Purification and Preliminary Characterization of the TM0727 Protein from Thermotoga maritima

The TM0727 gene of Thermotoga maritima is responsible for encoding what has been reported to be a modulator of DNA gyrase (pmbA). Although the function of pmbA is still unknown, it is believedto be involved in cell division, carbon storage regulation, and the synthesis of the antibiotic peptide microcin B17. It is suggested that it serves together with tldD, a known zinc dependent protease, tomodulate DNA gyrase. TM0727 is believed to be a zinc dependent protease that binds zinc in the central active site of the molecule, located between two equivalent monomeric units. However, thecrystal structure determined by Wilson et al. (2005) did not contain zinc. It therefore remains to be seen if TM0727 requires zinc for activity, or regulation, and if the protein is indeed a protease. To begin studying this protein, the gene was expressed in BL21(DE3) pLysS cells and the induction time was optimized. Using affinity and ion exchange chromatography, the protein has been successfully purified. The purification procedure can be replicated to obtain sufficient protein for characterization. Purification results show that the protein loses stability after 24 hours and remains stable under an imidazole-free lysis workup. Preliminary characterization of TM0727 has focused on understanding the protein’s structuralproperties through tryptophan fluorescence anisotropy measurements. The four tryptophan residues located within the TM0727 dimer fluoresce at different maximum wavelengths and with differentintensities upon excitation with 295nm light. These emission properties are highly sensitive to the environment (solvent, surrounding residues) of each tryptophan residue. The low number oftryptophans allows for a specific monitoring of the protein’s structure as it denatures. As more denaturant is added to the protein, its tryptophan environments have clearly altered. This is indicative of unfolding and increased solvent exposure of the protein. This unfolding has been confirmed with the addition of a fluorescent quencher. Additionally, fluorescence anisotropy measurements have been carried out on the protein to gain a preliminary understanding of the rotational dynamics of the tryptophan residues. These experiments excite the tryptophan residues within the sample using a polarized light source. Polarized emission is then detected, the degree of which depends on the rotational dynamics and local environment of the tryptophan residues. The protein was denatured and the changes in emission were recorded to detect these structural changes. Results have shown a large change in quaternary structure, consistent with a dimer to monomer transition, occurs at 1.5M Guandidine HCl. There has also been an examination of the crystal structure for the location of a potential active site. The inner cavity of the protein was inspected visually to locate a potential location for a catalytic triad, specifically the amino acids found in the active sites of serine, cyteine, and aspartateproteases. It was found that a potential aspartic protease active site may be located between the Asparate286 and Aspartate287 residues. Further investigation is warranted to test this remotepossibility.

Immediate and delayed transplantation of mesenchymal stem cells improve muscle force after skeletal muscle injury in rats

Mesenchymal stem cell (MSC) therapy is a promising approach for regaining muscle function after trauma. Prior to clinical application, the ideal time of transplantation has to be determined. We investigated the effects of immediate and delayed transplantation. Sprague-Dawley rats received a crush trauma to the left soleus muscle. Treatment groups were transplanted locally with 2 × 10(6) autologous MSCs, either immediately or 7 days after trauma. Saline was used as sham therapy. Contraction force tests and histological analyses were performed 4 weeks after injury. GFP-labelled MSCs were followed after transplantation. The traumatized soleus muscles of the sham group displayed a reduction of twitch forces to 36 ± 17% and of tetanic forces to 29 ± 11% of the non-injured right control side, respectively. Delayed MSC transplantation resulted in a significant improvement of contraction maxima in both stimulation modes (twitch, p = 0.011; tetany, p = 0.014). Immediate transplantation showed a significant increase in twitch forces to 59 ± 17% (p = 0.043). There was no significant difference in contraction forces between muscles treated by immediate and delayed cell transplantation. We were able to identify MSCs in the interstitium of the injured muscles up to 4 weeks after transplantation. Despite the fundamental differences of the local environment, which MSCs encounter after transplantation, similar results could be obtained with respect to functional muscle regeneration. We believe that transplanted MSCs residing in the interstitial compartment evolve their regenerative capabilities through paracrine pathways. Our data suggest a large time window of the therapeutical measures.

