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.     

An investigation of the pH-activity relationships of Cex, a family 10 xylanase from Cellulomonas fimi: xylan inhibition and the influence of nitro-substituted aryl-beta-D-xylobiosides on xylanase activity

The kinetic parameters of Cex, a family 10 xylanase from Cellulomonas fimi, were determined at various pH levels using soluble birchwood xylan (BWX) as a natural polymeric substrate along with three other synthetic aryl-beta-D-xylobioside substrates. Using BWX, a high level of substrate inhibition was observed which increased with decreasing pH. In contrast, typical Michaelis-Menten-type profiles were obtained using the three aryl-beta-D-xylobiosides as substrates. The k(cat) values determined using o-nitrophenyl-beta-D-xylobioside did not change as the pH increased, whereas the k(cat) values obtained with BWX, phenyl-beta-D-xylobioside and p-nitrophenyl-beta-D-xylobioside decreased, suggesting that the presence of an ortho nitro group affects the activity displayed by Cex. These differences were not observed with XynB from Clostridium stercorarium F9, a member of the same family of xylanases as Cex. These results indicate that a careful evaluation is required when employing substituted aryl-beta-D-xylobiosides in the characterization of xylanases.     

Interactive segmentation of the cerebral lobes with fuzzy inference in 3T MR images.

Measurement of volume and surface area of the frontal, parietal, temporal and occipital lobes from magnetic resonance (MR) images shows promise as a method for use in diagnosis of dementia. This article presents a novel computer-aided system for automatically segmenting the cerebral lobes from 3T human brain MR images. Until now, the anatomical definition of cerebral lobes on the cerebral cortex is somewhat vague for use in automatic delineation of boundary lines, and there is no definition of cerebral lobes in the interior of the cerebrum. Therefore, we have developed a new method for defining cerebral lobes on the cerebral cortex and in the interior of the cerebrum. The proposed method determines the boundaries between the lobes by deforming initial surfaces. The initial surfaces are automatically determined based on user-given landmarks. They are smoothed and deformed so that the deforming boundaries run along the hourglass portion of the three-dimensional shape of the cerebrum with fuzzy rule-based active contour and surface models. The cerebrum is divided into the cerebral lobes according to the boundaries determined using this method. The reproducibility of our system with a given subject was assessed by examining the variability of volume and surface area in three healthy subjects, with measurements performed by three beginners and one expert user. The experimental results show that our system segments the cerebral lobes with high reproducibility.     

Crystallographic Orientation Dependence of the Schottky Properties of Au/SrTiO3 Junctions

Crystallographic orientation dependence of the Schottky properties of Au/Nb-doped SrTiO₃ (STO:Nb) junctions has been investigated using single crystals of STO:Nb (1 0 0) and (1 1 1). It is found from electrical properties that the Schottky barrier height (SBH) of the Au/STO:Nb junctions estimated from current density (J)-voltage (V) characteristics shows crystallographic orientation dependence, while the flat band voltage estimated from capacitance (C)-voltage (V) characteristics is independent of the orientation. Displacement currents originated from the junction capacitance have been clearly observed at reverse bias voltage even in a condition of dV/dt 8.75 × 10^–3 [V/s] because of large electrostatic permittivity of the STO, and the displacement currents also showed crystallographic orientation dependence. The different response in the electrical properties of the Schottky junctions suggests that electric properties of intrinsic low permittivity layers, which exist at Au/STO:Nb interfaces, have the crystallographic orientation dependence.     

c-axis Superfluid Response and Quasiparticle Conductivity in Bi2Sr2CaCu2O8+δ and Bi2Sr2CuO6+δ

Josephson plasma resonance for underdoped Bi ₂ Sr ₂ CaCu ₂ O ₈₊ and Bi ₂ Sr ₂ CuO ₆₊ have been measured by sweeping the microwave, frequency continuously. The resonance enables us to determine the superfluid density and quasiparticle conductivity in the c-axis accurately. We show that the. superfluid response and the low energy excitations out of the condensate in the c-axis of these materials are very different from those in the ordinary Josephson multilayer tunnel junctions. The Josephson coupling energy in single layer Bi ₂ Sr ₂ CuO ₆₊ is more than 5000 times smaller than is predicted in interlayer tunneling model.     

