Modeling impact assessment of transport policies through a multiregional input-output integrated approach

El objetivo de esta investigación es desarrollar una metodología para estimar los potenciales impactos económicos y de transporte generados por la aplicación de políticas en el sector transporte. Los departamentos de transporte y otras instituciones gubernamentales relacionadas se encuentran interesadas en estos análisis debido a que son presentados comúnmente de forma errónea por la insuficiencia de datos o por la falta de metodologías adecuadas. La presente investigación tiene por objeto llenar este vacío haciendo un análisis exhaustivo de las técnicas disponibles que coincidan con ese propósito. Se ha realizado un análisis que ha identificado las diferencias cuando son aplicados para la valoración de los beneficios para el usuario o para otros efectos como aspectos sociales. Como resultado de ello, esta investigación ofrece un enfoque integrado que incluye un modelo Input-Output de múltiples regiones basado en la utilidad aleatoria (RUBMRIO), y un modelo de red de transporte por carretera. Este modelo permite la reproducción con mayor detalle y realismo del transporte de mercancías que por medio de su estructura sectorial identifica los vínculos de las compras y ventas inter-industriales dentro de un país utilizando los servicios del transporte de mercancías. Por esta razón, el modelo integrado es aplicable a diversas políticas de transporte. En efecto, el enfoque se ha aplicado para estudiar los efectos macroeconómicos regionales de la implementación de dos políticas diferentes en el sistema de transporte de mercancías de España, tales como la tarificación basada en la distancia recorrida por vehículo-kilómetro (€/km) aplicada a los vehículos del transporte de mercancías, y para la introducción de vehículos más largos y pesados de mercancías en la red de carreteras de España. El enfoque metodológico se ha evaluado caso por caso teniendo en cuenta una selección de la red de carreteras que unen las capitales de las regiones españolas. También se ha tenido en cuenta una dimensión económica a través de una tabla Input-Output de múltiples regiones (MRIO) y la base de datos de conteo de tráfico existente para realizar la validación del modelo. El enfoque integrado reproduce las condiciones de comercio observadas entre las regiones usando el sistema de transporte de mercancías por carretera, y que permite por comparación con los escenarios de políticas, determinar las contribuciones a los cambios distributivos y generativos. Así pues, el análisis estima los impactos económicos en cualquier región considerando los cambios en el Producto Interno Bruto (PIB) y el empleo. El enfoque identifica los cambios en el sistema de transporte a través de todos los caminos de la red de transporte a través de las medidas de efectividad (MOEs). Los resultados presentados en esta investigación proporcionan evidencia sustancial de que en la evaluación de las políticas de transporte, es necesario establecer un vínculo entre la estructura económica de las regiones y de los servicios de transporte. Los análisis muestran que para la mayoría de las regiones del país, los cambios son evidentes para el PIB y el empleo, ya que el comercio se fomenta o se inhibe. El enfoque muestra cómo el tráfico se desvía en ambas políticas, y también determina detalles de las emisiones de contaminantes en los dos escenarios. Además, las políticas de fijación de precios o de regulación de los sistemas de transporte de mercancías por carretera dirigidas a los productores y consumidores en las regiones promoverán transformaciones regionales afectando todo el país, y esto conduce a conclusiones diferentes. Así mismo, este enfoque integrado podría ser útil para evaluar otras políticas y otros países en todo el mundo. The purpose of this research is to develop a methodological approach aimed at assessing the potential economic and transportation impacts of transport policies. Transportation departments and other related government parties are interested in such analysis because it is commonly misrepresented for the insufficiency of data and suitable methodologies available. This research is directed at filling this gap by making a comprehensive analysis of the available techniques that match with that purpose. The differences when they are applied for the valuation of user benefits or for other impacts as social matters have been identified. As a result, this research presents an integrated approach which includes both a random utility-based multiregional Input-Output model (RUBMRIO), and a road transport network model. This model accounts for freight transport with more detail and realism because its commodity-based structure traces the linkages of inter-industry purchases and sales that use freight services within a given country. For this reason, the integrated model is applicable to various transport policies. In fact, the approach is applied to study the regional macroeconomic effects of implementing two different policies in the freight transport system of Spain, such as a distance-based charge in vehicle-kilometer (€/km) for Heavy Goods Vehicles (HGVs), and the introduction of Longer and Heavier Vehicles (LHVs) in the road network of Spain. The methodological approach has been evaluated on a case by case basis considering a selected road network of highways linking the capitals of the Spanish regions. It has also considered an economic dimension through a Multiregional Input Output Table (MRIO) and the existing traffic count database used in the model validation. The integrated approach replicates observed conditions of trade among regions using road freight transport systems that determine contributions to distributional and generative changes by comparison with policy scenarios. Therefore, the model estimates economic impacts in any given area by considering changes in Gross Domestic Product (GDP), employment (jobs), and in the transportation system across all paths of the transport network considering Measures of effectiveness (MOEs). The results presented in this research provide substantive evidence that in the assessment of transport policies it is necessary to establish a link between the economic structure of regions and the transportation services. The analysis shows that for most regions in the country, GDP and employment changes are noticeable when trade is encouraged or discouraged. This approach shows how traffic is diverted in both policies, and also provides details of the pollutant emissions in both scenarios. Furthermore, policies, such as pricing or regulation of road freight transportation systems, directed to producers and consumers in regions will promote different regional transformations across the country, and this lead to different conclusions. In addition, this integrated approach could be useful to assess other policies and countries worldwide.

