Cell cycle-dependent tumor necrosis factor apoptosis

To determine if tumor necrosis factor (TNF)-mediated apoptosis affects cells at defined stages of the cell cycle, WEHI-164/2F (WEHI) cells were synchronized at G0-G1 after 3-day cultures in medium containing RPMI 1640 and 0.5% FCS (RPMI-0.5% FCS). The arrested WEHI cells (60-75% in G0-G1) showed increased sensitivity to TNF killing, measured as 48-h 3-(5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide assays, and 15-h apoptosis by propidium iodide staining and flow cytometry analysis. The TNF killing kinetics of G0-G1-arrested cells was similar to controls, and TNF did not accelerate or retard cell cycle progression of the arrested cells after feeding with fresh RPMI-0.5% FCS. However, TNF inhibited WEHI DNA synthesis as early as 1 h after treatment, and inhibition was proportionate to sensitivity to TNF-induced apoptosis. WEHI cells treated with TNF showed a higher percentage of cells in S phase with concomitant decrease in G0-G1 and G2-M. When cultured for 3-18 h in fresh RPMI-0.5% FCS to allow progression of the G0-G1-arrested cells toward the G1-S boundary, WEHI cells became more sensitive to TNF killing, especially at the 3-9 h time points. Moreover, TNF did not degrade [125I]5-iodo-2'-deoxyuridine-labeled WEHI DNA if the labeled cells were precultured for 9 h in fresh RPMI-0.5% FCS to allow them to pass S phase before the addition of TNF. These results show that TNF-induced apoptosis of WEHI cells is connected to cell cycle events; WEHI targets receive the TNF cytotoxic signal mainly at the G1-S boundary and begin to die by apoptosis as they exit from S phase.     

Dynamic simulations of the molecular conformations of wild type and mutant xanthan polymers suggest that conformational differences may contribute to observed differences in viscosity

Xanthan gum is an exopolysaccharide secreted by the bacterium Xanthamonas campestris whose ability to make solutions viscous at low concentrations and over a pH and temperature range have generated much interest in both academic and industrial environments. Mutant Xanthamonas strains have been derived that produce xanthan gums with an altered or variant subunit chemical structure and different measured viscosities when compared with the wild type (wt) form of the polymer. Two variant gums were targeted as potentially interesting in this study, these being the nonacetylated tetramer (natet) and the acetylated tetramer (atet), which both lack a side-chain terminal mannose residue and in one case (natet) lacks an acetate group on an internal mannose residue. Solutions of these tetrameric gums possess viscosities higher (natet) and lower (atet) than the wt gum, and therefore we have attempted to determine whether these molecules possess unique conformational preferences when compared with the wt and with each other. In this manner we can initiate an understanding of how a polysaccharide's conformation contributes to its solution properties. The GEGOP software permits a sampling of the static and dynamic equilibrium states of carbohydrate molecules, and this software was employed to calculate equilibrium states of representative oligosaccharides with chemical structures representative of xanthan-like molecules. Energy minimization techniques revealed similar local minima for all three molecules. Some of these minima are comprised of elongate backbone conformations (A type) in which side chains fold onto backbone surfaces. Other minima with A backbones possessed side chains in less intimate backbone contact especially when calculations were performed with a low dielectric constant. This phenomenon was particularly pronounced in the wt molecule where an increased number of negatively charged side-chain residues experience charge repulsion resulting in reduced side-chain-backbone contact. Metropolis Monte Carlo (MMC) dynamic simulations performed with an elevated temperature factor (1000 K) allowed a better qualitative representation of conformational space than 300 K simulations. Employing a nonhierarchical cluster analysis method (population density profile: PDP) coupled with a classification scheme, it was possible to partition resulting MMC data sets into conformational families. This analysis revealed that in simulations performed with different dielectric constant values (10, 25, and infinity) all molecules possessed primarily A-type backbones. Less elongate, more open helical backbone forms (B, C, D, J, and Flat-a) did occur during the simulations but were populated to a lesser extent. In the natet molecule significantly open helical backbones existed (E, F, G, H, and I) that did not occur in the lower viscosity wt and atet molecules. PDP clustering methods and subsequent conformational classification applied to the first residue (mannose) of the side chain permitted a determination of side-chain orientation. Comparison of all three molecules indicated a larger population of side-chain conformational families in less direct backbone contact for the wt molecule than either of the variant molecules (natet/atet) suggesting that the side chains in the wt are more flexible. Thus, a major conformational difference between the high viscosity natet and the lower viscosities of the wt/atet is the increased amount of open helical backbone in the natet. In addition, the significant difference between the higher viscosity wt and the lower viscosity atet is the increase side-chain flexibility in the wt. We hypothesize that conformational differences of this kind could form a partial explanation of the observed differences in viscosity between these xanthan-like polymers.     

