Design,synthesis, crystal structure and in vitro cytotoxic properties of a novel Schiff base derived from indole and biphenyl

A novel and potentially active dihydroorotate dehydrogenase (DHODH) inhibitor, namely 3‐({(E )‐[(E )‐1‐(biphenyl‐4‐yl)ethylidene]hydrazinylidene}methyl)‐1H‐indole (BEHI) acetonitrile disolvate, C₂₃H₁₉N₃·2CH₃CN, has been designed and synthesized. The structure of BEHI was characterized by elemental analysis, Q‐TOF (quadrupole time‐of‐flight) MS, NMR, UV–Vis and single‐crystal X‐ray diffraction. The antitumour activity of the target molecule was evaluated by the MTT method. Results indicated that BEHI exhibited rather potent cytotoxic activity against human A549 (IC₅₀ = 20.5 µM ) and mouse breast 4T₁ (IC₅₀ = 18.5 µM ) cancer cell lines. Meanwhile, to rationalize its potencies in the target, BEHI was docked into DHODH and the interactions with the active site residues were analyzed. Single‐crystal structure analysis indicated that hydrogen bonds are present only between BEHI and acetonitrile solvent molecules in the asymmetric unit. The interplay of weak π–π stacking and weak C(N)—H…π interactions between neighbouring BEHI molecules play crucial roles in the formation of the final supramolecular frameworks.     

Polydiacetylene/Anti‐HBs Complexes for Visible and Fluorescent Detection of Hepatitis B Surface Antigen on a Nitrocellulose Membrane

The immunochromatographic assay (ICA) using a nitrocellulose (NC) membrane offers several advantages. This technique is a rapid and straightforward method in contrast to other immunoassays. Polydiacetylene (PDA) vesicles have unique optical properties, displaying red color and red fluorescence at the same time. In this system, red‐phase PDA vesicles are used as a fluorescent dye as well as a surface for immobilized hepatitis B surface antibody (HBsAb). PDA has a remarkable stability compared with other fluorescent dyes. In this study, the most suitable PDA/HBsAb complexes are introduced for detecting hepatitis B surface antigen (HBsAg). Then, the PDA/HBsAb complexes affixed antibody is attached to NC membrane, which has two lines to confirm detection of HBsAg. The main advantage of this system is that the detection of HBsAg can be observed in both visible and fluorescent images due to the optical properties of polydiacetylene. Detection of HBsAg is observed up to 0.1 ng mL⁻¹ by fluorescent analysis and confirmed by red line on the NC membrane up to 1 ng mL⁻¹ (HBsAg) using the naked eye. Consequently, these results show that PDA/HBsAb complexes were successfully applied to ICA for the diagnosis of hepatitis B.     

Structural Characterization of Graphene Nanosheets for Miniaturization of Potentiometric Urea Lipid Film Based Biosensors

The present article describes a miniaturized potentiometric urea lipid film based biosensor on graphene nanosheets. Structural characterization of graphene nanosheets for miniaturization of potentiometric urea lipid film based biosensors have been studied through atomic force microscopy (AFM) and transmission electron microscopy (TEM) measurements. UV‐Vis and Fourrier transform IR (FTIR) spectroscopy have been utilized to study the pre‐ and postconjugated surfaces of graphene nanosheets. The presented potentiometric urea biosensor exhibits good reproducibility, reusability, selectivity, rapid response times (～4 s), long shelf life and high sensitivity of ca. 70 mV/decade over the urea logarithmic concentration range from 1×10^?6 M to 1×10^?3 M.     

