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.     

Ion-Pairing of Dyclonine with Dyes

This potential for ion-pairing between dyclonine, a local anesthetic, and pharmaceutical dyes commonly used in liquid pharmaceutical formulations was examined. Occurrence of ion-pairing at a pH of 3 was confirmed by shift of the absorption maximum of all five sulfonate-containing dyes, and by measurements of octanol/water partition coefficient and surface tension. There was also an increase in the uptake of dyclonine by phosphatidylcholine liposomes in the presence of a dye (tartrazine) at pH 3 but not at pH 7, where the uncharged anesthetic is dominant.     

Infection and antibiotic therapy in 4000 burned patients treated in Milan, Italy, between 1976 and 1988

The pathogenic flora, isolated from burn wounds of patients admitted to a burn care unit during the years between 1976 and 1988 were typed and the in vitro susceptibility to antibacterial agents was recorded. Between 1976 and 1988 the general therapeutic approach was changed three times, in congruence with the prevalent nosocomial bacterial resistance. The most frequent isolates were: Pseud. aeruginosa, Staphylococcus aureus, Enterococcus spp., Proteus mirabilis, Escherichia coli, Enterobacter cloacae, Klebsiella spp. and other Enterobacteriaceae, such as Acinetobacter, Citrobacter. The most striking finding was the increase in antibiotic-resistant Enterococcus isolates. Staph. aureus, Klebsiella and E. cloacae showed susceptibility to cephalosporins, imipenem, pefloxacin, vancomycin; Enterococcus susceptibility to pefloxacin and vancomycin, and Pseud. aeruginosa sensitivity to piperacillin, amikacin, tobramycin was generally good. E. coli showed a satisfactory susceptibility on average, and P. mirabilis showed a good sensitivity to piperacillin, cephalosporins, amikacin, tobramycin, aztreonam and imipenem. Thus, the general bacterial flora and susceptibility have remained mostly unchanged over the years, with the conspicuous exception of Enterococcus spp. and E. cloacae, which demonstrated a marked increase in incidence, with a concomitant dramatic decrease in the sensitivity of Enterococcus spp. to antibiotics.     

Effect of Surface Characteristics on Compressive Stress and Leakage Rate in Gasketed Flanged Joints

The effect of surface characteristics on compressive stress and leakage rate in gasketed flanged joints is discussed qualitatively and quantitatively based on experimental verifications. The sensitivity of the sealing phenomenon to the sealing surface characteristics, gas pressure and stress level is presented. Sealing surfaces produced with grinding, turning and milling procedures of different roughness values were the subject of the tests with three types of gasket, namely, PTFE (polytetrafluoroethylene), graphite, and spiral wound. The experimental results indicated that the roughness value Ra (average arithmetic height) has no effect on the sealing performance of the gasket, except for the ground type sealing surfaces. The platen (flange) surface forms were found to be the determining factor on the leakage rate and flow regime for gasketed flanged joints. It is shown that the surface characteristics have no effect on the leakage rate and on the gasket performance for the PTFE gasket at high stress levels. However, at low stress levels the rougher ground and milled sealing surfaces having radial channels on the surface produce larger leakage rates. The effect of surface characteristics, stress level, and gas pressure was more evident on the graphite and spiral wound gasket types, at both low and high stress levels. It is also shown that the leakage rate follows an exponential law as a factor of surface stress and gas pressure, for all sealing surface characteristics and gasket types. While there is little difference in the leakage rate for the PTFE gasket when the gasket stress level is increased by a factor of two, from S3 to S5, a substantial leakage rate reduction by a ratio of 17.5 times was observed for the graphite and spiral wound gaskets for most sealing surfaces. On the other hand, when increasing the helium gas pressure from 200 to 800 p.s.i., the leakage rate increased by 5 times for the PTFE, and up to10 times for the graphite and the spiral wound gaskets. It was determined that while molecular flow regimes can be achieved for the PTFE gasket for all sealing surfaces, the two other gasket types provided laminar flow under the same operating conditions.     

PALAEOZOIC TECTONIC HISTORY OF SE TURKEY

A review of published data for the Palaeozoic of SE Turkey, together with facies distribution and palaeo-fault maps derived from regional field data, are used to interpret the Palaeozoic tectonic history of the region.
Intracratonic rifting events in the Early Cambrian and the Early Ordovician led to syn-rift deposition within fault-bounded basins. Marine transgressions across the region in the mid-Cambrian and mid-to-Late Ordovician were probably influenced by regional thermal subsidence after each rifting event, in addition to (glacio-)eustatic mechanisms. Marine transgressions in the Early Silurian and Early Carboniferous are thought to have been entirely eustatic in origin.
Poorly-constrained, Late Palaeozoic facies variations across the region can be related to uplift during the Caledonian and Hercynian orogenic episodes. Widespread tilting and erosion before the Cretaceous, which has removed much of the Late Palaeozoic record from this region, resulted from uplift at the edges of a major rift system which was initiated during the Triassic-to-Jurassic opening of the Southern Neotethys.     

Nuclear Magnetic Resonance Studies of Silicon Carbide Polytypes

The ²⁹Si MAS NMR spectra of the 2H, 4H, 6H, and 3C polytypes of silicon carbide are presented. An attempt is made to correlate differences in the chemical shifts with local atomic environment. The results of the analysis of the spectra of pure polytypes are used as a basic for the interpretation of the spectra of mixed polytypes and a discussion of the crystallinity and impurity levels of different samples. Carbon-13 chemical shifts obtained from spectra of the same polytypes are also tabulated.