Creating figures: why re-imagining urban structure supports a regenerative urban model

Some authors describe the contemporary metropolis like the field where co-exist simultaneous strategies of exploitation of fragmentary opportunities provided by the specific conditions of places (Florida, Bagnasco, Solá-Morales): physical (geography, infrastructures, etc.), social (people, culture and local values, etc.) and economic conditions. In their opinion, the causes of the ‘lateral’ development of the city could be (i) the relationships between the different elements of the land mosaic (Forman) and (ii) the fragmentary logics of the current urban realities. This important process of transformation would integrate the classic ‘lineal’ growth, more related to the urban rising along infrastructures. The result of these interactions is the change of scales in the performing of contemporary urban phenomena. Several authors have been interested in studying this new reality, called ‘exopolis’ (Soja), city-region (De las Rivas, Portas & al.) or metropolizated territory (Indovina, Monclús). Those not-conventional approaches are necessary to understand the contemporary urban condition and its complex, unstable, transient dynamics. Nowadays, several traditional concepts and ideas have become less useful and too rigid to achieve this target. This lack of effectiveness regards the discipline as a whole, divided between sectorial analysis and fragmentary solutions. In our opinion, using figures as ‘images with the potential to represent new territorial realities’ is one of the most important steps to produce an innovative and non-conventional understanding of post-metropolitan (Soja) urban space. This paper is aimed at explaining why figures are more useful than images to understand the complex urban pattern of current territory, as well as demonstrating this idea with the case studies of Valladolid and its emerging urban area. The result is a way to show the structure of this territory, which is more coherent with a contemporary narrative of space, and closer to its spatial and temporal dimensions. This is something not completely original but in this paper we present our views on it

The pURI family of expression vectors: A versatile set of ligation independent cloning plasmids for producing recombinant His-fusion proteins

A family of restriction enzyme- and ligation-independent cloning vectors has been developed for producing recombinant His-tagged fusion proteins in Escherichia coli. These are based on pURI2 and pURI3 expression vectors which have been previously used for the successful production of recombinant proteins at the milligram scale. The newly designed vectors combines two different promoters (lppp-5 and T7 RNA polymerase Ø10), two different endoprotease recognition sites for the His6-tag removal (enterokinase and tobacco etch virus), different antibiotic selectable markers (ampicillin and erythromycin resistance), and different placements of the His 6-tag (N- and C-terminus). A single gene can be cloned and further expressed in the eight pURI vectors by using six nucleotide primers, avoiding the restriction enzyme and ligation steps. A unique NotI site was introduced to facilitate the selection of the recombinant plasmid. As a case study, the new vectors have been used to clone the gene coding for the phenolic acid decarboxylase from Lactobacillus plantarum. Interestingly, the obtained results revealed markedly different production levels of the target protein, emphasizing the relevance of the cloning strategy on soluble protein production yield. Efficient purification and tag removal steps showed that the affinity tag and the protease cleavage sites functioned properly. The novel family of pURI vectors designed for parallel cloning is a useful and versatile tool for the production and purification of a protein of interest. © 2010 Elsevier Inc. All rights reserved.This work was supported by grants RM2008-00002 (Instituto Nacional de Investigación Agraria y Alimentaría), AGL2008-01052, Consolider INGENIO 2010 CSD2007-00063 FUN-C-FOOD (Comisión Interministerial de Ciencia y Tecnología), and S-0505/AGR/000153 and S2009/AGR-1469 (ALIBIRD) (Comunidad de Madrid). J.M.M. thanks the Ministerio de Ciencia e Innovación for a research grant (BFU2007-67404/BMC) and “Factoría de Cristalización” Consolider-Ingenio 2010 in support of his research. The technical assistance of M.V. Santamaría is greatly appreciated. J.A. Curiel is a recipient of a predoctoral fellowship from the MEC.Peer Reviewe

Ecos de Olmsted en Europa. El sistema de parques y los orígenes del urbanismo europeo contemporáneo

Con el diseño y la construcción del Emerald Necklace de Boston de Frederick Law Olmsted (1822-1903) tomaba forma una nueva herramienta urbanística directamente dirigida a adaptar el paisaje natural en el interior de la ciudad, concebida como sistema de parques, fusionando estructura urbana y diseño del paisaje. La idea sigue siendo esencial para pensar en una simbiosis entre ciudad y naturaleza. Sin ser exhaustivos, comprobamos como la idea se introduce en Europa de manera diversa y con logros dispares. A través de tres ‘paisajistas’ de generaciones que se suceden en el siglo XX, arraigados en contextos y lugares diferentes, queremos mostrar cómo el sistema de parques se consolida en Europa con perspectivas distintas aunque complementarias y que contribuyen a construir en la cultura urbano arquitectónica un camino de interacción entre ciudad y naturaleza, precedente directo de lo que hoy denominamos Green infrastructure.With the design and construction of the Emerald Necklace in Boston by Frederick Law Olmsted (1822-1903), a new urban tool took shape directly targeted at adapting natural landscapes to the interior of a city, conceptualized as a system of parks fused within an urban structure and landscape design. This idea continues to be key to the development process of creating symbiosis between city and nature. Without claiming to be exhaustive, this paper reviews how this idea was introduced to Europe in a variety of ways and through diverse achievements. Through three 20th century “landscapers” rooted in different contexts and places, this paper aims to show how systems of parks have consolidated in Europe with distinctive yet complimentary points-of-views, all contributing to the development of the interaction between city and nature in urban architectural culture, laying the foundation of what is presently called Green Infrastructure

