Multi-dimensional classification of GABAergic interneurons with Bayesian network-modeled label uncertainty

Interneuron classification is an important and long-debated topic in neuroscience. A recent study provided a data set of digitally reconstructed interneurons classified by 42 leading neuroscientists according to a pragmatic classification scheme composed of five categorical variables, namely, of the interneuron type and four features of axonal morphology. From this data set we now learned a model which can classify interneurons, on the basis of their axonal morphometric parameters, into these five descriptive variables simultaneously. Because of differences in opinion among the neuroscientists, especially regarding neuronal type, for many interneurons we lacked a unique, agreed-upon classification, which we could use to guide model learning. Instead, we guided model learning with a probability distribution over the neuronal type and the axonal features, obtained, for each interneuron, from the neuroscientists’ classification choices. We conveniently encoded such probability distributions with Bayesian networks, calling them label Bayesian networks (LBNs), and developed a method to predict them. This method predicts an LBN by forming a probabilistic consensus among the LBNs of the interneurons most similar to the one being classified. We used 18 axonal morphometric parameters as predictor variables, 13 of which we introduce in this paper as quantitative counterparts to the categorical axonal features. We were able to accurately predict interneuronal LBNs. Furthermore, when extracting crisp (i.e., non-probabilistic) predictions from the predicted LBNs, our method outperformed related work on interneuron classification. Our results indicate that our method is adequate for multi-dimensional classification of interneurons with probabilistic labels. Moreover, the introduced morphometric parameters are good predictors of interneuron type and the four features of axonal morphology and thus may serve as objective counterparts to the subjective, categorical axonal features.

Planning and management of mobility in natural protected areas

The final purpose is the identification of: a) the effects of various choices in transport planning, both at long term and strategic level; b) the most effective policies of mobility management. The preliminary work was articulated in the following steps: 1) definition of protected area on the basis of ecological and socio-economic criteria and legislative constraints; 2) analysis of mobility needs in the protected areas; 3) reconstruction of the state of the art of mobility management in natural parks at European level; 4) analysis of used traffic flows measurement methods; 5) analysis of environmental impacts due to transport systems modelling (limited to air pollution and noise); 6) identification of mitigation measures to the potentially applied. The whole methodology has been firstly tested on the case study of the National Park of ?Gran Sasso and Monti della Laga? and further validated on the National Park of ?Gargano?, both located Italy: i) the concerned area has been zoned according to the land-use peculiarities; ii) the local situations of transport infrastructure (roads and parking), services (public transport systems) and rules (traffic regulations) have been mapped with references to physical and functional attributes; iii) the mobility, both systematic and touristic, has been synthetically represented in an origin-destination matrix. By means of an assignment model it has been determined the distribution of flows and the corresponding average speeds to quantify gaseous and noise emissions. On this basis the environmental criticalities in the reference scenario have been highlighted, as well as some alternative scenarios including both operational and infrastructural measures have been identified. The comparison between the projects and the reference scenario allowed the quantification of the effects (variation of emissions) for each scenario and a selection of the most effective management actions to be taken.

Propuesta de un marco conceptual para el análisis comparativo de las redes de distribución de dos supermercados online = Proposal of a conceptual framework for the comparative analysis of the distribution networks of two online supermarkets

In this article the network configuration for fulfillment and distribution of online orders of two British retailers is analyzed and compared. For this purpose, it is proposed a conceptual framework that consists of the key following aspects: network configuration, transportation management and location of demand. As a result is not obvious to determine the ideal centralization degree in each case. Finally, it is suggested the future development of an analytic tool that helps to choose the most appropriate model.

Modelo para la integración del trazado de infraestructuras lineales en el paisaje y el medio ambiente basado en SIG

