Enzymatic Degradation of PrPSc by a Protease Secreted from Aeropyrum pernix K1

BACKGROUND: An R30 fraction from the growth medium of Aeropyrum pernix was analyzed for the protease that can digest the pathological prion protein isoform (PrP(Sc)) from different species (human, bovine, deer and mouse). METHODOLOGY/PRINCIPAL FINDINGS: Degradation of the PrP(Sc) isoform by the R30 fraction and the purified protease was evaluated using the 6H4 anti-PrP monoclonal antibody. Fragments from the N-terminal and C-terminal of PrP(Sc) were also monitored by Western blotting using the EB8 anti-PrP monoclonal antibody, and by dot blotting using the C7/5 anti-PrP monoclonal antibody, respectively. For detection of smaller peptides from incomplete digestion of PrP(Sc), the EB8 monoclonal antibody was used after precipitation with sodium phosphotungstate. Characterization of the purified active protease from the R30 fraction was achieved, through purification by fast protein liquid chromatography, and identification by tandem mass spectrometry the serine metalloprotease pernisine. SDS-PAGE and zymography show the purified pernisine plus its proregion with a molecular weight of ca. 45 kDa, and the mature purified pernisine as ca. 23 kDa. The purified pernisine was active between 58 °C and 99 °C, and between pH 3.5 and 8.0. The temperature and pH optima of the enzymatic activity of the purified pernisine in the presence of 1 mM CaCl(2) were 105 °C ± 0.5 °C and pH 6.5 ± 0.2, respectively. CONCLUSIONS/SIGNIFICANCE: Our study has identified and characterized pernisine as a thermostable serine metalloprotease that is secreted from A. pernix and that can digest the pathological prion protein PrP(Sc)

Interfacing war game simulations with tactical C2 systems - dream or reality?

Decision making process in current tactical C2 systems is base on planning process of commanders and their staff. Improving tactical decision making by interfacing war game simulations with tactical C2 systems is achievable. Commander can review the results of the simulation and subsequently modify the tactical plan. Previously, the use of "training" simulations was not a viable solution to real world decision making due to the lengthy time required to input all of the combat entities, the unit organizations and personnel dispositions, the equipment configurations, status of the units and equipment, and the distribution of the available supplies. Modern C2 systems have all of this information stored in the common system databases, and this information can be used to instantiate and populate the simulation through an electronic adaptation of the data structures to match the requirements of the constructive simulation. This paper will provide description of system approach of interfacing simulation and C2 system to improve decision-making

EUSpeech: A New Dataset of EU Elite Speeches

This paper presents EUSpeech, a new dataset of 18,403 speeches from EU leaders (i.e., heads of government in 10 member states, EU commissioners, party leaders in the European Parliament, and ECB and IMF leaders) from 2007 to 2015. These speeches vary in sentiment, topics and ideology, allowing for fine-grained, over-time comparison of representation in the EU