Life cycle assessment as a policy-support tool: the case of taxis in the city of Madrid

This paper examined the potentialities of Life Cycle Assessment (LCA) as instrument for policy-support. To this respect, the adoption of an initiative within the Madrid Air Quality Plan (AQP) 2011–2015 regarding the substitution of diesel taxis with hybrid, natural gas and LPG alternatives was studied. Four different scenarios were elaborated, a business-as-usual scenario (BAU), the scenario of the AQP, and two extreme-situation scenarios: all-diesel (ADI) and all-ecologic (AEC). Impacts were characterized according to the ILCD methodology, focusing especially on climate change (CC) and photochemical ozone formation (PO). SimaPro 7.3 was used as analysis and inventory-construction tool. The results indicate that the shift to ecologic alternatives reduced impacts, especially those related to CC and PO. For the complete life cycle, reductions of 13% (CC) and 25% (PO) were observed for AQP against BAU (CC:1365 GgCO2, PO:13336 MgNMVOC). Deeper reductions were observed for AEC (CC:34%, PO:59%), while ADI produced slight increases in impacts if against BAU. The analysis of the use-phase revealed that the central and highest speed zones of the city benefit from the adoption of AQP. This is especially evident in zone 7, with reductions of 16% in CC and 31% in PO respectively against BAU (CCzone1:3443 kgCO2/veh·km, POzone7:11.1 kgNMVOC/veh·km).

Development of the infrastructure for a multi-agent traffic management system simulation

This document contains detailed description of the design and the implementation of a multi-agent application controlling traffic lights in a city together with a system for simulating traffic and testing. The goal of this thesis is to design and build a simplified intelligent and distributed solution to the problem with the traffic in the big cities following different good practices in order to allow future refining of the model of the real world. The problem of the traffic in the big cities is still a problem that cannot be solved. Not only is the increasing number of cars a reason for the traffic jams, but also the way the traffic is organized. Usually, the intersections with traffic lights are replaced by roundabouts or interchanges to increase the number of cars that can cross the intersection in certain time. But still there are places where the infrastructure cannot be changed and the traffic light semaphores are the only way to control the car flows. In real life, the traffic lights have a predefined plan for change or they receive information from a centralized system when and how they have to change. But what if the traffic lights can cooperate and decide on their own when and how to change? Using this problem, the purpose of the thesis is to explore different agent-based software engineering approaches to design and build a non-conventional distributed system. From the software engineering point of view, the goal of the thesis is to apply the knowledge and use the skills, acquired during the various courses of the master program in Software Engineering, while solving a practical and complex problem such as the traffic in the cities.

A system of systems engineering approach to design new smarter and energy efficient products

The concept of smartness of energy efficient products and systems from a business perspective has been investigated by several authors. The problem of understanding, designing, engineering and governing these technologies requires new concepts. The emergence of these modern technologies causes a myriad of interconnected systems, which are working together to satisfy the necessities of modern life. The problem of understanding, designing, engineering, and governing these technologies requires new concepts. Development of System of System Engineering (SoSE) is an attempt by the systems engineering and science community to fulfill this requirement.

Aplicación de la técnica de inyección del terreno por fracturación hidráulica a las cimentaciones de puentes en circunstancias de especial dificultad

