Nanobodies raised against monomeric α-synuclein distinguish between fibrils at different maturation stages

Nanobodies are single-domain fragments of camelid antibodies that are emerging as versatile tools in biotechnology. We describe here the interactions of a specific nanobody, NbSyn87, with the monomeric and fibrillar forms of α-synuclein (αSyn), a 140-residue protein whose aggregation is associated with Parkinson's disease. We have characterized these interactions using a range of biophysical techniques, including nuclear magnetic resonance and circular dichroism spectroscopy, isothermal titration calorimetry and quartz crystal microbalance measurements. In addition, we have compared the results with those that we have reported previously for a different nanobody, NbSyn2, also raised against monomeric αSyn. This comparison indicates that NbSyn87 and NbSyn2 bind with nanomolar affinity to distinctive epitopes within the C-terminal domain of soluble αSyn, comprising approximately amino acids 118-131 and 137-140, respectively. The calorimetric and quartz crystal microbalance data indicate that the epitopes of both nanobodies are still accessible when αSyn converts into its fibrillar structure. The apparent affinities and other thermodynamic parameters defining the binding between the nanobody and the fibrils, however, vary significantly with the length of time that the process of fibril formation has been allowed to progress and with the conditions under which formation occurs, indicating that the environment of the C-terminal domain of αSyn changes as fibril assembly takes place. These results demonstrate that nanobodies are able to target forms of potentially pathogenic aggregates that differ from each other in relatively minor details of their structure, such as those associated with fibril maturation. © 2013 Elsevier Ltd.

Aerodynamic design of high end wall angle turbine stages-part I: Methodology development

A design methodology is presented for turbines in an annulus with high end wall angles. Such stages occur where large radial offsets between the stage inlet and stage outlet are required, for example in the first stage of modern low pressure turbines, and are becoming more prevalent as bypass ratios increase. The turbine vanes operate within s-shaped ducts which result in meridional curvature being of a similar magnitude to the bladeto-blade curvature. Through a systematic series of idealized computational cases, the importance of two aspects of vane design are shown. First, the region of peak end wall meridional curvature is best located within the vane row. Second, the vane should be leant so as to minimize spanwise variations in surface pressure-this condition is termed "ideal lean." This design philosophy is applied to the first stage of a low pressure turbine with high end wall angles. © 2014 by ASME.

Aerodynamic design of high end wall angle turbine stages- part II: Experimental verification

An experimental investigation of a turbine stage featuring very high end wall angles is presented. The initial turbine design did not achieve a satisfactory performance and the difference between the design predictions and the test results was traced to a large separated region on the rear suction-surface. To improve the agreement between computational fluid dynamics (CFD) and experiment, it was found necessary to modify the turbulence modeling employed. The modified CFD code was then used to redesign the vane, and the changes made are described. When tested, the performance of the redesigned vane was found to have much closer agreement with the predictions than the initial vane. Finally, the flowfield and performance of the redesigned stage are compared to a similar turbine, designed to perform the same duty, which lies in an annulus of moderate end wall angles. A reduction in stage efficiency of at least 2.4% was estimated for the very high end wall angle design. © 2014 by ASME.

Influence of film cooling hole angles and geometries on aerodynamic loss and net heat flux reduction

Turbine design engineers have to ensure that film cooling can provide sufficient protection to turbine blades from the hot mainstream gas, while keeping the losses low. Film cooling hole design parameters include inclination angle (a), compound angle (b), hole inlet geometry, and hole exit geometry. The influence of these parameters on aerodynamic loss and net heat flux reduction is investigated, with loss being the primary focus. Low-speed flat plate experiments have been conducted at momentum flux ratios of IR=0.16, 0.64, and 1.44. The film cooling aerodynamic mixing loss, generated by the mixing of mainstream and coolant, can be quantified using a three-dimensional analytical model that has been previously reported by the authors. The model suggests that for the same flow conditions, the aerodynamic mixing loss is the same for holes with different a and b but with the same angle between the mainstream and coolant flow directions (angle k). This relationship is assessed through experiments by testing two sets of cylindrical holes with different a and b: one set with k=35 deg, and another set with k=60 deg. The data confirm the stated relationship between α, β, k and the aerodynamic mixing loss. The results show that the designer should minimize k to obtain the lowest loss, but maximize b to achieve the best heat transfer performance. A suggestion on improving the loss model is also given. Five different hole geometries (α=35.0 deg, β=0 deg) were also tested: cylindrical hole, trenched hole, fan-shaped hole, D-Fan, and SD-Fan. The D-Fan and the SD-Fan have similar hole exits to the fan-shaped hole but their hole inlets are laterally expanded. The external mixing loss and the loss generated inside the hole are compared. It was found that the D-Fan and the SD-Fan have the lowest loss. This is attributed to their laterally expanded hole inlets, which lead to significant reduction in the loss generated inside the holes. As a result, the loss of these geometries is≈50% of the loss of the fan-shaped hole at IR=0.64 and 1.44. © 2013 by ASME.

