Selection strategy to generate aptamer pairs that bind to distinct sites on protein targets

Many analytical techniques benefit greatly from the use of affinity reagent pairs, wherein each reagent recognizes a discrete binding site on a target. For example, antibody pairs have been widely used to dramatically increase the specificity of enzyme linked immunosorbent assays (ELISA). Nucleic acid-based aptamers offer many advantageous features relative to protein-based affinity reagents, including well-established chemical synthesis, thermostability, and low production cost. However, the generation of suitable aptamer pairs has posed a significant challenge, and few such pairs have been reported to date. To address this important challenge, we present multivalent aptamer isolation systematic evolution of ligands by exponential enrichment (MAI-SELEX), a technique designed for the efficient selection of aptamer pairs. In contrast to conventional selection methods, our method utilizes two selection modules to generate separate aptamer pools that recognize distinct binding sites on a single target. Using MAI-SELEX, we have isolated two groups of 2'-fluoro-modified RNA aptamers that specifically recognize the αV or β3 subunits of integrin αVβ3. These aptamers exhibit low nanomolar affinities for their targets, with minimal cross-reactivity to other closely related integrin homologues. Moreover, we show that these aptamer pairs do not interfere with each other's binding and effectively detect the target even in complex mixtures such as undiluted serum.     

Improved vapor selectivity and stability of localized surface plasmon resonance with a surfactant-coated Au nanoparticles film

Here, we report the use of tetraoctylammonium bromide (TOABr)-coated Au nanoparticles (NPs) for the optical sensing of volatile organic compounds (VOCs). We find that the film responded selectively to the presence of polar and nonpolar vapors by changes in the maximum wavelength (λ(max)) toward higher and lower wavelengths, respectively, as determined by UV-visible spectroscopy. We also observed that the organic coating reorganizes when vapors partition into the film indicated by FT-IR and the film contracts in the presence of water indicated by scanning electron microscopy (SEM). In the present sensor, the metallic Au core serves as the plasmonic signal while the organic coating acts as the receptor material providing vapor selectivity and sensor stability. Correlating changes in (λ(max)) with changes in the refractive index (RI) and nanoparticle-to-nanoparticle separation in the film is important both fundamentally and for improving selectivity in localized surface plasmon resonance (LSPR) sensors.     

Detailed kinetic modeling of the combustion of the four butanol isomers in premixed low-pressure flames

With regard to the importance of butanol as a potential replacement or additive to fossil transportation fuels, a detailed understanding of butanol combustion chemistry is desirable. Routes to different isomers of butanol from biomass are becoming available, and it is known that the structure of fuel molecules can be of crucial importance with respect to the intermediate species pool and the nature and amount of potential pollutants, including regulated air toxics. Quantitative major and intermediate species profiles for the combustion of the four butanol isomers under low-pressure premixed flame conditions, measured with two different in situ mass spectrometric instruments have recently been reported (P. Oßwald et al., Combust. Flame 158 (2011) 2–15), and this large consistent dataset has motivated us to further develop a detailed and comprehensive chemical kinetic model which was used here to complement the experimental results with numerical simulations, including reaction flow analyses. The major differences in the overall chemical pathways in these flames will be discussed, especially with respect to the formation of undesired emissions, and from agreement and differences between experiment and model, suggestions for further work will be given.     

Combination of electrophoretic deposition and microwave-ignited combustion synthesis for the preparation of ceramic coated intermetallic-based materials

Electrophoretic deposition (EPD) was used to deposit sub-micrometric ZrO₂ particles on metallic powder compacts belonging to the systems Ni + Al and Ti + Al, which were used as deposition electrodes in the EPD cell. After EPD, combustion synthesis (CS) of such reactive electrodes was ignited in a microwave single-mode applicator, operating at a frequency of 2.45 GHz, in order to obtain in a single step the synthesis of the desired intermetallic phase (substrate) and the sintering of the previously deposited ceramic particles (or coating). Experimental results demonstrate that the excess heat released during the formation of nickel and titanium aluminides by CS can be exploited not only to self-sustain and self-propagate the reaction front along the substrate, but also to rapidly sinter the coating obtained by EPD. The innovative procedure here proposed is a promising strategy in order to obtain, in a single step, high temperature intermetallic-based materials, protected by well adhered ceramic coatings.     

