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.     

Preparation and properties of silicone‐containing poly(methyl methacrylate) gels

Poly(methyl methacrylate) (PMMA) gels with varying amounts of silicone and solvent and constant amounts of crosslinker were prepared by solution free radical crosslinking copolymerization of methyl methacrylate (MMA), ethylene glycol dimethacrylate (EGDM), tetraethoxysilane (TEOS) and vinyltriethoxysilane (VTES) comonomer systems. They were then studied in benzene at a total monomer concentration of 3.5 mol L^?1 and 70 °C. The conversion of monomer, volume swelling ratio, weight fraction and gel point were measured as a function of the reaction time, silicone concentration and benzene content up to the onset of macrogelation. Structural characteristics of the gels were examined by using equilibrium swelling in benzene, gel fraction and Fourier‐transform infrared (FTIR) analysis. The morphology of the copolymers was also investigated by SEM. Based on the obtained results, it was concluded that the FTIR data did not have the capacity to show the presence of the VTES or TEOS moiety in these kinds of copolymers. On the other hand, the variation of weight fraction of gel, W_g, and its equilibrium volume swelling ratio in benzene, q_v, exhibited the same behaviour as that of MMA/EGDM copolymers. Also, the dilution of the monomer mixture resulted in an increase in the gel point and swelling degree and a decrease in the percent of conversion and gel fraction. Finally, TEOS is not an ideal silicone compound for reaction in the MMA/EGDM copolymerization system, whereas VTES is a suitable silicone comonomer for this system and it has been proved useful. Copyright © 2005 Society of Chemical Industry     

Nickel and chromium cycles: Stocks and flows project part IV

Nickel and chromium are essential ingredients in alloys increasingly important for energy-efficient, environmentally friendly modern technology. Quantitative assessment of the flows of these metals through the world economy from resource extraction to final disposal informs resource policy, energy planning, environmental science, and waste management. This article summarizes the worldwide technological cycles of nickel and chromium in 2000. Stainless steel is the major use of these metals, but they serve numerous other special needs, as in superalloys for high-temperature service, as plating materials, and in coinage. Because they are used primarily in alloys, novel recycling issues arise as their use becomes more widespread. “... the great New York and St. Louis double track, nickel plated railroad...” — Norwalk, Ohio, Chronicle 10 March 1881 announces arrival of surveyors for the future Nickel Plate Railway “Later [1911] I formed an alloy of Iron and Chromium, which showed remarkable resistance to rust and tarnish ... [It was] rediscovered by an Englishman named Brearley, in 1914.” —Elwood Haynes to Stephen F. Roberts, 17 January 1925     

Influence of the silicon surface treatment by plasma etching and scratching on the nucleation of diamond grown in HFCVD - a comparative study

A comparative study for the nucleation of diamond was carried out using surface treatment like (i) surface scratching with 1 μm diamond paste and (ii) surface etching using chlorine plasma at different RF powers (50, 100 and 150 W). Atomic force microscopic study shows variation in roughness from 31 nm to 110 nm. Scratching results in random scratches, whereas plasma etches a surface uniformly. Scanning electron microscopic observations show well faceted crystallites with a predominance of angular shaped grains corresponding to 〈100〉 and 〈110〉 crystallite surfaces for the scratched as well as plasma etched substrate. Surface etching at 150 W plasma power results in a better growth in comparison with 50 and 100 W plasma powers. Chlorine-radical is found responsible for the changes in the growth morphology. Raman spectroscopy shows a sharp peak at 1,332 cm⁻¹ and a peak at ∼1,580 cm⁻¹ for both samples.     

Geotechnical properties of the soft soil in Guangzhou College City

The paper describes some geotechnical properties of the soft soils in Guangzhou College City, China, which are difficult to both sample and test. Laboratory and in situ tests were carried out to assess the physical and mechanical indices of the soils. The data were statistically analyzed and linear regression undertaken such that equations could be developed by which the geotechnical properties of the soft soils can be predicted. The statistical validity of the degree of correlation confirmed that using these equations, the mechanical indices can be estimated from physical indices determined by routine testing.