Novalis frameless image-guided noninvasive radiosurgery: initial experience.

OBJECTIVE: To evaluate our initial experience with Novalis (BrainLAB, Heimstetten, Germany) frameless image-guided noninvasive radiosurgery. METHODS: The system combines the dedicated Novalis linear accelerator with ExacTrac X-Ray 6D, an infrared camera and a kilovolt stereoscopic x-ray imaging system, a noninvasive mask system, and ExacTrac robotics for patient positioning in six degrees of freedom. Reference cranial skeletal structures are radiographically imaged and automatically fused to digital reconstructed radiographs calculated from the treatment planning computed tomographic scan to find the target position and accomplish automatic real-time tracking before and during radiosurgery. We present the acceptance testing and initial experience in 15 patients with 19 intracranial lesions treated between December 2005 and June 2006 at the Charité by frameless image-guided radiosurgery with doses between 12 and 20 Gy prescribed to the target-encompassing isodose. RESULTS: Phantom tests showed an overall system accuracy of 1.04 +/- 0.47 mm, with an average in-plane deviation of 0.02 +/- 0.96 mm for the x-axis and 0.02 +/- 0.70 mm for the y-axis. After infrared-guided patient setup of all patients, the overall average translational deviation determined by stereoscopic x-ray verification was 1.5 +/- 1.3 mm, and the overall average rotational deviation was 1.0 +/- 0.8 degree. The data used for radiosurgery, after stereoscopic x-ray verification and correction, demonstrated an overall average setup error of 0.31 +/- 0.26 mm for translation and 0.26 +/- 0.23 degree for rotation. CONCLUSION: This initial evaluation demonstrates the system accuracy and feasibility of Novalis image-guided noninvasive radiosurgery for intracranial benign and malignant lesions.

Formation, TEM study and 3D reconstruction of the human erythrocyte peroxiredoxin-2 dodecahedral higher-order assembly.

The production of a higher-order assembly of peroxiredoxin-2 (Prx-2) from human erythrocytes has been achieved during specimen preparation on holey carbon support films, in the presence of ammonium molybdate and polyethylene glycol. TEM study suggested that this assembly is a regular dodecahedron, containing 12 Prx-2 decamers (Mr 2.62 MDa, external diameter approximately 20 nm). This interpretation has been supported by production of a approximately 1.6 nm 3D reconstruction from the negative stain TEM data, with automated docking of the available X-ray data of the Prx-2 decamer. Comparison with other known protein dodecahedral and viral icosahedral structures indicates that this arrangement of protein molecules is one of the fundamental macromolecular higher-order assemblies found in biology. Widespread biotechnological interest in macromolecular "cage" structures is relevant to the production of the Prx-2 dodecahedron.

Interaction of the Vibrio cholerae cytolysin (VCC) with cholesterol, some cholesterol esters, and cholesterol derivatives: a TEM study.

The Vibrio cholerae cytolysin (VCC) 63-kDa monomer has been shown to interact in aqueous suspension with cholesterol microcystals to produce a ring/pore-like heptameric oligomer approximately 8 nm in outer diameter. Transmission electron microscopy data were produced from cholesterol samples adsorbed to carbon support films, spread across the holes of holey carbon films, and negatively stained with ammonium molybdate. The VCC oligomers initially attach to the edge of the stacked cholesterol bilayers and with increasing time cover the two planar surfaces. VCC oligomers are also released into solution, with some tendency to cluster, possibly via the hydrophobic membrane-spanning domain. At the air/water interface, the VCC oligomers are likely to be selectively oriented with the hydrophobic domain facing the air. Despite some molecular disorder/plasticity within the oligomers, multivariate statistical analysis and rotational self-correlation using IMAGIC-5 strongly suggest the presence of sevenfold rotational symmetry. To correlate the electron microscopy data with on-going biochemical and permeability studies using liposomes of varying lipid composition, the direct interaction of VCC with several cholesterol derivatives and other steroids has been examined. 19-Hydroxycholesterol and 7 beta-hydroxycholesterol both induce VCC oligomerization. beta-Estradiol, which does not possess an aliphatic side chain, also efficiently induces VCC oligomer formation, as does cholesteryl acetate. Cholesteryl stearate and oleate and the C22 (2-trifluoroacetyl)naphthyloxy analogue of cholesterol fail to induce VCC oligomerization, but binding of the monomer to the surface of these steroids does occur. Stigmasterol has little tendency to induce oligomer formation, and oligomers are largely confined to the edge of the bilayers; ergosterol has even less oligomerization ability. Attempts to solubilize and stabilize the VCC oligomers from cholesterol suspensions have been pursued using the neutral surfactant octylglucoside. Although individual solubilized oligomers have been defined which exhibit a characteristic cytolysin channel conformation in the side-on orientation, a tendency remains for the oligomers to cluster via their hydrophobic domains.

Routine preparation of air-dried negatively stained and unstained specimens on holey carbon support films: a review of applications.

Several representative examples are given of the successful application of negative staining across the holes of holey carbon support films using 5% (w/v) ammonium molybdate solution containing trehalose. The inclusion of 0.1% (w/v) trehalose is considered to be most satisfactory, although good data have also been obtained in the presence of 0.01 and 1.0% (w/v) trehalose. The examples given fall into the following groups: protein molecules in the absence of polyethylene glycol (PEG), protein molecules in the presence of PEG (Mr 1000), lipoproteins, lipids and membranes, filaments and tubules, viruses in the absence of PEG, viruses in the presence of PEG, aqueous polymer solutions, and finally for comparison purposes, four unstained samples studied in the presence of trehalose alone. In all these cases, and many others not documented here, successful spreading of the sample across holes has been achieved, with the sample embedded within a thin film of air-dried ammonium molybdate+trehalose. These specimens can be rapidly produced and provide an alternative to negatively stained specimens on carbon support films. Specimen stability in the electron bean is good and such specimens can usually generate superior negatively stained TEM images without flattening and adsorption artefacts. The formation of 2-D arrays/crystals of protein molecules and viruses, suspended across holes in the presence of ammonium molbybdate+trehalose, and trehalose alone, is also demonstrated.