Modelling the Impact of Built Environment, Geographical Scales and Latent Constructs On Individual Travel Behaviour

La relación entre la estructura urbana y la movilidad ha sido estudiada desde hace más de 70 años. El entorno urbano incluye múltiples dimensiones como por ejemplo: la estructura urbana, los usos de suelo, la distribución de instalaciones diversas (comercios, escuelas y zonas de restauración, parking, etc.). Al realizar una revisión de la literatura existente en este contexto, se encuentran distintos análisis, metodologías, escalas geográficas y dimensiones, tanto de la movilidad como de la estructura urbana. En este sentido, se trata de una relación muy estudiada pero muy compleja, sobre la que no existe hasta el momento un consenso sobre qué dimensión del entorno urbano influye sobre qué dimensión de la movilidad, y cuál es la manera apropiada de representar esta relación. Con el propósito de contestar estas preguntas investigación, la presente tesis tiene los siguientes objetivos generales: (1) Contribuir al mejor entendimiento de la compleja relación estructura urbana y movilidad. y (2) Entender el rol de los atributos latentes en la relación entorno urbano y movilidad. El objetivo específico de la tesis es analizar la influencia del entorno urbano sobre dos dimensiones de la movilidad: número de viajes y tipo de tour. Vista la complejidad de la relación entorno urbano y movilidad, se pretende contribuir al mejor entendimiento de la relación a través de la utilización de 3 escalas geográficas de las variables y del análisis de la influencia de efectos inobservados en la movilidad. Para el análisis se utiliza una base de datos conformada por tres tipos de datos: (1) Una encuesta de movilidad realizada durante los años 2006 y 2007. Se obtuvo un total de 943 encuestas, en 3 barrios de Madrid: Chamberí, Pozuelo y Algete. (2) Información municipal del Instituto Nacional de Estadística: dicha información se encuentra enlazada con los orígenes y destinos de los viajes recogidos en la encuesta. Y (3) Información georeferenciada en Arc-GIS de los hogares participantes en la encuesta: la base de datos contiene información respecto a la estructura de las calles, localización de escuelas, parking, centros médicos y lugares de restauración. Se analizó la correlación entre e intra-grupos y se modelizaron 4 casos de atributos bajo la estructura ordinal logit. Posteriormente se evalúa la auto-selección a través de la estimación conjunta de las elecciones de tipo de barrio y número de viajes. La elección del tipo de barrio consta de 3 alternativas: CBD, Urban y Suburban, según la zona de residencia recogida en las encuestas. Mientras que la elección del número de viajes consta de 4 categorías ordinales: 0 viajes, 1-2 viajes, 3-4 viajes y 5 o más viajes. A partir de la mejor especificación del modelo ordinal logit. Se desarrolló un modelo joint mixed-ordinal conjunto. Los resultados indican que las variables exógenas requieren un análisis exhaustivo de correlaciones con el fin de evitar resultados sesgados. ha determinado que es importante medir los atributos del BE donde se realiza el viaje, pero también la información municipal es muy explicativa de la movilidad individual. Por tanto, la percepción de las zonas de destino a nivel municipal es considerada importante. En el contexto de la Auto-selección (self-selection) es importante modelizar conjuntamente las decisiones. La Auto-selección existe, puesto que los parámetros estimados conjuntamente son significativos. Sin embargo, sólo ciertos atributos del entorno urbano son igualmente importantes sobre la elección de la zona de residencia y frecuencia de viajes. Para analizar la Propensión al Viaje, se desarrolló un modelo híbrido, formado por: una variable latente, un indicador y un modelo de elección discreta. La variable latente se denomina “Propensión al Viaje”, cuyo indicador en ecuación de medida es el número de viajes; la elección discreta es el tipo de tour. El modelo de elección consiste en 5 alternativas, según la jerarquía de actividades establecida en la tesis: HOME, no realiza viajes durante el día de estudio, HWH tour cuya actividad principal es el trabajo o estudios, y no se realizan paradas intermedias; HWHs tour si el individuo reaiza paradas intermedias; HOH tour cuya actividad principal es distinta a trabajo y estudios, y no se realizan paradas intermedias; HOHs donde se realizan paradas intermedias. Para llegar a la mejor especificación del modelo, se realizó un trabajo importante considerando diferentes estructuras de modelos y tres tipos de estimaciones. De tal manera, se obtuvieron parámetros consistentes y eficientes. Los resultados muestran que la modelización de los tours, representa una ventaja sobre la modelización de los viajes, puesto que supera las limitaciones de espacio y tiempo, enlazando los viajes realizados por la misma persona en el día de estudio. La propensión al viaje (PT) existe y es específica para cada tipo de tour. Los parámetros estimados en el modelo híbrido resultaron significativos y distintos para cada alternativa de tipo de tour. Por último, en la tesis se verifica que los modelos híbridos representan una mejora sobre los modelos tradicionales de elección discreta, dando como resultado parámetros consistentes y más robustos. En cuanto a políticas de transporte, se ha demostrado que los atributos del entorno urbano son más importantes que los LOS (Level of Service) en la generación de tours multi-etapas. la presente tesis representa el primer análisis empírico de la relación entre los tipos de tours y la propensión al viaje. El concepto Propensity to Travel ha sido desarrollado exclusivamente para la tesis. Igualmente, el desarrollo de un modelo conjunto RC-Number of trips basado en tres escalas de medida representa innovación en cuanto a la comparación de las escalas geográficas, que no había sido hecha en la modelización de la self-selection. The relationship between built environment (BE) and travel behaviour (TB) has been studied in a number of cases, using several methods - aggregate and disaggregate approaches - and different focuses – trip frequency, automobile use, and vehicle miles travelled and so