequences, remain poorly characterized. I investigated the role of octopamine neurons in the flight-dependent modulation observed in visual interneurons in the fruit fly Drosophila melanogaster. The vertical system (VS) cells exhibit a boost in their response to visual motion during flight compared to quiescence. Pharmacological application of octopamine evokes responses in quiescent flies that mimic those observed during flight, and octopamine neurons that project to the optic lobes increase in activity during flight. Using genetic tools to manipulate the activity of octopamine neurons, I find that they are both necessary and sufficient for the flight-induced visual boost. This work provides the first evidence that endogenous release of octopamine is involved in state-dependent modulation of visual interneurons in flies. Further, I investigated the role of a single pair of octopamine neurons that project to the optic lobes, and found no evidence that chemical synaptic transmission via these neurons is necessary for the flight boost. However, I found some evidence that activation of these neurons may contribute to the flight boost. Wind stimuli alone are sufficient to generate transient increases in the VS cell response to motion vision, but result in no increase in baseline membrane potential. These results suggest that the flight boost originates not from a central command signal during flight, but from mechanosensory stimuli relayed via the octopamine system. Lastly, in an attempt to understand the functional consequences of the flight boost observed in visual interneurons, we measured the effect of inactivating octopamine neurons in freely flying flies. We found that flies whose octopamine neurons we silenced accelerate less than wild-type flies, consistent with the hypothesis that the flight boost we observe in VS cells is indicative of a gain control mechanism mediated by octopamine neurons. Together, this work serves as the basis for a mechanistic and functional understanding of octopaminergic modulation of vision in flying flies.

Unsteady shock motion on a NACA0012 aerofoil at low reduced frequencies

Using transonic blowdown windtunnel experiments, the 2D unsteady shock motion on a NACA0012 aerofoil is examined at various frequencies typical for helicopter blades in forward flight. The aerofoil is subjected to freestream velocities oscillating periodically between M = 0.66 and M = 0.77. Unsteady pressure traces and schlieren images are analyzed over a range of low reduced frequencies to provide information on shock location and strength throughout the cycle. Unsteady effects were noticeable even at very low reduced frequencies (down to O(0.01). However, through the range of frequencies investigated, and within experimental error, the unsteady shock location showed no discernible lag compared to the quasi-steady behaviour. On the other hand, significant variations were observed in shock strengths with the upstream running part of the cycle (decreasing Mach number) displaying considerably stronger shocks than during the accelerating part of the cycle. It could be shown that this variation in shock strength is primarily caused by the shock motion modifying the relative shock Mach number. As a result is was possible to use the quasi-steady results to predict the unsteady shock behaviour at the frequencies investigated here (below 0(0.1)).

Unsteady shock motion on a NACA0012 aerofoil at low reduced frequencies

Using transonic blowdown windtunnel experiments, the 2D unsteady shock motion on a NACA0012 aerofoil is examined at various frequencies typical for helicopter blades in forward flight. The aerofoil is subjected to freestream velocities oscillating periodically between M = 0.66 and M = 0.77. Unsteady pressure traces and schlieren images are analyzed over a range of low reduced frequencies to provide information on shock location and strength throughout the cycle. Unsteady effects were noticeable even at very low reduced frequencies (down to O(0.01). However, through the range of frequencies investigated, and within experimental error, the unsteady shock location showed no discernible lag compared to the quasi-steady behaviour. On the other hand, significant variations were observed in shock strengths with the upstream running part of the cycle (decreasing Mach number) displaying considerably stronger shocks than during the accelerating part of the cycle. It could be shown that this variation in shock strength is primarily caused by the shock motion modifying the relative shock Mach number. As a result is was possible to use the quasi-steady results to predict the unsteady shock behaviour at the frequencies investigated here (below 0(0.1)).

Swimming direction reversal of flagella through ciliary motion of mastigonemes.

Bio-inspired designs can provide an answer to engineering problems such as swimming strategies at the micron or nano-scale. Scientists are now designing artificial micro-swimmers that can mimic flagella-powered swimming of micro-organisms. In an application such as lab-on-a-chip in which micro-object manipulation in small flow geometries could be achieved by micro-swimmers, control of the swimming direction becomes an important aspect for retrieval and control of the micro-swimmer. A bio-inspired approach for swimming direction reversal (a flagellum bearing mastigonemes) can be used to design such a system and is being explored in the present work. We analyze the system using a computational framework in which the equations of solid mechanics and fluid dynamics are solved simultaneously. The fluid dynamics of Stokes flow is represented by a 2D Stokeslets approach while the solid mechanics behavior is realized using Euler-Bernoulli beam elements. The working principle of a flagellum bearing mastigonemes can be broken up into two parts: (1) the contribution of the base flagellum and (2) the contribution of mastigonemes, which act like cilia. These contributions are counteractive, and the net motion (velocity and direction) is a superposition of the two. In the present work, we also perform a dimensional analysis to understand the underlying physics associated with the system parameters such as the height of the mastigonemes, the number of mastigonemes, the flagellar wave length and amplitude, the flagellum length, and mastigonemes rigidity. Our results provide fundamental physical insight on the swimming of a flagellum with mastigonemes, and it provides guidelines for the design of artificial flagellar systems.

