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.     

Experimental Investigation of the Recovery of Soaked Paper Using Evaporative Freeze Drying

The aim of this article is to develop an experiment and a procedure to investigate the restoration of water-damaged paper and archival materials using freeze drying in order to allow a reproducible test and comparison of the influence of different operating conditions on drying time and restored paper quality. Firstly, a reproducible method for the preparation of soaked samples simulating water-damaged paper has been developed. Then, the samples have been freeze-dried in a laboratory-scale apparatus that allowed monitoring the temperature as well as the weight of the samples. The technique of evaporative freezing, which reduces the drying time required, has been used in this case. An innovative procedure for the visualization of the progress of the drying process has been validated, thus allowing the validation of a simple phenomenological model of the time evolution of the ice core volume; in addition, data on the residual moisture of the dried paper sheets in different zones have been given. Finally, optimization of this particular drying process by using simple or more sophisticated approaches has been discussed.     

Proteomic profile of differentially expressed plasma proteins from dystrophic mice and following suberoylanilide hydroxamic acid treatment

Purpose: Histone Deacetylase Inhibitors (DI) ameliorates dystrophic muscle regeneration restoring muscular strength in the mdx mouse model of Duchenne muscular dystrophy (DMD). The further development of these compounds as drugs for DMD treatment is currently hampered by the lack of knowledge about DIs effect in large dystrophic animal models and that of suitable biomarkers to monitor their efficacy. Experimental design: In this study we applied proteomic analysis to identify differentially expressed proteins present in plasma samples from mdx mice treated with the Suberoylanilide hydroxamic acid (SAHA) and relative normal controls (WT). Results: Several differentially expressed proteins were identified between untreated wild type and mdx mice. Among these, fibrinogen, epidermal growth factor 2 receptor, major urinary protein and glutathione peroxidase 3 (GPX3) were constitutively up‐regulated in mdx, while complement C3, complement C6, gelsolin, leukaemia inhibitory factor receptor (LIFr), and alpha 2 macroglobulin were down‐regulated compared to WT mice. SAHA determined the normalization of LIFr and GPX3 protein level while apoliprotein E was de novo up‐regulated in comparison to vehicle‐treated mdx mice. Conclusions and clinical relevance: Collectively, these data unravel potential serological disease biomarkers of mdx that could be useful to monitor muscular dystrophy response to DI treatment.     

Traffic analysis and synthetic models of second life

Virtual Worlds (VW), such as Massive Multiplayer Online Social Games, have been gaining increasing attention in the last few years, mainly due to the new way users interact with them. However, little effort has been devoted to understand their traffic profile and the implications to the traffic management area. With the current growing rate of VWs’ usage, their traffic demand could eventually impose a significant burden on the operation of a typical Internet Service Provider (ISP) network. In this paper, we seek to understand the traffic behavior of an increasingly popular VW application, namely Second Life (SL), from both the connection and network level perspectives. We also show results of a traffic analysis of a SL client, when an avatar performs different actions in the VW, at different places and under different network conditions. Our results show that SL makes intensive use of network resources (mostly bandwidth), since the capacity needed for having a full SL experience (listening to live music) may reach 500 kbps. We then extend the traffic analysis work on SL by providing analytical models for its traffic profile. We aim at helping the network management and planning area to estimate the impact of an intense use of such VW on access links or core networks. Such synthetic models are also useful to the networking research community for the use in network simulations.     

Emergent semantics through interaction in image databases

In this paper, we briefly discuss some aspects of image semantics and the role that it plays for the design of image databases. We argue that images don't have an intrinsic meaning, but that they are endowed with a meaning by placing them in the context of other images and by the user interaction. From this observation, we conclude that, in an image, database users should be allowed to manipulate not only the individual images, but also the relation between them. We present an interface model based on the manipulation of configurations of images     

The behavior of the4He viscosity near the superfluid transition

Measurements of the shear viscosity at saturated vapor pressure through the lambda transition indicate a singular behavior of the form |1 – (/)|=A ^x , (where =|1–(T/T )|, with equal values for the critical exponent on both sides of the transition.Work sponsored by Consiglio Nazionale delle Ricerche, Rome (Italy).     

Neural networks based subgrid scale modeling in large eddy simulations

In this paper a multilayer feed-forward neural network (NN) is used as subgrid scale (SGS) model in a large eddy simulation (LES). The NN was previously off-line trained using numerical data generated by a LES of a channel flow at Re_τ=180 with Bardina's scale similar (BFR) SGS model. Results show the ability of NNs to identify and reproduce the highly nonlinear behavior of the turbulent flows, and therefore the possibility of using NN techniques in numerical simulations of turbulent flows.