on. Definitely, travel is generated by the need to undertake activities and obtain services, and there is a general consensus that urban components affect TB. However researches are still needed to better understand which components of the travel behaviour are affected most and by which of the urban components. In order to fill the gap in the research, the present dissertation faced two main objectives: (1) To contribute to the better understanding of the relationship between travel demand and urban environment. And (2) To develop an econometric model for estimating travel demand with urban environment attributes. With this purpose, the present thesis faced an exhaustive research and computation of land-use variables in order to find the best representation of BE for modelling trip frequency. In particular two empirical analyses are carried out: 1. Estimation of three dimensions of travel demand using dimensions of urban environment. We compare different travel dimensions and geographical scales, and we measure self-selection contribution following the joint models. 2. Develop a hybrid model, integrated latent variable and discrete choice model. The implementation of hybrid models is new in the analysis of land-use and travel behaviour. BE and TB explicitly interact and allow richness information about a specific individual decision process For all empirical analysis is used a data-base from a survey conducted in 2006 and 2007 in Madrid. Spatial attributes describing neighbourhood environment are derived from different data sources: National Institute of Statistics-INE (Administrative: municipality and district) and GIS (circular units). INE provides raw data for such spatial units as: municipality and district. The construction of census units is trivial as the census bureau provides tables that readily define districts and municipalities. The construction of circular units requires us to determine the radius and associate the spatial information to our households. The first empirical part analyzes trip frequency by applying an ordered logit model. In this part is studied the effect of socio-economic, transport and land use characteristics on two travel dimensions: trip frequency and type of tour. In particular the land use is defined in terms of type of neighbourhoods and types of dwellers. Three neighbourhood representations are explored, and described three for constructing neighbourhood attributes. In particular administrative units are examined to represent neighbourhood and circular – unit representation. Ordered logit models are applied, while ordinal logit models are well-known, an intensive work for constructing a spatial attributes was carried out. On the other hand, the second empirical analysis consists of the development of an innovative econometric model that considers a latent variable called “propensity to travel”, and choice model is the choice of type of tour. The first two specifications of ordinal models help to estimate this latent variable. The latent variable is unobserved but the manifestation is called “indicators”, then the probability of choosing an alternative of tour is conditional to the probability of latent variable and type of tour. Since latent variable is unknown we fit the integral over its distribution. Four “sets of best variables” are specified, following the specification obtained from the correlation analysis. The results evidence that the relative importance of SE variables versus BE variables depends on how BE variables are measured. We found that each of these three spatial scales has its intangible qualities and drawbacks. Spatial scales play an important role on predicting travel demand due to the variability in measures at trip origin/destinations within the same administrative unit (municipality, district and so on). Larger units will produce less variation in data; but it does not affect certain variables, such as public transport supply, that are more significant at municipality level. By contrast, land-use measures are more efficient at district level. Self-selection in this context, is weak. Thus, the influence of BE attributes is true. The results of the hybrid model show that unobserved factors affect the choice of tour complexity. The latent variable used in this model is propensity to travel that is explained by socioeconomic aspects and neighbourhood attributes. The results show that neighbourhood attributes have indeed a significant impact on the choice of the type of tours either directly and through the propensity to travel. The propensity to travel has a different impact depending on the structure of each tour and increases the probability of choosing more complex tours, such as tours with many intermediate stops. The integration of choice and latent variable model shows that omitting important perception and attitudes leads to inconsistent estimates. The results also indicate that goodness of fit improves by adding the latent variable in both sequential and simultaneous estimation. There are significant differences in the sensitivity to the latent variable across alternatives. In general, as expected, the hybrid models show a major improvement into the goodness of fit of the model, compared to a classical discrete choice model that does not incorporate latent effects. The integrated model leads to a more detailed analysis of the behavioural process. Summarizing, the effect that built environment characteristics on trip frequency studied is deeply analyzed. In particular we tried to better understand how land use characteristics can be defined and measured and which of these measures do have really an impact on trip frequency. We also tried to test the superiority of HCM on this field. We can concluded that HCM shows a major improvement into the goodness of fit of the model, compared to classical discrete choice model that does not incorporate latent effects. And consequently, the application of HCM shows the importance of LV on the decision of tour complexity. People are more elastic to built environment attributes than level of services. Thus, policy implications must take place to develop more mixed areas, work-places in combination with commercial retails.