El análisis de las diferentes alternativas en la planificación y diseño de corredores y trazados de carreteras debe basarse en la correcta definición de variables territoriales que sirvan como criterios para la toma de decisión y esto requiere un análisis ambiental preliminar de esas variables de calidad. En España, los estudios de viabilidad de nuevas carreteras y autovías están asociados a una fase del proceso de decisión que se corresponde con el denominado Estudio Informativo, el cual establece condicionantes físicos, ambientales, de uso del suelo y culturales que deben ser considerados en las primeras fases de la definición del trazado de un corredor de carretera. Así, la metodología más frecuente es establecer diferentes niveles de capacidad de acogida del territorio en el área de estudio con el fin de resumir las variables territoriales en mapas temáticos y facilitar el proceso de trazado de las alternativas de corredores de carretera. El paisaje es un factor limitante a tener en cuenta en la planificación y diseño de carreteras y, por tanto, deben buscarse trazados más sostenibles en relación con criterios estéticos y ecológicos del mismo. Pero este factor no es frecuentemente analizado en los Estudios Informativos e incluso, si es considerado, los estudios específicos de la calidad del paisaje (estético y ecológico) y de las formas del terreno no incorporan las recomendaciones de las guías de trazado para evitar o reducir los impactos en el paisaje. Además, los mapas de paisaje que se generan en este tipo de estudios no se corresponden con la escala de desarrollo del Estudio Informativo (1:5.000). Otro déficit común en planificación de corredores y trazados de carreteras es que no se tiene en cuenta la conectividad del paisaje durante el proceso de diseño de la carretera para prevenir la afección a los corredores de fauna existentes en el paisaje. Este déficit puede originar un posterior efecto barrera en los movimientos dispersivos de la fauna y la fragmentación de sus hábitats debido a la ocupación parcial o total de las teselas de hábitats con importancia biológica para la fauna (o hábitats focales) y a la interrupción de los corredores de fauna que concentran esos movimientos dispersivos de la fauna entre teselas. El objetivo principal de esta tesis es mejorar el estudio del paisaje para prevenir su afección durante el proceso de trazado de carreteras, facilitar la conservación de los corredores de fauna (o pasillos verdes) y la localización de medidas preventivas y correctoras en términos de selección y cuantificación de factores de idoneidad a fin de reducir los impactos visuales y ecológicos en el paisaje a escala local. Concretamente, la incorporación de valores cuantitativos y bien justificados en el proceso de decisión permite incrementar la transparencia en el proceso de diseño de corredores y trazados de carreteras. Con este fin, se han planteado cuatro preguntas específicas en esta investigación (1) ¿Cómo se seleccionan y evalúan los factores territoriales limitantes para localizar una nueva carretera por los profesionales españoles de planificación del territorio en relación con el paisaje? (2) ¿Cómo pueden ser definidos los corredores de fauna a partir de factores del paisaje que influyen en los movimientos dispersivos de la fauna? (3) ¿Cómo pueden delimitarse y evaluarse los corredores de fauna incluyendo el comportamiento parcialmente errático en los movimientos dispersivos de la fauna y el efecto barrera de los elementos antrópicos a una escala local? (4) ¿Qué y cómo las recomendaciones de diseño de carreteras relacionadas con el paisaje y las formas del terreno pueden ser incluidas en un modelo de Sistemas de Información Geográfica (SIG) para ayudar a los ingenieros civiles durante el proceso de diseño de un trazado de carreteras bajo el punto de vista de la sostenibilidad?. Esta tesis doctoral propone nuevas metodologías que mejoran el análisis visual y ecológico del paisaje utilizando indicadores y modelos SIG para obtener alternativas de trazado que produzcan un menor impacto en el paisaje. Estas metodologías fueron probadas en un paisaje heterogéneo con una alta tasa de densidad de corzo (Capreolus capreolus L.), uno de los grandes mamíferos más atropellados en la red de carreteras españolas, y donde está planificada la construcción de una nueva autovía que atravesará la mitad del área de distribución del corzo. Inicialmente, se han analizado las variables utilizadas en 22 estudios de proyectos de planificación de corredores de carreteras promovidos por el Ministerio de Fomento entre 2006 y 2008. Estas variables se agruparon según condicionantes físicos, ambientales, de usos del suelo y culturales con el fin de comparar los valores asignados de capacidad de acogida del territorio a cada variable en los diferentes estudios revisados. Posteriormente, y como etapa previa de un análisis de conectividad, se construyó un mapa de resistencia de los movimientos dispersivos del corzo en base a la literatura y al juicio de expertos. Usando esta investigación como base, se le asignó un valor