Purification and Partial Characterization of Microsomal NADH-Cytochrome b5 Reductase from Higher Plant Catharanthus roseus

A simple three step procedure was used to purify microsomal NADH-cytochrome b5 (ferricyanide) reductase to homogeneity from the higher plant C. roseus. The microsomal bound reductase was solubilized using zwitterionic detergent-CHAPS. The solubilized reductase was subjected to affinity chromatography on octylamino Sepharose 4B, blue 2-Sepharose CL-6B and NAD+-Agarose. The homogeneous enzyme has an apparent molecular weight of 33,000 as estimated by SDS-PAGE. The purified enzyme catalyzes the reduction of purified cytochrome b5 from C. roseus in the presence of NADH. The reductase also readily transfers electrons from NADH to ferricyanide (Km 56 μM), 2,6-dichlorophenolindophenol (Km 65 μM) and cytochrome Image via cytochrome b5 but not to menadione.

Theoretical and experimental investigation of framed structure-layered soil interaction problems

The plane stress solution for the interaction analysis of a framed structure, with a foundation beam, resting on a layered soil has been studied using both theoretical and photoelastic methods. The theoretical analysis has been done by using a combined analytical and finite element method. In this, the analytical solution has been used for the semi-infinite layered medium and finite element method for the framed structure. The experimental investigation has been carried out using two-dimensional photoelasticity in which modelling of the layered semi-infinite plane and a method to obtain contact pressure distribution have been discussed. The theoretical and experimental results in respect of contact pressure distribution between the foundation beam and layered soil medium, the fibre stresses in the foundation beam and framed structure have been compared. These results have also been compared with theoretical results obtained by idealizing the layered semi-infinite plane as (a) a Winkler model and (b) an equivalent homogeneous semi-infinite medium

Large negative magnetoresistance and metal-insulator transition observed in MnO/C composite coatings grown on ceramic alumina by metalorganic chemical vapor deposition

MnO/C composite coatings were grown by the metalorganic chemical vapor deposition process on ceramic alumina in argon ambient. Characterization by various techniques confirms that these coatings are homogeneous composites comprising nanometer-sized MnO particles embedded in a matrix of nanometer-sized graphite. Components of the MnO/C composite coating crystalline disordered, but are electrically quite conductive. Resistance vs. temperature measurements show that coating resistance increases exponentially from a few hundred ohms at room temperature to a few megaohms at 30 K. Logarithmic plots of reduced activation energy vs. temperature show that the coating material undergoes a metal-insulator transition. The reduced activation energy exponent for the film under zero magnetic field was 2.1, which is unusually high, implying that conduction is suppressed at much faster rate than the Mott or the Efros-Shklovskii hopping mechanism. Magnetoconductance us. magnetic field plots obtained at various temperatures show a high magnetoconductance (similar to 28.8%) at 100 K, which is unusually large for a disordered system, wherein magnetoresistance is attributed typically to weak localization. A plausible explanation for the unusual behavior observed in the carbonaceous disordered composite material is proposed. (C) 2010 Elsevier Ltd. All rights reserved.

Coupled electrostatic-elastic analysis for topology optimization using material interpolation

In this paper, we present a novel analytical formulation for the coupled partial differential equations governing electrostatically actuated constrained elastic structures of inhomogeneous material composition. We also present a computationally efficient numerical framework for solving the coupled equations over a reference domain with a fixed finite-element mesh. This serves two purposes: (i) a series of problems with varying geometries and piece-wise homogeneous and/or inhomogeneous material distribution can be solved with a single pre-processing step, (ii) topology optimization methods can be easily implemented by interpolating the material at each point in the reference domain from a void to a dielectric or a conductor. This is attained by considering the steady-state electrical current conduction equation with a `leaky capacitor' model instead of the usual electrostatic equation. This formulation is amenable for both static and transient problems in the elastic domain coupled with the quasi-electrostatic electric field. The procedure is numerically implemented on the COMSOL Multiphysics (R) platform using the weak variational form of the governing equations. Examples have been presented to show the accuracy and versatility of the scheme. The accuracy of the scheme is validated for the special case of piece-wise homogeneous material in the limit of the leaky-capacitor model approaching the ideal case.