Methodology for the design of offshore wind farms

Although still in an early stage, offshore wind development is now characterized by a boom process. This leads to the necessity of applying an integral management model for the design of offshore wind facilities, being the purpose of the model to achieve technical, economical and environmental viability, all within a sustainable development framework. The foregoing led to the research project exposed in this paper, consisting of drawing up an offshore wind farms methodological proposal; this methodology has a global and/or general nature or point of view whilst searching for optimization of the overall process of operations leading to the design of this type of installations and establishing collated theoretical bases for the further development of management tools. This methodological proposal follows a classical engineering thought scheme: it begins with the alternatives study, and ends with the detailed design. With this in mind, the paper includes the following sections: introduction, methodology used for the research project, conditioning factors, methodological proposal for the design of offshore wind farms, checking the methodological proposal, and conclusions

Guidance for the development of accessibility evaluation tools following the Unified Software Development Process

Automated and semi-automated accessibility evaluation tools are key to streamline the process of accessibility assessment, and ultimately ensure that software products, contents, and services meet accessibility requirements. Different evaluation tools may better fit different needs and concerns, accounting for a variety of corporate and external policies, content types, invocation methods, deployment contexts, exploitation models, intended audiences and goals; and the specific overall process where they are introduced. This has led to the proliferation of many evaluation tools tailored to specific contexts. However, tool creators, who may be not familiar with the realm of accessibility and may be part of a larger project, lack any systematic guidance when facing the implementation of accessibility evaluation functionalities. Herein we present a systematic approach to the development of accessibility evaluation tools, leveraging the different artifacts and activities of a standardized development process model (the Unified Software Development Process), and providing templates of these artifacts tailored to accessibility evaluation tools. The work presented specially considers the work in progress in this area by the W3C/WAI Evaluation and Report Working Group (ERT WG)