Modelización de los Procesos de Cristalización en Poliolefinas

“Por lo tanto, la cristalización de polímeros se supone, y en las teorías se describe a menudo, como un proceso de múltiples pasos con muchos aspectos físico-químicos y estructurales influyendo en él. Debido a la propia estructura de la cadena, es fácil entender que un proceso que es termodinámicamente forzado a aumentar su ordenamiento local, se vea obstaculizado geométricamente y, por tanto, no puede conducirse a un estado de equilibrio final. Como resultado, se forman habitualmente estructuras de no equilibrio con diferentes características dependiendo de la temperatura, presión, cizallamiento y otros parámetros físico-químicos del sistema”. Estas palabras, pronunciadas recientemente por el profesor Bernhard Wunderlich, uno de los mas relevantes fisico-quimicos que han abordado en las ultimas décadas el estudio del estado físico de las macromoléculas, adelantan lo que de alguna manera se explicita en esta memoria y constituyen el “leitmotiv” de este trabajo de tesis. El mecanismo de la cristalización de polímeros esta aun bajo debate en la comunidad de la física de polímeros y la mayoría de los abordajes experimentales se explican a través de la teoría LH. Esta teoría clásica debida a Lauritzen y Hoffman (LH), y que es una generalización de la teoría de cristalización de una molécula pequeña desde la fase de vapor, describe satisfactoriamente muchas observaciones experimentales aunque esta lejos de explicar el complejo fenómeno de la cristalización de polímeros. De hecho, la formulación original de esta teoría en el National Bureau of Standards, a comienzos de la década de los 70, sufrió varias reformulaciones importantes a lo largo de la década de los 80, buscando su adaptación a los hallazgos experimentales. Así nació el régimen III de cristalización que posibilita la creacion de nichos moleculares en la superficie y que dio pie al paradigma ofrecido por Sadler y col., para justificar los experimentos que se obtenian por “scattering” de neutrones y otras técnicas como la técnica de “droplets” o enfriamiento rapido. Por encima de todo, el gran éxito de la teoría radica en que explica la dependencia inversa entre el tamaño del plegado molecular y el subenfriamiento, definido este ultimo como el intervalo de temperatura que media entre la temperatura de equilibrio y la temperatura de cristalización. El problema concreto que aborda esta tesis es el estudio de los procesos de ordenamiento de poliolefinas con distinto grado de ramificacion mediante simulaciones numéricas. Los copolimeros estudiados en esta tesis se consideran materiales modelo de gran homogeneidad molecular desde el punto de vista de la distribución de tamaños y de ramificaciones en la cadena polimérica. Se eligieron estas poliolefinas debido al gran interes experimental en conocer el cambio en las propiedades fisicas de los materiales dependiendo del tipo y cantidad de comonomero utilizado. Además, son modelos sobre los que existen una ingente cantidad de información experimental, que es algo que preocupa siempre al crear una realidad virtual como es la simulación. La experiencia en el grupo Biophym es que los resultados de simulación deben de tener siempre un correlato mas o menos próximo experimental y ese argumento se maneja a lo largo de esta memoria. Empíricamente, se conoce muy bien que las propiedades físicas de las poliolefinas, en suma dependen del tipo y de la cantidad de ramificaciones que presenta el material polimérico. Sin embargo, tal como se ha explicado no existen modelos teóricos adecuados que expliquen los mecanismos subyacentes de los efectos de las ramas. La memoria de este trabajo es amplia por la complejidad del tema. Se inicia con una extensa introducción sobre los conceptos básicos de una macromolecula que son relevantes para entender el contenido del resto de la memoria. Se definen los conceptos de macromolecula flexible, distribuciones y momentos, y su comportamiento en disolución y fundido con los correspondientes parametros caracteristicos. Se pone especial énfasis en el concepto de “entanglement” o enmaranamiento por considerarse clave a la hora de tratar macromoléculas con una longitud superior a la longitud critica de enmaranamiento. Finaliza esta introducción con una reseña sobre el estado del arte en la simulación de los procesos de cristalización. En un segundo capitulo del trabajo se expone detalladamente la metodología usada en cada grupo de casos. En el primer capitulo de resultados, se discuten los estudios de simulación en disolución diluida para sistemas lineales y ramificados de cadena única. Este caso mas simple depende claramente del potencial de torsión elegido tal como se discute a lo largo del texto. La formación de los núcleos “babys” propuestos por Muthukumar parece que son consecuencia del potencial de torsión, ya que este facilita los estados de torsión mas estables. Así que se propone el análisis de otros potenciales que son igualmente utilizados y los resultados obtenidos sobre la cristalización, discutidos en consecuencia. Seguidamente, en un segundo capitulo de resultados se estudian moleculas de alcanos de cadena larga lineales y ramificados en un fundido por simulaciones atomisticas como un modelo de polietileno. Los resultados atomisticos pese a ser de gran detalle no logran captar en su totalidad los efectos experimentales que se observan en los fundidos subenfriados en su etapa previa al estado ordenado. Por esta razon se discuten en los capítulos 3 y 4 de resultados sistemas de cadenas cortas y largas utilizando dos modelos de grano grueso (CG-PVA y CG-PE). El modelo CG-PE se desarrollo durante la tesis. El uso de modelos de grano grueso garantiza una mayor eficiencia computacional con respecto a los modelos atomísticos y son suficientes para mostrar los fenómenos a la escala relevante para la cristalización. En todos estos estudios mencionados se sigue la evolución de los procesos de ordenamiento y de fusión en simulaciones de relajación isoterma y no isoterma. Como resultado de los modelos de simulación, se han evaluado distintas propiedades fisicas como la longitud de segmento ordenado, la cristalinidad, temperaturas de fusion/cristalizacion, etc., lo que permite una comparación con los resultados experimentales. Se demuestra claramente que los sistemas ramificados retrasan y dificultan el orden de la cadena polimérica y por tanto, las regiones cristalinas ordenadas decrecen al crecer las ramas. Como una conclusión general parece mostrarse una tendencia a la formación de estructuras localmente ordenadas que crecen como bloques para completar el espacio de cristalización que puede alcanzarse a una temperatura y a una escala de tiempo determinada. Finalmente hay que señalar que los efectos observados, estan en concordancia con otros resultados tanto teoricos/simulacion como experimentales discutidos