Differential regulation of glucocorticoid synthesis in murine intestinal epithelial versus adrenocortical cell lines

Glucocorticoids are steroid hormones with important functions in development, immune regulation, and glucose metabolism. The adrenal glands are the predominant source of glucocorticoids; however, there is increasing evidence for extraadrenal glucocorticoid synthesis in thymus, brain, skin, and vascular endothelium. We recently identified intestinal epithelial cells as an important source of glucocorticoids, which regulate the activation of local intestinal immune cells. The molecular regulation of intestinal glucocorticoid synthesis is currently unexplored. In this study we investigated the transcriptional regulation of the steroidogenic enzymes P450 side-chain cleavage enzyme and 11beta-hydroxylase, and the production of corticosterone in the murine intestinal epithelial cell line mICcl2 and compared it with that in the adrenocortical cell line Y1. Surprisingly, we observed a reciprocal stimulation pattern in these two cell lines. Elevation of intracellular cAMP induced the expression of steroidogenic enzymes in Y1 cells, whereas it inhibited steroidogenesis in mICcl2 cells. In contrast, phorbol ester induced steroidogenic enzymes in intestinal epithelial cells, which was synergistically enhanced upon transfection of cells with the nuclear receptors steroidogenic factor-1 (NR5A1) and liver receptor homolog-1 (NR5A2). Finally, we observed that basal and liver receptor homolog-1/phorbol ester-induced expression of steroidogenic enzymes in mICcl2 cells was inhibited by the antagonistic nuclear receptor small heterodimer partner. We conclude that the molecular basis of glucocorticoid synthesis in intestinal epithelial cells is distinct from that in adrenal cells, most likely representing an adaptation to the local environment and different requirements.

Identifying indicators of sustainable development using the global sustainability quadrant approach

Advances in information technology and global data availability have opened the door for assessments of sustainable development at a truly macro scale. It is now fairly easy to conduct a study of sustainability using the entire planet as the unit of analysis; this is precisely what this work set out to accomplish. The study began by examining some of the best known composite indicator frameworks developed to measure sustainability at the country level today. Most of these were found to value human development factors and a clean local environment, but to gravely overlook consumption of (remote) resources in relation to nature’s capacity to renew them, a basic requirement for a sustainable state. Thus, a new measuring standard is proposed, based on the Global Sustainability Quadrant approach. In a two‐dimensional plot of nations’ Human Development Index (HDI) vs. their Ecological Footprint (EF) per capita, the Sustainability Quadrant is defined by the area where both dimensions satisfy the minimum conditions of sustainable development: an HDI score above 0.8 (considered ‘high’ human development), and an EF below the fair Earth‐share of 2.063 global hectares per person. After developing methods to identify those countries that are closest to the Quadrant in the present‐day and, most importantly, those that are moving towards it over time, the study tackled the question: what indicators of performance set these countries apart? To answer this, an analysis of raw data, covering a wide array of environmental, social, economic, and governance performance metrics, was undertaken. The analysis used country rank lists for each individual metric and compared them, using the Pearson Product Moment Correlation function, to the rank lists generated by the proximity/movement relative to the Quadrant measuring methods. The analysis yielded a list of metrics which are, with a high degree of statistical significance, associated with proximity to – and movement towards – the Quadrant; most notably: Favorable for sustainable development: use of contraception, high life expectancy, high literacy rate, and urbanization. Unfavorable for sustainable development: high GDP per capita, high language diversity, high energy consumption, and high meat consumption. A momentary gain, but a burden in the long‐run: high carbon footprint and debt. These results could serve as a solid stepping stone for the development of more reliable composite index frameworks for assessing countries’ sustainability.

Seasonal controls on grassland microbial biogeography: Are they governed by plants, abiotic properties or both?

