Novel sugar phosphotransferase system applicable to the efficient labeling of the compounds synthesized via the non-mevalonate pathway in Escherichia coli

It is known that 1-deoxy-D-xylulose taken up by Escherichia coli is used as the precursor of the compounds synthesized via the non-mevalonate pathway, such as isoprenoids, probably after conversion into 1-deoxy-D-xylulose 5-phosphate. In this report, we show that a novel phospho(enol) pyruvate-dependent phosphotransferase system catalyzes the uptake and phosphorylation of 1-deoxy-D-xylulose.     

Deletion and insertion of a 192-residue peptide in the active-site domain of glycosyl hydrolase family-2 beta-galactosidases

The monomeric multimetal-binding beta-galactosidase of Saccharopolyspora rectivirgula (srbg), a glycosyl hydrolase family-2 enzyme, has a unique sequence consisting of 192 amino acid residues with no similarity to known proteins. This 192-residue sequence (termed the "iota [iota] sequence") appears to be inserted into a sequence homologous to the active-site domain of the Escherichia coli lacZ enzyme (lacZbg). To assess the effects of the t sequence at specific sites of beta-galactosidase on the catalytic functioning and molecular properties of beta-galactosidase, deletion or insertion mutants of beta-galactosidases were constructed, expressed in LacZ- E. coli strains, and characterized: srbgdelta in which the iota sequence was deleted from srbg, and lacZbgI, in which the 192-residue iota sequence was inserted into the corresponding position (between Asp591 and Phe592) in the active-site domain of lacZbg. srbgdelta was a catalytically inactive, dimeric protein which retained multimetal-binding characteristics, suggesting that the iota sequence is very important for maintaining the structure necessary for the catalytic functioning and the monomeric structure of srbg but is not responsible for the unique metal ion requirements of srbg. On the other hand, lacZbgI existed as a mixture of a monomer, a tetramer, and higher multimers. The monomeric species was inactive, whereas the tetramer and other multimers were catalytically active (V(max )K(m) value, 25% of that of lacZbg) and highly specific for beta-D-galactoside. The tetrameric lacZbgI was activated by Mg2+ and Mn2+ with lowered metal affinities, and the stoichiometry of metal binding was unchanged from that of lacZbg. These results, along with the published stereo structure of lacZbg, suggest that, in lacZbgI, the inserted 192-residue iota peptide could fold independently of the lacZbg domains into a "sub-domain," lying distant from the active site and subunit interfaces.     

Dynamic moduli for short fiber-CR composites

The dynamic moduli, E′ and E″, and tan δ for nylon–CR and PET–CR composites with unidirectional short fibers were studied as a function of temperature by using a Rheovibron. The temperature dependence of tan δ showed two dispersion peaks for nylon–CR composite. The peak at ?28°C corresponded to the main dispersion of CR and the peak at 100°C to the α-dispersion of nylon 6. For a PET-CR composite, in addition to the individual dispersion of CR and PET, a small and broad peak was observed at about 90°C. The angular dependence of E′ indicated that the short fibers assumed good orientation. The storage modulus for the composites was given by the parallel model as E′ = V_f′E_f + V_mE′_m., where E′_c, E′_f and E′_m were the storage modulus for the composite, fiber, and matrix and V_f and V_m were the volume fraction of fiber and matrix, respectively. In the transverse direction of fiber, the peak values of tan δ at ?28°C were given by the following equation; tan δ_c = tan δ_m ? δV_f, where tan δ_c and tan δ_m are the loss tangent for the composite and matrix, respectively, and α is coefficient depending on fiber type. The results indicated that a region with strong interaction was formed between fibers and CR matrix.     

Simple determination of residual anticoccidial drugs (diclazuril and nicarbazin) in chicken tissues by HPLC

A simple and rapid determination of anticoccidial drug residues, diclazuril (DCZ) and nicarbazin (NCZ), in chicken tissues has been developed. DCZ and NCZ were extracted with acetonitrile from chicken liver, muscle, and fat. The extract was rinsed with n-hexane saturated with acetonitrile and then evaporated. The residue was dissolved in 1.4 mL of acetonitrile-methanol (1:1), then 1.0 mL of n-hexane saturated with acetonitrile-methanol (1:1) was added, and the mixture was partitioned by the addition of 0.6 mL of water. DCZ and NCZ in the aqueous layers were determined by HPLC on an Xterra RP-18 column with acetonitrile-0.5% ammonium acetate containing 0.01 mol/L tetra-n-butylammonium hydrogen sulfate (43:57) as the mobile phase. The mean recoveries (n = 5) of DCZ and NCZ spiked in chicken tissues at the maximum residue levels were 92.0-95.6% (CV 2.4-3.0%) and 87.3-89.4% (CV 1.7-2.8%), respectively. The detection limits of DCZ and NCZ were 0.01 and 0.004 microgram/g, respectively.     