The effect of boundary conditions in the numerical solution of 3-D thermoelastic problems

After the extensive research on the capabilities of the Boundary Integral Equation Method produced during the past years the versatility of its applications has been well founded. Maybe the years to come will see the in-depth analysis of several conflictive points, for example, adaptive integration, solution of the system of equations, etc. This line is clear in academic research. In this paper we comment on the incidence of the manner of imposing the boundary conditions in 3-D coupled problems. Here the effects are particularly magnified: in the first place by the simple model used (constant elements) and secondly by the process of solution, i.e. first a potential problem is solved and then the results are used as data for an elasticity problem. The errors add to both processes and small disturbances, unimportant in separated problems, can produce serious errors in the final results. The specific problem we have chosen is especially interesting. Although more general cases (i.e. transient)can be treated, here the domain integrals can be converted into boundary ones and the influence of the manner in which boundary conditions are applied will reflect the whole importance of the problem.

Using floating car data to analyse the effects of ITS measures and eco-driving

The road transportation sector is responsible for around 25% of total man-made CO2 emissions worldwide. Considerable efforts are therefore underway to reduce these emissions using several approaches, including improved vehicle technologies, traffic management and changing driving behaviour. Detailed traffic and emissions models are used extensively to assess the potential effects of these measures. However, if the input and calibration data are not sufficiently detailed there is an inherent risk that the results may be inaccurate. This article presents the use of Floating Car Data to derive useful speed and acceleration values in the process of traffic model calibration as a means of ensuring more accurate results when simulating the effects of particular measures. The data acquired includes instantaneous GPS coordinates to track and select the itineraries, and speed and engine performance extracted directly from the on-board diagnostics system. Once the data is processed, the variations in several calibration parameters can be analyzed by comparing the base case model with the measure application scenarios. Depending on the measure, the results show changes of up to 6.4% in maximum speed values, and reductions of nearly 15% in acceleration and braking levels, especially when eco-driving is applied.

The role of rehabilitation of building in the urban integration, social cohesion and environmental responsibility

In the framework ofthe National Research Plan2008-2011, our research poses estrategy for the design and evaluation of plans and programmes of urban integrated regeneration. The objective is to develop a study on the role of rehabilitation of buildings in concepts like urban integration, social cohesion and environmental responsibility. The research proposes a methodological tool for evaluating urban regeneration processes from a holistic perspective that can serve as a guide for governments and technical teams to address intervention in consolidated urban areas with physical and socio-economic problems. The development of the tool has inevitably led to delve into different areas where you can intervene but has not lost sight of the complex interplay of factors involved in the process.It is an open source tool to visualize Urban Integrated Rehabilitation processes.

New agri-environmental policies for the sustainability of mountainous olive farming. Evidences from Andalusia, Spain

