Enantioselective inhibition of immobilized acetylcholinesterase in biosensor determination of pesticides

Chiral effects for the inhibition of acetylcholinesterase by organophosphorus pesticides were investigated for insecticide malathion and malaoxon, which is a metabolic product of malathion in living organisms. Studies were carried out using a bienzymatic biosensor with immobilized acetylcholinesterase, choline oxidase, and with Prussian Blue used as a mediator. In both cases the R enantiomers accelerate acetylocholinesterase inhibition. The chiral effect in inhibition was much more pronounced in fast flow measurements than in batch measurements.      

Synthesis,structure and capacitive properties of cobalt hydroxide films on stainless steel substrates

Cobalt (hydro)oxide films on AISI 304 stainless steel and sintered metal fibre filter Bekipor ST 20AL3 were prepared using electrochemical deposition from neutral cobalt acetate solutions under galvanostatic conditions. Deposited films were structurally characterized using X-ray diffraction, Fourier transform infrared spectroscopy and scanning electron microscopy. All electrochemical measurements were performed in aqueous NaOH solution. Capacitive behavior of different films was evaluated using cyclic voltammetry data. The highest specific capacitance (965 F g^?1) was reached when Bekipor ST 20AL3 mesh was used as a support for electroactive substance.      

<Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> yeast immobilized on marrow stem sunflower and polyacrylamide hydrogels

Biocatalysts with microorganisms immobilized on solid carriers could provide the solution for development of continuous industrial processes for ethanol obtaining by fermentation of sugars. In this study, modified polyacrylamide hydrogels and marrow stem sunflower are used as supports for Saccharomyces cerevisiae yeast immobilization. The obtained structures are used for fermentation of molasses in batch systems. The free yeast cells are used as reference. The modification of polyacrilamide matrix with (2-hydroxyethyl)methacrylate has a positive effect on structure pore uniformity and fermentation performance. The mechanical properties of the obtained biocatalysts are compared. The novel natural matrix has net superior compression strength.

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Synthesis of terbium(III) complex with a biscoumarin derivative and its immobilization in PMMA-based composite thin films with fluorescent properties

An amorphous complex of Tb(III) with the biscoumarin derivative 3,3′-[(4-hydroxyphenyl)methylene)]bis-(4-hydroxy-2H-1-benzopyran-2-one), Tb(H₂L)₃, was successfully synthesized and characterized. IR- and ¹H-NMR-spectroscopy were used to investigate the coordination of the ligand around the Tb(III) ion. Values for the quantum yield and the life time of the excited state of the complex were obtained. The complex was immobilized in transparent and flexible PMMA-based films by a simple casting technique. PMMA/chloroform solutions were used in synthetic procedures that resulted in both glass-supported and self-supporting nanocomposite films. The morphology of the films was studied by scanning electron microscopy, atomic force microscopy and transmission electron microscopy, showing the formation of crack-free films. The presence of the Tb(III) complex in the matrix was proven by the presence of characteristic bands in the IR spectra. Fluorescence microscopy and fluorescence spectroscopy demonstrated the promising optical properties of the films showing the characteristic emission bands of the Tb(III) ions. The longer life time of the excited state of the immobilized complex confirmed the protective role of the PMMA matrix on the optical properties of the complex. The composite films possessing optical properties have the potential for application as active components in optical devices.      

Biocatalytic designs for the conversion of renewable glycerol into glycerol carbonate as a value-added product

A comparative study of two different biocatalytic models, e.g. enzyme immobilized on magnetic particles (EIMP) and cross-linking enzyme aggregates onto magnetic particles (CLEMPA) was performed. The first model was designed as enzyme-immobilized on the magnetic particles surface (EIMP). The second model was constructed as a network structure with the enzyme aggregates and magnetic particles placed into the nodes and polyglutaraldehyde cross-linker as the network ledges. The design was called cross-linking enzyme aggregates onto magnetic particles (CLEMPA). The biocatalysts were prepared using lipase enzyme from Aspergillus niger for catalyzing the glycerol (Gly) conversion to glycerol carbonate (GlyC). The biocatalyst characteristics for both designs (EIMP and CLEMPA) were evaluated using scanning electron microscopy (SEM), laser light scattering (LLS) and UV-Vis techniques. The EIMP model was strongly influenced by the composition of the polymeric layer covering the particles surface, while the size of the magnetic particles affected mostly the CLEMPA design. Also, the biocatalytic capacity of the tested models was evaluated as maximum 52% Gly conversion with 90% GlyC selectivity for EIMP, and 73% Gly conversion with 77% GlyC selectivity for CLEMPA. Both biocatalytic models were successfully used to prepare GlyC from “crude” glycerol collected directly from the biodiesel process (e.g. 49% Gly conversion with 91% GlyC selectivity for EIMP and 70% Gly conversion with 80% GlyC selectivity for CLEMPA).      