de resistencia a cada factor seleccionado para construir la matriz de resistencia, ponderándolo y combinándolo con el resto de factores usando el proceso analítico jerárquico y los operadores de lógica difusa como métodos de análisis multicriterio. Posteriormente, se diseñó una metodología SIG para delimitar claramente la extensión física de los corredores de fauna de acuerdo a un valor umbral de ancho geométrico mínimo, así como la existencia de múltiples potenciales conexiones entre cada par de teselas de hábitats presentes en el paisaje estudiado. Finalmente, se realizó un procesado de datos Light Detection and Ranging (LiDAR) y un modelo SIG para calcular la calidad del paisaje (estético y ecológico), las formas del terreno que presentan características similares para trazar una carretera y la acumulación de vistas de potenciales conductores y observadores de los alrededores de la nueva vía. Las principales contribuciones de esta investigación al conocimiento científico existente en el campo de la evaluación del impacto ambiental en relación al diseño de corredores y trazados de carreteras son cuatro. Primero, el análisis realizado de 22 Estudios Informativos de planificación de carreteras reveló que los métodos aplicados por los profesionales para la evaluación de la capacidad de acogida del territorio no fue suficientemente estandarizada, ya que había una falta de uniformidad en el uso de fuentes cartográficas y en las metodologías de evaluación de la capacidad de acogida del territorio, especialmente en el análisis de la calidad del paisaje estético y ecológico. Segundo, el análisis realizado en esta tesis destaca la importancia de los métodos multicriterio para estructurar, combinar y validar factores que limitan los movimientos dispersivos de la fauna en el análisis de conectividad. Tercero, los modelos SIG desarrollados Generador de alternativas de corredores o Generator of Alternative Corridors (GAC) y Eliminador de Corredores Estrechos o Narrow Corridor Eraser (NCE) pueden ser aplicados sistemáticamente y sobre una base científica en análisis de conectividad como una mejora de las herramientas existentes para la comprensión el paisaje como una red compuesta por nodos y enlaces interconectados. Así, ejecutando los modelos GAC y NCE de forma iterativa, pueden obtenerse corredores alternativos con similar probabilidad de ser utilizados por la fauna y sin que éstos presenten cuellos de botella. Cuarto, el caso de estudio llevado a cabo de prediseño de corredores y trazado de una nueva autovía ha sido novedoso incluyendo una clasificación semisupervisada de las formas del terreno, filtrando una nube de puntos LiDAR e incluyendo la nueva geometría 3D de la carretera en el Modelo Digital de Superficie (MDS). El uso combinado del procesamiento de datos LiDAR y de índices y clasificaciones geomorfológicas puede ayudar a los responsables encargados en la toma de decisiones a evaluar qué alternativas de trazado causan el menor impacto en el paisaje, proporciona una visión global de los juicios de valor más aplicados y, en conclusión, define qué medidas de integración paisajística correctoras deben aplicarse y dónde. ABSTRACT The assessment of different alternatives in road-corridor planning and layout design must be based on a number of well-defined territorial variables that serve as decision-making criteria, and this requires a high-quality preliminary environmental analysis of those quality variables. In Spain, feasibility studies for new roads and motorways are associated to a phase of the decision procedure which corresponds with the one known as the Informative Study, which establishes the physical, environmental, land-use and cultural constraints to be considered in the early stages of defining road corridor layouts. The most common methodology is to establish different levels of Territorial Carrying Capacity (TCC) in the study area in order to summarize the territorial variables on thematic maps and facilitate the tracing process of road-corridor layout alternatives. Landscape is a constraint factor that must be considered in road planning and design, and the most sustainable layouts should be sought based on aesthetic and ecological criteria. However this factor is not often analyzed in Informative Studies and even if it is, baseline studies on landscape quality (aesthetic and ecological) and landforms do not usually include the recommendations of road tracing guides designed to avoid or reduce impacts on the landscape. The resolution of the landscape maps produced in this type of studies does not comply with the recommended road design scale (1:5,000) in the regulations for the Informative Study procedure. Another common shortcoming in road planning is that landscape ecological connectivity is not considered during road design in order to avoid affecting wildlife corridors in the landscape. In the prior road planning stage, this issue could lead to a major barrier effect for fauna dispersal movements and to the fragmentation of their habitat due to the partial or total occupation of habitat patches of