Mutational analysis of structure--activity relationships in human tumor necrosis factor-alpha

To determine the region of human tumor necrosis factor-alpha(TNF-), essential for cytotoxic activity against mouse L-M cells,single amino-acid-substituted TNF- mutant proteins (muteins)were produced in Escherichia coli by protein engineering techniques.An expression plasmid for TNF- was mutagenized by passage throughan E.coli mutD5 mutator strain and by oligonucleotide-directedmutagenesis. Approximately 100 single amino-acid-substitutedTNF- muteins were produced and assayed for cytotoxic activity.The cytotoxic activities of purified TNF- muteins, e.g. TNF-31T,-32Y, -82D, -85H, -115L, -141Y, -144K and -146E, were < 1%of that of parent TNF-. These results indicate that the integrityof at least four distinct regions of the TNF- molecule is requiredfor full biological activity. These regions are designated asfollows: region I, from position 30 to 32; region II, from position82 to 89; region III, from position 115 to 117; region FV, fromposition 141 to 146. In addition, TNF-141Y could not completelycompete with parent TNF- for binding to the receptor. This demonstratesthat region IV, and at least aspartk acid at position 141, mustbe involved in the TNF receptor binding site.     

Method for correcting molecular weight from gel-permeation chromatography. I. Skewing and concentration corrections

The purpose of this paper is to present a metod to calculate real molecular weight averages of polymer samples from GPC chromatograms where the instrumental spreading functions are skewed and the concentration effect exists. In this method, it is assumed that (1) the skewed chromatograms of monodisperse polymer samples can be represented as resultant of halves of two different Gaussians, (2) the resolution factors are regarded as constant in the case of low sample loading, and (3) the peak elution volume is independent of the presence of other components in the case of low sample loading. Adequate monodisperse polystyrenes and the mixtures (binary, seven and ten components) were examined for this purpose; and the molecular weight averages calculated by this method were compared with the ones obtained by Rosen and Provder's method. From the results in our study, it is found that this method can be available for correcting molecular weight from GPC chromatograms except for very narrow high molecular weight samples.     

Correlations of Hepatic Hemodynamics,Liver Function,and Fibrosis Markers in Nonalcoholic Fatty Liver Disease: Comparison with Chronic Hepatitis Related to Hepatitis C Virus

The progression of chronic liver disease differs by etiology. The aim of this study was to elucidate the difference in disease progression between chronic hepatitis C (CHC) and nonalcoholic fatty liver disease (NAFLD) by means of fibrosis markers, liver function, and hepatic tissue blood flow (TBF). Xenon computed tomography (Xe-CT) was performed in 139 patients with NAFLD and 152 patients with CHC (including liver cirrhosis (LC)). The cutoff values for fibrosis markers were compared between NAFLD and CHC, and correlations between hepatic TBF and liver function tests were examined at each fibrosis stage. The cutoff values for detection of the advanced fibrosis stage were lower in NAFLD than in CHC. Although portal venous TBF (PVTBF) correlated with liver function tests, PVTBF in initial LC caused by nonalcoholic steatohepatitis (NASH-LC) was significantly lower than that in hepatitis C virus (C-LC) (p = 0.014). Conversely, the liver function tests in NASH-LC were higher than those in C-LC (p < 0.05). It is important to recognize the difference between NAFLD and CHC. We concluded that changes in hepatic blood flow occurred during the earliest stage of hepatic fibrosis in patients with NAFLD; therefore, patients with NAFLD need to be followed carefully.     

Dissolution mechanism of cellulose in SO2–amine–dimethylsulfoxide

The dissolution mechanism of cellulose in SO₂–amine–dimethylsulfoxide systems was studied by using ¹H- and ¹³C-NMR spectroscopy. SO₂ and amine (diethylamine or triethylamine) were found to form a complex in DMSO, and the SO₂–amine complex, in turn, reacts with an alcoholic hydroxyl group of methanol to produce a new complex. In the case of cellulose, it was proved that all hydroxyl groups in cellulose react with the SO₂–amine complexes and form the same complexes in the solution state as those formed in methanol.     

Surface-Chemical Criteria for Optimum Adhesion

According to Bikerman, who attributes failure in adhints to a weak boundary layer, it is almost impossible and meaningless to correlate adhesive strength to surface-chemical properties of adhints. Though his assertion seems to be confirmed by the recent studies of Schonhorn and his coworkers on the methods of CASING and TCR, not a few results have yet been accumulated, which show a close relation between them. In this paper surface-chemical criteria for the optimum adhesion are investigated and the minimum interfacial tension or the maximum wetting pressure is deduced from the published data and our own as a first approximation. It is emphasized that, when critical surface tension γ_c would be used as a measure of surface-chemical properties of solid, its variability according to liquid series (nonpolar, polar and hydrogen bonding liquids) should be carefully taken into consideration. The importance is shown for polyethylene and its fluorine substituted polymers, using newly measured contact angle data and Zisman's data. Results of Levine et al. and Schonhorn et al. on adhesive shear strength with epoxy adhesives are replotted against available values of γ_c obtained by the use of hydrogen bonding liquid (γ_c^c), which are thought to reflect wetting behaviors of epoxy adhesives quite well. Each curve shows a maximum around γ_c^c = 40 dyne/cm with few points falling off the curves.     

