Engineering enzyme specificity using computational design of a defined-sequence library

Engineered biosynthetic pathways have the potential to produce high-value molecules from inexpensive feedstocks, but a key limitation is engineering enzymes with high activity and specificity for new reactions. Here, we developed a method for combining structure-based computational protein design with library-based enzyme screening, in which inter-residue correlations favored by the design are encoded into a defined-sequence library. We validated this approach by engineering a glucose 6-oxidase enzyme for use in a proposed pathway to convert D-glucose into D-glucaric acid. The most active variant, identified after only one round of diversification and screening of only 10,000 wells, is approximately 400-fold more active on glucose than is the wild-type enzyme. We anticipate that this strategy will be broadly applicable to the discovery of new enzymes for engineered biological pathways.     

Aspirin attenuates the anti-inflammatory effects of theophylline via inhibition of cAMP production in mice with non-eosinophilic asthma

Theophylline is commonly used to treat severe asthma and chronic obstructive pulmonary disease (COPD) characterized by non-eosinophilic inflammation. Acetyl salicylic acid (ASA) is one of the most widely used medications worldwide, but up to 20% of patients with asthma experience aggravated respiratory symptoms after taking ASA. Here we evaluated the adverse effect of ASA on the therapeutic effect of theophylline in mice with non-eosinophilic asthma. A non-eosinophilic asthma mouse model was induced by airway sensitization with lipopolysaccharide-containing allergen and then challenged with allergen alone. Therapeutic intervention was performed during allergen challenge. Theophylline inhibited lung inflammation partly induced by T_h1 immune response. ASA attenuated the beneficial effects of theophylline. However, co-administration of the ASA metabolite salicylic acid (SA) showed no attenuating effect on theophylline treatment. The therapeutic effect of theophylline was associated with increase in cAMP levels, which was blocked by co-treatment of theophylline and ASA. ASA co-treatment also attenuated the anti-inflammatory effects of a specific phosphodiesterase 4 inhibitor. These results demonstrate that ASA reverses anti-inflammatory effects of theophylline, and that ASA exerts its adverse effects through the inhibition of cAMP production. Our data suggest that ASA reverses lung inflammation in patients taking theophylline, although clinical evidence will be needed.     

A near-infrared fluorescent sensor for detection of cyanide in aqueous solution and its application for bioimaging

A new NIR fluorescent sensor based on an amine-substituted heptamethine cyanine dye displayed a highly selective fluorescence enhancement with cyanide in aqueous solutions, and was applied for the imaging of anthropogenic and biogenic cyanide.     

Swallowtail band structure of the superfluid Fermi gas in an optical lattice

We investigate the energy band structure of the superfluid flow of ultracold dilute Fermi gases in a one-dimensional optical lattice along the BCS to Bose-Einstein condensate (BEC) crossover within a mean-field approach. In each side of the crossover region, a loop structure (swallowtail) appears in the Bloch energy band of the superfluid above a critical value of the interaction strength. The width of the swallowtail is largest near unitarity. Across the critical value of the interaction strength, the profiles of density and pairing field change more drastically in the BCS side than in the BEC side. It is found that along with the appearance of the swallowtail, there exists a narrow band in the quasiparticle energy spectrum close to the chemical potential, and the incompressibility of the Fermi gas consequently experiences a profound dip in the BCS side, unlike in the BEC side.     

Surface redox induced bipolar switching of transition metal oxide films examined by scanning probe microscopy

The bipolar resistive switching mechanisms of a p-type NiO film and n-type TiO₂ film were examined using local probe-based measurements. Scanning probe-based current–voltage (I–V) sweeps and surface potential/current maps obtained after the application of dc bias suggested that resistive switching is caused mainly by the surface redox reactions involving oxygen ions at the tip/oxide interface. This explanation can be applied generally to both p-type and n-type conducting resistive switching films. The contribution of oxygen migration to resistive switching was also observed indirectly, but only in the cases where the tip was in (quasi-) Ohmic contact with the oxide.     

Long distance laser ultrasonic propagation imaging system for damage visualization

Wind turbine blade failure is the most prominent and common type of damage occurring in operating wind turbine systems. Conventional nondestructive testing systems are not available for in situ wind turbine blades. We propose a portable long distance ultrasonic propagation imaging (LUPI) system that uses a laser beam targeting and scanning system to excite, from a long distance, acoustic emission sensors installed in the blade. An examination of the beam collimation effect using geometric parameters of a commercial 2 MW wind turbine provided Lamb wave amplitude increases of 41.5 and 23.1 dB at a distance of 40 m for symmetrical and asymmetrical modes, respectively, in a 2 mm-thick stainless steel plate. With this improvement in signal-to-noise ratio, a feasibility study of damage detection was conducted with a 5 mm-thick composite leading edge specimen. To develop a reliable damage evaluation system, the excitation/sensing technology and the associated damage visualization algorithm are equally important. Hence, our results provide a new platform based on anomalous wave propagation imaging (AWPI) methods with adjacent wave subtraction, reference wave subtraction, reference image subtraction, and the variable time window amplitude mapping method. The advantages and disadvantages of AWPI algorithms are reported in terms of reference data requirements, signal-to-noise ratios, and damage evaluation accuracy. The compactness and portability of the proposed UPI system are also important for in-field applications at wind farms.     

Dual-mode operation of a twisted nematic liquid crystal cell by switching between dynamic and memory modes

We propose a twisted nematic liquid crystal device that can be operated in dynamic or memory mode, based on the information content to be displayed at that time. +90°-twisted and -90°-twisted states are used as two stable states for operation in the memory mode. A vertical electric field is applied to realize gray levels for operation in the dynamic mode. The proposed device has a memory retention time of over a month for the memory mode and a response time of 12 ms for the dynamic mode. Contrast ratios of over 500∶1 can be obtained in both the dynamic and memory modes.     

Ultrawidely tunable single-mode fiber acousto-optic filter

A single-mode fiber (SMF) acousto-optic tunable filter (AOTF) with a tuning range of more than 300 nm is demonstrated. The SMF used in the experiment has a ring of symmetric holes within the cladding, which causes a larger mode-index difference between the first and the second higher-order antisymmetric modes than those of a conventional SMF. As a result, the difference in beatlengths between the core mode and the higher-order modes is highly increased, which makes it possible for the SMF AOTF to exhibit a single resonance peak in the transmission spectrum over the wavelength range of 1.3-1.6 μm for given acoustic frequencies of 3.1-3.8 MHz.