biological importance for the fauna (or focal habitats), and the interruption of wildlife corridors that concentrate fauna dispersal movements between patches. The main goal of this dissertation is to improve the study of the landscape and prevent negative effects during the road tracing process, and facilitate the preservation of wildlife corridors (or green ways) and the location of preventive and corrective measures by selecting and quantifying suitability factors to reduce visual and ecological landscape impacts at a local scale. Specifically the incorporation of quantitative and well-supported values in the decision-making process provides increased transparency in the road corridors and layouts design process. Four specific questions were raised in this research: (1) How are territorial constraints selected and evaluated in terms of landscape by Spanish land-planning practitioners before locating a new road? (2) How can wildlife corridors be defined based on the landscape factors influencing the dispersal movements of fauna? (3) How can wildlife corridors be delimited and assessed to include the partially erratic movements of fauna and the barrier effect of the anthropic elements at a local scale? (4) How recommendations of road design related to landscape and landforms can be included in a Geographic Information System (GIS) model to aid civil engineers during the road layout design process and support sustainable development? This doctoral thesis proposes new methodologies that improve the assessment of the visual and ecological landscape character using indicators and GIS models to obtain road layout alternatives with a lower impact on the landscape. These methodologies were tested on a case study of a heterogeneous landscape with a high density of roe deer (Capreolus capreolus L.) –one of the large mammals most commonly hit by vehicles on the Spanish road network– and where a new motorway is planned to pass through the middle of their distribution area. We explored the variables used in 22 road-corridor planning projects sponsored by the Ministry of Public Works between 2006 and 2008. These variables were grouped into physical, environmental, land-use and cultural constraints for the purpose of comparing the TCC values assigned to each variable in the various studies reviewed. As a prior stage in a connectivity analysis, a map of resistance to roe deer dispersal movements was created based on the literature and experts judgment. Using this research as a base, each factor selected to build the matrix was assigned a resistance value and weighted and combined with the rest of the factors using the analytic hierarchy process (AHP) and fuzzy logic operators as multicriteria assessment (MCA) methods. A GIS methodology was designed to clearly delimit the physical area of wildlife corridors according to a geometric threshold width value, and the multiple potential connections between each pair of habitat patches in the landscape. A Digital Surface Model Light Detection and Ranging (LiDAR) dataset processing and a GIS model was performed to determine landscape quality (aesthetic and ecological) and landforms with similar characteristics for the road layout, and the cumulative viewshed of potential drivers and observers in the area surrounding the new motorway. The main contributions of this research to current scientific knowledge in the field of environmental impact assessment for road corridors and layouts design are four. First, the analysis of 22 Informative Studies on road planning revealed that the methods applied by practitioners for assessing the TCC were not sufficiently standardized due to the lack of uniformity in the cartographic information sources and the TCC valuation methodologies, especially in the analysis of the aesthetic and ecological quality of the landscape. Second, the analysis in this dissertation highlights the importance of multicriteria methods to structure, combine and validate factors that constrain wildlife dispersal movements in the connectivity analysis. Third, the “Generator of Alternative Corridors (GAC)” and “Narrow Corridor Eraser (NCE)” GIS models developed can be applied systematically and on a scientific basis in connectivity analyses to improve existing tools and understand landscape as a network composed of interconnected nodes and links. Thus, alternative corridors with similar probability of use by fauna and without bottlenecks can be obtained by iteratively running GAC and NCE models. Fourth, our case study of new motorway corridors and layouts design innovatively included semi-supervised classification of landforms, filtering of LiDAR point clouds and new 3D road geometry on the Digital Surface Model (DSM). The combined used of LiDAR data processing and geomorphological indices and classifications can help decision-makers assess which road layouts produce lower impacts on the landscape, provide an overall insight into the most commonly applied value judgments, and in conclusion, define which corrective measures should be applied in terms of landscaping, and where.

