Design of a coupled bioluminescent assay for a recombinant pyruvate kinase from a thermophilic Geobacillus

A simple and rapid method using coupled bioluminescent assay was developed to determine level of ADP. ADP is involved in many biological reactions and ADP assay can be used for assaying some reactions universally by monitoring ADP formation or depletion. ADP analysis involves incubation of ADP or extracts containing ADP with pyruvate kinase (PK) and PEP. The ATP formed by this reaction is determined by measuring the intensity of the initial light flash produced when luciferin-luciferase preparation injected into the reaction mixture. In regard to the main role of the PK in this assay, the gene of PK from a Geobacillus species has been cloned in expression vector pET28a (+), sequenced and overexpressed in Escherichia coli. Recombinant protein was purified using Ni-NTA column and then the purified PK was used in a coupled bioluminescent assay for ADP measurement. Kinetic properties of PK are determined according to a bioluminescent assay using firefly luciferase.     

Reversed-phase dispersive liquid-liquid microextraction with multivariate optimization for sensitive HPLC determination of tyrosol and hydroxytyrosol in olive oil

A reversed-phase dispersive liquid-liquid microextraction (RP-DLLME) method coupled to HPLC was developed for the extraction of hydroxytyrosol (HTy) and tyrosol (Ty) from virgin olive oil. In this first application of the RP-DLLME method to non-polar samples, the phenolic compounds were directly extracted into an aqueous micro-drop, which could be injected into a chromatography column without any further pretreatment. A glass test tube with lengthened conical bottom was fitted inside a centrifuge tube in this work for more efficient withdrawal of the sedimented phase with a microsyringe. The volumes of water and ethyl acetate, the pH of water and the centrifuge time as four effective parameters on the extraction were optimized by a central composite design (response surface) method. Five replicated analyses under the optimized conditions (i.e., 0.2 mL ethyl acetate as disperser and 100 μL water at pH 11 as the extraction solvent) resulted in recoveries of 104.3 and 97.6%, and relative standard deviations of 5.75 and 4.57 for HTy and Ty, respectively. The detection limit of the method (3σ) was 0.043 mg L(-1) for HTy and 0.032 mg L(-1) for Ty. The method was successfully applied to the determination of HTy and Ty in five olive oil samples.     

Fabrication and Application of a Sensitive and Highly Stable Copper Hexacyanoferrate Modified Carbon Ionic Liquid Paste Electrode for Hydrogen Peroxide and Glucose Detection

A three‐factor mixture design and response surface methodology were employed to find the optimal weight ratio of graphite powder, n‐dodecylpyridinium hexafluorophosphate and paraffin for the fabrication of a copper hexacyanoferrate modified carbon ionic liquid paste electrode (CuHCFe‐CILPE). The fabricated sensor showed electrocatalytic activity towards oxidation and reduction of hydrogen peroxide. It also was observed that the electrocatalytic activity for hydrogen peroxide oxidation was much higher than the electrocatalytic activity for hydrogen peroxide reduction. Glucose oxidase was then successfully immobilized on the surface of the proposed sensor to examine the possibility of using CuHCFe‐CILPE for the biosensor fabrication.     

Adsorption of polyfunctional 5‐fluorouracil and 2,4‐dithio‐5‐fluorouracil on Au(111) surface: Structure,energy, and electronic transmission

Adsorption of 5‐fluorouracil (5‐FU) and 2,4‐dithio‐5‐fluorouracil (2,4‐DT‐5‐FU) on Au(111) surface at low coverage is studied by using periodic‐slab‐density functional theory calculation. Isolated 5‐FU molecule adsorbs preferentially at bridge site in a vertical configuration via N? H group by forming the N? H···Au nonconventional H‐bond. The formation of the anchor Au? O bond is not observed. Substitution of oxygen atoms of 5‐FU with sulfur strongly influences the nature of adsorption and leads to the Au? S anchor bond and the N? H···Au nonconventional H‐bond of single 2,4‐DT‐5‐FU molecule on Au(111) surface. The adsorption site and orientation of 2,4‐DT‐5‐FU molecule on the surface are similar to those of 5‐FU. The metal–molecule coupling effects at asymmetric Au/S(N? H)S/mol/C? H/Au and Au/N? H/mol/O/Au transport junctions and symmetric Au/S(N? H)S/mol/mol/S(N? H)S/Au and Au/O/mol/mol/O/Au transport junctions are also investigated. The electronic structure is analyzed in detail, and the obtained results are used for illustrating the electron transmission in metal–molecule–metal systems. © 2011 Wiley Periodicals, Inc. Int J Quantum Chem, 2011     

n-Butylammonium carboxylates/Tf2O: ionic liquid based systems for the synthesis of unsymmetrical imides via a Ritter-type reaction

We have developed a new method for the preparation of unsymmetrical imides using liquid carboxylate salts via a Ritter-type process. The reactions were carried out with nitriles and n-butylammonium carboxylates as ionic liquids in the presence of triflic anhydride (Tf₂O) as the promoter. Mild reaction conditions, simplicity of the procedure, and proton-free conditions are the main advantages of this procedure.     

