Vital-Fluorochromization of Microorganisms Using 3′,6′-Diacetylfluorescein to Determine Damages of Cell Membranes and Loss of Metabolic Activity by Ozonation

Cell suspensions of Escherichia coli as a model for bacterial populations in wastewaters were treated with ozone as a disinfectant in a continuously operated pilot plant with a plug flow reactor. Suspensions with an initial number of CFU (colony forming units) of 10⁸ mL^?1 were ozonized with ozone concentrations up to 12 mg/L. Metabolic activities and membrane functions break down with increasing ozone concentrations. The fluorochromization using 3′,6′-diacetylfluorescein (FDA) proved to be a suitable method for the detection of an alteration in permeability of the cell membranes and an inactivation of metabolic activity. By fluorescence microscopic and photometric investigations it could be clearly demonstrated that, in the case of disinfection with ozone, inactivation of the cells includes first of all a damage of the cell membranes. In contrast to the determination of the number of CFU, fluorochromization allows the detection of alteration in metabolic activities.     

Diffusion bonding of an aluminium-lithium alloy (AA8090) using aluminium-copper alloy interlayers

Diffusion bonds have been produced between sheets of an Al-Li-Cu-Mg-Zr alloy using aluminium-4% copper vapour deposited metallic interlayers. Microstructural changes occurred both in the parent alloy and in the bond interface after diffusion bonding cycles and post-bonding heat treatments were analysed. Different metallographic techniques (light microscopy, scanning and transmission electron microscopy) have been used. Diffusion bonding trials were carried out using the same alloy (AA8090), both in non-superplastic (T6) and superplastic conditions. Differences in their behaviours in relation to diffusion bonding were observed.     

Air bearing design, optimization, stability analysis andverification for sub-25 nm flying

Three sub-25 nm fly height sliders are presented for near contact recording. The designs are geared towards the goal of achieving 10 Gb/in ² areal density. The optimization procedure presented shows promise for facilitating achievement of this goal. The dynamic simulations show the stability of these designs when disturbed from their steady state conditions     

Magnetometric determination of the angular position of an object

A method is described for finding the angular position of an object by measuring at two selected points in space the components of an alternating magnetic field set up by two sources. The angular position of the object and its coordinates can be measured even if the object is moved or rotated about any axis through 0 to 360° in the presence of magnetic interference. Translated from Izmeritel'naya Tekhnika, No. 12, pp. 34–37, December, 1996.     

Neural networks for contract bridge bidding

The objective of this study is to explore the possibility of capturing the reasoning process used in bidding a hand in a bridge game by an artificial neural network. We show that a multilayer feedforward neural network can be trained to learn to make an opening bid with a new hand. The game of bridge, like many other games used in artificial intelligence, can easily be represented in a machine. But, unlike most games used in artificial intelligence, bridge uses subtle reasoning over and above the agreed conventional system, to make a bid from the pattern of a given hand. Although it is difficult for a player to spell out the precise reasoning process he uses, we find that a neural network can indeed capture it. We demonstrate the results for the case of one-level opening bids, and discuss the need for a hierarchical architecture to deal with bids at all levels.     

A preliminary investigation into the microscopic depassivation of passive titanium implant materials in vitro

A sensitive electrochemical technique has been used to examine the passive state of titanium-based materials in Ringer's physiological solution. At ambient temperature, the alloy Ti-6Al-4V shows transient microscopic breakdown of the passive state induced by the presence of chloride ions, and enhanced by increased acidity. These breakdown events involve highly localized depassivation of the passive surface followed by repassivation. Under similar experimental conditions no breakdown of passive titanium was detected.     

Reconstitution of CD3 zeta coupling to calcium mobilization via genetic complementation

The integrity of the T cell receptor complex (CD3-TCR) transduction machinery is central to T cell development and to T cell effector function. Molecular dissection of the multimeric CD3-TCR complex revealed that at least two associated polypeptides, CD3 zeta and CD3 epsilon, autonomously couple antigenic recognition event to early and late events of the intracytoplasmic activation cascade. A 18-amino acid motif based on a tandem YXXL stretch, the activation receptor homology sequence 1 (ARH-1) motif, is necessary and sufficient to the transducing properties of both CD3 zeta and CD3 epsilon. Stimulation of chimeric molecules made of ecto- and transmembrane domains of various cell surface proteins and intracytoplasmic domains of CD3 epsilon or CD3 zeta leads to an increase in the intracellular Ca2+ concentration ([Ca2+]i) in Jurkat cells. We describe here that a similar CD25/zeta chimeric molecule was unable to induce a detectable [Ca2+]i rise upon CD25 cross-linking once expressed in the murine thymoma BW-. A Ca2+ influx could, however, be triggered in BW- cells by thapsigargin, i.e. following depletion of Ca2+ stores. Somatic cell hybrids made from BW- and either thymocytes or mature lymph node T cells reconstituted the coupling of CD3 zeta to the Ca2+ signal via an ARH-1 motif-dependent pathway. However, pervanadate-induced Ca2+ mobilization, a phenomenon attributed to tyrosine phosphorylation, was impaired in BW-cells and reconstituted in hybridomas. In contrast to the Ca2+ response, IL-2 production was induced in both BW- and hybrids cells, which questions the functional relevance of [Ca2+]i augmentation in T cell activation. In conclusion, the properties of the BW- thymoma, which define a novel group of CD3 zeta transduction cell mutants, as well as its complementation by somatic cell fusion demonstrate that this cell line represents a useful model to dissect the signaling pathway that couples CD3 zeta to Ca2+ mobilization by genetic reconstitution.