Direct electrochemical sensing of glucose using glucose oxidase immobilized on functionalized carbon nanotubes via a novel metal chelate-based affinity method

We report on a novel amperometric glassy carbon biosensing electrode for glucose. It is based on the immobilization of a highly sensitive glucose oxidase (GOx) by affinity interaction on carbon nanotubes (CNTs) functionalized with iminodiacetic acid and metal chelates. The new technique for immobilization is exploiting the affinity of Co(II) ions to the histidine and cysteine moieties on the surface of GOx. The direct electrochemistry of immobilized GOx revealed that the functionalized CNTs greatly improve the direct electron transfer between GOx and the surface of the electrode to give a pair of well-defined and almost reversible redox peaks and undergoes fast heterogeneous electron transfer with a rate constant (k _s) of 0.59?s^?1. The GOx immobilized in this way fully retained its activity for the oxidation of glucose. The resulting biosensor is capable of detecting glucose at levels as low as 0.01?mM, and has excellent operational stability (with no decrease in the activity of enzyme over a 10?days period). The method of immobilizing GOx is easy and also provides a model technique for potential use with other redox enzymes and proteins.

Chemical modification and characterization of poly(ethylene terephthalate) surfaces for collagen immobilization

The functionalization of poly(ethylene terephthalate) (PET) surface films by reactions with multifunctional amines such as triethylenetetramine (TETA), and tetraethylenepentamine (TEPA) was investigated. For the functionalization of PET films surface we used a new way of treatment, a “sandwich model”. Physical-chemical properties of functionalized PET films were analysed. Qualitative and quantitative determination of the introduced amine groups were examined by means of Fourier Transform Infrared Attenuated Total Reflexion (FTIR — ATR), X-ray photoelectron spectroscopy (XPS), and potentiometric titration. Gained wetting properties were determined by using contact angle measurements and thoroughly analysed by acid-base approach. In addition, surface topography was investigated by atomic force microscopy (AFM). The amount of the introduced amino groups after TETA incorporation has been found to be two times higher as compared to TEPA. Wetting properties were significantly improved after aminolysis. Surface free energy was higher for PET — TETA treated film than that observed for PET — TEPA treated which is in accordance with titration results. The collagen immobilization onto PET treated films was evidenced by using AFM and subsequently by using XPS.      

Comparison characterization of copper selenide thin layers prepared on polyamide 6 films by sorption-diffusion method

Some earlier synthesized copper selenide (Cu _x Se) layers formed on the surface of polyamide 6 by sorption-diffusion method using potassium selenotrithionate (K₂SeS₂O₆) as precursor of selenium were characterized by the XRD, XPS and SEM methods. According to the results of the SEM studies, the most uniform Cu _x Se layers form at the 2.5 h polyamide seleniumized duration at the temperature of 60°C. The thickness of layers, which dependeds on the duration of seleniumization, changed in the range of 0.8–3.2 µm. The XRD patterns of not previously studied Cu _x Se layers showed their phase composition of six copper selenides: Cu₂Se, two phases of CuSe₂, Cu₃Se₂, berzellianite, Cu_2-x Se, and bellidoite Cu₂Se. Analysis of the XRD and XPS data shows that the macrostructure and composition of the Cu_xSe layers depend on the conditions of formation of these layers.      

Metals content of soil,leaves and wild fruit from Serbia

The concentrations of Zn, Mn, Fe, Pb, Ni, Cu and Cd in soil, leaves and edible wild fruit (Crataegus laevigata L., Cornus mas L. and Prunus spinosa L.) from southeast Serbia were determined by atomic absorption spectroscopy. Metal translocations from soil to fruit were calculated as well as their oral intake and health risk indices. Positive correlations were found among metal concentrations in soil, leaves and fruit.      