La implantación de una gran obra de paso facilita en multitud de ocasiones la vida de aquellos que aprovechen las ventajas que ofrece esta estructura. Sin embargo, el camino que lleva a su construcción supone importantes y tortuosos retos cuando el terreno de cimentación presenta condiciones geotécnicas no compatibles con las grandes cargas que el viaducto necesita transmitirle para su buen funcionamiento. El trabajo que desarrolla esta Tesis Doctoral proporciona una herramienta eficaz y económica, por lo reducido de su extensión y medios, que permite allanar el camino que acomete la construcción de una estructura de tal envergadura. Mediante el análisis de la problemática del terreno y de las distintas soluciones de cimentación empleadas en la actualidad, se conduce al lector hacia una técnica de cimentación innovadora que combina la técnica del micropilotaje, para la canalización de las cargas estructurales, junto a la técnica de la inyección de fracturación hidráulica, que mejorará el terreno de implantación de los micropilotes, conformando así un medio capaz de recibir y transmitir grandes cargas en cualquier tipo de terreno. La técnica ya empleada del micropilotaje, por su trabajo esencialmente axil, requiere, aunque sin problema, la constitución de sistemas de fuerzas que equilibren el sistema de cargas provenientes del viaducto. Pero su capacidad resistente viene condicionada por el terreno circundante. Cuando la roca rodea el micropilote, su empleo se realiza sin problema y sin necesidad de mejorar el terreno. Pero sin terreno consistente, el empleo de inyecciones de fracturación hidráulica a través de los propios micropilotes, no sólo mejorará la capacidad resistente de los micropilotes, aumentando la inercia necesaria cuando las cargas son de origen sísmico o estructural ferroviario, sino que resolverá y eliminará los problemas de estabilidad que presentan las laderas que frecuentemente deben recibir las cargas de la obra de paso. Tras recoger el análisis ya realizado en el Trabajo de Investigación, donde se justificaba la alta capacidad resistente de un micropilote con terreno circundante mejorado por la inyección, en la actual Tesis se emplean modelos matemáticos sobre un caso real de viaducto sometido a un gran sismo e implantado en una enorme quebrada (500 m) de Colombia, sujeta a lluvias torrenciales y en la zona de mayor sismicidad del país. Con ello se comprueba la estabilización que se alcanza en el terreno de cimentación con el empleo de esta técnica de transmisión de carga y mejora del terreno. De esta forma se completa un ciclo que justifica las bondades de esta combinación de técnicas de cimentación, pero se abren las puertas a nuevos entornos de aplicación, como edificios antiguos de cualquier tipo que requieran recalces, y no sólo en la implantación de grandes obras de paso. ABSTRACT The establishment of a large bridge represents, in many cases, a better life for those who can take profit of the advantages provided by that structure. Nevertheless, the process of building this structure has to overcome important and difficult circumstances whenever the geotechnical conditions of the bridge site are not adequate to carry the large loads transmitted by the bridge structure. This study develops a method both effective and economical, due to the extension and means necessary for its application, which allows to solve properly the foundation of a structure of that importance. Considering the geotechnical problems inherent to the bridge site, along with the different foundation solutions that are presently used and their limitations, the study leads the reader to an innovative technique which combines the micropile system, for transmission of the structural loads of the bridge, with the technique of hydraulic fracture grouting for improvement of the ground around the micropiles, allowing to both stabilizing and transmitting large loads in any kind of ground. It is well known that the micropiles work axially, and this condition requires an adequate distribution of those units, in order to properly absorb the load system introduced by the viaduct. The resistance of the bridge foundation is, in any case, provided by the ground. When rock is encountered, the micropiles have been successfully used without improving the ground. However, as it is shown in this study, by using the micropiles as sleeve-pipes for hydraulic fracture grouting, not only the micropile resistance can be improved in any ground, but it is possible to develop grouted “solids” in the ground, whose inertia allows to absorb actions of structural and seismic origin. Additionally, as it is shown and analyzed in the Thesis, the ground improvement can give an adequate safety factor to the slopes frequently encountered in bridge sites. In order to properly justify those advantages of combining micropiles with ground improvement through fracture grouting around the micropiles, mathematical models have been developed and applied to a real case of a cable-stayed bridge installed on a very large ravine (500 m) in Colombia, located in the highest seismic zone of the country, and subject to torrential rains. The results of this numerical analysis show the high safety condition provided by the ground improvement to the viaduct site. In conclusion, the Thesis shows the important improvement that can be provided by the combination of micropiles and soil improvement, through fracture grouting, to the problem of founding bridges. However, it can be understood that this technique could be applied successfully to underpinning buildings, specially old buildings of any type, apart from its use in bridge foundations.

Development and implementation of an air quality integrated assessment modelling system for the Iberian Peninsula

