Kinetic studies of a soluble alpha beta complex of nitrate reductase A from Escherichia coli. Use of various alpha beta mutants with altered beta subunits

A soluble alpha beta complex of nitrate reductase can be obtained from a strain of Escherichia coli that lacks the narI gene and expresses only the alpha and beta subunits. The beta subunit contains four Fe-S centres and the alpha subunit contains the molybdenum cofactor, which is the site at which nitrate is reduced. Despite the lack of the gamma subunit of the complete enzyme, this complex can still catalyse the reduction of nitrate with artificial electron donors such as benzyl viologen, so that it is suitable for studying the transfer of electrons between these two types of redox centre. To examine whether the electrons from reduced benzyl viologen are initially delivered to the Fe-S centres, or directly to the molybdenum cofactor, or both, we have studied the steady-state kinetics and the binding of benzyl viologen to the alpha beta complex and mutants alpha beta* with altered beta subunits. Reduction of the enzyme by reduced benzyl viologen in the absence of nitrate showed that all four Fe-S centres and the molybdenum cofactor could be reduced. Two classes of site with different equilibrium constants could be distinguished. The kinetic results suggest that benzyl viologen supplies its electrons directly to the molybdenum cofactor, at a rate showing a hyperbolic dependence on the square of the concentration of the electron donor. A reaction mechanism is proposed for the reduction of nitrate catalysed by the alpha beta complex of nitrate reductase with artificial electron donors.     

Extending the power line LAN up to the neighborhood transformer

This article reports on the performance of audio, video, multimedia, and other high-data-rate in-home networking applications. The article starts by describing the problems encountered in power line communication channels in terms of frequency response and noise characteristics, and explains how in-home power line LANs can be extended to the neighborhood transformer. OFDM physical layers providing speeds of 45 Mb/s and 200 Mb/s as well as QoS and security are introduced. Finally, the results of large tests involving several thousands of nodes are described.     

Sj?gren syndrome and subacute demyelinating polyradiculopathy: an unusual association

INTRODUCTION: Sj?gren's syndrome is a chronic inflammatory condition of unknown aetiology and autoimmune pathology. The defining feature is the dry syndrome, expressed as xerophthalmia and xerostomia. Extra-glandular involvement at many other levels may also occur. Neurological involvement is not unusual. The peripheral nervous system is most frequently involved, and a predominantly sensitive symmetrical distal polyneuropathy may be the first sign of the condition. Other patterns of peripheral involvement are also associated with the syndrome. We present a case of subacute demyelinating polyradiculopathy associated with primary Sj?gren's syndrome. CLINICAL CASE: A 28 year old woman with dry syndrome presented with paraesthesia in her hands and feet, distal weakness, which had progressed proximally in the muscles of her arms and legs, and bilateral facial weakness. The condition progressed for eight weeks. When complementary tests were done, alterations typical of this condition (FR, ANA, anti-Ro and anti-La) were seen and also others typical of the dry syndrome (Schirmer's test). Therefore, in view of these findings and the clinical features, after other conditions had been ruled out, a diagnosis of primary Sj?gren's syndrome was made. The type of neuropathy was determined by the clinical features, electromyography and CSF findings. Treatment with corticosteroids gave good results. CONCLUSIONS: Demyelinating polyradiculopathy is a form of peripheral nervous system involvement which is rarely seen in this disorder. In the differential diagnosis Sj?gren's syndrome should be considered, an orientative history taken, autoantibodies determined and an ophthalmological examination made.     

A framework for avoiding steganography usage over HTTP

Steganographic techniques allow users to covertly transmit information, hiding the existence of the communication itself. These can be used in several scenarios ranging from evading censorship to discreetly extracting sensitive information from an organization. In this paper, we consider the problem of using steganography through a widely used network protocol (i.e. HTTP). We analyze the steganographic possibilities of HTTP, and propose an active warden model to hinder the usage of covert communication channels. Our framework is meant to be useful in many scenarios. It could be employed to ensure that malicious insiders are not able to use steganography to leak information outside an organization. Furthermore, our model could be used by web servers administrators to ensure that their services are not being abused, for example, as anonymous steganographic mailboxes. Our experiments show that steganographic contents can be successfully eliminated, but that dealing with high payload carriers such as large images may introduce notable delays in the communication process.     

Socio-economic indicators on research and development in Spain

This essay analyses some aspects of the situation of scientific policy in Spain, mainly from a socio-economic point of view. The funds dedicated to R and D, its sources, evolution by years and distribution in relation with the GNP and different sectors such as public institutions and private enterprises are studied, as well as the relations of those funds with the inhabitants of the country and with the number of scientific researchers, the areas of research and the percentages dedicated to applied research and to development. In a similar way personnel engaged in R and D, their number, types of researchers, centers and scientific areas of research are presented.This paper was in part the author's contribution to the Seminar on Evaluation in the Management of R and D, organized by the Economic Comission for Europe-United Nations. Madrid. Spain. April, 1989.     

