Fluorescence Dequenching Makes Haem-Free Soluble Guanylate Cyclase Detectable in Living Cells

In cardiovascular disease, the protective NO/sGC/cGMP signalling-pathway is impaired due to a decreased pool of NO-sensitive haem-containing sGC accompanied by a reciprocal increase in NO-insensitive haem-free sGC. However, no direct method to detect cellular haem-free sGC other than its activation by the new therapeutic class of haem mimetics, such as BAY 58-2667, is available. Here we show that fluorescence dequenching, based on the interaction of the optical active prosthetic haem group and the attached biarsenical fluorophor FlAsH can be used to detect changes in cellular sGC haem status. The partly overlap of the emission spectrum of haem and FlAsH allows energy transfer from the fluorophore to the haem which reduces the intensity of FlAsH fluorescence. Loss of the prosthetic group, e.g. by oxidative stress or by replacement with the haem mimetic BAY 58-2667, prevented the energy transfer resulting in increased fluorescence. Haem loss was corroborated by an observed decrease in NO-induced sGC activity, reduced sGC protein levels, and an increased effect of BAY 58-2667. The use of a haem-free sGC mutant and a biarsenical dye that was not quenched by haem as controls further validated that the increase in fluorescence was due to the loss of the prosthetic haem group. The present approach is based on the cellular expression of an engineered sGC variant limiting is applicability to recombinant expression systems. Nevertheless, it allows to monitor sGC's redox regulation in living cells and future enhancements might be able to extend this approach to in vivo conditions

Simulation of the diffusion features of point defects in bcc metals

This work is devoted to simulation of the diffusion features of point defects in bee metals. The properties of point defects have been investigated with the usage of many-body interatomic potentials. This approach, based on the density-functional theory, permitted us to derive more adequate diffusion features of solids. This investigation is carried out within the framework of the Finnis-Sinclair formalism, developed for an assembly of N atoms and represents the second-moment approximation of the tight-binding theory. We used a new model, based on the molecular static method for simulating the atomic structure near the defect and vacancy migration in pure metals. This approach gives the opportunity to simulate the formation and the migration volumes of the point defects, taking into consideration the influence of pressure on structure and consequently on energy. The diffusion characteristics of bee alpha-Fe and anomalous beta-Zr have been investigated.</p

Simulation of the diffusion features of point defects in bcc metals

This work is devoted to simulation of the diffusion features of point defects in bee metals. The properties of point defects have been investigated with the usage of many-body interatomic potentials. This approach, based on the density-functional theory, permitted us to derive more adequate diffusion features of solids. This investigation is carried out within the framework of the Finnis-Sinclair formalism, developed for an assembly of N atoms and represents the second-moment approximation of the tight-binding theory. We used a new model, based on the molecular static method for simulating the atomic structure near the defect and vacancy migration in pure metals. This approach gives the opportunity to simulate the formation and the migration volumes of the point defects, taking into consideration the influence of pressure on structure and consequently on energy. The diffusion characteristics of bee alpha-Fe and anomalous beta-Zr have been investigated

Experimental Results on a 1.5??MW, 110??GHz Gyrotron with a Smooth Mirror Mode Converter

We present an internal mode converter (IMC) design for a 1.5 MW, 110 GHz gyrotron operating in the TE(22,6) mode. The launcher, designed using the codes Surf3d and LOT, converts the cavity waveguide mode into a nearly pure Gaussian beam. The Gaussian beam output from the launcher is shaped by a series of 4 smooth, curved mirrors to provide a circular output beam with a flat phase front at the gyrotron window. By employing smooth mirrors rather than mirrors with phase correcting surfaces, such an IMC is less sensitive to alignment issues and can more reliably operate with high efficiency. The IMC performance was verified by both cold test and hot test experiments. Beam pattern measurements in each case were in good agreement with theoretical predictions. The output beam was of high quality with calculations showing that the Gaussian Beam content was 95.8 +/- 0.5% in both hot and cold test