Space charge behavior in polymer film based on increment of capacitance under AC high field

Polymer materials have excellent dielectric and insulation properties; however, those properties in AC high field region have not been known well. Recently we established an evaluation method of high‐field AC dissipation current waveform of polymer materials 1 . AC dissipation current waveforms of polyethylene and polypropylene films show nonlinear distortion in AC high‐field region. This nonlinearity was thought to be related to the behavior of AC space charge formation in the sample near electrodes. The properties of space charge formed under AC high field at power frequency seem to differ from those formed under DC high field. The measurement of AC space charge distribution is not so easy due to the resolution limit of the space charge measurement. We studied the dielectric properties of biaxially oriented polypropylene (BOPP) film under AC high field up to 120 °C. It was found that tan δ, AC dissipation current (Ixr), and unbalanced component of capacitive current (ΔIxc) increased when the temperature became higher. In particular, ΔIxc increased above some threshold field and was considered to be due to the AC space charge formation. This AC space charge layer near electrode is thought to be formed due to carrier injection under AC high‐field application. Usually, the carrier mobility becomes smaller on lowering the temperature. Most of the carriers injected from the electrode are trapped near the electrode in the sample film. But in the high‐temperature region, the carrier mobility becomes larger and the carrier injection starts to increase from lower field. Many more carriers are injected from the electrode. It is thought that some of the injected carriers are trapped inside the sample film; the others go through the sample to the opposite side. © 2002 Wiley Periodicals, Inc. Electr Eng Jpn, 141(2): 8–16, 2002; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/eej.10018     

Chemorheology of an epoxy resin for pultrusion

The Williams-Landel-Ferry (W-L-F) equation was shown to describe the temperature dependence of the viscosity of a sulfonium accelerated, amine cured epoxy resin that had been formulated for pultrusion. The reference temperature and the viscosity at the reference temperature of the W-L-F equation were found to be simple functions of the resin advancement as determined by isothermal differential scanning calorimetry, A method is described of constructing the baseline for temperature-programmed differential scanning calorimetry analyses in which the heat capacity is changing due to reaction.     

Measurement and correlation of mutual diffusion coefficients for styrene-butadiene rubber(SBR)-n-nonane systems

The mutual diffusion coefficients for styrene-butadiene rubber(SBR)-n-nonane systems were measured using a sorption apparatus with a quartz spring at 100 and 130°C for mass fractions of n-nonane ranging from 0 to 0.16. The mutual diffusion coefficients were correlated with good agreement by the free-volume theory.     

Asymmetric oxidation of poly(vinyl sulfides) with percamphoric acid,aspergillus niger,and penicillium notatum

The asymmetric oxidation of poly(phenylvinyl sulfide) (poly-Ph VS) and poly(t-butylvinyl sulfide) (poly-t-BuVS) was carried out with optically active percamphoric acid, and the optically active polysulfoxides were obtained. The values of the specific rotation for the oxidation products of poly-PhVS and poly-t-BuVS were + 1.0 (having 56% of sulfoxide unit contents) and +9.1 (49%), respectively. The specific rotation of the product increased with increasing the contents of sulfoxide units. The optical rotatory dispersion curves of the oxidation products of poly-PhVS and poly-t-BuVS were positive curves and were found to fit the simple Drude equation. The λ_c values of the oxidation products (poly-PhVS, 271 mμ; poly-t-BuVS, 212 mμ) suggested that the chromophore which caused optically activity was the sulfoxide group. Similarly, the asymmetric oxidation of t-BuVS–MMA copolymer, t-BuVS–styrene copolymer, and PhVS–MMA copolymer was carried out, and the optically active copolysulfoxides were obtained. Furthermore, the biosynthetic oxidation of poly-PhVS and PhVS–maleic anhydride copolymer treated with aqueous KOH was carried out using Aspergillus niger or Penicillium notatum in Czapeck solution, and the optically active polymers were obtained.