Degradation of polymers by high-speed stirring

Benzene solutions containing either poly(ethylene oxide) or poly(methyl methacrylate) or both of varying concentrations were prepared, stirred at a speed of 30,000 rpm, and the effects of concentrations of the same species or different species of polymers on the degradation of the polymers were investigated. Within experimental error little difference was observed in the [η] curves measured for each degraded polymer in spite of change in concentration. It is clear that the degradation of polymer with high-speed stirring in the concentration range 1–4% w/v is not caused by the interaction of polymer chains.     

Prism-pair interferometry by homodyne interferometers with a common light source for high-accuracy measurement of the absolute refractive index of glasses

A prism-pair interferometer comprising two homodyne interferometers with a common light source was developed for high-precision measurements of the refractive index of optical glasses with an uncertainty of the order of 10(-6). The two interferometers measure changes in the optical path length in the glass sample and in air, respectively. Uncertainties in the absolute wavelength of the common light source are cancelled out by calculating a ratio between the results from the interferometers. Uncertainties in phase measurement are suppressed by a quadrature detection system. The combined standard uncertainty of the developed system is evaluated as 1.1×10(-6).     

Stabilization effect of Co for Mo phase in CoMoAl2O3 hydrodesulfurization catalysts studied with X-Ray photoelectron spectroscopy

The promotional effects of Co in $\text{CoMo}\text{Al}{}_2\text{O}{}_3$ hydrodesulfurization (HDS) catalysts were studied by means of X-ray photoelectron spectroscopy. The higher MoO₃-content $\text{Mo}\text{Al}{}_2\text{O}{}_3$ catalysts (10 and 20 wt% MoO₃) contain mobile Mo, which migrates from the pores to the outermost surface layers of the catalysts and segregates to form less active crystalline MoS₂ during the HDS reaction, while in the case of $\text{Mo}\text{Al}{}_2\text{O}{}_3$ (5 wt% MoO₃) catalyst: no migration of Mo was observed. It is revealed that the Co in $\text{CoMo}\text{Al}{}_2\text{O}{}_3$ catalyst inhibits the migration and segregation of Mo and that it keeps Mo effective for the HDS reaction, since no surface enrichment of Mo was observed. It is concluded that stabilization of the Mo monomolecular layer is the main role of Co. The active species of Mo is suggested to have the composition of S/Mo(IV) = 1 on the basis of the sulfur contents of the catalysts under the mild HDS reaction conditions.     

Comparative hypocholesterolemic effects of six vegetable oils in cholesterol-fed rat

The hypocholesterolemic efficacies of various polyunsaturated fatty acids were compared in rats given cholesterol-enriched diets.Oenothera biennis Linn oil (OBLO, linoleic +γ-linolenic), sunflower oil (linoleic), palm oil (PLO, oleic+linoleic), soybean oil (linoleic+α-linolenic), high-oleic safflower oil (oleic+linoleic), or mixed oil (linoleic+α-linolenic) was added to the diet at 200 g/kg (20% groups). OBLO was also added at 100 g/kg diet (10% group). The serum total and very low density lipoprotein+intermediate lipoprotein+low density lipoprotein cholesterol concentrations of the 10 and 20% OBLO groups were consistently lower than those in the other groups. The liver cholesterol concentration in the PLO group was lower in all groups. The liver cholesterol concentrations in the 10 and 20% OBLO groups were also lower than in the other groups. There were no significant differences in the fecal neutral sterol and bile acid extraction among groups.     

Hydrodesulfurization activity of highly dispersed Co sulfide clusters prepared in zeolite cages

Zeolite-supported highly dispersed Co sulfide clusters are synthesized using Co(CO)₃ (NO) as a precursor. The amount of Co inCoS_x/zeolite anchored by a CVD technique increases as the Al/Si ratio of the zeolite increases, whereas the activity per Co atomdecreases for thiophene hydrodesulfurization (HDS). When an Na-exchanged USY zeolite is used, the Co sulfide catalyst shows a muchhigher HDS activity than a conventional Co—Mo/Al₂ O₃ catalyst. It is considered from XAFS and NO adsorption techniques that thehigh HDS activity of CoS_x/USY-Na is due to an extremely high dispersion of Co sulfide clusters. This revised version was published online in July 2006 with corrections to the Cover Date.