La mejora de la calidad del aire es una tarea eminentemente interdisciplinaria. Dada la gran variedad de ciencias y partes involucradas, dicha mejora requiere de herramientas de evaluación simples y completamente integradas. La modelización para la evaluación integrada (integrated assessment modeling) ha demostrado ser una solución adecuada para la descripción de los sistemas de contaminación atmosférica puesto que considera cada una de las etapas involucradas: emisiones, química y dispersión atmosférica, impactos ambientales asociados y potencial de disminución. Varios modelos de evaluación integrada ya están disponibles a escala continental, cubriendo cada una de las etapas antesmencionadas, siendo el modelo GAINS (Greenhouse Gas and Air Pollution Interactions and Synergies) el más reconocido y usado en el contexto europeo de toma de decisiones medioambientales. Sin embargo, el manejo de la calidad del aire a escala nacional/regional dentro del marco de la evaluación integrada es deseable. Esto sin embargo, no se lleva a cabo de manera satisfactoria con modelos a escala europea debido a la falta de resolución espacial o de detalle en los datos auxiliares, principalmente los inventarios de emisión y los patrones meteorológicos, entre otros. El objetivo de esta tesis es presentar los desarrollos en el diseño y aplicación de un modelo de evaluación integrada especialmente concebido para España y Portugal. El modelo AERIS (Atmospheric Evaluation and Research Integrated system for Spain) es capaz de cuantificar perfiles de concentración para varios contaminantes (NO2, SO2, PM10, PM2,5, NH3 y O3), el depósito atmosférico de especies de azufre y nitrógeno así como sus impactos en cultivos, vegetación, ecosistemas y salud como respuesta a cambios porcentuales en las emisiones de sectores relevantes. La versión actual de AERIS considera 20 sectores de emisión, ya sea equivalentes a sectores individuales SNAP o macrosectores, cuya contribución a los niveles de calidad del aire, depósito e impactos han sido modelados a través de matrices fuentereceptor (SRMs). Estas matrices son constantes de proporcionalidad que relacionan cambios en emisiones con diferentes indicadores de calidad del aire y han sido obtenidas a través de parametrizaciones estadísticas de un modelo de calidad del aire (AQM). Para el caso concreto de AERIS, su modelo de calidad del aire “de origen” consistió en el modelo WRF para la meteorología y en el modelo CMAQ para los procesos químico-atmosféricos. La cuantificación del depósito atmosférico, de los impactos en ecosistemas, cultivos, vegetación y salud humana se ha realizado siguiendo las metodologías estándar establecidas bajo los marcos internacionales de negociación, tales como CLRTAP. La estructura de programación está basada en MATLAB®, permitiendo gran compatibilidad con software típico de escritorio comoMicrosoft Excel® o ArcGIS®. En relación con los niveles de calidad del aire, AERIS es capaz de proveer datos de media anual y media mensual, así como el 19o valor horario más alto paraNO2, el 25o valor horario y el 4o valor diario más altos para SO2, el 36o valor diario más alto para PM10, el 26o valor octohorario más alto, SOMO35 y AOT40 para O3. En relación al depósito atmosférico, el depósito acumulado anual por unidad de area de especies de nitrógeno oxidado y reducido al igual que de azufre pueden ser determinados. Cuando los valores anteriormente mencionados se relacionan con características del dominio modelado tales como uso de suelo, cubiertas vegetales y forestales, censos poblacionales o estudios epidemiológicos, un gran número de impactos puede ser calculado. Centrándose en los impactos a ecosistemas y suelos, AERIS es capaz de estimar las superaciones de cargas críticas y las superaciones medias acumuladas para especies de nitrógeno y azufre. Los daños a bosques se calculan como una superación de los niveles críticos de NO2 y SO2 establecidos. Además, AERIS es capaz de cuantificar daños causados por O3 y SO2 en vid, maíz, patata, arroz, girasol, tabaco, tomate, sandía y trigo. Los impactos en salud humana han sido modelados como consecuencia de la exposición a PM2,5 y O3 y cuantificados como pérdidas en la esperanza de vida estadística e indicadores de mortalidad prematura. La exactitud del modelo de evaluación integrada ha sido contrastada estadísticamente con los resultados obtenidos por el modelo de calidad del aire convencional, exhibiendo en la mayoría de los casos un buen nivel de correspondencia. Debido a que la cuantificación de los impactos no es llevada a cabo directamente por el modelo de calidad del aire, un análisis de credibilidad ha sido realizado mediante la comparación de los resultados de AERIS con los de GAINS para un escenario de emisiones determinado. El análisis reveló un buen nivel de correspondencia en las medias y en las distribuciones probabilísticas de los conjuntos de datos. Las pruebas de verificación que fueron aplicadas a AERIS sugieren que los resultados son suficientemente consistentes para ser considerados como razonables y realistas. En conclusión, la principal motivación para la creación del modelo fue el producir una herramienta confiable y a la vez simple para el soporte de las partes involucradas en la toma de decisiones, de cara a analizar diferentes escenarios “y si” con un bajo coste computacional. La interacción con políticos y otros actores dictó encontrar un compromiso entre la complejidad del modeladomedioambiental con el carácter conciso de las políticas, siendo esto algo que AERIS refleja en sus