Texture Evolution in Boron Modified Ti-6Al-4V Alloy

Owing to their high strength-to-weight ratio, excellent mechanical properties and corrosion resistance, titanium (Ti) and its alloys, especially (alpha+beta) alloys like Ti-6Al-4V is the backbone materials for aerospace, energy, and chemical industries. Trace boron addition (similar to 0.1 wt. %) to the alloy Ti-6Al-4V produces a reduction in as-cast grain size by roughly an order of magnitude resulting in enhanced ductility, higher stiffness, strength and good fracture resistance. Boron addition could also affect the evolution of texture and microstructure in the material. The solidification microstructures of Boron free as well as Boron containing Ti-6Al-4V are found to be almost homogeneous from periphery towards the center of as-cast ingot in terms of both alpha-colony size and distribution. Boron addition substantially reduces alpha-colony size (similar to 50-80 mu m). A gradual change in alpha texture from periphery towards the center has been observed with orientations close to specific texture components suggesting the formation of texture zones. The mechanism of texture evolution can be visualized as a result of variant selection during solidification through (alpha+beta) phase field.

Rapid purification of tRNA(Lys) from rat liver

Fast protein liquid chromatography (FPLC) system using Mono Q (HR 5/5) anion-exchange column chromatography followed by highly cross-linked urea-polyacrylamide gel electrophoresis (urea-PAGE) was used for the purification of lysine-specific tRNA (tRNA(Lys)) from rat liver. Crude tRNA from rat liver was fractionated with a linear gradient of NaCl (0.3-0.8 M) in triethanolamine-HCl buffer, pH 4.5, and the activity of tRNA(Lys) was found to elute between 0.51 and 0.57 M NaCl. Using this concentration range of NaCl, tRNA(Lys) was refractionated on the same column with a shallow gradient, where a single peak of tRNA(Lys) activity was obtained. tRNA(Lys)-rich fractions recovered from the second run were electrophoretically separated on 16% polyacrylamide-7 M urea gel into one major band and three minor bands. The major band showed a specific activity of 997 pmols/A260 U for tRNALys with a 43-fold purification and approximately 17% recovery. The minor bands displayed negligible or no activity for lysine. tRNA(Lys) obtained by this method was found to be homogeneous by competitive aminoacylation. The advantages of FPLC followed by urea-PAGE in the purification of an amino acid-specific tRNA over conventional column chromatography are discussed.

Sonochemical reaction engineering

Ultrasound has been widely used by chemists to enhance yields as well as rates of homogeneous as well as heterogeneous chemical reactions. The effect of ultrasound on the course of chemical reactions is mediated through cavitation bubbles it generates. High temperatures and pressures are attained inside the cavitating bubbles when they collapse. The extreme conditions so generated lead to the formation of reactive intermediates, e.g., free radiacls, inside the bubbles, which cause chemical reactions to occur when they enter the surrounding liquid. This is the mechanism through which ultrasound influences the path of homogeneous reactions. The cavitation bubbles collapse asymmetrically in the vicinity of solids, e.g., catalyst particles. Asymmetric collapse lead to formation of high speed microjets. The microjets can enhance transport rates, the increase surface area through pitting as well as particle fragmentation through collisions. Both can alter the rates of heterogeneous reaction rates. It however appears that these effects do not exhaust the scope of the influence of ultrasound on heterogeneous reactions. Modelling and quantitative prediction of the effect of ultrasound on chemical reactions is however at a stage of infancy as the phenomena are complex. Only a few examples of modelling exist in literature. Apart from this, reactor design and scaleup pose significant problems. Thus sonochemical reaction engineering offers large scope for research and development efforts.

Social selection and the evolution of cooperative groups: The example of the cellular slime moulds

In social selection the phenotype of an individual depends on its own genotype as well as on the phenotypes, and so genotypes, of other individuals. This makes it impossible to associate an invariant phenotype with a genotype: the social context is crucial. Descriptions of metazoan development, which often is viewed as the acme of cooperative social behaviour, ignore or downplay this fact. The implicit justification for doing so is based on a group-selectionist point of view. Namely, embryos are clones, therefore all cells have the same evolutionary interest, and the visible differences between cells result from a common strategy. The reasoning is flawed, because phenotypic heterogeneity within groups can result from contingent choices made by cells from a flexible repertoire as in multicellular development. What makes that possible is phenotypic plasticity, namely the ability of a genotype to exhibit different phenotypes. However, co-operative social behaviour with division of labour requires that different phenotypes interact appropriately, not that they belong to the same genotype, or have overlapping genetic interests. We sketch a possible route to the evolution of social groups that involves many steps: (a) individuals that happen to be in spatial proximity benefit simply by virtue of their number; (b) traits that are already present act as preadaptations and improve the efficiency of the group; and (c) new adaptations evolve under selection in the social context-that is, via interactions between individuals-and further strengthen group behaviour. The Dictyostelid or cellular slime mould amoebae (CSMs) become multicellular in an unusual way, by the aggregation of free-living cells. In nature the resulting group can be genetically homogeneous (clonal) or heterogeneous (polyclonal); in either case its development, which displays strong cooperation between cells (to the extent of so-called altruism) is not affected. This makes the CSMs exemplars for the study of social behaviour.