The Lp_3561 and Lp_3562 enzymes support a functional divergence process in the lipase/esterase toolkit from Lactobacillus plantarum

Lactobacillus plantarum species is a good source of esterases since both lipolytic and esterase activities have been described for strains of this species. No fundamental biochemical difference exists among esterases and lipases since both share a common catalytic mechanism. L. plantarum WCFS1 possesses a protein, Lp_3561, which is 44% identical to a previously described lipase, Lp_3562. In contrast to Lp_3562, Lp_3561 was unable to degrade esters possessing a chain length higher than C4 and the triglyceride tributyrin. As in other L. plantarum esterases, the electrostatic potential surface around the active site in Lp_3561 is predicted to be basic, whereas it is essentially neutral in the Lp_3562 lipase. The fact that the genes encoding both proteins were located contiguously in the L. plantarum WCFS1 genome, suggests that they originated by tandem duplication, and therefore are paralogs as new functions have arisen during evolution. The presence of the contiguous lp_3561 and lp_3562 genes was studied among L. plantarum strains. They are located in a 8,903 bp DNA fragment that encodes proteins involved in the catabolism of sialic acid and are predicted to increase bacterial adaptability under certain growth conditions

Transcriptomic effects of Tet-on and mifepristone-inducible systems in mouse liver

Control of transgene expression from long-term expression vectors can be achieved with inducible and regulated promoters. The two most commonly used inducible systems employ doxycycline or mifepristone as the drug activating a silent trans-activator, which is expressed from a constitutive promoter. We evaluated the alterations provoked by constitutive expression in the liver of rtTA2(S)-M2 (rtTA2; second-generation reverse tetracycline-controlled trans-activator) and GLp65, which are the trans-activators of the doxycyline- and mifepristone-inducible systems, respectively. To this end we performed transcriptomic analysis of mice expressing these trans-activators in the liver over 1 month. rtTA2 expression induced alterations in a few genes (69 gene probesets; false discovery rate [FDR], approximately 0.05), whereas GLp65 caused more numerous changes (1059 gene probe-sets, an FDR of approximately 0.05). However, only 20 and 53 of the genes from the rtTA2 and GLp65 groups, respectively, showed changes (R-fold >or= 3). Functional assignments indicate that alterations were mild and of little general significance. Few additional transcriptomic changes were observed when expressing trans-activators in the presence of inducer drugs; most were due to the drugs themselves. These results and the absence of toxicity observed in treated animals indicate that the two inducible systems are well tolerated and have little impact on the liver transcriptome profile. The milder alterations found with the use of rtTA2 suggest that this system is possibly safer for gene therapy application

Transcriptomic effects of Tet-on and mifepristone-inducible systems in mouse liver

Control of transgene expression from long-term expression vectors can be achieved with inducible and regulated promoters. The two most commonly used inducible systems employ doxycycline or mifepristone as the drug activating a silent trans-activator, which is expressed from a constitutive promoter. We evaluated the alterations provoked by constitutive expression in the liver of rtTA2(S)-M2 (rtTA2; second-generation reverse tetracycline-controlled trans-activator) and GLp65, which are the trans-activators of the doxycyline- and mifepristone-inducible systems, respectively. To this end we performed transcriptomic analysis of mice expressing these trans-activators in the liver over 1 month. rtTA2 expression induced alterations in a few genes (69 gene probesets; false discovery rate [FDR], approximately 0.05), whereas GLp65 caused more numerous changes (1059 gene probe-sets, an FDR of approximately 0.05). However, only 20 and 53 of the genes from the rtTA2 and GLp65 groups, respectively, showed changes (R-fold >or= 3). Functional assignments indicate that alterations were mild and of little general significance. Few additional transcriptomic changes were observed when expressing trans-activators in the presence of inducer drugs; most were due to the drugs themselves. These results and the absence of toxicity observed in treated animals indicate that the two inducible systems are well tolerated and have little impact on the liver transcriptome profile. The milder alterations found with the use of rtTA2 suggest that this system is possibly safer for gene therapy application