Temporal dynamics create unique and often ephemeral conditions that can influence soil microbial biogeography at different spatial scales. This study investigated the relation between decimeter to meter spatial variability of soil microbial community structure, plant diversity, and soil properties at six dates from April through November. We also explored the robustness of these interactions over time. An historically unfertilized, unplowed grassland in southwest Germany was selected to characterize how seasonal variability in the composition of plant communities and substrate quality changed the biogeography of soil microorganisms at the plot scale (10 m x 10 m). Microbial community spatial structure was positively correlated with the local environment, i.e. physical and chemical soil properties, in spring and autumn, while the density and diversity of plants had an additional effect in the summer period. Spatial relationships among plant and microbial communities were detected only in the early summer and autumn periods when aboveground biomass increase was most rapid and its influence on soil microbial communities was greatest due to increased demand by plants for nutrients. Individual properties exhibited varying degrees of spatial structure over the season. Differential responses of Gram positive and Gram negative bacterial communities to seasonal shifts in soil nutrients were detected. We concluded that spatial distribution patterns of soil microorganisms change over a season and that chemical soil properties are more important controlling factors than plant density and diversity. Finer spatial resolution, such as the mm to cm scale, as well as taxonomic resolution of microbial groups, could help determine the importance of plant species density, composition, and growth stage in shaping microbial community composition and spatial patterns. (C) 2014 The Authors. Published by Elsevier Ltd. All rights reserved.

Comparison of two protocols of periosteal distraction osteogenesis in a rabbit calvaria model

The regenerative pathways during periosteal distraction osteogenesis may be influenced by the local environment composed by cells, growth factors, nutrition and mechanical load. The aim of the present study was to evaluate the influence of two protocols of periosteal distraction on bone formation. Custom made distraction devices were surgically fixed onto the calvariae of 60 rabbits. After an initial healing period of 7 days, two groups of animals were submitted to distraction rates of 0.25 and 0.5 mm/24 h for 10 days, respectively. Six animals per group were sacrificed 10 (mid-distraction), 17 (end-distraction), 24 (1-week consolidation), 31 (2-week consolidation) and 77 days (2-month consolidation) after surgery. Newly formed bone was assessed by means of micro-CT and histologically. Expression of transcripts encoding tissue-specific genes (BMP-2, RUNX2, ACP5, SPARC, collagen I α1, collagen II α1 and SOX9) was analyzed by quantitative PCR. Two patterns of bone formation were observed, originating from the old bone surface in Group I and from the periosteum in Group II. Bone volume (BV) and bone mineral density (BMD) significantly increased up to the 2-month consolidation period within the groups (p < 0.05). Significantly more bone was observed in Group II compared to Group I at the 2-month consolidation period (p < 0.001). Expression of transcripts encoding osteogenic genes in bone depended on the time-point of observation (p < 0.05). Low level of transcripts reveals an indirect role of periosteum in the osteogenic process. Two protocols of periosteal distraction in the present model resulted in moderate differences in terms of bone formation.

Pollen records of the Bykovsky Peninsula

New pollen and radiocarbon data from the Bykovsky Peninsula document the Late Pleistocene and Holocene environmental history of the Laptev Sea coast. More than 60 AMS-14C and conventional 14C dates indicate that the deposits accumulated during the last 60,000 radiocarbon yr BP. High concentration of green alga colonies (Pediustrum and Botryococcus) in the investigated sediment show that sedimentation was mostly in shallow water environments. Scarce grass and sedge communities dominated the vegetation 53-60 kyr BP. Climate was cold and dry. Open Poaceae and Cypcraccae associations with Asteraceae, Ranunculaceae, and Cichoriaceac, dominated in the area about 48-42.5 kyr BP. Steppic communities with Artemisia and shrubby tundra communities with Salix and Betula sect. Nanae were also present. Climate was dry, but relatively warm. Vegetation cover became denser about 42.5-33.5 kyr BP, reflecting more favorable climate conditions. Scarce Poaceae communities with some Caryophyllaceae, Asteraceae, Cichoriaceae, and Selaginella rupestris covered the Bykovsky Peninsula area during the Sartan (Late Weichselian) stage about 26-16 kyr BP. Disturbed, uncovered soils were very common in the area. Climate was extremely cold and dry. Poaceae and Cyperaceae associations with Caryophyllaceae, Asteraceae, Cichoriaceae dominated the vegetation in the late Sartan, ca 16-12.2 kyr BP. Climate was significantly warmer than in the early Sartan time. The lee Complex sedimentation was interrupted about 12 kyr BP; most likely it was connected with the beginning of the Allerod warnring. Shrubby (Betula sect. Nanae, Alnusfnuicosa, Salix, Ericales) tundra was widely distributed on the Bykovsky Peninsula during the early-middle Holacene. Climate was most favorable between 8200 and 4500 yr BP. Vegetation became similar to modern after 4500 yr BP, suggesting a deterioration of climate.

