Structure-function relationships in membrane segment 5 of the yeast Pma1 H+-ATPase

Membrane segment 5 (M5) is thought to play a direct role in cation transport by the sarcoplasmic reticulum Ca2+-ATPase and the Na+, K+-ATPase of animal cells. In this study, we have examined M5 of the yeast plasma membrane H+-ATPase by alanine-scanning mutagenesis. Mutant enzymes were expressed behind an inducible heat-shock promoter in yeast secretory vesicles as described previously (Nakamoto, R. K., Rao, R., and Slayman, C. W. (1991) J. Biol. Chem. 266, 7940-7949). Three substitutions (R695A, H701A, and L706A) led to misfolding of the H+-ATPase as evidenced by extreme sensitivity to trypsin; the altered proteins were arrested in biogenesis, and the mutations behaved genetically as dominant lethals. The remaining mutants reached the secretory vesicles in sufficient amounts to be characterized in detail. One of them (Y691A) had no detectable ATPase activity and appeared, based on trypsinolysis in the presence and absence of ligands, to be blocked in the E1-to-E2 step of the reaction cycle. Alanine substitution at an adjacent position (V692A) had substantial ATPase activity (54%), but was likewise affected in the E1-to-E2 step, as evidenced by shifts in its apparent affinity for ATP, H+, and orthovanadate. Among the mutants that were sufficiently active to be assayed for ATP-dependent H+ transport by acridine orange fluorescence quenching, none showed an appreciable defect in the coupling of transport to ATP hydrolysis. The only residue for which the data pointed to a possible role in cation liganding was Ser-699, where removal of the hydroxyl group (S699A and S699C) led to a modest acid shift in the pH dependence of the ATPase. This change was substantially smaller than the 13-30-fold decrease in K+ affinity seen in corresponding mutants of the Na+, K+-ATPase (Arguello, J. M., and Lingrel, J. B (1995) J. Biol. Chem. 270, 22764-22771). Taken together, the results do not give firm evidence for a transport site in M5 of the yeast H+-ATPase, but indicate a critical role for this membrane segment in protein folding and in the conformational changes that accompany the reaction cycle. It is therefore worth noting that the mutationally sensitive residues lie along one face of a putative alpha-helix.     

The effect of partial substitution of nb for v on austenite grain size and hardness of a tool steel

In the present work, the composition of H- 13 tool steel was modified with respect to the vanadium content to include an addition of niobium to obtain improved austenite grain size control without altering the tempering response of the steel. Optical metallography and transmission electron microscopy (TEM) observation of extraction replicas showed that the new steel had a smaller austenite grain size and a finer carbide size distribution than the vanadium steel, which indicates a higher potential toughness.     

Hepatic damage produced by long-term alcohol consumption in well-nourished rats

A study of the effect of long-term alcohol consumption on the liver of well-nourished rats is described. Rats fed for 16 weeks on a semipurified diet supplemented with high levels of vitamins and lipotropic factors and alcohol corresponding to 35% of the total caloric intake developed marked fatty changes of the liver. Mild fatty changes were observed in pair-fed controls receiving as isoenergetic equivalent of sucrose instead of alcohol. Intracellular hyaline bodies, corresponding ultrastructurally to giant mitochondria were abundantly found in the hepatocytes of alcoholic rats, while in the controls they were not seen. Te findings in this investigation are postulated to provide further evidence that the long-term intake of alcohol exerts a direct causative role in the pathogenesis of liver damage.     

Cold Crystallization of PLLA Studied by Simultaneous SAXS and WAXS

Summary: The cold crystallization process of initially amorphous poly(L ‐lactic acid), PLLA, with two different molecular weights, during a heating at 2 °C/min, was investigated by DSC and time‐resolved simultaneous SAXS and WAXS, using synchrotron radiation. Equatorial scans of the isotropic 2D‐SAXS patterns showed that the average Bragg long period (L_B) of PLLA samples was approximately constant with the development of cold crystallization up to a temperature that corresponded to a melt/re‐crystallization process that took place before the nominal melting peak seen by DSC. L_B values were found to be higher for the high molecular weight material. This was in accordance with the higher melting temperature observed in the high molecular weight PLLA that implied the existence of thicker lamellae. WAXS results showed that the molecular weight did not apparently affect the crystal form and the final degree of crystallinity of PLLA. The Avrami parameters from WAXS and DSC were consistent, showing that the non‐isothermal cold crystallization of the two PLLA samples corresponded mainly to a three‐dimensional growth, although an imperfect crystallization process was involved at early times. The crystallization rate of PLLA, observed both by WAXS and DSC, decreased with increasing molecular weight.

SAXS profiles of PLLA2 as a function of temperature. The inset shows the 2D‐SAXS pattern obtained at 180 °C.     

