Alteration of substrate specificity of cholesterol oxidase from Streptomyces sp. by site-directed mutagenesis

Despite the structural similarities between cholesterol oxidasefrom Streptomyces and that from Brevibacterium, both enzymesexhibit different characteristics, such as catalytic activity,optimum pH and temperature. In attempts to define the molecularbasis of differences in catalytic activity or stability, substitutionsat six amino acid residues were introduced into cholesteroloxidase using site-directed mutagenesis of its gene. The aminoacid substitutions chosen were based on structural comparisonsof cholesterol oxidases from Streptomyces and Brevibacterium.Seven mutant enzymes were constructed with the following aminoacid substitutions: L117P, L119A, L119F, V145Q, Q286R, P357Nand S379T. All the mutant enzymes exhibited activity with theexception of that with the L117P mutation. The resulting V145Qmutant enzyme has low activities for all substrates examinedand the S379T mutant enzyme showed markedly altered substratespecificity compared with the wild-type enzyme. To evaluatethe role of V145 and S379 residues in the reaction, mutantswith two additional substitutions in V145 and four in S379 wereconstructed. The mutant enzymes created by the replacement ofV145 by Asp and Glu had much lower catalytic efficiency forcholesterol and pregnenolone as substrates than the wild-typeenzyme. From previous studies and this study, the V145 residueseems to be important for the stability and substrate bindingof the cholesterol oxidase. In contrast, the catalytic efficiencies(k_cat/K_m) of the S379T mutant enzyme for cholesterol and pregnenolonewere 1.8- and 6.0-fold higher, respectively, than those of thewild-type enzyme. The enhanced catalytic efficiency of the S379Tmutant enzyme for pregnenolone was due to a slightly high k_catvalue and a low K_m value. These findings will provide severalideas for the design of more powerful enzymes that can be appliedto clinical determination of serum cholesterol levels and assterol probes.     

The effects of Ni and Li doping on the performance of lithium manganese oxide material for lithium secondary batteries

Layered Li_0.7[M_1/6Mn_5/6]O₂ (M=Li, Ni) was synthesized using a sol-gel method. P2-Na_0.7[M_1/6Mn_5/6]O₂ precursor was first synthesized by a sol-gel method, and then O2-Li_0.7[M_1/6Mn_5/6]O₂ was prepared by an ion exchange of Li for Na in P2-Na_0.7[M_1/6Mn_5/6]O₂ precursor. From charge/discharge curves, it was seen that Li_0.7[Li_1/6Mn_5/6]O₂ has two plateaus similar to those observed from a spinel structure, but Li_0.7[Ni_1/6Mn_5/6]O₂ holds a single plateau as observed from a typical layered structure. It was considered that Li_0.7[Li_1/6Mn_5/6]O₂ undergoes a phase transformation from layered to spinel structure during the charge/discharge cycle, but Li_0.7[Ni_1/6Mn_5/6]O₂ maintains O2-layered structure after the cycles. Li_0.7[Ni_1/6Mn_5/6]O₂ was higher in discharge capacity and retention rate than Li_0.7[Li_1/6Mn_5/6]O₂.     

Morphological study by TEM on uniaxially oriented thin films of PET, PEN and their blends

Uniaxially oriented thin films of poly(ethylene terephthalate) (PET), poly(ethylene 2,6-naphthalene dicarboxylate) (PEN) and their blends were prepared by applying shear strain to their respective melts, and the resulting morphologies were studied by transmission electron microscopy. Selected-area electron diffraction of each film revealed well-defined uniaxial orientation of polymer chains in the shearing direction. In the uniaxially oriented thin film of PEN, stacked-lamellar structure with the average long period of 27 nm consisting of a crystalline region about 15 nm thick and an amorphous one about 12 nm thick was found here and there in the dark-field image: PEN-type. On the other hand, stacked-lamellar structure was rarely observed in the case of PET: PET-type. In PET/PEN blends, the morphologies changed from the PET-type to the PEN-type with increasing content of PEN.     

A pre-heating vaporization technology of coal-water-slurry for the gasification process

A pre-heating vaporization technology of coal water slurry, CWS, was developed to increase the efficiency of coal gasification with the reduction of oxygen consumption. A CWS pre-heating experimental unit having a capacity of 2.0 tons-coal/day was manufactured. After the CWS was heated to 573 K at about 10 MPa, water in CWS was continuously vaporized in a tube of which the diameter increased gradually (2–10 mm I.D.), and dry coal particles with steam were atomized and fed to a vessel. No aggregation of coal particles was observed in the atomized samples. The temperature and the pressure distributions of the fluid were calculated with empirical heat transfer and pressure drop correlations. The calculated results well corresponded to the experimental data measured in some test points. Using the calculation model, a large scale apparatus of 100 tons-coal/day was designed using KNO₃ + NaNO₂ + NaNO₃ molten salt as a heating medium.     

