Analysis of Contributions to NO2 Ambient Air Quality Levels in Madrid City (Spain) through Modeling. Implications for the Development of Policies and Air Quality Monitoring

As environmental standards become more stringent (e.g. European Directive 2008/50/EC), more reliable and sophisticated modeling tools are needed to simulate measures and plans that may effectively tackle air quality exceedances, common in large cities across Europe, particularly for NO2. Modeling air quality in urban areas is rather complex since observed concentration values are a consequence of the interaction of multiple sources and processes that involve a wide range of spatial and temporal scales. Besides a consistent and robust multi-scale modeling system, comprehensive and flexible emission inventories are needed. This paper discusses the application of the WRF-SMOKE-CMAQ system to the Madrid city (Spain) to assess the contribution of the main emitting sectors in the region. A detailed emission inventory was compiled for this purpose. This inventory relies on bottom-up methods for the most important sources. It is coupled with the regional traffic model and it makes use of an extensive database of industrial, commercial and residential combustion plants. Less relevant sources are downscaled from national or regional inventories. This paper reports the methodology and main results of the source apportionment study performed to understand the origin of pollution (main sectors and geographical areas) and define clear targets for the abatement strategy. Finally the structure of the air quality monitoring is analyzed and discussed to identify options to improve the monitoring strategy not only in the Madrid city but the whole metropolitan area.

El centro comercial : una teoría tipológica : claves para el reconocimiento de una arquitectura ignorada