Evaluación de la distribución de riesgos en los distintos modelos de asociación público-privada para la provisión de infraestructura carretera en México

México es de los pocos países en el mundo que ha realizado dos grandes programas para la construcción de autopistas en colaboración con el sector privado. El primero, fue realizado entre 1989 y 1994, con resultados adversos por el mal diseño del esquema de concesiones; y, el segundo con mejores resultados, en operación desde 2003 mediante nuevos modelos de asociación público-privada (APP). El objetivo de la presente investigación es estudiar los modelos de asociación público-privada empleados en México para la provisión de infraestructura carretera, realizando el análisis y la evaluación de la distribución de riesgos entre el sector público y privado en cada uno de los modelos con el propósito de establecer una propuesta de reasignación de riesgos para disminuir el costo global y la incertidumbre de los proyectos. En la primera parte se describe el estado actual del conocimiento de las asociaciones público-privadas para desarrollar proyectos de infraestructura, incluyendo los antecedentes, la definición y las tipologías de los esquemas APP, así como la práctica internacional de programas como el modelo británico Private Finance Initiative (PFI), resultados de proyectos en la Unión Europea y programas APP en otros países. También, se destaca la participación del sector privado en el financiamiento de la infraestructura del transporte de México en la década de 1990. En los capítulos centrales se aborda el estudio de los modelos APP que se han utilizado en el país en la construcción de la red de carreteras de alta capacidad. Se presentan las características y los resultados del programa de autopistas 1989-94, así como el rescate financiero y las medidas de reestructuración de los proyectos concesionados, aspectos que obligaron a las autoridades mexicanas a cambiar la normatividad para la aprobación de los proyectos según su rentabilidad, modificar la legislación de caminos y diseñar nuevos esquemas de colaboración entre el gobierno y el sector privado. Los nuevos modelos APP vigentes desde 2003 son: nuevo modelo de concesiones para desarrollar autopistas de peaje, modelo de proyectos de prestación de servicios (peaje sombra) para modernizar carreteras existentes y modelo de aprovechamiento de activos para concesionar autopistas de peaje en operación a cambio de un pago. De estos modelos se realizaron estudios de caso en los que se determinan medidas de desempeño operativo (niveles de tráfico, costos y plazos de construcción) y rentabilidad financiera (tasa interna de retorno y valor presente neto). En la última parte se efectúa la identificación, análisis y evaluación de los riesgos que afectaron los costos, el tiempo de ejecución y la rentabilidad de los proyectos de ambos programas. Entre los factores de riesgo analizados se encontró que los más importantes fueron: las condiciones macroeconómicas del país (inflación, producto interno bruto, tipo de cambio y tasa de interés), deficiencias en la planificación de los proyectos (diseño, derecho de vía, tarifas, permisos y estimación del tránsito) y aportaciones públicas en forma de obra. Mexico is one of the few countries in the world that has developed two major programs for highway construction in collaboration with the private sector. The first one was carried out between 1989 and 1994 with adverse outcomes due to the wrong design of concession schemes; and, the second one, in operation since 2003, through new public-private partnership models (PPPs). The objective of this research is to study public-private partnership models used in Mexico for road infrastructure provision, performing the analysis and evaluation of risk’s distribution between the public and the private sector in each model in order to draw up a proposal for risk’s allocation to reduce the total cost and the uncertainty of projects. The first part describes the current state of knowledge in public-private partnership to develop infrastructure projects, including the history, definition and types of PPP models, as well as international practice of programs such as the British Private Finance Initiative (PFI) model, results in the European Union and PPP programs in other countries. Also, it stands out the private sector participation in financing of Mexico’s transport infrastructure in 1990s. The next chapters present the study of public-private partnerships models that have been used in the country in the construction of the high capacity road network. Characteristics and outcomes of the highway program 1989-94 are presented, as well as the financial bailout and restructuring measures of the concession projects, aspects that forced the Mexican authorities to change projects regulations, improve road’s legislation and design new schemes of cooperation between the Government and the private sector. The new PPP models since 2003 are: concession model to develop toll highways, private service contracts model (shadow toll) to modernize existing roads and highway assets model for the concession of toll roads in operation in exchange for a payment. These models were analyzed using case studies in which measures of operational performance (levels of traffic, costs and construction schedules) and financial profitability (internal rate of return and net present value) are determined. In the last part, the analysis and assessment of risks that affect costs, execution time and profitability of the projects are carried out, for both programs. Among the risk factors analyzed, the following ones were found to be the most important: country macroeconomic conditions (inflation, gross domestic product, exchange rate and interest rate), deficiencies in projects planning (design, right of way, tolls, permits and traffic estimation) and public contributions in the form of construction works.