Transducer and base compliance alter the in situ 6 dof force measured from muscle during an isometric contraction in a multi-joint limb.

Although musculoskeletal models are commonly used, validating the muscle actions predicted by such models is often difficult. In situ isometric measurements are a possible solution. The base of the skeleton is immobilized and the endpoint of the limb is rigidly attached to a 6-axis force transducer. Individual muscles are stimulated and the resulting forces and moments recorded. Such analyses generally assume idealized conditions. In this study we have developed an analysis taking into account the compliances due to imperfect fixation of the skeleton, imperfect attachment of the force transducer, and extra degrees of freedom (dof) in the joints that sometimes become necessary in fixed end contractions. We use simulations of the rat hindlimb to illustrate the consequences of such compliances. We show that when the limb is overconstrained, i.e., when there are fewer dof within the limb than are restrained by the skeletal fixation, the compliances of the skeletal fixation and of the transducer attachment can significantly affect measured forces and moments. When the limb dofs and restrained dofs are matched, however, the measured forces and moments are independent of these compliances. We also show that this framework can be used to model limb dofs, so that rather than simply omitting dofs in which a limb does not move (e.g., abduction at the knee), the limited motion of the limb in these dofs can be more realistically modeled as a very low compliance. Finally, we discuss the practical implications of these results to experimental measurements of muscle actions.

A laboratory-scale buried charge simulator

An experimental technique has been developed in order to mimic the effect of landmine loading on materials and structures to be studied in a laboratory setting, without the need for explosives. Compressed gas is discharged beneath a sand layer, simulating the dynamic flow generated by a buried explosive. High speed photography reveals that the stages of soil motion observed during a landmine blast are replicated. The effect of soil saturation and the depth of the sand layer on sand motion are evaluated. Two series of experiments have been performed with the buried charge simulator to characterise subsequent impact of the sand. First, the time variation in pressure and impulse during sand impact on a stationary target is evaluated using a Kolsky bar apparatus. It is found that the pressure pulse imparted to the Kolsky bar consists of two phases: an initial transient phase of high pressure (attributed to wave propagation effects in the impacting sand), followed by a lower pressure phase of longer duration (due to lateral flow of the sand against the Kolsky bar). Both phases make a significant contribution to the total imparted impulse. It is found that wet sand exerts higher peak pressures and imparts a larger total impulse than dry sand. The level of imparted impulse is determined as a function of sand depth, and of stand-off distance between the sand and the impacted end of the Kolsky bar. The second study uses a vertical impulse pendulum to measure the momentum imparted by sand impact to a target which is free to move vertically. The effect of target mass upon imparted momentum is investigated. It is concluded that the laboratory-scale sand impact apparatus is a flexible tool for investigating the interactions between structures and dynamic sand flows. © 2013 Elsevier Ltd. All rights reserved.

Identification of the steering control behaviour of five test subjects following a randomly curving path in a driving simulator

The paper is concerned with the identification of theoretical preview steering controllers using data obtained from five test subjects in a fixed-base driving simulator. An understanding of human steering control behaviour is relevant to the design of autonomous and semi-autonomous vehicle controls. The driving task involved steering a linear vehicle along a randomly curving path. The theoretical steering controllers identified from the data were based on optimal linear preview control. A direct-identification method was used, and the steering controllers were identified so that the predicted steering angle matched as closely as possible the measured steering angle of the test subjects. It was found that identification of the driver's time delay and noise is necessary to avoid bias in identification of the controller parameters. Most subjects' steering behaviour was predicted well by a theoretical controller based on the lateral/yaw dynamics of the vehicle. There was some evidence that an inexperienced driver's steering action was better represented by a controller based on a simpler model of the vehicle dynamics, perhaps reflecting incomplete learning by the driver. Copyright © 2014 Inderscience Enterprises Ltd.

