Verification and Validation Test Plan for Prague Ruzyne Airport

This document is positioned within the framework of activities for the ´European airport Movement Management by A-SMGCS (EMMA)´ Project. Sub-project SP6 deals with the Verification and Validation (V and V) activities to be carried out within the Project. This document is the deliverable D6.1.2, the V and V Test Plan for the A-SMGCS at Prague-Ruzyne Airport. Its purpose is to: 1. Identify the V and V aims, objectives and hypotheses for the tests 2. Plan and prepare the validation exercises 3. Provide a high-level description of how the V and V activities will be conducted

Test Results Prague

The document is the test report of the Prague test campaign that were conducted in the Integrated Project EMMA, a project of the sixth framework programme of the European Commission. It contains all results and conclusion that were gained with real time simulations and technical and operational trials at Prague Airport

Validation Test Plan PRG

The test plans for two real-time simulations and a field test are documented here. Higher A-SMGCS services were tested in the Prague Airport environment. Aims and objectives, experimental factors and the experiment design are descripted here. Furthermore, the necessary metrics and measurements, the hypotheses that can be accepted or rejected based on the measurements made, and the complete test environment. In addition to that scenario specifications are given and requirements for participants and training of participants are determined. Finally, the conduct of the experiments is described and the envisioned analysis methods is outlined. The document follows the E-OCVM guidelines

Crystallographic Studies on the Ribosome, a Large Macromolecular Assembly Exhibiting Severe Nonisomorphism, Extreme Beam Sensitivity and No Internal Symmetry

Crystals, diffracting best to around 3 Å, have been grown from intact large and small ribosomal subunits. The bright synchrotron radiation necessary for the collection of the higher-resolution X-ray diffraction data introduces significant decay even at cryo temperatures. Nevertheless, owing to the reasonable isomorphism of the recently improved crystals of the small ribosomal subunits, reliable phases have been extracted at medium resolution (5-6 Å) and an interpretable five-derivative MIR map has been constructed. For the crystals of the large subunits, however, the situation is more complicated because at higher resolution (2.7-7 Å) they suffer from substantial radiation sensitivity, a low level of isomorphism, instability of the longest unit-cell axis and nonisotropic mosaicity. The 8 Å MIR map, constructed to gain insight into this unusual system, may provide feasible reasoning for the odd combination of the properties of these crystals as well as hints for future improvement. Parallel efforts, in which electron-microscopy-reconstructed images are being exploited for molecular-replacement studies, are also discussed

A milestone in ribosomal crystallography: the construction of preliminary electron density maps at intermediate resolution

Preliminary electron density maps of the large and the small ribosomal particles from halophilic and thermophilic sources, phased by the isomorphous replacement method, have been constructed at intermediate resolution. These maps contain features comparable in size with what is expected for the corresponding particles, and their packing arrangements are in accord with the schemes obtained by ab-initio procedures as well as with the motifs observed in thin sections of the crystals by electron microscopy. To phase higher resolution data, procedures are being developed for derivatization by specific labeling of the ribosomal particles at selected locations with rather small and dense clusters. Potential binding sites are being inserted either by site directed mutagenesis or by chemical modifications to facilitate cluster binding on the surface of the halophilic large and the thermophilic small ribosomal particles, which yield the crystals diffracting to highest resolution (2.9 and 7.3 Å (1 Å = 0.1 nm), respectively). For this purpose, the surface of these ribosomal particles is being characterized and procedures are being developed for quantitative detachment of selected ribosomal proteins and for their incorporation into core particles. The genes of these proteins are being cloned, sequenced, mutated to introduce reactive side groups, mainly cysteines, and overexpressed. In parallel, two in situ small and stable complexes were isolated from the halophilic ribosome. Procedures for their crystal production in large quantities are currently being developed. Models, reconstructed at low resolution from crystalline arrays of ribosomes and their large subunits, are being used for initial low-resolution phasing of the X-ray amplitudes. The interpretation of these models stimulated the design and the crystallization of complexes mimicking defined functional states of a higher quality than those obtained for isolated ribosomes. These models also inspired modelling experiments according to results of functional studies, performed elsewhere, focusing on the progression of nascent proteins.Key words: ribosomes, crystallography, undecagold cluster, heteropolyanions