Red continua de espacios públicos verdes a escala municipal: el caso de Madrid

La ubicación marginal y el aislamiento de los Parques Urbanos implican infrautilización y deterioro de los mismos. Su interconexión mediante una red verde peatonal con limitaciones al tráfico rodado, supone una puesta en valor de todo el sistema de espacios libres y zonas verdes de la ciudad y un importante incentivo para desviar determinados flujos rodados a otras vías cercanas, rescatando así, espacios urbanos a favor del peatón. Esta conexión ayuda a minimizar la fragmentación del espacio urbano, generando nexos de unión, que articulan y estructuran la ciudad. En esta investigación se describe una metodología para localizar y seleccionar conectores entre distintos Parques Urbanos, evaluando su funcionalidad, seguridad, confort y atractivo. Como caso de estudio, se proponen las conexiones entre los Parques Públicos de una entidad determinada, situados en la ciudad de Madrid. Esta metodología es elástica, universal, y dada su sencillez, aplicable por personas no especializadas; estas características la hacen especialmente valiosa, para apoyar la toma de decisiones en los instrumentos de Ordenación del Territorio y en el desarrollo de Planes Específicos encaminados a la mejora de la calidad ambiental y al aumento de la potencialidad peatonal de la ciudad. ABSTRACT The marginal location and the solitary distribution of City Parks are factors that expose these “City Lungs” to be underused and damaged. The City Parks chained by a green pedestrian web with a minimized relevance for engine vehicles, generates an important enhancement for open spaces and green areas of the City, and a crucial incentive for altering the course of vehicle transportation from particular areas to proximate avenues and arteries, resulting in urban spaces being exclusively for pedestrians. This network helps to minimize the fragmentation of the urban space generating nodes of connection that provide a structure to the city. In this investigation a procedure is described for the purpose of identifying and prioritizing green connectors amongst the City Parks, evaluating the functionality, security, comfort, and attractions. For this research paper, it is proposed to establish a green web, which links up the main City Parks of Madrid. This methodology is elastic, universal, and due to its simplicity, can be applicable for non-specialized people of the field: these features make this urban concept a very estimable and valued idea that can be taken into consideration for future urban planning and for the development of specific plans with the object of improving the quality of the environment and lead to an increase of the pedestrian potential within the city.

Mapping species distributions : A comparison of skilled naturalist and lay citizen science recording

Acknowledgements We are grateful to Elaine O’Mahony, Imogen Pearce, Richard Comont, Anthony McCluskey and other BBCT staff for the many hours of BeeWatch species identification and for all people who submitted sightings to BeeWatch, OPAL, BWARS and the various local recording schemes and societies. We thank the NBN for allowing us to download the bumblebee records without strings attached, and the Essex, Greater London, Cumbria and Sussex based recording centres for providing records upon request. Finally, we are indebted to Tom August and two anonymous reviewers for their valuable critique on an earlier version of this work.

High-Affinity Binding Of The AP-1 Adaptor Complex to Trans-Golgi Network Membranes Devoid Of Mannose 6-Phosphate Receptors

The GTP-binding protein ADP-ribosylation factor (ARF) initiates clathrin-coat assembly at the trans-Goli network (TGN) by generating high-affinity membrane-binding sites for the AP-1 adaptor complex. Both transmembrane proteins, which are sorted into the assembling coated bud, and novel docking proteins have been suggested to be partners with GTP-bound ARF in generating the AP-1-docking sites. The best characterized, and probably the major transmembrane molecules sorted into the clathrin-coated vesicles that form on the TGN, are the mannose 6-phosphate receptors (MPRs). Here, we have examined the role of the MPRs in the AP-1 recruitment process by comparing fibroblasts derived from embryos of either normal or MPR-negative animals. Despite major alterations to the lysosome compartment in the MPR-deficient cells, the steady-state distribution of AP-1 at the TGN is comparable to that of normal cells. Golgi-enriched membranes prepared from the receptor-negative cells also display an apparently normal capacity to recruit AP-1 in vitro in the presence of ARF and either GTP or GTPγS. The AP-1 adaptor is recruited specifically onto the TGN and not onto the numerous abnormal membrane elements that accumulate within the MPR-negative fibroblasts. AP-1 bound to TGN membranes from either normal or MPR-negative fibroblasts is fully resistant to chemical extraction with 1 M Tris-HCl, pH 7, indicating that the adaptor binds to both membrane types with high affinity. The only difference we do note between the Golgi prepared from the MPR-deficient cells and the normal cells is that AP-1 recruited onto the receptor-lacking membranes in the presence of ARF1·GTP is consistently more resistant to extraction with Tris. Because sensitivity to Tris extraction correlates well with nucleotide hydrolysis, this finding might suggest a possible link between MPR sorting and ARF GAP regulation. We conclude that the MPRs are not essential determinants in the initial steps of AP-1 binding to the TGN but, instead, they may play a regulatory role in clathrin-coated vesicle formation by affecting ARF·GTP hydrolysis.

