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     

Growth and morphology of nanometer LiMn2O4 powder

The growth of Li_1+xMn₂O₄ via detonation reaction was investigated with respect to the presence of an energetic precursor, such as the metallic nitrate and the degree of confinement of the explosive charge. The detonation products were characterized by scanning electron microscopy. Powder X-ray diffraction and transmission electron microscopy were used to characterize the products. Li_1+xMn₂O₄ with 1-2 μm spherical morphology and more uniform secondary particles, but with smaller primary particles of diameters from 20 to 60 nm and a variety of morphologies were found. The oxides produced by this cheap method affirmed the validity of detonation synthesis of nano-size powders.