Analysis of enterococci using portable testing equipment for developing countries--variance of Azide NutriDisk medium under variable time and temperature.

This report compares the enterococci count on samples obtained with Azide NutriDisk (AND) (sterile, dehydrated culture medium) and Slanetz and Bartley (SB) medium when exposed to a variable in incubation time and temperature. Three experiments were performed to examine the recovery of enterococci on AND and SB media using membrane filtration with respect to: (a) incubation time; (b) incubation temperature; and (c) a combination of the two. Presumptive counts were observed at 37, 41, 46 and 47 degrees C and at 20, 24, 28 and 48 h. These were compared to AWWA standard method 9230 C (44 degrees C, 44 h). Samples were confirmed using Kanamycin Aesculin Azide (KAA) agar. Friedman's ANOVA and Students t-test analysis indicated higher enumeration of enterococci when grown on AND (p = 0.45) than SB (p = < 0.001) at all temperatures with a survival threshold at 47 degrees C. Significant results for AND medium were noted at 20 h (p = 0.021), 24 h (p = 0.278) and 28 h (p = 0.543). The study concluded that the accuracy of the AND medium at a greater time and temperature range provided flexibility in incubator technology making it an appropriate alternative to SB medium for monitoring drinking water using field testing kits in developing countries.     

EBSP study of the annealing behavior of aluminum deformed by equal channel angular processing

Commercial purity aluminum (99.5%) was fabricated by equal channel angular pressing (ECAP) up to total accumulated strains of approx. 10. The annealing behavior of material deformed to total strains of approx. 1 and 10 was investigated, using heat treatments of 2 h at various temperatures from 100 to 500 °C. The microstructure of the annealed materials was characterized using the electron back-scatter pattern technique. A number of parameters were determined including the distribution and average values of both the boundary spacings and misorientations. For samples deformed to a total strain of 1, annealing resulted in discontinuous recrystallization. For samples deformed to a total strain of 10, annealing resulted in microstructures exhibiting characteristics of both uniform coarsening and, in a number of places, of discontinuous recrystallization. An attempt was made, based on the boundary spacing distributions, to separate these two components. The grain size after annealing was still however small, being just 6.4 μm after 2 h at 300 °C.     

Modeling tool wear in end-milling using enhanced GMDH learning networks

This paper presents an enhanced approach to predictive modeling for determining tool-wear in end-milling operations based on enhanced-group method of data handling (e-GMDH). Using milling input parameters (speed, feed, and depth-of-cut) and response (tool wear), the data for the model is partitioned into training and testing datasets, and the training dataset is used to realize a predictive model that is a function of the input parameters and the coefficients determined. In our approach, we first present a methodology for modeling, and then develop predictive model(s) of the problem being solved in the form of second-order equations based on the input data and coefficients realized. This approach leads to some generalization because it becomes possible to predict not only the test data obtained during experimentation, but other test data outside the experimental results can also be used. Moreover, this approach makes it easy to present the realized solution in a form that can be further optimized for the input parameters using some optimization techniques. The results realized using our e-GMDH method are promising, and the comparative study presented shows that the e-GMDH outperforms polynomial neural network (PNN); moreover, it is more flexible than the conventional GMDH, which tends to produce nonlinear solutions even for simple problems. In the investigation, the extended particle swarm optimization (PSO) technique was applied to obtain the optimal parameters. Consequently, the modeling approach is extremely useful in realizing a computer-aided process-planning system in an advanced manufacturing environment.     

The sleeping dragon: Wind energy in the Asia-Pacific region

The Asia-Pacific region is one of the fastest growing regional economies in the world today. The region is home to over half of the global population and is one of the most important markets for multinational corporations to penetrate. The wind power sector is no exception. Wind resources remain untapped and the demand for electricity continues to grow as a result of expanding and modernizing economies. Godfrey Chua, Emerging Energy Research USA reports on the current status and development of the wind industry in the region.     