Hydrography and circulation in the western Bay of Bengal during the northeast monsoon

The Bay of Bengal, a semienclosed tropical basin that comes under the influence of monsoonal wind and freshwater influx, is distinguished by a strongly stratified surface layer and a seasonally reversing circulation. We discuss characteristics of these features in the western Bay during the northeast monsoon, when the East India Coastal Current (EICC) flows southward, using hydrographic data collected during December 1991. Vertical profiles show uniform temperature and salinity in a homogeneous surface layer, on average, 25 m deep but shallower northward and coastward. The halocline, immediately below, is approximately 50 m thick; salinity changes by approximately 3 parts per thousand. About two thirds of the profiles show temperature inversions in this layer. Salinity below the halocline hardly changes, and stratification is predominantly due to temperature variation, The halocline is noticeably better developed and the surface homogeneous layer is thinner in a low-salinity plume that hugs the coastline along the entire east coast of India, The plume is, on average, 50 km wide, with isohalines sloping down toward the coast. Most prominent in the geostrophic velocity field is the equatorward EICC. Its transport north of about 13 degrees N, computed with 1000 dbar as the level of reference, varies between 2.6 and 7.1 x 10(6) m(3) s(-1); just south of this latitude, a northwestward flow from offshore recurves and merges with the coastal current. At the southern end of the region surveyed, the transport is 7.7 x 10(6) m(3) s(-1). Recent model studies lead us to conclude that the EICC during the northeast monsoon is driven by winds along the east coast of India and Ekman pumping in the interior bay. In the south, Ekman pumping over the southwestern bay is responsible for the northwestward flow that merges with the EICC.

Identification of the active-site peptide of 2,3-dihydroxybenzoic acid decarboxylase from Aspergillus oryzae

The non-oxidative decarboxylation of aromatic acids is a poorly understood reaction. The transformation of 2,3-dihydroxybenzoic acid to catechol in the fungal metabolism of indole is a prototype of such a reaction. 2,3-Dihydroxybenzoic acid decarboxylase (EC 4.1.1.46) which catalyzes this reaction was purified to homogeneity from anthranilate induced cultures of Aspergillus oryzae using affinity chromatography. The enzyme did not require cofactors like NAD(+), PLP, TPP or metal ions for its activity. There was no spectral evidence for the presence of enzyme bound cofactors. The preparation, which was adjudged homogeneous by the criteria of SDS-PAGE, sedimentation analysis and N-terminal analysis, was characterized for its physicochemical and kinetic parameters. The enzyme was inactivated by group-specific modifiers like diethyl pyrocarbonate (DEPC) and N-ethylmaleimide (NEM). The kinetics of inactivation by DEPC suggested the presence of a single class of essential histidine residues, the second order rate constant of inactivation for which was 12.5 M(-1) min(-1). A single class of cysteine residues was modified by NEM with a second order rate constant of 33 M(-1) min(-1). Substrate analogues protected the enzyme against inactivation by both DEPC and NEM, suggesting the Location of the essential histidine and cysteine to be at the active site of the enzyme. The incorporation of radiolabelled NEM in a differential labelling experiment was 0.73 mol per mol subunit confirming the presence of a single essential cysteine per active-site. Differentially labelled enzyme was enzymatically cleaved and the peptide bearing the label was purified and sequenced. The active-site peptide LLGLAETCK and the N-terminal sequence MLGKIALEEAFALPRFEEKT did not bear any similarity to sequences reported in the Swiss-Prot Protein Sequence Databank, a reflection probably of the unique primary structure of this novel enzyme. The sequences reported in this study will appear in the Swiss-Prot Protein Sequence Databank under the accession number P80402.

Microwave-assisted selective deoxygenation of layer- and chain-containing oxides

Very rapid (within 5 min), selective, single-step deoxygenation of layer- and chain-containing oxides, MoO3, CrO3, V2O5, alpha-VOPO4 . 2H(2)O and Ag6Mo10O33 has been accomplished using graphitic carbon in a microwave-assisted reaction. The products were found to be MoO2, Cr2O3, VO2, VPO4 and a mixture of (Ag + MoO2), respectively. Products were characterised by X-ray diffraction (XRD), differential scanning calorimetry (DSC), IR and electron paramagnetic resonance (EPR) spectroscopies. Although conventional methods of preparing these materials are tedious, the present method is simple, fast and yields very homogeneous products of good crystallinity. Our results reveal that while layer- and chain-containing oxides undergo rapid microwave-assisted carbothermal reduction, the non-layered materials do not. The high structural selectivity of these reactions is suggestive of the topochemical nature of the fast reduction process.

