Insertional re-activation of a chloramphenicol acetyltransferase misfolding mutant protein

The deletion of nine residues from the C-terminus of the bacterialchloramphenicol acetyltransferase (CAT) results in depositionof the mutant protein in cytoplasmic inclusion bodies and lossof chloramphenicol resistance in Escherichia coli. This foldingdefect is relieved by C-terminal fusion of the polypeptide withas few as two residues. Based on these observations, efficientpositive selection for the cloning of DNA fragments has beendemonstrated. The cloning vector encodes a C-terminally truncatedCAT protein. Restriction sites in front of the stop codon allowthe insertion of target DNA, resulting in the production ofproperly folded CAT fusion proteins and regained chloramphenicolresistance. The positive selection of recombinants is accomplishedby growth of transformants on chloramphenicol-containing agarplates. The method appears particularly convenient for the cloningof DNA fragments amplified by the PCR because minimal informationto restore CAT folding can be included in the primers. The cloningof random sequences shows that the folding defect can be relievedby fusion to a wide variety of peptides, providing great flexibilityto the positive selection system. This vector may also contributeto the determination of the role of the C-terminus in CAT folding.     

Active,passive and transpassive dissolution of a nickel base super alloy in concentrated acid mixture solution

The electrochemical and corrosion behaviour of a nickel base super alloy (C-263) has been investigated in the deaerated binary and ternary solution mixture of concentrated phosphoric acid, acetic acid, sulphuric acid, nitric acid or water using potentiostatic technique at 35°C. The possibilities of electropolishing of this alloy in these solution mixtures have been also explored. The alloy showed distinct active, passive and transpassive behaviour in the experimental solutions. The alloy remained active and turned passive in the negative potential region. Transpassive dissolution of the alloy is observed and electropolishing is achieved in this region. The best electropolishing is obtained in 50% H₃PO₄ + 40% CH₃COOH + 10% H₂SO₄. Higher content of water in the electrolytic solution is not useful for electropolishing of the alloy The experimental results also suggest that a current plateau in the transpassive potential region is not a sufficient condition to achieve electropolishing.     

Characterization of allergoids from ovalbumin in vitro and in vivo

Several in vivo and in vitro methods for monitoring immunological properties of two allergoids obtained by formaldehyde treatment of ovalbumin (OA) were developed. The calculated molecular weight of allergoids was 80 kD (OA-F1) and 165 kD (OA-F2), respectively. The allergenic activity in vitro of allergoids in mast-cell histamine release assay was 1000 times lower than of OA. Both allergoids showed reduced ability to induce passive cutaneous anaphylaxis in the Sprague-Dawley rats or systemic anaphylaxis in Dunkin-Harley guinea-pigs. The ability of OA and allergoids to bind to the OA-specific IgE antibodies was measured in vivo by the inhibition of passive cutaneous anaphylaxis (PCA-inhibition). Allergoid binding to IgE was 51-66% lower than the native allergen. Moreover, the avidity of OA-specific IgG antibodies, measured by ELISA-inhibition, for allergoids and allergen was of the same order. Allergoids induced a different pattern of humoral immune response from that, induced by the native allergen. Thus, after immunization of BALB/c mouse, both allergoids induced a higher production of IgG and a lower production of IgE than OA, only OA-F2 induced a lower production of IgG1. The differences in the IgA response to the immunogens was not significant. Delayed hypersensitivity studies in the BALB/c mouse showed that allergoids were 5- to 12-times less effective in inducing a cell-mediated immune response than OA. The present study provides a battery of immunological methods for preclinical testing of modified allergens.     

Afterburner [network-independent card for protocols]

Many current implementations of protocols such as the Transmission Control Protocol/Internet Protocol (TCP/IP) are inefficient because data are often accessed more frequently than necessary. Three techniques that reduce the need for memory bandwidth are proposed. The techniques are copy-on-write, page remapping, and single-copy. Afterburner, a network-independent card that provides the services that are necessary for a single-copy protocol stack, is described. The card has 1 MByte of local buffers and provides a simple interface to a variety of network link adapters, including HIPPI and asynchronous transfer mode (ATM). Afterburner can support transfers to and from the link adapter card at rates up to 1 Gbit/s. An implementation of TCP/IP that uses the features provided by Afterburner to reduce the movement of data to a single copy is discussed. Measurements of the end-to-end performance of Afterburner and the single-copy implementation of TCP/IP are presented     

Functional Properties and Food Applications of Rapeseed Protein Concentrate

Rapeseed protein concentrate (RC), prepared with 2% hexameta-phosphate, was tested for its functionality and performance in some foods. The RC had good nitrogen solubility, fat absorption, emulsification, and whipping capacities but poor water absorption and gelling properties. It increased the emulsion stability, and protein but lowered the fat content of wieners. It also increased the cooking yield, reduced the shrinkage and tenderized meat patties. Results were similar to soybean isolate except for the poorer color and flavor. The cooking yield of RC supplemented wieners was less than the all-meat control and soybean-supplemented wieners. A 9% RC dispersion mixed with an equal volume of eggwhite produced a meringue of comparable stability and texture to that of eggwhite alone.     

Interdisciplinary research: roles for self-organization

No generally accepted principles and guidelines currently exist to help engineers design local interaction mechanisms that result in a desired global behavior. However, several communities have developed ways of approaching this problem in the context of niched application areas. Because the ideas underlying these approaches are often obscured or underemphasized in technical papers, the authors review the role of self-organization in their work. They provide a better picture of the status of the emerging field of self-organizing systems or autonomic computing.     

A systematic techno-economic assessment of a WEC system installation for low-cost electrification of a habitable dwelling

This paper deals with the technical feasibility and economic viability of a grid connected wind-energy conversion system (WECS) used to cover the energy demand of an average habitable dwelling. The various parts of the wind-energy system are described and the useful electrical energy production is determined using a simulation program based on the Monte Carlo method. An economic analysis of the WEC system is performed using a computerized assessment tool. Important financial indices are calculated and financial scenarios investigated.     

Advanced Method for Single Crystal Casting of Turbine Blades for Gas Turbine Engines and Plants

A method for fabricating single crystal blades that combines the techniques of seed crystals and selection is suggested. The method realizes the advantages of both techniques, i.e., the high structural perfection and the possibility of fabricating single crystals with specified spatial orientation. Metallographic and x-ray diffraction analyses are used to study the processes of nucleation of the single crystal structure of blade castings fabricated from high-temperature nickel alloys by the method of selection and seed crystals. A commercial process for fabricating cast single crystal turbine blades by the new method is suggested.