Effect of aging on microstructural development in an Al–Mg–Si alloy processed by high-pressure torsion

Experiments were conducted to evaluate the microstructural evolution in a commercial Al-0.6 % Mg-0.4 % Si alloy processed using high-pressure torsion for up to 20 turns. Disks of the alloy were tested in two different conditions: in a solution-treated condition and after a short aging treatment at 523 K. The results show that HPT processing introduces significant grain refinement through HPT processing including a reduction in grain size from ~150 μm to ~720 nm in 1 turn of HPT. The final grain size in this alloy was ~250 nm after 20 turns. Some tensile tests were conducted to evaluate the mechanical properties of the alloy at the solution treatment temperature. The results from these tests show that aging at 523 K leads to a small increase in ductility for all tensile samples with a maximum recorded elongation of ~230 %.     

Spin doping of individual molecules by using single-atom manipulation

Being able to control the spin of magnetic molecules at the single-molecule level will make it possible to develop new spin-based nanotechnologies. Gate-field effects and electron and photon excitations have been used to achieve spin switching in molecules. Here, we show that atomic doping of molecules can be used to change the molecular spin. Furthermore, a scanning tunneling microscope was used to place or remove the atomic dopant on the molecule, allowing us to change the molecular spin in a controlled way. Bis(phthalocyaninato)yttrium (YPc(2)) molecules deposited on an Au (111) surface keep their spin-1/2 magnetic moment due to the small molecule-substrate interaction. However, when Cs atoms were carefully placed onto YPc(2) molecules, the spin of the molecule vanished as shown by our conductance measurements and corroborated by the results of density functional theory calculations.     

Plasmonic nanosensors with inverse sensitivity by means of enzyme-guided crystal growth

Lowering the limit of detection is key to the design of sensors needed for food safety regulations, environmental policies and the diagnosis of severe diseases. However, because conventional transducers generate a signal that is directly proportional to the concentration of the target molecule, ultralow concentrations of the molecule result in variations in the physical properties of the sensor that are tiny, and therefore difficult to detect with confidence. Here we present a signal-generation mechanism that redefines the limit of detection of nanoparticle sensors by inducing a signal that is larger when the target molecule is less concentrated. The key step to achieve this inverse sensitivity is to use an enzyme that controls the rate of nucleation of silver nanocrystals on plasmonic transducers. We demonstrate the outstanding sensitivity and robustness of this approach by detecting the cancer biomarker prostate-specific antigen down to 10(-18)?g?ml(-1) (4 × 10(-20)?M) in whole serum.     

Influence of Pressing Temperature on Microstructure Evolution and Mechanical Behavior of Ultrafine‐Grained Cu Processed by Equal‐Channel Angular Pressing

Pure Cu was processed by ECAP at five different temperatures from room temperature (RT) to 523 K. The influence of pressing temperature on microstructure evolution and tensile behavior was investigated in detail. The results show that as the ECAP temperature is increased the grain size and ductility both increase whereas the dislocation density and yield strength decrease. In the case of ECAP processing in the range of RT to 473 K the mechanism governing microstructural refinement is continuous dynamic recrystallization (CDRX), whereas at 523 K the mechanism changes to discontinuous dynamic recrystallization (DDRX). At higher ECAP temperatures, the kinetics of CDRX are retarded leading to a lower fraction of equiaxed grains/high‐angle grain boundaries and a higher fraction of dislocation cell structures. At 523 K, DDRX induces a high fraction of equiaxed grains with a very low dislocation density which appears responsible for the observed high tensile ductility. The sample processed at 523 K possessed a good combination of strength and ductility, suggesting that processing by ECAP at elevated temperatures may be a suitable alternative to RT ECAP processing followed by subsequent annealing.     

A novel bioactive vitroceramic presents similar biological responses as autogenous bone grafts

Bioactive glasses represent an interesting class of bone substitute's biomaterials. The present study investigated the repair of bone defects filled with a novel bioactive vitroceramic (Biosilicate(?)), alone or in association with particulate autogenous bone grafts in calvaria defects of rabbits. After 7, 14, and 30 days the specimens were retrieved for histological, histomorphometric and immunohistochemistry analysis. Satisfactory bone formation was observed in all groups, and direct bone-biomaterial surface was noted. Histomorphometric assessment did not show statistically significant differences in bone formation among the groups and periods, except for BG group at day 14. Immunoexpression of Runx-2 was similar among the groups containing the graft and the biomaterial, being more intense than in control group. Similar result was observed for VEGF expression, especially in the last experimental period. These results revealed that Biosilicate(?) presented a favorable behavior, comparable to autogenous bone graft.