The band areas of proteins determined by fluorescent scanning in the commercial automated gel electrophoresis apparatus

An automated gel electrophoresis apparatus, recently available commercially, allows one to follow the band during electrophoresis in real time, and lends itself therefore to an evaluation of bandwidth as a function of migration time (the dispersion coefficient), resolution and band shape. These determinations assume the constancy of band area with migration time and at various gel concentrations. The purpose of the present study was to verify these assumptions. Representative proteins and sodium dodecyl sulfate (SDS)-proteins, either natively fluorescent or fluorescein carboxylate labeled, were found to exhibit band areas which approach constancy as a function of migration time in both agarose and polyacrylamide gel electrophoresis, provided that (i) the protein concentration under the band was low enough to obviate self-quenching of fluorescence; (ii) the separation of the protein of interest from contaminants had progressed sufficiently during the time at which band areas were measured; (iii) the baseline under the peak was sufficiently well defined. However, band areas decrease with increasing gel concentration. Protein peaks exhibited leading and trailing tails. The ratio of the combined tail area to total area appeared to be near-constant at varying migration times. However, that ratio increases with increasing gel concentration. The tail area does not appear to be an artifact of fluorometric detection since it is reproduced upon fluorimetric analysis of the protein eluted from gel slices after electrophoresis. However, it may be due to photochemical destruction under the conditions of repetitive fluorometric peak detection.     

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.     

A 3-D motion tracing system in High School Physics teaching

Abstract This article presents a study of a large scale incorporation of one data acquisitionsystem into physics teaching in Israel. By 1994 about 30% of the schools throughout the country used the V-scope, a 3-D multibody motion tracing system, in their lab programme. The views and reactions of post-training teachers, experienced teachers and students on various aspects of using the system are described, analysed and compared.     

Static-state feedback control of discrete-event systems underpartial observation

Studies static-state feedback control of discrete-event systems under partial observation. The authors present a necessary and sufficient condition for the existence of a static-state feedback controller. A static controller is a special case of a dynamic controller studied by Kumar et al. The authors' condition, however, has computational advantage in contrast to those obtained by Kumar et al. The authors also present a necessary and sufficient condition under which the modular feedback synthesis is possible     

Providing information to clients about psychological tests: A survey of attorneys' and law students' attitudes.

Surveyed 70 practicing attorneys and 150 law students to determine the extent to which they felt it was their responsibility to inform clients they referred for psychological testing about the testing process and about the validity scales on tests. Results showed that almost 50% of the attorneys and over 33% of the students believe that clients referred for testing always or usually should be informed of validity scales on tests. These results suggest that many test takers, particularly in forensic settings, may have information about validity scales that facilitates their giving biased responses without detection. Psychologists doing forensic evaluations should be alert to this possibility and should consider using more than one approach to detect response distortions. (PsycINFO Database Record (c) 2010 APA, all rights reserved)