Creation of bioorthogonal redox systems depending on nicotinamide flucytosine dinucleotide

Many enzymes catalyzing biological redox chemistry depend on the omnipresent cofactor, nicotinamide adenine dinucleotide (NAD). NAD is also involved in various nonredox processes. It remains challenging to disconnect one particular NAD-dependent reaction from all others. Here we present a bioorthogonal system that catalyzes the oxidative decarboxylation of l-malate with a dedicated abiotic cofactor, nicotinamide flucytosine dinucleotide (NFCD). By screening the multisite saturated mutagenesis libraries of the NAD-dependent malic enzyme (ME), we identified the mutant ME-L310R/Q401C, which showed excellent activity with NFCD, yet marginal activity with NAD. We found that another synthetic cofactor, nicotinamide cytosine dinucleotide (NCD), also displayed similar activity with the ME mutants. Inspired by these observations, we mutated d-lactate dehydrogenase (DLDH) and malate dehydrogenase (MDH) to DLDH-V152R and MDH-L6R, respectively, and both mutants showed fully active with NFCD. When coupled with DLDH-V152R, ME-L310R/Q401C required only a catalytic amount of NFCD to convert l-malate. Our results opened the window to engineer bioorthogonal redox systems for a wide variety of applications in systems biology and synthetic biology.     

An SPR biosensor for the detection of microcystins in drinking water

A surface plasmon resonance (SPR) biosensor for the detection of microcystins (MCs) in drinking water has been developed. Several assay formats have been evaluated. The selected format is based on a competitive inhibition assay, in which microcystin-LR (MCLR) has been covalently immobilized onto the surface of an SPR chip functionalized with a self-assembled monolayer. The influence of several factors affecting sensor performance, such as the nature and concentration of the antibody, the composition of the carrier buffer, and the blocking and regeneration solutions, has been evaluated. The optimized SPR biosensor provides an IC₅₀ 0.67 ± 0.09 μg L⁻¹, a detection limit of 73 ± 8 ng L⁻¹, and a dynamic range from 0.2 to 2.0 μg L⁻¹ for MCLR. Cross-reactivity to other related MCs, such as microcystin-RR (88%) and microcystin-YR (94%), has also been measured. The SPR biosensor can perform four simultaneous determinations in 60 min, and each SPR chip can be reused for at least 40 assay–regeneration cycles without significant binding capacity loss. The biosensor has been successfully applied to the direct analysis of MCLR in drinking water samples, below the provisional guideline value of 1 μg L⁻¹ established by the World Health Organization for drinking water.     

Thermal characterization and compatibility studies of perovskite-type Ba0.5Sr0.5Co0.8Fe0.2O3−δ (BSCF) oxide with Cr2O3 at high temperature

The chemical compatibility of perovskite-type Ba_0.5Sr_0.5Co_0.8Fe_0.2O_3?δ (BSCF) oxides with Cr₂O₃ has been examined between room temperature and 1,100 °C. Differential thermal analysis and thermogravimetric analysis were used to analyze the thermal behavior of BSCF–Cr₂O₃ binary mixtures in all composition ranges (0–100 mass% BSCF). The reaction products were identified by X-ray analysis after heating at 700–1,100 °C. As we expected, it was found that perovskite-type BSCF oxide had a poor chemical compatibility with the Cr₂O₃ oxide. In particular, the decomposition process of the BSCF–Cr₂O₃ binary mixture is quite complex and it starts at about 700–750 °C. The mixtures of BSCF and Cr₂O₃ oxides reacted forming mixed complex oxides based on (Ba/Sr)FeO₃, (Co/Fe)CrO₄, and (Ba/Sr)CrO₄ mixtures.     

Structure-tuned membrane active Ir-complexed oligoarginine overcomes cancer cell drug resistance and triggers immune responses in mice

The development of chemotherapy, an important cancer treatment modality, is hindered by the frequently found drug-resistance phenomenon. Meanwhile, researchers have been enthused lately by the synergistic use of chemotherapy with emerging immunotherapeutic treatments. In an effort to address both of the two unmet needs, reported herein is a study on a series of membrane active iridium(iii) complexed oligoarginine peptides with a new cell death mechanism capable of overcoming drug resistance as well as stimulating immunological responses. A systematic structure–activity relationship study elucidated the interdependent effects of three structural factors, i.e., hydrophobicity, topology and cationicity, on the regulation of the cytotoxicity of the Ir(iii)-oligoarginine peptides. With the most prominent toxicities, Ir-complexed octaarginines (R8) were found to display a progressive oncotic cell death featuring cell membrane-penetration and eruptive cytoplasmic content release. Consequently, this membrane-centric death mechanism showed promising potential in overcoming multiple chemical drug-resistance of cancer cells. More interestingly, the eruptive mode of cell death proved to be immunogenic by stimulating the dendritic cell maturation and inflammatory factor accumulation in mice tumours. Taking these mechanisms together, this work demonstrates that membrane active compounds may become the next generation chemotherapeutics because of their combined advantages.

Structure optimized Ir-complexed cyclic octa-arginine shows a potential of “one-drug two-effects” for cancer treatment.     

