Site-Specific Isotopic Labeling (SSIL): Access to High-Resolution Structural and Dynamic Information in Low-Complexity Proteins

Remarkable technical progress in the area of structural biology has paved the way to study previously inaccessible targets. For example, large protein complexes can now be easily investigated by cryo-electron microscopy, and modern high-field NMR magnets have challenged the limits of high-resolution characterization of proteins in solution. However, the structural and dynamic characteristics of certain proteins with important functions still cannot be probed by conventional methods. These proteins in question contain low-complexity regions (LCRs), compositionally biased sequences where only a limited number of amino acids is repeated multiple times, which hamper their characterization. This Concept article describes a site-specific isotopic labeling (SSIL) strategy, which combines nonsense suppression and cell-free protein synthesis to overcome these limitations. An overview on how poly-glutamine tracts were made amenable to high-resolution structural studies is used to illustrate the usefulness of SSIL. Furthermore, we discuss the potential of this methodology to give further insights into the roles of LCRs in human pathologies and liquid–liquid phase separation, as well as the challenges that must be addressed in the future for the popularization of SSIL.     

Macrocell corrosion of steel in concrete: Characterization of anodic behavior in relation to the chloride content

This study presents the determination of electrochemical properties of active steel in mortar, based on inverse numerical modeling that focuses on their dependency on chloride content. An experimental campaign, consisting of galvanic coupling tests between anode samples contaminated with different chloride concentrations and cathode samples without chlorides, was carried out. Cathode polarization tests allowed for directly determining passive steel electrochemical parameters. Anode polarization tests coupled with a numerical optimization were then performed for quantifying active steel parameters and focusing on chloride's effect on the iron anodic Tafel coefficient. Furthermore, the steel electrochemical properties were successfully used as input parameters to model the galvanic experiments.     

Collision Avoidance Using a Cerebellar Model Arithmetic Computer Neural Network

Avoiding collisions with obstacles in a clustered environment is a difficult task for autonomous vehicles. Deterministic algorithms cannot address all scenarios encountered and may fail to perform in dynamically changing environments. Neural networks, owing to their ability to map complex relationships between multiple input-output patterns, can learn the task of maneuvering around and in-between obstacles to reach a goal state. The Cerebellar Model Arithmetic Computer (CMAC) neural network in particular is based on a model of human sensory motor responses and can efficiently model responsive control actions. A CMAC neural network controller was developed and examined, in simulation, for its suitability to capture a driver's function of steering and braking. The performance of the controller was tested in a simulation of a moving platform (vehicle) encountering obstacles of various shapes, whereas the CMAC was trained only with limited shapes and scenarios. Preliminary simulation results have shown the CMAC's ability to successfully generalize its learned patterns to avoid obstacles after only a few training sessions. The CMAC output is generated in a computationally efficient manner with physically and economically realizable memory sizes. Therefore, real-time hardware implementation of the controller is feasible. This research demonstrates that the method has the ability to accommodate more complex scenarios.     

Theory of the universal and nonuniversal quantities of fluids at the critical point

We show that the hierarchical reference theory is an accurate global theory of fluids at least above the critical temperatureT _c. The hierarchy is truncated at the first equation, the one connecting the free energy to the pair correlation function, with an Ornstein-Zernike ansatz. In this approximation the theory can be considered as a sophisticated generalization of the optimized random phase approximation which has genuine nonclassical critical exponents and for which scaling is satisfied. We study the system of hard spheres plus the Lennard-Jones attractive well and find a good agreement with measuredPVT, specific heat, correlation length, and structure factor in rare gases. The accuracy of the theory remains very good up to freezing density.Paper presented at the Tenth Symposium on Thermophysical Properties, June 20–23, 1988, Gaithersburg, Maryland, U.S.A.     

Evidence for the covalent binding of hydroxyethyl radicals to rat liver microsomal proteins

It is well known that acetaldehyde is capable of covalent binding to liver proteins. However, in experiments using liver microsomes prepared from chronically ethanol-fed rats we have observed that the addition of EDTA-iron complex to the microsomes increases by about 4-5 fold both the spin trapping of hydroxyethyl radicals and the covalent binding of 14C-ethanol to proteins, while it only doubles acetaldehyde formation. Conversely, the presence of GSH strongly decreases the trapping of hydroxyethyl radicals and completely inhibits the covalent binding, without affecting acetaldehyde production. Furthermore, the spin trapping agent 4-pyridyl-N-oxide-t-butyl nitrone (4-POBN), previously employed for the detection of hydroxyethyl radicals, decreases by about 70% the covalent binding of 14C-ethanol to microsomal proteins. 4-POBN does not affect acetaldehyde production by liver microsomes, nor does it interfere with the covalent binding of acetaldehyde produced by ADH-mediated oxidation of ethanol. The results obtained indicate that hydroxyethyl radicals generated during ethanol oxidation by cytochrome P-450 play an important role in the alkylation of microsomal proteins consequent to ethanol metabolism.     

Wireless Dual Stimuli Actuation of Dye Sensitized Conducting Polymer Hybrids

Actuators controlled by external stimuli have received a lot of attention in recent years. Herein a polymer based dual stimuli actuator is reported, triggered by light and an electric field. This allows better control of actuation, enlarging the field of potential applications, like, for example, in the frame of soft robotics. The actuator is composed of polypyrrole and TiO₂ modified with methylene blue. In an aqueous solution, the resulting freestanding hybrid film shows reversible actuation due to the synergy of light and an applied electric field. Illumination with light produces electron-hole pairs in the TiO₂ layer, which are shuttled to the opposite ends of the actuator by the potential gradient present in the solution. This results in electrochemical oxidation and reduction reactions at the two extremities and consequently in site selective swelling of the polymer, which finally leads to a controlled motion of the actuator, following the principles of logic gate operations. Such synergistically induced switching allows developing original actuation schemes for performing complex mechanical tasks triggered by more than one stimulus.     

CoMo catalysts supported on aluminosilicates: synergy between support and sodium effects

The structural properties and the hydrodesulfurization (HDS) activity of sodium doped and sodium free CoMo catalysts supported on amorphous aluminosilicates (ASA) were investigated as a function of different SiO₂:Al₂O₃ ratios. The support yielding the most active catalyst, (66% alumina) doped with different amounts of sodium, was used for a series of similar catalysts in order to study the effect of the alkali ion on the catalytic performance. The supports were prepared by sol–gel method and the catalysts were prepared by incipient wet impregnation method. The structure and the surface of the various samples were investigated by X-ray diffraction (XRD), temperature-programmed reduction (TPR) and X-ray photoelectron spectroscopy (XPS). The catalytic behaviour was tested in the hydrodesulfurization of thiophene carried out in a continuous flow system at atmospheric pressure, in a range of temperature between 603 and 633 K. Changes of activity with the support composition were observed. The presence of sodium, modifying the Brønsted acidity of the supports, enhances such effect. Moreover, the increase of the activity with increasing amount of sodium was a clear indication of the promoting effect of the alkali ion.