La sostenibilidad de los sistemas olivareros situados en zonas de pendiente y montaña (SMOPS) en Andalucía se encuentra actualmente amenazada por las elevadas tasas de abandono que afectan a estos sistemas productivos. Así, la consumación de este proceso de abandono, no sólo pondría en peligro a las propias explotaciones, sino a todo el conjunto de bienes y servicios no productivos y al patrimonio cultural generado por este sistema productivo. En este contexto, la búsqueda de alternativas políticas enfocadas a revertir este proceso se erige como una necesidad categórica en aras de garantizar en el largo plazo la sostenibilidad de los olivares de montaña. Esta tesis pretende hacer frente a esta necesidad a través de la construcción de un marco político alternativo para los SMOPS, que permita la integración simultánea de todas las dimensiones que pueden influir en su desarrollo; esto es: el marco político actual, principalmente determinado por la Política Agraria Común (PAC) de la Unión Europea (UE); las preferencias de la sociedad hacia la oferta de bienes y servicios públicos generados por los SMOPS; y las preferencias y voluntad de innovación hacia nuevos manejos y sistemas de gestión de los agricultores y propietarios de las explotaciones. Para ello, se emplea una metodología de investigación mixta, que abarca la realización de cuatro encuestas (personales y online) llevadas a cabo a los agentes o grupos de interés involucrados directa o indirectamente en la gestión de los SMOPS –ciudadanos, agricultores y propietarios y expertos-; una profunda revisión de las herramientas de política agroambiental actuales y posibles alternativas a las mismas; y el desarrollo de nuevas estrategias metodológicas para dotar de mayor precisión y fiabilidad las estimaciones obtenidas a partir del Método del Experimento de Elección (MEE) en el campo de la valoración medioambiental. En general, los resultados muestran que una estrategia de política agroambiental basada en la combinación de los Contratos Territoriales de Zona Rural (CTZR) y el manejo ecológico supondría una mejora en la sostenibilidad de los sistemas olivareros de montaña andaluces, que, al mismo tiempo, propiciaría una mejor consideración de las necesidades y demandas de los agentes implicados en su gestión. Asimismo, los hallazgos obtenidos en esta investigación demandan un cambio de paradigma en los actuales pagos agroambientales, que han de pasar de una estrategia basada en la implementación de acciones, a otra enfocada al logro de objetivos, la cual, en el caso del olivar, se podría centrar en el aumento del secuestro de carbono en el suelo. Desde un punto de vista metodológico, los resultados han contribuido notablemente a mejorar la fiabilidad y precisión de las conclusiones estimadas a partir del MEE, mediante el diseño de un novedoso proceso iterativo para detectar posibles comportamientos inconsistentes por parte de los entrevistados con respecto a su máxima Disposición al Pago (DAP) para lograr la situación considerada como “óptima” en los olivares ecológicos de montaña andaluces. En líneas generales, el actual marco institucional favorece la puesta en práctica de la mayoría de las estrategias propuestas en esta tesis; sin embargo son necesarios mayores esfuerzos para reconducir los actuales Pagos Agroambientales y Climáticos de la PAC, hacia una estrategia de política agroambiental adaptada a las necesidades y requisitos del territorio en el que se aplica, enfocada al logro de objetivos y que sea capaz de integrar y coordinar al conjunto de agentes y grupos de interés involucrados -directa o indirectamente- en la gestión de los olivares de montaña. En este contexto, se espera que la puesta en práctica de nuevas estructuras y acuerdos de gobernanza territorial juegue un importante papel en el desarrollo de una política agroambiental realmente adaptada a las necesidades de los sistemas olivareros de montaña andaluces. ABSTRACT The long-term sustainability of Andalusian sloping and mountainous olive production systems (SMOPS) is currently threatened by the high abandonment rates that affect these production systems. The effective occurrence of this abandonment process is indeed menacing not only farms themselves, but also the wide array of public goods and services provided by SMOPS and the cultural heritage held by this production system. The search of policy alternatives aimed at tackling this process is thus a central necessity. This thesis aims to undertake this necessity by building an alternative policy framework for SMOPS that simultaneously integrates the several dimensions that are susceptible to influence it, namely: the current policy framework, mainly determined by the European Union’s (EU) Common Agricultural Policy (CAP); the social preferences toward the supply of SMOPS’ public goods and services; and farmers’ preferences and willingness to innovate toward new management practices in their farms. For this purpose, we put into practice a mixed-method strategy that combines four face-to-face and online surveys carried out with SMOPS’ stakeholders -including citizens, farmers and experts-; in-depth analysis of current and alternative agrienvironmental policy (AEP) instruments; and the development of novel methodological approaches to advance toward more reliable Discrete Choice Experiment’s (DCE) outcomes in the field of environmental valuation. Overall, results show that a policy strategy based on the combination of Territorial Management Contracts (TMC) and organic management would further enhance Andalusian SMOPS’ sustainability by simultaneously taking into account stakeholders’ demands and needs. Findings also call for paradigm shift of the current action-oriented design of Agri-Environmental-Climate Schemes (AECS), toward a result-based approach, that in the case of olive orchards should particularly be focused on enhancing soil carbon sequestration. From a methodological perspective, results have contributed to improve the accuracy and feasibility of DCE outcomes by designing a novel and iterative procedure focused in ascertaining respondents’ inconsistent behaviour with respect to their stated maximum WTP for the attainment of an ideal situation to be achieved in organic Andalusian SMOPS. Generally, the present institutional framework favours the implementation of the main policy strategies proposed in this thesis, albeit further efforts are required to better conduct current CAP’s agri-environmental instruments toward a territorially targeted result-oriented strategy capable to integrate and coordinate the whole set of stakeholders involved in the management of SMOPS. In this regard, alternative governance structures and arrangements are expected to play a major role on the process of tackling SMOPS’ agri-environmental policy challenge.

