Protease Substrate-Independent Universal Assay for Monitoring Digestion of Native Unmodified Proteins

Proteases are a group of enzymes with a catalytic function to hydrolyze peptide bonds of proteins. Proteases regulate the activity, signaling mechanism, fate, and localization of many proteins, and their dysregulation is associated with various pathological conditions. Proteases have been identified as biomarkers and potential therapeutic targets for multiple diseases, such as acquired immunodeficiency syndrome, cardiovascular diseases, osteoporosis, type 2 diabetes, and cancer, where they are essential to disease progression. Thus, protease inhibitors and inhibitor-like molecules are interesting drug candidates. To study proteases and their substrates and inhibitors, simple, rapid, and sensitive protease activity assays are needed. Existing fluorescence-based assays enable protease monitoring in a high-throughput compatible microtiter plate format, but the methods often rely on either molecular labeling or synthetic protease targets that only mimic the hydrolysis site of the true target proteins. Here, we present a homogenous, label-free, and time-resolved luminescence utilizing the protein-probe method to assay proteases with native and denatured substrates at nanomolar sensitivity. The developed protein-probe method is not restricted to any single protein or protein target class, enabling digestion and substrate fragmentation studies with the natural unmodified substrate proteins. The versatility of the assay for studying protease targets was shown by monitoring the digestion of a substrate panel with different proteases. These results indicate that the protein-probe method not only monitors the protease activity and inhibition, but also studies the substrate specificity of individual proteases.     

Genesis of Active Pt/CeO2 Catalyst for Dry Reforming of Methane by Reduction and Aggregation of Isolated Platinum Atoms into Clusters

Atomically dispersed metal catalysts offer the advantages of efficient metal utilization and high selectivities for reactions of technological importance. Such catalysts have been suggested to be strong candidates for dry reforming of methane (DRM), offering prospects of high selectivity for synthesis gas without coke formation, which requires ensembles of metal sites and is a challenge to overcome in DRM catalysis. However, investigations of the structures of isolated metal sites on metal oxide supports under DRM conditions are lacking, and the catalytically active sites remain undetermined. Data characterizing the DRM reaction-driven structural evolution of a cerium oxide-supported catalyst, initially incorporating atomically dispersed platinum, and the corresponding changes in catalyst performance are reported. X-ray absorption and infrared spectra show that the reduction and agglomeration of isolated cationic platinum atoms to form small platinum clusters/nanoparticles are necessary for DRM activity. Density functional theory calculations of the energy barriers for methane dissociation on atomically dispersed platinum and on platinum clusters support these observations. The results emphasize the need for in-operando experiments to assess the active sites in such catalysts. The inferences about the catalytically active species are suggested to pertain to a broad class of catalytic conversions involving the rate-limiting dissociation of light alkanes.     

Effect of deposition temperature on structural, surface, optical and magnetic properties of pulsed laser deposited Al-doped CdO thin films

The objective of this work is to study the influence of deposition temperature on structural, surface, optical and magnetic properties of the Al doped CdO thin films prepared by pulsed laser deposition (PLD) technique. KrF excimer laser (λ = 248 nm, τ_l = 20 ns, ν = 10 Hz, ?_l = 2.5 J/cm²) was employed for the deposition of thin films. It is observed by XRD results that films grown at room temperature and 100 °C show preferential growth along (1 1 1) and (2 0 0) directions while high temperatures (200-400 °C) lead to preferential growth along the (2 0 0) direction only. The optical constants (n, k, α, and optical band gap energy) of films measured by spectroscopic ellipsometry show strong dependence upon deposition temperature. M-H loop of films, measured by vibrating sample magnetometer, deposited at 25 °C and 100 °C show paramagnetic nature while films deposited at temperatures (200-400 °C) exhibit ferromagnetic character. Scanning electron micrographs show degraded elongated grains at lower deposition temperatures, while smooth and compact surface is observed for films deposited at higher deposition temperatures.     

Harnessing the Extracellular Electron Transfer Capability of Geobacter sulfurreducens for Ambient Synthesis of Stable Bifunctional Single-Atom Electrocatalyst for Water Splitting

Single-atom metal (SA-M) catalysts with high dispersion of active metal sites allow maximum atomic utilization. Conventional synthesis of SA-M catalysts involves high-temperature treatments, leading to low yield with a random distribution of atoms. Herein, a nature-based facile method to synthesize SA-M catalysts (M = Fe, Ir, Pt, Ru, Cu, or Pd) in a single step at ambient temperature, using the extracellular electron transfer capability of Geobacter sulfurreducens (GS), is presented. Interestingly, the SA-M is coordinated to three nitrogen atoms adopting an MN₃ on the surface of GS. Dry samples of SA-Ir@GS without further heat treatment show exceptionally high activity for oxygen evolution reaction when compared to benchmark IrO₂ catalyst and comparable hydrogen evolution reaction activity to commercial 10 wt% Pt/C. The SA-Ir@GS exhibits the best water-splitting performance compared to other SA-M@GS, showing a low applied potential of 1.65 V to achieve 10 mA cm⁻² in 1.0 M KOH with cycling over 5 h. The density functional calculations reveal that the large adsorption energy of H₂O and moderate adsorption energies of reactants and reaction intermediates for SA-Ir@GS favorably improve its activity. This synthesis method at room temperature provides a versatile platform for the preparation of SA-M catalysts for various applications by merely altering the metal precursors.     

Investigating the turbulent flow characteristics in an open channel with staggered vegetation patches

In the present study, flow around circular and staggered vegetation patches was investigated numerically. For turbulence modelling, the Reynolds‐averaged Navier–Stokes technique and Reynolds stress model were adopted. The numerical model was validated with the experimental data using varying vegetation density and flow velocities. The simulated results of mean stream‐wise velocities were in close agreement with the experimental results. The results show that the mean stream‐wise velocity in the downstream regions of vegetation patches were reduced, whereas the velocity in the free stream regions were increased. The influence of neighbouring and staggered vegetation patches on the flow was observed. The vegetation patches with larger nondimensional flow blockage (aD = 2.3, where a is the frontal area per volume of patches, and D is the diameter of vegetation patches) offered more turbulence when compared to the patches with a smaller flow blockage (aD = 1.2). Larger turbulence in the form of kinetic energy and turbulent intensity was recorded within the vegetation as well as the regions directly behind the patches. Negative Reynolds stresses were observed at the top of submerged vegetation. The turbulence characteristics peaked at the top of vegetation, that is, z/h = 1.0 (where z is the flow depth, and h is the vegetation height), which may be migrated vertically as the frontal area of the vegetation patch is increased. This high frontal area also increased stream‐wise velocity above the vegetation, leading to an increased variation in turbulence around the vegetation canopy.     

Design of Simulation System for LTE-U Using 5 GHz Band in MATLAB

Wireless Personal Communications - Unused spectrum is being a limited commodity for the telecom industry. However, up to 500 MHz of unlicensed spectrum in 5 GHz band is in use for...