A stereoselective cobalt-containing nitrile hydratase

Nitrile hydratase from Pseudomonas putida NRRL-18668 has been purified and characterized. The purified enzyme catalyzes the hydration of 2(S)-(4'-chlorophenyl)-3-methylbutyronitrile at least fifty times faster than that of 2(R)-(4'-chlorophenyl)-3-methylbutyronitrile. This enzyme is a member of the class of nitrile hydratase that contains cobalt. Visible absorption and CD spectra suggest the cobalt exists as a non-corrin low-spin Co3+ ion in a tetragonally-distorted octahedral ligand field. Chemical reduction of the native enzyme results in a species with the EPR signature of a low-spin Co2+ complex. Like the other cobalt-containing nitrile hydratases, this enzyme is relatively stable, maintaining its activity below 35 degrees C, and it shows a broad activity optimum between pH 7.2 and 7.8. The structural genes for this enzyme have been cloned and sequenced. The deduced amino acid sequences for the alpha and beta subunits show 48-63% and 35-41% homology, respectively, to other sequenced nitrile hydratases. In particular, the cysteine residues in the alpha subunit that have been suggested to coordinate the metal ion in the iron-containing nitrile hydratases [Brennan, B. A., Cummings, J. G., Chase, D. B., Turner, I. M., Jr., & Nelson, M. J. (1996) Biochemistry 35, 10068-10077] are conserved in this enzyme, suggesting that this nitrile hydratase, like the enzyme from Rhodococcus rhodochrous J1, is a member of a newly described class of metalloenzymes with Co3+-thiolate ligation [Brennan, B. A., Alms, G., Nelson, M. J., Durney, L. T., & Scarrow, R. C. (1996) J. Am. Chem. Soc. 118, 9194-9195].     

Validation of analytical capillary electrophoresis methods for use in a regulated environment

Since its introduction into the analytical laboratory, CE has had to prove that it was capable of generating results comparable to HPLC or GC techniques in the six areas (specificity, precision, accuracy, linearity, ruggedness, and range) typically required of validated methods intended for submission to governmental agencies. This paper will showcase the development and validation of two analytical methods: one for specific identification of HEPES (N-[2-hydroxyethyl]piperazine-N'-[2-ethane sulfonic acid]) and the other to quantitate millimolar concentrations of tris (tris[hydroxymethyl]aminomethane) in high electrolyte solutions. Utilizing neutral markers and internal standards, results for HEPES demonstrate that migration time reproducibility, expressed as %RSD of 2% or less on a variety of capillaries, is obtainable. Additionally, for quantitation of tris, the values obtained for accuracy (%relative mean bias), precision (%RSD), and linearity (r2) over multiple days and capillaries meet the rigorous standards we require of HPLC or GC methods.     

Transoesophageal echocardiographic assessment of haemodynamic function during laparoscopic cholecystectomy

We have measured cardiovascular changes associated with insufflation of carbon dioxide and the reverse Trendelenburg position during laparoscopic cholecystectomy, using transoesophageal echocardiography in 13 healthy patients. End-tidal carbon dioxide values increased after insufflation of carbon dioxide, with values significantly (P < 0.05) increased after lateral tilt positioning. Creation of a pneumoperitoneum was associated with increases (P < 0.05) in left ventricular end-systolic wall stress, concomitant with increases (P < 0.01) in peak airway pressure and systemic arterial pressure. In addition, left ventricular end-diastolic area decreased (P < 0.05) after reverse Trendelenburg positioning. Left ventricular ejection fraction was maintained throughout the study.     

Achieving synergy in collaborative education

Professional software engineering continues to face problems that involve large-scale (complex) software systems development. Such efforts typically require team collaboration over months, possibly years. Once implemented, the system must be maintained for sometimes decades. This development and maintenance life cycle requires not only technical knowledge and skills in software development, but also the knowledge and skills associated with software process. We believe the traditional computer science curriculum fails to consider this spectrum of needs. We also believe, however, that software engineering must use certain elements of the core knowledge and skills taught by computer scientists as its foundation. We describe lessons learned from the 10-year collaboration between Monmouth University and the Software Engineering Center of the US Army's CECOM (Communications Electronics Command). The center recruits electrical engineering and computer science graduates from academic institutions in the geographic regions surrounding its various installations to be part of its intern program; Monmouth University, in turn, incorporates the demands of the intern program in its Master of Science in software engineering curriculum     