Towards Mapping Experience Design for the Internet of Things.

We are witnessing a fundamental transformation in how Internet of Things (IoT) is having an impact on the experience users have with data-driven devices, smart appliances, and connected products. The experience of any place is commonly defined as the result of a series of user engagements with a surrounding place in order to carry out daily activities (Golledge, 2002). Knowing about users? experiences becomes vital to the process of designing a map. In the near future, a user will be able to interact directly with any IoT device placed in his surrounding place and very little is known on what kinds of interactions and experiences a map might offer (Roth, 2015). The main challenge is to develop an experience design process to devise maps capable of supporting different user experience dimensions such as cognitive, sensory-physical, affective, and social (Tussyadiah and Zach, 2012). For example, in a smart city of the future, the IoT devices allowing a multimodal interaction with a map could help tourists in the assimilation of their knowledge about points of interest (cognitive experience), their association of sounds and smells to these places (sensory-physical experience), their emotional connection to them (affective experience) and their relationships with other nearby tourists (social experience). This paper aims to describe a conceptual framework for developing a Mapping Experience Design (MXD) process for building maps for smart connected places of the future. Our MXD process is focussed on the cognitive dimension of an experience in which a person perceives a place as a "living entity" that uses and feeds through his experiences. We want to help people to undergo a meaningful experience of a place through mapping what is being communicated during their interactions with the IoT devices situated in this place. Our purpose is to understand how maps can support a person?s experience in making better decisions in real-time.

Synaptic vesicle endocytosis mediates the entry of tetanus neurotoxin into hippocampal neurons

Tetanus neurotoxin causes the spastic paralysis of tetanus by blocking neurotransmitter release at inhibitory synapses of the spinal cord. This is due to the penetration of the toxin inside the neuronal cytosol where it cleaves specifically VAMP/synaptobrevin, an essential component of the neuroexocytosis apparatus. Here we show that tetanus neurotoxin is internalized inside the lumen of small synaptic vesicles following the process of vesicle reuptake. Vesicle acidification is essential for the toxin translocation in the cytosol, which results in the proteolytic cleavage of VAMP/synaptobrevin and block of exocytosis.

Identification of two residues in MCM5 critical for the assembly of MCM complexes and Stat1-mediated transcription activation in response to IFN-γ

In response to IFN-γ, the latent cytoplasmic Stat1 (signal transducer and activator of transcription) proteins translocate into the nucleus and activate transcription. We showed previously that Stat1 recruits a group of nuclear proteins, among them MCM5 (minichromosome maintenance) and MCM3, for transcription activation. MCM5 directly interacts with the transcription activation domain (TAD) of Stat1 and enhances Stat1-mediated transcription activation. In this report, we identified two specific residues (R732, K734) in MCM5 that are required for the direct interaction between Stat1 and MCM5 both in vitro and in vivo. MCM5 containing mutations of R732/K734 did not enhance Stat1-mediated transcription activation in response to IFN-γ. In addition, it also failed to form complexes with other MCM proteins in vivo, suggesting that these two residues may be important for an interaction domain in MCM5. Furthermore, MCM5 bearing mutations in its ATPase and helicase domains did not enhance Stat1 activity. In vitro binding assays indicate that MCM3 does not interact directly with Stat1, suggesting that the presence of MCM3 in the group of Stat1TAD-interacting proteins is due to the association of MCM3 with MCM5. Finally, gel filtration analyses of nuclear extracts from INF-γ-treated cells demonstrate that there is a MCM5/3 subcomplex coeluting with Stat1. Together, these results strongly suggest that Stat1 recruits a MCM5/3 subcomplex through direct interaction with MCM5 in the process of IFN-γ-induced gene activation.

