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In this study, we report an easy approach for the production of aqueous dispersions of C60 fullerene with good stability. Maleic acid copolymers, poly(styrene-alt-maleic acid) (SM), poly(N-vinyl-2-pyrrolidone-alt-maleic acid) (VM) and poly(ethylene-alt-maleic acid) (EM) were used to stabilize C60 fullerene molecules in an aqueous environment by forming non-covalent complexes. Polymer conjugates were prepared by mixing a solution of fullerene in N-methylpyrrolidone (NMP) with an aqueous solution of the copolymer, followed by exhaustive dialysis against water. The molar ratios of maleic acid residues in the copolymer and C60 were 5/1 for SM and VM and 10/1 for EM. The volume ratio of NMP and water used was 1:1.2–1.6. Water-soluble complexes (composites) dried lyophilically retained solubility in NMP and water but were practically insoluble in non-polar solvents. The optical and physical properties of the preparations were characterized by UV-Vis spectroscopy, FTIR, DLS, TGA and XPS. The average diameter of the composites in water was 120–200 nm, and the ξ-potential ranged from −16 to −20 mV. The bactericidal properties of the obtained nanostructures were studied. Toxic reagents and time-consuming procedures were not used in the preparation of water-soluble C60 nanocomposites stabilized by the proposed copolymers.

A simple one-pot method is proposed for obtaining the colloidal nanohybrid structures of silver (gold) and zinc oxide as well as nanostructures doped with zinc ions. The copolymers of maleic acid were used for the stabilization of nanoheterostructures. To characterize the preparation, UV–Vis spectroscopy, TEM, FTIR, XPS, and XRD were used. The bactericidal properties of the nanoheterostructures were studied in relation to the fungus C. albicans and the bacteria E. coli and S. aureus, used in planktonic form. In general, the samples containing nanosilver were the most active, and the preparations containing gold nanoparticles were the least active. The minimum inhibitory concentrations (MICs) of the Ag/ZnO samples, based on all copolymers, were in the ranges of 1.4–1.7 μg/mL for C. albicans, 2.9–6.8 μg/mL for E. coli, and 23–27 μg/mL for S. aureus; the MIC values of Au/ZnO samples were 472 μg/mL for S. aureus and 945 μg/mL for C. albicans and E. coli. The additional introduction of zinc cations into heterodimers had practically no effect on the antimicrobial properties of the composites. For all prepared composites and all tested microorganisms, the fractional inhibitory concentration indexes were in the range of 0.5–2.2, which indicates a close-to-additive contribution of the bioactive components in the samples used in the bactericidal process.

This study aimed to enhance homology-directed repair (HDR) efficiency in CRISPR/Cas-mediated genome editing by targeting three key factors regulating the balance between HDR and non-homologous end joining (NHEJ): MAD2L2, SCAI, and Ligase IV. In order to achieve this, a cellular model using mutated eGFP was designed to monitor HDR events. Results showed that MAD2L2 knockdown and SCR7 treatment significantly improved HDR efficiency during Cas9-mediated HDR repair of the mutated eGFP gene in the HEK293T cell line. Fusion protein Cas9-SCAI did not improve HDR. This study is the first to demonstrate that MAD2L2 knockdown during CRISPR-mediated gene editing in HEK293T cells can increase precise correction by up to 10.2 times. The study also confirmed a moderate but consistent effect of SCR7, an inhibitor of Ligase IV, which increased HDR by 1.7 times. These findings provide valuable insights into improving HDR-based genome editing efficiency.

In this study, we report an easy approach for the production of aqueous dispersions of C60 fullerene with good stability. Maleic acid copolymers, poly(styrene-alt-maleic acid) (SM), poly(N-vinyl-2-pyrrolidone-alt-maleic acid) (VM) and poly(ethylene-alt-maleic acid) (EM) were used to stabilize C60 fullerene molecules in an aqueous environment by forming non-covalent complexes. Polymer conjugates were prepared by mixing a solution of fullerene in N-methylpyrrolidone (NMP) with an aqueous solution of the copolymer, followed by exhaustive dialysis against water. The molar ratios of maleic acid residues in the copolymer and C60 were 5/1 for SM and VM and 10/1 for EM. The volume ratio of NMP and water used was 1:1.2–1.6. Water-soluble complexes (composites) dried lyophilically retained solubility in NMP and water but were practically insoluble in non-polar solvents. The optical and physical properties of the preparations were characterized by UV-Vis spectroscopy, FTIR, DLS, TGA and XPS. The average diameter of the composites in water was 120–200 nm, and the ξ-potential ranged from −16 to −20 mV. The bactericidal properties of the obtained nanostructures were studied. Toxic reagents and time-consuming procedures were not used in the preparation of water-soluble C60 nanocomposites stabilized by the proposed copolymers. [ABSTRACT FROM AUTHOR]