Determination of the inhibitory effect of green tea extract on glucose‐6‐phosphate dehydrogenase based on multilayer capillary enzyme microreactor

Natural herbal medicines are an important source of enzyme inhibitors for the discovery of new drugs. A number of natural extracts such as green tea have been used in prevention and treatment of diseases due to their low‐cost, low toxicity and good performance. The present study reports an online assay of the activity and inhibition of the green tea extract of the Glucose 6‐phosphate dehydrogenase (G6PDH) enzyme using multilayer capillary electrophoresis based immobilized enzyme microreactors (CE‐IMERs). The multilayer CE‐IMERs were produced with layer‐by‐layer electrostatic assembly, which can easily enhance the enzyme loading capacity of the microreactor. The activity of the G6PDH enzyme was determined and the enzyme inhibition by the inhibitors from green tea extract was investigated using online assay of the multilayer CE‐IMERs. The Michaelis constant (K_m) of the enzyme, the IC₅₀ and K_i values of the inhibitors were achieved and found to agree with those obtained using offline assays. The results show a competitive inhibition of green tea extract on the G6PDH enzyme. The present study provides an efficient and easy‐to‐operate approach for determining G6PDH enzyme reaction and the inhibition of green tea extract, which may be beneficial in research and the development of natural herbal medicines. Copyright © 2016 John Wiley & Sons, Ltd.     

Protein-Mediated Suppression of Rolling Circle Amplification for Biosensing with an Aptamer-Containing DNA Primer

We report a method to detect proteins via suppression of rolling circle amplification (RCA) by using an appropriate aptamer as the linear primer (denoted as an aptaprimer) to initiate RCA. In the absence of a protein target, the aptaprimer is free to initiate RCA, which can produce long DNA products that are detected via binding of a fluorescent intercalating dye. Introduction of a target causes the primer region within the aptamer to become unavailable for binding to the circular template, inhibiting RCA. Using SYBR Gold or QuantiFluor dyes as fluorescent probes to bind to the RCA reaction product, it is possible to produce a generic protein-modulated RCA assay system that does not require fluorophore- or biotin-modified DNA species, substantially reducing complexity and cost of reagents. Based on this modulation of RCA, we demonstrate the ability to produce both solution and paper-based assays for rapid and quantitative detection of proteins including platelet derived growth factor and thrombin.     

Liposome-Boosted Peroxidase-Mimicking Nanozymes Breaking the pH Limit

Peroxidase-mimicking nanozymes such as Fe₃O₄ nanoparticles are promising substitutes for natural enzymes like horseradish peroxidase. However, most such nanozymes work efficiently only in acidic conditions. In this work, the influence of various liposomes on nanozyme activity was studied. By introducing negatively charged liposomes, peroxidase-mimicking nanozymes achieved oxidation of 3,3′,5,5′-tetramethylbenzidine (TMB) in neutral and even alkaline conditions, although the activity towards anionic 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) was inhibited. The Fe₃O₄ nanoparticles adsorbed on the liposomes without disrupting membrane integrity as confirmed by fluorescence quenching, dye leakage assays, and cryo-electron microscopy. Stabilization of the blue-colored oxidized products of TMB by electrostatic interactions was believed to be the reason for the enhanced activity. This work has introduced lipids to nanozyme research, and it also has practically important applications for using nanozymes at neutral pH, such as the detection of hydrogen peroxide and glucose.     

Synthesis of glycerol-thiophene nanoparticles,a suitable sensing platform for voltammetric determination of guaifenesin

Boronic acid functionalized materials have gained much attention in both chemistry and biology fields due to their multivalent covalent interactions with cis-diol containing (macro) molecules. The remarkable progress in this field has resulted in the development of their biomedical applications, such as, biosensors and nanocarriers. In this study, the spherical nanoparticles consisting of glycerol and 2,5-thiophenediylbisboronic acid were synthesized by one-pot ring opening copolymerization of a mixture of glycidol and 2,5-thiophenediylbisboronic acid. The synthesized nanoparticles were used for the modification of the glassy carbon electrode and the determination of Guaifenesin. The synthesized polymeric nanoparticles were characterized by different spectroscopic and microscopic methods including UV–vis, IR, NMR, DLS, and SEM. Additionally, the electrochemical behavior of the fabricated electrode toward Guaifenesin was investigated with cyclic voltammetry and electrochemical impedance spectroscopy.