Apiconuclear Organization of Microtubules Does Not Specify Protein Delivery from the Trans-Golgi Network to Different Membrane Domains in Polarized Epithelial Cells

In nonpolarized epithelial cells, microtubules originate from a broad perinuclear region coincident with the distribution of the Golgi complex and extend outward to the cell periphery (perinuclear [PN] organization). During development of epithelial cell polarity, microtubules reorganize to form long cortical filaments parallel to the lateral membrane, a meshwork of randomly oriented short filaments beneath the apical membrane, and short filaments at the base of the cell; the Golgi becomes localized above the nucleus in the subapical membrane cytoplasm (apiconuclear [AN] organization). The AN-type organization of microtubules is thought to be specialized in polarized epithelial cells to facilitate vesicle trafficking between the trans-Golgi Network (TGN) and the plasma membrane. We describe two clones of MDCK cells, which have different microtubule distributions: clone II/G cells, which gradually reorganize a PN-type distribution of microtubules and the Golgi complex to an AN-type during development of polarity, and clone II/J cells which maintain a PN-type organization. Both cell clones, however, exhibit identical steady-state polarity of apical and basolateral proteins. During development of cell surface polarity, both clones rapidly establish direct targeting pathways for newly synthesized gp80 and gp135/170, and E-cadherin between the TGN and apical and basolateral membrane, respectively; this occurs before development of the AN-type microtubule/Golgi organization in clone II/G cells. Exposure of both clone II/G and II/J cells to low temperature and nocodazole disrupts >99% of microtubules, resulting in: 1) 25–50% decrease in delivery of newly synthesized gp135/170 and E-cadherin to the apical and basolateral membrane, respectively, in both clone II/G and II/J cells, but with little or no missorting to the opposite membrane domain during all stages of polarity development; 2) ∼40% decrease in delivery of newly synthesized gp80 to the apical membrane with significant missorting to the basolateral membrane in newly established cultures of clone II/G and II/J cells; and 3) variable and nonspecific delivery of newly synthesized gp80 to both membrane domains in fully polarized cultures. These results define several classes of proteins that differ in their dependence on intact microtubules for efficient and specific targeting between the Golgi and plasma membrane domains.

Vesicle-associated Membrane Protein 4 is Implicated in Trans-Golgi Network Vesicle Trafficking

The trans-Golgi network (TGN) plays a pivotal role in directing proteins in the secretory pathway to the appropriate cellular destination. VAMP4, a recently discovered member of the vesicle-associated membrane protein (VAMP) family of trafficking proteins, has been suggested to play a role in mediating TGN trafficking. To better understand the function of VAMP4, we examined its precise subcellular distribution. Indirect immunofluorescence and electron microscopy revealed that the majority of VAMP4 localized to tubular and vesicular membranes of the TGN, which were in part coated with clathrin. In these compartments, VAMP4 was found to colocalize with the putative TGN-trafficking protein syntaxin 6. Additional labeling was also present on clathrin-coated and noncoated vesicles, on endosomes and the medial and trans side of the Golgi complex, as well as on immature secretory granules in PC12 cells. Immunoprecipitation of VAMP4 from rat brain detergent extracts revealed that VAMP4 exists in a complex containing syntaxin 6. Converging lines of evidence implicate a role for VAMP4 in TGN-to-endosome transport.

Fault location in power distribution networks with distributed generation.

This research presents the development and implementation of fault location algorithms in power distribution networks with distributed generation units installed along their feeders. The proposed algorithms are capable of locating the fault based on voltage and current signals recorded by intelligent electronic devices installed at the end of the feeder sections, information to compute the loads connected to these feeders and their electric characteristics, and the operating status of the network. In addition, this work presents the study of analytical models of distributed generation and load technologies that could contribute to the performance of the proposed fault location algorithms. The validation of the algorithms was based on computer simulations using network models implemented in ATP, whereas the algorithms were implemented in MATLAB.

Breaking down complex saproxylic communities: understanding sub-networks structure and implications to network robustness

Saproxylic insect communities inhabiting tree hollow microhabitats correspond with large food webs which simultaneously are constituted by multiple types of plant-animal and animal-animal interactions, according to the use of trophic resources (wood- and insect-dependent sub-networks), or to trophic habits or interaction types (xylophagous, saprophagous, xylomycetophagous, predators and commensals). We quantitatively assessed which properties of specialised networks were present in a complex networks involving different interacting types such as saproxylic community, and how they can be organised in trophic food webs. The architecture, interacting patterns and food web composition were evaluated along sub-networks, analysing their implications to network robustness from random and directed extinction simulations. A structure of large and cohesive modules with weakly connected nodes was observed throughout saproxylic sub-networks, composing the main food webs constituting this community. Insect-dependent sub-networks were more modular than wood-dependent sub-networks. Wood-dependent sub-networks presented higher species degree, connectance, links, linkage density, interaction strength, and were less specialised and more aggregated than insect-dependent sub-networks. These attributes defined high network robustness in wood-dependent sub-networks. Finally, our results emphasise the relevance of modularity, differences among interacting types and interrelations among them in modelling the structure of saproxylic communities and in determining their stability.