Existe en el panorama edificado un patrimonio construido que se reconoce como Centro Comercial. Un conjunto entendido, en sentido genérico, como familia arquitectónica que tiene características propias y específicas que la identifican. El objeto de la presente tesis doctoral consiste en argumentar que este conjunto constituye un nuevo tipo en el panorama de las tipologías arquitectónicas. Un tipo con entidad propia, que se conecta a una forma diferente de entender la idea de modelo. Un concepto que va más allá de la consideración tradicional del término. Modelo virtual. Este tipo, que surge de una estructura teórica que hemos denominado teoría tipológica, se constituye en una herramienta más para el estudio y el desarrollo proyectual de los espacios arquitectónicos, tanto del propio Centro Comercial como del conjunto de la disciplina arquitectónica, como referencia legítima. El presente trabajo de tesis se inicia con un bloque introductorio denominado Método. Definimos en él una metodología que hemos llamado emocional. Trata de la oportunidad de la tesis. Del porqué de un título que recoge la palabra ignorada. Del interés que suscita el asunto en el contexto del momento presente. Oportunidad e interés en base a una vida profesional dedicada al mundo del Centro Comercial y a la importancia del patrimonio elegido como objeto de estudio. También ha sido un aliciente detectar como las planificaciones del territorio y de los ámbitos de las relaciones colectivas no han sido capaces de integrar un resultado satisfactorio. Quizás por no considerar la complejidad de sus muchas facetas. En consecuencia, el texto busca la esencia del Centro Comercial como soporte para la crítica de su impacto en los nuevos escenarios de relación que la sociedad y el entorno físico imponen. Ámbitos donde los mecanismos históricos del asociacionismo tradicional han dado paso a otros, como el Centro Comercial, de exaltación del individualismo, pero demandados por una sociedad que se identifica con ellos. Espacios que, con Galbraith, existen por de la perversidad intrínseca del binomio consumo-producción que inducen la perplejidad. Ésta pasa a formar parte de la esencia del nuevo espacio comercial, como quedó de manifiesto en el Congreso de Minnesota de 1997, sobre el Centro Comercial. Una sociedad que ha girado hacia el logro material en términos de culto, ocasionando creciente valor significante del consumo. Razón última de la humanidad al decir de Rem Koolhaas. Culto que desemboca en la urgencia de alcanzar niveles de estatus y de identificación con el grupo. Dos parámetros que marcan la relación con el otro. Relación de comparación que excita la necesidad de posesión de objetos que llevaban a recrear en el consumidor la ilusión de ser especiales, de no pasar desapercibidos. El producto de consumo, el objeto, se eleva a la consideración de valor social. En el Centro Comercial se venden valores. Marketing de valores. El deseo del individuo, no la necesidad, queda involucrado en el proceso. La oportunidad y el interés de este estudio surgen además para aclarar el significado de un espacio que sirve al consumidor y a su entorno. Un significado que alcanza sentido, entre otros, por la aparición de un nuevo contexto tras la Segunda Guerra Mundial. Suburbanización, automóvil, nivel de vida, un nuevo papel de la mujer. La Era del Consumo. Una era que genera la paradoja de un individuo con autodeterminación y autosuficiencia crecientes, en un mundo cada vez más condicionado y controlado por dinámicas de ofertas mediatizadas e ilimitadas. La arquitectura en este contexto ha de juzgarse por su relación con un hombre contemporáneo que camina hacia una progresiva excepcionalidad. Cerramos la presentación justificando el término ignorada que aparece en el Título de la Tesis. Subrayamos la escasa existencia de estudios estructurados que relacionen Centro Comercial y arquitectura. Realidad que se constata partiendo del libro de Pevsner, Historia de las Tipologías Arquitectónicas, de 1976. Analizamos también la aceptación que el Centro Comercial, valorado en algunos ámbitos como arquitectura populista. Primer Capítulo, los antecedentes. Se propone un recorrido histórico por la arquitectura comercial de todos los tiempos. El Documento busca concretar las características de la arquitectura comercial a lo largo de la historia para determinar relaciones entre ella y el Centro Comercial. Estas correspondencias van a permitir contestar la pregunta retórica que nos hacemos al inicio del Capítulo: ¿Es la arquitectura del Centro Comercial una arquitectura subsidiaria, o tiene personalidad propia independiente de la del mundo comercial? Con Zevi40, queremos dejar constancia que la relación arquitectura-historia es imprescindible en la tarea proyectual. Un análisis novedoso solo es posible desde la búsqueda de unas raíces auténticas. Así mismo, con Guy de Maupassant, estamos convencidos que “La arquitectura, a través de los siglos, ha tenido el privilegio de dar un símbolo a cada una de las épocas, de resumir con un pequeñísimo número de monumentos típicos, el modo de pensar, de soñar de una raza y de una civilización”. Este recorrido se inicia interpretando la arquitectura comercial minoica. Llegaremos, paso a paso, hasta el siglo XX, los grandes itinerarios comerciales y el Centro Comercial. Se descubren una serie de invariantes que permiten comparar y extraer conclusiones. Resulta novedoso constatar que el Centro Comercial nace para dar respuesta al hombre contemporáneo en su afán consumista. También lo es la rápida implantación y evolución del Centro Comercial en un corto periodo de sesenta años frente al ritmo sosegado de otras soluciones comerciales. Novedad es ver como el comercio y sus arquitecturas nacen abiertos, bajo tenderetes y el Centro Comercial se presenta cerrado. Así mismo, las referencias sacras constituyen un elemento de novedad para la reflexión, en un contexto materialista. Y tantas otras. En Minos, la óptica comercial ofrece otra visión de su legendaria cultura. ¿Palacio o plataforma logística?, gestión centralizada del intercambio, ¿vida pública y vida privada? Así, hasta llegar al siglo XIX. Sus las galerías y sus pasajes acristalados concebidos en primera instancia como medida de recomposición urbana. Espacio entendido desde lo público-privado, desde lo interior-exterior, desde el dentro-afuera, desde lo cerrado-abierto. Con los nuevos mercados de abastos, representan una revolución en el ámbito de la funcionalidad, de la máquina, de la gestión moderna y de las relaciones del ciudadano con la ciudad apoyado en base al intercambio de bienes de consumo. El Centro Comercial es heredero de esta transformación. El Gran Almacén, por otra parte, es el reflejo de otra gran revolución. La que va ligada a la producción y comercialización en masa, las comunicaciones, el precio fijo y el aumento del nivel de vida. El Centro Comercial reinterpreta estas situaciones insertándolas en un nuevo modelo de gestión. En el siglo XX, maestro en técnicas de venta, aparece el hipermercado. El Híper, con su carga de pedagogía, se incorpora al esquema orgánico del Centro Comercial. La tienda en si misma constituye la pieza base de dicho puzle orgánico. Es en esta época cuando la tienda empieza su despegue autónomo como arquitectura, aportando su experiencia. Tras ello, llegamos a las grandes rutas comerciales, que proponemos como metáfora del shopping. Cerramos el capítulo concluyendo con Eugenio Ferrer que “si establecemos una relación entre el espacio y el capitalismo, entonces podemos inferir que los espacios del consumo de masas (ECM) son configuraciones nuevas respecto al pasado (...), pero el sistema que lo introduce no lo es del todo”. Segundo Capítulo. Búsqueda de claves que permitan el reconocimiento del Centro Comercial. Llegados a este punto y con la perspectiva del tiempo cabe preguntarnos, ¿qué entendemos pues por Centro Comercial? ¿Cómo lo percibimos? Abordamos ahora el problema de la percepción del Centro Comercial y su significado. Además de constituirse en símbolo, referencia siempre presente, la eficacia del Shopping es una de las principales causas de su poder de atracción. El Centro Comercial resulta de la síntesis de la revolución del consumo y de la revolución de la gestión. Espacio eficaz del entretenimiento como destino. El usuario resuelve su vida cotidiana dentro de un hábitat que considera propio y que se entiende como el lugar hiperbólico de la transacción comercial. Espacio de la abundancia para el disfrute. Una arquitectura involucrada en esta eficacia. Su sentido del lugar no es otro que el Shopping, que se desenvuelve de forma análoga en todas partes. El hogar del consumidor. Las nuevas catedrales. Las catedrales del consumo. Destino místico. Lugar de peregrinación para el consumidor fervoroso. Espacio sagrado que integra al usuario en la cultura dominante. Cultura del consumo. Templos, donde el tiempo ha perdido su sentido. Paraíso. Un espacio donde la altura, la luz natural, la dimensión general refuerzan el carácter sacro de un espacio para una nueva religión laica. Un espacio seguro, protegido que nos acerca a ideas como la de útero materno, con su carga de calidez y de significación erótica aplicado al encuentro compra-comprador y, ligado a ello, la literatura especializada habla del Centro Comercial como nave espacial hiperesterilizada o de agujero negro que absorbe la energía cultural. Más allá, la simulación, donde se percibe un simulacro de ciudad. Simulacro coherente con todo lo que el Shopping desencadena a su alrededor. El lugar de los sueños, de la fantasía. Aquí los productos se han metamorfoseado en fetiches, en significados. Televisión tridimensional, donde el usuario actúa guiado por una pulsión similar al zapping. Espacio lúdico de la fascinación por comprar o de imaginar que se compra. Espacio de simulaciones que llevan a la ensoñación. Un nuevo lugar que sustituye al espacio cotidiano, con el señuelo de la protección, en un contexto imaginariamente público. Espacio de la hiperrealidad donde no se distingue la realidad de la fantasía, donde tras episodios