In Vivo Delivery and Activation of Masked Fluorogenic Hydrolase Substrates by Endogenous Hydrolases in C. elegans

Protein expression and localization are often studied in vivo by tagging molecules with green fluorescent protein (GFP), yet subtle changes in protein levels are not easily detected. To develop a sensitive in vivo method to amplify fluorescence signals and allow cell‐specific quantification of protein abundance changes, we sought to apply an enzyme‐activated cellular fluorescence system in vivo by delivering ester‐masked fluorophores to Caenorhabditis elegans neurons expressing porcine liver esterase (PLE). To aid uptake into sensory neuron membranes, we synthesized two novel fluorogenic hydrolase substrates with long hydrocarbon tails. Recombinant PLE activated these fluorophores in vitro. In vivo activation occurred in sensory neurons, along with potent activation in intestinal lysosomes quantifiable by imaging and microplate and partially attributable to gut esterase 1 (GES‐1) activity. These data demonstrate the promise of biorthogonal hydrolases and their fluorogenic substrates as in vivo neuronal imaging tools and for characterizing endogenous C. elegans hydrolase substrate specificities.     

Impact of Relative Humidity on HVAC Filters Loaded with Hygroscopic and Non-Hygroscopic Particles

The key characteristics of an air filter—flow resistance and filtration efficiency—are strongly affected by captured particles. The impact of exposing loaded heating, ventilating, and air conditioning air filters to a relative humidity (RH) other than that experienced during loading is investigated to develop an understanding of the role of RH throughout filter operation. Flat sheets of commercial filter media were loaded with hygroscopic, non-hygroscopic, or a mixture of particles in a laboratory apparatus. When filters loaded with hygroscopic particles in dry air were exposed to an elevated RH of 40%, the flow resistance reduced by up to 47%, depending on the filter being tested. Investigation of filter efficiency before and after changes in RH in the same samples shows reductions of up to 11 percentage points in the 130-nm size range. Further increasing RH causes additional drops in flow resistance and efficiency whereas reverting back to a lower humidity does not change the filter characteristics. The irreversibility of the particle-loaded filter characteristics implies that the RH increases are associated with an irreversible change in the particle structure. The response to humidity was reduced if an aerosol mixture of hygroscopic and non-hygroscopic particles is used. Exposure of filters loaded with only non-hygroscopic particles does not show the same dependence on RH. Small increases in growth factor for RH changes below deliquescence, causing morphological changes in captured particle aggregates, is a potential explanation for the changes observed.

Copyright 2015 American Association for Aerosol Research     

Sintering mechanisms of Yttria with different additives

The sintering behaviour of conventional yttria powder was investigated, with emphasis on the effect of sintering additives such as B₂O₃, YF₃, Al₂O₃, ZrO₂, and TiO₂, etc. at sintering temperatures from 1000 °C to 1600 °C. Powder shrinkage behaviour was analysed using a dilatometer. The powder sintering mechanisms were identified at different temperatures using powder isothermal shrinkage curves. This analysis showed that the sintering additives B₂O₃ and YF₃ could improve yttria sintering by changing the diffusion/sintering mechanisms at certain temperatures, while sintering additives TiO₂, Al₂O₃ and ZrO₂ appeared to retard the powder densification at temperatures around 1000 °C and are more suitable when used at temperatures in excess of 1300 °C. The powder with La₂O₃ added had the slowest densification rate throughout the test temperatures in this experiment and was also found to be more suitable when used at temperatures higher than 1550 °C.     

Native state kinetic stabilization as a strategy to ameliorate protein misfolding diseases: a focus on the transthyretin amyloidoses

Small molecule-mediated protein stabilization inside or outside of the cell is a promising strategy to treat protein misfolding/misassembly diseases. Herein we focus on the transthyretin (TTR) amyloidoses and demonstrate that preferential ligand binding to and stabilization of the native state over the dissociative transition state raises the kinetic barrier of dissociation (rate-limiting for amyloidogenesis), slowing and in many cases preventing TTR amyloid fibril formation. Since T119M-TTR subunit incorporation into tetramers otherwise composed of disease-associated subunits also imparts kinetic stability on the tetramer and ameliorates amyloidosis in humans, it is likely that small molecule-mediated native state kinetic stabilization will also alleviate TTR amyloidoses.     

