Hartvig French (patchwound4)
Zinc (Zn) alloys are promising alternatives to magnesium (Mg)- and iron (Fe)-based alloys because of their moderate corrosion rate and superior biocompatibility. To reduce the mass release of Zn2+ and improve the biocompatibility of Zn implants, the biomimetic zwitterionic polymer layer (phosphorylcholine chitosan-PCCs) was immobilized on the plasma-treated Zn1Mg surface. It is the chemical bonds between the -NH2 groups of the PCCs chain and O-C═O (C═O) groups on the plasma-treated Zn1Mg (Zn1Mg-PP) that contributes to the strong bonding strength between the film and the substrate, by which the PCCs (approx. 200 nm thick) layer can bear a 5.93 N normal load. The electrochemical impedance spectroscopy (EIS) results showed that the PCCs layer remarkably increased the resistance against corrosion attack, protecting substrates from over-quick degradation, and the protective effect of the layer with a thickness of 200 nm lasts for about 24 h. The corrosion products of Zn1Mg-PP-PCC in NaCl solution were determined as Zn5(OH)8Cl2·H2O and Zn3(PO4)2. Besides, the bulk Zn1Mg can trigger more aggressive macrophage activity, while the surface of Zn1Mg-PP and Zn1Mg-PP-PCC and their corrosion products (Zn3(PO4)2) tend to promote the differentiation of macrophages into the M2 phenotype, which is beneficial for implant applications.Affordable and automated cloning platforms are essential to many synthetic biology studies. However, the traditional E. coli-based cloning is a major bottleneck as it requires heat shock or electroporation implemented in the robotic workflows. To overcome this problem, we explored bacterial natural transformation for automatic DNA cloning and engineering. Recombinant plasmids are efficiently generated from Gibson or overlap extension PCR (OE-PCR) products by simply adding the DNA into Acinetobacter baylyi ADP1 cultures. No DNA purification, competence induction, or special equipment is required. Up to 10,000 colonies were obtained per microgram of DNA, while the number of false positive colonies was low. We cloned and engineered 21 biosynthetic gene clusters (BGCs) of various types, with length from 1.5 to 19 kb and GC content from 35% to 72%. One of them, a nucleoside BGC, showed antibacterial activity. Furthermore, the method was easily transferred to a low-cost benchtop robot with consistent cloning efficiency. Thus, this automatic natural transformation (ANT) cloning provides an easy, robust, and affordable platform for high throughput DNA engineering.Lanthanides such as cerium(III), europium(III), and gadolinium(III) are widely used for designing fluorescent probes or magnetic resonance imaging contrasting agents for biological systems. The synthesis and study of lanthanide complexes in buffer solutions imitating biological fluids are often complicated because of a lack of data on the lanthanide interactions with buffer solution components. Therefore, Ln(III) [where Ln(III) = La(III), Ce(III), Gd(III), Eu(III)] complexation with a widely used buffer agent, tris(hydroxymethyl)aminomethane (Tris), in aqueous solution is studied using potentiometry, spectrofluorimetry, and 139La NMR spectroscopy. IRAK-1-4 Inhibitor I supplier The stoichiometric composition of complexes is determined using mass spectrometry. The thermodynamic stability constants of Ln(III)-Tris complexes are calculated from potentiometric and spectral data; the difficulties in the study of these systems, reliability, and accuracy of the obtained constants are discussed. The possible structures of free Tris and its complexes with lanthanides(III) are optimized on the density functional theory/PBE0 level; the peculiarities of metal-ligand bonds were studied by Quantum Theory Atoms in Molecules analysis.Single-cell mass spectrometry (MS) remains challenging in the analysis of cells in the native environment due to the severe ion suspension from nonvolatile salts. Synchronous desalting and ionization would be ideal to both ensure the native environ