de confusión y desconcierto, se borran las fronteras con lo imaginario. Espacio mágico, escenario del gran espectáculo del consumo, controlado milimétricamente, al modo de la visita a un gran monumento, que ha de sobrevivir a los tiempos para dar testimonio de nuestro momento. Un icono, un símbolo que transmite un mensaje, que solo el consumidor es capaz de interpretar. Una agitada mezcla, sin precedentes, de percepciones que hablan de perplejidad y asombro ante el fenómeno del Centro Comercial, su espacio y las reacciones del hombre contemporáneo. ¿Cuáles serían las claves que nos permitirían reconocer la calidad esencial de un Centro Comercial, en esta concurrencia de perplejidades? Primero, la función de servicio. Un espacio donde ocurren muchas más cosas que lo obvio, que la compra. Un edificio que se involucra con el entorno de la mano de lo inesperado, la sorpresa y las expectativas. Esta vocación de servicio conecta Centro Comercial y naturaleza arquitectónica. Función que sugiere percepciones ligadas a la experiencia de compra. Organismo que vive y late al unísono con su visitante, colocándose al servicio de sus necesidades, de su afán de consumo, del que vive. Segundo, la comunidad servida. El Centro comercial sirve a una sociedad concreta. La sociedad consumista. Una nueva sociedad que se identifica con el edificio desarrollando un sentido de comunidad al nivel de sus deseos. Esta comunidad que se configura a su alrededor, constituye el activo más poderoso para el éxito de su realidad cotidiana y de su futuro. Tercero, un compromiso de carácter holístico. La economía de la experiencia aplicada al afán consumista de una sociedad identificada con su Centro, da lugar a una experiencia holística planificada. Diseño emocional. Colaboración para el éxito de un conjunto de establecimientos comerciales que participan en la aventura espacio-comercial del Centro Comercial. Ellos son los inductores primarios del consumo. Pero esta colaboración tiene su culminación en la amplificación del mensaje, como un inmenso altavoz, que proviene de la unidad configurada por todos ellos. El reflejo de esta amplificación de mensajes, desde la poderosa unidad constituida, es el aumento de la rentabilidad, fin último de la operación. Cuarto, la forma a través de una identificación de carácter gestáltico. Desde la lectura gestáltica que hacemos de la unidad holística, se advierte una poderosa capacidad de comunicación del sistema con su contexto. Centro y entorno se tornan entonces cómplices que complementan sus realidades. El Centro Comercial, arquitectura estructurada como sistema se percibe –ha de percibirse- como forma unitaria que procede de una mirada de raíz gestáltica que continuamente la recompone desde una óptica espacial y física, ligada a la experiencia individual. Esta unidad formal, más allá de la forma real, se constituye en esencia de de su arquitectura. Quinto, el Centro Comercial como sistema. Un sistema soportado por la Teoría General de Sistemas. La consideración del Centro Comercial como sistema es consecuencia de su estructura holística. El todo no se comporta como la suma de las partes y estas no lo hacen como lo harían en solitario. De aquí surge la necesidad de diálogo permanente entre la comercialización –proceso de incorporación de partes- y su traducción al mundo del diseño –proceso de articular arquitectónicamente las partes. Como sistema así configurado, el Centro Comercial se inserta en el paradigma contemporáneo, lo que genera realidades duales que no son excluyentes y reacciones de perplejidad e incertidumbre que el sistema corrige con su capacidad de autorregulación. Aparece también el espectador cuántico, el visitante, el consumidor, que interactúa con el sistema. Desde las herramientas que nos aporta la idea de sistema complejo, afrontamos el Mix Comercial -en definitiva la eficaz localización de las piezas en orden a sus relaciones y al organismo resultante- y su incidencia en la arquitectura que estamos concibiendo. Una arquitectura impredecible por lo mutable, que se erige en reto de la operación y del diseñador. Diseño que, de la mano del concepto de sistema se convierte en herramienta a mayor gloria de la operación global. El debate del estilismo no será más que el resultado del análisis en busca del éxito de esta operación. Sexto, una arquitectura de la negociación. Negociación como mecanismo proyectual y como resultado. La solución de proyecto nunca resulta evidente en el Centro Comercial como consecuencia de lo imprevisible del propio proceso de configuración. Su concreción solo puede ser fruto del compromiso de todos los agentes por conseguir el objetivo de la operación. Esto se consigue desde el equilibrio de intereses. Comerciales y de diseño. Un compromiso con la negociación y una negociación íntimamente ligada a la coordinación. Séptimo, el espacio y el tiempo. El debate espacio-tiempo condiciona y estructura la percepción del Centro Comercial. Introducimos conceptos como cronotopo –el instante y el lugar donde ocurre algo- y paradoja –incoherencia de la relación causa efecto- que sitúan el vínculo entre el tiempo y el espacio del Centro Comercial en un contexto de Shopping. En el discurrir paralelo del tiempo histórico –el tiempo de fuera- y del tiempo interior, el de dentro del Centro Comercial –tiempo presente o intemporalidad-, se produce el triunfo social del Centro Comercial que se traduce en haber sabido resolver en el espacio y en el tiempo las paradojas postmodernas del hombre contemporáneo. Octavo, de lo global. Globalidad que no es ajena a lo local. Una arquitectura que insertada en la dinámica de una economía de ámbito mundial, refleja las contradicciones que ella impone, fundamentalmente en los procesos de inclusión y exclusión, afectando de manera decisiva al debate de lo local, que el Centro Comercial debe incorporar como herramienta ineludible de reconocimiento. Terminamos el capítulo segundo manifestando como estas ocho claves, asumidas en su conjunto, confirman que el Centro Comercial puede aparecer como un todo conceptual cohesionado, pasando a formar parte de una familia arquitectónica coherente, cuya estructura funcional somos capaces de establecer. El Capítulo Tercero presenta con detalle la figura del arquitecto Víctor Gruen, creador reconocido del moderno Centro Comercial. Presentamos su trayectoria profesional observando como las diferentes claves analizadas en capítulos anteriores van apareciendo de manera natural a lo largo de ella "Victor Gruen may well have been the most influential architect of the twentieth century." Malcolm Gladwell. The Terrazo Jungle. Fifty Years Ago, The Mall Was Born. America Never Would Be The Same. 2004. In The New Yorker. Hombre complejo, conflictivo. Las paradojas a lo largo de su carrera fueron notables. Sin embargo siempre apareció como un hombre recto. Garret Eckbo, reconocido paisajista y colaborador de Gruen en el Centro Comercial peatonal del Centro urbano de Fresno, California, se asombraba de que alguien como Gruen hubiese sido capaz de combinar cortesía y humanidad en la carrera de ratas (sic) en la que se convirtió el universo de la comercialización americana y mantener la integridad542. Philip Johnson, en 1962, ponderando las muchas habilidades de Gruen manifestó que no estaba seguro si alguno de ellos, arquitectos artistas, hubiera sido capaz de hacer lo que Gruen hizo. A más, sobre Gruen, manifestó lo siguiente: "... El va más allá de la creación de un bello edificio. En jugar con la gente y sugerir lo que tienen que hacer, es un maestro. Y obtiene buenos resultados como hace la escultura. El suyo es un arte cívico, un sentido cívico.... Él es capaz de sentarse y poner cosas juntas. No es pomposo ni vano. Yo no me reuniría con él para hablar del diseño (de edificios). Víctor siente que cuando se habla de diseño se está ignorando todo el contexto... Su arquitectura es poderosamente limpia, no vuela la fantasía. Pero cuando te haces con su complejidad, ves que has descubierto algo más allá del diseño.... No puedes decir que haya alguien como él. La arquitectura tiene la suerte de tenerle como arquitecto..." Philip Johnson. Article in Fortune Review. 1962. El Documento de tesis cierra la visión de Gruen realizando un recorrido por los diferentes capítulos de su libro esencial, Shopping Towns Usa: The Planning of Shopping Centers. Solo su índice resulta un monumento al proceso de gestación del Centro Comercial. El libro, aquí simplemente mostrado en su estructura básica como un silencioso testigo, es la referencia canónica los Centros Comerciales contemporáneos, desde su aparición. . El Cuarto Capítulo del Documento de tesis es una recapitulación del trabajo anterior, en el que se sintetizan los conceptos de función y estilo relacionados con el Centro Comercial, se define en qué consiste cada uno de ellos y como, a partir de ahí, podemos afirmar que nos encontramos ante un tipo arquitectónico nuevo en el panorama de la disciplina arquitectónica. Terminamos el Capítulo integrando el Centro Comercial con un cuerpo teórico de referencias que se remiten a un tipo arquitectónico concreto y particular, acogiendo su singularidad como fenómeno arquitectónico autónomo. Como Conclusión de la tesis, resultado de todo lo anterior es decir, como consecuencia de la integración de un torrente de percepciones e intuiciones en un cuerpo teórico de referencias, deducidas de la existencia de unas claves que estructuran y penetran la esencia del singular modelo estudiado, haciéndolo detectable y seductor, resultan las características de un tipo arquitectónico con entidad propia que ordena, orienta y supedita la realidad y la existencia de esta nueva arquitectura. Una arquitectura nunca antes definida como tal, en el panorama tipológico de la disciplina. Teoría tipológica para una nueva arquitectura, que hemos ido proponiendo a lo largo del trabajo y que es coherente con los diferentes parámetros que se han analizado. Un conjunto edificado que, desde el estudio de sus claves esenciales y de sus invariantes perceptibles, aparece ahora más cercano, más familiar. Tanto que es posible