a lo largo de esta memoria. Su resumen se muestra en un capitulo de conclusiones y líneas futuras de investigación que se abren como consecuencia de esta memoria. Hay que mencionar que el ritmo de investigación se ha acentuado notablemente en el ultimo ano de trabajo, en parte debido a las ventajas notables obtenidas por el uso de la metodología de grano grueso que pese a ser muy importante para esta memoria no repercute fácilmente en trabajos publicables. Todo ello justifica que gran parte de los resultados esten en fase de publicación. Abstract “Polymer crystallization is therefore assumed, and in theories often described, to be a multi step process with many influencing aspects. Because of the chain structure, it is easy to understand that a process which is thermodynamically forced to increase local ordering but is geometrically hindered cannot proceed into a final equilibrium state. As a result, nonequilibrium structures with different characteristics are usually formed, which depend on temperature, pressure, shearing and other parameters”. These words, recently written by Professor Bernhard Wunderlich, one of the most prominent researchers in polymer physics, put somehow in value the "leitmotiv "of this thesis. The crystallization mechanism of polymers is still under debate in the physics community and most of the experimental findings are still explained by invoking the LH theory. This classical theory, which was initially formulated by Lauritzen and Hoffman (LH), is indeed a generalization of the crystallization theory for small molecules from the vapor phase. Even though it describes satisfactorily many experimental observations, it is far from explaining the complex phenomenon of polymer crystallization. This theory was firstly devised in the early 70s at the National Bureau of Standards. It was successively reformulated along the 80s to fit the experimental findings. Thus, the crystallization regime III was introduced into the theory in order to explain the results found in neutron scattering, droplet or quenching experiments. This concept defines the roughness of the crystallization surface leading to the paradigm proposed by Sadler et al. The great success of this theory is the ability to explain the inverse dependence of the molecular folding size on the supercooling, the latter defined as the temperature interval between the equilibrium temperature and the crystallization temperature. The main scope of this thesis is the study of ordering processes in polyolefins with different degree of branching by using computer simulations. The copolymers studied along this work are considered materials of high molecular homogeneity, from the point of view of both size and branching distributions of the polymer chain. These polyolefins were selected due to the great interest to understand their structure– property relationships. It is important to note that there is a vast amount of experimental data concerning these materials, which is essential to create a virtual reality as is the simulation. The Biophym research group has a wide experience in the correlation between simulation data and experimental results, being this idea highly alive along this work. Empirically, it is well-known that the physical properties of the polyolefins depend on the type and amount of branches presented in the polymeric material. However, there are not suitable models to explain the underlying mechanisms associated to branching. This report is extensive due to the complexity of the topic under study. It begins with a general introduction to the basics concepts of macromolecular physics. This chapter is relevant to understand the content of the present document. Some concepts are defined along this section, among others the flexibility of macromolecules, size distributions and moments, and the behavior in solution and melt along with their corresponding characteristic parameters. Special emphasis is placed on the concept of "entanglement" which is a key item when dealing with macromolecules having a molecular size greater than the critical entanglement length. The introduction finishes with a review of the state of art on the simulation of crystallization processes. The second chapter of the thesis describes, in detail, the computational methodology used in each study. In the first results section, we discuss the simulation studies in dilute solution for linear and short chain branched single chain models. The simplest case is clearly dependent on the selected torsion potential as it is discussed throughout the text. For example, the formation of baby nuclei proposed by Mutukhumar seems to result from the effects of the torsion potential. Thus, we propose the analysis of other torsion potentials that are also used by other research groups. The results obtained on crystallization processes are accordingly discussed. Then, in a second results section, we study linear and branched long-chain alkane molecules in a melt by atomistic simulations as a polyethylene-like model. In spite of the great detail given by atomistic simulations, they are not able to fully capture the experimental facts observed in supercooled melts, in particular the pre-ordered states. For this reason, we discuss short and long chains systems using two coarse-grained models (CG-PVA and CG-PE) in section 3 and 4 of chapter 2. The CG-PE model was developed during the thesis. The use of coarse-grained models ensures greater computational efficiency with respect to atomistic models and is enough to show the relevant scale phenomena for crystallization. In all the analysis we follow the evolution of the ordering and melting processes by both isothermal and non isothermal simulations. During this thesis we have obtained different physical properties such as stem length, crystallinity, melting/crystallization temperatures, and so on. We show that branches in the chains cause a delay in the crystallization and hinder the ordering of the polymer chain. Therefore, crystalline regions decrease in size as branching increases. As a general conclusion, it seems that there is a tendency in the macromolecular systems to form ordered structures, which can grown locally as blocks, occupying the crystallization space at a given temperature and time scale. Finally it should be noted that the observed effects are consistent with both, other theoretical/simulation and experimental results. The summary is provided in the conclusions chapter along with future research lines that open as result of this report. It should be mentioned that the research work has speeded up markedly in the last year, in part because of the remarkable benefits obtained by the use of coarse-grained methodology that despite being very important for this thesis work, is not easily publishable by itself. All this justify that most of the results are still in the publication phase.