A new graphical visualization of n-dimensional Pareto front for decision-making in multiobjective optimization

New challenges in engineering design lead to multiobjective (multicriteria) problems. In this context, the Pareto front supplies a set of solutions where the designer (decision-maker) has to look for the best choice according to his preferences. Visualization techniques often play a key role in helping decision-makers, but they have important restrictions for more than two-dimensional Pareto fronts. In this work, a new graphical representation, called Level Diagrams, for n-dimensional Pareto front analysis is proposed. Level Diagrams consists of representing each objective and design parameter on separate diagrams. This new technique is based on two key points: classification of Pareto front points according to their proximity to ideal points measured with a specific norm of normalized objectives (several norms can be used); and synchronization of objective and parameter diagrams. Some of the new possibilities for analyzing Pareto fronts are shown. Additionally, in order to introduce designer preferences, Level Diagrams can be coloured, so establishing a visual representation of preferences that can help the decision-maker. Finally, an example of a robust control design is presented - a benchmark proposed at the American Control Conference. This design is set as a six-dimensional multiobjective problem.     

User simulation in a stochastic dialog system

We present a new methodology of user simulation applied to the evaluation and refinement of stochastic dialog systems. Common weaknesses of these systems are the scarceness of the training corpus and the cost of an evaluation made by real users. We have considered the user simulation technique as an alternative way of testing and improving our dialog system. We have developed a new dialog manager that plays the role of the user. This user dialog manager incorporates several knowledge sources, combining statistical and heuristic information in order to define its dialog strategy. Once the user simulator is integrated into the dialog system, it is possible to enhance the dialog models by an automatic strategy learning. We have performed an extensive evaluation, achieving a slight but clear improvement of the dialog system.     

Covalent Adaptable Microstructures via Combining Two-Photon Laser Printing and Alkoxyamine Chemistry: Toward Living 3D Microstructures

Manufacturing programmable materials, whose mechanical properties can be adapted on demand, is highly desired for their application in areas ranging from robotics, to biomedicine, or microfluidics. Herein, the inclusion of dynamic and living bonds, such as alkoxyamines, in a printable formulation suitable for two-photon 3D laser printing is exploited. On one hand, taking advantage of the dynamic covalent character of alkoxyamines, the nitroxide exchange reaction is investigated. As a consequence, a reduction of the Young´s Modulus by 50%, is measured by nanoindentation. On the other hand, due to its “living” characteristic, the chain extension becomes possible via nitroxide mediated polymerization. In particular, living nitroxide mediated polymerization of styrene results not only in a dramatic increase of the volume (≈8 times) of the 3D printed microstructure but also an increase of the Young's Modulus by two orders of magnitude (from 14 MPa to 2.7 GPa), while maintaining the shape including fine structural details. Thus, the approach introduces a new dimension by enabling to create microstructures with dynamically tunable size and mechanical properties.     

Technological Implications of Modifying the Extent of Cell Wall-Proanthocyanidin Interactions Using Enzymes

The transference and reactivity of proanthocyanidins is an important issue that affects the technological processing of some fruits, such as grapes and apples. These processes are affected by proanthocyanidins bound to cell wall polysaccharides, which are present in high concentrations during the processing of the fruits. Therefore, the effective extraction of proanthocyanidins from fruits to their juices or derived products will depend on the ability to manage these associations, and, in this respect, enzymes that degrade these polysaccharides could play an important role. The main objective of this work was to test the role of pure hydrolytic enzymes (polygalacturonase and cellulose) and a commercial enzyme containing these two activities on the extent of proanthocyanidin-cell wall interactions. The results showed that the modification promoted by enzymes reduced the amount of proanthocyanidins adsorbed to cell walls since they contributed to the degradation and release of the cell wall polysaccharides, which diffused into the model solution. Some of these released polysaccharides also presented some reactivity towards the proanthocyanidins present in a model solution.     

Grape seed removal: effect on phenolics,chromatic and organoleptic characteristics of red wine

Winemakers are interested in wines with low bitterness and astringency, and therefore, there is a preference for a higher presence of skin proanthocyanidins in wines. However, the information regarding the effect of reducing or eliminating seed proanthocyanidins from the must or fermenting wine on the characteristics of finished wines is scarce. For this reason, we elaborated a wine using Monastrell grapes from which the seeds had been removed and compared it with a control wine (CW) made from intact grapes. The anthocyanin and proanthocyanidin concentration and profile, and the chromatic and sensory characteristics were studied. The absence of seed proanthocyanidins did not affect anthocyanin concentration, although the proanthocyanidin concentration was 40% lower than in the CW. The copigmentation (CA) study showed that both wines had a similar extent of CA phenomena although colour intensity was higher when seed proanthocyanidins were present. The panellist evaluated the wines elaborated without seeds as being fruitier, less astringent and with an overall higher quality.