An easy tool to monitor the elemental steps of in vitro translation via gel electrophoresis of fluorescently labeled small peptides.

Several methods are available to visualize and assess the kinetics and efficiency of elemental steps of protein biosynthesis. However, each of these methods has its own limitations. Here, we present a novel, simple and convenient tool for monitoring stepwise in vitro translation initiated by BODIPY-Met-tRNA. Synthesis and release of very short, 1-7 amino acids, BODIPY-labeled peptides, can be monitored using urea-polyacrylamide gel electrophoresis. Very short BODIPY-labeled oligopeptides might be resolved this way, in contrast to widely used Tris-tricine gel electrophoresis, which is suitable to separate peptides larger than 1 kDa. The method described in this manuscript allows one to monitor the steps of translation initiation, peptide transfer, translocation, and termination as well as their inhibition at an unprecedented single amino acid resolution.

Antioxidant Activity of New Copolymer Conjugates of Methoxyoligo(Ethylene Glycol)Methacrylate and Betulin Methacrylate with Cerium Oxide Nanoparticles In Vitro.

The synthesis of two new copolymer conjugates of methoxyoligo(ethylene glycol)methacrylate MPEGMA and betulin methacrylate BM was developed via RAFT polymerization. The molar content of BM units was equal to 9-10 and 13-16 mol%, respectively (HPLC, 1H and 13C NMR); molar weights were equal to 75000-115000. CeO2 NPs as a component of the hybrid material were synthesized for the preparation of the composition with copolymer conjugates of MPEGMA and BM. We showed a significant increase in G6PDH and GR activities by 21-51% and 9-132%, respectively, which was due to the increase in NADPH concentration under the action of copolymers in vitro. The actions of copolymers and CeO2 NPs combination were stronger than those of the individual components: the SOD activity increased by more than 30%, the catalase activity increased dose-dependently from 13 to 45%, and the GR activity increased to 49%. The maximum increase in enzyme activity was observed for the G6PDH from 54% to 151%. The MDA level dose-dependently increased by 3-15% under the action of copolymers compared with the control, and dose-dependently decreased by 3-12% in samples containing CeO2 NPs only. CeO2 NP-copolymer compositions can be used for the design of new biomimetic medical products with controlled antioxidant properties.

Concentration effects in the base-catalyzed hydrolysis of oligo(ethylene glycol)- and amine-containing methacrylic monomers.

Concentration effects in the base-catalyzed hydrolysis of water-soluble methacrylates (3-(N,N-dimethylaminoethyl) methacrylate (DMAEMA), 2-hydroxyethyl methacrylate (HEMA) and oligo(ethylene glycol) methacrylates (OEGMAs)) have been studied. These monomers are rapidly hydrolyzed in the presence of bases at the room temperature in dilute aqueous solutions, but the reaction rate decreases sharply in highly concentrated solutions. A clear correlation was found between a form of the viscosity isotherm for DMAEMA solutions and the concentration dependence of the autocatalytic hydrolysis rate which indicates the connection of process kinetics with the structure of solutions. These data should be considered when carrying out homo- and copolymerization of the previously mentioned monomers in aqueous solutions.

Amide-Containing Bottlebrushes via Continuous-Flow Photoiniferter Reversible Addition-Fragmentation Chain Transfer Polymerization: Micellization Behavior.

Herein, a series of ternary amphiphilic amide-containing bottlebrushes were synthesized by photoiniferter (PI-RAFT) polymerization of macromonomers in continuous-flow mode using trithiocarbonate as a chain transfer agent. Visible light-mediated polymerization of macromonomers under mild conditions enabled the preparation of thermoresponsive copolymers with low dispersity and high yields in a very short time, which is not typical for the classical reversible addition-fragmentation chain transfer process. Methoxy oligo(ethylene glycol) methacrylate and alkoxy(C12-C14) oligo(ethylene glycol) methacrylate were used as the basic monomers providing amphiphilic and thermoresponsive properties. The study investigated how modifying comonomers, acrylamide (AAm), methacrylamide (MAAm), and N-methylacrylamide (-MeAAm) affect the features of bottlebrush micelle formation, their critical micelle concentration, and loading capacity for pyrene, a hydrophobic drug model. The results showed that the process is scalable and can produce tens of grams of pure copolymer per day. The unmodified copolymer formed unimolecular micelles at temperatures below the LCST in aqueous solutions, as revealed by DLS and SLS data. The incorporation of AAm, MAAm, and N-MeAAm units resulted in an increase in micelle aggregation numbers. The resulting bottlebrushes formed uni- or bimolecular micelles at extremely low concentrations. These micelles possess a high capacity for loading pyrene, making them a promising choice for targeted drug delivery.