Bandas programáticas: Variaciones de una estructura organizativa en la obra de Rem Koolhaas/ OMA.

Resulta posible extraer una constante en los proyectos de Rem Koolhaas / OMA, un sistema de bandas, al que a menudo recurre, como estructura de organización programática. Su germen en "The Berlin Wall as Architecture", su conexión con Delirious New York, y la persistencia hasta los grandes propuestas desarrolladas en los últimos años como el LACMA (2001), la Biblioteca de Seattle(1999-04) o el CCTV(2002-8). El presente trabajo se desarrolla bajo una doble hipótesis: La primera es que este esquema de organización se pone en valor originalmente en la aproximación a “The Berlin Wall as Architecture" que Koolhaas realiza en el 1971 como estudiante en la A.A. Y la segunda, que este sistema de bandas programáticas, va transformándose, desde su concepción como esquema organizativo en la Spear House (1974), a diagrama operativo en proyectos como la Villette (1982), Melun- Sénart (1987), ZKM (1989), hasta un sistema de bandas con connotaciones más figurativas que estratégicas en la Biblioteca de Seattle, y al que Peter Eisenman se referirá como diagrama icónico. Para realizar este estudio, se opera con una selección de obras representativas, a partir de esta constante (sistema de bandas) para analizar en qué medida aparece y se transforma. EN It is possible to extract extract a constant in Rem Koolhaas / OMA´s projects, a system of bands, which often resorts such as programmatic organization structure. The germ in "The Berlin Wall as Architecture"(1971), his connection with Delirious New York(1972-78), and persistence to large proposals developed in recent years as the LACMA (2001), the Seattle Library (1999- 1904) or the CCTV (2002-8). This work was conducted under a double hypothesis: The first is that this organizational scheme is put in value approach originally in "The Berlin Wall as Architecture" Koolhaas made in 1971 as a student at the AA.Tthe second, which this band program system, is transformed, from its conception as organizational scheme in Spear House (1974), a chart operating in projects such as Villette (1982), Melun sénart (1987), ZKM (1989), to a system bands with figurative connotations strategic Seattle Library, and Peter Eisenman referred as iconic diagram. For this study, it operates with a selection of representative works from this constant (band system) to analyze to what extent appears and transforms.

The HOSHIN KANRI TREE. Cross-Plant Lean Shopfloor Management

Shopfloor Management (SM) empowerment methodologies have traditionally focused on two aspects: goal achievement following rigid structures, such as SQDCME, or evolutional aspects of empowerment factors away from strategic goal achievement. Furthermore, SM Methodologies have been organized almost solely around the hierarchical structure of the organization, failing systematically to cope with the challenges that Industry 4.0 is facing. The latter include the growing complexity of value-stream networks, sustainable empowerment of the workforce (Learning Factory), an autonomous and intelligent process management (Smart Factory), the need to cope with the increasing complexity of value-stream networks (VSN) and the leadership paradigm shift to strategic alignment. This paper presents a novel Lean SM Method (LSM) called ?HOSHIN KANRI Tree? (HKT), which is based on standardization of the communication patterns among process owners (POs) by PDCA. The standardization of communication patterns by HKT technology should bring enormous benefits in value stream (VS) performance, speed of standardization and learning rates to the Industry 4.0 generation of organizations. These potential advantages of HKT are being tested at present in worldwide research.