Effect of misalignment on electrical characteristics of ACF joints for flip chip on flex applications

The effect of misalignment on the electrical properties of anisotropic conductive film (ACF) joints is investigated in this work. It is found that along with the increase of misalignment, the connection resistance of ACF joints increases. When the misalignment in x-direction is less than 5 μm, the increase rate of connection resistance is quite large. Then, along with the severity of misalignment, the increase rate becomes smaller. Finally, when the misalignment is close to 20 μm, the increase rate rises again. The Holm's electric contact theory is used for understanding the connection resistance variation. On the other hand, with the increase of misalignment in x-direction, the insulation resistance between ACF joints decreases. If the misalignment exceeded 10 μm, the decrease is prominent for the Ni particle ACF joints. This phenomenon can be explained by the effect of dielectric damage of the epoxy.Computer programs are also developed to calculate the variation of the probability of open and shorting after misalignment and predicate the maximum misalignment tolerance. The results show that the open and shorting probability increase abruptly after misalignment. On the view of pad parameters, the open probability is mainly related to the pad area, while the pads gap is critical to the shorting probability. Large pads gap (small pad width) can reduce the shorting probability obviously. On the other hand, enlarging the pad area by increasing pad length decreases the open probability significantly. So comparing to square shape pad, rectangle shape pad can reduce the failure probability greatly.     

High temperature controlled atmosphere experiments for catalysts in a transmission electron microscope

A new transmission electron microscopy (TEM) specimen preparation procedure for high temperature experiments using a controlled atmosphere specimen holder (HTCASH) has been developed. It is designed for studying the microstructure of catalyst specimens before and after treatments in various gases. The procedure involved (1) finding a new formula for the embedding material, (2) devising a new method of making specimen supports, and (3) developing a method for removing the embedding material after the specimen has been microtomed. These techniques were then brought together to produce the ideal specimens for the HTCASH experiments. As an extra benefit, this procedure is also suitable for preparing specimens for ultrahigh resolution imaging experiments. The application of the new procedure in HTCASH experiments is illustrated through a high temperature reduction of a Co/SiO₂-923 catalyst.     

Langmuir-Blodgett deposited cadmium gate inverted InP-GaInAs modulation-doped field-effect transistors

We report on the Langmuir-Blodgett film deposition and plasma etching of cadmium distearate on n-Gao.47Ino.53As to form a high-barrier-height Schottky barrier. Using this technique to form the gate electrode, we fabricated a 1?m-gate-length inverted InP-GalnAs modulation-doped field-effect transistor (MODFET) with an extrinsic transconductance of 170mS/mm and a cutoff frequencyfT of 19 GHz.     

Empirical comparison of image retrieval color similarity methods with human judgment

It is important to verify assumptions and methods of image retrieval against actual human behavior. A study was conducted to compare similarity methods of color histograms against human assessment of similarity. The similarity methods tested include basic histogram intersection, center histogram matching, locality histogram matching, and size-weighted histogram matching. 161 subjects participated in the empirical study. The findings, based on Spearman correlation analysis, showed that both the basic histogram intersection method and size-weighted histogram are very close to human assessment of similarity (Spearman correlation coefficient of 0.915). The other two are not close to human judgment on similarity. This study illustrates an alternative approach to evaluating matching algorithms. Unlike the usual measures of recall and precision, this approach emphasizes human validation. Fewer images are required with the use of statistical testing.     

Effect of refraction on optical microscopic measurement of internal blood-vessel diameter and its correction

In optical microscopic measurement of internal blood-vessel diameters, the effect of refraction must be taken into account to ensure accuracy of the result. This effect is discussed and an analytical correction formula derived. Phantom blood vessels with known internal and external diameters were used to test the validity of the correction formula. The errors obtained prior to correction were reduced significantly after correction.     

Investigation of the source/drain asymmetric effects due to gate misalignment in planar double-gate MOSFETs

A planar double-gate SOI MOSFET (DG-SOI) with thin channel and thick source/drain (S/D) was successfully fabricated. Using both experimental data and simulation results, the S/D asymmetric effect induced by gate misalignment was studied. For a misaligned DG-SOI, there is gate nonoverlapped region on one side and extra gate overlapped region on the other side. The nonoverlapped region introduces extra series resistance and weakly controlled channel, while the extra overlapped region introduces additional overlap capacitance and gate leakage current. We compared two cases: bottom gate shift to source side (DG/spl I.bar/S) and bottom gate shift to drain side (DG/spl I.bar/D). At the same gate misalignment value, DG/spl I.bar/S resulted in a larger drain-induced barrier lowering effect and smaller overlap capacitance at drain side than DG/spl I.bar/D. Because of reduced drain-side capacitance, the speed of three-stage ring oscillator of DG/spl I.bar/S, with 20% gate misalignment length (L/sub mis/) over gate length (L/sub g/), or L/sub mis//L/sub g/=20%, was faster than that of two-gate aligned DG-SOI.     

Axial-Field Electrical Machines - Design and Applications

Axial-field electrical machines offer an alternative to the conventional machines. In the axial-field machine, the airgap flux is axial in direction and the active current carrying conductors are radially positioned. This paper presents the design characteristics, special features, manufacturing aspects and potential applications for axial-field electrical machines. The experimental results from several prototypes, including d.c. machines, synchronous machines and single-phase machines are given. The special features of the axial-field machine, such as its planar and adjustable airgap, flat shape, ease of diversification, etc., enable axial-field machines to have distinct advantages over conventional machines in certain applications, especially in special-purpose applications.