destilar desde este conocimiento cercano e íntimo, una síntesis útil como referencia proyectual y como referencia para las grandes cuestiones que preocupan al discurrir del debate arquitectónico y sus ideas. El debate de la disciplina. El objeto de esta tesis, que consistía en establecer que el conjunto edificado que conocemos como Centro Comercial se constituye en un nuevo tipo en el panorama de las tipologías arquitectónicas, entendido el Centro Comercial en sentido genérico como familia arquitectónica con características propias y específicas que la hacen autónoma y reconocible, queda a nuestro juicio argumentado y justificado. ABSTRACT Within the frame of the built heritage there is a construction that is recognized as Shopping Center. An ensemble understood as an architectural family with its own specific recognizable characteristics. This thesis aims to explain that this building complex constitutes a new type in the panorama of architectural typology. This typology, with its singular identity, is connected to a way of understanding the idea of the model beyond an orthodox conception of the term understood as virtual model. This typology comes from a theoretical structure that we called typological theory, and it serves as yet another tool to reference the study and development design of the architectural spaces. In this first section, the Method, we emotionally explore the opportunities of this thesis. Why this typology has been ignored and the interest this work has in the present moment. An opportunity and an interest explained from an experience of a life dedicated to the world of Shopping Centers. The text then introduces the need for a rigorous knowledge of the Shopping Center’s essence in order to understand its impact in the frame of a new society and a new physical environment. A frame time where the historical mechanisms of association of civic community have given way to other gathering spaces like the Shopping Center, which encourages individualism, but is demanded by a society that relates to them. Spaces that, according to Galbraith, are a result of the intrinsic perversity of the unstoppable movement of the wheel of consumption-production. A society that has turned to worship of material achievement. Worship that provokes the appearance of an increasing value of consumption, according to Koolhaas, the only goal of humanity. Worship that ends in the need to reach certain status levels in the plane of a permanent comparison where the need of possession excites the consumer and gives them the illusion of being special. The product of consumption rises up to consideration of social values, entering a dynamic of marketing values, not only objects, but the desire of the individual remains. The study appears also to clarify the meaning of a space that serves the consumer and its context. A meaning that makes sense with the appearance of the suburbanization, the massive utilization of the car, the increase of living standards and the new role of women in the society after the Second World War, giving rise to the Age of Consumption. A world now determined and controlled by media and unlimited offers, where it’s necessary to place them in the context of the ordinary. An architecture that has to be judged precisely for its relation with this specific contemporary man. This section ends justifying the term ignored that appears in the Title of the Thesis, considering it in relation with the lack of studies structured about the Shopping Center and its architecture, drawing from Pevsner's work, A history of building types, 1976. Finally, the Shopping Center is analyzed with the most critical of thoughts, which considers it as populist architecture. The First Chapter, Precedents, proposes an historical tour of the commercial architecture throughout history. The Document looks to place on record the characteristics of the commercial architecture to set the connection between them and the Mall itself. These correspondences are going to allow us to answer the rhetorical question: is it the architecture of the Shopping Center a subsidiary architecture, or does it have its own personality independent from that of the commercial world? The reason of this historical search, citing Bruno Zevi, is that it is indispensable to establish the relationship between architecture and history, understating that an analysis is only possible when researching for their roots. Moreover, according to Guy de Maupassant, we are sure that architecture has had the privilege, across the centuries, of symbolizing as it were each age(…), through the harmony of lines and the charm of ornamentation all the grace and grandeur of an epoch. This historical reading, inseparable from a consistent design action, begins interpreting the commercial architecture of the Minoan to the 20th Century. Though this analysis of the big commercial itineraries and the study of the Shopping Center itself. A reading where we have found a series of constants that make it possible to draw conclusions from this comparison. The Mall appears to give response to the needs of a consumerist society. Comparing to the calm pace of the evolution of other commercial solutions, it is relevant its fast implantation and evolution in a short period of sixty years. Though via different solutions, the commercial spaces are considered taking into account the public-private relation, the interior-exterior, the inside-out, the closed-opened. Through that, the 19th century galleries and the food markets represent a revolution in functionality, in the machine, the modern management and the relations of the citizen within the city. All of this, the Mall inheritor feels. Likewise, the Department Store is the reflection of another great revolution. Production and commercialization en-mass, communications, the fixed price and increase of the living standard. The Mall reinterprets these situations inserting them in a new model of management. Already in the 20th century Mall and mass technologies of sale, the hypermarket is enthusiastically incorporated into the configuration of this organic scheme, constituting the base of one of the models, the French, that will be highly developed in the European continent. The shop itself constitutes, on the other hand, the key piece that completes the puzzle of the Mall and is in this epoch when it starts taking off as architecture, has an autonomous character. After all this, finally, we come to the big commercial routes, which we propose as metaphor of the shopping. Citing Eugenio Ferrer we can conclude that “If we establish a relation between the space and the capitalism, then we can infer that the spaces of the consumption of masses (ECM) are new configurations with regard to the past (...), but the system that introduces it it is not completely” Now we arrive at this point and with the perspective of time it is necessary to ask us, what do we understand about the Shopping Center? How do we perceive it? The second Chapter approaches the problem of the perception of the Mall and from this it is possible to detect and to identify key drivers that orientate the architectural comprehension of the space. The efficiency of the Shopping Center is its main power of attraction. A world that has ensued from the synthesis of the revolution of consumption and management. An effective cavern-like place of entertainment where the user, the consumer, the postmodern man solves his daily life inside a considered habitat. The hyperbolic place of commercial transaction. An abundance of space, that makes us perceive it as destination of entertainment. An architecture has evolved this efficiency, where the sense of place is at one with the sense of the Shopping. The home of the consumer. The new cathedrals. The cathedral of consumption. The place of peregrination for the fervent consumer. A sacred space that integrates the user in the dominant culture. A temple, where time itself has stopped existing. In this paroxysm, an expression of the Garden of Eden or Paradise itself. A space where the height, the natural daylight and the spatial dimension reinforce the sacred character of a new lay religion. Another common perception is that of a protected area, which leads to metaphors and considerations that suggest the idea of maternal womb, with its weight of erotic meaning, referring to the encounter of the shopper making a purchase. The literature also tells us about its perception as a sterile space capsule, a black hole that absorbs all cultural energy. Likewise, a world simulation where a mock city is perceived at first instance. Consistent with all that shopping triggers inside. A city, a space conceived as a place of dreams, fantasy, where the products have been metamorphosed into fetishes. Entertaining a television, three-dimensional television, where the user acts guided by a drive similar to zapping. A play area where the latest fascination is in the act of buying. Space simulations that unite and transcend creating atmospheres that lead to reverie. A new space replacing the daily space with the lure of safe space in a public context. A hyper-reality space with reality and fantasy, where borders are erased with imaginary episodes of confusion and bewilderment, without distinction. The charm and fascination of a space that reads like magic. The magic of a space which is defined as stage extravaganza, the large theatre, the consumer surveys in the fine control mode in which you visit a national monument. The shopping center has to survive the times to be a testimony of our time. An icon, a symbol that conveys a message, the message reads ‘consumer’. In short, a Shopping Center is a mix of unprecedented insights that speak of a widespread phenomenon of bewilderment. Its space and the reactions of contemporary man unfold in it like a fish in water. What are the key aspects which allow us to recognize the essential quality of a shopping center in this concurrence of perplexities? First, we want to record a service function of space much deeper than the immediately obvious, i.e. a purchase occurs. A building