estructuras conceptual y computacional. Finalmente, cabe decir que AERIS ha sido creado para su uso exclusivo dentro de un marco de evaluación y de ninguna manera debe ser considerado como un sustituto de los modelos de calidad del aire ordinarios. ABSTRACT Improving air quality is an eminently inter-disciplinary task. The wide variety of sciences and stakeholders that are involved call for having simple yet fully-integrated and reliable evaluation tools available. Integrated AssessmentModeling has proved to be a suitable solution for the description of air pollution systems due to the fact that it considers each of the involved stages: emissions, atmospheric chemistry, dispersion, environmental impacts and abatement potentials. Some integrated assessment models are available at European scale that cover each of the before mentioned stages, being the Greenhouse Gas and Air Pollution Interactions and Synergies (GAINS) model the most recognized and widely-used within a European policy-making context. However, addressing air quality at the national/regional scale under an integrated assessment framework is desirable. To do so, European-scale models do not provide enough spatial resolution or detail in their ancillary data sources, mainly emission inventories and local meteorology patterns as well as associated results. The objective of this dissertation is to present the developments in the design and application of an Integrated Assessment Model especially conceived for Spain and Portugal. The Atmospheric Evaluation and Research Integrated system for Spain (AERIS) is able to quantify concentration profiles for several pollutants (NO2, SO2, PM10, PM2.5, NH3 and O3), the atmospheric deposition of sulfur and nitrogen species and their related impacts on crops, vegetation, ecosystems and health as a response to percentual changes in the emissions of relevant sectors. The current version of AERIS considers 20 emission sectors, either corresponding to individual SNAP sectors or macrosectors, whose contribution to air quality levels, deposition and impacts have been modeled through the use of source-receptor matrices (SRMs). Thesematrices are proportionality constants that relate emission changes with different air quality indicators and have been derived through statistical parameterizations of an air qualitymodeling system (AQM). For the concrete case of AERIS, its parent AQM relied on the WRF model for meteorology and on the CMAQ model for atmospheric chemical processes. The quantification of atmospheric deposition, impacts on ecosystems, crops, vegetation and human health has been carried out following the standard methodologies established under international negotiation frameworks such as CLRTAP. The programming structure isMATLAB ® -based, allowing great compatibility with typical software such as Microsoft Excel ® or ArcGIS ® Regarding air quality levels, AERIS is able to provide mean annual andmean monthly concentration values, as well as the indicators established in Directive 2008/50/EC, namely the 19th highest hourly value for NO2, the 25th highest daily value and the 4th highest hourly value for SO2, the 36th highest daily value of PM10, the 26th highest maximum 8-hour daily value, SOMO35 and AOT40 for O3. Regarding atmospheric deposition, the annual accumulated deposition per unit of area of species of oxidized and reduced nitrogen as well as sulfur can be estimated. When relating the before mentioned values with specific characteristics of the modeling domain such as land use, forest and crops covers, population counts and epidemiological studies, a wide array of impacts can be calculated. When focusing on impacts on ecosystems and soils, AERIS is able to estimate critical load exceedances and accumulated average exceedances for nitrogen and sulfur species. Damage on forests is estimated as an exceedance of established critical levels of NO2 and SO2. Additionally, AERIS is able to quantify damage caused by O3 and SO2 on grapes, maize, potato, rice, sunflower, tobacco, tomato, watermelon and wheat. Impacts on human health aremodeled as a consequence of exposure to PM2.5 and O3 and quantified as losses in statistical life expectancy and premature mortality indicators. The accuracy of the IAM has been tested by statistically contrasting the obtained results with those yielded by the conventional AQM, exhibiting in most cases a good agreement level. Due to the fact that impacts cannot be directly produced by the AQM, a credibility analysis was carried out for the outputs of AERIS for a given emission scenario by comparing them through probability tests against the performance of GAINS for the same scenario. This analysis revealed a good correspondence in the mean behavior and the probabilistic distributions of the datasets. The verification tests that were applied to AERIS suggest that results are consistent enough to be credited as reasonable and realistic. In conclusion, the main reason thatmotivated the creation of this model was to produce a reliable yet simple screening tool that would provide decision and policy making support for different “what-if” scenarios at a low computing cost. The interaction with politicians and other stakeholders dictated that reconciling the complexity of modeling with the conciseness of policies should be reflected by AERIS in both, its conceptual and computational structures. It should be noted however, that AERIS has been created under a policy-driven framework and by no means should be considered as a substitute of the ordinary AQM.