New genetic insights to consider coffee waste as feedstock for fuel,feed, and chemicals

Caffeine is a natural plant product found in many drinks, including coffee, tea, soft and energy drinks. Due to caffeine’s presence in the environment, microorganisms have evolved two different mechanisms to live on caffeine. The genetic maps of the caffeine N-demethylation pathway and C-8 oxidation pathway have been discovered in Pseudomonas putida CBB5 and Pseudomonas sp. CBB1, respectively. These genes are the only characterized bacterial caffeine-degrading genes, and may be of great value in producing fine chemicals, biofuels, and animal feed from coffee and tea waste. Here, we present preliminary results for production of theobromine and 7-methylxanthine from caffeine and theobromine, respectively, by two strains of metabolically engineered E. coli. We also demonstrate complete decaffeination of tea extract by an immobilized mixed culture of Klebsiella and Rhodococcus cells. These processes provide a first level demonstration of biotechnological utilization of coffee and tea waste.      

Aerobic oxidation of cumene catalysed by 4-alkyloxycarbonyl-N-hydroxyphthalimide

4-Hexyloxycarbonyl-, 4-dodecyloxycarbonyl- and 4-hexadecyloxycarbonyl-N-hydroxyphthalimides were synthesised using trimellitic anhydride chloride as the starting material. The obtained lipophilic derivatives of N-hydroxyphthalimide were applied as catalysts of the cumene oxidation reaction with oxygen performed in polar acetonitrile, in non-polar tert-butylbenzene and in the absence of a solvent. The courses of reactions catalysed by N-hydroxyphthalimide and its derivatives were compared.      

Polymer conjugates with potential biological activity based on new derivatives of 2-mercaptobenzoxazole-synthesis and characterization

New potentially biologically active compounds derived from 2-mercapto-benzoxazole were synthesized and coupled on polymeric support of poly (maleic anhydride-alt-vinyl acetate) for the preparation of polymer-drug conjugates with controlled drug release. All compounds were characterized by elemental and spectroscopy (FT-IR, ¹H-NMR) analysis. The toxicological tests recommend the products for further laboratory screening.      

Salutaridine and its derivatives as thebaine-equivalents in the synthesis of aporphines

Here we report the transformation of tetracyclic morphinan salutaridine (1) into 2,3,10,11-tetrasubstituted (R)-aporphines. This method serves as another chemical proof of the previously verified biosynthetic connection with pentacyclic morphinan-6,8-diene-type thebaine. In the presence of nucleophiles, this procedure could lead to pharmacologically interesting new tetrasubstituted aporphinoids. The enantioselective synthesis of 7S-salutaridinol (2) has been also achieved in order to investigate the acid-catalyzed reactions of this natural morphinan.      

P-Triazinylphosphonium chlorides as a new group of coupling reagents

Tertiary phosphines in reaction with 2-chloro-4,6-dialkoxy-1,3,5-triazines gave unstable quaternary P-triazinylphosphonium chlorides, which readily decomposed with a departure of the alkyl group of the triazine ring substituent. Stable quaternary P-triazinylphosphonium chlorides were obtained only in reaction of 2-chloro-4,6-diphenoxy-1,3,5-triazine. Both, stable quaternary P-triazinylphosphonium chlorides as well unstable analogues prepared in situ activated carboxylic acid yielding “superactive” triazine esters, useful as highly efficient reagents in peptide bond synthesis.      