that appears to be involved with the environment and its people from the hand of the unexpected circumstances; surprise and attention. And that, in turn, also involves the visitor beyond the purchase. This dedication to service closely links the mall with its architectural nature. It is not the function of a lifeless machine. It is a feature that suggests unsuspected perceptions linked to the purchase, which speaks of an organism that lives and breathes in unison with the visitor. Second, in addition to the vocation of service-oriented desire for consumption, the Mall environment serves a particular society - The consumer society. A new society which relates to building a sense of community developed to the level of their desires. This community also constitutes the most powerful asset to the success of the daily life of the Shopping Center. Third, we emphasize that the so called economy of the experience is combined with the consumer zeal of a company that is identified by the Shopping Center. It connects to form a holistic and planned experience. This experience takes shape in the entity that ensues from the premeditated association and synergy, in the sense of a collaboration for success. A set of concrete commercial and independent establishments, take part in the spatial and commercial adventure that is the Mall and they are the instigators of the consumption. This holistic behavior finds culmination in the amplification of a claim that becomes unitary, like an immense force that leads to an increased profitability to all the levels. Consummation is a reference of one human being overturned in an architecture assimilated into a legitimate, emotional design with stability. A holistic quality is born of the essence of the building - and by virtue of the Conditions of Alexander, Christopher Alexander, determines the system condition of the Shopping Center. Fourth, we propose to establish what character the Mall will form when joined with the concept of its typology. This is going to allow the architectural work to be formed. As a result of the holistic structure that we see, the Mall is perceived as a system whose parts have their own function, justifying their existence in the ecosystem. Across a gestalt there is a powerful capacity of communication between the system and its context. We visualize on the one that stands out our building, turning both, Center and environment, in accomplices of a few special relations who complement each other in his realities. This relationship within a complex and diverse environment gives the Mall a range of unique spatial perceptions, the result of disparate experiences, which because of its root origin of gestalt, are integrated into a unified and coordinated manner fully intelligible and organized. This is the final formal essence of the Shopping Center. We can conclude here that the Mall as architecture is a structured system and should be perceived as a unit both from a physical and spatial perspective as this is the essence of its architecture. Fifth, the Mall as a system. A system which is being supported by a broad theoretical corpus, the General Systems Theory, which offers sufficient methodology to descend into consideration and give an enlightened conclusion on the overall understanding of the Mall. Consideration of the Mall as a system is a result of its holistic structure. The whole does not behave like any of the parties and they do not behave the way they used to before belonging to the whole, because they inhibit many of their qualities to their advantage. It arises the need for an ongoing dialogue between marketing processes and its translation into the physical world, the design. The system generates multiple perceptions to be integrated into a body which is to be understood as unitary. As a system, the Mall is inserted into the contemporary paradigm, creating dual realities that are not exclusive and are reactions of uncertainty that the system be properly designed at all levels, faced with their ability to self-regulate. Likewise, considering the visitor, the customer, like the quantum spectator who interacts with the system permanently. Moreover, a complex system confronts us with the Commercial mix, the effective location of parts in order to relate to the body and its importance in the architecture we are conceiving. Unpredictable architecture, which stands as the challenge of the operation and the designer. Design that becomes the tool of the system to create success for the overall operation. The discussion of the styling is merely the result of analysis that also seeks the success of the system, i.e. the styling should send the right message for the environment to ensure its survival. Sixth, the idea of negotiation as an architecture project, a mechanism inherent to the status of the proposed system. The project solution is never evident at the Shopping Center because of the unpredictability of the process itself. It can only be the fruit of the commitment of all stakeholders to achieve the objective of the operation. This is achieved from the balance of interests, of commercial and design. A commitment to negotiation and a negotiation linked to coordination. The pursuit of stability is key, as instability is always present and constantly requires strategies to build the object you are configuring. Seventh, proposes space-time itself as a circumstance that determines and structures the perception of the Mall in a singular way. We introduce concepts as chronotope and paradox to help us place the relationship between time and space within the Mall in the context of shopping. A consequence of the parallel flow of historical time - the time outside - and the time inside the Mall, the big shopping center formula is precisely that of having the feeling of timelessness in the space. The social triumph of the mall is the ability to resolve in space and in time all postmodern paradoxes and, beyond that, of contemporary man, condensing into a small space and time an enormous amount of cultural symbols, often contradictory, but they attract the practice of consumerism. Eighth, global level. Globalization which doesn’t ignore the local level. Architecture that is inserted into the dynamics of a global economy, reflects the contradictions that it imposes, mainly in the processes of inclusion and exclusion. Inclusion and exclusion affect the debate of the local level, which the Mall must incorporate as an unavoidable tool of recognition. The eight fundamental principles, when applied as a whole, confirm that the built heritage, which corresponds to the general Mall idea, can be presented as a cohesive conceptual whole. This becomes part of a coherent architectural family, whose functional structures are able to be established. The Third chapter presents in a detailed way the figure of the architect Victor Gruen, recognized as the creator of the modern Mall. Studying his professional experience, it is shown how the different keys analyzed in previous chapters are appearing in a natural way. "Victor Gruen may well have been the most influential architect of the twentieth century." Malcolm Gladwell. The Terrazo Jungle. Fifty Years Ago, The Mall Was Born. America Never Would Be The Same. 2004. In The New Yorker. He was a complex, troubled man and the paradoxes along his career were notable. Nevertheless, always he appeared as a straight man. Garret Eckbo, the recognized landscape painter and collaborator of Gruen was astonished how Gruen had been capable of combining comity and humanity in this career of rates, into that the American commercialization turned, Johnson, in 1962, weighting many Gruen's skills demonstrated that he was not sure if anyone of them, architects artists, had been capable of doing what Gruen did. He goes beyond just the creation of a beautiful building. In playing on people and suggesting what they ought to do, he is a master. (…) his architecture is clean - hardly architecture, no flights of fancy. But when you get hold of its complexity, you've got something beyond the design... You can't say there's someone like him. Architecture is lucky to have him as an architect." Philip Johnson. Article in Fortune Review. 1962. The Document of the thesis closes with Gruen's vision of realizing a tour through the different chapters of his essential book, Shopping towns USA: The Planning of Shopping Centers. It’s mere index turns out to be a monument to the process of the gestation of the Mall. The book, simply acted in its basic structure as a silent witness, as the canonical reference for the contemporary Malls. The Fourth Chapter of the Document of the thesis is a recapitulation of the previous work, which synthesizes the concepts of function and style related to the Shopping Center, and clearly defines how they are defined so we can conclude that we have found an architectural new type in the panorama of the architectural discipline. Therefore, the Conclusion of the thesis integrates this development in a theoretical body of references that relate to an architectural specific and particular type, which receives the singularity of the Shopping Center as an architectural independent phenomenon as it has tried to demonstrate from the beginning of the work. To conclude, as a result of the integrative process and the development of the theoretical body of references, the essential characteristics of the order and concept of the architectural typology form the existence of a new architecture; architecture never before defined as such, in the theoretical typology of the discipline. A theoretical typology for a new architecture is proposed throughout the discussed research and forms a conclusion of the different parameters that have been analysed. As a building complex, from the study of the essential characteristics and of the perceptible constants, the typology is more clearly defined and thus, becomes a useful tool and precedent for the consideration of the discipline. The thesis then justifies how the building complex known as Shopping Center constitutes a new type of architectural typology.