A methodology for evacuation route planning inside buildings using geospatial technology

Evacuation route planning is a fundamental task for building engineering projects. Safety regulations are established so that all occupants are driven on time out of a building to a secure place when faced with an emergency situation. As an example, Spanish building code requires the planning of evacuation routes on large and, usually, public buildings. Engineers often plan these routes on single building projects, repeatedly assigning clusters of rooms to each emergency exit in a trial-and-error process. But problems may arise for a building complex where distribution and use changes make visual analysis cumbersome and sometimes unfeasible. This problem could be solved by using well-known spatial analysis techniques, implemented as a specialized software able to partially emulate engineer reasoning. In this paper we propose and test an easily reproducible methodology that makes use of free and open source software components for solving a case study. We ran a complete test on a building floor at the University of Alicante (Spain). This institution offers a web service (WFS) that allows retrieval of 2D geometries from any building within its campus. We demonstrate how geospatial technologies and computational geometry algorithms can be used for automating the creation and optimization of evacuation routes. In our case study, the engineers’ task is to verify that the load capacity of each emergency exit does not exceed the standards specified by Spain’s current regulations. Using Dijkstra’s algorithm, we obtain the shortest paths from every room to the most appropriate emergency exit. Once these paths are calculated, engineers can run simulations and validate, based on path statistics, different cluster configurations. Techniques and tools applied in this research would be helpful in the design and risk management phases of any complex building project.

Differences and connections: beyond universal theories in planning, urban, and heritage studies

The annual Young Academics network of the Association of European School of Planning (AESOP YA) conference, entitled Differences and Connections, was held for the first time in a Southern Italian city, Palermo, Italy, during 23–26 March 2015. The call for papers attracted a wide range of authors within the field of planning and other related fields. Forty-five contributions by young academic scholars, representing 19 countries, were selected by the conference scientific committee to critically explore the themes of the conference.

Analyse de la distribution d'Escherichia coli, utilisée comme un marqueur microbiologique, dans un réseau de production porcin

La présence d’Escherichia coli pathogènes en élevages porcins entraine des retards de croissance et la mortalité. La transmission des E. coli pathogènes entre les élevages et l'abattoir d’un même réseau de production n'est pas bien décrite. La détection des gènes de virulence des E. coli pathogènes pourrait permettre d’identifier un marqueur de contamination dans le réseau. L’objectif de cette étude a été d’identifier un marqueur de contamination E. coli dans un réseau de production porcine défini afin de décrire certains modes de transmission des E. coli pathogènes. Pour ce faire, une région géographique comprenant 10 fermes d’engraissement, un abattoir et un réseau de transport a été sélectionnée. Trois lots de production consécutifs par ferme ont été suivis pendant 12 mois. Des échantillons environnementaux ont été prélevés à l’intérieur et à l’extérieur des fermes (3 visites d’élevage), dans la cour de l’abattoir (2 visites lors de sorties de lot) et sur le camion de transport. La détection des gènes de virulence (eltB, estA, estB, faeG, stxA, stx2A, eae, cnf, papC, iucD, tsh, fedA) dans les échantillons a été réalisée par PCR multiplexe conventionnelle. La distribution temporelle et spatiale des gènes de virulence a permis d’identifier le marqueur de contamination ETEC/F4 défini par la détection d’au moins un gène d’entérotoxine ETEC (estB, estA et eltB) en combinaison avec le gène de l’adhésine fimbriaire (faeG). La distribution des échantillons positifs ETEC/F4 qualifie la cour de l’abattoir comme un réservoir de contamination fréquenté par les transporteurs, vecteurs de contamination entre les élevages. Ceci suggère le lien microbiologique entre l’élevage, les transporteurs et l’abattoir jouant chacun un rôle dans la dissémination des microorganismes pathogènes et potentiellement zoonotiques en production porcine.