Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

Effects of preparation conditions on the surface modification and performance of polyethersulfone ultrafiltration membranes

The impact that some membrane preparation steps had on ultrafiltration (UF) membrane characteristics and performance was studied. Polyethersulfone (PES) was employed as base polymer, while N‐methyl pyrrolidone (NMP) was used as a solvent, and polyvinylpyrrolidone (PVP) was used as a nonsolvent pore‐forming additive. The manufacturing variables studied were solvent evaporation time and membrane surface modification, using a fluorine‐based copolymer referred to as surface‐modifying macromolecule (SMM). The flat sheet membranes, prepared via phase inversion, were characterized using solute transport data, X‐ray photoelectron spectroscopy (XPS), and contact angle measurements. Membrane performance was evaluated via filtration test protocol that included a 6‐day filtration of concentrated river water. The flux reduction with time was modeled using single and dual mechanisms of fouling. The pore blockage/cake filtration model described better the behavior of the permeation rate along the experiments. Increasing the solvent evaporation time decreased the size of the pores and the permeation rate. However, it did not significantly affect the removal of the organic compounds naturally present in the river water used as feed. XPS and contact angle measurements proved that the short evaporation periods did not allow enough SMM migration to the surface to provoke a significant effect on the membrane performance. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci, 2006     

Flexible Piezoelectric Touch Sensor by Alignment of Lead‐Free Alkaline Niobate Microcubes in PDMS

A highly sensitive, lead‐free, and flexible piezoelectric touch sensor is reported based on composite films of alkaline niobate K_0.485Na_0.485Li_0.03NbO₃ (KNLN) powders aligned in a polydimethylsiloxane (PDMS) matrix. KNLN powder is fabricated by solid‐state sintering and consists of microcubes. The particles are dispersed in uncured PDMS and oriented by application of an oscillating dielectrophoretic alignment field. The dielectric constant of the composite film is almost independent of the microstructure, while upon alignment the piezoelectric charge coefficient increases more than tenfold up to 17 pC N^?1. A quantitative analysis shows that the origin is a reduction of the interparticle distance to under 1.0 µm in the aligned bicontinuous KNLN chains. The temperature stable piezoelectric voltage coefficient exhibits a maximum value of 220 mV m N^?1, at a volume fraction of only 10%. This state‐of‐the‐art value outperforms bulk piezoelectric ceramics and composites with randomly dispersed particles, and is comparable to the values reported for the piezoelectric polymers polyvinylidenefluoride and its random copolymer with trifluoroethylene. Optimized composite films are incorporated in flexible piezoelectric touch sensors. The high sensitivity is analyzed and discussed. As the fabrication technology is straightforward and easy to implement, applications are foreseen in flexible electronics such as wireless sensor networks and biodiagnostics.     

The Endolysosomal System: The Acid Test for SARS-CoV-2

This review aims to describe and discuss the different functions of the endolysosomal system, from homeostasis to its vital role during viral infections. We will initially describe endolysosomal system’s main functions, presenting recent data on how its compartments are essential for host defense to explore later how SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) and other coronaviruses subvert these organelles for their benefit. It is clear that to succeed, pathogens’ evolution favored the establishment of ways to avoid, escape, or manipulate lysosomal function. The unavoidable coexistence with such an unfriendly milieu imposed on viruses the establishment of a vast array of strategies to make the most out of the invaded cell’s machinery to produce new viruses and maneuvers to escape the host’s defense system.     

Sugarcane bagasse cellulose/HDPE composites obtained by extrusion

Natural fibers used in this study were both pre-treated and modified residues from sugarcane bagasse. Polymer of high density polyethylene (HDPE) was employed as matrix in to composites, which were produced by mixing high density polyethylene with cellulose (10%) and Cell/ZrO₂·nH₂O (10%), using an extruder and hydraulic press. Tensile tests showed that the Cell/ZrO₂·nH₂O (10%)/HDPE composites present better tensile strength than cellulose (10%)/HDPE composites. Cellulose agglomerations were responsible for poor adhesion between fiber and matrix in cellulose (10%)/HDPE composites. HDPE/natural fibers composites showed also lower tensile strength in comparison to the polymer. The increase in Young’s modulus is associated to fibers reinforcement. SEM analysis showed that the cellulose fibers insertion in the matrix caused an increase of defects, which were reduced when modified cellulose fibers were used.     

Characterization of peptide and amino acid profile in casein hydrolysates

The nutritional quality of protein hydrolysates has been related in several reports to their di- and tripeptide contents. In the present work different hydrolytic conditions were tested using papain in order to prepare casein hydrolysates with a suitable peptide profile for being used in special diets. The hydrolysates were fractionated by size-exclusion HPLC and the rapid Correct Fraction Area method was used for quantifying the peptides. Among the five hydrolytic conditions studied, three of them gave rise to preparations having nutritionally similar peptide profiles. However, the use of the temperature of 37 degrees C and enzyme:substrate ratio (E:S) of 2% may probably be the most economical condition for industrial production.     

The Topology,in Model Membranes,of the Core Peptide Derived from the T‐Cell Receptor Transmembrane Domain

The T‐cell receptor–CD3 complex (TCR–CD3) serves a critical role in protecting organisms from infectious agents. The TCR is a heterodimer composed of α‐ and β‐chains, which are responsible for antigen recognition. Within the transmembrane domain of the α‐subunit, a region has been identified to be crucial for the assembly and function of the TCR. This region, termed core peptide (CP), consists of nine amino acids (GLRILLLKV), two of which are charged (lysine and arginine) and are crucial for the interaction with CD3. Earlier studies have shown that a synthetic peptide corresponding to the CP sequence can suppress the immune response in animal models of T‐cell‐mediated inflammation, by disrupting proper assembly of the TCR. As a step towards the understanding of the source of the CP activity, we focused on CP in egg phosphatidylcholine/cholesterol (9:1, mol/mol) model membranes and determined its secondary structure, oligomerization state, and orientation with respect to the membrane. To achieve this goal, 15‐residue segments of TCRα, containing the CP, were synthesized and spin‐labeled at different locations with a nitroxide derivative. Electron spin‐echo envelope modulation spectroscopy was used to probe the position and orientation of the peptides within the membrane, and double electron–electron resonance measurements were used to probe its conformation and oligomerization state. We found that the peptide is predominantly helical in a membrane environment and tends to form oligomers (mostly dimers) that are parallel to the membrane plane.     

Impact of Membrane Surface Modification on the Treatment of Surface Water

New polyethersulfone (PES) based membranes for ultrafiltration (UF) were developed by blending a surface-modifying macromolecule (SMM) in the casting solution, in an attempt to minimize the impact of fouling. Fouling was evaluated using concentrated Ottawa River water (CORW), either unfractionated or fractionated via UF. These membranes also included some polyvinylpyrrolidone (PVP), a pore forming additive. A statistical analysis was conducted to evaluate the impact of some variables on the treatment of the surface water. The independent variables included PVP/PES ratio in the casting solution, with and without SMM, and the nature of the feed CORW [low molecular weight (LMW) fraction, unfractionated, high molecular weight (HMW) fraction]. The performance variables studied were total organic carbon (TOC) removal, the foulant accumulation at the membrane surface after filtration, the flux reduction, and the final permeate flux. The most important variable was the feed water. Filtration of LMW had a higher final flux, less fouling, but slightly lower TOC removal. The SMM did not significantly impact the membrane performance. TOC removal was high, compared with results reported in the literature for UF membranes.