Bacteria are different: Observations, interpretations, speculations, and opinions about the mechanisms of adaptive evolution in prokaryotes

To some extent, the genetic theory of adaptive evolution in bacteria is a simple extension of that developed for sexually reproducing eukaryotes. In other, fundamental ways, the process of adaptive evolution in bacteria is quantitatively and qualitatively different from that of organisms for which recombination is an integral part of the reproduction process. In this speculative and opinionated discussion, we explore these differences. In particular, we consider (i) how, as a consequence of the low rates of recombination, “ordinary” chromosomal gene evolution in bacteria is different from that in organisms where recombination is frequent and (ii) the fundamental role of the horizontal transmission of genes and accessory genetic elements as sources of variation in bacteria. We conclude with speculations about the evolution of accessory elements and their role in the adaptive evolution of bacteria.

Fused polycationic peptide mediates delivery of diphtheria toxin A chain to the cytosol in the presence of anthrax protective antigen.

The lethal factor (LF) and edema factor (EF) of anthrax toxin bind by means of their amino-terminal domains to protective antigen (PA) on the surface of toxin-sensitive cells and are translocated to the cytosol, where they act on intracellular targets. Genetically fusing the amino-terminal domain of LF (LFN; residues 1-255) to certain heterologous proteins has been shown to potentiate these proteins for PA-dependent delivery to the cytosol. We report here that short tracts of lysine, arginine, or histidine residues can also potentiate a protein for such PA-dependent delivery. Fusion of these polycationic tracts to the amino terminus of the enzymic A chain of diphtheria toxin (DTA; residues 1-193) enabled it to be translocated to the cytosol by PA and inhibit protein synthesis. The efficiency of translocation was dependent on tract length: (LFN > Lys8 > Lys6 > Lys3). Lys6 was approximately 100-fold more active than Arg6 or His6, whereas Glu6 and (SerSerGly)2 were inactive. Arg6DTA was partially degraded in cell culture, which may explain its low activity relative to that of Lys6DTA. The polycationic tracts may bind to anionic sites at the cell surface (possibly on PA), allowing the fusion proteins to be coendocytosed with PA and delivered to the endosome, where translocation to the cytosol occurs. Excess free LFN blocked the action of LFNDTA, but not of Lys6DTA. This implies that binding to the LF/EF site is not an obligatory step in translocation and suggests that the polycationic tag binds to a different site. Besides elucidating the process of translocation in anthrax toxin, these findings may aid in developing systems to deliver heterologous proteins and peptides to the cytoplasm of mammalian cells.

Surface protein phosphorylation by ecto-protein kinase is required for the maintenance of hippocampal long-term potentiation.