In vitro activity of dioxidin against pathogens of nosocomial infections and its cytotoxicity were estimated. The study involved 300 isolates from patients with nosocomial infections. The MICs of dioxidin were determined with the method of serial dilutions in broth. The dioxidin cytotoxicity was investigated with the MTT assay to assign the cell culture viability. In concentrations of 2 to 1024 mcg/ml dioxidin was active against 279/300 (93%) strains. The drug inhibited the growth of all the gramnegative isolates. The highest activity was observed against Enterobacteriaceae vs. nonfermenting gramnegative bacteria: the median, minimum and maximum MICs of dioxidin were 12 (4-32) and 32 (16-64) mcg/ml respectively. The dioxidin activity against gramnegative bacteria and fungi was lower. The MIC of dioxidin for 7/70 (10%) staphylococcal isolates, 9/28 (32%) enterococcal isolates and all the Candida isolates was > 1024 mcg/ml. The IC50 of dioxidin was 2.4+0.3 mM (low cytotoxicity). The results showed that the use of dioxidin as an antimicrobial for local application was advisable in the treatment of gramnegative bacterial infections provided adequate tissue concentrations were attained.

Recent advances in machine learning techniques have led to development of a number of protein design and engineering approaches. One of them, ProteinMPNN, predicts an amino acid sequence that would fold and match user‐defined backbone structure. Its performance was previously tested for proteins composed of standard amino acids, as well as for peptide‐ and protein‐binding proteins. In this short report, we test whether ProteinMPNN can be used to reengineer a non‐proteinaceous ligand‐binding protein, flavin‐based fluorescent protein CagFbFP. We fixed the native backbone conformation and the identity of 20 amino acids interacting with the chromophore (flavin mononucleotide, FMN) while letting ProteinMPNN predict the rest of the sequence. The software package suggested replacing 36–48 out of the remaining 86 amino acids so that the resulting sequences are 55%–66% identical to the original one. The three designs that we tested experimentally displayed different expression levels, yet all were able to bind FMN and displayed fluorescence, thermal stability, and other properties similar to those of CagFbFP. Our results demonstrate that ProteinMPNN can be used to generate diverging unnatural variants of fluorescent proteins, and, more generally, to reengineer proteins without losing their ligand‐binding capabilities. [ABSTRACT FROM AUTHOR]

This research is focused on the adsorption modification of detonation nanodiamond surfaces with antibiotics for their further use as smart materials for cardiovascular surgery purposes, namely as bioprostheses modifiers. Tritium-labeled amikacin and levofloxacin were used as tracers for the adsorption process control. We found that nanodiamonds form adsorption complexes with levofloxacin via physical adsorption, while in the case of amikacin, electrostatic attraction contributes to the formation of more stable complexes, even in the presence of electrolytes and desorbing agents (models of biological fluids). Antimicrobial characterization of nanodiamond–levofloxacin and nanodiamond–amikacin complexes indicates a reduction in the dose of antibiotics that is used as an antimicrobial agent. Therefore, the use of biomaterial based on DND complexes with antibiotics as the basis of bioprostheses will allow one either to avoid or significantly reduce the duration and intensity of antibiotics use in the postoperative period, which is critically important from the viewpoint of the development of antibiotic resistance in pathogens.

Development of genome editing methods created new opportunities for the development of etiology-based therapies of hereditary diseases. Here, we demonstrate that CRISPR/Cas9 can correct p.F508del mutation in the CFTR gene in the CFTE29o- cells and induced pluripotent stem cells (iPSCs) derived from patients with cystic fibrosis (CF). We used several combinations of Cas9, sgRNA and ssODN and measured editing efficiency in the endogenous CFTR gene and in the co-transfected plasmid containing the CFTR locus with the p.F508del mutation. The non-homologous end joining (NHEJ) frequency in the CFTR gene in the CFTE29o- cells varied from 1.25% to 2.54% of alleles. The best homology-directed repair (HDR) frequency in the endogenous CFTR locus was 1.42% of alleles. In iPSCs, the NHEJ frequency in the CFTR gene varied from 5.5% to 12.13% of alleles. The best HDR efficacy was 2.38% of alleles. Our results show that p.F508del mutation editing using CRISPR/Cas9 in CF patient-derived iPSCs is a relatively rare event and subsequent cell selection and cultivation should be carried out.