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.

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.

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.

Initial investigation into the use of GGBS-MgO in soil stabilisation

Portland cement (PC) is the most widely used binder for ground improvement. However, there are significant environmental impacts associated with its production in terms of high energy consumption and CO2 emissions. Hence, the use of industrial by-products materials or new low-carbon footprint alternative cements has been encouraged. Ground granulated blastfurnace slag (GGBS), a by-product of the steel industry, has been successfully used for such an application, usually activated with an alkali such as lime or PC. In this study the use of MgO as a novel activator for GGBS in ground improvement of soft soils is addressed and its performance was compared to the above two conventional activators as well as PC alone. The GGBS:activator ratio used in this study was 9:1. A range of tests was performed at three curing periods (7, 28 and 90 days), including unconfined compressive strength (UCS), permeability and microstructure analysis. The results show that the MgO performed as the most efficient activator yielding the highest strength and the lowest permeability indicating a very high stabilisation efficiency of the system. © 2012 American Society of Civil Engineers.

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.

Using drift nets to capture early life stages and monitor spawning of the Yangtze River Chinese sturgeon (Acipenser sinensis)

P>A sampling system for capturing sturgeon eggs using a D-shaped bottom anchored drift net was used to capture early life stages (ELS) of Chinese sturgeon, Acipenser sinensis, and monitor annual spawning success at Yichang on the Yangtze River, 1996-2004, before and just after the Three Gorges Dam began operation. Captured were 96 875 ELS (early life stages: eggs, yolk-sac larvae = eleuthero embryos, and larvae); most were eggs and only 2477 were yolk-sac larvae. Most ELS were captured in the main river channel and inside the bend at the Yichang spawning reach. Yolk-sac larvae were captured for a maximum of 3 days after hatching began, indicating quick dispersal downstream. The back-calculated day of egg fertilization over the eight years indicated a maximum spawning window of 23 days (20 October-10 November). Spawning in all years was restricted temporally, occurred mostly at night and during one or two spawning periods, each lasting several days. The brief temporal spawning window may reduce egg predation by opportunistic predators by flooding the river bottom with millions of eggs. During 1996-2002, the percentage of fertilized eggs in an annual 20-egg sample was between 63.5 to 94.1%; however, in 2003 the percentage fertilized was only 23.8%. This sudden decline may be related to the altered environmental conditions at Yichang caused by operation of the Three Gorges Dam. Further studies are needed to monitor spawning and changes in egg fertilization in this threatened population.

Comparative responses in rare minnow exposed to 17 beta-estradiol during different life stages

Present in the excrement of humans and animals, 17 beta-estradiol (E-2) has been detected in the aquatic environment in a range from several nanograms to several hundred nanograms per liter. In this study, the sensitivities of rare minnows during different life stages to E-2 at environmentally relevant (5, 25, and 100 ng l(-1)) and high (1000 ng l(-1)) concentrations were compared using vitellogenin (VTG) and gonad development as biomarkers under semistatic conditions. After 21 days of exposure, VTG concentrations in whole-body homogenates were analyzed; the results indicated that the lowest observed effective concentration for VTG induction was 25 ng l(-1) E-2 in the adult stage, but 100 ng l(-1) E-2 in the larval and juvenile stages. After exposure in the early life stage, the larval and juvenile fish were transferred to clean water until gonad maturation. No significant difference in VTG induction was found between the exposure and control groups in the adults. However, a markedly increased proportion of females and appearance of hermaphrodism were observed in the juvenile-stage group exposed to 25 ng l(-1) E-2. These results showed that VTG induction in the adult stage is more sensitive than in larval and juvenile stages following exposure to E-2. The juvenile stage may be the critical period of gonad development. Sex ratio could be a sensitive biomarker indicating exposure to xenoestrogens in early-life-stage subchronic exposure tests. The results of this study provide useful information for selecting sensitive biomarkers properly in aquatic toxicology testing.

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.

Zebraflsh z-otu, a novel Otu and Tudor domain-containing gene, is expressed in early stages of oogenesis and embryogenesis

Several studies have suggested that Otu domain had de-ubiquitinating activity and Tudor domain was important for the formation of germ cells. Here, we reported a novel zebrafish ovary-specific gene containing Otu and Tudor domain, z-otu, which was expressed at stages I-III oocytes and embryonic stages from zygotes to early blastula during embryonic cells maintained their totipotency. Therefore, z-otu might link the ubiquitin signaling pathway to early oogenesis and maintaining the totipotency of embryonic cell. (c) 2005 Elsevier B.V. All rights reserved.