Aspectos formales y constructivos en la obra de Guillem Sagrera: el uso de las plantillas

Los enjarjes, jarjas o jarjamentos conforman el comienzo de la bóveda de crucería, el lugar en el que todos los nervios nacen reunidos. Permiten la transición entre el muro y las dovelas que constituyen nervios, resolviendo un encuentro de gran importancia constructiva. Están compuestos por piezas de lechos horizontales que traban con el muro y que, además de conformar el arranque de los nervios, han de proporcionar apoyo a las primeras dovelas. Al construirse a la vez que el muro y antes que el resto de la bóveda, se podría decir que son los encargados de portar su ADN: en ellos queda registrado el número de nervios que tendrá la bóveda, su curvatura, desde qué altura arrancará cada uno, su perfil, etc. El objetivo de este trabajo se centra en estudio de los enjarjes en la obra de Guillem Sagrera abordando su dimensión tecnológica, constructiva y de diseño. Dos características los distinguen de otras soluciones y justifican su interés: los nervios surgen directamente del muro, sin la intermediación de capiteles, ménsulas o pilastras; y se diseñan con la intención de facilitar la aparición de intersecciones entre las molduras de sus perfiles, en algunos casos mediante el cruce de los ejes de los nervios. Sagrera nos acerca a una innovación arquitectónica en la que el esfuerzo no se centra en realizar bóvedas con muchos nervios o con trazados en planta complejos, sino en la cuidada resolución de este encuentro de nervios. En ella se adivina el trabajo minucioso con las plantillas que controlan el trazado del contorno de los lechos de las piezas y la hábil mano de los canteros que son capaces de tallar intersecciones de gran complejidad. Se ha realizado un repaso de los primeros experimentos relativos al modo de relacionarse los nervios entre sí para ilustrar el contexto y origen de las soluciones realizadas por Guillem Sagrera. Mostramos que ante ciertas dificultades, consecuencia de la reunión de nervios, los constructores fueron capaces de desarrollar nuevas soluciones, mediante la experimentación con un sistema constructivo que conocían y manejaban con destreza. Para acercarnos a la comprensión de las estrategias de diseño que permiten el proyecto de estos enjarjes y los procedimientos técnicos y constructivos necesarios para su ejecución, nos vimos en la necesidad de adentrarnos en la problemática general de los enjarjes de la bóveda de crucería. De este modo, lo que empezó siendo una introducción para poder contextualizar la obra del mallorquín acabó convirtiéndose en la primera parte de la tesis, cuyo volumen prácticamente equipara a la segunda. En ella presentamos el proceso de diseño, trazado y talla de los cuatro enjarjes llevados a cabo en el taller de cantería de la ETSAM, en los que hemos podido experimentar de manera práctica los aspectos teóricos desarrollados. Estos ensayos nos han permitido contrastar hipótesis y baremar la dificultad de ciertos procedimientos o procesos, así como acercarnos realmente al elemento constructivo. El trabajo práctico nos ha enseñado a no fiarnos siempre de las hipótesis que se desarrollan modelando con el ordenador o dibujando; a valorar el pensar con las manos. En relación con la obra de Sagrera, la presente investigación realiza aportaciones al conocimiento del cambio proyectual y constructivo llevado a cabo en los arranques de las bóvedas entre los siglos XIII y XV, cuando los nervios comienzan a surgir directamente de los soportes y se dan los primeros cruzamientos. Mostramos que ya no solamente se construyen enjarjes fruto directo de la geometría general de la bóveda, sino que se llevan a cabo cambios deliberados en relación a su resultado, en los que se advierten decisiones proyectuales que, por supuesto, no serían viables sin las posibilidades que ofrece el trabajo con plantillas. ABSTRACT The solid blocks commonly known as tas-de-charge (in Spanish enjarjes, jarjas or jarjamentos) constitute the beginning of the ribbed vault – the place from which all the ribs spring together. They facilitate a transition between the wall and the rib voussoirs, and thus solve a junction of utmost constructive importance. They consist of blocks set in horizontal courses which interlock with the wall and which, as well as constituting the springing of the ribs, serve as a support for their first voussoirs. The tas-de-charge are built simultaneously with the wall and well before the remainder of the vault – thus, they arguably carry its ‘DNA’, since they register how many ribs the vault will have as well as their curvature, their springing height or their profile. This work is focused on the study of the tas-de-charge in the works of Guillem Sagrera, and will address their technological, constructive and design aspects. Two characteristics set these apart from other solutions and justify their relevance: these are that the ribs spring directly from the wall without the mediation of capitals, corbels or pilasters; and that they are deliberately designed to force the intersection of their mouldings, in some cases by crossing the rib axes. Sagrera’s work tells a story of architectural innovation – one where the effort is not centred on creating vaults with numerous ribs or a sophisticated ground plan, but on carefully solving the rib unions, which evidence a meticulous use of templates to control the tracing of the pieces’ profiles as well as the skill of the stonemasons, able to carve highly complex intersections. An overview of the first experiments with rib relationships will illustrate the context and origin of Guillem Sagrera’s solutions. We show how, faced with difficulties arising from the convergence of ribs into the tas-de-charge, builders were able to develop new solutions by experimenting with a construction system that they were already familiar with and could control easily. In order to gain a better insight into the design strategies behind his tas-de-charge and the technical and constructive procedures required for their execution, we found ourselves facing the need to address the general subject of tas-de-charge in ribbed vaults. This, which began as an introduction meant as context for Sagrera’s work, took on a life of its own and became the first half of the thesis, with a volume practically equal to that of the second. We have devoted a chapter to experimental archaeology. It comprises the design, tracing and carving processes for the four tas-de-charge executed at the ETSAM Stonecutting Workshop, in which we have tested experimentally the theories studied in the previous chapters. These tests have allowed us to contrast hypotheses, assess the difficulty of certain procedures or processes and understand the built element as a real entity. The practical work has taught us not to always trust the hypotheses proposed through computer modelling or drawing – and to recognise the importance of coordinating the hands and the mind. After studying Sagrera’s work and contrasting it with other related or previous tas-de-charge, our research will seek to make a contribution to the study of the shift in the design and construction of vault springers that took place between the 13th and 14th centuries, when ribs began to spring directly from their support and moulding crossings began to appear. We show that, from then on, tas-de-charge would not only depend on the general vault geometry – deliberate modifications would be carried out in order to achieve the desired result. This reveals design decisions that would have been unworkable if not for the effective use of template strategies.