Métodos diferenciales e interferométricos para la evaluación de deformaciones de la corteza terrestre mediante técnicas GNSS e InSAR = Differential and interferometric methods for evaluation of crustal deformation using GNSS and InSAR techniques

La actividad volcánica interviene en multitud de facetas de la propia actividad humana, no siempre negativas. Sin embargo, son más los motivos de peligrosidad y riesgo que incitan al estudio de la actividad volcánica. Existen razones de seguridad que inciden en el mantenimiento del seguimiento y monitorización de la actividad volcánica para garantizar la vida y la seguridad de los asentamientos antrópicos en las proximidades de los edificios volcánicos. En esta tesis se define e implementa un sistema de monitorización de movimientos de la corteza en las islas de Tenerife y La Palma, donde el impacto social que representa un aumento o variación de la actividad volcánica en las islas es muy severo. Aparte de la alta densidad demográfica del Archipiélago, esta población aumenta significativamente, en diferentes periodos a lo largo del año, debido a la actividad turística que representa la mayor fuente de ingresos de las islas. La población y los centros turísticos se diseminan predominantemente a lo largo de las costas y también a lo largo de los flancos de los edificios volcánicos. Quizá el mantenimiento de estas estructuras sociales y socio-económicas son los motivos más importantes que justifican una monitorización de la actividad volcánica en las Islas Canarias. Recientemente se ha venido trabajando cada vez más en el intento de predecir la actividad volcánica utilizando los nuevos sistemas de monitorización geodésica, puesto que la actividad volcánica se manifiesta anteriormente por deformación de la corteza terrestre y cambios en la fuerza de la gravedad en la zona donde más tarde se registran eventos volcánicos. Los nuevos dispositivos y sensores que se han desarrollado en los últimos años en materias como la geodesia, la observación de la Tierra desde el espacio y el posicionamiento por satélite, han permitido observar y medir tanto la deformación producida en el terreno como los cambios de la fuerza de la gravedad antes, durante y posteriormente a los eventos volcánicos que se producen. Estos nuevos dispositivos y sensores han cambiado las técnicas o metodologías geodésicas que se venían utilizando hasta la aparición de los mismos, renovando métodos clásicos y desarrollando otros nuevos que ya se están afianzando como metodologías probadas y reconocidas para ser usadas en la monitorización volcánica. Desde finales de la década de los noventa del siglo pasado se han venido desarrollando en las Islas Canarias varios proyectos que han tenido como objetivos principales el desarrollo de nuevas técnicas de observación y monitorización por un lado y el diseño de una metodología de monitorización volcánica adecuada, por otro. Se presenta aquí el estudio y desarrollo de técnicas GNSS para la monitorización de deformaciones corticales y su campo de velocidades para las islas de Tenerife y La Palma. En su implementación, se ha tenido en cuenta el uso de la infraestructura geodésica y de monitorización existente en el archipiélago a fin de optimizar costes, además de complementarla con nuevas estaciones para dar una cobertura total a las dos islas. Los resultados obtenidos en los proyectos, que se describen en esta memoria, han dado nuevas perspectivas en la monitorización geodésica de la actividad volcánica y nuevas zonas de interés que anteriormente no se conocían en el entorno de las Islas Canarias. Se ha tenido especial cuidado en el tratamiento y propagación de los errores durante todo el proceso de observación, medida y proceso de los datos registrados, todo ello en aras de cuantificar el grado de fiabilidad de los resultados obtenidos. También en este sentido, los resultados obtenidos han sido verificados con otros procedentes de sistemas de observación radar de satélite, incorporando además a este estudio las implicaciones que el uso conjunto de tecnologías radar y GNSS tendrán en un futuro en la monitorización de deformaciones de la corteza terrestre. ABSTRACT Volcanic activity occurs in many aspects of human activity, and not always in a negative manner. Nonetheless, research into volcanic activity is more likely to be motivated by its danger and risk. There are security reasons that influence the monitoring of volcanic activity in order to guarantee the life and safety of human settlements near volcanic edifices. This thesis defines and implements a monitoring system of movements in the Earth’s crust in the islands of Tenerife and La Palma, where the social impact of an increase (or variation) of volcanic activity is very severe. Aside from the high demographic density of the archipelago, the population increases significantly in different periods throughout the year due to tourism, which represents a major source of revenue for the islands. The population and the tourist centres are mainly spread along the coasts and also along the flanks of the volcanic edifices. Perhaps the preservation of these social and socio-economic structures is the most important reason that justifies monitoring volcanic activity in the Canary Islands. Recently more and more work has been done with the intention of predicting volcanic activity, using new geodesic monitoring systems, since volcanic activity is evident prior to eruption because of a deformation of the Earth’s crust and changes in the force of gravity in the zone where volcanic events will later be recorded. The new devices and sensors that have been developed in recent years in areas such as geodesy, the observation of the Earth from space, and satellite positioning have allowed us to observe and measure the deformation produced in the Earth as well as the changes in the force of gravity before, during, and after the volcanic events occur. The new devices and sensors have changed the geodetic techniques and methodologies that were used previously. The classic methods have been renovated and other newer ones developed that are now vouched for as proven recognised methodologies to be used for volcanic monitoring. Since the end of the 1990s, in the Canary Islands various projects have been developed whose principal aim has been the development of new observation and monitoring techniques on the one hand, and the design of an appropriate volcanic monitoring methodology on the other. The study and development of GNSS techniques for the monitoring of crustal deformations and their velocity field is presented here. To carry out the study, the use of geodetic infrastructure and existing monitoring in the archipelago have been taken into account in order to optimise costs, besides complementing it with new stations for total coverage on both islands. The results obtained in the projects, which are described below, have produced new perspectives in the geodetic monitoring of volcanic activity and new zones of interest which previously were unknown in the environment of the Canary Islands. Special care has been taken with the treatment and propagation of errors during the entire process of observing, measuring, and processing the recorded data. All of this was done in order to quantify the degree of trustworthiness of the results obtained. Also in this sense, the results obtained have been verified with others from satellite radar observation systems, incorporating as well in this study the implications that the joint use of radar technologies and GNSS will have for the future of monitoring deformations in the Earth’s crust.