Long-term toxic impact of 2,3,7,8-tetrachlorodibenzo-p-dioxin on the reproduction, sexual differentiation, and development of different life stages of Gobiocypris rarus and Daphnia magna

The sexual ratio of Gobiocypris rarus exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and 17 beta -estradiol from embryo to sexually mature revealed feminization and overdevelopment of connective tissue in male fish gonad in 2-30 pg/L TCDD concentration range. Daphnia magna was not sensitive to the high dose of TCDD (0.1-1000 ng/ml), but the reproduction of D. magna treated with TCDD decreased after the 8th day. 7-Ethoxyresorufin-O-deethylase (EROD) activities in newly fertilized eggs of G. rarus exposed to TCDD dosage groups (1000-100,000 pg/L) were significantly induced and increased with TCDD concentrations at the early life stage, while no difference was found between low TCDD dosage groups (<100 pg/L), but a good relationship between the EROD activity and the TCDD concentration was observed during a long-term developmental stage. There was a pericardial edema formed in a 2-week yolk-sac at the concentration of 1000 pg/L TCDD. But in the exposure group (2 pg/L TCDD for 120 days), the cell nuclei of hepatocytes was far from the center and packed toward the cell membrane; the cristae of most mitochondria in the cell dropped and collapsed; the rough endoplasmic reticulum broke into fragments; and numerous lipid droplets formed in the cell. (C) 2001 Academic Press.

The influence of growth temperature and input V/III ratio on the initial nucleation and material properties of InN on GaN by MOCVD

The effects of growth temperature and V/III ratio on the InN initial nucleation of islands on the GaN (0 0 0 1) surface were investigated. It is found that InN nuclei density increases with decreasing growth temperature between 375 and 525 degrees C. At lower growth temperatures, InN thin films take the form of small and closely packed islands with diameters of less than 100 nm, whereas at elevated temperatures the InN islands can grow larger and well separated, approaching an equilibrium hexagonal shape due to enhanced surface diffusion of adatoms. At a given growth temperature of 500 degrees C, a controllable density and size of separated InN islands can be achieved by adjusting the V/III ratio. The larger islands lead to fewer defects when they are coalesced. Comparatively, the electrical properties of the films grown under higher V/III ratio are improved.

Study on growth mechanism of low-temperature prepared microcrystalline Si thin films

Microcrystalline silicon thin films at different growth stages were prepared by hot wire chemical vapor deposition. Atomic force microscopy has been applied to investigate the evolution of surface topography of these films. According to the fractal analysis I it was found that, the growth of Si film deposited on glass substrate is the zero-diffused stochastic deposition; while for the film on Si substrate, it is the finite diffused deposition on the initial growth stage, and transforms to the zero-diffused stochastic deposition when the film thickness reaches a certain value. The film thickness dependence of island density shows that a maximum of island density appears at the critical film thickness for both substrates. The data of Raman spectra approve that, on the glass substrate, the a-Si: H/mu c-Si:H transition is related to the critical film thickness. Different substrate materials directly affect the surface diffusion ability of radicals, resulting in the difference of growth modes on the earlier growth stage.

The influence of V/III ratio in the initial growth stage on the properties of GaN epilayer deposited on low temperature AlN buffer layer

The V/III ratio in the initial growth stage of metalorganic chemical vapor deposition has an important influence on the quality of a GaN epilayer grown on a low-temperature AIN buffer layer and c-plane sapphire substrate. A weaker yellow luminescence, a narrower half-width of the X-ray diffraction peak, and a higher electron mobility result when a lower V/III ratio is taken. The intensity of in situ optical reflectivity measurements indicates that the film surface is rougher at the beginning of GaN growth, and a longer time is needed for the islands to coalesce and for a quasi-two dimensional mode growth to start. A comparison of front- and back-illuminated photoluminescence spectra confirms that many threading dislocations are bent during the initial stage, leading to a better structural quality of the GaN layer. (C) 2007 Elsevier B.V. All rights reserved.