On the reaction mechanism of l-lactate oxidase: Quantitative structure-activity analysis of the reaction with para-substituted l-mandelates

The rate constants for reduction of the flavoenzyme, l-lactate oxidase, and a mutant (in which alanine 95 is replaced by glycine), by a series of para-substituted mandelates, in both the 2-1H- and 2-2H- forms, have been measured by rapid reaction spectrophotometry. In all cases, significant isotope effects (1H/2H = 3–7) on the rate constants of flavin reduction were found, indicating that flavin reduction is a direct measure of α-C-H bond breakage. The rate constants show only a small influence of the electronic characteristics of the substituents, but show a good correlation when combined with some substituent volume parameters. A surprisingly good correlation is found with the molecular mass of the substrate. The results are compatible with any mechanism in which there is little development of charge in the transition state. This could be a transfer of hydride to the flavin N(5) position or a synchronous mechanism in which the α-C-H is formally abstracted as a H+ while the resulting charge is simultaneously neutralized by another event.

Prion protein NMR structure and species barrier for prion diseases

The structural basis of species specificity of transmissible spongiform encephalopathies, such as bovine spongiform encephalopathy or “mad cow disease” and Creutzfeldt–Jakob disease in humans, has been investigated using the refined NMR structure of the C-terminal domain of the mouse prion protein with residues 121–231. A database search for mammalian prion proteins yielded 23 different sequences for the fragment 124–226, which display a high degree of sequence identity and show relevant amino acid substitutions in only 18 of the 103 positions. Except for a unique isolated negative surface charge in the bovine protein, the amino acid differences are clustered in three distinct regions of the three-dimensional structure of the cellular form of the prion protein. Two of these regions represent potential species-dependent surface recognition sites for protein–protein interactions, which have independently been implicated from in vitro and in vivo studies of prion protein transformation. The third region consists of a cluster of interior hydrophobic side chains that may affect prion protein transformation at later stages, after initial conformational changes in the cellular protein.

Crystal structure of the RNA-dependent RNA polymerase of hepatitis C virus

We report the crystal structure of the RNA-dependent RNA polymerase of hepatitis C virus, a major human pathogen, to 2.8-Å resolution. This enzyme is a key target for developing specific antiviral therapy. The structure of the catalytic domain contains 531 residues folded in the characteristic fingers, palm, and thumb subdomains. The fingers subdomain contains a region, the “fingertips,” that shares the same fold with reverse transcriptases. Superposition to the available structures of the latter shows that residues from the palm and fingertips are structurally equivalent. In addition, it shows that the hepatitis C virus polymerase was crystallized in a closed fingers conformation, similar to HIV-1 reverse transcriptase in ternary complex with DNA and dTTP [Huang H., Chopra, R., Verdine, G. L. & Harrison, S. C. (1998) Science 282, 1669–1675]. This superposition reveals the majority of the amino acid residues of the hepatitis C virus enzyme that are likely to be implicated in binding to the replicating RNA molecule and to the incoming NTP. It also suggests a rearrangement of the thumb domain as well as a possible concerted movement of thumb and fingertips during translocation of the RNA template-primer in successive polymerization rounds.