Directional performance in motion transparency

Motion transparency provides a challenging test case for our understanding of how visual motion, and other attributes, are computed and represented in the brain. However, previous studies of visual transparency have used subjective criteria which do not confirm the existence of independent representations of the superimposed motions. We have developed measures of performance in motion transparency that require observers to extract information about two motions jointly, and therefore test the information that is simultaneously represented for each motion. Observers judged whether two motions were at 90 to one another; the base direction was randomized so that neither motion taken alone was informative. The precision of performance was determined by the standard deviations (S.D.s) of probit functions fitted to the data. Observers also made judgments of orthogonal directions between a single motion stream and a line, for one of two transparent motions against a line and for two spatially segregated motions. The data show that direction judgments with transparency can be made with comparable accuracy to segregated (non-transparent) conditions, supporting the idea that transparency involves the equivalent representation of two global motions in the same region. The precision of this joint direction judgment is, however, 2–3 times poorer than that for a single motion stream. The precision in directional judgment for a single stream is reduced only by a factor of about 1.5 by superimposing a second stream. The major effect in performance, therefore, appears to be associated with the need to compute and compare two global representations of motion, rather than with interference between the dot streams per se. Experiment 2tested the transparency of motions separated by a range of angles from 5 to 180 by requiring subjects to set a line matching the perceived direction of each motion. The S.D.s of these settings demonstrated that directions of transparent motions were represented independently for separations over 20. Increasing dot speeds from 1 to 10 deg/s improved directional performance but had no effect on transparency perception. Transparency was also unaffected by variations of density between 0.1 and 19 dots/deg2

Framework for Establishing Limits of Tabular Aerodynamic Models for Flight Dynamics Analysis

This paper describes the use of the Euler equations for the generation and testing of tabular aerodynamic models for flight dynamics analysis. Maneuvers for the AGARD Standard Dynamics Model sharp leading-edge wind-tunnel geometry are considered as a test case. Wind-tunnel data is first used to validate the prediction of static and dynamic coefficients at both low and high angles, featuring complex vortical flow, with good agreement obtained at low to moderate angles of attack. Then the generation of aerodynamic tables is described based on a data fusion approach. Time-optimal maneuvers are generated based on these tables, including level flight trim, pull-ups at constant and varying incidence, and level and 90 degrees turns. The maneuver definition includes the aircraft states and also the control deflections to achieve the motion. The main point of the paper is then to assess the validity of the aerodynamic tables which were used to define the maneuvers. This is done by replaying them, including the control surface motions, through the time accurate computational fluid dynamics code. The resulting forces and moments are compared with the tabular values to assess the presence of inadequately modeled dynamic or unsteady effects. The agreement between the tables and the replay is demonstrated for slow maneuvers. Increasing rate maneuvers show discrepancies which are ascribed to vortical flow hysteresis at the higher rate motions. The framework is suitable for application to more complex viscous flow models, and is powerful for the assessment of the validity of aerodynamics models of the type currently used for studies of flight dynamics.

A Novel Approach for Axle Detection in Bridge Weigh-In-Motion Systems

Bridge weigh-in-motion (B-WIM), a system that uses strain sensors to calculate the weights of trucks passing on bridges overhead, requires accurate axle location and speed information for effective performance. The success of a B-WIM system is dependent upon the accuracy of the axle detection method. It is widely recognised that any form of axle detector on the road surface is not ideal for B-WIM applications as it can cause disruption to the traffic (Ojio & Yamada 2002; Zhao et al. 2005; Chatterjee et al. 2006). Sensors under the bridge, that is Nothing-on-Road (NOR) B-WIM, can perform axle detection via data acquisition systems which can detect a peak in strain as the axle passes. The method is often successful, although not all bridges are suitable for NOR B-WIM due to limitations of the system. Significant research has been carried out to further develop the method and the NOR algorithms, but beam-and-slab bridges with deep beams still present a challenge. With these bridges, the slabs are used for axle detection, but peaks in the slab strains are sensitive to the transverse position of wheels on the beam. This next generation B-WIM research project extends the current B-WIM algorithm to the problem of axle detection and safety, thus overcoming the existing limitations in current state-of–the-art technology. Finite Element Analysis was used to determine the critical locations for axle detecting sensors and the findings were then tested in the field. In this paper, alternative strategies for axle detection were determined using Finite Element analysis and the findings were then tested in the field. The site selected for testing was in Loughbrickland, Northern Ireland, along the A1 corridor connecting the two cities of Belfast and Dublin. The structure is on a central route through the island of Ireland and has a high traffic volume which made it an optimum location for the study. Another huge benefit of the chosen location was its close proximity to a nearby self-operated weigh station. To determine the accuracy of the proposed B-WIM system and develop a knowledge base of the traffic load on the structure, a pavement WIM system was also installed on the northbound lane on the approach to the structure. The bridge structure selected for this B-WIM research comprised of 27 pre-cast prestressed concrete Y4-beams, and a cast in-situ concrete deck. The structure, a newly constructed integral bridge, spans 19 m and has an angle of skew of 22.7°.