Self-assembled monolayers of a disulphide-derivatised cobalt-porphyrin on gold

A self-assembled monolayer (SAM) of a novel cobalt(II)porphyrin disulphide derivative was prepared on flat gold(1 1 1) electrode. Evidence for surface modification was provided by electrochemical reductive desorption of the monolayer and ellipsometry, consistent with a coverage of 2.5 × 10⁻¹⁰ mol cm⁻² and a thickness of 13 Å, respectively. Both results support the presence of SAMs where the molecules share an intermediate position between perpendicular and flat orientation. Scanning tunnelling microscopy have also proven the formation of CoPSS SAMs, however high-resolution images could only be obtained when the CoPSS molecules were diluted in an hexanethiol SAM. The electrocatalytic activity of the surface confined Co-porphyrin was evaluated for the oxygen reduction. Voltammetric data indicate that reaction involves two electrons consistent with the formation of hydrogen peroxide. Under similar experimental conditions the data obtained for an iron-porphyrin analogue points for a full reduction of dioxygen to water.     

XMobile: A MB-UID environment for semi-automatic generation of adaptive applications for mobile devices

Ubiquitous Computing promises seamless access to information anytime, anywhere with different and heterogeneous devices. This kind of environment imposes new challenges to software development. For example, information and user interface should be adapted according to contextual characteristics such as user, environment, and access device. In case of device adaptation, the development challenge is related to the heterogeneity of the devices, which requires software engineers to create different versions for each type of device and every platform. This paper proposes a MB-UID (model-based user interface development) approach for semi-automatic generation of adaptive applications for mobile devices. An environment, called XMobile, offers a device-independent user interface framework and a code generation tool for providing fast development of multi-platform and adaptive applications according to device and platform features. A case study is also presented to illustrate how the environment can be used for constructing an application for heterogeneous devices with different network connectivity modes.     

Sliding Mode Control with Adaptive Fuzzy Dead-Zone Compensation of an Electro-hydraulic Servo-System

Electro-hydraulic servo-systems are widely employed in industrial applications such as robotic manipulators, active suspensions, precision machine tools and aerospace systems. They provide many advantages over electric motors, including high force to weight ratio, fast response time and compact size. However, precise control of electro-hydraulic systems, due to their inherent nonlinear characteristics, cannot be easily obtained with conventional linear controllers. Most flow control valves can also exhibit some hard nonlinearities such as dead-zone due to valve spool overlap. This work describes the development of an adaptive fuzzy sliding mode controller for an electro-hydraulic system with unknown dead-zone. The boundedness and convergence properties of the closed-loop signals are proven using Lyapunov stability theory and Barbalat’s lemma. Numerical results are presented in order to demonstrate the control system performance.     

An adaptive fuzzy sliding mode controller for remotely operated underwater vehicles

Sliding mode control is a very attractive control scheme because of its robustness against both structured and unstructured uncertainties as well as external disturbances. In this way, it has been widely employed for the dynamic positioning of remotely operated underwater vehicles. Nevertheless, in such situations the discontinuities in the control law must be smoothed out to avoid the undesirable chattering effects. The adoption of properly designed boundary layers has proven effective in completely eliminating chattering, however, leading to an inferior tracking performance. This work describes the development of a dynamic positioning system for remotely operated underwater vehicles. The adopted approach is primarily based on the sliding mode control strategy and enhanced by an adaptive fuzzy algorithm for uncertainty/disturbance compensation. Using the Lyapunov stability theory and Barbalat’s lemma, the boundedness and convergence properties of the closed-loop signals are analytically proven. The performance of the proposed control scheme is also evaluated by means of numerical simulations.     

Slicing the metric space to provide quick indexing of complex data in the main memory

Searching in a dataset for elements that are similar to a given query element is a core problem in applications that manage complex data, and has been aided by metric access methods (MAMs). A growing number of applications require indices that must be built faster and repeatedly, also providing faster response for similarity queries. The increase in the main memory capacity and its lowering costs also motivate using memory-based MAMs. In this paper, we propose the Onion-tree, a new and robust dynamic memory-based MAM that slices the metric space into disjoint subspaces to provide quick indexing of complex data. It introduces three major characteristics: (i) a partitioning method that controls the number of disjoint subspaces generated at each node; (ii) a replacement technique that can change the leaf node pivots in insertion operations; and (iii) range and k-NN extended query algorithms to support the new partitioning method, including a new visit order of the subspaces in k-NN queries. Performance tests with both real-world and synthetic datasets showed that the Onion-tree is very compact. Comparisons of the Onion-tree with the MM-tree and a memory-based version of the Slim-tree showed that the Onion-tree was always faster to build the index. The experiments also showed that the Onion-tree significantly improved range and k-NN query processing performance and was the most efficient MAM, followed by the MM-tree, which in turn outperformed the Slim-tree in almost all the tests.