Rapid detection of organophosphorous (OP) compounds such as paraoxon would allow taking immediate decision on efficient decontamination procedures and could prevent further damage and potential casualties. In the present study, a biosensor based on nanomagnet-silica core-shell conjugated to organophosphorous hydrolase (OPH) enzyme was designed for detection of paraoxon. Coumarin1, a competitive inhibitor of the OPH enzyme, was used as a fluorescence-generating molecule. Upon excitation of cumarin1 located at the active site of the enzyme, i.e., OPH, the emitted radiations were intensified due to the mirroring effect of the nanomagnet-silica core-shell conjugated to the enzyme. In presence of paraoxon and consequent competition with the fluorophore in occupying enzyme's active site, a significant reduction in emitted radiations was observed. This reduction was proportional to paraoxon concentration in the sample. The method worked in the 10- to 250-nM concentration range had a low standard deviation (with a coefficient of variation (CV) of 6-10%), and the detection limit was as low as 5 × 10(-6) μM.

Organophosphorus (OP) compounds are one of the most hazardous chemicals used as insecticides/pesticide in agricultural practices. A large variety of OP compounds are hydrolyzed by organophosphorus hydrolases (OPH; EC 3.1.8.1). Therefore, OPHs are among the most suitable candidates that could be used in designing enzyme-based sensors for detecting OP compounds. In this work, a novel nanobiosensor for the detection of paraoxon was designed and fabricated. More specifically, OPH was covalently embedded onto chitosan and the enzyme-chitosan bioconjugate was then immobilized on negatively charged gold nanoparticles (AuNPs) electrostatically. The enzyme was immobilized on AuNPs without chitosan as well, to compare the two systems in terms of detection limit and enzyme stability under different pH and temperature conditions. Coumarin 1, a competitive inhibitor of the enzyme, was used as a fluorogenic probe. The emission of coumarin 1 was effectively quenched by the immobilized Au-NPs when bound to the developed nanobioconjugates. However, in the presence of paraoxon, coumarin 1 left the nanobioconjugate, leading to enhanced fluorescence intensity. Moreover, compared to the immobilized enzyme without chitosan, the chitosan-immobilized enzyme was found to possess decreased Km value by more than 50%, and increased Vmax and Kcat values by around 15% and 74%, respectively. Higher stability within a wider range of pH (2-12) and temperature (25-90°C) was also achieved. The method worked in the 0 to 1050 nM concentration ranges, and had a detection limit as low as 5 × 10(-11) M.

Structural and morphological changes as well as corrosion behavior of N + implanted Al in 0.6 M NaCl solution as function of N + fluence are investigated. The x-ray diffraction results confirmed AlN formation. The atomic force microscope (AFM) images showed larger grains on the surface of Al with increasing N + fluence. This can be due to the increased number of impacts of N + with Al atoms and energy conversion to heat, which increases the diffusion rate of the incident ions in the target. Hence, the number of the grain boundaries is reduced, resulting in corrosion resistance enhancement. Electrochemical impedance spectroscopy (EIS) and polarization results showed the increase of corrosion resistance of Al with increasing N + fluence. EIS data was used to simulate equivalent electric circuits (EC) for the samples. Strong dependence of the surface morphology on the EC elements was observed. The scanning electron microscope (SEM) analysis of the samples after corrosion test also showed that the surfaces of the implanted Al samples remain more intact relative to the untreated Al sample, consistent with the EIS and polarization results.

No abstract

Organophosphorus (OP) compounds are extensively used in agricultural practice for pest management. However, their residues have a long half-life in the ecosystem as well as in the agro-products, posing a serious threat to human and animal health. Aryldialkylphosphatase (EC 3.1.8.1) is widely used in detoxification procedures. In the present study, aryldialkylphosphatase was immobilised on synthesised cross-linked nano-sized gel particles, also known as nanogels, in order to enhance the enzyme's physicochemical properties. Accordingly, a new nanogel consisting of chitosan and myristic acid (CMA nanogel) was synthesised and characterised by way of Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The aryldialkylphosphatase-CMA nanogel conjugate was then assayed by FTIR, and its physicochemical characteristics were also investigated. The data obtained from SEM and TEM showed the nanogels to be homogenous spherical particles less than 50 nm in diameter. The proper formation of the nanogel and nanobioconjugate was also confirmed by FTIR spectra. In comparison with the free enzyme, the pH and thermal stability of the aryldialkylphosphatase were enhanced by the covalent immobilisation. Moreover, the immobilised enzyme could maintain approximately half of its activity over more than one month. The kinetic parameters of the aryldialkylphosphatase-CMA nanogel conjugate were also shown to undergo remarkable improvements, hence the synthesised CMA-nanogel could act as a promising support for aryldialkylphosphatase immobilisation. It is suggested that the aryldialkylphosphatase-CMA nanogel could be used for detoxifying paraoxon; a nerve agent. Further clinical experiments are underway.