Synthesis, morphology and specific magnetization of the electrodeposited Zn-Ni-P thin films on copper substrate from non-cyanide electrolyte

Thin films of Zn-Ni-P on a copper substrate were synthesized by electrodeposition from chloride baths. It was found that the diffraction reflections of the crystal structure of Zn-Ni-P thin layers occur at thicknesses d ≥ 5 µm. The X-ray diffraction studies results confirm the formation in the Zn-Ni-P films of ZnNi₁₀P₃ compound. The morphology of the obtained films was analyzed by Scanning Electron Microscopy (SEM) and Energy Dispersive X-ray Spectrometry (EDS). The films are continuous and have a typical topography with many homogeneous globular features. EDS confirmed the formation of Zn-Ni-P coating only in 2 samples with Zn:Ni:P atomic ratios of 1:8:4 and 4:8:3, respectively. X-ray Photoelectron Spectroscopy (XPS) revealed the chemistry and the thickness of the studied thin films. At room temperature and thickness d ≥ 5 µm the investigated thin layers exhibit high values of the specific magnetizations in the range (25–37) A m² kg^?1, leading to the potential use in devices, appliances and electronics. The Curie temperature values of the synthesized Zn-Ni-P films were determined. It was found that by heating Zn-Ni-P thin layers of thicknesses d ≥ 5 µm up to a temperature T=900 K an interaction was detected with the copper substrate leading to a lower specific magnetization.      

DNA molecules site-specific immobilization and their applications

In addition to its role as a carrier of genetic information, DNA has been recognized as a construction material for the assembly of different objects and structural arrangements with nanoscale features. As a result of DNA’s self-recognition properties (based on the specific base-pairing of G-C and T-A), monolayer films of nucleic acids on solid supports have attracted an escalating attentions. Recently, numerous novel materials based on two-dimensional (2D) and three-dimensional (3D) DNA structures have been reported, which extends their utility to a large number of appliations. This review paper intends to be a new and comprehensive overview of recent strategies to site-specifically immobilized DNA on various materials, including carbonaceous substances, gold, and silica substrate, emphasizing the applications of site-specific DNA nanostructure-based devices for diagnostic, bioanalytical, food safety and environmental monitoring. Additionally, an up-to-date perspective is proposed at the end of this review.      

Amperometric determination of sulfide ion by glassy carbon electrode modified with multiwall carbon nanotubes and copper (II) phenanthroline complex

Electrocatalytic oxidation of sulfide ion on a glassy carbon electrode (GCE) modified with multiwall carbon nanotubes (MWCNTs) and a copper (II) complex was investigated. The Cu(II) complex was used due to the reversibility of the Cu(II)/Cu(III) redox couple. The MWCNTs are evaluated as a transducer, stabilizer and immobilization matrix for the construction of amperometric sensor based on Cu(II) complex adsorbed on MWCNTs immobilized on the surface of GCE. The modified GCE was applied to the selective amperometric detection of sulfide at a potential of 0.47 V (vs. Ag/AgCl) at pH 8.0. The calibration graph was linear in the concentration range of 5 µM–400 µM; while the limit of detection was 1.2 µM, the sensitivity was 34 nA µM^?1. The interference effects of SO₃ ^2?, SO₄ ^2?, S₂O₃ ^2?, S₄O₆ ^2?, Cysteine, and Cystein were negligible at the concentration ratios more than 40 times. The modified electrode is more stable with time and more easily restorable than unmodified electrode surface. Also, modified electrode permits detection of sulfide ion by its oxidation at lower anodic potentials.      

Screen-printed electrodes modified with HRP-zirconium alcoxide film for the development of a biosensor for acetaminophen detection

The development of a biosensor based on the immobilization of horseradish peroxidase (HRP) within a zirconium alkoxide-polyetilenimine film onto screen-printed electrodes (SPE) for acetaminophen detection and acetaminophen quantification in pharmaceutical products is described. The biosensor operation mode is based on monitoring the amperometric signal produced by the electrochemical reduction of the enzymatically generated electroactive oxidized species of acetaminophen in the presence of hydrogen peroxide. The enzyme immobilization is performed by retention in a polyethylenimine-zirconium alcoxide porous gel film, a technique that offers good entrapping and a protective environment for the biocomponent due to the hydration properties of the immobilization layer. SPEs have the advantage of being easily mass-produced at low costs with superior characteristics in comparison with classical electrode materials. In this configuration, zirconium alkoxide demonstrates its electrocatalytic activity. The biosensor allows the quantification of acetaminophen with a limit of detection of 6.21×10^?8 M and a linear range between 4.35×10^?7 M and 4.98×10^?6 M. Finally, the biosensor is applied to the quantitative analysis of acetaminophen in Perdolan® tablets.

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