Wal-Mart, caballo de Troya de la globalización

Creada en 1962 por Sam Walton, una pequeña empresa de distribución norteamericana se convertirá rápidamente en la primera empresa mundial. Desde Bentonville, Arkansas, hasta el interior de China, Wal-Mart muestra un camino bastante clásico que va desde la sede principal de la empresa hacia una proliferación de sus tiendas en una quincena de países. Con una facturación de 405 mil millones de dólares en 2008, la empresa de distribución número uno en el mundo fue sobrepasada en abril de 2009 por la empresa petrolera Exxon Mobil en la clasificación de las quinientas empresas mundiales más importantes. Pero a diferencia de otras grandes transnacionales como Exxon Mobil o Microsoft, Wal-Mart, la primera empresa mundial de distribución, tiene que adaptarse al medio ambiente local con el fin de atraer a un máximo de consumidores. El artículo propone una aproximación a escala múltiple de las estrategias de desarrollo de esta firma internacional. Así, la aplicación de un análisis espacial es apropiada para esclarecer los vínculos existentes entre los territorios y las estrategias de los actores a diferentes escalas geográficas.

Wal-Mart, caballo de Troya de la globalización

Creada en 1962 por Sam Walton, una pequeña empresa de distribución norteamericana se convertirá rápidamente en la primera empresa mundial. Desde Bentonville, Arkansas, hasta el interior de China, Wal-Mart muestra un camino bastante clásico que va desde la sede principal de la empresa hacia una proliferación de sus tiendas en una quincena de países. Con una facturación de 405 mil millones de dólares en 2008, la empresa de distribución número uno en el mundo fue sobrepasada en abril de 2009 por la empresa petrolera Exxon Mobil en la clasificación de las quinientas empresas mundiales más importantes. Pero a diferencia de otras grandes transnacionales como Exxon Mobil o Microsoft, Wal-Mart, la primera empresa mundial de distribución, tiene que adaptarse al medio ambiente local con el fin de atraer a un máximo de consumidores. El artículo propone una aproximación a escala múltiple de las estrategias de desarrollo de esta firma internacional. Así, la aplicación de un análisis espacial es apropiada para esclarecer los vínculos existentes entre los territorios y las estrategias de los actores a diferentes escalas geográficas.

Wal-Mart, caballo de Troya de la globalización

Creada en 1962 por Sam Walton, una pequeña empresa de distribución norteamericana se convertirá rápidamente en la primera empresa mundial. Desde Bentonville, Arkansas, hasta el interior de China, Wal-Mart muestra un camino bastante clásico que va desde la sede principal de la empresa hacia una proliferación de sus tiendas en una quincena de países. Con una facturación de 405 mil millones de dólares en 2008, la empresa de distribución número uno en el mundo fue sobrepasada en abril de 2009 por la empresa petrolera Exxon Mobil en la clasificación de las quinientas empresas mundiales más importantes. Pero a diferencia de otras grandes transnacionales como Exxon Mobil o Microsoft, Wal-Mart, la primera empresa mundial de distribución, tiene que adaptarse al medio ambiente local con el fin de atraer a un máximo de consumidores. El artículo propone una aproximación a escala múltiple de las estrategias de desarrollo de esta firma internacional. Así, la aplicación de un análisis espacial es apropiada para esclarecer los vínculos existentes entre los territorios y las estrategias de los actores a diferentes escalas geográficas.

Alkenones and TEX86 temperature proxies for core-top sediments from the southern Adriatic Sea