Variability of sif and cod of stochastic structural systems

Stochastic structural systems having a stochastic distribution of material properties and stochastic external loadings in space are analysed when a crack of deterministic size is present. The material properties and external loadings are considered to constitute independent, two-dimensional, univariate, real, homogeneous stochastic fields. The stochastic fields are characterized by their means, variances, autocorrelation functions or the equivalent power spectral density functions, and scale fluctuations. The Young's modulus and Poisson's ratio are treated to be stochastic quantities. The external loading is treated to be a stochastic field in space. The energy release rate is derived using the method of virtual crack extension. The deterministic relationship is derived to represent the sensitivities of energy release rate with respect to both virtual crack extension and real system parameter fluctuations. Taylor series expansion is used and truncation is made to the first order. This leads to the determination of second-order properties of the output quantities to the first order. Using the linear perturbations about the mean values of the output quantities, the statistical information about the energy release rates, SIF and crack opening displacements are obtained. Both plane stress and plane strain cases are considered. The general expressions for the SIF in all the three fracture modes are derived and a more detailed analysis is conducted for a mode I situation. A numerical example is given.

Lamination-dependent shear deformation models for bending of angle-ply laminated plates

Lamination-dependent shear corrective terms in the analysis of bending of laminated plates are derived from a priori assumed linear thicknesswise distributions for gradients of transverse shear stresses by using CLPT inplane stresses in the two in-plane equilibrium equations of elasticity in each ply. In the development of a general model for angle-ply laminated plates, special cases like cylindrical bending of laminates in either direction, symmetric laminates, cross-ply laminates, antisymmetric angle-ply laminates, homogeneous plates are taken into consideration. Adding these corrective terms to the assumed displacements in (i) Classical Laminate Plate Theory (CLPT) and (ii) Classical Laminate Shear Deformation Theory (CLSDT), two new refined lamination-dependent shear deformation models are developed. Closed form solutions from these models are obtained for antisymmetric angle-ply laminates under sinusoidal load for a type of simply supported boundary conditions. Results obtained from the present models and also from Ren's model (1987) are compared with each other.

Novel materials synthesis by mechanical alloying/milling

An account is given of the research that has been carried out on mechanical alloying/milling (MA/MM) during the past 25 years. Mechanical alloying, a high energy ball milling process, has established itself as a viable solid state processing route for the synthesis of a variety of equilibrium and non-equilibrium phases and phase mixtures. The process was initially invented for the production of oxide dispersion strengthened (ODS) Ni-base superalloys and later extended to other ODS alloys. The success of MA in producing ODS alloys with better high temperature capabilities in comparison with other processing routes is highlighted. Mechanical alloying has also been successfully used for extending terminal solid solubilities in many commercially important metallic systems. Many high melting intermetallics that are difficult to prepare by conventional processing techniques could be easily synthesised with homogeneous structure and composition by MA. It has also, over the years, proved itself to be superior to rapid solidification processing as a non-equilibrium processing tool. The considerable literature on the synthesis of amorphous, quasicrystalline, and nanocrystalline materials by MA is critically reviewed. The possibility of achieving solid solubility in liquid immiscible systems has made MA a unique process. Reactive milling has opened new avenues for the solid state metallothermic reduction and for the synthesis of nanocrystalline intermetallics and intermetallic matrix composites. Despite numerous efforts, understanding of the process of MA, being far from equilibrium, is far from complete, leaving large scope for further research in this exciting field.

Violin string shape functions for finite element analysis of rotating Timoshenko beams

Violin strings are relatively short and stiff and are well modeled by Timoshenko beam theory. We use the static part of the homogeneous differential equation of violin strings to obtain new shape functions for the finite element analysis of rotating Timoshenko beams. For deriving the shape functions, the rotating beam is considered as a sequence of violin strings. The violin string shape functions depend on rotation speed and element position along the beam length and account for centrifugal stiffening effects as well as rotary inertia and shear deformation on dynamic characteristics of rotating Timoshenko beams. Numerical results show that the violin string basis functions perform much better than the conventional polynomials at high rotation speeds and are thus useful for turbo machine applications. (C) 2011 Elsevier B.V. All rights reserved.