Additional kernel observer: privilege escalation attack prevention mechanism focusing on system call privilege changes

International Journal of Information Security - Cyberattacks, especially attacks that exploit operating system vulnerabilities, have been increasing in recent years. In particular, if administrator...     

In-process evaluation of grit protrusion feature for fine diamond grinding wheel by means of electro-contact discharge dressing

This paper introduces a new in-process evaluation method for grit protrusion feature on wheel surface by monitoring discharge current trace during electro-contact discharge (ECD) dressing of metal-bonded fine diamond grinding wheel. First an impulse discharge machining experiment was carried out to investigate the correlation between metal bond removal and discharge parameters, namely discharge current I_e and discharge pulse duration τ_e. Then ECD dressing experiment for #600 diamond grinding wheel was conduced to analyses the quantitative effect of the discharge parameters (I_e and τ_e), derived from discharge current trace between wheel and dresser (electrode), on grit protrusion feature of wheel surface. The result shows that the grit protrusion feature is sensitive to the discharge parameters (I_e and τ_e) with reference to mean diamond grit size d_gm. Further, the discharge parameters (I_e and τ_e) in ECD dressing should conform to the discharge variables’ requirement of , by which the grit protrusion feature may be evaluated and the dressing process variables may be determined. Finally, the in-process evaluation method was successfully applied to ECD dressing of #1500 diamond grinding wheel for valid grinding of hard-brittle materials.     

金属结合剂#600金刚石砂轮的接触放电修锐实验研究

针对微细金刚石磨粒很难从金属结合剂砂轮的胎体中出刃的问题，开发出一种新修锐方法一接触放电修锐、它采用一种旋转复合电极修整轮与砂轮接触磨削，在金属结合剂胎体与电极间产生微小的脉冲放电，逐渐去除砂轮的金属结合剂，使微细金刚石在砂轮工作表面出刃，达到精密修锐的目的。通过对金属结合剂#600金刚石砂轮进行修锐实验，研究其有效性、修锐条件和实际应用。实验表明，该修锐方法不仅不损伤微细金刚石磨粒出刃刃角，而且还可以消除磨粒周围的结合剂尾状物，产生较好的容屑空间。磨削光学玻璃(BK10)的试验结果显示它比机械修锐能够更好地提高磨削表面质量，Ra达到0．12μm。     

A Nom historical document recognition system for digital archiving

A Nom historical document recognition system is being developed for digital archiving that uses image binarization, character segmentation, and character recognition. It incorporates two versions of off-line character recognition: one for automatic recognition of scanned and segmented character patterns (7660 categories) and the other for user handwritten input (32,695 categories). This separation is used since including less frequently appearing categories in automatic recognition increases the misrecognition rate without reliable statistics on the Nom language. Moreover, a user must be able to check the results and identify the correct categories from an extended set of categories, and a user can input characters by hand. Both versions use the same recognition method, but they are trained using different sets of training patterns. Recursive X–Y cut and Voronoi diagrams are used for segmentation; k–d tree and generalized learning vector quantization are used for coarse classification; and the modified quadratic discriminant function is used for fine classification. The system provides an interface through which a user can check the results, change binarization methods, rectify segmentation, and input correct character categories by hand. Evaluation done using a limited number of Nom historical documents after providing ground truths for them showed that the two stages of recognition along with user checking and correction improved the recognition results significantly.     

Bright-field scanning confocal electron microscopy using a double aberration-corrected transmission electron microscope

Scanning confocal electron microscopy (SCEM) offers a mechanism for three-dimensional imaging of materials, which makes use of the reduced depth of field in an aberration-corrected transmission electron microscope. The simplest configuration of SCEM is the bright-field mode. In this paper we present experimental data and simulations showing the form of bright-field SCEM images. We show that the depth dependence of the three-dimensional image can be explained in terms of two-dimensional images formed in the detector plane. For a crystalline sample, this so-called probe image is shown to be similar to a conventional diffraction pattern. Experimental results and simulations show how the diffracted probes in this image are elongated in thicker crystals and the use of this elongation to estimate sample thickness is explored.     

TMN implementation strategy based on OSI management standards

In order to facilitate the implementations ofTMN interface protocols/services studied inITU-T, it is very important to define profiles for supportingTMN management service. This paper proposes a concrete method for achieving this based on osi management standards as a promisingTMN implementation method. It proposes an idea of structuring theTMN ISP’S based on the structure of the osi managementISP’S. The paper discusses aTMN based on the osi managementISP’S. Finally the implementation as software is discussed and a software architecture for efficient application development is proposed.