Quantitative fine-structural analysis of olfactory cortical synapses

To determine the extent to which hippocampal synapses are typical of those found in other cortical regions, we have carried out a quantitative analysis of olfactory cortical excitatory synapses, reconstructed from serial electron micrograph sections of mouse brain, and have compared these new observations with previously obtained data from hippocampus. Both superficial and deep layer I olfactory cortical synapses were studied. Although individual synapses in each of the areas—CA1 hippocampus, olfactory cortical layer Ia, olfactory cortical area Ib—might plausibly have been found in any of the other areas, the average characteristics of the three synapse populations are distinct. Olfactory cortical synapses in both layers are, on average, about 2.5 times larger than their hippocampal counterparts. The layer Ia olfactory cortical synapses have fewer synaptic vesicles than do the layer Ib synapses, but the absolute number of vesicles docked to the active zone in the layer Ia olfactory cortical synapses is about equal to the docked vesicle number in the smaller hippocampal synapses. As would be predicted from studies on hippocampus that relate paired-pulse facilitation to the number of docked vesicles, the synapses in layer 1a exhibit facilitation, whereas the ones in layer 1b do not. Although hippocampal synapses provide as a good model system for central synapses in general, we conclude that significant differences in the average structure of synapses from one cortical region to another exist, and this means that generalizations based on a single synapse type must be made with caution.

The design, computer modeling, solution structure, and biological evaluation of synthetic analogs of bryostatin 1

The bryostatins are a unique family of emerging cancer chemotherapeutic candidates isolated from marine bryozoa. Although the biochemical basis for their therapeutic activity is not known, these macrolactones exhibit high affinities for protein kinase C (PKC) isozymes, compete for the phorbol ester binding site on PKC, and stimulate kinase activity in vitro and in vivo. Unlike the phorbol esters, they are not first-stage tumor promoters. The design, computer modeling, NMR solution structure, PKC binding, and functional assays of a unique class of synthetic bryostatin analogs are described. These analogs (7b, 7c, and 8) retain the putative recognition domain of the bryostatins but are simplified through deletions and modifications in the C4-C14 spacer domain. Computer modeling of an analog prototype (7a) indicates that it exists preferentially in two distinct conformational classes, one in close agreement with the crystal structure of bryostatin 1. The solution structure of synthetic analog 7c was determined by NMR spectroscopy and found to be very similar to the previously reported structures of bryostatins 1 and 10. Analogs 7b, 7c, and 8 bound strongly to PKC isozymes with Ki = 297, 3.4, and 8.3 nM, respectively. Control 7d, like the corresponding bryostatin derivative, exhibited weak PKC affinity, as did the derivative, 9, lacking the spacer domain. Like bryostatin, acetal 7c exhibited significant levels of in vitro growth inhibitory activity (1.8–170 ng/ml) against several human cancer cell lines, providing an important step toward the development of simplified, synthetically accessible analogs of the bryostatins.

Altered 3′-terminal RNA structure in phage Qβ adapted to host factor-less Escherichia coli

The RNA phage Qβ requires for the replication of its genome an RNA binding protein called Qβ host factor or Hfq protein. Our previous results suggested that this protein mediates the access of replicase to the 3′-end of the Qβ plus strand RNA. Here we report the results of an evolutionary experiment in which phage Qβ was adapted to an Escherichia coli Q13 host strain with an inactivated host factor (hfq) gene. This strain initially produced phage at a titer ≈10,000-fold lower than the wild-type strain and with minute plaque morphology, but after 12 growth cycles, phage titer and plaque size had evolved to levels near those of the wild-type host. RNAs isolated from adapted Qβ mutants were efficient templates for replicase without host factor in vitro. Electron microscopy showed that mutant RNAs, in contrast to wild-type RNA, efficiently interacted with replicase at the 3′-end in the absence of host factor. The same set of four mutations in the 3′-terminal third of the genome was found in several independently evolved phage clones. One mutation disrupts the base pairing of the 3′-terminal CCCoh sequence, suggesting that the host factor stimulates activity of the wild-type RNA template by melting out its 3′-end.