Zinc Oxide Nanoparticles Protected with Terpenoids as a Substance in Redox Imbalance Normalization in Burns.

Preliminary protection of zinc oxide nanoparticles (ZnO NPs) with terpenoids such as betulin, its derivatives, and essential oils components has been proposed to produce gel-like oleophilic and hydrophilic formulations. We studied the properties of gel-like dispersions of ZnO NPs with immobilized terpenoids and their effects on the activity of LDH, GR, G6PDH, restoration of redox balance of co-enzyme pairs NAD+/NADH and NADP+/NADPH, as well as the activity of SOD, catalase, AlDH in erythrocytes in the treatment of burns in rats. Hysteresis loops on the rheograms of studied dispersions characterize their thixotropic properties. ZnO NPs with betulin diphosphate in the water-ethanol medium lead to a 20-fold increase in the hydrodynamic radius at pH 7.3 compared to the initial ZnO NPs, and facilitate the formation of Zn2+ ions and their penetration into the viable epidermis, unlike oleophilic dispersions. All dispersions reduce the healing time by one and a half times compared with the untreated control group, increase the activity of LDH, GR, G6PDH, SOD, catalase, AlDH, and contribute to the normalization of coenzyme balance. Normalization of the redox balance and wound state was more effective using hydrophilic dispersions due to Zn2 + penetration.

Random and Diblock Thermoresponsive Oligo(ethylene glycol)-Based Copolymers Synthesized via Photo-Induced RAFT Polymerization.

Amphiphilic random and diblock thermoresponsive oligo(ethylene glycol)-based (co)polymers were synthesized via photoiniferter polymerization under visible light using trithiocarbonate as a chain transfer agent. The effect of solvent, light intensity and wavelength on the rate of the process was investigated. It was shown that blue and green LED light could initiate RAFT polymerization of macromonomers without an exogenous initiator at room temperature, giving bottlebrush polymers with low dispersity at sufficiently high conversions achieved in 1-2 h. The pseudo-living mechanism of polymerization and high chain-end fidelity were confirmed by successful chain extension. Thermoresponsive properties of the copolymers in aqueous solutions were studied via turbidimetry and laser light scattering. Random copolymers of methoxy- and alkoxy oligo(ethylene glycol) methacrylates of a specified length formed unimolecular micelles in water with a hydrophobic core consisting of a polymer backbone and alkyl groups and a hydrophilic oligo(ethylene glycol) shell. In contrast, the diblock copolymer formed huge multimolecular micelles.

The New Pharmaceutical Compositions of Zinc Oxide Nanoparticles and Triterpenoids for the Burn Treatment.

We studied oleogels containing zinc oxide nanoparticles (ZnO NPs) and lupane triterpenoids in sunflower oil for the treatment of burns. The modification of ZnO was carried out by treatment with alcohol solutions of betulin, betulonic acid, betulin diacetate and betulin diphosphate. The properties of modified ZnO NPs were studied by powder XRD (average sizes of 10-20 nm), FTIR (νZnO 450 cm-1), UV-vis (345-360 nm), and blue-violet emission (380-420 nm). The identification and assay of modified ZnO NPs and triterpenoids were estimated. The treatment by oleogels of deep II-degree burns was studied on rats using histological studies, Doppler flowmetry and evaluation of enzymes activity and malonic dialdehyde (MDA) level. After the action of oleogels, burn wound area, and the necrosis decreased twice on the 10th day in comparison with the 1st day after burn. The microcirculation index in the near-wound zone by 20-30% improved compared with the group without treatment. Evaluation of the enzyme activity and the MDA level after treatment by oleogels during the course of 10 days showed them returning to normal. The improvement of antioxidant biochemical indexes, as well as wounds' healing, was mainly determined by the influence of zinc oxide nanoparticles.