Expression of Staphylococcus aureus clumping factor A in Lactococcus lactis subsp. cremoris using a new shuttle vector.

Staphylococcus aureus harbors redundant adhesins mediating tissue colonization and infection. To evaluate their intrinsic role outside of the staphylococcal background, a system was designed to express them in Lactococcus lactis subsp. cremoris 1363. This bacterium is devoid of virulence factors and has a known genetic background. A new Escherichia coli-L. lactis shuttle and expression vector was constructed for this purpose. First, the high-copy-number lactococcal plasmid pIL253 was equipped with the oriColE1 origin, generating pOri253 that could replicate in E. coli. Second, the lactococcal promoters P23 or P59 were inserted at one end of the pOri253 multicloning site. Gene expression was assessed by a luciferase reporter system. The plasmid carrying P23 (named pOri23) expressed luciferase constitutively at a level 10,000 times greater than did the P59-containing plasmid. Transcription was absent in E. coli. The staphylococcal clumping factor A (clfA) gene was cloned into pOri23 and used as a model system. Lactococci carrying pOri23-clfA produced an unaltered and functional 130-kDa ClfA protein attached to their cell walls. This was indicated both by the presence of the protein in Western blots of solubilized cell walls and by the ability of ClfA-positive lactococci to clump in the presence of plasma. ClfA-positive lactococci had clumping titers (titer of 4,112) similar to those of S. aureus Newman in soluble fibrinogen and bound equally well to solid-phase fibrinogen. These experiments provide a new way to study individual staphylococcal pathogenic factors and might complement both classical knockout mutagenesis and modern in vivo expression technology and signature tag mutagenesis.

Economic impact of a new museum on the local economy: "The Gaudí Centre"

In this paper we analyse the economic impact of a new museum (the Gaudí Centre) on the local economy of Reus, a city in the province of Tarragona (southern Catalonia). We use a Keynesian income multiplier model to evaluate the effects of this new cultural venue on local income. In our calculation of the economic impact we distinguish between two phases: the construction phase and the exploitation phase. Our results show the important income impact of this cultural investment on the local economy.

Olympic Games as showcase

In the 1992 Barcelona Olympic Games, besides the country-versus-country typical showdown in the backdrop of Olympics, and the economic and political impulses for the host city, there was a third factor complicating the celebration: rivalry between the Catalan hosts and the Spanish state. By guiding the development of and eventual Olympic projection of national identity, Catalan and Spanish politicians hoped to create a resounding rallying point around which they could unite disparate individuals. The text is made up of: an introduction, five paragraphs on the different political perspective, conclusions, a commentary on the tallying of score and a note section with bibliographical references.