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.

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.