Ol-Prx 3, a member of an additional class of homeobox genes, is unimodally expressed in several domains of the developing and adult central nervous system of the medaka (Oryzias latipes)

Large-scale genetic screens for mutations affecting early neurogenesis of vertebrates have recently been performed with an aquarium fish, the zebrafish. Later stages of neural morphogenesis have attracted less attention in small fish species, partly because of the lack of molecular markers of developing structures that may facilitate the detection of discrete structural alterations. In this context, we report the characterization of Ol-Prx 3 (Oryzias latipes-Prx 3). This gene was isolated in the course of a large-scale screen for brain cDNAs containing a highly conserved DNA binding region, the homeobox helix-three. Sequence analysis revealed that this gene belongs to another class of homeobox genes, together with a previously isolated mouse ortholog, called OG-12 [Rovescalli, A. C., Asoh, S. & Nirenberg, M. (1996) Proc. Natl. Acad. Sci. USA 93, 10691–10696] and with the human SHOX gene [Rao, E., Weiss, B., Fukami, M., Rump, A., Niesler, B., et al. (1997) Nat. Genet. 16, 54–62], thought to be involved in the short-stature phenotype of Turner syndrome patients. These three genes exhibit a moderate level of identity in the homeobox with the other genes of the paired-related (PRX) gene family. Ol-Prx 3, as well as the PRX genes, are expressed in various cartilaginous structures of head and limbs. These genes might thus be involved in common regulatory pathways during the morphogenesis of these structures. Moreover, this paper reports a complex and monophasic pattern of Ol-Prx 3 expression in the central nervous system, which differs markedly from the patterns reported for the PRX genes, Prx 3 excluded: this gene begins to be expressed in a variety of central nervous system territories at late neurula stage. Strikingly, it remains turned on in some of the derivatives of each territory during the entire life of the fish. We hope this work will thus help identify common features for the PRX 3 family of homeobox genes.

Postnatal mouse subventricular zone neuronal precursors can migrate and differentiate within multiple levels of the developing neuraxis

The mammalian subventricular zone (SVZ) of the lateral wall of the forebrain ventricle retains a population of proliferating neuronal precursors throughout life. Neuronal precursors born in the postnatal and adult SVZ migrate to the olfactory bulb where they differentiate into interneurons. Here we tested the potential of mouse postnatal SVZ precursors in the environment of the embryonic brain: (i) a ubiquitous genetic marker, (ii) a neuron-specific transgene, and (iii) a lipophilic-dye were used to follow the fate of postnatal day 5–10 SVZ cells grafted into embryonic mouse brain ventricles at day 15 of gestation. Graft-derived cells were found at multiple levels of the neuraxis, including septum, thalamus, hypothalamus, and in large numbers in the midbrain inferior colliculus. We observed no integration into the cortex. Neuronal differentiation of graft derived cells was demonstrated by double-staining with neuron-specific β-tubulin antibodies, expression of the neuron-specific transgene, and the dendritic arbors revealed by the lipophilic dye. We conclude that postnatal SVZ cells can migrate through and differentiate into neurons within multiple embryonic brain regions other than the olfactory bulb.