Structure of the imprinted mouse Snrpn gene and establishment of its parental-specific methylation pattern

The mouse Snrpn gene encodes the Smn protein, which is involved in RNA splicing. The gene maps to a region in the central part of chromosome 7 that is syntenic to the Prader–Willi/Angelman syndromes (PWS-AS) region on human chromosome 15q11-q13. The mouse gene, like its human counterpart, is imprinted and paternally expressed, primarily in brain and heart. We provide here a detailed description of the structural features and differential methylation pattern of the gene. We have identified a maternally methylated region at the 5′ end (DMR1), which correlates inversely with the Snrpn paternal expression. We also describe a region at the 3′ end of the gene (DMR2) that is preferentially methylated on the paternal allele. Analysis of Snrpn mRNA levels in a methylase-deficient mouse embryo revealed that maternal methylation of DMR1 may play a role in silencing the maternal allele. Yet both regions, DMR1 and DMR2, inherit the parental-specific methylation profile from the gametes. This methylation pattern is erased in 12.5-days postcoitum (dpc) primordial germ cells and reestablished during gametogenesis. DMR1 is remethylated during oogenesis, whereas DMR2 is remethylated during spermatogenesis. Once established, these methylation patterns are transmitted to the embryo and maintained, protected from methylation changes during embryogenesis and cell differentiation. Transfections of DMR1 and DMR2 into embryonic stem cells and injection into pronuclei of fertilized eggs reveal that embryonic cells lack the capacity to establish anew the differential methylation pattern of Snrpn. That all PWS patients lack DMR1, together with the overall high resemblance of the mouse gene to the human SNRPN, offers an excellent experimental tool to study the regional control of this imprinted chromosomal domain.

Complexities in the genetic structure of Anopheles gambiae populations in west Africa as revealed by microsatellite DNA analysis

Chromosomal forms of Anopheles gambiae, given the informal designations Bamako, Mopti, and Savannah, have been recognized by the presence or absence of four paracentric inversions on chromosome 2. Studies of karyotype frequencies at sites where the forms occur in sympatry have led to the suggestion that these forms represent species. We conducted a study of the genetic structure of populations of An. gambiae from two villages in Mali, west Africa. Populations at each site were composed of the Bamako and Mopti forms and the sibling species, Anopheles arabiensis. Karyotypes were determined for each individual mosquito and genotypes at 21 microsatellite loci determined. A number of the microsatellites have been physically mapped to polytene chromosomes, making it possible to select loci based on their position relative to the inversions used to define forms. We found that the chromosomal forms differ at all loci on chromosome 2, but there were few differences for loci on other chromosomes. Geographic variation was small. Gene flow appears to vary among different regions within the genome, being lowest on chromosome 2, probably due to hitchhiking with the inversions. We conclude that the majority of observed genetic divergence between chromosomal forms can be explained by forces that need not involve reproductive isolation, although reproductive isolation is not ruled out. We found low levels of gene flow between the sibling species Anopheles gambiae and Anopheles arabiensis, similar to estimates based on observed frequencies of hybrid karyotypes in natural populations.

Major recent and independent changes in levels and patterns of expression have occurred at the b gene, a regulatory locus in maize

The b locus encodes a transcription factor that regulates the expression of genes that produce purple anthocyanin pigment. Different b alleles are expressed in distinct tissues, causing tissue-specific anthocyanin production. Understanding how phenotypic diversity is produced and maintained at the b locus should provide models for how other regulatory genes, including those that influence morphological traits and development, evolve. We have investigated how different levels and patterns of pigmentation have evolved by determining the phenotypic and evolutionary relationships between 18 alleles that represent the diversity of b alleles in Zea mays. Although most of these alleles have few phenotypic differences, five alleles have very distinct tissue-specific patterns of pigmentation. Superimposing the phenotypes on the molecular phylogeny reveals that the alleles with strong and distinctive patterns of expression are closely related to alleles with weak expression, implying that the distinctive patterns have arisen recently. We have identified apparent insertions in three of the five phenotypically distinct alleles, and the fourth has unique upstream restriction fragment length polymorphisms relative to closely related alleles. The insertion in B-Peru has been shown to be responsible for its unique expression and, in the other two alleles, the presence of the insertion correlates with the phenotype. These results suggest that major changes in gene expression are probably the result of large-scale changes in DNA sequence and/or structure most likely mediated by transposable elements.

Dissociation of Oct-1 from the Nuclear Peripheral Structure Induces the Cellular Aging-associated Collagenase Gene Expression

The cellular aging-associated transcriptional repressor that we previously named as Orpheus was identical to Oct-1, a member of the POU domain family. Oct-1 represses the collagenase gene, one of the cellular aging-associated genes, by interacting with an AT-rich cis-element in the upstream of the gene in preimmortalized cells at earlier population-doubling levels and in immortalized cells. In these stages of cells, considerable fractions of the Oct-1 protein were prominently localized in the nuclear periphery and colocalized with lamin B. During the cellular aging process, however, this subspecies of Oct-1 disappeared from the nuclear periphery. The cells lacking the nuclear peripheral Oct-1 protein exhibited strong collagenase expression and carried typical senescent morphologies. Concomitantly, the binding activity and the amount of nuclear Oct-1 protein were reduced in the aging process and resumed after immortalization. However, the whole cellular amounts of Oct-1 protein were not significantly changed during either process. Thus, the cellular aging-associated genes including the collagenase gene seemed to be derepressed by the dissociation of Oct-1 protein from the nuclear peripheral structure. Oct-1 may form a transcriptional repressive apparatus by anchoring nuclear matrix attachment regions onto the nuclear lamina in the nuclear periphery.

Substitutions in the aspartate transcarbamoylase domain of hamster CAD disrupt oligomeric structure

Aspartate transcarbamoylase (ATCase; EC 2.1.3.2) is one of three enzymatic domains of CAD, a protein whose native structure is usually a hexamer of identical subunits. Alanine substitutions for the ATCase residues Asp-90 and Arg-269 were generated in a bicistronic vector that encodes a 6-histidine-tagged hamster CAD. Stably transfected mammalian cells expressing high levels of CAD were easily isolated and CAD purification was simplified over previous procedures. The substitutions reduce the ATCase Vmax of the altered CADs by 11-fold and 46-fold, respectively, as well as affect the enzyme's affinity for aspartate. At 25 mM Mg2+, these substitutions cause the oligomeric CAD to dissociate into monomers. Under the same dissociating conditions, incubating the altered CAD with the ATCase substrate carbamoyl phosphate or the bisubstrate analogue N-phosphonacetyl-l-aspartate unexpectedly leads to the reformation of hexamers. Incubation with the other ATCase substrate, aspartate, has no effect. These results demonstrate that the ATCase domain is central to hexamer formation in CAD and suggest that the ATCase reaction mechanism is ordered in the same manner as the Escherichia coli ATCase. Finally, the data indicate that the binding of carbamoyl phosphate induces conformational changes that enhance the interaction of CAD subunits.

