Purification and renaturation of recombinant human lymphotoxin (tumour necrosis factor beta) expressed in Escherichia coli as inclusion bodies

High level expression of recombinant human tumour necrosis factor β (rh TNF-β) in Escherichia coli results in the formation of two portions of protein, namely soluble active protein and insoluble protein which is inactive and aggregates in the form of inclusion bodies (IBs). In this study, a procedure for purification and renaturation of rh TNF-β from inclusion bodies has been designed and verified experimentally with a product purity of more than 90% and a recovery of about 30%. The procedure includes washing of IBs with specific wash buffer (Triton X-100/EDTA/lysozyme/PMSF), their solubilization with 8 mol dm^?3 alkaline urea, purification with ion-exchange columns, refolding with renaturation buffer and finally concentration and desalination with an ultrafiltration membrane. The characteristics of the renatured protein were identical with those of purified protein from the soluble fraction as demonstrated by (1) SDS-PAGE, (2) cytotoxic activity on mouse L929 cells, (3) N-terminal amino acid sequence, and (4) gel filtration chromatography.     

TEM method for rapid and precise evaluation of continuous recrystallization

With the aid of a double-tilt holder in a transmission electron microscope, a new method for a rapid and precise determination of the misorientations of a large number of subgrain or grain boundaries is given in this article. By use of the method, the continuous recrystallization can be rapidly and precisely evaluated when compared to the other conventional methods.     

A solenoid-derived wiggler

A novel wiggler design for use in free-electron lasers (FELs) is proposed, consisting of a staggered array of magnetic poles situated inside the bore of a solenoid. The resultant field pattern consists of a periodic transverse magnetic field on axis, as well as a longitudinal guide field. Such a wiggler has several advantages: the longitudinal field acts to confine the electrons near the FEL axis, high fields can be attained at short wiggler periods, the field strength is easily varied, and fabrication and testing of the wiggler are relatively easy. It is planned to use this wiggler design in a far infrared FEL to be built at Stanford University     

Variational principle of combined energy—Mechanics of laminated structures

The conventional approaches for stress analysis of composite laminates have encountered discontinuity problems. In this paper, the C¹ continuity of displacements in the in-plane directions and C⁰ continuity of displacement along the thickness direction are discussed. Also, the global continuity of transverse stresses and local continuity of in-plane stresses are examined. Then, the formulations of stress analysis in both differential equation form and variational functional form are presented.     

Two-level optimal output feedback stabilizer design

Design of two-level system stabilizers is considered using an optimal reduced-order model whose state variables are torque angles and speeds. The reduced-order model retains their physical meaning and is used to design a two-level linear feedback controller that takes into account the realities and constraints of the electrical power systems. The two-level control strategy is used, and a global control signal is generated from the output variables to minimize the effect of interaction. The effectiveness of this controller is evaluated, and an example, the multimachine system, is given to illustrate the advantages of the proposed method. The responses of the system with the two-level scheme and optimal reduced order scheme are included for comparative analyses     

Effect of polymerization conditions on o‐phenylenediamine and o‐phenetidine oxidative copolymers

A series of new o‐phenylenediamine (OPD)/o‐phenetidine (PHT) copolymers with partly phenazine‐like structures has been successfully synthesized at three polymerization temperatures by chemically oxidative polymerization in four different polymerization media. The molecular structures and properties of the resulting OPD/PHT polymers were investigated by IR, UV–vis and high‐resolution ¹H NMR spectroscopies, and DSC, in order to ascertain the effect of reaction temperature, comonomer ratio and acid medium. The copolymerization mechanism of OPD with PHT monomers has been proposed. It is found that the statistical OPD/PHT copolymer obtained at a temperature of 118 °C has a higher degree of polymerization than that obtained at 12–17 °C. The OPD content in the copolymers calculated from NMR spectroscopic analysis is higher than that in the feed OPD content, whereas the OPD content calculated from element analysis is slightly lower than the feed OPD content. It can be predicted that denitrogenation takes place in the OPD units during the polymerization process at OPD/PHT molar ratios of 90/10 and 100/0. These OPD/PHT copolymers exhibit a much better solubility than the OPD homopolymer, hence suggesting an incorporation of PHT units into the phenazine structure of the homopolymer. The thermal behavior of the copolymers was also studied. Copyright © 2004 Society of Chemical Industry     