Bioconversion of 1-adamantanol to 1,3-adamantanediol using Streptomyces sp. SA8 oxidation system

To efficiently produce 1,3-adamantanediol (1,3-ad(OH)₂) from 1-adamantanol (1-adOH), our stocks of culture strains and soil microorganisms were surveyed for hydroxylation activity towards 1-adOH. Among them, the soil actinomycete SA8 showing the highest hydroxylation activity was identified as Streptomyces sp. based on 16S ribosomal DNA sequence analysis. The reaction products were purified by silica gel column chromatography, and from NMR and MS analyses, they were identified as 1,3-ad(OH)₂ and 1,4-ad(OH)₂. Streptomyces sp. SA8 produced 5.9 g l^? 1 1,3-ad(OH)₂from 6.2 g l^? 1 1-adOH in culture broth after 120 h at 25 °C. Using resting cells, 2.3 g l^? 1 1,3-ad(OH)₂ was produced after 96 h of incubation at a 69% conversion rate. In both cases, 1,4-ad(OH)₂ was formed as a byproduct at a rate of about 15%. Strain SA8 also hydroxylated 2-adamantanol and 2-methyl-2-adamantanol.     

Multiresidue determination of quinolones in animal and fishery products by HPLC

A simple and rapid multiresidue method was developed for the determination of twelve quinolones (ciprofloxacin, danofloxacin, difloxacin, enrofloxacin, flumequine, marbofloxacin, nalidixic acid, norfloxacin, ofloxacin, orbifloxacin, oxolinic acid and sarafloxacin) in muscle, liver, chicken eggs, milk, prawn and rainbow trout.The quinolones were extracted from a sample with acetonitrile-water (95 : 5). A fifth part of the filtered extract was diluted with water to keep the acetonitrile ratio at ca. 60%, and passed through a C18 mini-column. The eluate was evaporated to dryness, and the residues were dissolved in methanol-water (30 : 70) for HPLC analysis.The quinolones were separated on a Inertsil ODS-3V column (4.6 mm i.d.x250 mm) with a gradient system of 0.1% phosphoric acid-acetonitrile as the mobile phase, with fluorescence detection.No interfering peak was found on the chromatograms of animal and fishery products, except for milk. The recoveries of the quinolones were over 60% from the animal and fishery products fortified at 0.1 microg/g, and the quantification limits of the quinolones were 0.005 microg/g. This proposed method was found to be effective and suitable for the screening of the quinolones in animal and fishery products.     

Multilayered reasoning by means of conceptual fuzzy sets

The real world consists of instances of events and continuous numeric values, while people represent and process their knowledge in terms of symbols. Fuzzy sets provide a strong notation connecting the symbolic representation to the real world. In previously proposed Conceptual Fuzzy Sets (CFS), the meaning of a concept is represented by the distribution of activations of labels in a bidirectional associative memory. In particular, a multilayered structured CFS represents the meaning of the same concept as it is used in various expressions in each layer. The propagation of activations corresponds to reasoning. Therefore, we propose a multilayered reasoning method associated to a multilayered structured CFS, which has the following features: (1) capable of simultaneous symbolic and quantitative processing, (2) capable of simultaneous top-down and bottom-up processing. The effectiveness of the proposed method is illustrated by practical examples of decision regarding the amount of steering in the task of parking a car, and recognition of facial expressions for an image understanding system. © 1996 John Wiley & Sons, Inc.     

Development of a new heterojunction structure (ACJ -HIT) and its application to polycrystalline silicon solar cells

A new solar cell structure named HIT (Heterojunction with Intrinsic Thin layer) has been developed based on new artificially constructed junction (ACJ) technology. In this structure a non-doped a-Si thin layer was inserted between the p(a-Si)/n(c-Si) heterojunction, improving the output characteristics and achieving a conversion efficiency of 18.1%. This structure was applied to cast polycrystalline silicon solar cells of a practical size. A high conversion efficeincy of 13.6% was obtained with a cell size of 10 cm × 10 cm using various technologies, including hydrogen plasma passivation.     

Photoacoustic Spectroscopy of Some Energetic Materials

To find an effective laser source to ignite energetic materials, the absorption spectra of some energetic materials are obtained by means of a photoacoustic spectroscopy (PAS). In this experiment, PAS covers the wavelength region of 400 nm-1600 nm in which no other conventional method can take absorption spectra for powdered energetic materials. Photoacoustic spectra of 18 energetic materials are reported. In general, energetic materials tested showed peaks in 600 nm–800 nm and 1400 nm–1600 nm ranges. It is found that the energy required to initiate explosives in the case of ruby laser initiation were correlated with their photoacoustic signal intensities.