Delayed symptom onset and increased life expectancy in Sandhoff disease mice treated with N-butyldeoxynojirimycin

Sandhoff disease is a neurodegenerative disorder resulting from the autosomal recessive inheritance of mutations in the HEXB gene, which encodes the β-subunit of β-hexosaminidase. GM2 ganglioside fails to be degraded and accumulates within lysosomes in cells of the periphery and the central nervous system (CNS). There are currently no therapies for the glycosphingolipid lysosomal storage diseases that involve CNS pathology, including the GM2 gangliosidoses. One strategy for treating this and related diseases is substrate deprivation. This would utilize an inhibitor of glycosphingolipid biosynthesis to balance synthesis with the impaired rate of catabolism, thus preventing storage. One such inhibitor is N-butyldeoxynojirimycin, which currently is in clinical trials for the potential treatment of type 1 Gaucher disease, a related disease that involves glycosphingolipid storage in peripheral tissues, but not in the CNS. In this study, we have evaluated whether this drug also could be applied to the treatment of diseases with CNS storage and pathology. We therefore have treated a mouse model of Sandhoff disease with the inhibitor N-butyldeoxynojirimycin. The treated mice have delayed symptom onset, reduced storage in the brain and peripheral tissues, and increased life expectancy. Substrate deprivation therefore offers a potentially general therapy for this family of lysosomal storage diseases, including those with CNS disease.

Biochemical evolution II: Origin of life in tubular microstructures on weathered feldspar surfaces

Mineral surfaces were important during the emergence of life on Earth because the assembly of the necessary complex biomolecules by random collisions in dilute aqueous solutions is implausible. Most silicate mineral surfaces are hydrophilic and organophobic and unsuitable for catalytic reactions, but some silica-rich surfaces of partly dealuminated feldspars and zeolites are organophilic and potentially catalytic. Weathered alkali feldspar crystals from granitic rocks at Shap, north west England, contain abundant tubular etch pits, typically 0.4–0.6 μm wide, forming an orthogonal honeycomb network in a surface zone 50 μm thick, with 2–3 × 106 intersections per mm2 of crystal surface. Surviving metamorphic rocks demonstrate that granites and acidic surface water were present on the Earth’s surface by ∼3.8 Ga. By analogy with Shap granite, honeycombed feldspar has considerable potential as a natural catalytic surface for the start of biochemical evolution. Biomolecules should have become available by catalysis of amino acids, etc. The honeycomb would have provided access to various mineral inclusions in the feldspar, particularly apatite and oxides, which contain phosphorus and transition metals necessary for energetic life. The organized environment would have protected complex molecules from dispersion into dilute solutions, from hydrolysis, and from UV radiation. Sub-micrometer tubes in the honeycomb might have acted as rudimentary cell walls for proto-organisms, which ultimately evolved a lipid lid giving further shelter from the hostile outside environment. A lid would finally have become a complete cell wall permitting detachment and flotation in primordial “soup.” Etch features on weathered alkali feldspar from Shap match the shape of overlying soil bacteria.

Oxygen isotope ratios of PO4: An inorganic indicator of enzymatic activity and P metabolism and a new biomarker in the search for life

The distinctive relations between biological activity and isotopic effect recorded in biomarkers (e.g., carbon and sulfur isotope ratios) have allowed scientists to suggest that life originated on this planet nearly 3.8 billion years ago. The existence of life on other planets may be similarly identified by geochemical biomarkers, including the oxygen isotope ratio of phosphate (δ18Op) presented here. At low near-surface temperatures, the exchange of oxygen isotopes between phosphate and water requires enzymatic catalysis. Because enzymes are indicative of cellular activity, the demonstration of enzyme-catalyzed PO4–H2O exchange is indicative of the presence of life. Results of laboratory experiments are presented that clearly show that δ18OP values of inorganic phosphate can be used to detect enzymatic activity and microbial metabolism of phosphate. Applications of δ18Op as a biomarker are presented for two Earth environments relevant to the search for extraterrestrial life: a shallow groundwater reservoir and a marine hydrothermal vent system. With the development of in situ analytical techniques and future planned sample return strategies, δ18Op may provide an important biosignature of the presence of life in extraterrestrial systems such as that on Mars.