The Mediterranean Sea is at the transition between temperate and tropical air masses and as such of importance for studying climate change. The Gulf of Taranto and adjacent SW Adriatic Sea are at the heart of this region. Their sediments are excellently suited for generating high quality environmental records for the last millennia with a sub-decadal resolution. The quality of these records is dependent on a careful calibration of the transfer functions used to translate the sedimentary lipid signals to the local environment. Here, we examine and calibrate the UK'37 and TEX86 lipid-based temperature proxies in 48 surface sediments and relate these to ambient sea surface temperatures and other environmental data. The UK'37-based temperatures in surface sediments reflect winter/spring sea surface temperatures in agreement with other studies demonstrating maximum haptophyte production during the colder season. The TEX86-based temperatures for the nearshore sites also reflect winter sea surface temperatures. However, at the most offshore sites, they correspond to summer sea surface temperatures. Additional lipid and environmental data including the distribution of the BIT index and remote-sensed chlorophyll-a suggest a shoreward increase of the impact of seasonal and spatial variability in nutrients and control of planktonic archaeal abundance by primary productivity, particle loading in surface waters and/or overprint by a cold-biased terrestrial TEX86 signal. As such the offshore TEX86 values seem to reflect a true summer signal to the effect that offshore UK'37 and TEX86 reconstruct winter and summer temperature, respectively, and hence provide information on the annual temperature amplitude.

Experimental and numerical analysis of reinforced concrete elements subjected to blast loading

