THE USE OF AMPPD AS AN ALTERNATIVE SUBSTRATE FOR AP-MEDIATED DETECTION OF NONRADIOLABELED DNA PROBES IN EUCALYPTUS-SALIGNA

We present a non-radioactive alternative to Southern's (J. Mol. Biol. 98: 503-517, 1975) DNA-DNA hybridization technique. The use of AMPPD - Disodium 3-(4-Methoxyspiro {1,2-dioxetane-3,2'tricyclo[3.3.1.1(3,7)]decan}-4-yl)phyenyl phosphate as an alternative substrate for AP-mediated detection of digoxigenin-11 dUTP-labeled probes made possible the simple and nonhazardous reuse of blots. We used 0.8 % agarose gels containing 30 mug per lane of Eucalyptus saligna DNA, digested with Eco RI, electrophoresed and blotted on to nylon membranes (Hybond-N, Amersham, UK), using the Southern blotting procedure, and UV irradiated for one minute for DNA fixation. The hybridizations were carried out overnight with digoxigenin labeled random inserts of E. saligna DNA by using the Genius Kit (Boehringer Mannheim). Detection of the DNA-DNA hybrids was performed in the presence of 0.5% blocking agent and the substrates NBT/BCIP were replaced by 0.26 mM AMPPD in the final alkaline assay buffer (50 mul/cm2). After membrane incubation for five minutes at room temperature in a sealed plastic bag, the AMPPD solution was retrieved and stored at 4-degrees-C for reuse. A Kodak X-BRAF QA-S film was pressed firmly onto the bag containing the wet membrane, exposed for two to six hours and then developed. After use, the probes were stripped off and the blots reutilized, three times so far, with the same results.

Plasma immersion ion implantation of nitrogen into H13 steel under moderate temperatures

Ion implantation of nitrogen into samples of tempered and quenched H13 steel was carried out by plasma immersion technique. A glow discharge plasma of nitrogen species was the ion source and the negative high voltage pulser provided 10-12 kV, 60 mu s duration and 1.0-2.0 kHz frequency, flat voltage pulses. The temperatures of the samples remained between 300 and 450 degrees C, sustained solely by the ion bombardment. In some of the discharges, we used a N-2 + H-2 gas mixture with 1:1 ratio. PIII treatments as long as 3, 6, 9 and up to 12 h were carried out to achieve as thickest treated layer as possible, and we were able to reach over 20 mu m treated layers, as a result of ion implantation and thermal (and possibly radiation enhanced) diffusion. The nitrogen depth profiles were obtained by GDOS (Glow Discharge Optical Spectroscopy) and the exact composition profiles by AES (Auger Electron Spectroscopy). The hardness of the treated surface was increased by more than 250%, reaching 18.8 GPa. No white layer was seen in this case. A hardness profile was obtained which corroborated a deep hardened layer, confirming the high efficacy of the moderate temperature PIII treatment of steels. (c) 2005 Elsevier B.V. All rights reserved.

Force-versus-time curves during collisions between two identical steel balls

We describe a numerical procedure for plotting the force-versus-time curves in elastic collisions between identical conducting balls. A system of parametric equations relating the force and the time to a dimensionless parameter is derived from the assumption of a force compatible with Hertz's theory of collision. A simple experimental arrangement consisting of a mechanical system of colliding balls and an electrical circuit containing a crystal oscillator and an electronic counter is used to measure the collision time as a function of the energy of impact. From the data we can determine the relevant parameters. The calculated results agree very well with the expected values and are consistent with the assumption that the collisions are elastic. (C) 2006 American Association of Physics Teachers.

Bactericidal stainless steel

Coatings are largely used in industries. However the development of new materials with improved properties still feeds a continuous need for performance, cost or endurance, the coatings are obtained by a hybrid material, organic-inorganic, and this polymer is applied on metallic, ceramic and glassy surfaces. The material generated in-situ on the desired surface has a nanometric structure. Results in abrasion loss (according ASTM standards) showed that the coatings improve the abrasion resistance of stainless steel by 30%, and also, diminish oxidization and surface rugosity.

Analyses of fatigue data in 4340 high strength steel after cadmium electroplating

Bending fatigue tests were carried out to clarify the effects of heat treatment parameters: temperature and time after cadmium electroplating on a high strength steel, to avoid hydrogen embrittlement. Temperatures heat of 190 degrees C, 230 degrees C, 250 degrees C and 300 degrees C at 3, 8 and 24 hours together with the base material electroplated, with and without heat treatment, resulted in 14 conditions studied with respect to fatigue behaviour. Statistical data analysis was performed to identify the best combination temperature/time regarding fatigue strength of the ABNT 4340 steel and the results obtained revealed that the fatigue strength depend on temperature/time conditions.