Relationship between the oxidation potential and electron spin density of the primary electron donor in reaction centers from Rhodobacter sphaeroides

The primary electron donor in bacterial reaction centers is a dimer of bacteriochlorophyll a molecules, labeled L or M based on their proximity to the symmetry-related protein subunits. The electronic structure of the bacteriochlorophyll dimer was probed by introducing small systematic variations in the bacteriochlorophyll–protein interactions by a series of site-directed mutations that replaced residue Leu M160 with histidine, tyrosine, glutamic acid, glutamine, aspartic acid, asparagine, lysine, and serine. The midpoint potentials for oxidation of the dimer in the mutants showed an almost continuous increase up to ≈60 mV compared with wild type. The spin density distribution of the unpaired electron in the cation radical state of the dimer was determined by electron–nuclear–nuclear triple resonance spectroscopy in solution. The ratio of the spin density on the L side of the dimer to the M side varied from ≈2:1 to ≈5:1 in the mutants compared with ≈2:1 for wild type. The correlation between the midpoint potential and spin density distribution was described using a simple molecular orbital model, in which the major effect of the mutations is assumed to be a change in the energy of the M half of the dimer, providing estimates for the coupling and energy levels of the orbitals in the dimer. These results demonstrate that the midpoint potential can be fine-tuned by electrostatic interactions with amino acids near the dimer and show that the properties of the electronic structure of a donor or acceptor in a protein complex can be directly related to functional properties such as the oxidation–reduction midpoint potential.

Endogenous Msx1 antisense transcript: In vivo and in vitro evidences, structure, and potential involvement in skeleton development in mammals

Msx1 is a key factor for the development of tooth and craniofacial skeleton and has been proposed to play a pivotal role in terminal cell differentiation. In this paper, we demonstrated the presence of an endogenous Msx1 antisense RNA (Msx1-AS RNA) in mice, rats, and humans. In situ analysis revealed that this RNA is expressed only in differentiated dental and bone cells with an inverse correlation with Msx1 protein. These in vivo data and overexpression of Msx1 sense and AS RNA in an odontoblastic cell line (MO6-G3) showed that the balance between the levels of the two Msx1 RNAs is related to the expression of Msx1 protein. To analyze the impact of this balance in the Msx-Dlx homeoprotein pathway, we analyzed the effect of Msx1, Msx2, and Dlx5 overexpression on proteins involved in skeletal differentiation. We showed that the Msx1-AS RNA is involved in crosstalk between the Msx-Dlx pathways because its expression was abolished by Dlx5. Msx1 was shown to down-regulate a master gene of skeletal cells differentiation, Cbfa1. All these data strongly suggest that the ratio between Msx1 sense and antisense RNAs is a very important factor in the control of skeletal terminal differentiation. Finally, the initiation site for Msx1-AS RNA transcription was located by primer extension in both mouse and human in an identical region, including a consensus TATA box, suggesting an evolutionary conservation of the AS RNA-mediated regulation of Msx1 gene expression.

In Vivo Ubiquinone Reduction Levels during Thermogenesis in Araceae1

In vivo ubiquinone (UQ) reduction levels were measured during the development of the inflorescences of Arum maculatum and Amorphophallus krausei. Thermogenesis in A. maculatum spadices appeared not to be confined to a single developmental stage, but occurred during various stages. The UQ pool in both A. maculatum and A. krausei appendices was approximately 90% reduced during thermogenesis. Respiratory characteristics of isolated appendix mitochondria did not change in the period around thermogenesis. Apparently, synthesis of the required enzyme capacity is regulated via a coarse control upon which a fine control of metabolism that regulates the onset of thermogenesis is imposed.

Structure and Expression of a Dhurrinase (β-Glucosidase) from Sorghum1

Sorghum (Sorghum bicolor L. Moench) has two isozymes of the cyanogenic β-glucosidase dhurrinase: dhurrinase-1 (Dhr1) and dhurrinase-2 (Dhr2). A nearly full-length cDNA encoding dhurrinase was isolated from 4-d-old etiolated seedlings and sequenced. The cDNA has a 1695-nucleotide-long open reading frame, which codes for a 565-amino acid-long precursor and a 514-amino acid-long mature protein, respectively. Deduced amino acid sequence of the sorghum Dhr showed 70% identity with two maize (Zea mays) β-glucosidase isozymes. Southern-blot data suggested that β-glu-cosidase is encoded by a small multigene family in sorghum. Northern-blot data indicated that the mRNA corresponding to the cloned Dhr cDNA is present at high levels in the node and upper half of the mesocotyl in etiolated seedlings but at low levels in the root—only in the zone of elongation and the tip region. Light-grown seedling parts had lower levels of Dhr mRNA than those of etiolated seedlings. Immunoblot analysis performed using maize-anti-β-glucosidase sera detected two distinct dhurrinases (57 and 62 kD) in sorghum. The distribution of Dhr activity in different plant parts supports the mRNA and immunoreactive protein data, suggesting that the cloned cDNA corresponds to the Dhr1 (57 kD) isozyme and that the dhr1 gene shows organ-specific expression.

Two Isoforms of NADPH:Cytochrome P450 Reductase in Arabidopsis thaliana : Gene Structure, Heterologous Expression in Insect Cells, and Differential Regulation

We have investigated two NADPH-cytochrome (Cyt) P450 reductase isoforms encoded by separate genes (AR1 and AR2) in Arabidopsis thaliana. We isolated AR1 and AR2 cDNAs using a mung bean (Phaseolus aureus L.) NADPH-Cyt P450 reductase cDNA as a probe. The recombinant AR1 and AR2 proteins produced using a baculovirus expression system showed similar Km values for Cyt c and NADPH, respectively. In the reconstitution system with a recombinant cinnamate 4-hydroxylase (CYP73A5), the recombinant AR1 and AR2 proteins gave the same level of cinnamate 4-hydroxylase activity (about 70 nmol min−1 nmol−1 P450). The AR2 gene expression was transiently induced by 4- and 3-fold within 1 h of wounding and light treatments, respectively, and the induction time course preceded those of CYP73A5 and a phenylalanine ammonia-lyase (PAL1) gene. On the contrary, the AR1 expression level did not change during the treatments. Analysis of the AR1 and AR2 gene structure revealed that only the AR2 promoter contained three putative sequence motifs (boxes P, A, and L), which are involved in the coordinated expression of CYP73A5 and other phenylpropanoid pathway genes. These results suggest the possibility that AR2 transcription may be functionally linked to the induced levels of phenylpropanoid pathway enzymes.