El hormigón es uno de los materiales de construcción más empleados en la actualidad debido a sus buenas prestaciones mecánicas, moldeabilidad y economía de obtención, entre otras ventajas. Es bien sabido que tiene una buena resistencia a compresión y una baja resistencia a tracción, por lo que se arma con barras de acero para formar el hormigón armado, material que se ha convertido por méritos propios en la solución constructiva más importante de nuestra época. A pesar de ser un material profusamente utilizado, hay aspectos del comportamiento del hormigón que todavía no son completamente conocidos, como es el caso de su respuesta ante los efectos de una explosión. Este es un campo de especial relevancia, debido a que los eventos, tanto intencionados como accidentales, en los que una estructura se ve sometida a una explosión son, por desgracia, relativamente frecuentes. La solicitación de una estructura ante una explosión se produce por el impacto sobre la misma de la onda de presión generada en la detonación. La aplicación de esta carga sobre la estructura es muy rápida y de muy corta duración. Este tipo de acciones se denominan cargas impulsivas, y pueden ser hasta cuatro órdenes de magnitud más rápidas que las cargas dinámicas impuestas por un terremoto. En consecuencia, no es de extrañar que sus efectos sobre las estructuras y sus materiales sean muy distintos que las que producen las cargas habitualmente consideradas en ingeniería. En la presente tesis doctoral se profundiza en el conocimiento del comportamiento material del hormigón sometido a explosiones. Para ello, es crucial contar con resultados experimentales de estructuras de hormigón sometidas a explosiones. Este tipo de resultados es difícil de encontrar en la literatura científica, ya que estos ensayos han sido tradicionalmente llevados a cabo en el ámbito militar y los resultados obtenidos no son de dominio público. Por otra parte, en las campañas experimentales con explosiones llevadas a cabo por instituciones civiles el elevado coste de acceso a explosivos y a campos de prueba adecuados no permite la realización de ensayos con un elevado número de muestras. Por este motivo, la dispersión experimental no es habitualmente controlada. Sin embargo, en elementos de hormigón armado sometidos a explosiones, la dispersión experimental es muy acusada, en primer lugar, por la propia heterogeneidad del hormigón, y en segundo, por la dificultad inherente a la realización de ensayos con explosiones, por motivos tales como dificultades en las condiciones de contorno, variabilidad del explosivo, o incluso cambios en las condiciones atmosféricas. Para paliar estos inconvenientes, en esta tesis doctoral se ha diseñado un novedoso dispositivo que permite ensayar hasta cuatro losas de hormigón bajo la misma detonación, lo que además de proporcionar un número de muestras estadísticamente representativo, supone un importante ahorro de costes. Con este dispositivo se han ensayado 28 losas de hormigón, tanto armadas como en masa, de dos dosificaciones distintas. Pero además de contar con datos experimentales, también es importante disponer de herramientas de cálculo para el análisis y diseño de estructuras sometidas a explosiones. Aunque existen diversos métodos analíticos, hoy por hoy las técnicas de simulación numérica suponen la alternativa más avanzada y versátil para el cálculo de elementos estructurales sometidos a cargas impulsivas. Sin embargo, para obtener resultados fiables es crucial contar con modelos constitutivos de material que tengan en cuenta los parámetros que gobiernan el comportamiento para el caso de carga en estudio. En este sentido, cabe destacar que la mayoría de los modelos constitutivos desarrollados para el hormigón a altas velocidades de deformación proceden del ámbito balístico, donde dominan las grandes tensiones de compresión en el entorno local de la zona afectada por el impacto. En el caso de los elementos de hormigón sometidos a explosiones, las tensiones de compresión son mucho más moderadas, siendo las tensiones de tracción generalmente las causantes de la rotura del material. En esta tesis doctoral se analiza la validez de algunos de los modelos disponibles, confirmando que los parámetros que gobiernan el fallo de las losas de hormigón armado ante explosiones son la resistencia a tracción y su ablandamiento tras rotura. En base a los resultados anteriores se ha desarrollado un modelo constitutivo para el hormigón ante altas velocidades de deformación, que sólo tiene en cuenta la rotura por tracción. Este modelo parte del de fisura cohesiva embebida con discontinuidad fuerte, desarrollado por Planas y Sancho, que ha demostrado su capacidad en la predicción de la rotura a tracción de elementos de hormigón en masa. El modelo ha sido modificado para su implementación en el programa comercial de integración explícita LS-DYNA, utilizando elementos finitos hexaédricos e incorporando la dependencia de la velocidad de deformación para permitir su utilización en el ámbito dinámico. El modelo es estrictamente local y no requiere de remallado ni conocer previamente la trayectoria de la fisura. Este modelo constitutivo ha sido utilizado para simular dos campañas experimentales, probando la hipótesis de que el fallo de elementos de hormigón ante explosiones está gobernado por el comportamiento a tracción, siendo de especial relevancia el ablandamiento del hormigón. Concrete is nowadays one of the most widely used building materials because of its good mechanical properties, moldability and production economy, among other advantages. As it is known, it has high compressive and low tensile strengths and for this reason it is reinforced with steel bars to form reinforced concrete, a material that has become the most important constructive solution of our time. Despite being such a widely used material, there are some aspects of concrete performance that are not yet fully understood, as it is the case of its response to the effects of an explosion. This is a topic of particular relevance because the events, both intentional and accidental, in which a structure is subjected to an explosion are, unfortunately, relatively common. The loading of a structure due to an explosive event occurs due to the impact of the pressure shock wave generated in the detonation. The application of this load on the structure is very fast and of very short duration. Such actions are called impulsive loads, and can be up to four orders of magnitude faster than the dynamic loads imposed by an earthquake. Consequently, it is not surprising that their effects on structures and materials are very different than those that cause the loads usually considered in engineering. This thesis broadens the knowledge about the material behavior of concrete subjected to explosions. To that end, it is crucial to have experimental results of concrete structures subjected to explosions. These types of results are difficult to find in the scientific literature, as these tests have traditionally been carried out by armies of different countries and the results obtained are classified. Moreover, in experimental campaigns with explosives conducted by civil institutions the high cost of accessing explosives and the lack of proper test fields does not allow for the testing of a large number of samples. For this reason, the experimental scatter is usually not controlled. However, in reinforced concrete elements subjected to explosions the experimental dispersion is very pronounced. First, due to the heterogeneity of concrete, and secondly, because of the difficulty inherent to testing with explosions, for reasons such as difficulties in the boundary conditions, variability of the explosive, or even atmospheric changes. To overcome these drawbacks, in this thesis we have designed a novel device that allows for testing up to four concrete slabs under the same detonation, which apart from providing a statistically representative number of samples, represents a significant saving in costs. A number of 28 slabs were tested using this device. The slabs were both reinforced and plain concrete, and two different concrete mixes were used. Besides having experimental data, it is also important to have computational tools for the analysis and design of structures subjected to explosions. Despite the existence of several analytical methods, numerical simulation techniques nowadays represent the most advanced and versatile alternative for the assessment of structural elements subjected to impulsive loading. However, to obtain reliable results it is crucial to have material constitutive models that take into account the parameters that govern the behavior for the load case under study. In this regard it is noteworthy that most of the developed constitutive models for concrete at high strain rates arise from the ballistic field, dominated by large compressive stresses in the local environment of the area affected by the impact. In the case of concrete elements subjected to an explosion, the compressive stresses are much more moderate, while tensile stresses usually cause material failure. This thesis discusses the validity of some of the available models, confirming that the parameters governing the failure of reinforced concrete slabs subjected to blast are the tensile strength and softening behaviour after failure. Based on these results we have developed a constitutive model for concrete at high strain rates, which only takes into account the ultimate tensile strength. This model is based on the embedded Cohesive Crack Model with Strong Discontinuity Approach developed by Planas and Sancho, which has proved its ability in predicting the tensile fracture of plain concrete elements. The model has been modified for its implementation in the commercial explicit integration program LS-DYNA, using hexahedral finite elements and incorporating the dependence of the strain rate, to allow for its use in dynamic domain. The model is strictly local and does not require remeshing nor prior knowledge of the crack path. This constitutive model has been used to simulate two experimental campaigns, confirming the hypothesis that the failure of concrete elements subjected to explosions is governed by their tensile response, being of particular relevance the softening behavior of concrete.

