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.     

Molecular simulation of non-equilibrium methane hydrate decomposition process

We recently performed constant energy molecular dynamics simulations of the endothermic decomposition of methane hydrate in contact with water to study phenomenologically the role of mass and heat transfer in the decomposition rate [S. Alavi, J.A. Ripmeester, J. Chem. Phys. 132 (2010) 144703]. We observed that with the progress of the decomposition front temperature gradients are established between the remaining solid hydrate and the solution phases. In this work, we provide further quantitative macroscopic and molecular level analysis of the methane hydrate decomposition process with an emphasis on elucidating microscopic details and how they affect the predicted rate of methane hydrate decomposition in natural methane hydrate reservoirs. A quantitative criterion is used to characterize the decomposition of the hydrate phase at different times. Hydrate dissociation occurs in a stepwise fashion with rows of sI cages parallel to the interface decomposing simultaneously. The correlations between decomposition times of subsequent layers of the hydrate phase are discussed.     

Accurate dispensing of volatile reagents on demand for chemical reactions in EWOD chips

Digital microfluidic chips provide a new platform for manipulating chemicals for multi-step chemical synthesis or assays at the microscale. The organic solvents and reagents needed for these applications are often volatile, sensitive to contamination, and wetting, i.e. have contact angles of <90° even on the highly hydrophobic surfaces (e.g., Teflon? or Cytop?) typically used on digital microfluidic chips. Furthermore, often the applications dictate that the processes are performed in a gas environment, not allowing the use of a filler liquid (e.g., oil). These properties pose challenges for delivering controlled volumes of liquid to the chip. An automated, simple, accurate and reliable method of delivering reagents from sealed, off-chip reservoirs is presented here. This platform overcomes the issues of evaporative losses of volatile solvents, cross-contamination, and flooding of the chip by combining a syringe pump, a simple on-chip liquid detector and a robust interface design. The impedance-based liquid detection requires only minimal added hardware to provide a feedback signal to ensure accurate volumes of volatile solvents are introduced to the chip, independent of time delays between dispensing operations. On-demand dispensing of multiple droplets of acetonitrile, a frequently used but difficult to handle solvent due to its wetting properties and volatility, was demonstrated and used to synthesize the positron emission tomography (PET) probe [(18)F]FDG reliably.     

Nonequilibrium Effects and Multiphase Flow in Porous Media

In this paper we develop a more general formulation for transient multiphase flow in porous media based on physics observed in core-scale and micromodel experiments. We account for non-equilibrium effects by considering redistribution time and treat saturation by evolving locally moving time-averages of the saturation. Several families of models arise from approximations to the general formulation with various degrees of accuracy. The classical Buckley-Leverett and Barenblatt expressions are special cases of these families. We explore the behaviors of a number of special cases arising from the proposed general formulation using established and novel numerical schemes that provide nonlinear physics-based preconditioning. The agreement observed between numerical and experimental results demonstrates the consistency of the proposed abstraction.     

A concise review on the role of nanoparticles upon the productivity of solar desalination systems

Journal of Thermal Analysis and Calorimetry - In recent years, nanofluids have been widely used to improve the performance of various energy systems due to their favourable thermo-physical and...     

Synthesis,in vitro $$\alpha $$-glucosidase inhibitory activity,and in silico study of ( E)-thiosemicarbazones and ( E)-2-(2-(arylmethylene)hydrazinyl)-4-arylthiazole derivatives

This study is focused on the identification of thiazole-based inhibitors for the \(\alpha \)-glucosidase enzyme. For that purpose, (E)-2-(2-(arylmethylene)hydrazinyl)-4-arylthiazole derivatives were synthesized in two steps and characterized by various spectroscopic techniques. All derivatives and intermediates were evaluated for their in vitro \(\alpha \)-glucosidase inhibitory activity. Thiosemicarbazones 20 and 35, and cyclized thiazole derivatives 2, 5–11, 13, 15, 21–24, 27–31, and 36–37 showed significant inhibitory potential in the range of \(\hbox {IC}_{50}=6.2\pm 0.19\)–\(43.6\pm 0.23~\upmu \hbox {M}\) as compared to standard acarbose (\(\hbox {IC}_{50}=37.7\pm 0.19~\upmu \hbox {M}\)). A molecular modeling study was carried out to understand the binding interactions of compounds with the active site of enzyme.     

Endoscopic fuel film,chemiluminescence, and soot incandescence imaging in a direct-injection spark-ignition engine

In direct-injection spark-ignition engines, fuel films formed on the piston surface due to impinging sprays are a major source of soot. Previous studies investigating the fuel films and their correlation to soot production were mostly performed in model experiments or optical engines. These experiments have different operating conditions compared to commercial engines. In this work, fuel films and soot are visualized in an all-metal engine with endoscopic access via laser-induced fluorescence (LIF) and natural incandescence, respectively. Gasoline and a mixture of isooctane/toluene were used as fuel for the experiments. The fuel films were excited by 266 nm laser pulses and visualized by an intensified CCD camera through a modular UV endoscope. Gasoline yielded much higher signal-to-noise ratio, and this fuel typically took an order of magnitude longer to evaporate than isooctane/toluene. The effects of injection time, injection pressure, engine temperature, and combustion on the fuel-film evaporation time were investigated. This film survival time was reduced with higher engine temperature, higher injection pressure, and later injection time, with engine temperature being the most significant parameter, whereas skip-fired combustion had very little effect on the film survival time. In complementary experiments, LIF from fuel films and soot incandescence were simultaneously visualized by an intensified double-frame CCD camera. At lower engine temperatures the fuel films remained distinct, and soot formation was limited to regions above the films, whereas at higher temperatures, fuel films, and hence the soot, appeared to be spread over the whole piston surface. Finally, high-speed imaging showed the spray, chemiluminescence, and soot incandescence, with results broadly consistent with fuel-film LIF and soot incandescence imaging.     

Synthesis of functionalized pyrrole derivatives via diverse cyclization of azomethine ylide and olefins

The azomethine ylides are generally used in 1,3-dipolar cycloadditions with various dipolarophiles. In this work, a new and diverse route has been developed for the azomethine ylides, for synthesis of novel pyrrole derivatives. The azomethine ylide, produced via C–H activation of unreactive C(sp³)–H bond of 2-methylquinoline, by molecular iodine, in the presence of pyridine. Herein, we represent novel pyrrole derivatives, synthesized from the reaction of pyridinium ylide with olefins, which formed via a reaction of isatin, dialkyl acetylenedicarboxylate derivatives and pyridine as a base in moderate to excellent yields. Various features of this cyclization, discussed.

Graphic abstract

     

Synthesis of tetrahydro-1,4-benzodiazepine-2-ones on hydrophilic polyamide SynPhase lanterns

Solid-phase synthesis is greatly dependent on the solid support. Here, we report the use of a new hydrophilic grafted surface on SynPhase lanterns in solid-phase organic chemistry. A convenient and facile solid-phase synthesis of disubstituted 1,4-benzodiazepine-2-ones on polyamide SynPhase lanterns is described. The key step of the synthesis involved a reduction-cyclization of a nitroaryl methyl ester with a mixture of tin(II) chloride dihydrate and ammonium acetate in water and ethanol at elevated temperature to give the desired target compounds. A library of 21 disubstituted 1,4-benzodiazepine-2-ones was prepared.