Surface localization of the sea urchin egg receptor for sperm

The sea urchin egg receptor for sperm is thought to be involved in species-specific sperm-egg interactions at the egg surface. Recent revisions in the deduced amino acid sequence of the cloned cDNAs indicate that the protein encoded does not possess the common structural hallmarks of a membrane protein. Thus, investigation of the localization and association of the protein with the egg surface is crucial. We describe and characterize a new monoclonal antibody raised against recombinant sperm receptor protein. This antibody, in conjunction with several polyclonal antibodies, was used to study the receptor protein in eggs. Immunoprecipitation studies indicated that the antibodies recognize the high Mr (ca. 350 K) sperm receptor protein which copurified with egg plasma membrane-vitelline layer complexes. The sperm receptor protein was solubilized only by detergents and not by treatments designed to solubilize peripherally associated or lipid-anchored membrane proteins, suggesting a tight association with the membrane fraction. Confocal immunofluorescence microscopy of live eggs indicated surface staining. Finally, lysylendoproteinase C treatment of live eggs resulted in a loss of the high Mr receptor protein epitopes, and the concomitant release of a 70-kDa proteolytic fragment, which correlated with a reduced ability of the eggs to be fertilized. Taken together, these data indicate that at least some fraction of the sperm receptor protein is present on the egg surface, a requisite locale for a sperm binding protein.     

Cytoplasmic tail deletion of T cell receptor (TCR) beta-chain results in its surface expression as glycosylphosphatidylinositol-anchored polypeptide on mature T cells in the absence of TCR-alpha

Surface expression of the T cell antigen receptor (TCR) in mature T cells requires the association of a variable heterodimer (alpha.beta or gamma.delta) with six invariant CD3 polypeptides (gamma, delta, epsilon-epsilon, zeta-zeta, or zeta-eta). We described here that deletion of the cytoplasmic tail polypeptide sequence (Lys-Lys-Lys-Asn-Ser) of TCR beta-chain (beta CT) results in expression of the truncated beta-chain on the surface of a mature T cell hybridoma line, in the absence of TCR-alpha, as a glycophosphatidylinositol (GPI)-anchored monomeric polypeptide. The GPI-anchored TCR-beta CT is not associated with CD3-epsilon and is incapable of conventional signal transduction. Association with TCR-alpha prevents beta CT from GPI-linkage formation. The alpha beta CT heterodimer binds the CD3 polypeptides, and the resultant TCR alpha beta CT/CD3 complex is capable of signal transduction. Our data show that a signal sequence for GPI-linkage formation is present in TCR-beta, and this alternative membrane anchoring mechanism can be utilized by beta-chain polypeptide lacking the CT sequence. We conclude therefore that in the absence of TCR-alpha expression, the beta-chain CT sequence plays an essential function in hindering GPI-linkage formation, thereby preventing escape of incompletely assembled TCR beta-chain to the cell surface of mature T cells.     

The stationary-phase-exit defect of cydC (surB) mutants is due to the lack of a functional terminal cytochrome oxidase

The surB gene was identified as a gene product required for Escherichia coli cells to exit stationary phase at 37 degrees C under aerobic conditions. surB was shown to be the same as cydC, whose product is required for the proper assembly and activity of cytochrome d oxidase. Cytochrome d oxidase, encoded by the cydAB operon, is one of two alternate terminal cytochrome oxidases that function during aerobic electron transport in E. coli. Mutations inactivating the cydAB operon also cause a temperature-sensitive defect in exiting stationary phase, but the phenotype is not as severe as it is for surB mutants. In this study, we examined the phenotypes of surB1 delta(cydAB) double mutants and the ability of overexpression of cytochrome o oxidase to suppress the temperature-sensitive stationary-phase-exit defect of surB1 and delta(cydAB) mutants and analyzed spontaneous suppressors of surB1. Our results indicate that the severe temperature-sensitive defect in exiting stationary phase of surB1 mutants is due both to the absence of terminal cytochrome oxidase activity and to the presence of a defective cytochrome d oxidase. Membrane vesicles prepared from wild-type, surB1, and delta(cydAB) strains produced superoxide radicals at the same rate in vitro. Therefore, the aerobic growth defects of the surB1 and delta(cydAB) strains are not due to enhanced superoxide production resulting from the block in aerobic electron transport.