De la desintegración Espacial a la Conformación regional en la Sabana de Bogotá. Una Propuesta para el desarrollo de la Ciudad Región en el siglo XXI

El presente trabajo de investigación aborda el tema del desarrollo regional en torno a una gran metrópoli como Bogotá y la Sabana de Bogotá. El crecimiento, expansión y relación con los municipios vecinos. Su entorno territorial; es un tema de discusión que cada día adquiere más fuerza desde hace varias décadas. Bogotá y la Sabana de Bogotá, se consideran en la actualidad como un caso representativo del caótico modelo de expansión urbana y crecimiento demográfico, enfrentado al desarrollo urbano como paradigma de la desigualdad de la ciudad latinoamericana. Son muchos los procesos y conflictos de naturaleza Regional y metropolitana que atraviesa la capital colombiana. Sin embargo esta investigación abordada el tema desde la necesidad de gobernanza y coordinación para el desarrollo territorial consensuado y equilibrado de la Región. La sabana de Bogotá está conformada por ciudades dormitorio, ciudades industriales, turísticas, agropecuarias, etc., es atravesada por el Río Bogotá, y en su centro la gran metrópoli, Bogotá D.C. centro de trabajo muy importante a nivel nacional, su área de influencia más próxima llega hasta: Caqueza, Zipaquira, Facatativa, Soacha, Fusagasuga y Girardot. Principales polos de desarrollo en la sabana y el departamento. Cundinamarca está compuesto por 15 provincias y Bogotá. Conformando un sistema de redes urbanas con necesidades comunes y demanda servicios públicos, de transporte, aseo, movilidad, salud, educación, espacio público y calidad ambiental. La metodología de esta investigación consiste en el análisis de la región a partir de la articulación de planes de ordenamiento territorial en un área de estudio propuesta. Mediante entrevistas con profesionales, expertos, funcionarios y empleados públicos y teniendo en cuenta las posibilidades legales en Colombia para el desarrollo territorial regional, establecer la situación real en materia de desarrollo para el departamento de Cundinamarca, evidenciando las necesidades del territorio y su desarrollo de una forma más compleja, valorando las sinergias y necesidades sociales, ambientales y económicas propias del crecimiento urbano, para proponer una serie de directrices que estructuren un desarrollo regional equilibrado en Bogotá y Cundinamarca. El análisis de los modelos del caso contribuye a fortalecer iniciativas para el desarrollo Regional de la Sabana de Bogotá como territorio sostenible: ambiental, económico y socialmente. En un sistema de redes que interconecte a Bogotá, con Cundinamarca, Colombia y el resto del mundo. Cundinamarca como región debe fijar estrategias y articular políticas en función de un modelo de desarrollo urbano regional para el departamento y la Sabana de Bogotá. Directrices departamentales básicas y fundamentales para el desarrollo territorial equilibrado que promueva ciudades sostenibles, compactas y con Calidad de vida para todos sus habitantes. ABSTRACT: This research addresses the issue of regional development around a big metropolis like Bogotá and Sabana de Bogota. The growth, expansion and relations with neighboring municipalities. Your local environment; It is an issue that becomes stronger every day for decades. Bogotá and Sabana de Bogotá, are considered today as a representative case of the chaotic model of urban expansion and population growth, urban development faced as a paradigm of inequality in Latin American city. Many processes and conflicts of Regional and metropolitan nature that crosses the Colombian capital. However this research addressed the issue from the need for governance and coordination for consensual and balanced territorial development of the region. The savannah of Bogota consists of bedroom communities, industrial cities, tourism, agriculture, etc., is crossed by the Bogota River, and at its center the great metropolis, Bogota DC center very important work at the national level, the area closest influence reaches: Caqueza, Zipaquira, Facatativa, Soacha, Fusagasuga and Girardot. Main centers of development in the savannah and the department. Cundinamarca is composed of 15 provinces and Bogota. Forming a system of urban networks with common needs and demand utilities, transportation, grooming, mobility, health, education, public space and environmental quality. The methodology of this research is the analysis of the region from the joint land use plans in the proposed study area. Through interviews with professionals, experts, public officials and employees and taking into account the legal possibilities in Colombia for regional territorial development, establish the real situation in development for the department of Cundinamarca, showing the region's needs and development of a more complex form, assessing synergies and own social, environmental and economic needs of urban growth, to propose a set of guidelines to structure a balanced regional development in Bogota and Cundinamarca. The analysis of case models helps to strengthen initiatives for regional development of the Sabana de Bogota and sustainable region: environmentally, economically and socially. In a network system that interconnects to Bogotá with Cundinamarca, Colombia and elsewhere. Cundinamarca region should set as joint strategies and policies based on a model of regional urban development for the department and the Sabana de Bogota. Basic and fundamental to balanced territorial development that fosters sustainable, compact and quality of life for all its inhabitants cities departmental guidelines.