On maximal commutative subrings of non-commutative rings

We observe that every non-commutative unital ring has at least three maximal commutative subrings. In particular, non-commutative rings (resp., finite non-commutative rings) in which there are exactly three (resp., four) maximal commutative subrings are characterized. If R has acc or dcc on its commutative subrings containing the center, whose intersection with the nontrivial summands is trivial, then R is Dedekind-finite. It is observed that every Artinian commutative ring R, is a finite intersection of some Artinian commutative subrings of a non-commutative ring, in each of which, R is a maximal subring. The intersection of maximal ideals of all the maximal commutative subrings in a non-commutative local ring R, is a maximal ideal in the center of R. A ring R with no nontrivial idempotents, is either a division ring or a right ue-ring (i.e., a ring with a unique proper essential right ideal) if and only if every maximal commutative subring of R is either a field or a ue-ring whose socle is the contraction of that of R. It is proved that a maximal commutative subring of a duo ue-ring with finite uniform dimension is a finite direct product of rings, all of which are fields, except possibly one, which is a local ring whose unique maximal ideal is of square zero. Analogues of Jordan-Hölder Theorem (resp., of the existence of the Loewy chain for Artinian modules) is proved for rings with acc and dcc (resp., with dcc) on commutative subrings containing the center. A semiprime ring R has only finitely many maximal commutative subrings if and only if R has a maximal commutative subring of finite index. Infinite prime rings have infinitely many maximal commutative subrings.     

A meshless like method for the approximate solution of integral equations over polygons

In this paper, a numerical scheme based on a Gauss-like cubature formula from Sammariva and Vianello (2007) [1] is introduced for approximate solution of integral equations over a polygonal domain with a piecewise straight lines boundary in R²${\mathbb{R}}^2$. The proposed technique is a meshless like method with sufficient precision, which does not require any discretization of the polygon domain or any preprocessing such as mesh refinement. The error analysis of the method is provided and some numerical experiments are also presented to evaluate the performance of the proposed algorithm.     

Microperoxidase-11/NH2-FSM16 as a H2O2-resistant heterogeneous nanobiocatalyst: a suicide-inactivation study

The catalytic activity of heme peptides is an area of intense investigation. They are utilized for exploring the fine details of structural and functional properties of an active site, and to create minimized and industrial catalysts. The peroxidase activity and kinetics of suicide-inactivation of microperoxidase-11/FSM16 as a heterogeneous nanobiocatalyst in oxidation reaction of guaiacol were studied in the presence of high concentration of hydrogen peroxide (2?mM), as its natural suicide-substrate. The substrate concentration was first-order in relation to aromatic substrate (AH), and the ratio of suicide-substrate (H₂O₂) was kept much higher than the benign substrate (guaiacol). The results of kinetic analysis confirmed a similar mechanism for suicide-peroxide inactivation of horseradish peroxidase (HRP), microperoxidase (MP-11) and MP-11/NH₂-FSM16. Inactivation kinetic parameters, including intact activity of MP-11/NH₂-FSM16, ??_i, and the apparent inactivation rate constant (k _i) were obtained as 0.229?±?0.009?min^?1 and 0.651?±?0.041?min^?1 at [H₂O₂]?=?2.0?mM, respectively, in 5.0?mM phosphate buffer solution (PBS; pH 7.0) at 27?°C. Our results indicated that covalent immobilization of microperoxidase onto NH₂-FSM16 protected the heme group against peroxide inactivation resulting in generation of an efficient peroxide-resistant heterogeneous nanobiocatalyst.     

Natural Convection Induced by a Heated Vertical Plate Embedded in a Porous Medium with Transpiration: Local Thermal Non-equilibrium Similarity Solutions

This paper is concerned with the thermal non-equilibrium free convection boundary layer, which is induced by a vertical heated plate embedded in a saturated porous medium. The effect of suction or injection on the free convection boundary layer is also studied. The plate is assumed to have a linear temperature distribution, which yields a boundary layer of constant thickness. On assuming Darcy flow, similarity solutions are obtained for governing the steady laminar boundary layer equations. The reduced Nusselt numbers for both the solid and fluid phases are calculated for a wide range of parameters, and compared with asymptotic analyses.     

Forced Convection Heat Transfer of Nanofluids in a Porous Channel

This article is concerned with the effects of flow and migration of nanoparticles on heat transfer in a straight channel occupied with a porous medium. Investigation of force convective heat transfer of nanofluids in a porous channel has not been considered completely in the literature and this challenge is generally considered to be an open research topic that may require more study. The fully developed flow and steady Darcy?CBrinkman?CForchheimer equation is employed in porous channel. The thermal equilibrium model is assumed between nanofluid and solid phases. It is assumed that the nanoparticles are distributed non-uniformly inside the channel. As a result the volume fraction distribution equation is also coupled with governing equations. The effects of parameters such as Lewis number, Schmidt number, Brownian diffusion, and thermophoresis on the heat transfer are completely studied. The results show that the local Nusselt number is decreased when the Lewis number is increased. It is observed that as the Schmidt number is increased, the wall temperature gradient is decreased and as a consequence the local Nusselt number is decreased. The effects of Lewis number, Schmidt number, and modified diffusivity ratio on the volume fraction distribution are also studied and discussed.