Screen‐printed biosensor for allergens

Allergen levels in indoor environments, leading to many diseases, eg asthma, rhinitis and conjunctivitis, affect a large and increasing fraction of the population. A quite effective and inexpensive method of a rough but very rapid overall assessment of total allergen level in the environment has been developed. The method involved estimation of protein in allergen extracts by screen‐printed electrodes using two different techniques. The biosensor comprised a rhodinised carbon working electrode, a silver/silver chloride reference electrode and a carbon counter electrode. In the first method the enzyme protease reacted with allergen protein to release amino acid, which produced hydrogen peroxide in the presence of amino acid oxidase. This was detected amperometrically. The second method used potassium bromide as electrolyte and the electrode was subjected to dual potential. Bromine, released due to electrolysis at higher potential, was consumed by the allergen protein at lower potential. In the first method, a unique technique was used to microencapsulate the enzyme protease and immobilise it on the surface of the electrode by in‐situ polymerisation to avoid contact with the amino acid oxidase. A total of seven allergens were tested and the results gave a good correlation with the standard protein measurement method. Environmental specimens from indoors, schools and workplaces can be evaluated for the aeroallergens produced by dust mites, animal hairs, cockroach debris, pollens, etc as a means of determining the exposure risk. Copyright © 2005 Society of Chemical Industry     

Integrating heterogeneous distributed COTS discrete-event simulation packages: an emerging standards-based approach

This paper reports on the progress made toward the emergence of standards to support the integration of heterogeneous discrete-event simulations (DESs) created in specialist support tools called commercial-off-the-shelf (COTS) discrete-event simulation packages (CSPs). The general standard for heterogeneous integration in this area has been developed from research in distributed simulation and is the IEEE 1516 standard The High Level Architecture (HLA). However, the specific needs of heterogeneous CSP integration require that the HLA is augmented by additional complementary standards. These are the suite of CSP interoperability (CSPI) standards being developed under the Simulation Interoperability Standards Organization (SISO-http://www.sisostds.org) by the CSPI Product Development Group (CSPI-PDG). The suite consists of several interoperability reference models (IRMs) that outline different integration needs of CSPI, interoperability frameworks (IFs) that define the HLA-based solution to each IRM, appropriate data exchange representations to specify the data exchanged in an IF, and benchmarks termed CSP emulators (CSPEs). This paper contributes to the development of the Type I IF that is intended to represent the HLA-based solution to the problem outlined by the Type I IRM (asynchronous entity passing) by developing the entity transfer specification (ETS) data exchange representation. The use of the ETS in an illustrative case study implemented using a prototype CSPE is shown. This case study also allows us to highlight the importance of event granularity and lookahead in the performance and development of the Type I IF, and to discuss possible methods to automate the capture of appropriate values of lookahead.     

Freestanding Alumina Membrane by Double-Layer Anodization

A novel method has been developed for fabricating full or partial freestanding anodic alumina. In our method a sacrificial metal layer is introduced between an Al film and a Si₃N₄ substrate. A freestanding alumina film at wafer scale is successfully achieved by anodizing the double metal layer, during which the alumina is spontaneously stripped off the Si₃N₄ substrate due to the anodic oxidation of the sacrificial layer. The barrier oxide of the alumina film is effectively removed either by H₃PO₄ dissolution or by CF₄ reactive ion etching. The freestanding alumina film is utilized as a contact mask to transfer its nanoporous pattern to a Si substrate. By patterning the sacrificial metal layer with contact lithography, a partial freestanding alumina film is successfully achieved on the silicon chip, producing a unique micro/nanofluidic channel. Compared with previous techniques, the method reported here is advantageous for its simplicity and flexibility     

A Novel Inverse-Magnetostrictive Force Sensor

The change in magnetic permeability of a material under stress (inverse magnetostriction) offers the potential for a high-performance, low-cost force sensor capable of being used in harsh real-world environments. The existing force sensor technologies are limited in their use in commercial products by either cost issues or susceptibility to electromagnetic noise. Inverse Magnetostriction has been used to measure strain in controlled environments since its discovery by Joule in 1847, but not in practical applications due to a lack of data on how magnetic material properties change with environmental conditions such as temperature. Utilizing an innovative noise-reducing self-inductance design, this paper presents the basis for an inverse-magnetostrictive compressive load sensor. A lumped-parameter model for the change in sensor inductance under load, due to both mechanical and magnetic effects, is derived. The material properties of a magnetostrictive iron alloy are empirically determined over a broad range of loads and temperatures. The model and material properties are confirmed by testing a prototype force sensor. The prototype measures compressive forces from 100 to 25 000 N over a temperature range of 20 degC to 120 deg with a typical error of +/-2% (4% max). The sensor does experience significant thermal hysteresis for which the model does not currently account. This work was motivated by the need for a force sensor in an automobile electric brake system and used a single iron alloy (50% Ni), but the model and testing procedure provide a roadmap for future research to improve the performance and capabilities of such a sensor     

Turning software into a service

The software as a service model composes services dynamically, as needed, by binding several lower-level services-thus overcoming many limitations that constrain traditional software use, deployment, and evolution.     

Creep in polyethylene and hydroxyapatite reinforced polyethylene composites

Isochronous stress-strain relationships and long term creep performance for unfilled and hydroxyapatite filled polyethylene composites have been studied. The tests were carried out in a buffered (pH=7.5) Ringer's solution at 37°C. It was observed that the inclusion of hydroxyapatite does not remove the non-linear viscoelasticity of polyethylene. The creep resistance is found to increase with increase in volume fraction of hydroxyapatite. The creep failure of composites at long times can occur due to debonding of the interface.