A novel theoretical approach to passive control of thermo-acoustic oscillations: Application to ducted heat sources

In this paper, we develop a linear technique that predicts how the stability of a thermo-acoustic system changes due to the action of a generic passive feedback device or a generic change in the base state. From this, one can calculate the passive device or base state change that most stabilizes the system. This theoretical framework, based on adjoint equations, is applied to two types of Rijke tube. The first contains an electrically-heated hot wire and the second contains a diffusion flame. Both heat sources are assumed to be compact so that the acoustic and heat release models can be decoupled. We find that the most effective passive control device is an adiabatic mesh placed at the downstream end of the Rijke tube. We also investigate the effects of a second hot wire and a local variation of the cross-sectional area but find that both affect the frequency more than the growth rate. This application of adjoint sensitivity analysis opens up new possibilities for the passive control of thermo-acoustic oscillations. For example, the influence of base state changes can be combined with other constraints, such as that the total heat release rate remains constant, in order to show how an unstable thermo-acoustic system should be changed in order to make it stable. Copyright © 2013 by ASME.

Transient ventilation dynamics induced by heat sources of unequal strength

We examine theoretically the transient displacement flow and density stratification that develops within a ventilated box after two localized floor-level heat sources of unequal strengths are activated. The heat input is represented by two non-interacting turbulent axisymmetric plumes of constant buoyancy fluxes B1 and B2 > B1. The box connects to an unbounded quiescent external environment of uniform density via openings at the top and base. A theoretical model is developed to predict the time evolution of the dimensionless depths λj and mean buoyancies δj of the 'intermediate' (j = 1) and 'top' (j = 2) layers leading to steady state. The flow behaviour is classified in terms of a stratification parameter S, a dimensionless measure of the relative forcing strengths of the two buoyant layers that drive the flow. We find that dδ1/dτ α 1/λ1 and dδ2/dτ α 1/λ2, where τ is a dimensionless time. When S 1, the intermediate layer is shallow (small λ1), whereas the top layer is relatively deep (large λ2) and, in this limit, δ1 and δ2 evolve on two characteristically different time scales. This produces a time lag and gives rise to a 'thermal overshoot', during which δ1 exceeds its steady value and attains a maximum during the transients; a flow feature we refer to, in the context of a ventilated room, as 'localized overheating'. For a given source strength ratio ψ = B1/B2, we show that thermal overshoots are realized for dimensionless opening areas A < Aoh and are strongly dependent on the time history of the flow. We establish the region of {A, ψ} space where rapid development of δ1 results in δ1 > δ2, giving rise to a bulk overturning of the buoyant layers. Finally, some implications of these results, specifically to the ventilation of a room, are discussed. © Cambridge University Press 2013.

On kinematic relaxation and deposition of water droplets in the last stages of low pressure steam turbines

In the first part of the paper steady two-phase flow predictions have been performed for the last stage of a model steam turbine to examine the influence of drag between condensed fog droplets and the continuous vapour phase. In general, droplets due to homogeneous condensation are small and thus kinematic relaxation provides only a minor contribution to the wetness losses. Different droplet size distributions have been investigated to estimate at which size inter-phase friction becomes more important. The second part of the paper deals with the deposition of fog droplets on stator blades. Results from several references are repeated to introduce the two main deposition mechanisms which are inertia and turbulent diffusion. Extensive postprocessing routines have been programmed to calculate droplet deposition due to these effects for a last stage stator blade in three-dimensions. In principle the method to determine droplet deposition by turbulent diffusion equates to that of Yau and Young [1] and the advantages and disadvantages of this relatively simple method are discussed. The investigation includes the influence of different droplet sizes on droplet deposition rates and shows that for small fog droplets turbulent diffusion is the main deposition mechanism. If the droplets size is increased inertial effects become more and more important and for droplets around 1 μm inertial deposition dominates. Assuming realistic droplet sizes the overall deposition equates to about 1% to 3% of the incoming wetness for the investigated guide vane at normal operating conditions. Copyright © 2013 by Solar Turbines Incorporated.