A column centrifugation method for the reconstitution in liposomes of the mitochondrial F0F1 ATP synthase/ATPase

A method for reconstitution of membrane proteins into unilamellar liposomes is described. The model enzyme was the F0F1 ATP synthase from mitochondria when in complex or free from its inhibitor protein. The enzymes were first solubilized with either of two detergents, i.e., n-dodecyl-beta-D maltoside or lauryldimethylamine oxide. After solubilization, the enzymes were passed through a column of Sepharose-AH using an ADP/sodium cholate selective elution buffer. The enzymes recovered from the column were subsequently passed through a centrifuge column of Sephadex G-50 fine. The eluate contained liposomes in which the F0F1 complex (with and without inhibitor protein) had been reconstituted. The reconstituted enzymes were capable of hydrolyzing ATP with formation of electrochemical H+ gradients. They also catalyzed the ATP-Pi exchange reactions. Thus the F0F1 complex which is formed by 18 subunits can be rapidly reconstituted into liposomes in a fully functional state. Moreover the data show that the interactions between the enzyme and its inhibitor protein are not perturbed in the reconstitution procedure.     

Fracture of glass in tensile and bending tests

To fully characterize the mechanical behavior of ceramics, it is useful to study their response to both tensile and bending tests. In this investigation, a quantitative comparison is made between tensile and bending results from (circularly) cylindrical glass rods of borosilicate glass and of fused silica. These experimental results show that in each material, the Weibull exponentm is approximately the same for both types of test. The fractured samples clearly showed that all failures initiated at surface-located flaws. The stress level at which the probability of fracture in tension equals 50 Pct was determined; the maximum tensile stress for 50 Pct probability of fracture in bending was also determined. The ratio of these two stresses was predicted previously from simple, theoretical considerations based upon the applicability of the Weibull function and on the distribution of defects in the specimens. This theoretical result is inconsistent with the experimental observations in view of the fractography. Therefore, it is concluded that failure in the materials tested is not satisfactorily described by Weibull statistics.     

Properties of permease dimer, a fusion protein containing two lactose permease molecules from Escherichia coli

An engineered fusion protein containing two tandem lactose permease molecules (permease dimer) exhibits high transport activity and is used to test the phenomenon of negative dominance. Introduction of the mutation Glu-325-->Cys into either the first or the second half of the dimer results in a 50% decrease in activity, whereas introduction of the mutation into both halves of the dimer abolishes transport. Lactose transport by permease dimer is completely inactivated by N-ethylmaleimide; however, 40-45% activity is retained after N-ethylmaleimide treatment when either the first or the second half of the dimer is replaced with a mutant devoid of cysteine residues. The observations demonstrate that both halves of the fusion protein are equally active and suggest that each half may function independently. To test the possibility that oligomerization between dimers might account for the findings, a permease dimer was constructed that contains two different deletion mutants that complement functionally when expressed as untethered molecules. Because this construct does not catalyze lactose transport to any extent whatsoever, it is unlikely that the two halves of the dimer interact or that there is an oligomeric interaction between dimers. The approach is consistent with the contention that the functional unit of lactose permease is a monomer.     

Solution of temperature fields in hydrodynamics bearings by the numerical network method

Solution to the 2-D steady-state nonlinear heat conduction equation, involving cylindrical coordinates, applied to a plain bearing by a simple and versatile numerical technique based on network method is presented in this work. These advantages of the technique are necessary for the status evaluation of industrial machines during operation because time and computing resources are limited. The pressure field in the fluid is previously solved from the Reynolds equation by finite element method. The main difficulty is associated with the exponential dependency of the viscosity on temperature. The proposed model is very efficient and requires negligible computing times. Solutions are very close to the experimental and numerical results of other researchers.     

Ultrasound‐activated Knoevenagel condensation of malononitrile with carbonylic compounds catalysed by alkaline‐doped saponites

α,β‐Unsaturated nitriles have been synthesized by Knoevenagel condensation of a carbonylic compound with malononitrile, assisted by sonochemical irradiation. Two alkaline‐promoted clays (Li⁺‐ and Cs⁺‐exchanged saponites) have been employed as catalysts. The influence of the carbonylic compound (benzaldehyde or cyclohexanone) and the use of a solvent on the catalytic activity have been studied. Remarkable increase in the conversion values has been found when the reaction is activated by ultrasound, as compared with the thermal activation. In this green, solvent‐free procedure, α,β‐unsaturated nitriles have been produced in very high yields (97%) when the Cs⁺‐saponite is used as catalyst. Copyright © 2004 Society of Chemical Industry     

Transmission Fiber Chromatic Dispersion Dependence on Temperature: Implications on 40 Gb/s Performance

In this letter, we will evaluate the performance degradation of a 40 km high‐speed (40 Gb/s) optical system, induced by optical fiber variations of the chromatic dispersion induced by temperature changes. The chromatic dispersion temperature sensitivity will be estimated based on the signal quality parameters.     

Tocopherol contents of some Turkish wine by-products

The aim of this study was to determine the tocopherol contents of wine by-products including grape seed, pomace (seed, skin and stem) and bagasse (skin and stem) by using two different extraction methods in order to evaluate the possibilities of them as natural non-polar antioxidants. Cold extraction appeared to give more tocopherol contents in seeds than hot extraction. But hot extraction was found more suitable for pomace and bagasse. α-Tocopherol was the most abundant tocopherol in the grape seed, pomace and bagasse. Although γ- and δ-tocopherols were found in the samples with low concentrations, β-tocopherol was not detected in the samples. Kalecik karası had the highest α and total tocopherol compared to the other cultivars. On the other hand, hot extracted bagasse in all cultivars gave the highest tocopherol contents. As a conclusion, wine by-products, a large scale wastes, can be evaluated both to get natural tocopherol source and to obtain economical gain.     

Model Driven Architecture

Bei der Model Driven Architecture (MDA) bilden Modelle die zentralen Elemente des Softwareentwicklungsprozesses. Ziel ist es, plattformspezifische Modelle möglichst automatisiert aus plattformunabhängigen Modellen abzuleiten. Dadurch soll der Aufwand der Softwareentwicklung verringert und die Adaptierung an neue Technologien erleichtert werden.*Vorschläge an Prof. Dr. Frank Puppe oder Dieter Steinbauer Alle „Aktuellen Schlagwörter“ seit 1988 finden Sie unter: www.ai-wuerzburg.de/as     

Disassembly sequence planning in a disassembly cell context

In this paper a two-phase approach is proposed for determining the optimal disassembly sequence when the disassembly system has a cellular configuration. Operations are first grouped into cells based on the resources they require with the goal of minimizing machine acquisition costs. The aim is to group together those operations that use similar equipment in order to achieve good utilization levels of such equipment. A maximum cell size may be imposed. Once the cells have been formed and the operations have been assigned to them, a metaheuristic algorithm (namely GRASP) is used to search for the disassembly sequence for each product that leads to the minimum number of intercellular movements. To account for uncertainty regarding the condition in which the product may arrive, each disassembly task is assumed to be required with a certain probability, regardless of the other tasks. AND/OR precedence relations among the disassembly tasks are also considered. The proposed approach is illustrated on a randomly generated disassembly problem.