Phase composition and properties of solid solutions of GdFeO3–GdInO3 bulks

Solid solutions of the GdFeO₃–GdInO₃ system were prepared at 1550 °C by ceramic powder processing. The formulated composition was Gd(Fe_1−xIn_x)O₃ (GFI) with the indium contents at x = 0, 0.25, 0.5, 0.75, and 1.0. A stable phase of Gd(Fe_1/3In_2/3)O₃ in our system was identified by X-ray diffraction and phase composition analysis. Multi-phase morphologies were observed for GFI bulks with x = 0.5 and 0.75. Dielectric and electrical properties of the GFI bulks were investigated. The addition of 25% In³⁺ in GdFeO₃ had an obvious enhancement in polarization and led to an elevated resonance frequency. Dielectric properties of GFI bulks except GdInO₃ were strongly dependent upon the test frequency, which corresponded to the response of polarization mechanism. GdInO₃ displayed as a stable dielectric, which was frequency- and temperature-insensitive. GdInO₃ was thermally activated and became leaky until above 600 °C.     

Photocatalytic degradation of 2,2‐bis(4‐hydroxy‐3‐methylphenyl) propane (BPP) based on molecular recognition interaction

BACKGROUND: Endocrine disruptors in the aquatic environment and their potential adverse effects are currently issues of concern. One of these endocrine disruptors is 2,2‐bis(4‐hydroxy‐3‐methylphenyl)propane (BPP). In this work the molecular recognition interaction of BPP with β‐cyclodextrin (β‐CD) was studied using IR spectroscopy and steady state fluorescence spectroscopy, and the photocatalytic degradation behaviour of BPP based on molecular recognition interaction was investigated in a TiO₂/UV–visible (λ_max = 365 nm) system. This might provide a new method for the treatment of some organic pollutants in wastewater. RESULTS: β‐CD reacts with BPP to form a 1:1 inclusion complex, the formation constant of which is 4.94 × 10³ L mol^?1. The photodegradation rate constant of BPP after molecular recognition by β‐CD showed a 1.42‐fold increase in the TiO₂/UV–visible (λ_max = 365 nm) system. The photodegradation of BPP depended on the concentration of β‐CD, the pH value, the gaseous medium and the initial concentration of BPP. The photodegradation efficiency of BPP with molecular recognition was higher than that without molecular recognition. After 100 min of irradiation the mineralisation efficiency of BPP after molecular recognition by β‐CD reached 94.8%, whereas the mineralisation efficiency of BPP before molecular recognition by β‐CD was only 40.6%. CONCLUSION: The photocatalytic degradation of BPP after molecular recognition by β‐CD can be enhanced in the TiO₂/UV‐visible (λ_max = 365 nm) system. This enhancement is dependent on the enhancement of the adsorption of BPP, the moderate inclusion depth of BPP in the β‐CD cavity and the increase in the frontier electron density of BPP after molecular recognition. Copyright © 2008 Society of Chemical Industry     

Recovery of pyruvic acid with weakly basic polymeric sorbents

BACKGROUND: Carboxylic acids are among the most important substances that can be manufactured from biomass. However, the recovery of carboxylic acids from fermentation broths presents a challenging separation problem. To avoid the production of waste salts and net consumption of chemicals in the calcium carboxylate salt process, the use of reversible chemical complexation with polymeric sorbents and extractants is attractive for carboxylic acid recovery. Pyruvic acid is widely used in the manufacture of medicines, pesticides and foodstuffs and can be produced by fermentation. Since the acidity of pyruvic acid (pK_a = 2.49) is stronger than that of normal carboxylic acids, and as few reports on the recovery of pyruvic acid are available, the sorption of pyruvic acid from aqueous solution on two types of weakly basic polymeric sorbent, tertiary amine D301R and primary amine D392, was investigated over a wide pH range and at various salt (MgSO₄) concentrations. RESULTS: Overloading adsorption of pyruvic acid on both weakly basic polymeric sorbents occurred, with the overloading of D392 being greater than that of D301R. The adsorption of pyruvic acid on both sorbents was greatly affected by the solution pH and the salt concentration in the aqueous phase. An overloading model was able to predict the experimental uptake data very well. CONCLUSION: Solution pH is one of the most important operating conditions, and both polymeric sorbents D392 and D301R can be used to recover pyruvic acid from dilute aqueous solution with high efficiency at a solution pH around 2. The uptake by D392 is greater than that by D301R owing to steric hindrance of the tertiary amine. Copyright © 2008 Society of Chemical Industry