HIP HOP COMO PROCESSO COMUNICACIONAL E SOCIABILIDADE PARA JOVENS INDÍGENAS DE DOURADOS – MS

Estudo sobre o Hip Hop como processo comunicacional e sociabilidade entre jovens indígenas de Dourados, Mato Grosso do Sul, para verificar quais os principais objetivos da prática do movimento Hip Hop, compreender se serve como comunicação, contribui para o fortalecimento da língua guarani ou gera novas tensões sociais na reserva. Para tanto, foi analisado aspectos históricos do movimento, passando pelos Estudos Culturais, e como Movimento Social, dando início à discussão de uma voz alternativa por meio do Hip Hop. Do ponto de vista metodológico, trata-se de um de estudo de caso, com representantes dos grupos de jovens Brô Mc's e Jovens Conscientes, das reservas Jaguapirú e Bororó, das etnias Guarani-Kaiowá de Dourados (MS). Foram realizadas entrevistas semiestruturadas junto a jovens que participaram das oficinas de hip hop, das lideranças indígenas e professores. A investigação é complementada pela pesquisa bibliográfica, documental e análises das letras de rap em confrontação com as visões da imprensa, a partir da análise dos jornais Diário MS e O Progresso. Os resultados apontam que os jovens se apropriam de uma cultura global para transformar o ambiente local com objetivo de preservar a língua guarani, uma alternativa para o conhecimento, logo para não seguirem caminhos como o das drogas. Negociando falas sobre sua realidade, dentro e fora da reserva, já que nos meios de comunicação locais há pouco espaço para a voz dos indígenas e dentro da reserva ainda há contestação do movimento em um contexto político, na tentativa de atingir uma cultura “pura”, devido à preocupação dos mais velhos com a perda de território.

Protein thermostability above 100°C: A key role for ionic interactions

The discovery of hyperthermophilic microorganisms and the analysis of hyperthermostable enzymes has established the fact that multisubunit enzymes can survive for prolonged periods at temperatures above 100°C. We have carried out homology-based modeling and direct structure comparison on the hexameric glutamate dehydrogenases from the hyperthermophiles Pyrococcus furiosus and Thermococcus litoralis whose optimal growth temperatures are 100°C and 88°C, respectively, to determine key stabilizing features. These enzymes, which are 87% homologous, differ 16-fold in thermal stability at 104°C. We observed that an intersubunit ion-pair network was substantially reduced in the less stable enzyme from T. litoralis, and two residues were then altered to restore these interactions. The single mutations both had adverse effects on the thermostability of the protein. However, with both mutations in place, we observed a fourfold improvement of stability at 104°C over the wild-type enzyme. The catalytic properties of the enzymes were unaffected by the mutations. These results suggest that extensive ion-pair networks may provide a general strategy for manipulating enzyme thermostability of multisubunit enzymes. However, this study emphasizes the importance of the exact local environment of a residue in determining its effects on stability.