Scanning mutagenesis of the putative transmembrane segments of Kir2.1, an inward rectifier potassium channel

Structural models of inward rectifier K+ channels incorporate four identical or homologous subunits, each of which has two hydrophobic segments (M1 and M2) which are predicted to span the membrane as α helices. Since hydrophobic interactions between proteins and membrane lipids are thought to be generally of a nonspecific nature, we attempted to identify lipid-contacting residues in Kir2.1 as those which tolerate mutation to tryptophan, which has a large hydrophobic side chain. Tolerated mutations were defined as those which produced measurable inwardly rectifying currents in Xenopus oocytes. To distinguish between water-accessible positions and positions adjacent to membrane lipids or within the protein interior we also mutated residues in M1 and M2 individually to aspartate, since an amino acid with a charged side chain should not be tolerated at lipid-facing or interior positions, due to the energy cost of burying a charge in a hydrophobic environment. Surprisingly, 17 out of 20 and 17 out of 22 non-tryptophan residues in M1 and M2, respectively, tolerated being mutated to tryptophan. Moreover, aspartate was tolerated at 15 out of 22 and 15 out of 21 non-aspartate M1 and M2 positions respectively. Periodicity in the pattern of tolerated vs. nontolerated mutations consistent with α helices or β strands did not emerge convincingly from these data. We consider the possibility that parts of M1 and M2 may be in contact with water.

A novel approach for the identification of protein–protein interaction with integral membrane proteins

Protein–protein interaction plays a major role in all biological processes. The currently available genetic methods such as the two-hybrid system and the protein recruitment system are relatively limited in their ability to identify interactions with integral membrane proteins. Here we describe the development of a reverse Ras recruitment system (reverse RRS), in which the bait used encodes a membrane protein. The bait is expressed in its natural environment, the membrane, whereas the protein partner (the prey) is fused to a cytoplasmic Ras mutant. Protein–protein interaction between the proteins encoded by the prey and the bait results in Ras membrane translocation and activation of a viability pathway in yeast. We devised the expression of the bait and prey proteins under the control of dual distinct inducible promoters, thus enabling a rapid selection of transformants in which growth is attributed solely to specific protein–protein interaction. The reverse RRS approach greatly extends the usefulness of the protein recruitment systems and the use of integral membrane proteins as baits. The system serves as an attractive approach to explore novel protein–protein interactions with high specificity and selectivity, where other methods fail.

Ras-catalyzed hydrolysis of GTP: a new perspective from model studies.

Despite the biological and medical importance of signal transduction via Ras proteins and despite considerable kinetic and structural studies of wild-type and mutant Ras proteins, the mechanism of Ras-catalyzed GTP hydrolysis remains controversial. We take a different approach to this problem: the uncatalyzed hydrolysis of GTP is analyzed, and the understanding derived is applied to the Ras-catalyzed reaction. Evaluation of previous mechanistic proposals from this chemical perspective suggests that proton abstraction from the attacking water by a general base and stabilization of charge development on the gamma-phosphoryl oxygen atoms would not be catalytic. Rather, this analysis focuses attention on the GDP leaving group, including the beta-gamma bridge oxygen of GTP, the atom that undergoes the largest change in charge in going from the ground state to the transition state. This leads to a new catalytic proposal in which a hydrogen bond from the backbone amide of Gly-13 to this bridge oxygen is strengthened in the transition state relative to the ground state, within an active site that provides a template complementary to the transition state. Strengthened transition state interactions of the active site lysine, Lys-16, with the beta-nonbridging phosphoryl oxygens and a network of interactions that positions the nucleophilic water molecule and gamma-phosphoryl group with respect to one another may also contribute to catalysis. It is speculated that a significant fraction of the GAP-activated GTPase activity of Ras arises from an additional interaction of the beta-gamma bridge oxygen with an Arg side chain that is provided in trans by GAP. The conclusions for Ras and related G proteins are expected to apply more widely to other enzymes that catalyze phosphoryl (-PO(3)2-) transfer, including kinases and phosphatases.

Close correspondence between the action spectra for the blue light responses of the guard cell and coleoptile chloroplasts, and the spectra for blue light-dependent stomatal opening and coleoptile phototropism.