Functional disorders of the sympathetic nervous system in mice lacking the α1B subunit (Cav 2.2) of N-type calcium channels

N-type voltage-dependent Ca2+ channels (VDCCs), predominantly localized in the nervous system, have been considered to play an essential role in a variety of neuronal functions, including neurotransmitter release at sympathetic nerve terminals. As a direct approach to elucidating the physiological significance of N-type VDCCs, we have generated mice genetically deficient in the α1B subunit (Cav 2.2). The α1B-deficient null mice, surprisingly, have a normal life span and are free from apparent behavioral defects. A complete and selective elimination of N-type currents, sensitive to ω-conotoxin GVIA, was observed without significant changes in the activity of other VDCC types in neuronal preparations of mutant mice. The baroreflex response, mediated by the sympathetic nervous system, was markedly reduced after bilateral carotid occlusion. In isolated left atria prepared from N-type-deficient mice, the positive inotropic responses to electrical sympathetic neuronal stimulation were dramatically decreased compared with those of normal mice. In contrast, parasympathetic nervous activity in the mutant mice was nearly identical to that of wild-type mice. Interestingly, the mutant mice showed sustained elevation of heart rate and blood pressure. These results provide direct evidence that N-type VDCCs are indispensable for the function of the sympathetic nervous system in circulatory regulation and indicate that N-type VDCC-deficient mice will be a useful model for studying disorders attributable to sympathetic nerve dysfunction.

Effects of Sulfanilamide and Methotrexate on 13C Fluxes through the Glycine Decarboxylase/Serine Hydroxymethyltransferase Enzyme System in Arabidopsis1

In C3 plants large amounts of photorespiratory glycine (Gly) are converted to serine by the tetrahydrofolate (THF)-dependent activities of the Gly decarboxylase complex (GDC) and serine hydroxymethyltransferase (SHMT). Using 13C nuclear magnetic resonance, we monitored the flux of carbon through the GDC/SHMT enzyme system in Arabidopsis thaliana (L.) Heynh. Columbia exposed to inhibitors of THF-synthesizing enzymes. Plants exposed for 96 h to sulfanilamide, a dihydropteroate synthase inhibitor, showed little reduction in flux through GDC/SHMT. Two other sulfonamide analogs were tested with similar results, although all three analogs competitively inhibited the partially purified enzyme. However, methotrexate or aminopterin, which are confirmed inhibitors of Arabidopsis dihydrofolate reductase, decreased the flux through the GDC/SHMT system by 60% after 48 h and by 100% in 96 h. The uptake of [α-13C]Gly was not inhibited by either drug class. The specificity of methotrexate action was shown by the ability of 5-formyl-THF to restore flux through the GDC/SHMT pathway in methotrexate-inhibited plants. The experiments with sulfonamides strongly suggest that the mitochondrial THF pool has a long half-life. The studies with methotrexate support the additional, critical role of dihydrofolate reductase in recycling THF oxidized in thymidylate synthesis.

Long-term, progressive hippocampal cell loss and dysfunction induced by early-life administration of corticotropin-releasing hormone reproduce the effects of early-life stress

Stress early in postnatal life may result in long-term memory deficits and selective loss of hippocampal neurons. The mechanisms involved are poorly understood, but they may involve molecules and processes in the immature limbic system that are activated by stressful challenges. We report that administration of corticotropin-releasing hormone (CRH), the key limbic stress modulator, to the brains of immature rats reproduced the consequences of early-life stress, reducing memory functions throughout life. These deficits were associated with progressive loss of hippocampal CA3 neurons and chronic up-regulation of hippocampal CRH expression. Importantly, they did not require the presence of stress levels of glucocorticoids. These findings indicate a critical role for CRH in the mechanisms underlying the long-term effects of early-life stress on hippocampal integrity and function.

Archaeal-eubacterial mergers in the origin of Eukarya: phylogenetic classification of life.

A symbiosis-based phylogeny leads to a consistent, useful classification system for all life. "Kingdoms" and "Domains" are replaced by biological names for the most inclusive taxa: Prokarya (bacteria) and Eukarya (symbiosis-derived nucleated organisms). The earliest Eukarya, anaerobic mastigotes, hypothetically originated from permanent whole-cell fusion between members of Archaea (e.g., Thermoplasma-like organisms) and of Eubacteria (e.g., Spirochaeta-like organisms). Molecular biology, life-history, and fossil record evidence support the reunification of bacteria as Prokarya while subdividing Eukarya into uniquely defined subtaxa: Protoctista, Animalia, Fungi, and Plantae.