Methane partial oxidation to synthesis gas over bimetallic cobalt/tungsten carbide catalysts and integration with a Mn substituted hexaaluminate combustion catalyst

Co_0.2W_0.8C_x and supported Co_0.2W_0.8C_x catalysts are shown to be active for the partial oxidation of methane to synthesis gas. The catalyst stability is improved by operating at elevated pressure, or in the presence of excess methane. At atmospheric pressure the Co_0.2W_0.8C_x catalysts deactivate by oxidation, as seen by X-ray diffraction. Manganese substituted hexaaluminate catalysts with different Mn contents have been tested as catalysts for the total combustion of methane. In particular BaMn₂Al₁₀O₁₉ is active and stable for the combustion reaction. The temperature rise observed in the reactor was up to 300 K, depending on the reaction conditions, and complete conversion of oxygen in the feed was achieved. A process for stabilising the carbide catalysts is demonstrated, combining the manganese substituted hexaaluminate total oxidation catalyst, in series before the carbide reforming catalyst: this process leads to stable operation, with no carbon formation in the reactor and no carbide catalyst oxidation observed.     

Elastic Indentation Response of Float Glass Surfaces

Instrumented Hertzian indentation was used to evaluate the reduced elastic modulus and cone-crack initiation forces for the as-received surfaces of commercial float glasses. Custom-built indentation equipment with the capability of acoustic emission detection was used to monitor continuously the load and depth of penetration at the microscopic scale for forces up to 1 kN. Equipment verification was performed using a reference material, GE 124. The air and tin surfaces of commercial soda–lime–silica and borosilicate float glasses were tested to determine any difference in indentation response for the elastic and fracture behavior of as-received surfaces. Information obtained from the analysis of the load–displacement curves and from the visual inspection of the indentation sites was used to determine the elastic modulus, and the conditions for the onset of cone cracking as a function of surface roughness. The reduced modulus results were verified using additional equipment that allowed the in situ observation of the contact area during loading and unloading. The results showed that there was no difference in the reduced modulus data for the air and tin surfaces for the range of surface displacements studied. The same conclusions were found for cone-cracking loads on as-received surfaces but tests on abraded surfaces showed that the tin surfaces had slightly more resistance to cone cracking than the air surfaces.     

Thermal Shock Resistance of Ceramic Foams

Thermal shock behavior of a variety of open-cell ceramic foams was evaluated using infrared heating and forced air cooling. The extent of damage after thermal shock was determined by a nondestructive, dynamic resonance technique. The damage in foams was found to be strongly dependent on cell size and weakly dependent on density. In zirconia-based foams, damage was found to increase with an increase in zirconia content. A thermal stress resistance parameter R '_f was derived to predict the effect of cell size and density on the damage incurred in foams. The experimental results were found to corroborate the predictions fairly well but a better approach was to compare the maximum applied thermal strains with the degree of damage.     

Indentation Determination of Fatigue Limits in Silicate Glasses

An experimental approach has been used to measure the threshold stress intensity factor ( K _th) for subcritical crack growth in brittle materials using indentation cracks. The data show that K _th values existed in soda-lime-silica and soda-alumina-silica glasses that were tested in a neutral aqueous environment. For the former glass, tests also were performed in acidic and basic solutions. A threshold for subcritical crack growth was observed for acidic conditions but not for alkaline conditions.     

Using a conductive sphere as a probe to characterize the sensitivity of soft piezoresistive films

Flexible piezoresistive films, such as, carbon black/polydimethylsiloxane (C-PDMS) composites, are often used as skin analogs and integrated into complex array sensors for tactile sensing. The uniformity of the sensor characteristics heavily depends on the homogeneity of the composite. Therefore, the ability to locally characterize a film that will be integrated into a complex force sensor could be critical. Here, a method to characterize the local sensitivity of flexible piezoresistive films is presented. Using a conductive sphere, which was chosen over a flat probe to eliminate misalignment issues, the surface of a thin film composite is indented to characterize the change in resistivity in terms of average strain. Experiments were performed with 15 and 18 wt% carbon black C-PDMS films of varying thickness. The contact radius of the probe with the piezoresistive film was estimated using the Johnson-Roberts-Kendall contact theory. Theoretical contact area estimates were found to agree with contact radius measurements carried out using optically transparent PDMS films observed through an optical microscope. Results show that C-PDMS with 15 wt% carbon black exhibit a higher rate if change of resistivity and gauge factor than films of same thickness with 18 wt% carbon black. On the other hand, thicker films exhibit higher gauge factors for the two tested carbon black contents. Tests carried out at multiple locations yielded consistent sensitivity values, making these types of composites suitable for array type force sensors.