Fluorescence spectroscopy was used to characterize blue light responses from chloroplasts of adaxial guard cells from Pima cotton (Gossypium barbadense) and coleoptile tips from corn (Zea mays). The chloroplast response to blue light was quantified by measurements of the blue light-induced enhancement of a red light-stimulated quenching of chlorophyll a fluorescence. In adaxial (upper) guard cells, low fluence rates of blue light applied under saturating fluence rates of red light enhanced the red light-stimulated fluorescence quenching by up to 50%. In contrast, added blue light did not alter the red light-stimulated quenching from abaxial (lower) guard cells. This response pattern paralleled the blue light sensitivity of stomatal opening in the two leaf surfaces. An action spectrum for the blue light-induced enhancement of the red light-stimulated quenching showed a major peak at 450 nm and two minor peaks at 420 and 470 nm. This spectrum matched closely an action spectrum for blue light-stimulated stomatal opening. Coleoptile chloroplasts also showed an enhancement by blue light of red light-stimulated quenching. The action spectrum of this response, showing a major peak at 450 nm, a minor peak at 470 nm, and a shoulder at 430 nm, closely matched an action spectrum for blue light-stimulated coleoptile phototropism. Both action spectra match the absorption spectrum of zeaxanthin, a chloroplastic carotenoid recently implicated in blue light photoreception of both guard cells and coleoptiles. The remarkable similarity between the action spectra for the blue light responses of guard cells and coleoptile chloroplasts and the spectra for blue light-stimulated stomatal opening and phototropism, coupled to the recently reported evidence on a role of zeaxanthin in blue light photoreception, indicates that the guard cell and coleoptile chloroplasts specialize in sensory transduction.

Gestão de embalagens vazias de agrotóxicos - logística reversa em pequenos municípios brasileiros: o caso do município de Bom Repouso, MG

O Brasil assumiu e mantém desde 2008 o primeiro lugar mundial em consumo de agrotóxicos e, tendo em vista este uso intensivo, é possível estimar a grande quantidade de embalagens vazias de agrotóxicos (EVAs) geradas todos os anos no país. Com a promulgação da Lei 9974 de 2000, a criação do Instituto Nacional de Processamento de Embalagens Vazias (inpEV) em 2001 e a implantação da Política Nacional de Resíduos Sólidos (PNRS) em 2010, a destinação adequada das EVAs por meio da logística reversa passou a ocorrer de maneira mais eficiente, chegando a um percentual de 94% de devolução em 2014 segundo dados do INPEV. No entanto, este processo foi originalmente concebido para atender grandes áreas rurais, com economias de escala que o favorecem, e portanto não está adequado à realidade de pequenas comunidades rurais, geralmente com infraestrutura precária e mais isoladas. Sendo assim, ainda é comum nos municípios de pequeno porte que essas embalagens sejam queimadas, armazenadas em locais não adequados ou simplesmente abandonadas no campo. Dessa forma, o objetivo deste estudo foi analisar a gestão de EVAs em pequenos municípios agrícolas brasileiros, com foco no processo de logística reversa, a partir de um estudo de caso. Este foi realizado em Bom Repouso, MG, município de 10500 habitantes, caracterizado pela produção de morango e batata, com mão de obra familiar e uso intensivo de agrotóxicos. As seguintes etapas de pesquisa foram desenvolvidas: entrevistas semiestruturadas com os atores da cadeia de logística reversa de EVAs; levantamento documental; análise comparativa entre a quantidade de embalagens vendidas e a quantidade total de embalagens devolvidas entre os anos de 2012-2013 e 2013-2014, bem como um levantamento dos agrotóxicos mais comercializados no município. Verificou-se que, no período 2012-2013, a quantidade de embalagens vendidas foi cerca de 30 vezes maior que a de embalagens devolvidas, e para o período entre 2013-2014 a mesma relação foi da ordem de 26 vezes. Além disso, a massa total de embalagens devolvidas corresponde a apenas 3,3% da massa de vendidas no período 2012-2013, e 3,8% para 2013-2014. Ou seja, aproximadamente 96% das embalagens comercializadas em ambos os períodos não foram devolvidas à central de Pouso Alegre. Além disso, há produtos extremamente tóxicos para a saúde e para o meio ambiente entre os 20 mais vendidos no município. Não estão estabelecidas políticas públicas municipais voltadas à gestão desses resíduos, como pode ser verificado através das entrevistas e do levantamento documental. Dificuldades relacionadas ao armazenamento das EVAs até o momento da devolução e seu transporte até a Central de Recolhimento foram apontadas pelos entrevistados como principais motivos para não cumprir suas responsabilidades e, consequente, descartar inadequadamente esses resíduos.

Dibujando la arquitectura olvidada: las defensas militares de la guerra de 1936-39 [PPT]

A decade before there was getting up the 'Atlantic Wall', there was executed a system of defenses along the Mediterranean coast in Spain (1936-39). The recovery of the same constructions (both of his graphical documents and of the constructed works that stay in foot) and his putting in value it can help to consolidate an own memory of the 20th century. This work considers to inventory, to measure and to draw the planes of these architectures to fix the memory that is diluted by the erosion of the time. The military pieces place in many borders: are these properly architecture? These are walking between two epochs: the one that perpetuates the epic acts in opposite to the one that shows the disasters in order that they do not forget. They are the most modern ruins of our history. In this process of reconstruction of the memory, there turns out to be crucial the graphical restitution. The drawing is a source of knowledge and demonstrates facts that were constructed.

Dibujando la arquitectura olvidada: las defensas militares de la guerra de 1936-39

Comunicación presentada en el XI Congreso Internacional de Expresión Gráfica Aplicada a la Edificación, APEGA 2012, Valencia, 29 noviemnre-1 diciembre 2012.