Molecular phylogenetic characterization of Streptomyces protease inhibitor family

We previously found that proteinaceous protease inhibitors homologous to Streptomyces subtilisin inhibitor (SSI) are widely produced by various Streptomyces species, and we designated them "SSI-like proteins" (Taguchi S, Kikuchi H, Suzuki M, Kojima S, Terabe M, Miura K, Nakase T, Momose H [1993] Appl Environ Microbiol 59:4338-4341). In this study, SSI-like proteins from five strains of the genus Streptoverticillium were purified and sequenced, and molecular phylogenetic trees were constructed on the basis of the determined amino acid sequences together with those determined previously for Streptomyces species. The phylogenetic trees showed that SSI-like proteins from Streptoverticillium species are phylogenetically included in Streptomyces SSI-like proteins but form a monophyletic group as a distinct lineage within the Streptomyces proteins. This provides an alternative phylogenetic framework to the previous one based on partial small ribosomal RNA sequences, and it may indicate that the phylogenetic affiliation of the genus Streptoverticillium should be revised. The phylogenetic trees also suggested that SSI-like proteins possessing arginine or methionine at the P1 site, the major reactive center site toward target proteases, arose multiple times on independent lineages from ancestral proteins possessing lysine at the P1 site. Most of the codon changes at the P1 site inferred to have occurred during the evolution of SSI-like proteins are consistent with those inferred from the extremely high G + C content of Streptomyces genomes. The inferred minimum number of amino acid replacements at the P1 site was nearly equal to the average number for all the variable sites. It thus appears that positive Darwinian selection, which has been postulated to account for accelerated rates of amino acid replacement at the major reaction center site of mammalian protease inhibitors, may not have dictated the evolution of the bacterial SSI-like proteins.     

Living cationic polymerization of isobutyl vinyl ether initiated by the trimethylsilyl iodide/zinc iodide system

Summary Trimethylsilyl iodide in conjunction with zinc iodide (Me₃SiI/ZnI₂) as an initiating system led to living cationic polymerization of isobutyl vinyl ether in toluene at 0 or –40°C or in methylene chloride at –40°C (ZnI₂ was dissolved in acetone). The number-average molecular weight of the polymers was directly proportional to monomer conversion and in excellent agreement with the calculated value assuming that one polymer chain forms per unit trimethylsilyl iodide. At room temperature (+25°C), however, the polymerization failed to give perfectly living polymers; the polymer molecular weight was smaller than the calculated value. On addition of a fresh feed of monomer at the end of the polymerization at –40°C, the added feed was smoothly polymerized at nearly the same rate as in the first stage, and the polymer molecular weight continued to increase in direct proportion to monomer conversion. Throughout the reaction, the molecular weight distribution of the polymers stayed very narrow (M_w/M_n< 1.1).Living cationic polymerization of vinyl ethers by electrophile/Lewis acid initiating systems, part 2. For part 1 see ref. 2     

Catheter manipulation training system based on quantitative measurement of catheter insertion and rotation

How to preserve the catheter without any modifications and improve the precision of manipulation in the catheter-based endovascular intervention remains a challenge for quantitative training and evaluation. This paper presents an optical sensor unit to measure catheter motions based on the working principle of Laser Doppler, which allows for catheter motion measurement with non-contact detection and supports medical training and evaluation without modifications on the catheter. Then, calibrate this sensor unit and implement recording of catheter motion. Next, propose a training system by tracking the reference trajectory based on an expert’s skill, and provide visual feedback to improve the precision of catheter manipulation. Eight subjects were trained for catheter manipulation with this system. Among them, six subjects improved their catheter manipulation of linear motion, and seven improved the catheter rotational motion.     

Rheological and morphological characterization of selected poly(organo)phosphazenes

Rheological properties of selected poly(organo)phosphazenes have been studied using capillary rheometry. Melt viscosities of polymers were calculated as a function of shear stress and shear rate. Thermal stability was also studied. In the range of shear rate studied, all polyphosphazenes showed shear thinning behavior. The power law model was fitted to the data to get the power law index and the consistency index. Poly[bis(phenoxy)phosphazene] showed a Newtonian plateau at low shear rates whereas poly[bis(trifluoroethoxy)phosphazene] and poly[bis(p-phenylphenoxy)phosphazene] which exhibited liquid crystalline character were shear thinning even at low shear rates. Solid state capillary extrusion of polyphosphazenes was found to be more successful in fiber formation than other “state-of-the-art” processes, such as gel-spinning. The molecular weights of all polyphosphazenes synthesized were found to be too low for, Couette fiber spinning. Chemical, physical, and morphological studies were made on all fibers prepared. Many investigative techniques, such as X-ray diffraction, differential scanning calorimetry (DSC), density measurements, scanning electron microscopy (SEM), optical microscopy, solution nuclear magnetic resonance (NMR) spectroscopy, and gel permeation chromatography (GPC) analysis are featured in this paper as characterization tools.     

Interaction of nylon 6-clay surface and mechanical properties of nylon 6-clay hybrid

We synthesized nylon 6-clay hybrid materials using four types of clay minerals, montmorillonite, saponite, hectrite, and synthetic mica. The mechanical properties of their injection molded specimens were measured according to ASTM. Nylon 6-clay hybrid using montmorillonite was superior to the other hybrids in mechanical properties. This might result from the difference in the interaction between nylon molecules and silicates in the hybrids. To clarify this hypothesis, we synthesized intercalated compounds of the clay minerals with glycine as the model of the hybrids, and analyzed the interaction using ¹⁵N cross polarization magic angle spinning (CP/MAS) NMR spectroscopy. The ¹⁵N-NMR result reveals that the positive charge density on the nitrogen of the intercalated compound based on montmorillonite was largest in all the intercalated compounds. It was suggested that montmorillonite interacted strongly with nylon 6 by ionic interaction. This ionic interaction was one of the reasons why these hybrid materials had superior mechanical properties. © 1995 John Wiley & Sons, Inc.     