Friedel-Crafts alkylation of indoles with nitroalkenes catalyzed by Cu(Ⅱ)-imine complex

A series of new ligands L₁-L₇ were readily prepared in one step.Friedel-Crafts alkylation of indoles with nitroalkenes catalyzed by a novel Cu（Ⅱ）-L complex has been developed.The remarkable advantages of this reaction are mild reaction conditions,simple workup procedure,high yields of products and the use of ethanol as a green solvent.     

Synthesis and photophysical properties of α,ω-bis(terpyridine)oligothiophenes

Four novel fully π-conjugated α,ω-bis(terpyridine)oligothiophenes characterized by NMR, IR, and HR-mass spectroscopy are presented and their electronic absorption/emission and redox properties are described based on both experiments and theoretical calculations. These compounds can be potentially utilized as building blocks for preparation of conjugated metallo-supramolecular polymers or dynamers and related functional materials.     

Modeling mass transfer of CO2 in brine at high pressures by chemical potential gradient

To investigate long-term CO2 behavior in geological formations and quantification of possible CO2 leaks, it is crucial to investigate the potential mobility of CO2 dissolved in brines over a wide range of spatial and temporal scales and density distributions in geological media. In this work, the mass transfer of aqueous CO 2 in brines has been investigated by means of a chemical potential gradient model based on non-equilibrium thermodynamics in which the statistical associating fluid theory equation of state was used to calculate the fugacity coefficient of CO2 in brine. The investigation shows that the interfacial concentration of aqueous CO2 and the corresponding density both increase with increasing pressure and decreasing temperature; the effective diffusion coefficients decrease initially and then increase with increasing pressure; and the density of the CO2 -disolved brines increases with decreasing CO2 pressure in the CO2 dissolution process. The aqueous CO2 concentration profiles obtained by the chemical potential gradient model are considerably different from those obtained by the concentration gradient model, which shows the importance of considering non-ideality, especially when the pressure is high.     

Characterization of major flavonoids, triterpenoid, dipeptide and their metabolites in rat urine after oral administration of Radix Astragali decoction

To characterize of the constituents in rat urine after oral administration of Radix Astragali decoction, a HPLC-DAD-MS/MS technique had been developed. Urine collected from 0 to 24 h, after administration, was purified using a C18 solid-phase extraction cartridge, and then detected by an on-line MS/MS detector. By comparing the retention times and MS/MS data with those obtained from authentic compounds and the published data, a total of 11 compounds including six flavonoids, one dipeptide, one triterpenoid and three of their metabolites in urine were identified. Compounds daidzein, genistein, quercetin, L-asparamide-D-phenylalanine, 4,2′,4′-trihydroxychalcone, Astragaloside-IV, daidzein sulfate, and kaempferol sulfate were for the first time detected by HPLC-MS/MS technique in urine. The present research results success-fully narrowed the range of effective constituents to be found in Astragalus membranaceus and would be useful for the following action mechanism research of this traditional Chinese medicine in treating various diseases.     

Accessing the long-lived emissive 3IL triplet excited states of coumarin fluorophores by direct cyclometallation and its application for oxygen sensing and upconversion

We studied four cyclometallated Pt(II) complexes, in which the thiazo-coumarin ligands (Pt-2, Pt-3 and Pt-4) or the phenylthiazo ligand (Pt-1) were directly cycloplatinated. Pt-2 shows intense absorption in visible region but other complexes show blue-shifted absorption. Room temperature phosphorescence was observed for all the complexes, and the emission wavelength is dependent on the size of the π-conjugation, not the intramolecular charge transfer (ICT) feature of the C^N ligands. Pt-2 shows longer phosphorescence lifetime (τ = 20.3 μs) than other complexes (below 2.0 μs). Furthermore, Pt-2 shows phosphorescence quantum yield Φ = 0.37, whereas Pt-3 and Pt-4 show much smaller Φ values (0.03 and 0.01, respectively). DFT/TDDFT calculations indicate (3)IL triplet excited states for the complexes. The complexes were used as for luminescence O(2) sensing and triplet-triplet-annihilation (TTA) based upconversion. Stern-Volmer quenching constant K(SV) = 0.026 Torr(-1) was observed for Pt-2, ca. 89-fold of that of Pt-3. TTA upconversion is achieved with Pt-2 (λ(em) = 400 nm with λ(ex) = 473 nm, anti-Stokes shift is 0.47 eV, excitation power density is at 70 mW cm(-2)). The upconversion quantum yield with Pt-2 as triplet sensitizer is up to 15.4%. The TTET efficiency (K(SV) = 1.33 × 10(5) M(-1), k(q) = 6.57 × 10(9) M(-1) s(-1). DPA as quencher) of Pt-2 is 34-fold of the model complex [Ru(dmb)(3)][PF(6)](2). Our results show that the (3)IL state can be readily accessed by direct cyclometallation of organic fluorophores and this approach will be useful for preparation and applications of transition metal complexes that show intense absorption in visible region and the long-lived emissive (3)IL excited states.