Preparation of compound-specific and group-specific antibodies to 7-methylguanine and related 7-alkylguanines and their use in immunoaffinity purification

The preparation and characteristics of compound-specific and group-specific antibodies against 7-alkylguanines (7-alkGua) are described. A compound-specific antibody against 7-methylguanine was prepared using a hapten bound to carrier protein through the N2 position. In a competitive enzyme-linked immunosorbent assay (ELISA) 7-methylguanine (7-MeGua) showed 50% inhibition (I50%) at 10 pmol/well at room temperature, but the inhibition was found to be 40 times better at 4 degrees C (I50% at 250 fmol/well). When the antibody was bound to protein A-Sepharose CL4B 7-MeGua was retained in immunoaffinity columns. A group-specific antibody to 7-alkGua was prepared using 7-(2-carboxyethyl)guanine (7-CEGua) bound to carrier protein via the carboxyl group. In a competitive ELISA, this antibody cross-reacted well with 7-CEGua, 7-ethylguanine (7-EtGua), 7-(2-hydroxyethyl)guanine (7-HOEtGua) and 7-(2',3'-dihydroxy)-propylguanine (7-DHPGua) and some inhibition was seen with 7-MeGua. Immunoaffinity columns prepared from this antibody retained a number of 7-alkGua of diverse structure. 7-EtGua in calf thymus DNA treated with diethylsulphate and ethylnitrosourea was isolated by immunoaffinity purification and quantified by HPLC-fluorescence. These results illustrate the potential of immunoaffinity purification for both individual DNA adducts and groups of adducts.     

Mathematical analysis of viscometric (polymer) flow fields in capillaries: Taylor dispersion revisited

A general formulation for capillary flow of two miscible fluids – one a dilute plug of polymer fluid inserted into a fully developed Poiseuille flow of the other, a Newtonian stream – is examined for its long time behavior. Phenomenologically, the system evolves from an initial state, that of a plug within the boundaries of sharp, well defined fronts inside a Newtonian stream, to a more homogenized state in the very long time scale. This problem was addressed by G.I. Taylor but with regard to a system of two Newtonian fluids, leading to the well-known results commonly described as `Taylor axial dispersion'. In this paper, a general and systematic perturbation analysis is presented from which Taylor's result is recovered as a special case of a more general solution which applies to fluids incorporating elastic properties. In particular, the influence of viscoelasticity and (polymer) diffusivity on the observed pressure profile in the capillary conduit is examined. This effect is clearly separated out for small Peclet number flows using asymptotic and numerical analysis. The results identify the influence of fluid viscosity, elasticity, and diffusivity on the observed pressure profile and form the basis for the improved characterization of polymeric elasticity using capillaries – a finding that is of significant scientific and commercial interest. These results were obtained by the authors as a class of observations resulting from the perturbation analysis of forced-flow capillary devices in viscoelastic fluid property investigation.     

A DUCTILITY EXHAUSTION EVALUATION OF SOME LONG TERM CREEP/FATIGUE TESTS ON AUSTENITIC STEEL

Abstract— A limited number of long term creep/fatigue tests performed on two batches of Type 316 steel and one batch of associated 17Cr8Ni2Mo weld metal are reported. Test durations range from 5000 to 32,000 h and temperatures from 5.50 to 625 ° C. Subsequent metallographic examination shows the failures to be wholly or predominantly intergranular. The results are analysed using a ductility exhaustion approach and it is shown that the endurances obtained are within a factor of two of predicted values. The results confirm that the design approach to creep/fatigue currently being developed in the U.K. and based on ductility exhaustion is likely to be satisfactory.     

Structure of metal site in Cd-substituted His117Gly mutant of azurin with and without addition of imidazole derivatives

The present work uses 111mCd-perturbed angular correlations of gamma-rays (PAC) to investigate the structure of the metal site of the His117Gly mutant of Pseudomonas aeruginosa azurin in aqueous solution and the effect on the structure upon addition of the following exogenous ligands: imidazole, 4-methyl imidazole, 1-methyl imidazole, 2-methyl imidazole and histidine. The nuclear quadrupole interaction of cadmium bound to the mutant without addition of exogenous ligands shows a strong pH dependence with three different nuclear quadrupole interactions consistent with two pKa values at about 7.2 and 8.6 at 2 degrees C. Addition of the imidazole derivatives resulted in a significant change in the PAC spectrum showing that they coordinate. This is in accordance with observations by EPR for the same mutant with copper at the metal site [den Blaauwen, T. & Canters, G. W. (1993) J. Am. Chem. Soc. 115, 1121-1129]. However, whereas EPR and ultraviolet/visual absorption show that the characteristics of the wild-type copper protein are regained by addition of the imidazole derivatives with the exception of the possible bidentates (histidine and histamine), the comparison of the PAC results to model calculations shows that the cadmium ion must be fourfold coordinated in most cases, probably binding an additional water or hydroxide ligand. A fourfold coordination is in contrast to cadmium-substituted wild-type azurin where PAC data inferred a threefold coordination by a Cys and two His residues [Danielsen, E. Bauer, R., Hemmingsen, L., Andersen. M., Bjerrum, M. J., Butz, T., Tr?ger, W., Canters, G. W., Hoitink, C. W. G., Karlsson, G., Hansson, O. & Messerschmidt, A. (1995) J. Biol. Chem. 270, 573-580]     

An improved system for the control of dissolved oxygen in freshwater aquaria

A system for the removal and control of dissolved oxygen (DO) from freshwater was designed and constructed with aquarium-type fish studies in mind. Degassed water was obtained using a partial vacuum of -14 psi, and DO regulated at an aquarium scale using electronically controlled aeration with timed partial water renewal. The degassing system was capable of producing water with approximately 1.7 mg L(-1) DO within 10 min of operation, and 0.55 mg L(-1) after 2h. The control system was capable of maintaining DO levels of ca 0.8 mg L(-1) over 48 h in the absence of aeration and further capable of precisely controlling DO levels as low as 1.16+/-0.002 mg L(-1) (mean+/-SEM) with aeration over a 48 h period.     

Savoring Versus Dampening: Self-Esteem Differences in Regulating Positive Affect.

Five studies examined the hypotheses that when people experience positive affect, those low in self-esteem are especially likely to dampen that affect, whereas those high in self-esteem are especially likely to savor it. Undergraduate participants' memories for a positive event (Study 1) and their reported reactions to a success (Study 2) supported the dampening prediction. Results also suggest that dampening was associated with worse mood the day after a success (Study 2), that positive and negative affect regulation are distinct, that self-esteem is associated with affect regulation even when Neuroticism and Extraversion are controlled (Studies 3 and 4), and that self-esteem may be especially important for certain types of positive events and positive affect regulation (Study 5). (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

A 16-Mb MRAM featuring bootstrapped write drivers

A 16-Mb magnetic random access memory (MRAM) is demonstrated in 0.18-/spl mu/m three-Cu-level CMOS with a three-level MRAM process adder. The chip, the highest density MRAM reported to date, utilizes a 1.42/spl mu/m/sup 2/ 1-transistor 1-magnetic tunnel junction (1T1MTJ) cell, measures 79 mm/sup 2/ and features a /spl times/16 asynchronous SRAM-like interface. The paper describes the cell, architecture, and circuit techniques unique to multi-Mb MRAM design, including a novel bootstrapped write driver circuit. Hardware results are presented.