6-Phospho-{beta}-galactosidases of Gram-positive and 6-phospho-{beta}-glucosidase B of Gram-negative bacteria: comparison of structure and function by kinetic and immunological methods and mutagenesis of the lacG gene of staphylococcus aureus

The 6-phospho-ß-galactosidase of Staphylococcus aureus,Lactococcus lactis and Lactobacillus casei and 6-phospho-ßglucosidaseB of Escherichia coli build a subfamily inside a greater enzymefamily, named the glycosal hydrolase family 1, which, hi addition,contains nine ß-glycosidases of different origins.Kinetic and immunological evidence is provided in this reportwhich strengthens the relationship of the four 6-phospho-ß-glycosidases.It is shown that the 6-phospho-ß-galactosidases and6-phospho-ß-glucosidase B are able to split aromaticß-galactoside phosphates and ß-glucosidephosphates. The turnover numbers of hydrolysis of substrateswith different epimerization at C-4 of the glycon vary up to15-fold only. Two polydonal antisera, one derived against thenative 6-phospho-ß-galactosidase from S.aureus andthe other derived against the 6-phospho-ß-glucosidaseB, cross-reacted with both enzymes. Peptides of the proteinswere separated by reverse phase HPLC. The cross-reacting peptideswere sequenced and shown to be localized at almost the sameposition in the aligned primary structures of both enzymes.An insertion of nine amino adds near these antigenic domainsis unique for the 6-phospho-ß-glycosidases and missingwithin the sequences of the ß-glycoside-specific membersof the family. The lacG gene of a 6-phospho-ß-galactosidasenegative S.aureus mutant was doned into E.coli and sequenced.In the totally inactive mutant protein only the glycine at position332 was changed to an arginine. This amino acid is part of thesequence insertion near the antigenic domain reacting with bothantisera. These data support the assumption that the regionis of great importance for the function of the enzymes and thatit is possible it determines the specificity of the phosphorylatedform of the substrates. In addition, the 6-phospho-ß-galactosidaseof S.aureus was modified by sitedirected mutagenesis of thecorresponding lacG gene hi order to replace residues Glul60and Glu375, which were suspected of being involved hi the generalacid catalysis of substrate hydrolysis, with glutamine residues.The mutant protein 160EQ retained some catalytic activity whilethe protein 375EQ was totally inactive. Glu375 is the activesite nudeophile of the 6-phospho-ß-galactosidase ofS.aureus. It is located in the sequence motif ENG where Glu358was identified as the catalytkally active nudeophile hi theß-glucosidase of Agrobacterium.     

Comprehensive meta-analysis of integrity test validities: Findings and implications for personnel selection and theories of job performance.

Conducted a comprehensive meta-analysis based on 665 validity coefficients across 576,460 data points to investigate whether integrity test validities are generalizable and to estimate differences in validity due to potential moderating influences. Results indicate that integrity test validities are substantial for predicting job performance and counterproductive behaviors on the job, such as theft, disciplinary problems, and absenteeism. The estimated mean operational predictive validity of integrity test for predicting supervisory ratings of job performance is .41. Results from predictive validity studies conducted on applicants and using external criterion measures (i.e., excluding self-reports) indicate that integrity tests predict the broad criterion of organizationally disruptive behaviors better than they predict employee theft alone. Despite the influence of moderators, integrity test validities are positive across situations and settings. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

From dust till drawn

We present a system for drawing pastel media in real-time as an effective alternative to most existing digital solutions that basically allow for drawing arbitrary strokes in a particular style. Our approach is focused on the simulation of the natural material itself and on its interaction with the drawing surface and the drawing tool. Upon free-form drawing, a bidirectional transfer of pigment takes place. In one direction, the paper surface is dusted with new pigment particles broken off the tip (i.e., the end of the drawing tool). A large part of these particles will be deposited or blended together with previously deposited ones whereas the remainder does not contribute to the drawing and is blown off. On the other hand, a certain amount of previously deposited pigment is scraped off and picked up again soiling the tip. This is noticeable in the next strokes to be drawn. Furthermore, both the tip and the paper surface are subject to weathering depending on the exerted pressure and friction of the drawing tool, and the bumpiness of the paper. As a result, the paper surface becomes slightly damaged, limiting the deposition of new pigment. The tip, on the other hand, becomes blunt making new strokes wider. From a stylistic point of view, similarly to traditional drawings our results convey the artists’ characteristics (e.g., the way of wielding the brush, skillfulness, feeling for the medium). Therefore, we believe that our system allows an artist to create realistically looking pastel images without losing his/her personal touch.     

NEDAP-Wahlcomputer – Manipulationsmethoden an Hard- und Software

In Deutschland geh?rt der Einsatz von NEDAP-Wahlcomputern in immer mehr Gemeinden zum Alltag. Durch Untersuchungen zur Sicherheit und Manipulierbarkeit konnten Erkenntnisse gewonnen werden, mit welchen Methoden manipulierte Software in den Wahlcomputer eingebracht und Hardwarekomponenten ausgetauscht werden k?nnen. Dadurch ist erstmals eine Beurteilung der Eignung der NEDAP-Wahlcomputer hinsichtlich der Anforderungen an manipulationsfeste, nachvollziehbare und transparente Wahlen m?glich.     

CMOS scaling into the nanometer regime

Starting with a brief review on 0.1-μm (100 nm) CMOS status, this paper addresses the key challenges in further scaling of CMOS technology into the nanometer (sub-100 nm) regime in light of fundamental physical effects and practical considerations. Among the issues discussed are: lithography, power supply and threshold voltage, short-channel effect, gate oxide, high-field effects, dopant number fluctuations and interconnect delays. The last part of the paper discusses several alternative or unconventional device structures, including silicon-on-insulator (SOI), SiGe MOSFET's, low-temperature CMOS, and double-gate MOSFET's, which may lead to the outermost limits of silicon scaling     

A fast analog mean-timer

A circuit is described that determines the average of the transit time of light from an event in a scintillation counter to the two ends of that counter. The design offers superior time resolution and a novel gating scheme that significantly reduces circuit dead-time.     

Energy-efficient penetration and perforation of targets in the hypervelocity regime

In general, the performance of a KE penetrator against most targets increases with velocity regardless of the particular penetrator-target interaction mode. It is possible to show that there exists an optimum velocity which maximizes the performance of an impacting penetrator for a given expenditure of kinetic energy. Simple graphical methods are described that determine the optimum velocity from general performance-velocity plots. These graphical methods may also be applied to experimental data alone. In addition, simple analytic models which describe the velocity dependent penetration/perforation performance of KE penetrators are examined and extended. These models may be used to explicitly assess the influence of parameters such as target strength and density and penetrator mass, strength and density. For some of the models in this analysis, the explicit relations between the optimum striking velocity for a specific kinetic energy value and the penetrator-target parameters are described.     

Trade secrets in software design and logic

Much of what has been written in Australia about the protection of computer programs has been inspired by high profile cases concerning the boundaries of copyright protection. The Australian law firm Clayton Utz says that there has been little discussion about other types of legal protection for computer programs, and as a consequence, other intellectual property laws should not be overlooked.     

A review of contractor selection practice in the U.K. construction industry

Background, prime influences upon present day methods and current trends in United Kingdom tendering practices are reviewed. Significant weaknesses are identified, confirming the need for some revision of existing contractor selection methods. The findings compliment recommendations of the recent government/industry review of the construction sector headed by Sir Michael Latham. Specifically, that tendering procedure should embrace investigation of contractors' potential to deliver a service of acceptable standard, on time, within budget. Present selection methods often fail in this objective, being inclined to discriminate predominantly on cost.