Enzyme-resistant fractions of beans lowered serum cholesterol and increased sterol excretions and hepatic mRNA levels in rats

Feeding rats beans with resistant starch reduces their serum cholesterol concentration; however, the mechanism by which this occurs is not fully understood. We examined the effects of enzyme-resistant fractions of adzuki (Vigna angularis) and tebou (Phaseolus vulgaris, var.) beans on serum cholesterol and hepatic mRNA in rats. Rats were fed a cholesterol-free diet with 50 g of cellulose powder (CP)/kg, 50 g of an enzyme-resistant fraction of adzuki starch (AS)/kg, or 50 g of an enzyme-resistant fraction of tebou starch (TS)/kg diet for 4 wk. There were no significant differences in body weight, liver weight, and cecum contents among the groups, nor was there a significant difference in food intake among the groups. The levels of serum total cholesterol, VLDL + intermediate density lipoprotein + LDL-cholesterol, and HDL cholesterol in the AS and TS groups were significantly (P<0.05) lower than in the CP group throughout the feeding period. Total hepatic cholesterol in the CP group was significantly (P<0.05) lower than in the AS and TS groups, fecal cholesterol excretion in the TS group was significantly (P<0.05) greater than in the CP and AS groups, and the fecal total bile acid concentrations in the AS and TS groups were significantly (P<0.05) higher than in the CP group. Cecal acetate, propionate, and n-butyrate concentrations in the AS and TS groups were significantly (P<0.05) higher than in the CP group. The level of hepatic scavenger receptor class B1 (SR-B1) mRNA in the TS group was significantly (P<0.05) higher than in the CP group, and the levels of hepatic cholesterol 7α-hydroxylase mRNA in the AS and TS groups were significantly (P<0.05) higher than in the CP group. These results suggest that AS and TS have a serum cholesterol-lowering function due to the enhanced levels of hepatic SR-B1 and cholesterol 7α-hydroxylase mRNA.     

Fluorescence property of polycarbonate related to radial‐pattern formation

The pattern change in a laminar flow of molten polycarbonate in a disk mold was experimentally investigated by altering the flow velocity. By use of a light‐scattering method, a radial pattern characterized by a periodic array of needlelike regions and the matrix was found to form as the flow velocity was increased. The density distribution of the polycarbonate molecules was an essential factor for pattern formation. As a result of the formation of this pattern, it was found that the boundaries between the needlelike regions and the matrix emitted weak fluorescence in the wavelength range of 600–700 nm under laser irradiation. An analysis indicated that transitions between excited π electron states in the phenylene groups were the sources of this fluorescence and that a key factor behind its occurrence was a decrease in the molecule density at the boundaries. These results reveal that the radial‐pattern formation changes the fluorescence property of polycarbonate in the visible and near‐infrared regions. © 2004 Wiley Periodicals, Inc. J Appl Polym Sci 92: 468–473, 2004     

Promotion effects of Pt and Rh on catalytic performances of Mo/HZSM-5 and Mo/HMCM-22 in selective methane-to-benzene reaction

The promotion effects of Pt and Rh on catalytic performances of Mo/HZSM-5 and Mo/HMCM-22 in selective methane-to-benzene reaction were studied in the presence of additive H₂. The selectivity to naphthalene was effectively suppressed and highly selective and stable benzene formation was obtained by the addition of noble metal to the Mo/HZSM-5 and Mo/HMCM-22 catalysts, due to the suppression of carbon deposition on the Brønsted acid sites of zeolite.     

Thiobarbituric acid-reactive substances from peroxidized lipids

The thiobarbituric acid (TBA) reaction was performed on linoleic acid 13-monohydroperoxide, autoxidized fatty esters, edible fats and oils, rat liver microsomal lipids, and on human erythrocyte ghost lipids in order to determine which substances from peroxidized lipids are TBA-reactive. The reaction was carried out in 2% acetic acid containing butylated hydroxytoluene using two different reaction modes: a one-step mode which involves heating at 100°C, and a two-step mode which involves first treatment at 5°C and subsequent heating at 100°C. Yields of the red 1∶2 malonaldehyde/TBA adduct, as estimated by absorbance, fluorescence intensity and high-performance liquid chromatography, were much higher than the malonaldehyde content as determined by direct chemical analysis. Yields of red pigment obtained by the two-step mode were slightly higher than those obtained by the one-step mode. Pigment yields were dramatically increased by addition oft-butyl hydroperoxide. Red pigment formation from alkenals and alkadienals was similarly enhanced by the two-step mode or by addition oft-butyl hydroperoxide, whereas pigment formation from malonaldehyde was not. It appears likely that a component of the total red pigment formed from the peroxidized lipids was due to aldehyde species other than malonaldehyde.     

Solution properties of xanthan. Light scattering and viscometry on dilute and moderately concentrated solutions

Elastic and quasi-elastic light-scattering, viscometric and rheological studies are given for solutions of the microbial polysaccharide Xanthomonas campestris (xanthan) in aqueous 0.62N NaCl for polymer concentrations from 0.03 to 2.2 g kg⁻¹. The observed negative ∂ln[η]/∂ln T is interpreted as a decrease of the persistence length with increasing T. The behaviour in moderately concentrated solutions (2<[η]c<25) reveals intermolecular association, leading to gel formation in the extreme case. The effect of the association on the viscometric and light-scattering data is discussed. It is concluded that the early stages of association involve structure with the chain axes nearly parallel, but that larger, particulate-like structures develop with increasing c, eventually leading to gel formation under certain conditions.