During the induction of long-term potentiation (LTP) in hippocampal slices adenosine triphosphate (ATP) is secreted into the synaptic cleft, and a 48 kDa/50 kDa protein duplex becomes phosphorylated by extracellular ATP. All the criteria required as evidence that these two proteins serve as principal substrates of ecto-protein kinase activity on the surface of hippocampal pyramidal neurons have been fulfilled. This phosphorylation activity was detected on the surface of pyramidal neurons assayed after synaptogenesis, but not in immature neurons nor in glial cells. Addition to the extracellular medium of a monoclonal antibody termed mAb 1.9, directed to the catalytic domain of protein kinase C (PKC), inhibited selectively this surface protein phosphorylation activity and blocked the stabilization of LTP induced by high frequency stimulation (HFS) in hippocampal slices. This antibody did not interfere with routine synaptic transmission nor prevent the initial enhancement of synaptic responses observed during the 1-5 min period immediately after the application of HFS (the induction phase of LTP). However, the initial increase in the slope of excitatory postsynaptic potentials, as well as the elevated amplitude of the population spike induced by HFS, both declined gradually and returned to prestimulus values within 30-40 min after HFS was applied in the presence of mAb 1.9. A control antibody that binds to PKC but does not inhibit its activity had no effect on LTP. The selective inhibitory effects observed with mAb 1.9 provide the first direct evidence of a causal role for ecto-PK in the maintenance of stable LTP, an event implicated in the process of learning and the formation of memory in the brain.

Correlation of individual papilloma latency time with DNA adducts, 8-hydroxy-2'-deoxyguanosine, and the rate of DNA synthesis in the epidermis of mice treated with 7,12-dimethylbenz[alpha]anthracene.

The question was addressed whether the risk of cancer of an individual in a heterogeneous population can be predicted on the basis of measurable biochemical and biological variables postulated to be associated with the process of chemical carcinogenesis. Using the skin tumor model with outbred male NMRI mice, the latency time for the appearance of a papilloma was used as an indicator of the individual cancer risk. Starting at 8 weeks of age, a group of 29 mice was treated twice weekly with 20 nmol of 7,12-dimethylbenz[alpha]anthracene (DMBA) applied to back skin. The individual papilloma latency time ranged from 13.5 to 25 weeks of treatment. Two weeks after the appearance of the first papilloma in each mouse, an osmotic minipump delivering 5-bromo-2'-deoxyuridine was s.c. implanted and the mouse was killed 24 hr later. Levels of DMBA-DNA adducts, of 8-hydroxy-2'-deoxyguanosine, and various measures of the kinetics of cell division were determined in the epidermis of the treated skin area. The levels of 8-hydroxy-2'-deoxyguanosine and the fraction of cells in DNA replication (labeling index for the incorporation of 5-bromo-2'-deoxyuridine) were significantly higher in those mice that showed short latency times. On the other hand, the levels of DMBA-DNA adducts were lowest in animals with short latency times. The latter finding was rather unexpected but can be explained as a consequence of the inverse correlation seen for the labeling index: with each round of cell division, the adduct concentration is reduced to 50% because the new DNA strand is free of DMBA adducts until the next treatment. Under the conditions of this bioassay, therefore, oxygen radical-related genotoxicity and the rate of cell division, rather than levels of carcinogen-DNA adducts, were found to be of predictive value as indicators of an individual cancer risk.

Stability and sensitivity analysis of Be-CoDiS, an epidemiological model to predict the spread of human diseases between countries. Validation with data from the 2014-16 West African Ebola Virus Disease epidemic.

Ebola virus disease is a lethal human and primate disease that requires a particular attention from the international health authorities due to important recent outbreaks in some Western African countries and isolated cases in European and North-America continents. Regarding the emergency of this situation, various decision tools, such as mathematical models, were developed to assist the authorities to focus their efforts in important factors to eradicate Ebola. In a previous work, we have proposed an original deterministic spatial-temporal model, called Be-CoDiS (Between-Countries Disease Spread), to study the evolution of human diseases within and between countries by taking into consideration the movement of people between geographical areas. This model was validated by considering numerical experiments regarding the 2014-16 West African Ebola Virus Disease epidemic. In this article, we propose to perform a stability analysis of Be-CoDiS. Our first objective is to study the equilibrium states of simplified versions of this model, limited to the cases of one an two countries, and to determine their basic reproduction ratios. Then, in order to give some recommendations for the allocation of resources used to control the disease, we perform a sensitivity analysis of those basic reproduction ratios regarding the model parameters. Finally, we validate the obtained results by considering numerical experiments based on data from the 2014-16 West African Ebola Virus Disease epidemic.