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     

Envelopes with microplastics generated from recycled plastic bags for crude oil sorption

Oil spill accidents in marine environments and the lack of disposal of post-consumer plastic are environmental problems worldwide. This study presents a sustainable alternative for both issues through envelopes filled with microplastics (MPs) from recycled bags for the sorption of spilled crude oil. Through particle size analysis by three different sieves (4, 9, and 20 mesh), different MP sizes were characterized by scanning electron microscope (SEM), Brunauer–Emmett–Teller (BET), density, contact angle (CA), and Fourier-transform infrared spectroscopy (FTIR). According to their sizes, the MPs were distributed in envelopes and submitted to crude oil sorption capacity and efficiency evaluation. Three MP particle sizes were obtained (from the largest to the smallest, according to the sieve mesh, MP4, MP9, and MP20). SEM images of samples exhibited irregular and porous surfaces, and MP4 had the smallest pore size (8.6 μm). BET showed that MP4 had the highest surface area (0.074 m²/g). The CA > 90° exposed that all samples were hydrophobic. FTIR spectrum demonstrated that the samples from the recycled bags were made of high density polyethylene (HDPE). The MP4 envelopes also had the best crude oil sorption results in capacity and efficiency (1.73 g/g and 68%, respectively), being a promising recycled sorbent in crude oil spillage applications.     

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.     

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.     

Dietary fatty acids early in life affect lipid metabolism and adiposity in young rats

The purpose of this study was to evaluate the effects of four isoenergetic diets of differing fat composition on blood lipid profile and adiposity in young rats. Diets containing different lipid sources—partially hydrogenated vegetable oil (PHVO), palm oil (PO), canola oil (CO), and soy oil (SO)—were fed to lactating rats during the 21 days of lactation, and then fed to young males following weaning until the 45th, day of life. In vivo lipogenesis rate (LR), lipid content (LC), relative level of FA, and the activity of lipoprotein lipase (LPL) enzyme were measured in epididymal adipose tissue (EPI). Fasting blood lipoproteins and LC in the carcass were also appraised. Body weight of PO and PHVO groups was significantly higher than CO and SO groups from day 14 of lactation to day 45, despite the lower food intake in the PHVO group. PO and PHVO groups presented higher LR and LC in EPI than SO and CO groups. Carcass fat content was significantly higher in PHVO and PO groups than in CO and SO groups. The LPL activity in EPI was unaffected by dietary lipids. PHVO group had increased total cholesterol and TAG concentrations in comparison with the PO group, and significantly lower HDL level compared with the other groups. These results show that the kind of FA in the dietary lipid offered early in life can affect lipid metabolism and adiposity.